Methods of treating cancers

Abstract

Provided are methods, and reagents and kits for performing such methods, directed to treating subjects with cancer, including subjects with hematological cancers and subjects with solid tumors, including subjects with relapsed and / or refractory hematological cancers, subjects with metastatic, recurrent, relapsed, unresectable, and / or advanced tumors. Provided are methods of treating cancer comprising administering a decoy-resistant interleukin 18 (DR-18), including e.g., using combination therapies comprising administering a DR-18 and a multi-specific therapeutic. Methods may also include selecting a patient, and administering a DR-18 to result in enhanced cytotoxic activity of immune cells in the patient to treat the subject for cancer. Also provided are methods for administration of DR-18, to prime immune cells present in the subject, and administration of one or more agents that activate the primed immune cells, resulting in enhanced cytotoxic activity, thereby treating the cancer.

Description

[0001] Attorney Docket No.: S199207 1060WO (00093)

[0002] ST-020-W01

[0003] Methods of Treating Cancers

[0004] Cross-Reference to Related Applications

[0005] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 687,114, filed on August 26, 2024. The entire contents of the foregoing application are hereby incorporated herein by reference.

[0006] Sequence Listing

[0007] The instant application contains a Sequence Listing which has been filed electronically in extensible Markup Language (XML) format and is hereby incorporated by reference in its entirety. Said XML copy, created on August 26, 2025, is named S199207_1060WO_SL.xml and is 84,215 bytes in size.

[0008] Introduction

[0009] The effectiveness of treatments for treatment-naive hematological malignancies varies depending upon the specific type and stage of the cancer. Conventional approaches, such as chemotherapy and radiation, have demonstrated efficacy in inducing remission in a significant portion of subjects; however, the associated adverse effects and potential for relapse necessitate the exploration of alternative and adjunctive therapies. Targeted therapies, including monoclonal antibodies and small molecule inhibitors, represent advancements in treatment strategies by leveraging molecular pathways implicated in hematological malignancies. Additionally, recent developments in immunotherapy, such as chimeric antigen receptor T-cell (CAR-T) therapy and immune checkpoint inhibitor (ICI) therapy, provide promising outcomes by enhancing the body's immune response to malignancies. However, CAR-T therapy has certain practical obstacles in that each therapy must be produced individually for each patient and results from trials of ICI therapies in hematological malignancies have been underwhelming, at least in part due to resistance mechanisms in ICI monotherapy and toxicity in multiple ICI combination therapy (Tang et al. (2023) Signal Transduction and Targeted Therapy 8(1 ):306). Resistance to monotherapy, whether cancer-targeting antibodies or I Cis, and combination therapy toxicity present significant opposing barriers to achieving long-term remission and survival.

[0010] About 35,780 new cases of multiple myeloma are expected to be diagnosed, and an estimated 12,540 deaths are expected to occur from multiple myeloma in 2024 (American Attorney Docket No.: S199207 1060WO (00093)

[0011] ST-020-W01

[0012] Cancer Society (ACS), Key Statistics for Multiple Myeloma, 2024). Non-Hodgkin lymphoma (NHL) is one of the most common cancers in the United States, accounting for about 4% of all cancers. It is estimated that about 80,620 people (including adults and children) will be diagnosed with NHL and about 20,140 people will die from this cancer in 2024 (ACS. Key Statistics for Non-Hodgkin Lymphoma, 2024). It is estimated that, in 2024, acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia (CML) will collectively account for about 57,330 new cases of cancer (with AML and CLL each accounting for about one third) and about 18,270 deaths (ACS, Key Statistics for Leukemia, 2024). Furthermore, incidence and mortality are not necessarily evenly distributed. For example, while most cases of ALL occur in children, most deaths from ALL (about 4 out of 5) occur in adults.

[0013] Therapies for newly diagnosed hematological malignancies, including chemotherapy and radiation, demonstrate efficacy in achieving remission. For instance, the 5-year relative survival rate for individuals with Hodgkin lymphoma treated with these approaches is 88.3%, and for those with non-Hodgkin lymphoma, the rate is 73.2% (National Cancer Institute (NCI), SEER Cancer Statistics, 2022). Leukemia, specifically ALL, has a 5-year relative survival rate of approximately 69.9% when treated with standard chemotherapy protocols, whereas the 5-year survival rate for AML is near 30% (NCI, SEER Cancer Statistics, 2022). Despite initial positive outcomes, a significant proportion of subjects experience relapse. For example, in AML, relapse occurs in about 40-50% of patients who achieve remission after first-line treatment (ACS, Acute Myeloid Leukemia, 2022).

[0014] Moreover, relapsed disease is often refractory to the prior line of therapy. The recurrence of refractory conditions, where cancer becomes resistant to treatment, poses a substantial challenge. For example, almost all patients with multiple myeloma eventually relapse and relapsed / refractory multiple myeloma has limited therapeutic options, leading to median survival rates of approximately only 5-9 months (Kumar et al. (2012) Leukemia 26(1 ): 149— 157). The necessity to address relapsed and refractory cancers, including relapsed / refractory hematological cancers, remains a concern in the development of new therapeutic strategies.

[0015] For example, colorectal cancer (CRC), also known as bowel cancer or colon cancer, the fourth most common cancer diagnosis and second leading cause of cancer-related death in the U.S., has an approximately 50% rate of development of distant recurrence and metastatic CRC and, with limited exceptions, is incurable and associated with a median overall survival rate of less than 2 years (AJMC Perspectives, July 2024 “Strategies for the Management of Relapsed / Refractory Metastatic Colorectal Cancer: From Landmark Trials to Precision Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0016] Medicine”). Even for microsatellite instability-high (MSI-H) metastatic CRC, where MSI-H is a major predictive biomarker for the efficacy of ICI therapy, nearly 10% of initially responding patients develop resistance to ICI therapy (Loupakis et al., (2020) Oncologist, 25(6):481-487).

[0017] Developed (or alternatively referred to as “secondary”) resistance is not limited to ICI therapy. Bispecific antibodies, including bispecific T cell engagers (BiTEs), although effective (see e.g., Curran & Stock (2019) Blood. 133(16): 1715— 1719) are also subject to resistance mechanisms that can decrease the effectiveness of the originally administered therapeutic leading to relapse / refractor disease. For example, resistance mechanisms, including lineage switching and target antigen downregulation, have been observed in ALL patients treated with blinatumomab, an anti-CD19 / CD3 BiTE (see e.g., Haddox et al. (2017) Blood Cancer J. 7(9):e607 and Mejstrikova, et al. (2017) Blood Cancer J. 7(12):659).

[0018] Summary

[0019] The present invention provides a method of treating a subject having a cancer, such as a hematological cancer or a solid tumor, the method comprising administering a decoy-resistant interleukin 18 (DR-18) to the subject, thereby treating the subject.

[0020] The present invention also provides a method of priming immune cells of a subject having a cancer, such as a hematological cancer or a solid tumor, the method comprising administering a decoy- resista nt interleukin 18 (DR-18) to the subject, thereby priming the immune cells of the subject. In certain aspects, this method also includes administering to the subject one or more agents that activate the primed immune cells.

[0021] The present invention also provides a method of enhancing cytotoxicity of immune cells in a subject having a cancer, such as a hematological cancer of a solid tumor, the method comprising administering a decoy-resistant interleukin 18 (DR-18) to the subject, thereby enhancing cytotoxicity of immune cells against the cancer.

[0022] In respect to the afore-mentioned methods, in some aspects, the subject has a hematological cancer that is relapsed and / or refractory to at least one standard of care (SOC) therapy, and / or the subject is deficient in the production of anti-drug antibodies. In particular aspects of the disclosed methods, the hematological cancer is a lymphoma, a leukemia, or multiple myeloma. Also contemplated herein, the hematological cancer is a B cell cancer, including for example, a B cell cancer that expresses an antigen selected from CD19, CD20, BCMA, CD123, CD38, CD33, FcRH5, and GPRC5D.

[0023] In other aspects, the disclosed methods include measuring, in a sample obtained from the subject, an amount of B cells present in the sample and / or an amount of plasma cells Attorney Docket No.: S199207 1060WO (00093)

[0024] ST-020-W01 present in the sample. This measurement of B cells and / or plasma cells can be performed to assist in patient selection and / or to monitor ongoing treatment. For example, the measuring can be performed prior to the administering, optionally wherein the subject is selected for administration of the DR-18 when the measured amount of B cells and / or plasma cells is less than an amount expected in a corresponding healthy subject. In particular aspects, a patient is selected for treatment when the measured amount of B cells is about 10% or less of the total lymphocyte population in the subject, optionally wherein the measured amount of B cells is 200 or less cells per microliter. As another example, the measuring can be performed after the administering for monitoring of the treatment.

[0025] In all of the disclosed methods and kits, it is contemplated that the DR-18 can have less than 100% sequence identity and at least 85% sequence identity to wild-type human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXXAVTISV KXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEGYFLAXEKERD LFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO:64), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and / or the X at pos. 155 is N, K or H.

[0026] For example, the DR-18 can comprise an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62 and 63. In particular aspects, the DR-18 can comprise an amino acid sequence set forth as SEQ ID NO: 20. As another example, the DR18 can consist of, or consist essentially of, an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62 and 63. For instance, the DR- 18 can consist of, or consist essentially of, an amino acid sequence set forth as SEQ ID NO: 20.

[0027] The present invention contemplates that the DR-18 can be administered in a singular dose or a plurality of doses. In either instance, a representative dose is an amount from 20 pg / kg to 1200 pg / kg, or an amount from 30 pg / kg to 1200 pg / kg, optionally from 60 pg / kg to 240 pg / kg. If multiple doses are uses, each dose is separated by at least five days and no more than nine days. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0028] In still other aspects of the invention, the method further comprises administering to the subject a multi-specific therapeutic, wherein the multi-specific therapeutic binds to an antigen expressed on cells of the hematological cancer. In particular aspects of the invention, the multispecific therapeutic binds to the antigen expressed on cells of the hematological cancer and to an antigen expressed on immune cells in the subject. In other particular aspects, the multispecific therapeutic is a bispecific therapeutic.

[0029] Administration of the DR-18 and the multi-specific therapeutic can be done concurrently or sequentially in either order. In some aspects, the multi-specific therapeutic is administered after the first dose of the DR-18, and optionally at least 24 hours after, optionally at least one week after.

[0030] Any multispecific antibody that comprises a component of binding to a cancer antigen is contemplated by the invention. In particular aspects the cancer antigen is a hematological cancer antigen, and in other aspects, the cancer antigen is a solid tumor antigen. The one or more binding specificities of the multispecific antigen can include a second cancer antigen and / or a immune cell antigen.

[0031] In particular aspects of the disclosed combination therapies that include DR-18 and a mutli-specific antibody, the multi-specific therapeutic is selected from blinatumomab, mosunetuzumab, epcoritamab, teclistamab, flotetuzumab, talquetamab, and AMG 330. In specific aspects of the invention, the multi-specific antibody included in the combination therapy is blinatumomab.

[0032] Subjects treatable by the disclosed methods include naive, relapsed, and refractory patients.

[0033] In the disclosed methods for DR-18 therapies for hematological cancer, assessing performance of the method, in some cases treating a subject, can be done using any one of numerous routine and well-understood techniques and / or clinical measures. For example, in some aspects, the treating results in a reduction in the number of cancerous cells in the subject, optionally as measured by a complete blood count, peripheral blood smear, bone marrow biopsy, flow cytometry, or a molecular diagnostic assay. In other aspects, the treating results in increased cytotoxic killing of hematological cancer cells. In still other aspects, the treating results in amelioration of at least one symptom of the hematological cancer, for example, a minimal / measurable residual disease (MRD) negativity, prevention of MRD positivity or sustained MRD negativity, a partial therapeutic response, and / or a complete therapeutic response. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0034] In the disclosed methods, assessing performance of the method is measured as an increase in life expectancy of the subject. For example, the increase in life expectancy of the subject is an at least 10%, at least 15%, or at least 20% increase in life expectancy of the subject. In some particular aspects, the life expectancy of the subject is increased as compared to the subject’s life expectancy if treated according to the standard of care therapy for the cancer, and in some aspects where no standard of care therapy for the hematological cancer is available that prolongs survival.

[0035] The instant application also provides a method and combination therapy of treating a subject having a cancer, the method comprising administering to the subject a decoy-resistant interleukin 18 (DR-18) and a multi-specific therapeutic, wherein the multi-specific therapeutic binds to an antigen expressed on cells of the cancer.

[0036] In particular aspects of the disclosed combination therapy, the multi-specific therapeutic binds to an antigen expressed on cells of cancer cells. In other particular aspects, the multispecific therapeutic is a bispecific therapeutic. In particular aspects of the invention, the multispecific therapeutic binds to the antigen expressed on cells of the cancer and to an antigen expressed on immune cells in the subject. In other particular aspects, the multi-specific antibody is a T-cell engager. In other particular aspects, the antigen expressed on immune cells is a CD3 antigen.

[0037] The disclosed combination therapies comprising a DR-18 and a multispecific therapeutic are useful for treatment of both hematologic malignancies and solid tumors.

[0038] In particular aspects, the antigen expressed on cells of the cancer is selected from CD19, EpCAM, CD20, GP100, BCMA, EGFR, cMET, HER2, HER3, CLDN18.2, and TRP-1 ; for example an antigen selected from CD19, EpCAM, CD20, GP100, and BCMA; or for example an antigen expressed on cells of the cancer is selected from CD19, CD20, BCMA, CLDN18.2, and TRP-1.

[0039] In particular aspects, the disclosed combination therapies include administering a multispecific therapeutic that binds to CD3 x gp100, CD3 x DLL3, CD3 x PRAME, CD3 x STEAP1 , CD3 x MUC16, CD3 x DLL3, CD3 x PIWIL1, CD3s x DLL3 x albumin, CD3 x HER2, CD3 x CLDN18.2, CD3 x CLDN6, CD3 x HLA x MAGEA4, CD3 x PSMA, CD3 x CEA, CD3 x EGFR, CD3 x PRAME, CD3 x EpCAM, CD3 x EGFR, CD3 x PSMA, CD3 x GPC3, CD3 x CDH3 x MSLN, CD3 x CDH17, CD3 x TRP-1, CD3 x NCR3LG1 , CD3 x 5T4, CD3 x CLDN6, CD3 x CLDN6, CD3 x EGFR, CD3 x KLK2, CD3 x ENPP3, CD3 x CALR, CD3 x CEACAM5, CD3 x ROR1 , CD3 x EGFRvlll, CD3 x CD3E x HER2, CD3 x 4-1 BB x DLL3, CD3 x CLDN6, CD3 x ENPP3, CD3 x MSLN, CD3 x PSMA, CD3 x MSLN, CD3 x CD20, CD3 x GPRC5D, CD3 x Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0040] BCMA, CD3 x CD19, CD3 x BCMA, CD3 x CD20, CD3 x CD20, CD3 x CD20, CD3 x BCMA, CD3 x CD19 x CD3D, CD3 x CD123, CD3 x FCRL5, CD3 x BCMA, CD3 x GPRC5D, CD3 x CD123, CD3 x FLT3, CD3 x CD19, CD3 x CD38, CD3 x BCMA x CD38, CD3 x CD22, CD3 x BCMA x GPRC5D, CD3 x CD20 x CD79B, CD3 x CD123, CD3 x PD-1, CD3 x BCMA, CD3 x CD19 x CD2, CD3 x CD38, CD3 x BCMA, CD3 x EpCAM, CD3 x MUC16, CD3 x DLL3, CD3 x GPC3, CD3 x CD19 x CD20 x HER2, CD3 x HER2, CD3 x TAA, CD3 x CD33, CD3 x CD276, CD3 x EpCAM, CD3 x BCMA x GPRC5D, CD3 x CD19 x CD20, CD3 x BCMA, CD3 x CD20, CD3 x CD20, CD3 x BCMA x GPRC5D, CD3 x GPRC5D, CD3 x CD19 x CD28, CD3 x BCMA x GPRC5D, or CD3 x CD276.

[0041] In particular aspects, the disclosed combination therapies include administering a multispecific therapeutic that is selected from blinatumomab, amivantamab, catumaxomab, mosunetuzumab, epcoritamab, tebentafusp, teclistamab, and zenocutuzumab. In particular aspects, the multi-specific therapeutic is selected from blinatumomab, catumaxomab, mosunetuzumab, epcoritamab, tebentafusp, and teclistamab, and more particularly, blinatumomab.

[0042] In other particular aspects, the disclosed combination therapies include administering a multi-specific therapeutic that is selected from givastomig, gresonitamab, spevatamig, and tixentamig, more particularly gresonitamab or tixentamig.

[0043] In the disclosed methods for DR-18 combination therapies for cancer, assessing performance of the method, in some cases treating a subject, can be done using any one of numerous routine and well-understood techniques and / or clinical measures. For example, in some aspects, the treating results in a reduction in the amount of cancerous cells in the subject, optionally as measured by a complete blood count, peripheral blood smear, bone marrow biopsy, flow cytometry, or a molecular diagnostic assay. In other aspects, the treating results in increased cytotoxic killing of hematological cancer cells. In still other aspects, the treating results in amelioration of at least one symptom of the hematological cancer, for example, a minimal / measurable residual disease (MRD) negativity, prevention of MRD positivity or sustained MRD negativity, a partial therapeutic response, and / or a complete therapeutic response.

[0044] In the disclosed combination therapies, assessing performance of the method can be measured as an increase in life expectancy of the subject. For example, the increase in life expectancy of the subject is an at least 10%, at least 15%, or at least 20% increase in life expectancy of the subject. In some particular aspects, the life expectancy of the subject is increased as compared to the subject’s life expectancy if treated according to the standard of Attorney Docket No.: S199207 1060WO (00093)

[0045] ST-020-W01 care therapy for the cancer, and in some aspects where no standard of care therapy for the hematological cancer is available that prolongs survival.

[0046] Also provided are kits comprising the disclosed DR-18 therapy and combination therapies. For example, a kit provided herein includes DR-18 for the treatment of a hematological cancer. As another example, a kit provided herein includes a decoy-resistant interleukin 18 (DR-18) polypeptide in a first container, and a multi-specific antibody in a second container, wherein the multi-specific antibody binds to an antigen expressed on cells of the cancer. In various aspects, the disclosed kits can be formatted, for example, as a pre-filled syringe for subcutaneous administration, and can include instructional material providing a recommended dosing schedule for the DR-18 polypeptide, optionally in combination with a multispecific antibody as disclosed herein.

[0047] Brief Description of the Figures

[0048] FIG. 1 depicts inhibition of C1498 acute myeloid leukemia (AML) tumor growth in mice administered mouse decoy-resistant interleukin 18 (DR-18), ST-001. Negative control (“vehicle”) and standard of care (SOC) 5-Azacitidine / Venetoclax therapy (“Aza-VentoClax”) groups are included for comparison. Y-axis represents tumor size in cubic millimeters (mm3).

[0049] FIG. 2 depicts increased survival of C1498 AML tumor bearing mice administered mouse DR-18, ST-001 , including as a monotherapy or in combination with an azacytidine and venetoclax regimen.

[0050] FIG. 3 depicts decreased tumor growth inhibition of mouse DR-18, ST-001 , in mice subjected to CD8+ T cell depletion as compared to mice subjected to only CD4+ T cell depletion, NK cell depletion, or no immune cell depletion. The groups listed in order, from left to right, are ST-001 (i.e., no immune cell depletion), anti-NK (i.e., NK cell depleted), anti-CD4 (i.e., CD4 T cell depleted), anti-CD8 (i.e., CD8 T cell depleted), anti-CD4 / CD8 (i.e., CD4 T cell and CD8 T cell depleted).

[0051] FIG. 4 depicts enhanced hematological tumor growth inhibition, as well as improved treatment durability, in mice treated with combination ST-001 plus blinatumomab therapy, e.g., as compared to either ST-001 or blinatumomab monotherapy alone.

[0052] FIG. 5 provides a schematic showing the design for an in vivo B cell lymphoma rechallenge mouse study as described herein.

[0053] FIG. 6 depicts inhibition of A20 B cell lymphoma (BCL) tumor growth in mice administered mouse decoy-resistant interleukin 18 (DR-18), ST-001. Negative control (“vehicle”) and standard of care (SOC) CPX-351 (CPX, a liposomal formulation of cytarabine and Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 daunorubicin) groups are included for comparison. Y-axis represents tumor volume in cubic millimeters (mm3).

[0054] FIG. 7 depicts the body weights of the mice shown in FIG. 6 as measured during the course of therapy, showing that DR-18 ST-001 is well-tolerated in the model.

[0055] FIG. 8 depicts inhibition of A20 tumor growth in mice previously treated with DR-18 ST- 001 and re-challenged with implantation of new A20 tumors at 32 days after the start (and 18 days after cessation) of the initial ST-001 treatment. See FIG. 5 for associated schematic. The growth of tumors in control naive mice that were implanted with the same A20 cells utilized in the rechallenge cohort is provided for reference.

[0056] FIG. 9 depicts enhanced inhibition of MC38-huCLDN18.2 colon cancer tumor growth in mice administered combination therapy of mouse decoy-resistant interleukin 18 (DR-18), ST- 001 plus anti-CD3-CLDN18.2 bispecific T cell engager, gresonitamab. Negative control (“vehicle”) and corresponding monotherapy groups at varied dosages are included for comparison. Y-axis represents tumor volume in cubic millimeters (mm3).

[0057] FIG. 10 depicts increased survival of MC38-huCLDN18.2 colon cancer tumor bearing mice administered combination therapy of mouse DR-18, ST-001 plus anti-CD3-CLDN18.2 bispecific T cell engager, gresonitamab, e.g., as compared to mice administered corresponding monotherapies.

[0058] FIG. 11 depicts the body weights of the mice shown in FIG. 10 as measured during the course of therapy, showing that the combination therapy of DR-18 ST-001 plus gresonitamab is well-tolerated in the model.

[0059] FIG. 12 depicts increased survival of B16-F10 melanoma bearing mice administered combination therapy of mouse DR-18, ST-001 plus anti-CD3-TRP1 bispecific T cell engager, e.g., as compared to mice administered corresponding monotherapies.

[0060] Definitions

[0061] The terms “subject,” “individual,” or “patient” are often used interchangeably herein. A “subject” comprises a biological entity containing expressed genetic materials. In some embodiments, the subject comprises an animal, mammal, primate, or human. In some embodiments, the subject is diagnosed with cancer. In some embodiments, the subject has a solid tumor. In some embodiments, the subject has a liquid tumor.

[0062] The term “subject in need thereof” as used herein refers to a subject diagnosed with or suspected of having cancer as defined herein. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0063] As used herein, the terms “treat,” “treated,” “treating,” “treatment,” and the like are used in reference to an intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to prevention or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with prevention or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or a condition, delaying or eliminating the onset of symptoms of a disease or a condition, slowing, halting, or reversing the progression of a disease or a condition, or any combination thereof. For prophylactic benefit, a subject at risk of developing a particular disease or a condition, or to a subject reporting one or more of the physiological symptoms of a disease or a condition may undergo treatment.

[0064] The term “prevent”, “preventing”, “prevention” and their grammatical equivalents as used herein, means avoiding or delaying the onset of symptoms associated with a disease or condition in a subject that has not developed such symptoms at the time the administering of an agent or compound commences.

[0065] The term “therapeutic effect” refers to some extent of relief of one or more of the symptoms of a disorder (e.g., a neoplasia, tumor, or infection by an infectious agent or an autoimmune disease) or its associated pathology. “Therapeutically effective amount” as used herein refers to an amount of an agent which is effective, upon single or multiple dose administration to the cell or subject, in prolonging the survivability of the patient with such a disorder, reducing one or more signs or symptoms of the disorder, preventing or delaying, and the like beyond that expected in the absence of such treatment. “Therapeutically effective amount” is intended to qualify the amount required to achieve a therapeutic effect. A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the “therapeutically effective amount” (e.g., ED50) of the pharmaceutical composition required.

[0066] The terms “neoplasia” and “cancer” refers to any disease that is caused by or results in inappropriately high levels of cell division, inappropriately low levels of apoptosis, or both. Merkel cell carcinoma is one non-limiting example of a cancer. The terms “cancer” or “tumor” or “hyperproliferative disorder” refer to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Cancer cells are Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 often in the form of a tumor, but such cells can exist alone within an animal, or can be a nontumorigenic cancer cell, such as a leukemia cell.

[0067] The term “liquid tumor” refers to cancers occurring, or the presence of cancerous cells, in bodily fluids. Accordingly, liquid tumors are cancers that affect the blood, bone marrow, and / or lymphatic systems. Liquid tumors include hematological (or “heme” or blood) cancers, such as e.g., a leukemia, a lymphoma, a myelodysplastic syndrome, a myeloproliferative disorder, and myeloma. The most common types of hematological cancers include leukemia, lymphoma, and myeloma. By contrast, a “solid tumor” refers to a grouping of cancerous cells that forms a solid mass in a tissue of a subject, such as bone tissue, muscle tissue, epithelial tissue, organ tissue (e.g., liver, lung, stomach, colon, kidney, or brain tissue), and the like. Types of solid tumors include e.g., carcinomas, sarcomas, solid tumor lymphomas, melanomas, neuroendocrine tumors, germ cell tumors, gliomas, and the like.

[0068] The term “lymphoma” refers to cancer that originates from the lymphatic system and starts in lymphocytes. There are two main types of lymphoma called Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL). In HL lymphocytes can grow out of control, causing swollen lymph nodes and growths throughout the body. Classic HL (cHL) is the most common HL and includes subtypes such as Nodular sclerosis Hodgkin lymphoma (NSCHL), Mixed cellularity Hodgkin lymphoma (MCCHL), Lymphocyte-rich HL, and Lymphocyte-depleted HL. Aside from cHL, Nodular lymphocyte-predominant HL (NLPHL) accounts for about 5% of cases. NHL is a diverse group of blood cancers that includes all types of lymphoma except Hodgkin’s lymphomas. NHL can begin in B lymphocytes (also known as B cells) or T lymphocytes (also known as T cells). Types of B cell lymphoma (BCL) include but are not limited to large B-cell lymphoma (LBCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphomas (including e.g., extranodal marginal zone B-cell lymphoma, nodal marginal zone B-cell lymphoma, and splenic marginal zone B-cell lymphoma), and Burkitt lymphoma. Types of T cell lymphoma include but are not limited to lymphoblastic lymphoma / leukemia, cutaneous T-cell lymphomas, angioimmunoblastic T-cell lymphoma (AITL), enteropathy-associated intestinal T-cell lymphoma (EATL), and anaplastic large cell lymphoma (ALCL).

[0069] The term “leukemia” refers to cancer that originates in early forming blood cells, including white blood cells or other blood cell types. Types of leukemia include acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and chronic myelomonocytic leukemia (CMML). ALL (or Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 acute lymphoblastic leukemia) originates in lymphoid cells and there are two main types, B-cell ALL which originates in early forms of B lymphocytes and the less common T-cell ALL which originates in early forms of T lymphocytes. AML originates in the bone marrow in myeloid cells, there are many different subtypes, and AML is one of the most common leukemias in adults. CLL originates in lymphocytes in the bone marrow and accounts for about one-third of all leukemias with B-cell CLL (B-CLL) being the most common form. CML, also known as chronic myelogenous leukemia, originates in myeloid cells and results in abnormal production of granulocytes. CM ML is a rare myelodysplastic / myeloproliferative neoplasm and can lead to the development of AML.

[0070] “Multiple myeloma”, also referred to as “myeloma”, originates in plasma cells, is characterized by the abnormal increase of monoclonal immunoglobulins, is relatively uncommon, and has a variable prognosis affected by multiple factors.

[0071] An “agent” can include any type of molecule and includes, but is not limited to, an antibody, a peptide, a protein, a polynucleotide (e.g., an oligonucleotide, RNA, or DNA), a small molecule, derivatives thereof and analogs thereof.

[0072] The terms “peptide”, “polypeptide” and “protein” are used herein interchangeably to describe a series of at least two amino acids covalently linked by peptide bonds or modified peptide bonds such as isosteres. No limitation is placed on the maximum number of amino acids which may comprise a peptide or protein. Furthermore, the term polypeptide extends to fragments, analogues and derivatives of a peptide, wherein said fragment, analogue or derivative retains the same biological functional activity as the peptide from which the fragment, derivative or analogue is derived. A polypeptide as used herein may be encoded by a recombinant nucleic acid.

[0073] A “fragment” is a portion of a protein or nucleic acid that is substantially identical to a reference protein or nucleic acid. In some embodiments, the portion retains at least 50%, 75%, or 80%, or 90%, 95%, or even 99% of the biological activity of the reference protein or nucleic acid described herein.

[0074] A "functional derivative" or “functional fragment” of a native sequence polypeptide (e.g., antibody) is a compound having a qualitative biological property in common with a native sequence polypeptide. Functional derivatives or fragments include, but are not limited to, fragments of a native sequence and derivatives of a native sequence polypeptide and its fragments, provided that they have a biological activity in common with a corresponding native sequence polypeptide. The terms "derivative" and “fragment” encompass both amino acid sequence variants of polypeptide and covalent modifications thereof. Attorney Docket No.: S199207 1060WO (00093)

[0075] ST-020-W01

[0076] As used herein, the term “antibody” refers to any form of immunoglobulin molecule that exhibits the desired biological or binding activity. Thus, it is used in the broadest sense and specifically covers, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), humanized, fully human antibodies, and chimeric antibodies, and may include post-translational modifications thereof (e.g., C-terminal Lysine clipping in the heavy chain, conversion of glutamine or glutamic acid to pyroglutamate) that may occur when an antibody is recombinantly expressed in host cells (e.g., CHO cells), or during purification / storage. “Parental antibodies” are antibodies obtained by exposure of an immune system to an antigen prior to modification of the antibodies for an intended use, such as humanization of an antibody for use as a human therapeutic. As used herein, the term “antibody” encompasses not only intact polyclonal or monoclonal antibodies, but also, unless otherwise specified, fusion proteins comprising an antigen binding fragment thereof that competes with the intact antibody for specific.

[0077] In general, the basic antibody structural unit comprises a tetramer. Each tetramer includes two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The variable regions of each light / heavy chain pair form the antibody binding site. Thus, in general, an intact antibody has two binding sites. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Typically, human light chains are classified as kappa and lambda light chains. Furthermore, human heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and define the antibody’s isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, N.Y. (1989).

[0078] “Variable regions” or “V region” or “V chain” as used herein means the segment of IgG chains which is variable in sequence between different antibodies. A “variable region” of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. The variable region of the heavy chain may be referred to as “VH.” The variable region of the light chain may be referred to as “VL.”

[0079] Typically, the variable regions of both the heavy and light chains comprise three hypervariable regions, also called complementarity determining regions (CDRs), which are located within relatively conserved framework regions (FR). The CDRs are usually aligned by Attorney Docket No.: S199207 1060WO (00093)

[0080] ST-020-W01 the framework regions, enabling binding to a specific epitope. In general, from N-terminal to C- terminal, both light and heavy chains variable domains comprise FR1 , CDR1 , FR2, CDR2, FR3, CDR3, and FR4. As referred to herein the light chain CDRs are CDRL1 , CDRL2 and CDRL3, respectively, and the heavy chain CDRs are CDRH1 , CDRH2 and CDRH3, respectively. The assignment of amino acids to each domain is, generally, in accordance with the definitions of Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32:1-75; Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616; Chothia, et al., (1987) J Mol. Biol. 196:901- 917 or Chothia, et al., (1989) Nature 342:878-883.

[0081] A “CDR” refers to one of three hypervariable regions (H1 , H2, or H3) within the nonframework region of the antibody VH p-sheet framework, or one of three hypervariable regions (L1 , L2, or L3) within the non-framework region of the antibody VL p-sheet framework. Accordingly, CDRs are variable region sequences interspersed within the framework region sequences. CDR regions are well known to those skilled in the art and have been defined by, for example, Kabat as the regions of most hypervariability within the antibody variable domains. CDR region sequences also have been defined structurally by Chothia as those residues that are not part of the conserved p-sheet framework, and thus are able to adapt to different conformations. Both terminologies are well recognized in the art. CDR region sequences have also been defined by AbM, Contact, and IMGT. The positions of CDRs within a canonical antibody variable region have been determined by comparison of numerous structures (Al- Lazikani et al., 1997, J. Mol. Biol. 273:927-48; Morea et al., 2000, Methods 20:267-79). Because the number of residues within a hypervariable region varies in different antibodies, additional residues relative to the canonical positions are conventionally numbered with a, b, c and so forth next to the residue number in the canonical variable region numbering scheme (Al-Lazikani et al., supra). Such nomenclature is similarly well known to those skilled in the art. Correspondence between the numbering system, including, for example, the Kabat numbering and the IMGT unique numbering system, is well known to one skilled in the art and shown below in Table 1. In some embodiments, the CDRs are as defined by the Kabat numbering system. In other embodiments, the CDRs are as defined by the IMGT numbering system. In yet other embodiments, the CDRs are as defined by the AbM numbering system. In still other embodiments, the CDRs are as defined by the Chothia numbering system. In yet other embodiments, the CDRs are as defined by the Contact numbering system. Attorney Docket No.: S199207 1060WO (00093)

[0082] ST-020-W01

[0083] Correspondence between the CDR Numbering Systems

[0084] TABLE 1

[0085] “Chimeric antibody” refers to an antibody in which a portion of the heavy and / or light chain contains sequences derived from a particular species (e.g., human) or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is derived from another species (e.g., mouse) or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity.

[0086] “Human antibody” refers to an antibody that comprises human immunoglobulin protein sequences or derivatives thereof. A human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “rat antibody” refer to an antibody that comprises only mouse or rat immunoglobulin sequences or derivatives thereof, respectively.

[0087] “Humanized antibody” refers to forms of antibodies that contain sequences from nonhuman (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from non-human immunoglobulin. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The prefix “hum”, “hu” or “h” may be added to antibody clone designations when necessary to distinguish humanized Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 antibodies from parental rodent antibodies. The humanized forms of rodent antibodies will generally comprise the same CDR sequences of the parental rodent antibodies, although certain amino acid substitutions may be included to increase affinity, increase stability of the humanized antibody, or for other reasons.

[0088] “Monoclonal antibody” or “mAb” or “Mab”, as used herein, refers to a population of substantially homogeneous antibodies, i.e. , the antibody molecules comprising the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include a multitude of different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are often specific for different epitopes. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present disclosure may be made by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597, for example. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0089] As used herein, unless otherwise indicated, “antibody fragment” or “antigen binding fragment” refers to a fragment of an antibody that retains the ability to bind specifically to the antigen, e.g., fragments that retain one or more CDR regions and the ability to bind specifically to the antigen. An antibody that “specifically binds to” CD20 is an antibody that exhibits preferential binding to CD20 (as appropriate) as compared to other proteins, but this specificity does not require absolute binding specificity. An antibody is considered “specific” for its intended target if its binding is determinative of the presence of the target protein in a sample, e.g., without producing undesired results such as false positives. Antibodies, or binding fragments thereof, will bind to the target protein with an affinity that is at least two-fold greater, preferably at least ten times greater, more preferably at least 20-times greater, and most preferably at least 100-times greater than the affinity with non-target proteins.

[0090] Antigen binding portions include, for example, Fab, Fab’, F(ab’)2, Fd, Fv, fragments including CDRs, and single chain variable fragment antibodies (scFv), and polypeptides that contain at least a portion of an immunoglobulin that is sufficient to confer specific antigen binding to the antigen (e.g., CD20). An antibody includes an antibody of any class, such as IgG, Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0091] IgA, or IgM (or sub-class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant region of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., lgG1, lgG2, lgG3, lgG4, lgA1 , and lgA2. The heavy-chain constant regions that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0092] An "antigen" is a structure to which an antibody can selectively bind. A target antigen may be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. In some embodiments, the target antigen is a polypeptide. In certain embodiments, an antigen is associated with a cell, for example, is present on or in a cell, for example, a cancer cell or an immune cell.

[0093] An "intact" antibody is one comprising an antigen-binding site as well as a constant domain (CL) and at least heavy chain constant regions, CH1 , CH2 and CH3. The constant regions may include human constant regions or amino acid sequence variants thereof. In certain embodiments, an intact antibody has one or more effector functions.

[0094] As used herein, the term “immune response” relates to any one or more of the following: specific immune response, non-specific immune response, both specific and non-specific response, innate response, primary immune response, adaptive immunity, secondary immune response, memory immune response, immune cell activation, immune cell-proliferation, immune cell differentiation, and cytokine expression.

[0095] “Chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Classes of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, kinase inhibitors, spindle poison plant alkaloids, cytoxic / antitumor antibiotics, topoisomerase inhibitors, photosensitizers, anti-estrogens and selective estrogen receptor modulators (SERMs), anti-progesterones, estrogen receptor down-regulators (ERDs), estrogen receptor antagonists, leutinizing hormone-releasing hormone agonists, anti-androgens, aromatase inhibitors, EGFR inhibitors, VEGF inhibitors, and anti-sense oligonucleotides that inhibit expression of genes implicated in abnormal cell proliferation or tumor growth. Chemotherapeutic agents useful in the treatment methods of the invention include cytostatic and / or cytotoxic agents.

[0096] A "biological sample" encompasses a variety of sample types obtained from an individual or a population of individuals and can be used in a diagnostic, monitoring or screening Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 assay. The definition encompasses blood and other liquid samples of biological origin, solid tissue samples such as a biopsy specimen or tissue cultures or cells derived therefrom and the progeny thereof. The definition also includes samples that have been manipulated in any way after their procurement, such as by mixing or pooling of individual samples, treatment with reagents, solubilization, or enrichment for certain components, such as cells, polynucleotides, polypeptides, etc. The term "biological sample" encompasses a clinical sample, and also includes cells in culture, cell supernatants, cell lysates, serum, plasma, biological fluid, and tissue samples. The term "biological sample" includes urine, saliva, cerebrospinal fluid, interstitial fluid, ocular fluid, synovial fluid, blood fractions such as plasma and serum, and the like. The term "biological sample" also includes solid tissue samples, tissue culture samples, and cellular samples.

[0097] “Cytokine Release Syndrome” or CRS, as used herein, refers to an acute systemic inflammatory syndrome that can be triggered by a variety of factors such as infections and may occur after treatment with some types of immunotherapy, such as monoclonal antibodies and chimeric antigen receptor (CAR) T cell therapies, as well as some non-protein-based cancer drugs. CRS is characterized by a large, rapid increase in cytokines and inflammatory response. Signs and symptoms of CRS include fever, fatigue, nausea, headache, rash, arthralgia, myalgia, rapid heartbeat, hypotension, and trouble breathing. CRS can progress to an uncontrolled systemic inflammatory response with vasopressor-requiring circulatory shock, vascular leakage, disseminated intravascular coagulation, and multi-organ system failure. Patients may have a mild reaction, or reactions may be severe or life threatening. Grading of CRS can be performed according to the Consensus American Society for Transplantation and Cellular Therapy (ASTCT) grading system, such as e.g., that described in Lee et al. Biology of Blood and Marrow Transplantation. 25 (2019) 625 - 639; the disclosure of which is incorporated herein by reference in its entirety.

[0098] Detailed Description

[0099] Provided are methods, and reagents and kits for performing such methods, directed to treating subjects with cancer, including subjects with hematological cancers and subjects with solid tumors, including subjects with relapsed and / or refractory hematological cancers, subjects with metastatic, recurrent, relapsed, unresectable, and / or advanced tumors. Provided are methods of treating cancer comprising administering a decoy-resistant interleukin 18 (DR-18), including e.g., using combination therapies comprising administering a DR-18 and a multispecific therapeutic. Methods may also include selecting a patient, and administering a DR-18 Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 to result in enhanced cytotoxic activity of immune cells in the patient to treat the subject for cancer. Also provided are methods for administration of DR-18, to prime immune cells present in the subject, and administration of one or more agents that activate the primed immune cells, resulting in enhanced cytotoxic activity, thereby treating the cancer.

[0100] 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.

[0101] 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.

[0102] 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.

[0103] 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.

[0104] 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 Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0105] 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.

[0106] 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.

[0107] 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.

[0108] Methods

[0109] Methods described herein involve the use of decoy-resistant interleukin 18 (DR-18) polypeptides, and / or DR-18 polypeptide encoding nucleic acids, for the treatment of subjects having cancer. Methods of the present disclosure may involve the selection of particular subjects for treatment using DR-18 polypeptides and / or nucleic acids encoding DR-18 polypeptides. Methods of the present disclosure may involve the use of DR-18 polypeptides, and / or DR-18 encoding nucleic acids, for the treatment of subjects having a hematological cancer, such as e.g., a leukemia, a lymphoma, or a myeloma. Methods of the present disclosure may involve the use of DR-18 polypeptides, and / or DR-18 encoding nucleic acids, for the treatment of subjects having a solid tumor cancer.

[0110] Methods of the present disclosure may involve the use of DR- 18 polypeptides, and / or DR-18 encoding nucleic acids, for the treatment of relapsed, refractory, recurrent, unresectable, and / or advanced cancers, such as relapsed hematological cancer, recurrent solid tumors, refractory hematological cancer, refractory solid tumors, unresectable solid tumors, advanced Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 solid tumors, and the like. Methods of the present disclosure may involve combination therapies employing a DR-18 polypeptide and / or a nucleic acid encoding a DR-18 polypeptide in combination with a multi-specific therapeutic, such as a bispecific or trispecific antibody.

[0111] Useful multi-specific therapeutics include, e.g., multi-specific therapeutics, wherein the multi-specific therapeutic comprises at least a first antigen binding region that specifically binds to a first antigen and at least a second binding region that specifically binds to a second antigen. In some instances, the first or second antigen is an antigen expressed on cancer cells. In some instances, the first or second antigen is an antigen expressed on immune cells within the subject. Useful multi-specific therapeutics include, e.g., multi-specific therapeutics, wherein the multi-specific therapeutic comprises at least a first antigen binding region that specifically binds to an antigen expressed on cells of the cancer and at least a second binding region that specifically binds to an antigen expressed on immune cells within the subject. For brevity, references herein to use or administration or treatment with a DR- 18, DR- 18 polypeptide, or DR- 18 therapeutic will also generally refer to the use or administration or treatment with a DR-18 encoding nucleic acid (unless clearly inapplicable or specifically indicated otherwise). Similarly, references to co-administration of a DR-18 or a DR-18 therapeutic will generally also serve as description referring to co-administration of a DR-18 encoding nucleic acid (unless clearly inapplicable in the context or specifically indicated otherwise).

[0112] As summarized above, methods described herein may involve selecting and treating subjects with hematological malignancies. In some embodiments, the methods involve selecting subjects for receiving administration of DR-18 based on the subjects having a reduced ability to produce anti-drug antibodies (ADAs) and neutralizing antibodies (nAbs) against the administered DR-18. In some embodiments, methods of the present disclosure may include selecting a particular subject having cancer, such as a hematological malignancy, for treatment with DR-18, such as e.g., a subject with a hematological malignancy and a reduced ability to produce ADAs and nAbs against the DR-18.

[0113] Methods may further comprise administering the subject a combination therapy that includes administering to the subject a DR-18 and a multi-specific therapeutic, such as e.g., a bispecific or trispecific antibody, that comprises at least a first antigen binding region that specifically binds to an antigen expressed on cells of the cancer and at least a second binding region that specifically binds to an antigen expressed on immune cells within the subject. In some instances, an antigen expressed on a cancer cell is an antigen expressed on a cancer cell of a hematological cancer, such as e.g., an antigen expressed on a lymphoma, a leukemia, or a Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 myeloma. In some instances, an antigen expressed on a cancer cell is an antigen expressed on a cancer cell of a solid tumor.

[0114] IL- 18 is a pro-inflammatory cytokine that can stimulate T cells, NK cells, and myeloid cells. IL-18 has been proposed as an immunotherapeutic agent for the treatment of cancer, given its ability to stimulate anti-tumor cells. IL-18 binds, and signals through, IL-18 receptor (IL- 18R; also referred to as Interleukin-18 receptor 1 , IL18R1 , IL-18R-1 , IL-18R1 , IL1 receptor- related protein, IL1 RRP, Interleukin-18 receptor alpha, IL-18Ra, and CD218a). Binding of IL-18 to IL-18Ra allows for formation of the IL-18 / IL-18Ra / IL-18R|3 complex which results in activation of IL-18 signaling and simulation of immune cells. IL-18 binding to IL-18R, and thus signaling through the receptor, may be blocked by IL-18 binding protein (IL-18BP), which has limited the clinical efficacy of recombinant human wild-type (WT) IL-18.

[0115] As described in more detail below, decoy-resistant IL-18 (DR-18) polypeptides bind to, and signal through formation of, the IL-18 receptor complex. DR-18 polypeptides do not bind to IL-18 binding protein (IL-18BP), or at least display substantially reduced binding to IL-18BP, such as substantially reduced binding to IL-18BP as compared to wild-type (WT) human IL-18 (SEQ ID NO: 1) binding IL-18BP.

[0116] Substantially reduced binding to IL-18BP may be characterized as a dissociation constant (KD) between DR-18 and IL-18BP of at least 10 nM or greater, such as e.g., 20 nM or greater, 30 nM or greater, 40 nM or greater, 50 nM or greater, 60 nM or greater, 70 nM or greater, 80 nM or greater, 90 nM or greater, 100 nM or greater, 200 nM or greater, 300 nM or greater, 400 nM or greater, 500 nM or greater, 600 nM or greater, 700 nM or greater, 800 nM or greater, 900 nM or greater, or 1 pM or greater. Affinity in general, and KD specifically, may be measured by any convenient and appropriate method including but not limited to e.g., spectroscopic assays, titration calorimetry, or using optical biosensors, such as in Surface Plasmon Resonance (SPR). In some instances, binding between a DR-18 and IL-18BP is undetectable. Further description of DR-18 polypeptides with substantially reduced binding to IL-18BP can be found in W02019 / 051015, the disclosure of which is incorporated herein by reference in its entirety.

[0117] Human WT IL-18 is expressed initially in a pre-processed form (SEQ ID NO: 65) that includes a 36 amino acid propeptide (SEQ ID NO: 66). Unless clearly intended otherwise, references to WT IL- 18 polypeptides and DR-18 polypeptides will generally refer to the active mature forms of the polypeptides (i.e. , without an IL-18 propeptide). For example, reference to a DR-18 polypeptide having X mutations relative to WT human IL-18 shall, unless specifically Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 indicated otherwise, refer to X mutations occurring within amino acids 1 to 157 of SEQ ID NO: 1 (i.e. , amino acids 37 to 193 of SEQ ID NO: 65).

[0118] During clinical trials using an exemplary human DR-18, (referred to herein in the Examples as "Test Compound"), it was unexpectedly observed that a large subset of subjects demonstrated a marked and consistent production of anti-drug antibodies (ADAs) and, eventually, neutralizing antibodies (nAbs) (see Examples). This observation was particularly surprising given the high degree of amino acid sequence similarity (greater than 90% sequence identity) between the DR-18 and the active, processed form of wildtype human IL-18 (SEQ ID NO: 1).

[0119] The presence of these antibodies affected the bioavailability of administered DR-18 Test Compound, accelerating its clearance from the circulation and reducing the effective plasma concentration of the drug, following administration by subcutaneous injection. An impact on bioavailability of the administered DR-18 Test Compound by the presence of ADAs and / or nAbs may or may not influence the efficacy of the compound in the clinical setting in which it is tested. Nevertheless, this immunogenic response to the DR-18 Test Compound in patients was unanticipated and presented a complexity in the therapeutic application of the Test Compound and other DR-18 agents.

[0120] Without being bound by theory, it was surmised that use of a DR- 18, such as the Test Compound, in subjects with a repressed ability to mount an immune response to the drug and a having reduced ability to generate anti-DR-18 ADAs and nAbs would result in improved therapeutic outcomes. Subsequent analyses focused on identifying subjects who exhibit different levels of ADA and nAb production. Subjects with a compromised immune system, often a characteristic of hematological malignancies in a relapsed or refractory state and / or a characteristics of subjects having undergone treatment for a hematological malignancy, have a diminished ability to generate ADAs and / or nAbs. This reduced immunogenic response, attributed to impaired immune competence, provides certain advantages for application of DR- 18 treatments.

[0121] Building on the insights from the Test Compound trials, the administration of DR-18 has been improved by selecting subjects who are deficient in the production of ADAs and / or nAbs. This patient selection enhances, including substantially enhances, the method, including e.g., by increasing therapeutic efficacy of DR-18 (of a particular administered dose or series of doses) by mitigating the risk of immune-mediated neutralization and clearance of the therapeutic agent. In some instances, such selection enhances, including substantially enhances, the method, including e.g., by reducing the dose of DR-18 needed to elicit the desired response. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0122] In addition, further studies as described herein have shown that DR-18 administration can improve the effectiveness and / or durability of administered immunotherapeutics in cancer, including in hematological malignancies and solid tumors. For example, in some instances, coadministration of DR-18 and an immunotherapeutic, such as a multi-specific (e.g., bispecific) antibody, reduces growth of a cancer, improves duration of response to treatment in a cancer, or a combination thereof. In some instances, co-administration of DR-18 and an immunotherapeutic, such as a multi-specific (e.g., bispecific) antibody, reduces growth of a hematological malignancy, improves duration of response to treatment in a hematological malignancy, or a combination thereof. In some instances, co-administration of DR-18 and an immunotherapeutic, such as a multi-specific (e.g., bispecific) antibody, reduces growth of a solid tumor, improves duration of response to treatment in a solid tumor, or a combination thereof.

[0123] In some instances, co-administration of DR-18 and an immunotherapeutic, such as a multi-specific (e.g., bispecific) antibody, reduces growth of a relapsed and / or refractory cancer, improves duration of response to treatment in a relapsed and / or refractory cancer, or a combination thereof. In some instances, co-administration of DR-18 and an immunotherapeutic, such as a multi-specific (e.g., bispecific) antibody, reduces growth of a relapsed and / or refractory hematological malignancy, improves duration of response to treatment in a relapsed and / or refractory hematological malignancy, or a combination thereof. In some instances, co- administration of DR-18 and an immunotherapeutic, such as a multi-specific (e.g., bispecific) antibody, reduces growth of a relapsed and / or refractory solid tumor, improves duration of response to treatment in a relapsed and / or refractory solid tumor, or a combination thereof.

[0124] In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of relapse, improves response, improves the duration of response, and / or reduces at least one symptom of a leukemia. In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of relapse, improves response, improves the duration of response, and / or reduces at least one symptom of an AML. Factors affecting a patient’s risk of relapse for AML include age, AML subtype, cytogenetic and molecular abnormalities, level of remission achieved, MRD status. Patients are generally categorized into a risk group (low, intermediate, or high / adverse / poor) to guide further treatment decisions. In some instances, co-administration of DR-18 and an immunotherapeutic reduces risk of relapse by positively influencing, e.g., the level of remission achieved and / or MRD status, such that the patient is categorized into a lower risk category (e.g., as compared to the risk category that would be applied absent the co-administration). Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0125] In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of relapse, improves response, improves the duration of response, and / or reduces at least one symptom of a lymphoma. In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of relapse, improves response, improves the duration of response, and / or reduces at least one symptom of a BCL, including e.g., LBCL or DLBCL. Factors affecting a patient’s risk of relapse for BCL include age, disease / molecular subtype, lactate dehydrogenase (LDH) levels, stage of the disease, involvement of multiple extra-nodal sites, and time to relapse / refractory disease. Various tools are used by health care providers to assess risk of relapse in BLC patients, including e.g., international prognostic index (I PI) scoring, imaging biomarkers, genetic markers, liquid biopsies, and the like. In some instances, co-administration of DR-18 and an immunotherapeutic reduces risk of relapse by positively influencing, e.g., the stage of the disease and / or the time to relapse / refractory disease, such that the patient is categorized into a lower risk category (e.g., by I PI or other measures) (as compared to the risk category that would be applied absent the co-administration).

[0126] In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of relapse, improves response, improves the duration of response, and / or reduces at least one symptom of a myeloma. In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of relapse, improves response, improves the duration of response, and / or reduces at least one symptom of a multiple myeloma. Factors affecting a patient’s risk of relapse for myeloma include cytogenetic abnormalities, gene expression, clinical presentation, genetic mutations, biochemical markers, and response to treatment. Patients are generally categorized into a standard-risk group or a high-risk group. In some instances, co-administration of DR-18 and an immunotherapeutic reduces risk of relapse by positively influencing, e.g., gene expression, clinical presentation / response to treatment, and / or biochemical markers, such that the patient is categorized into a lower risk category (e.g., as compared to the risk category that would be applied absent the co-administration).

[0127] In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of recurrence, improves response, improves the duration of response, and / or reduces at least one symptom of a solid tumor. In some instances, co-administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of recurrence, improves response, improves the duration of response, and / or reduces at least one symptom of a melanoma or a colon cancer, such as a colon adenocarcinoma. In some instances, co- Attorney Docket No.: S199207 1060WO (00093)

[0128] ST-020-W01 administration of DR-18 and an immunotherapeutic reduces growth, reduces the risk of recurrence, improves response, improves the duration of response, and / or reduces at least one symptom of a metastatic (e.g., a locally metastatic, a regionally metastatic, or a distantly metastatic) tumor, a recurrent solid tumor, a relapsed solid tumor, unresectable, and / or an advanced solid tumor, including e.g., a metastatic, recurrent, relapsed, unresectable, and / or advanced melanoma or a metastatic, recurrent, relapsed, unresectable, and / or advanced colorectal tumor, such as e.g., a metastatic, recurrent, relapsed, unresectable, and / or advanced colon adenocarcinoma.

[0129] Without being bound by theory, administration of DR-18, by virtue of signaling through the IL-18 receptor present on immune cells in the subject, primes the immune cells for further activation. Immune cell priming generally refers to the process by which dormant or inactive immune cells are prepared to respond more effectively to antigens. In some instances, such priming may involve the initial interaction of the immune cell with antigen and / or one or more endogenous molecular signals. In some instances, such priming may involve or be enhanced by the presence of DR-18 and its signaling through the IL-18 receptor. Primed immune cells display various characteristics, such as e.g., activation, proliferation, effector functions, costimulatory molecule expression, adhesion molecule expression, upregulation of signaling pathways that are downregulated in unprimed or exhausted cells, downregulation of signaling pathways that are upregulated in unprimed or exhausted cells, and the like. In some instances, primed cells may expression one or more activation markers, such as e.g., CD69 and / or CD25. In some instances, primed cells may display proliferative characteristics, such as e.g., expression of Ki67 or PCNA, labeling with proliferation labels / dyes, or the like. In some instances, primed cells may display effector functions, such as e.g., the secretion of one or more cytokines. In some instances, primed cells may express or up-regulate one or more costimulatory markers, such as e.g., CD40L. In some instances, primed cells may express of up- regulate one or more adhesion molecules, such as e.g., LFA-1. These characteristics of primed immune cells function, at least in part, to aid in interaction with other immune cells and the full activation of the immune cell upon exposure to additional antigen. Such characteristics of primed immune cells may be detected by any convenient and appropriate means, including e.g., flow cytometry, PCR, NGS, gene expression analyses, proliferation assays, cytokine assays, and the like.

[0130] In some instances, a primed immune cell may be further primed or further activated by interaction with DR-18. In some instances, an immune cell that has been primed by interaction with DR- 18 may be further activated by interaction with a co-administered therapeutic. For Attorney Docket No.: S199207 1060WO (00093)

[0131] ST-020-W01 example, bispecific T cell engagers function by increasing engagement between an immune cell and a cancer. In some instances, an immune cells is contacted with a DR-18 thereby priming the immune cell and the primed immune cell is engaged with a cancer cell through interaction with the bispecific T cell engager, resulting in cytotoxic activity towards the cancer cell. Interaction of a primed to cell with a cancer cell (e.g., through the use of a bispecific T cell engager) results in enhanced cytotoxicity towards the cancer cell as compared to the level of cytotoxicity observed in the absence of priming with DR-18. Such enhanced cytotoxicity activity may manifest in a higher level of cell killing, a longer duration of cell killing, or a combination thereof.

[0132] Methods of the present disclosure need not necessarily employ a co-administered immunotherapeutic to further activate a primed immune cell. In some instances, a primed immune cell may be further activated, e.g., through interaction with antigen present on cancer cells within the subject, through interaction with other immune cells present within the subject, interaction with signaling molecules (e.g., cytokines) present within the subject, or any combination thereof. In some instances, employing a DR-18 according to the methods as described herein enhances immune cell priming and / or activation, e.g., as compared to priming and / or activation by cancer cells, immune cells, and / or endogenous cytokines in the absence of DR-18.

[0133] Methods described herein may involve selecting subjects for DR-18 administration based on their impaired ability to produce ADAs and / or nAbs. In some embodiments, subjects with relapsed and / or refractory hematological cancers, who typically exhibit compromised immune functions, are identified as suitable candidates. These subjects demonstrate a reduced propensity to mount an immunogenic response against DR-18, thereby allowing for sustained therapeutic activity and improved clinical outcomes. In some embodiments, a subject with an impaired ability to produce ADAs and / or nAbs is a subject that has been previously treated with one or more cancer therapies (e.g., chemotherapy, radiation therapy, immunotherapy, or any combination thereof) resulting in compromised immune functions in the subject. In some embodiments, the subject has been previously treated with the standard of care (SOC) therapy for their cancer prior to administering a DR-18 as described herein.

[0134] In some embodiments, a subject treated in the herein described methods is selected based on having a relapsed cancer, including a relapsed hematological malignancy or a relapsed solid tumor. In some embodiments, a subject treated in the herein described methods is selected based on having a refractory cancer, including a refractory hematological malignancy or a refractory solid tumor. In some embodiments, a subject treated in the herein Attorney Docket No.: S199207 1060WO (00093)

[0135] ST-020-W01 described methods is selected based on having a reduced number and / or activity of immune cells, including e.g., reduced number and / or activity of B cells, reduced number or activity of plasma cells, etc. In some instances, the reduced ability of the subject to generate ADA and / or nAb is a result of the subject's cancer and / or prior cancer treatment. In some instances, the decreased number and / or activity of immune cells, such as B cells and / or plasma cells, of the subject is a result of the subject's cancer and / or prior cancer treatment.

[0136] In some instances, the reduced ability of the subject to generate ADA and / or nAb is not a result of the subject having been administered obinutuzumab or some other B cell depleter. In some instances, the decreased number and / or activity of immune cells, such as B cells and / or plasma cells, of the subject is not a result of the subject having been administered obinutuzumab or some other B cell depleter. In some instances, the subject is not administered (as part of the method), or has not been administered (prior to and / or during the method), obinutuzumab or another B cell depleter.

[0137] In some instances, prior to administration of a DR- 18, a subject treated according to a herein described method may have received a prior leukapheresis and / or other lymphodepleting treatment. Leukapheresis is an apheresis procedure that involves the separation and removal of white blood cells from a patient's blood and the re-infusion of leukocyte-poor plasma. See e.g., Zhang et al. IJGM (2021) 14: 3457-3467. Other useful lymphodepleting treatments include e.g., lymphodepleting chemotherapy. In some instances, lymphodepleting chemotherapy is employed prior to adoptive cell therapies, CAR T cell therapies, and the like. Protocols for lymphodepleting chemotherapy vary, but common current standard protocols generally involve a combination of cyclophosphamide and fludarabine, sometimes followed by radiation therapy, such as e.g., total body irradiation (TBI).

[0138] In some instances, a subject treated according to a method described herein may have undergone a leukapheresis and / or lymphodepleting chemotherapy pretreatment prior to administration of DR-18. In some instances, a subject receives, or has received, leukapheresis at least 3 days prior, at least 5 days prior, at least 7 days prior, at least 10 days prior, at least 12 days prior, at least 14 days prior to administration of DR-18. In some instances, a subject receives, or has received, lymphodepleting chemotherapy at least 3 days prior, at least 5 days prior, at least 7 days prior, at least 10 days prior, at least 12 days prior, at least 14 days prior to administration of DR-18.

[0139] Certain benefits of selective patient population selection in DR-18 therapy include, but are not limited to, enhanced therapeutic efficacy, reduced adverse effects, and improved dosing regimens. For example, without being bound by theory, enhanced therapeutic efficacy may be Attorney Docket No.: S199207 1060WO (00093)

[0140] ST-020-W01 achieved by reducing the likelihood of immune-mediated neutralization, such that DR-18 maintains its therapeutic activity over the treatment course, resulting in enhanced cytotoxic activity of anti-cancer immune cells. As used herein, cytotoxic activity refers to the process by which immune cells, such as cytotoxic T cells (e.g., CD8+ T cells) and / or Natural Killer (NK) cells directly kill cancerous cells. Accordingly, cytotoxic activity may be enhanced in that cancer cell killing is increased due to the presence of DR-18, e.g., as compared to the level of cancer cell killing observed or expected in a corresponding setting in the absence of DR-18.

[0141] Markers of cytotoxic activation of T cells include e.g., CD25, CD69, CD71 , HLA-DR, CD154, Perforin, Granzyme B, and Interferon-gamma (IFN-y). In some instances, enhanced cytotoxic activity may be detected through the detection or measurement of one or more markers of cytotoxic activation on or in T cells, including e.g., where the marker (or more than one of the markers) is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 800%, or 1000% or more as compared to the expression of the marker in on or in corresponding T cells not treated with DR- 18 and / or corresponding resting T cells. Levels of such markers may, where appropriate, be assessed by measuring the protein and / or gene expression level of the marker where any convenient and appropriate assay may be employed, including e.g., enzyme linked immunosorbent assay (ELISA), flow cytometry, PCR, next generation sequencing (NGS), mass spectrometry, and the like.

[0142] Markers of cytotoxic activation of NK cells include e.g., CD69, CD25, Granzyme B, Perforin, IFN-y, Killer Immunoglobulin-like Receptors (KIRs), Natural Cytotoxicity Receptors (NCRs), NKG2D (KLRK1), CD16, NKp46, NKp30, and NKp44. In some instances, enhanced cytotoxic activity may be detected through the detection or measurement of one or more markers of cytotoxic activation on or in NK cells, including e.g., where the marker (or more than one of the markers) is increased at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 800%, or 1000% or more as compared to the expression of the marker in on or in corresponding NK cells not treated with DR-18 and / or corresponding resting NK cells. Levels of such markers may, where appropriate, be assessed by measuring the protein and / or gene expression level of the marker where any convenient and appropriate assay may be employed, including e.g., ELISA, flow cytometry, PCR, NGS, mass spectrometry, and the like.

[0143] In some instances, enhanced cytotoxic activity of immune cells may be detected and / or measured through the presence or secretion of one or more immune activating cytokines. Nonlimiting examples of immune activating cytokines include e.g., Interferon-gamma (IFN-y), Tumor Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0144] Necrosis Factor-alpha (TNF-a), lnterleukin-2 (IL-2), Interleukin-12 (IL-12), Granulocyte- Macrophage Colony-Stimulating Factor (GM-CSF), Interleukin-15 (IL-15), and the like. Cytokines may be detected, and / or the level measured, in a sample by any convenient means, including but not limited to, e.g., ELISA, mass spectroscopy, and the like. In some instances, enhanced cytotoxic activity may be detected through the detection or measurement of an increase in one or more cytokines, including e.g., where the cytokine(s) is(are) increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 800%, or 1000% or more as compared to the level of cytokine in a sample from corresponding immune cells expression not treated with DR-18 and / or corresponding resting immune cells.

[0145] Cytotoxic activity may also be measured in vitro or ex vivo via a cell killing assay where the potentially cytotoxic cells are co-cultured with target cells, e.g., cancer cells, and survival of the cancer cells is assessed. In some instances, enhanced cytotoxic activity may be measured by an in vitro or in vivo cell killing assay where cell killing may be increased by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 800%, or 1000% or more as compared to the level of cell killing by corresponding cells not treated with DR-18. In some instances, enhanced cytotoxic activity may be inferred by effects observed through in vivo activity of the agent(s), including e.g., through observed inhibition of tumor growth, inhibition of the proliferation of blood cancer cells, reduction in tumor size, reduction in the number of blood cancer cells, and the like.

[0146] In some instances, cytotoxic activity may be enhanced, through the methods described herein, by increasing the duration of activity of cytotoxic immune cells, i.e. , increasing the length of time during which the cells maintain cytotoxic activity. Put another way, the duration of cytotoxic activity may be increased by preventing or delaying immune cell exhaustion. Cytotoxic activity may be enhanced, through the methods described herein, by increasing the amount of time the immune cells display cytotoxic activity (e.g., as measured by one or more of the foregoing techniques) by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% or more as compared to the duration of cytotoxic activity in corresponding immune cells not treated with DR-18.

[0147] Reduced Adverse Effects may, without being bound by theory, be achieved through the lowering of immunogenic response, which minimizes the risk of immune-related adverse effects, thereby improving the overall safety profile of the therapy. Improved dosing regimens may, without being bound by theory, be achieved because the administration of DR-18 can be tailored to the patient’s disease or immunological profile (with less to no impact of ADAs and / or Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 nAbs on bioavailability), potentially reducing the required dosage and / or frequency of administration.

[0148] The surprising discovery of ADA and nAb production in clinical trials, which represented the first ever administration of DR-18 to human subjects, underscores the advantages patient selection based on immunogenic profiles can provide. The selection of patients with a reduced ability to produce ADAs and / or nAbs enhances, among other elements of the therapy, the therapeutic efficacy of DR-18 in treating cancer, including hematological malignancies and solid tumors. The selection of patients with a reduced ability to produce ADAs and / or nAbs also enhances, among other elements of the therapy, the therapeutic efficacy of DR-18 in treating relapsed and refractory cancers, including hematological malignancies and solid tumors.

[0149] In some embodiments, subjects with hematological cancers exhibit a reduced ability to produce ADAs and nAbs to DR-18. This reduced immunogenic response can be attributed to the compromised immune function in these subjects. In some instances, those with refractory and / or relapsing hematological cancers have diminished immune capabilities, leading to a decreased production of ADAs and nAbs against the administered DR-18. This immunological deficiency improves the therapeutic efficacy of DR-18 by, at least in part, reducing the likelihood of an immune-mediated neutralization of the therapeutic agent.

[0150] In some embodiments, a subject with relapsed and / or refractory lymphoma is selected based on their immunological profile. Specifically, the subject's ability to produce ADAs and / or nAbs against the administered DR-18 is assessed. The subject demonstrates a diminished immunogenic response, characterized by lower ADA and / or nAb titers, resulting in enhanced therapeutic efficacy of DR-18.

[0151] In some embodiments, a subject with relapsed and / or refractory leukemia is selected based on their impaired immune function. The selection involves evaluating the subject's production of ADAs and / or nAbs to DR-18. The subject, due to their compromised immune system, exhibits a reduced ability to mount an immunogenic response, thereby allowing for improved bioavailability and sustained therapeutic activity of DR-18.

[0152] In some embodiments, a subject with relapsed and / or refractory myeloma is selected by determining their ADA and / or nAb levels against DR-18. The subject's reduced capacity to generate an immune response to the administered DR-18, resulting from the refractory nature of their hematological cancer and / or the prior treatment of their hematological cancer, leads to a lower risk of immune-mediated neutralization of the therapeutic agent. This selection enhances the clinical outcomes of DR-18 therapy in the treatment of myeloma. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0153] In some embodiments, a subject with a relapsed, recurrent, and / or refractory solid tumor is selected by determining their ADA and / or nAb levels against DR-18. The subject's reduced capacity to generate an immune response to the administered DR-18, resulting from prior treatment of their relapsed, recurrent, and / or refractory solid tumor, leads to a lower risk of immune-mediated neutralization of the therapeutic agent. This selection enhances the clinical outcomes of DR-18 therapy in the treatment of the relapsed, recurrent, and / or refractory solid tumor. In some instances, the solid tumor may be a colorectal tumor or a melanoma. In some instances, the solid tumor is a malignant, unresectable, and / or advanced solid tumor. In some instances, the solid tumor is a malignant, unresectable, and / or advanced colorectal tumor or a melanoma.

[0154] This subject identification or patient selection may include assessing the immunological profile of the subject to determine the level of ADAs and / or nAbs production. Subjects exhibiting diminished immunogenic response to DR-18, characterized by lower ADA and / or nAb titers, are deemed suitable candidates for DR-18 administration, thereby improving the therapeutic efficacy of the DR-18 treatment regimen. In some embodiments, an immunological profile may not be performed, e.g., the method may not include performing an immunological profile for the subject. In some embodiments, an ADA titer may not be performed, e.g., the method may not include performing an ADA titer for the subject. In some embodiments, a nAb titer may not be performed, e.g., the method may not include performing a nAb titer for the subject. In some instances, one or more assessments of the subject, as described herein, may have been previously performed to identify the subject (e.g., prior to performing a method involving administering DR-18 to the subject).

[0155] Methods of the present disclosure may include identifying, or having identified, a subject having a cancer that is relapsed and / or refractory to at least one SOC therapy for the cancer. For example, methods of the present disclosure may include identifying, or having identified, a subject having a hematological cancer that is relapsed and / or refractory to at least one SOC therapy for the hematological cancer. This identification may also include assessing whether the subject is deficient in the production of ADAs and / or nAbs, including where such assessment is performed following the SOC therapy. In some instances, such an identification may not include assessing whether the subject is deficient in the production of ADAs and / or nAbs. In some instances, subjects selected for treatment according to the methods described herein may be selected as having one or more of refractoriness, intolerance, or ineligibility to other established therapies known to provide clinical benefit for their cancer. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0156] In some embodiments, the current SOC for multiple myeloma involves initial induction therapy followed by consolidation and maintenance therapy. Historically, induction therapy often includes a combination of proteasome inhibitors (such as bortezomib), immunomodulatory drugs (such as lenalidomide), and steroids (such as dexamethasone) (Rajkumar (2020) Am J Hematol 95(5):548-567). In some embodiments, a subject treated according to a method described herein may have been previously treated with induction therapy.

[0157] In some embodiments, subjects treated according to a method as described herein may include subjects having multiple myeloma and having had at least one, at least two, at least three, or at least four prior lines of therapy that included one or more of a proteasome inhibitor, immune modulatory imide drug, and CD38 monoclonal antibody. In some embodiments, such subject will also have one or more of refractoriness, intolerance, or ineligibility to other established therapies known to provide clinical benefit for multiple myeloma.

[0158] In some embodiments, high-dose chemotherapy with autologous stem cell transplantation (ASCT) is utilized for eligible patients after achieving an adequate response to induction therapy. Melphalan is a common agent used in high-dose chemotherapy in this context (Srour et al. (2021) Haematologica 106(12):3211—3214). In some embodiments, a subject treated according to a method described herein may have been previously treated with high-dose chemotherapy plus ASCT. For example, in some instances a subject may have relapsed and / or refractory disease following ASCT. In some embodiments, methods of the present disclosure used to treat the subject do not comprise ASCT. In some instances, the therapy administered to the subject does not include ASCT, and / or subjects treated in the methods of the present disclosure are not administered ASCT during treatment.

[0159] In some embodiments, maintenance therapy, typically post-ASCT, includes continuing treatment with immunomodulatory drugs such as lenalidomide to prolong remission and improve overall survival rates (McCarthy et al. (2017) J Clin Oncol 35(29): 3279-3289). In some embodiments, a subject treated according to a method described herein may have been previously treated with maintenance therapy. In some instances, a subject may have relapsed and / or refractory disease following maintenance therapy.

[0160] In some embodiments, for patients who are not candidates for ASCT, combination regimens utilizing proteasome inhibitors, immunomodulatory drugs, and steroids remain the standard, with the addition of agents such as monoclonal antibodies (e.g., daratumumab) based on individual patient profiles and treatment responses (Palumbo et al. (2016) N Engl J Med 375(8): 754-66). In some embodiments, a subject treated according to a method described herein may have been previously treated with combination regimens utilizing proteasome Attorney Docket No.: S199207 1060WO (00093)

[0161] ST-020-W01 inhibitors, immunomodulatory drugs, and steroids, including with or without monoclonal antibodies, such as e.g., daratumumab. In some instances, a subject may have relapsed and / or refractory disease following treatment with one or more of proteasome inhibitors, immunomodulatory drugs, steroids, and monoclonal antibodies, such as e.g., daratumumab.

[0162] In some embodiments, SOC therapy for Acute Myeloid Leukemia (AML) may include the use of intensive chemotherapy regimens. Such regimens commonly involve a combination of cytarabine and an anthracycline, such as daunorubicin or idarubicin (Dohner et al., (2017) Blood 129(4):424-447). In some embodiments, a subject treated according to a method described herein may have been previously treated with an intensive chemotherapy regimen, including e.g., where the intensive chemotherapy regimen included cytarabine, an anthracycline (e.g., daunorubicin or idarubicin), or any combination thereof. In some instances, a subject may have relapsed and / or refractory disease following treatment with intensive chemotherapy. In some embodiments, prior treatment for AML may be intensive or non-intensive, including e.g., where such intensive and non-intensive therapies include those described in Jaramillo & Schlenk (2023) Haematologica. 108(2):342-352.

[0163] In some instances, a subject treated according to a method described herein is a subject with a leukemia or myelodysplastic syndrome that has returned or sustained prior treatment, including e.g., recurrent leukemia or myelodysplastic syndrome, persistent leukemia or myelodysplastic syndrome, or refractory leukemia or myelodysplastic syndrome. In some instances, a subject treated according to a method described herein is a subject with AML that has returned or sustained prior treatment, including e.g., recurrent AML, persistent AML, or refractory AML.

[0164] In some embodiments, the current SOC for a lymphoma, such as e.g., large B-cell lymphoma (LBCL) or diffuse large B-cell lymphoma (DLBCL) may generally involve generally involves chemotherapy and rituximab, with or without radiation therapy. For example, a SOC therapy for newly diagnosed or untreated DLBCL is the combination known as R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) given in 21-day cycles for an average of 6 cycles, however the length and number of cycles can vary. In some instances, a SOC therapy for DLBCL further includes etoposide. In some instances, a SOC therapy for DLBCL may include polatuzumab vedotin-piiq in combination with rituximab, cyclophosphamide, doxorubicin, and prednisone, with or without radiation therapy.

[0165] In some embodiments, subjects treated according to a method as described herein may include subjects having lymphoma and having had at least one, at least two, at least three, or at least four prior lines of therapy that included one or more of a SOC therapy described above Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 and / or one or more of platin-based induction followed by high-dose therapy (HDT; e.g., rituximab, carmustine, etoposide, cytarabine, and melphalan (R-BEAM) or rituximab, cyclophosphamide, etoposide, and carmustine (R-CVB)); platinum-based salvage chemoimmunotherapy with rituximab, gemcitabine, dexamethasone, and cisplatin (R-GDP); rituximab, ifosfamid, carboplatin, and etoposide (R-ICE); rituximab, dexamethasone, high-dose cytarabine, and cisplatin (R-DHAP); autologous stem cell transplantation (ASCT); anti-CD19 chimeric antigen receptor (CAR) T cell therapy (e.g., using axicabtagene ciloleucel, lisocabtagene maraleucel, or tisagenlecleucel); chemoimmunotherapy; monoclonal antibodies (e.g., tafasitamab + lenalidomide); antibody drug conjugates (e.g., Polatuzumab-vedotin + BR or Loncastuximab-tesirine); bispecific antibodies and BiTEs (e.g., blinatumomab, mosunetuzumab, glofitamab, or epcoritamab); immune checkpoint inhibitors (e.g., nivolumab or magrolimab); ibrutinib; lenalidomide; copanlisib; venetoclax; tazemetostat; selinexor; or the like. Useful SOC and other therapies include e.g., those described in Frontzek et al. (2022) Ther Adv Hematol. 13:20406207221103321. In some embodiments, besides SOC therapies such subjects will also have one or more of refractoriness, intolerance, or ineligibility to other established therapies known or thought to provide clinical benefit for lymphoma, such as LBCL or DLBCL.

[0166] In some instances, a subject treated according to a method described herein is a subject with a lymphoma that has returned or sustained prior treatment, including e.g., recurrent lymphoma, persistent lymphoma, or refractory lymphoma. In some instances, a subject treated according to a method described herein is a subject with LBCL or DLBCL that has returned or sustained prior treatment, including e.g., recurrent LBCL, persistent LBCL, refractory LBCL, recurrent DLBCL, persistent DLBCL, refractory DLBCL.

[0167] In some embodiments, post-remission therapy may be employed following initial induction therapy to prevent relapse. This therapy can include high-dose chemotherapy, consolidation therapy, or hematopoietic stem cell transplantation, depending on the patient's risk stratification and overall health status (Estey and Ddhner (2006) Lancet 368(9550): 1894- 907). In some embodiments, a subject treated according to a method described herein may have been previously treated with post-remission therapy. In some instances, a subject may have relapsed and / or refractory disease following post-remission therapy.

[0168] In some embodiments, targeted therapies may be integrated into the treatment plan, particularly for patients with specific genetic mutations. For example, FLT3 inhibitors, such as midostaurin, can be used in conjunction with standard chemotherapy for patients with FLT3- mutated AML (Stone et al. (2017) N Engl J Med 377(5) :454-464). In some embodiments, a subject treated according to a method described herein may have been previously treated with Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 one or more targeted therapies, including e.g., a FLT3 inhibitor, such as midostaurin. In some instances, a subject may have relapsed and / or refractory disease following one or more targeted therapies, including e.g., a FLT3 inhibitor, such as midostaurin.

[0169] In some embodiments, supportive care, including administration of anti-emetics, antibiotics, and transfusion support, may be critical to manage side effects and complications associated with intensive chemotherapy and to maintain patient well-being during treatment. In some embodiments, a subject treated according to a method described herein may have been previously treated with supportive care, including e.g., one or more of anti-emetics, antibiotics, and transfusion support, or any combination thereof.

[0170] In some embodiments, patient-specific factors such as age, comorbidities, and genetic profile may guide the customization of therapy, allowing for a personalized approach to treatment. In some embodiments, a subject treated according to a method described herein may have been previously treated with patient-specific customized therapy, such as e.g., a therapy based on the subject’s genetic profile.

[0171] Methods of the present disclosure also include administering a plurality of doses of a decoy-resistant interleukin 18 (DR-18) to the identified subject. The DR-18 is provided in an effective amount, such as e.g., an amount effective to activate an inactive anti-cancer immune cell population against the cancer, e.g., hematological cancer, thereby treating the subject for the cancer, e.g., for the hematological cancer. By "inactive", as used herein, is meant at least insufficiently active to treat the cancer. In some embodiments, DR-18 is provided in an effective amount, such as e.g., an amount effective to inhibit tumor growth, an amount effective to at least induce stable disease, an amount effective to at least induce a partial response, an amount effective to induce a complete response. In some embodiments, DR-18 is provided in an effective amount, such as e.g., an amount effective to prevent or at least slow the rate of growth of a tumor, such as a refractory tumor. Such amounts may be, e.g., an amount effective to achieve the desired result employing DR- 18 as a monotherapy or an amount effective to achieve the desired result when DR-18 is employed as part of a combination therapy, e.g., in combination with at least one multi-specific therapeutic.

[0172] The amount of DR-18 administered to a subject can vary, including e.g., where the amount of agent administered to a subject in a single dose can range from about 1 pg / kg (of body weight of the subject) or less to about 3000 pg / kg or more, including e.g., from about 1 pg / kg to about 3000 pg / kg, about 5 pg / kg to about 3000 pg / kg, about 10 pg / kg to about 3000 pg / kg, about 15 pg / kg to about 3000 pg / kg, from about 20 pg / kg to about 3000 pg / kg, from about 30 pg / kg to about 3000 pg / kg, about 1 pg / kg to about 2500 pg / kg, about 5 pg / kg to about Attorney Docket No.: S199207 1060WO (00093)

[0173] ST-020-W01

[0174] 2500 pg / kg, about 10 pg / kg to about 2500 pg / kg, from about 15 pg / kg to about 2500 pg / kg, from about 20 pg / kg to about 2500 pg / kg , from about 30 pg / kg to about 2500 pg / kg, about 1 pg / kg to about 2000 pg / kg, about 5 pg / kg to about 2000 pg / kg, about 10 pg / kg to about 2000 pg / kg, about 15 pg / kg to about 2000 pg / kg, from about 20 pg / kg to about 2000 pg / kg, from about 30 pg / kg to about 2000 pg / kg, about 1 pg / kg to about 1500 pg / kg, about 5 pg / kg to about 1500 pg / kg, about 10 pg / kg to about 1500 pg / kg, about 15 pg / kg to about 1500 pg / kg, from about 20 pg / kg to about 1500 pg / kg, from about 30 pg / kg to about 1500 pg / kg, about 1 pg / kg to about 1000 pg / kg, about 5 pg / kg to about 1000 pg / kg, about 10 pg / kg to about 1000 pg / kg, about 15 pg / kg to about 1000 pg / kg, from about 20 pg / kg to about 1000 pg / kg, from about 30 pg / kg to about 1000 pg / kg, about 1 pg / kg to about 500 pg / kg, about 5 pg / kg to about 500 pg / kg, about 10 pg / kg to about 500 pg / kg, about 15 pg / kg to about 500 pg / kg, from about 20 pg / kg to about 500 pg / kg, from about 30 pg / kg to about 500 pg / kg, about 1 pg / kg to about 250 pg / kg, about 5 pg / kg to about 250 pg / kg, about 10 pg / kg to about 250 pg / kg, about 15 pg / kg to about 250 pg / kg, from about 20 pg / kg to about 250 pg / kg, from about 30 pg / kg to about 250 pg / kg, etc.

[0175] In some instances, the DR-18 is administered at a dose of at least 1 pg / kg, at least 5 pg / kg, at least 10 pg / kg, at least 15 pg / kg, at least 20 pg / kg, at least 30 pg / kg, at least 60 pg / kg, at least 120 pg / kg, at least 240 pg / kg, at least 360 pg / kg, at least 480 pg / kg, at least 600 pg / kg, at least 720 pg / kg, at least 840 pg / kg, at least 960 pg / kg, at least 1080 pg / kg, or at least 1200 pg / kg or more.

[0176] In some instances, the DR-18 is administered at a dose of no more than 60 pg / kg, no more than 120 pg / kg, no more than 240 pg / kg, no more than 360 pg / kg, no more than 480 pg / kg, no more than 600 pg / kg, no more than 720 pg / kg, no more than 840 pg / kg, no more than 960 pg / kg, no more than 1080 pg / kg, or no more than 1200 pg / kg.

[0177] Where administered as multiple doses, the frequency of the administration of DR-18 can vary. For example, in some embodiments, each dose of successive doses of DR-18 is administered to the subject weekly, about every 7 days, at least 6 days after a previous dose of the composition, or at most 9 days after a previous dose. In some embodiments, each dose of successive doses of DR-18 is administered to the subject less frequently than weekly, including e.g., once every 2 weeks, once every three weeks, or once monthly.

[0178] In some instances, dosing may be done in cycles such that a first cycle is followed by at least a second cycle, including where the number of cycles may be 2, 3, 4, 2 or more, 3 or more, four or more, 4 or less, 3 or less, etc. Subsequent cycles may be the same or different. In some instances, a treatment regimen may include multiple cycles where all the cycles are the Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 same. In some instances, a treatment regimen may include multiple cycles where two or more of the cycles are different.

[0179] A dosing cycle may include any of the herein described dosing frequences. For example, in some embodiments, doses of DR-18 may be administered about every seven days in a cycle where, e.g., a cycle may include dosing on days 1 , 8, 15 and 22 of the cycle (or days 0, 7, 14 and 21 depending on the method used for counting). A treatment regimen may include a first cycle that includes dosing DR-18 on days 1 , 8, 15 and 22 of the first cycle and a second cycle that includes dosing DR-18 on days 8, 15 and 22 of the second cycle. In some embodiments, doses of DR-18 may be administered about every 14 days in a cycle where, e.g., a cycle may include dosing on days 1 and 15 the cycle. As described, multiples of the same or different cycles may be combined into a treatment regimen. For example, a treatment regimen may include dosing of DR-18 in a first cycle on days 1, 8, 15 and 22 of the first cycle, a second cycle on days 8, 15 and 22 of the second cycle, and a third cycle on days 1 and 15 of the third cycle. As in this case, dosing may become less frequent as cycles progress; however, in some instances, dosing may become more frequent as cycles progress or dosing frequency may stay the same. In some instances, a particular cycle, including any of those described herein, may be repeated indefinitely (e.g., until the subject reaches a clinical endpoint or event such as e.g., a complete response, disease progression, toxicity, or the like).

[0180] In some embodiments, a dose of the DR-18 composition comprises at least about 1 pg / kg, at least about 5 pg / kg, at least about 10 pg / kg, at least about 15 pg / kg, at least about 20 pg / kg, at least about 30 pg / kg, at least about 90 pg / kg, at least about 180 pg / kg, at least about 360 pg / kg, at least about 600 pg / kg, at least about 900 pg / kg, or at least about 1200 pg / kg of the polypeptide per body weight of the subject. In some embodiments, each dose of the engineered IL- 18 composition comprises between about 1 pg / kg to about 1200 pg / kg, between about 10 pg / kg to about 1200 pg / kg, between about 1 pg / kg to about 30 pg / kg, between about 10 pg / kg to about 30 pg / kg, between about 15 pg / kg to about 1200 pg / kg, between about 15 pg / kg to about 30 pg / kg, about 30 pg / kg to about 90 pg / kg, about 90 pg / kg to about 180 pg / kg, about 180 pg / kg to about 360 pg / kg, about 360 pg / kg to about 600 pg / kg, about 600 pg / kg to about 900 pg / kg, or about 900 pg / kg to about 1200 pg / kg of the polypeptide per body weight of the subject. In some embodiments, a dose of the engineered IL- 18 composition comprises about 1 pg / kg, about 10 pg / kg, about 15 pg / kg, about 30 pg / kg, about 60 pg / kg, about 90 pg / kg, about 120 pg / kg, about 180 pg / kg, about 240 pg / kg, about 360 pg / kg, about 600 pg / kg, about 900 pg / kg, or about 1200 pg / kg of the polypeptide per body weight of the subject. Attorney Docket No.: S199207 1060WO (00093)

[0181] ST-020-W01

[0182] In some embodiments, each dose of DR-18 comprises between about 60 pg / kg to about 1200 pg / kg, about 60 pg / kg to about 90 pg / kg, about 60 pg / kg to about 120 pg / kg, about 60 pg / kg to about 180 pg / kg, about 60 pg / kg to about 240 pg / kg, about 60 pg / kg to about 360 pg / kg, about 60 pg / kg to about 420 pg / kg, about 60 pg / kg to about 480 pg / kg, about 60 pg / kg to about 540 pg / kg, about 60 pg / kg to about 600 pg / kg, about 60 pg / kg to about 660 pg / kg, about 60 pg / kg to about 720 pg / kg, about 60 pg / kg to about 800 pg / kg, about 60 pg / kg to about 860 pg / kg, about 60 pg / kg to about 920 pg / kg, about 60 pg / kg to about 980 pg / kg, about 60 pg / kg to about 1040 pg / kg, or about 60 pg / kg to about 1200 pg / kg.

[0183] In some embodiments, each dose of DR-18 comprises between about 120 pg / kg to about 1200 pg / kg, about 120 pg / kg to about 180 pg / kg, about 120 pg / kg to about 240 pg / kg, about 120 pg / kg to about 360 pg / kg, about 120 pg / kg to about 420 pg / kg, about 120 pg / kg to about 480 pg / kg, about 120 pg / kg to about 540 pg / kg, about 120 pg / kg to about 600 pg / kg, about 120 pg / kg to about 660 pg / kg, about 120 pg / kg to about 720 pg / kg, about 120 pg / kg to about 800 pg / kg, about 120 pg / kg to about 860 pg / kg, about 120 pg / kg to about 920 pg / kg, about 120 pg / kg to about 980 pg / kg, about 120 pg / kg to about 1040 pg / kg, or about 120 pg / kg to about 1200 pg / kg.

[0184] Amounts administered and doses described herein can, in some instances reflect the amount used in initial dosing of DR-18 naive subjects, repeat dosing of previously dosed subject, or both. Accordingly, in some instances, the methods involve administering to subjects that have not been previously administered DR-18. In some embodiments, the methods involve administering DR-18 to subjects that have been previously administered the DR-18. In some embodiments, a dose described herein can be employed in initial dosing or repeat dosing. In some embodiments, repeat dosing can employ the same dose as used for initial dosing. In some embodiments, repeat dosing can employ one or more different doses as compared to what was employed for initial dosing.

[0185] In some embodiments, the dose of DR-18 employed in a method of the present disclosure where the subject is deficient in the production of anti-drug antibodies is a reduced dose, e.g., as in comparisons to a dose employed in a subject that is not deficient in the production of anti-drug antibodies. In some embodiments, the dose of DR-18 employed in a method of the present disclosure where the subject has a hematological cancer is a reduced dose, e g., as in comparison to a dose employed in a subject that has a cancer that is not a hematological cancer, such as e.g., a solid tumor. In some embodiments, the dose of DR-18 employed in a method of the present disclosure where the subject has a relapsed and / or refractory hematological cancer is a reduced dose, e.g., as in comparisons to a dose employed Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 in a subject that has a cancer that is not relapsed and / or refractory, such as a hematological cancer that is not relapsed and / or refractory.

[0186] In some embodiments, the dose of DR-18 administered to a subject with an impaired capacity to produce anti-drug antibodies (ADAs) and neutralizing antibodies (nAbs) can be a reduced dose as compared to the dose administered to a subject without such an impairment. This includes, for example, administering a dose that is at least 10% less than the standard dose employed for subjects without impaired ADA and nAb production. By way of further example, in some instances, the reduced dose may be at least 20% less, at least 30% less, at least 40% less, or at least 50% less than the dose administered to subjects without this impairment. For illustrative purposes, if a typical dose for a standard subject is 240 pg / kg, the reduced dose could be 216 pg / kg (10% less), 192 pg / kg (20% less), 168 pg / kg (30% less), 144 pg / kg (40% less), or 120 pg / kg (50% less) for a subject with a diminished capability for producing ADAs and nAbs.

[0187] In some embodiments, DR-18 (as a monotherapy or in combination with other therapeutics) is administered for at least 24 weeks. In some embodiments, DR-18 (as a monotherapy or in combination with other therapeutics) is administered until the subject has been administered at least 8 doses. In some embodiments, DR-18 (as a monotherapy or in combination with other therapeutics) is administered until the subject exhibits a complete response (CR), has been treated for at least 24 weeks, has been administered at least 8 doses, or some combination thereof. In some instances, DR-18 is administered (or administration is continued) regardless of subjects’ response or achievement of stable disease, including e.g., where such continued treatment is continued for months or years, including e.g., until the subject progresses.

[0188] In combination treatments, dosing of the combination agent(s) (i.e., the agent(s) other than DR-18) may be performed at the same frequency or a different frequency from the DR-18. For example, a DR-18 may be dosed weekly and a multi-specific therapeutic used in a combination therapy may also be dosed weekly. In some instances, a multi-specific therapeutic used in a combination therapy may be dosed less frequently than the DR-18. In some instances, a multi-specific therapeutic used in a combination therapy may be dosed more frequently than the DR-18. Useful dosing frequencies for a multi-specific therapeutic used in combination with DR-18 include but are not limited to e.g., twice daily, daily, every two days, every three days, every four days, every five days, every six days, twice weekly, weekly, every two weeks, every three weeks, twice monthly, monthly, and the like. In some instances, dosing of a multi-specific therapeutic may be done in cycles (as described herein for DR-18 dosing), Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 including e.g., where the multi-specific therapeutic is dosed about every two days within a cycle, about every seven days within a cycle, about every 14 days within a cycle, etc. As described for DR-18 dosing, dosing of a multi-specific therapeutic may increase or decrease in frequency over time or may stay the same. In some instances, previously used, or known, dosing of the multi-specific therapeutic, e.g., from prior clinical trials using the multi-specific therapeutic, may be employed.

[0189] In some embodiments of methods described herein, a Complete Blood Count (CBC) analysis may be performed, e.g., as a means of diagnosing and / or monitoring one or more of a disease, disease progression, therapy effectiveness, or the like. CBC is frequently used as a diagnostic test for detecting and monitoring blood cancers. This test measures various components of the blood, including red blood cells (RBC), white blood cells (WBC), hemoglobin, hematocrit, and platelets. Test results may be compared to baseline or reference parameters to identify abnormalities suggestive of blood cancers or quantitatively monitor disease.

[0190] In some embodiments of methods described herein, a flow cytometry analysis may be performed, e.g., as a means of diagnosing and / or monitoring one or more of a disease, disease progression, therapy effectiveness, or the like. Flow cytometry is a diagnostic tool employed to identify and characterize cell populations, including in blood cancers. This technique involves labeling cell surface markers with fluorescent antibodies and passing the cells through a laser- equipped cytometer. The emitted fluorescence is then measured to determine the presence and / or quantity of specific cell types. Flow cytometry is useful in diagnosing and monitoring different cancers, such as lymphomas and leukemias, as it can distinguish between different subtypes based on their surface markers. Flow cytometry provides detailed information about the cellular composition of a sample, allowing for the detection and characterization of disease and / or disease progression, including the detection of minimal residual disease and monitoring of treatment efficacy.

[0191] In some embodiments of methods described herein, a cytogenetic analysis may be performed, e.g., as a means of diagnosing and / or monitoring one or more of a disease, disease progression, therapy effectiveness, or the like. Cytogenetic analysis examines chromosomal abnormalities in cancer cells, including blood cancer cells, which analysis can provide diagnostic and prognostic information. Techniques such as karyotyping and fluorescence in situ hybridization (FISH) are commonly used in a cytogenetic analysis. Karyotyping involves arresting cells in metaphase, staining chromosomes, and analyzing their structure under a microscope to detect large-scale genetic alterations like translocations, deletions, and duplications. FISH uses fluorescent probes that bind to specific DNA sequences, allowing for Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 the visualization of genetic abnormalities at a molecular level. These cytogenetic techniques are frequently employed for identifying characteristic phenomena, such as e.g., Philadelphia chromosome translocations, and diagnosing various conditions, such as e.g., chronic myeloid leukemia (CML).

[0192] In some embodiments of methods described herein, a molecular diagnostic analysis may be performed, e.g., as a means of diagnosing and / or monitoring one or more of a disease, disease progression, therapy effectiveness, or the like. Molecular diagnostics involve analyzing specific genetic mutations and gene rearrangements associated with blood cancers. Techniques such as polymerase chain reaction (PCR) and next-generation sequencing (NGS) are widely used in molecular diagnostic analyses. PCR amplifies targeted DNA sequences to detect mutations or translocations characteristic of particular cancers, including blood cancers, such as the BCR-ABL fusion in CML. NGS provides a comprehensive analysis of the genetic landscape of a given sample, enabling the identification of multiple mutations and gene expressions at a high resolution. Molecular diagnostics aid in the early detection, classification, and personalized treatment planning of cancer treatment, including the treatment of blood cancers, e.g., by identifying actionable genetic alterations.

[0193] In some embodiments of methods described herein, a bone marrow aspiration or biopsy (BMAB) analysis may be performed, e.g., as a means of diagnosing and / or monitoring one or more of a disease, disease progression, therapy effectiveness, or the like. BMAB analyses involve invasive procedures used to diagnose and monitor blood cancers by directly sampling the bone marrow. During these procedures, a needle is inserted into the bone, usually the iliac crest, to extract liquid marrow (aspiration) or a core of marrow tissue (biopsy). The samples are then examined microscopically to evaluate cellular composition, architecture, and the presence of abnormal cells. Bone marrow analysis is frequently used for diagnosing, staging, and monitoring various cancers, including e.g., leukemias, myelomas, and lymphomas, as it provides information about the extent of disease infiltration, marrow function, and response to therapy.

[0194] In some embodiments of methods described herein, an imaging analysis may be performed, e.g., as a means of diagnosing and / or monitoring one or more of a disease, disease progression, therapy effectiveness, or the like. Imaging studies, including computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET) scans, play a vital role in diagnosing and monitoring various cancers, including blood cancers. These non- invasive techniques provide detailed cross-sectional images of the body's internal structures, e.g., allowing for the identification of lymph node involvement, organ infiltration, and tumor Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 masses. PET scans, often combined with CT (PET-CT), utilize radioactive tracers to highlight regions of increased metabolic activity, which may be indicative of cancerous cells. Imaging studies are frequently used for various purposes in cancer therapy, including blood cancer cancer therapy, including e.g., disease staging, assessing treatment response, and detecting disease recurrence or progression.

[0195] In some instances, analysis, e.g., on-treatment analysis or post-treatment analysis, can be compared to baseline analyses, such as e.g., baseline CBC, baseline flow cytometry, baseline cytogenetic analysis, baseline molecular diagnostics, baseline BMAB, baseline imaging, the like or combinations thereof. In some embodiments, methods of the present disclosure can include performing and / or collecting such baseline analyses and / or making comparisons between baseline analysis and one or more other analyses. For example, in some instances, imaging can be compared to baseline images, such as e.g., baseline scans, and the methods can include collecting such baseline images.

[0196] Comparisons may, correspondingly, be made between any analyses performed, including but not limited to e.g., comparisons between baseline and on-treatment analyses, comparisons between baseline and end-of-treatment analyses, comparisons between baseline and post-treatment analyses, comparisons between on-treatment analyses taken at different timepoints, comparisons between on-treatment analyses and end-of-treatment analyses, comparisons between on-treatment analyses and post-treatment analyses, comparisons between end-of-treatment analyses and post-treatment analyses, and the like.

[0197] In some instances, assessments can be made through the use of one or more biological samples, such as e.g., blood, plasma, bone marrow, and the like. In some instances, blood and plasma samples can be collected and used for pharmacokinetic (PK) and pharmacodynamic (PD) analysis, as well as for measuring other characteristics, such as presence / absence / amount of cancerous cells, presence / absence / amount of healthy immune cells, presence / absence / amount of ADA, presence / absence / amount of nAbs, and / or identifying select biomarkers.

[0198] In some embodiments, methods described herein include administering an additional agent that enhances the therapeutic efficacy of the DR-18 treatment. Put another way, in some instances, methods of the present disclosure include administering a combination therapy that includes a plurality of different agents where the DR-18 represents one agent of the plurality. Results achieved with the combination therapy are superior to the results achieved by each of the individual agents administered separately as monotherapies. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0199] Such additional agents include, e.g., antibodies that target an antigen on cancer cells present in the subject. Useful antibodies may vary and may include, e.g., monoclonal antibodies, monospecific antibodies, bispecific antibodies, trispecific antibodies, and the like. In some embodiments, such a combination therapy is administered to a subject having a hematological cancer (such as e.g., a leukemia, a lymphoma, a myeloma, etc.), including relapsing and / or refractory hematological cancers, including e.g., a relapsing and / or refractory leukemia, a relapsing and / or refractory lymphoma, a relapsing and / or refractory myeloma, etc. In some embodiments, such a combination therapy is administered to a subject having a solid tumor, including a solid tumor that is a primary tumor, a relapsed or recurrent solid tumor, a refractory solid tumor, a resistant solid tumor, a metastatic solid tumor, a locally advanced solid tumor, or any combination thereof.

[0200] In some instances, the additional agent is a monoclonal antibody. Useful monoclonal antibodies include those with an antigen-binding domain that specifically binds to an antigen present on the cancer cells within the subject. Useful antigens on cancer cells to which an antibody will bind, described in more detail below, will vary and may include essentially any antigen differentially expressed on cancer cells to be targeted. For example, in some instances, useful antigens may include immune checkpoint antigens expressed on cancer cells (i.e. , cancer cell antigens targeted by immune checkpoint inhibitors). In other instances, useful antigens may include cancer antigens other than immune checkpoint antigens (i.e., cancer cell antigens that are not immune checkpoint antigens). Put another way, in some instances, an antigen on a cancer targeted by an antibody employed in a method of the present disclosure is not an immune checkpoint antigen, such as e.g., PD-1 (Programmed Cell Death Protein 1), PD- L1 (Programmed Death-Ligand 1), PD-L2 (Programmed Death-Ligand 2), CTLA-4 (Cytotoxic T- Lymphocyte Associated Protein 4), TIM-3 (T-cell Immunoglobulin and Mucin-domain containing- 3), TIGIT (T cell Immunoreceptor with Ig and ITIM domains), or LAG-3 (Lymphocyte Activation Gene-3), as such antigens may be expressed on cancer cells or immune cells and targeted using checkpoint inhibitors. In some instances, an antigen will be differentially expressed on a cancer cell by at least 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50- fold, or 100-fold or more as compared to a comparable non-cancerous cell.

[0201] In some embodiments, useful antibodies, including useful monoclonal antibodies, include an anti-CD20 antibody, which selectively targets CD20-expressing cells. For example, in some embodiments, the method involves evaluating, or having evaluated, the subject's immunological profile and determining the therapeutic need for treatment with the anti-CD20 antibody to enhance the cytotoxic activity against hematological malignancies. CD20 binding Attorney Docket No.: S199207 1060WO (00093)

[0202] ST-020-W01 antibodies (and the CD20 antigen binding domains thereof) are known to the relevant skilled artisan and include, e.g., antibodies including the heavy and light chains of divozilimab, ibritumomab, obinutuzumab, ocaratuzumab, ocrelizumab, ofatumumab, ripertamab, rituximab, ublituximab, veltuzumab, zuberitamab, and the like. Useful antigen binding domains include those derived from the heavy chain, light chain, and / or CDRs of antibodies that bind to CD20, such as but not limited to e.g., divozilimab, ibritumomab, obinutuzumab, ocaratuzumab, ocrelizumab, ofatumumab, ripertamab, rituximab, ublituximab, veltuzumab, zuberitamab, and the like.

[0203] In some embodiments, the method includes administering an anti-CD38 antibody. CD38 antibodies specifically binds to the CD38 antigen, e.g., as expressed on multiple myeloma cells. The administration of the anti-CD38 antibody, in conjunction with DR-18, may target refractory multiple myeloma cells, leveraging the antibody's cytotoxic mechanisms to improve treatment outcomes. CD38 binding antibodies (and the antigen binding domains thereof) are known to the relevant skilled artisan and include, e.g., the heavy and light chains of daratumumab, erzotabart, felzartamab, isatuximab, lumrotatug, mezagitamab, modakafusp, and the like. Useful antigen binding domains include those derived from the heavy chain, light chain, and / or CDRs of antibodies that bind to CD38, such as but not limited to e.g., daratumumab, erzotabart, felzartamab, isatuximab, lumrotatug, mezagitamab, modakafusp, and the like.

[0204] In some embodiments, the additional agent (administered with the DR-18) may be a bispecific antibody, such as e.g., a bispecific antibody targeting CD19 and CD3. Such a bispecific antibody is capable of facilitating the recruitment and activation of T cells against CD19-positive B cell malignancies. One example of a useful bispecific antibody targeting CD19 and CD3 is blinatumomab.

[0205] CD19 antibodies (and the antigen binding domains thereof that may be employed in a monospecific or multi-specific antibody format) are known to the relevant skilled artisan and include, e.g., the heavy and light chain domains of budoprutug, coltuximab, denintuzumab, inebilizumab, leronlimab, loncastuximab, mogamulizumab, obexelimab, plozalizumab, tafasitamab, and the like. Useful antigen binding domains include those derived from the heavy chain, light chain, and / or CDRs of antibodies that bind to CD19, such as but not limited to e.g., budoprutug, coltuximab, denintuzumab, inebilizumab, leronlimab, loncastuximab, mogamulizumab, obexelimab, plozalizumab, tafasitamab, and the like.

[0206] In some embodiments, the methods of the present disclosure include selecting subjects with reduced ADA and nAb responses and administering one or more additional agents, such as an antibody, multi-specific antibody, bispecific antibody, etc., following DR-18 treatment, Attorney Docket No.: S199207 1060WO (00093)

[0207] ST-020-W01 thereby promoting enhanced immune-mediated destruction of cancer cells. In some embodiments, the methods of the present disclosure include selecting subjects with reduced ADA and nAb responses and administering one or more additional agents, such as an antibody, multi-specific antibody, bispecific antibody, etc., during DR-18 treatment, thereby promoting enhanced immune-mediated destruction of cancer cells. In some embodiments, the methods of the present disclosure include selecting subjects with reduced ADA and nAb responses and administering one or more additional agents, such as an antibody, multi-specific antibody, bispecific antibody, etc., before DR-18 treatment, thereby promoting enhanced immune- mediated destruction of cancer cells.

[0208] In some embodiments, the method involves the use of an anti-BCMA antibody, which binds to B cell maturation antigen (BCMA), including e.g., where the BCMA antigen is expressed on multiple myeloma cells. In some embodiments, the antibody is a BCMA bispecific antibody that binds BCMA and an antigen expressed on effector T cells present in the subject. Such a BCMA bispecific antibody may engage BCMA on malignant plasma cells and simultaneously bind to an antigen, such as CD3, which is expressed on effector T cells. In doing so, the BCMA bispecific antibody may facilitate an immunological synapse between the malignant plasma cells and effector T cells, potentially leading to targeted cell lysis.

[0209] Examples of antibodies, including monospecific and multi-specific antibodies, targeting BCMA include but are not limited to e.g., podentamig, alnuctamab, belantamab, cizutamig, elranatamab, etentamig, ingitamig, ispectamab, linvoseltamab, pacanalotamab, pamlectabart, pavurutamab, teclistamab, velinotamig, and vonsetamig.

[0210] In some embodiments, multi-specific antibodies, such as bispecific antibodies, may include variable regions that specifically target a cancer antigen (e.g., CD19, CD20, BCMA, etc.) and the chosen antigen on effector T cells (e.g., CD3). These variable regions may be derived from any available and appropriate method, including e.g., hybridoma technology, phage display, or humanization techniques, among other methods. Where employed, a bispecific antibody may also incorporate various formats such as tandem single-chain variable fragments (scFvs), dual variable domain antibodies (DVD-lg), or other engineered bispecific constructs known in the art, which enable binding to both the cancer antigen (e.g., CD19, CD20, BCMA, etc.) and the antigen on effector T cells (e.g., CD3).

[0211] In some instances, a combination of DR-18 and an antibody, such as a monoclonal antibody, targeting CD33 may be employed. Such a method may involve administering the anti- CD33 antibody in subjects with acute myeloid leukemia (AML), enhancing the specific targeting of CD33-positive leukemic cells and improving overall treatment efficacy. CD33 antibodies (and Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 antigen binding domains thereof useful in monospecific and multi-specific antibody formats) are known to the relevant skilled artisan and include, e.g., the heavy and light chains of gemtuzumab, lintuzumab, vadastuximab, and the like. Useful antigen binding domains include those derived from the heavy chain, light chain, and / or CDRs of antibodies that bind to CD33, such as but not limited to e.g., gemtuzumab, lintuzumab, vadastuximab, and the like.

[0212] Examples of bispecific antibodies Targeting CD33 include but are not limited to e.g., AMG 330, AMV564, and JNJ-67571244. CD33-targeting bispecific antibodies harness the immune system's potential to target CD33+ cancer cells via various mechanisms of action involving T cell or NK cell recruitment and activation.

[0213] Methods of the present disclosure are employed to treat cancer. In some embodiments, methods of the present disclosure treat solid tumors. In some embodiments, methods of the present disclosure treat blood cancers (also referred to as hematological malignancies). In some instances, i.e. , where the methods are employed to treat hematological malignancies, the treated subjects are not subjects with solid tumors.

[0214] The following are non-limiting examples of cancers that can be treated (or re-treated in the case of cancer relapse) or prevented (preventing relapse) by the methods and compositions of the disclosure: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, appendix cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain and spinal cord tumors, brain stem glioma, brain tumor, breast cancer, bronchial tumors, Burkitt lymphoma, carcinoid tumor, central nervous system atypical teratoid / rhabdoid tumor, central nervous system embryonal tumors, central nervous system lymphoma, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, cerebral astrocytoma / malignant glioma, cervical cancer, childhood visual pathway tumor, chordoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous cancer, cutaneous t-cell lymphoma, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing family of tumors, extracranial cancer, extragonadal germ cell tumor, extrahepatic bile duct cancer, extrahepatic cancer, eye cancer, fungoides, gallbladder cancer, gastric (stomach) cancer, gastrointestinal cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (gist), germ cell tumor, gestational cancer, gestational trophoblastic tumor, glioblastoma, glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic and visual pathway glioma, hypothalamic tumor, intraocular (eye) cancer, intraocular melanoma, islet cell tumors, Kaposi sarcoma, kidney (renal cell) cancer, Langerhans cell cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, Attorney Docket No.: S199207 1060WO (00093)

[0215] ST-020-W01 lip and oral cavity cancer, liver cancer, lung cancer, lymphoma, macroglobulinemia, malignant fibrous histiocytoma of bone and osteosarcoma, medulloblastoma, medulloepithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, mouth cancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosis, myelodysplastic syndromes, myelodysplastic / myeloproliferative diseases, myelogenous leukemia, myeloid leukemia, myeloma, myeloproliferative disorders, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oral cavity cancer, oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, papillomatosis, paraganglioma, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal parenchymal tumors of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumor, plasma cell neoplasm, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, primary central nervous system cancer, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter cancer, respiratory tract carcinoma involving the nut gene on chromosome 15, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, sezary syndrome, skin cancer (melanoma), skin cancer (nonmelanoma), skin carcinoma, small cell lung cancer, small intestine cancer, soft tissue cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer, stomach (gastric) cancer, supratentorial primitive neuroectodermal tumors, supratentorial primitive neuroectodermal tumors and pineoblastoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell cancer, transitional cell cancer of the renal pelvis and ureter, trophoblastic tumor, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma, vulvar cancer, waldenstrom macroglobulinemia, and Wilms Tumor.

[0216] In some non-limiting examples, the cancer is defined not (or at least not exclusively) by the histological tissue of origin, but by its molecular features. For example, tumors with high tumor mutational burden (TMB), tumors with microsatellite instability (MSI), tumors that lack or have diminished surface expression of MHC Class I (e.g., due to deletion of Beta-2 microglobulin or Tapasin), or tumors (or patients or samples thereof) that express high levels of IL-18BP. In some cases, those molecular features are determined through next-generation sequencing (e.g., using TMB / MSI). In some cases, those molecular features are determined through immunohistochemistry, immunofluorescence, or flow cytometry. Thus, non-limiting Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 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, including any of the foregoing with a high TMB, high MSI, no or low MHC class I, high IL-18BP, or any combination thereof.

[0217] In some instances, the methods include treating a subject for a liquid tumor, e.g., a blood cancer. Accordingly, in some instances, the cancer for which the subject is treated is not a solid tumor. In some instances, the blood cancer is a lymphocyte neoplasm. In some instances, the blood cancer is a B-cell neoplasm. In some instances, the blood cancer is a T-cell neoplasm. In some instances, the blood cancer is a plasma cell neoplasm. In some instances, the blood cancer is a lymphoma. In some instances, the blood cancer is a leukemia. In some instances, the blood cancer is of neutrophil, mast cell, macrophage, or dendritic cell origin. In some instances, the methods include treating a subject for a solid tumor. In some instances, the cancer for which the subject is treated is not a liquid cancer, not blood cancer, not a B-cell neoplasm, not a T-cell neoplasm, not a plasma cell neoplasm, not a lymphoma, not nonHodgkin's lymphoma, not a leukemia, or the like.

[0218] In some embodiments, the methods of the present disclosure are useful for treating a tumor or cancer that is resistant to immune checkpoint inhibitors (ICIs). Examples of immune checkpoint inhibitors include, but are not limited to, anti-PD1 agents such as an anti-PD1 antibody (e.g., pembrolizumab, zimberelimab, nivolumab, cemiplimab, dostarlimab), anti-PD-L1 agents such as an anti-PD-L1 antibody (e.g., avelumab, durvalumab, atezolizumab), anti-PD-L2 agents such as an anti-PD-L2 antibody, anti-CTLA4 (e.g., ipilimumab, tremelimumab), anti- TIM3, anti-TIGIT, anti-LAG3 (e.g., relatlimab), anti-B7H3 (e.g., enoblituzumab), anti-B7H4, anti- VISTA, anti-BTLA, anti-CD47, anti-SIRP alpha, anti-CD48, anti-CD155, anti-CD160, anti- TREM2, anti-l DO1 , anti-Adenosine 2A receptor, anti-Aryl hydrocarbon receptor, anti-KIR, and anti-LILRB2. Examples of targets of immune checkpoint inhibitors include but are not limited to PD-L1 , PD-L2, PD1 , CTLA4, TIM3, TIGIT, LAG3, B7-1 , B7H3, B7H4, VISTA, BTLA, CD28, CD47, SIRP alpha, CD48, CD155, CD160, TREM2, IDO1 , Adenosine 2A receptor, Aryl hydrocarbon receptor, KIR, LILRB2, and combinations thereof. Other examples of ICIs and targets of ICIs to which a subject’s cancer may be resistant are described elsewhere herein.

[0219] In some embodiments, methods of the present disclosure may include administering cytokine release syndrome (CRS) prophylaxis to the subject. CRS prophylaxis, or a CRS prophylactic agent, can be administered to a subject starting from the first dose of DR-18. In some embodiments, the CRS prophylactic agent is administered as premedication or post-dose Attorney Docket No.: S199207 1060WO (00093)

[0220] ST-020-W01 medication to a dose of DR- 18. In some embodiments, the method further comprises administering a dose of a CRS prophylactic agent to the subject with or before a dose or successive doses of DR-18. In some embodiments, the method comprises administering a dose of a CRS prophylactic agent to the subject with or before each dose of the successive doses of the DR IL-18 composition. Useful CRS prophylactic agents may include a non-steroidal antiinflammatory drug (NSAID), acetaminophen, diphenhydramine, histamine H1 antagonist, famotidine, H2 blocker, or fluids. In some embodiments, CRS prophylaxis may include administering a non-steroidal anti-inflammatory drug (NSAID), acetaminophen, diphenhydramine, histamine H1 antagonist, famotidine, H2 blocker, fluids, or some combination thereof. Useful CRS prophylactic agents, CRS prophylaxis regimens, and information related thereto (including e.g., administration timing and dosages, are described in WO2024 / 158838, the disclosure of which is incorporated herein by reference in its entirety.

[0221] In some embodiments, the method comprises administering to a subject, or contacting a cell or tissue with, an isolated nucleic acid molecule encoding a DR-18 polypeptide. In some embodiments, a DR-18 polypeptide is administered to a cell, tissue, organ, system, or subject to treat a disease or disorder, including e.g., where such a DR-18 polypeptide is administered as a nucleic acid (e.g., DNA, cDNA, mRNA, etc.) encoding the DR-18 polypeptide. DR-18 encoding nucleic acids may be administered by any convenient and appropriate means, such as "naked" nucleic acid (e.g., naked DNA, mRNA, etc.), via a viral vector (e.g., adenovirus, adeno- associated virus, lentivirus, etc.), via a non-viral vector (e.g., a lipid nanoparticle), or within a cell (e.g., genome integrated or extrachromosomally). As will be readily understood by a relevant artisan, references herein to use of a DR-18 polypeptide in the methods of the present disclosure may be substituted and adapted for use with a nucleic acid molecule encoding the DR-18 polypeptide.

[0222] During and / or after performance of the treatment method as described herein, one or more outcomes may be measured. Useful outcome measures include but are not limited to e.g., overall response rate (ORR), duration of response (DOR), progression-free survival (PFS), overall survival (OS), minimal / measurable residual disease (MRD) negativity, complete response (OR), and partial response (PR). In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the ORR as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, or 300% or more increase in the DOR as compared to a Attorney Docket No.: S199207 1060WO (00093)

[0223] ST-020-W01 corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the PFS period as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the OS rate as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the occurrence of MRD negativity as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the duration of MRD negativity as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the CR rate as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the duration of CR as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the PR rate as compared to a corresponding protocol not employing DR-18 and / or SOC therapy. In some instances, employing DR-18 in a method of treatment as described herein, results in an at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more increase in the duration of PR as compared to a corresponding protocol not employing DR-18 and / or SOC therapy.

[0224] Any convenient and appropriate method for measuring ORR, DOR, PFS, and / or OS may be employed, including e.g., where Kaplan-Meier methodology is used to assess ORR, DOR, PFS, and OS. In some instances, ORR will be defined as a composite of CR plus PR. In some instances, measures of ORR, DOR, PFS, OS, CR, PR, and the like may be assessed according to established response criteria for a particular condition. For example, in some instances, response criteria, such as for ORR, CR, or the like, will be determined for AML or MDS using the European LeukemiaNet 2022 response criteria (see e.g., Dbhner et al. Blood (2022) 140 (12): 1345-1377). In some instances, criteria may be combined, including e.g., Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 where PR for AML or MDS will be defined as 25% reduction in blasts in subjects and response assessed according to European LeukemiaNet 2022 criteria.

[0225] As noted above, in some instances, response criteria for a particular condition may be employed. For example, for BCL, CR and ORR may be assessed according to the Lungo classification (see e.g., Cheson et al. J Clin Oncol (2014) 32(27): 3059-68). For multiple myeloma, ORR may be assessed according to the International Myeloma Working Group (IMWG) criteria (see e.g., Rajkumar et al. Lancet Oncol (2014) 15(12):e538-48).

[0226] MRD negativity may be assessed by any convenient and appropriate method including e.g., using qPCR (qPCR-MRD) (e.g., of a bone marrow or peripheral blood sample), inc. e.g., molecular MRD (Mol-MRD) assessed by qPCR, flow cytometry (e.g., of a bone marrow sample or other relevant tissue sample), inc. e.g., multiparameter flow cytometry MRD (MFC-MRD), next-generation sequencing (NGS), and digital PCR (dPCR). See e.g., Heuser et al. Blood. (2021) 138(26): 2753-2767. MRD positivity / negativity is useful in assessing various aspects of various hematological cancers, including but not limited to e.g., the presence, absence, prognosis, risk of relapse, effectiveness of treatment, etc. of a hematological cancer, a lymphoma, a leukemia, a myeloma, ALL, AML, MM, and the like.

[0227] Over time clinicians have promoted the replacement of “minimal” residual disease with “measurable” residual disease as the preferred definition of MRD to reflect the shift to assessing MRD based on “positive” or “negative” detection of measurable disease according to specific thresholds. Accordingly, a negative MRD result (or “MRD negativity”) does not necessarily indicate disease eradication but, rather, represents disease below the assay’s threshold in the tested sample and an assay with a non-zero result may still be labeled “negative” by a laboratory if the level detected is below the threshold linked to prognosis. It is noted that any description of a “negativity” threshold will be readily understood as also providing the corresponding “positivity” threshold.

[0228] In some instances, MRD is assessed using flow cytometry, such as MFC, and MFC- MRD test positivity is defined as greater than or equal to 0.1% of CD45-expressing cells with the target immunophenotype (such that MRD negativity is defined as less than 0.1 %). In some instances, the positivity (and correspondingly the negativity) threshold is lower than 0.1%, including e.g., where MRD negativity is defined as less than 0.095%, 0.090%, 0.085%, 0.080%, 0.075%, 0.070%, 0.065%, 0.060%, 0.055%, 0.050%, 0.045%, 0.040%, 0.035%. 0.030%, 0.025%, 0.020%, 0.015%, 0.010%, or less of CD45-expressing cells with the target immunophenotype. In some instances, MRD positivity is defined any detectable MRD by flow cytometry, and MRD negativity is correspondingly defined as no detectable CD45-expressing Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 cells with the target immunophenotype by flow cytometry according to the sensitivity threshold of the assay.

[0229] In some instances, MRD is assessed using flow cytometry through detection of malignant blasts, including where MRD positivity / negativity is based on detection above / below a threshold. For example, in some instances, MRD test positivity is defined as greater than or equal to 5% malignant blasts (such that MRD negativity is defined as less than 5% malignant blasts). In some instances, the positivity (and correspondingly the negativity) threshold is lower than 5%, including e.g., where MRD negativity is defined as less than 4.5%, 4.0%, 3.5%, 3.0%, 2.5%, 2.0%, 1.5%, 1.0%, or less malignant blasts. In some instances, MRD positivity is defined any detectable MRD by flow cytometry, and MRD negativity is correspondingly defined as no detectable malignant blasts by flow cytometry according to the sensitivity threshold of the assay.

[0230] In some instances, MRD is assessed using qPCR. In some instances, qPCR is preferred where one or more markers of disease or targetable disease abnormalities (e.g., mutations and / or fusion transcripts) have been detected in the subject’s disease. Useful markers of disease and for measuring MRD in AML include e.g., gene mutations associated with disease and / or used in defining of molecularly defined disease subgroups and / or known as disease abnormalities, such as but not limited to, e.g., AML mutations in NPM1 , RUNX1-RUNX1T1 , CBFB-MYH11 , PML-RARA, KMT2A-MLLT3, DEK-NUP214, BCR-ABL, and WT1. Useful samples for qPCR MRD assessments include peripheral blood and bone marrow. Different markers may be employed for assessing MDR by qPCR in different diseases. For example, useful markers of disease and for measuring MRD in multiple myeloma include e.g., immunoglobulin heavy chain gene rearrangements, lambda and kappa light chain rearrangements, translocations such as t(4;14) and t(11 ;14), and other chromosome abnormalities. Methods for assessing MRD in BCL include but are not limited to those described in Merryman et al. Blood Adv (2023) 7 (17): 4748-4759; Atout et al. Clin Lymphoma Myeloma Leuk (2025) S2152-2650(25)00154-5.

[0231] In some instances, MRD is assessed using qPCR and MRD positivity / negativity is based on detection above / below a threshold, such as a cycling threshold. A cycling threshold (a.k.a. “cycle threshold”, “threshold cycles”, “Ct”, and “Cq”) represents the number of cycles needed to replicate enough of a template nucleic acid during a PCR reaction to detect the product of the reaction above a predefined threshold. A lower Ct value means more template nucleic acid was present at the start of the reaction (because fewer cycles were needed to reach the threshold) and a higher Ct value means less template nucleic acid was present at the start of the reaction (because more cycles were needed to reach the threshold). Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0232] In some instances, MRD is assessed using qPRC and MRD positivity is based on a cycling threshold of less than 40 cycles and accordingly, MRD negativity may be based on a cycling threshold of equal to or greater than 40 cycles. In some instances, such a MRD positivity / negativity cycling threshold of 40 cycles may be used in assessing MRD positivity / negativity for AML. Useful cycling thresholds may vary, e.g., depending on the cancer being assessed, the qPCR marker(s) being evaluated, the reason for the MRD assessment, and the like. In some instances, the positivity (and correspondingly the negativity) threshold is less than 40 cycles, including e.g., 39 cycles, 38 cycles, 37 cycles, 36 cycles, 35 cycles, 34 cycles, 33 cycles, 32 cycles, 31 cycles, or 30 cycles, or less. In some instances, the positivity (and correspondingly the negativity) threshold is more than 40 cycles, including e.g., 41 cycles, 42 cycles, 43 cycles, 44 cycles, or 45 cycles, or more.

[0233] Markers and methods employed in qPCR may be correspondingly employed for MDR assessments by other molecular-based or nucleic acid based methods, such as sequencing based methods such as, e.g., NGS-based MRD assessment. NGS-based MRD assessment thresholds may reference the assay’s sensitivity for cancer cells within a population of normal cells. For example, in some instances, achievement of MRD negativity (e.g., for multiple myeloma) may be based on a threshold by NGS of 10A-5, where A 10A-5 threshold signifies that the test can detect a minimum of one cancer cell within every 100,000 normal cells. In some instances, a sensitivity greater than 10A-5 may be employed, including e.g., where the MRD negativity threshold employed is 10A-6, which indicates the assay can detect one cancer cell in 1 ,000,000 normal cells. Any convenient and appropriate sample may be assessed, e.g., in some instances, an assessment for MRD in multiple myeloma may employ a bone marrow sample and may, e.g., use an MRD threshold of 10A-5.

[0234] In some instances, a subject treated according to the methods as described herein may achieve MRD negativity and, e.g., may be more likely to achieve MRD negativity as compared to a subject treated without administration of DR-18. In some instances, a subject treated according to the methods as described herein may achieve sustained MRD negativity, including e.g., where MRD negativity obtained by such a subject is more likely and / or more sustained as compared to a subject treated without administration of DR-18. A subject treated with the methods described herein may obtain sustained MRD (as assessed by multiple rounds of MRD assessment) for one month or greater, including at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or more.

[0235] Lack of sustained MRD negativity may, in some instances, be referred to as MRD relapse. The likely time period and characteristics of MRD relapse may vary for different Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 conditions. In some instances, MRD relapse may be defined as conversion of MRD negativity to MRD positivity in successive tests, and / or an increase in measured MRD in one or more tested samples. In some embodiments, treatment of subject according to the methods described herein may prevent MRD relapse. In some embodiments, subjects treated according to the methods described herein may experience a lower rate of MRD relapse as compared to the rate of MRD relapse in corresponding subjects treated without DR-18.

[0236] Further useful description of MRD assessments may be found in Heuser et al. Blood (2021) 138 (26): 2753-2767 and Dbhner et al. Blood (2022) 140 (12): 1345-1377.

[0237] Methods of the present disclosure provide advantages as compared to other methods. For example, methods combining DR-18 with one or more additional therapies, such as e.g., a combination therapy with a multi-specific therapeutic, provide advantages as compared to corresponding methods employing monotherapy treatment with either DR-18 alone or the multispecific therapeutic. Methods employing patient selection provide advantages as compared to corresponding methods not employing patient selection. Such advantages may manifest in one or more improved metrics, where any convenient and appropriate metric, or combination thereof, may be employed including but not limited to e.g., MRD, ORR, DOR, PFS, OS, CR, PR, and the like.

[0238] Agents & Compositions

[0239] The methods, and the kits that may be employed in performing such methods, described herein make use of certain agents, and compositions comprising such agents. Useful agents in the herein described methods include, e.g., DR-18 polypeptides, DR-18 encoding nucleic acids, monospecific antibodies, multi-specific therapeutics, such as bispecific, trispecific, and tetraspecific antibodies, and the like.

[0240] Decoy-resistant IL-18 (DR-18) polypeptides bind to, and signal through formation of, the IL-18Ra (IL-18 receptor a) and IL-18R|3 (IL-18 receptor P) receptor complex. Decoy- resista nt IL- 18 polypeptides do not bind to IL-18 binding protein (IL-18BP), or at least display substantially reduced binding to IL-18BP, such as substantially reduced binding to IL-18BP relative to wildtype (WT) human IL-18 (SEQ ID NO: 1) (i.e., as compared to the binding of IL-18BP to WT IL- 18). In some embodiments, the DR-18 polypeptide does not bind to IL-18BP. In some embodiments, the DR-18 polypeptide has reduced binding to IL-18BP relative to WT IL-18. In some embodiments, the DR-18 polypeptide binds to IL-18Ra and does not bind to IL-18BP. In some embodiments, the DR IL-18 polypeptide binds to IL-18Ra and has reduced binding to IL- 18BP relative to WT IL- 18. Attorney Docket No.: S199207 1060WO (00093)

[0241] ST-020-W01

[0242] WT human IL-18: YFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFIISMYKDSQPRGMAVTIS VKCEKISTLSCENKIISFKEMNPPDNIKDTKSDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKE RDLFKLILKKEDELGDRSIMFTVQNED (SEQ ID NO: 1)

[0243] WT human IL-18 is expressed initially in a pre-processed (unprocessed) form (SEQ ID NO: 65) that includes a 36 amino acid propeptide (SEQ ID NO: 66). DR-18 encoding nucleic acids may include or exclude sequence encoding the WT human IL-18 propeptide (SEQ ID NO: 66). IL-18 polypeptides expressed with a WT human IL-18 propeptide are subsequently processed to produce the active mature form of the polypeptide. Unless clearly intended otherwise, references to WT IL-18 polypeptides and DR-18 polypeptides will generally refer to the active mature forms of the polypeptides (i.e., without a WT human IL-18 propeptide). For example, reference to a DR-18 polypeptide having X mutations relative to WT human IL-18 shall, unless specifically indicated otherwise, refer to X mutations occurring within amino acids 1 to 157 of SEQ ID NO: 1 or amino acids 37 to 193 of SEQ ID NO: 65.

[0244] WT human IL-18 (unprocessed form): MAAEPVEDNCINFVAMKFIDNTLYFIAEDDENLESDYFGKLESKLSVIRNLNDQVLFIDQGNRPL FEDMTDSDCRDNAPRTIFIISMYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDT KSDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELGDRSIMFTVQNED (SEQ ID NO:65)

[0245] WT human IL-18 propeptide: MAAEPVEDNCINFVAMKFIDNTLYFIAEDDENLESD (SEQ ID NO: 66)

[0246] In some embodiments, the DR-18 polypeptide has at least about two mutations, at least about three mutations, at least about four mutations, at least about five mutations, at least about six mutations, at least about seven mutations, at least about eight mutations, at least about nine mutations, or at least about ten mutations, relative to WT IL-18 as set forth in SEQ ID NO: 1. In some embodiments, the DR-18 polypeptide has about two mutations, about three mutations, about four mutations, about five mutations, about six mutations, about seven mutations, about eight mutations, about nine mutations, or about ten mutations, relative to WT IL-18 as set forth in SEQ ID NO: 1. In some instances, a DR-18 polypeptide may comprise an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with WT IL-18 as set forth in SEQ ID NO: 1 .

[0247] Useful DR-18 polypeptides include but are not limited to e.g., a DR-18 polypeptide comprising any of the amino acid sequences of SEQ ID NOs.:2-63. In some instances, a DR-18 polypeptide may comprise an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0248] 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with at least one of SEQ ID NOs: 2-63. Useful DR-18 polypeptides include but are not limited to e.g., a DR-18 polypeptide comprising any of the amino acid sequences of SEQ ID NOs.:7, 17-22, 55-57, 62 and 63. In some instances, a DR-18 polypeptide may comprise an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with at least one of SEQ ID NOs: 7, 17-22, 55-57, 62 and 63.

[0249] In some instances, a useful DR-18 polypeptide comprises at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, or 15 mutated residues relative to SEQ ID NO:1. In some instances, a useful DR- 18 polypeptide comprises up to 15, 14, 13, 12, 11 , or 10 mutated residues relative to SEQ ID NO:1. In some instances, a useful DR-18 polypeptide comprises from 1 to 20, from 2 to 20, from 3 to 20, from 4 to 20, from 5 to 20, from 6 to 20, from 7 to 20, from 8 to 20, from 9 to 20, from 10 to 20, from 11 to 20, from 12 to 20, from 13 to 20, from 14 to 20, from 15 to 20, from 1 to 18, from 2 to 18, from 3 to 18, from 4 to 18, from 5 to 18, from 6 to 18, from 7 to 18, from 8 to 18, from 9 to 18, from 10 to 18, from 11 to 18, from 12 to 18, from 13 to 18, from 14 to 18, from 15 to 18, from 1 to 16, from 2 to 16, from 3 to 16, from 4 to 16, from 5 to 16, from 6 to 16, from 7 to 16, from 8 to 16, from 9 to 16, from 10 to 16, from 11 to 16, from 12 to 16, from 13 to 16, from 14 to 16, from 1 to 14, from 2 to 14, from 3 to 14, from 4 to 14, from 5 to 14, from 6 to 14, from 7 to 14, from 8 to 14, from 9 to 14, from 10 to 14, from 11 to 14, from 12 to 14, from 1 to 12, from 2 to 12, from 3 to 12, from 4 to 12, from 5 to 12, from 6 to 12, from 7 to 12, from 8 to 12, from 9 to 12, from 10 to 12, from 1 to 10, from 2 to 10, from 3 to 10, from 4 to 10, from 5 to 10, from 6 to 10, from 7 to 10, or from 8 to 10 mutated residues relative to SEQ ID NO:1.

[0250] In some instances, a useful DR- 18 polypeptide comprises an amino acid sequence of any one of SEQ I D NOs: 2-63 with 0, 1 , 2, 3, 4, 5, or 6 additional mutated residues relative to SEQ ID NO:1. In some instances, a useful DR-18 polypeptide comprises an amino acid sequence of any one of SEQ ID NOs: 7, 17-22, 55-57, 62 and 63 with 0, 1 , 2, 3, 4, 5, or 6 additional mutated residues relative to SEQ ID NO:1. In some instances, a useful DR-18 polypeptide comprises an amino acid sequence of any one of SEQ ID NOs: 7, 17-22, 55-57, 62 and 63 with up to 2, up to 3, up to 4, up to 5, or up to 6 additional mutated residues relative to SEQ ID NO:1.

[0251] In some instances, a useful DR- 18 polypeptide comprises one or more mutations at positions Y1 , L5, K8, C38, M51 , K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, D110, N111 , M113, V153, and N155 relative to the WT IL- 18 amino acid sequence set forth in SEQ ID NO:1 , including e.g., at least 1 , at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least, 8, at least 9, at least 10, at least 11 , or at least 12 of the listed mutated positions. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0252] Accordingly, useful DR-18 polypeptides can have one or any combination of mutations at the listed positions, with or without additional mutated residues at other positions. For example, useful polypeptides can have a mutation at position Y1, L5, K8, C38, M51 , K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, D110, N111, M113, V153, or N155 only; mutations at all positions Y1 , L5, K8, C38, M51 , K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, D110, N111 , M113, V153, and N155; or any combinations having more than one but less than all (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18 or 19) mutations at these positions.

[0253] In some instances, the DR-18 polypeptide comprises at least 12 mutations, at least 11 mutations, at least 10 mutations, at least 9 mutations, at least 8 mutations, at least 7 mutations, at least 6 mutations, at least 5 mutations, at least 4 mutations, at least 3 mutations, at least 2 mutations, or at least 1 mutation relative to the wild-type IL-18 amino acid sequence set forth in SEQ ID NO:1 , optionally wherein the at least 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 mutations are at positions selected from Y1 , L5, K8, C38, M51 , K53, S55, Q56, P57, G59, M60, C68, E77, Q103, S105, D110, N111 , M113, V153, and N155.

[0254] In some instances, the DR-18 polypeptide has less than 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, or 85% sequence identity to WT human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXXAVTISV KXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEGYFLAXEKERD LFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO: 64), wherein the X at pos. 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H.

[0255] In some instances, the DR-18 polypeptide comprises one or more substitutions relative to WT human IL-18 (SEQ ID NO: 1), including 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10 or more substitutions selected from: Y1 H, Y1 R, L5H, L5I, L5Y, K8Q, K8R, M51T, M51 K, M51 D, M51 N, M51 E, M51 R, K53R, K53G, K53S, K53T, S55K, S55R, Q56E, Q56A, Q56R, Q56V, Q56G, Q56K, Q56L, P57L, P57G, P57A, P57K, G59T, G59A, M60K, M60Q, M60R, M60L, E77D, Q103E, Q103K, Q103P, Q103A, Q103R, S105R, S105D, S105K, S105N, S105A, D110H, D110K, D110N, Attorney Docket No.: S199207 1060WO (00093)

[0256] ST-020-W01

[0257] D110Q, D110E, D110S, D110G, N111H, N111Y, N111 D, N111R, N111S, N111G, M113V, M113R, M113T, M113K, V153I, V153T, V153A, N155K, and N155H.

[0258] In some instances, the DR-18 polypeptide comprises one or more substitutions relative to WT human IL-18 (SEQ ID NO: 1) such that, e.g., the amino acid at pos. 1 is not Y and, e.g., may be R or H; the amino acid at pos. 5 is not L and, e.g., may be H, I or Y; the amino acid at pos. 8 is not K and, e.g., may be Q or R; the amino acid at pos. 38 is not C and, e.g., may be S; the amino acid at pos. 51 is not M and, e.g., may be T, K, D, N, E or R; the amino acid at pos. 53 is not K and, e.g., may be R, G, S or T; the amino acid at pos. 55 is not S and, e.g., may be K or R; the amino acid at pos. 56 is not Q and, e.g., may be E, A, R, V, G, K, L or R; the amino acid at pos. 57 is not P and, e.g., may be L, G, A or K; the amino acid at pos. 59 is not G and, e.g., may be A or T ; the amino acid at pos. 60 is not M and, e.g., may be K, Q, R or L; the amino acid at pos. 68 is not C and, e.g., may be S, G, A, V, D, E or N ; the amino acid at pos. 76 is not C and, e.g., may be S; the amino acid at pos. 77 is not E and, e.g., may be D; the amino acid at pos. 103 is not Q and, e.g., may be E, K, P, A or R; the amino acid at pos. 105 is not S and, e.g., may be D, N, R, K or A; the amino acid at pos. 110 is not D and, e.g., may be K, H, N, Q, E, S or G; the amino acid at pos. 111 is not N and, e.g., may be H, Y, D, R, S or G; the amino acid at pos. 113 is not M and, e.g., may be V, R, T or K; the amino acid at pos. 127 is not C and, e.g., may be S; the amino acid at pos. 153 is not V and, e.g., may be I, T or A; and / or the amino acid at pos. 155 is not N and, e.g., may be K or H.

[0259] In some instances, the DR-18 polypeptide comprises substitution mutations, relative to WT human IL-18 as set forth in SEQ ID NO:1 , at positions: (i) M51 , M60, S105, D110, and N111 ; (ii) M51 , S55, G59, M60, S105, D110, N111, and V153; (iii) Y1, M51 , M60, S105, D110, and N111 ; (iv) Y1 , M51 , K53, M60, S105, D110, and N111 ; (v) K8, M51 , S55, G59, M60, S105, D110, and N155; (vi) K8, M51 , S55, G59, M60, S105, D110, N111 , and V153; (vii) L5, M51 , K53, M60, S105, D110, and V153; (viii) L5, M51 , S55, G59, M60, S105, D110, N111 , and N155; (ix) L5, M51 , S55, M60, Q103, S105, D110, N111 , and V153; (x) L5, M51 , S55, M60, S105, D110, N111 , V153, and N155; (xi) L5, M51 , S55, G59, M60, S105, D110, N111 , V153, and N155; (xii) L5, K8, M51 , S55, M60, S105, N111 , V153, and N155; (xiii) L5, K8, M51 , K53, M60, S105, D110, N111 , and N155; (xiv) Y1 , L5, M51 , K53, M60, S105, D110, and N155; (xv) Y1 , M51, K53, G59, M60, S105, D110, N111 , V153, and N155; (xvi) Y1 , K8, M51 , K53, M60, Q103, S105, D110, N111, and N155; (xvii) Y1 , K8, M51 , M60, S105, D110, and N111 ; (xviii) Y1, L5, M51, K53, M60, Q103, S105, D110, and N111 ; (xix) Y1 , K8, M51 , K53, G59, M60, Q103, S105, D110, N111 , V153, and N155; (xx) Y1 , K8, M51 , K53, G59, M60, S105, D110, N111 , and N155; (xxi) Y1, K8, M51 , G59, M60, Q103, S105, D110, N111 , V153, and N155; (xxii) Y1 , L5, M51 , Attorney Docket No.: S199207 1060WO (00093)

[0260] ST-020-W01

[0261] G59, M60, E77, S105, D110, and N111 ; (xxiii) M51, Q56, P57, M60, Q103, S105, D110, N111 , and M113; (xxiv) M51 , Q56, P57, M60, Q103, S105, D110, and M113; (xxv) M51, K53, Q56, P57, M60, D110, and N111; (xxvi) M51 , K53, Q56, P57, M60, Q103, S105, D110, N111 , and M113; (xxvii) M51 , K53, Q56, M60, Q103, S105, D110, N111 , and M113; (xxviii) M51 , K53, Q56, P57, Q103, S105, D110, N111 , and M113; (xxix) M51 , K53, Q56, P57, M60, S105, D110, and N111 ; (xxx) M51 , K53, Q56, P57, M60, Q103, D110, N111 , and M113; (xxxi) M51 , Q56, P57, M60, Q103, D110, N111 , and M113; or (xxxii) M51 , K53, Q56, S105, D110, and N111.

[0262] In some instances, the DR-18 polypeptide comprises substitution, relative to WT human IL-18 as set forth in SEQ ID NO:1 , of: (i) M51T, M60K, S105D, D110K, and N111 H; (ii) M51T, S55K, G59A, M60K, S105D, D110K, N111 H, and V153I; (iii) Y1 R, M51T, M60K, S105D, D110K, and N111H; (iv) Y1 R, M51T, K53R, M60K, S105N, D110K, and N111Y; (v) K8Q, M51T, S55K, G59T, M60K, S105R, D110H, and N155K; (vi) K8R, M51 K, S55K, G59A, M60Q, S105D, D110K, N111 H, and V153I; (vii) K8R, M51 D, S55K, G59A, M60X, S105D, D110K, N111 H, and V153I; (viii) L5H, M51T, K53R, M60K, S105D, D110N, and V153T; (ix) L5I, M51K, S55K, G59A, M60Q, S105K, D110Q, N111 H, and N155K; (x) L5I, M51T, S55R, M60K, Q103E, S105D, D110H, N111 H, and V153I; (xi) L5I, M51T, S55K, M60K, S105D, D110K, N111 H, V153T, and N155H; (xii) L5I, M51T, S55K, G59A, M60K, S105R, D110H, N111H, V153I, and N155K; (xiii) L5I, K8R, M51T, S55K, M60K, S105D, N111Y, V153I, and N155K; (xiv) L5Y, K8R, M51T, K53R, M60K, S105D, D110E, N111 H, and N155K; (xv) Y1 H, L5Y, M51T, K53R, M60K, S105D, D110H, and N155K; (xvi) Y1 R, M51T, K53R, G59A, M60K, S105D, D110Q, N111H, V153A, and N155K; (xvii) Y1 R, K8R, M51 D, K53R, M60R, Q103K, S105N, D110K, N111Y, and N155H; (xviii) Y1 R, K8R, M51 N, K53R, M60Q, Q103K, S105R, D110N, N111H, and N155K; (xix) Y1 R, K8R, M51T, M60K, S105D, D110K, and N111 H; (xx) Y1R, L5H, M51T, K53R, M60K, Q103E, S105N, D110K, and N111Y; (xxi) Y1R, K8R, M51T, K53R, G59A, M60K, Q103E, S105D, D110Q, N111 H, V153I, and N155X; (xxii) Y1 R, K8R, M51T, K53R, G59T, M60K, S105N, D110H, N111 D, and N155H; (xxiii) Y1 R, K8R, M51T, G59A, M60K, Q103E, S105D, D110Q, N111 H, V153I, and N155K; (xxiv) Y1 R, L5Y, M51T, G59T, M60K, E77D, S105D, D110K, and N111H; (xxv) Y1R, K8R, M51T, K53R, G59T, M60K, S105K, D110N, N111 H, and N155K; (xxvi) M51E, Q56E, P57L, M60R, Q103P, S105A, D110N, N111R, and M113V; (xxvii) M51 K, Q56A, P57G, M60L, Q103E, S105D, D110S, and M113V; (xxviii) M51 K, K53G, Q56A, P57A, M60L, D110K, and N111R; (xxix) M51K, K53G, Q56R, P57G, M60L, Q103E, S105D, D110N, N111S, and M113R; (xxx) M51 K, K53G, Q56V, M60L, Q103A, S105A, D110S, N111 R, and M113T; (xxxi) M51K, K53S, Q56G, P57A, M60L, Q103A, S105A, D110G, N111 R, and M113T; (xxxii) M51K, K53S, Q56K, P57A, Q103A, S105D, D110S, N111S, and M113R; (xxxiii) M51K, K53S, Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0263] Q56L, P57A, M60L, S105D, D110S, and N111R; (xxxiv) M51 K, K53S, Q56R, P57A, M60L, S105N, D110G, and N111R; (xxxv) M51 K, K53S, Q56R, P57A, M60L, Q103A, D110G, N111 R, and M113T; (xxxvi) M51 K, K53S, Q56R, P57A, M60L, Q103A, S105D, D110S, N111G, and M113R; (xxxvii) M51 K, K53T, Q56R, M60L, Q103E, S105D, D110S, N111S, and M113K; (xxxviii) M51 K, K53T, Q56R, P57A, Q103E, S105D, D110N, N111 D, and M113R; (xxxix) M51 R, Q56G, P57K, M60L, Q103R, D110S, N111 R, and M113V; (xl) M51 K, K53G, Q56G, P57A, M60L, Q103E, S105D, D110S, N111G, and M113V; or (xli) M51 K, K53G, Q56R, S105A, D110N, and N111 R.

[0264] In some embodiments, the DR IL-18 polypeptide comprises: (i) an amino acid sequence having 85% or more (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% or 98%) sequence identity with the any one of the amino sequences set forth in SEQ ID NO: 2-63; and (ii) one or more mutations at a wild-type cysteine present in WT IL-18 as set forth in SEQ ID NO: 1 , such as e.g., C38, C68, C76, and / or C127. For example, in some instances, the DR IL-18 polypeptide comprises substitutions at amino acid positions Cysteine-38 and Cysteine-68 relative to WT human IL-18 SEQ ID NO: 1. In some instances, DR-18 polypeptide includes a stabilized DR-18 polypeptide. A stabilized DR-18 polypeptide can include mutations of two cysteine residues (e.g., C38 and C68) relative to human WT IL-18 (SEQ ID NO: 1). In some cases, the mutation is a C to S substitution and as such the stabilized IL-18 polypeptide can in some cases comprise the mutations C38S and C68S. In some cases a stabilized IL-18 polypeptide comprises the mutation pair C38S / C68S, C38S / C68G, C38S / C68A, C38S / C68V, C38S / C68D, C38S / C68E, or C38S / C68N (e.g., in some cases C38S / C68G, C38S / C68A, C38S / C68V, C38S / C68D, C38S / C68E, or C38S / C68N; in some cases C38S / C68S, C38S / C68G, C38S / C68A, C38S / C68D, or C38S / C68N; and in some cases C38S / C68G, C38S / C68A, C38S / C68D, or C38S / C68N).

[0265] DR-18 polypeptides can, additionally, be stabilized variants, through the presence of one or more stabilizing mutations. Useful DR-18 polypeptides, including in stabilized and nonstabilized forms, include those described in PCT Pub. Nos. WQ2019 / 051015 and WQ2022 / 094473, the disclosures of which are incorporated herein by reference in their entirety.

[0266] Useful IL-18 variants, as well as useful mutations and substitutions thereof, include but are not limited to those variants, mutations and substitutions described in WQ2002101049 (such as e.g., variants having mutations at positions E42 and / or K89 and substitutions E42A and K89A (numbering according to SEQ ID NO:65)), US20080206189 (such as e.g., variants having mutations at one or more of C38, C68, C76, N78, E121 , C127, L144, and D157 and substitutions, individually or in combination, of C38S, C68D, C68S, N78C, E121C, L144C, and Attorney Docket No.: S199207 1060WO (00093)

[0267] ST-020-W01

[0268] D157C), WO2023118497 (such as e.g., variants having mutations at one or more of Y1 , G3, S10, C38, M51 , K53, S55, P57, M60, C68, Q103, M113, C127, and N155, and substitutions, individually or in combination, of Y1A, G3Y, S10R, S10K, C38S, M51A, M51Y, M51Q, K53L, K53H, K53A, S55L, S55Y, P57T, M60A, M60E, M60H, C68S, Q103S, M113A, M113E, C127S, and N155Y (inc. those at pos. K53 other than K53R, K53G, K53S, or K53T), WO2023114829 (such as e.g., variants having mutations at one or more of Y37, S43, L45, S46, V47, N50, F57, C74, T81, I85, S86, D90, S91 , A97, V98, T99, S101 , C104, 1107, T109, C112, N123, P124, Q139, R140, Q150, A162, C163, L174, 1185, V189, Q190, and E192, and substitutions, individually or in combination, of Y37C, S43C, L45C, S46C, V47C, N50C, F57C, C74S, T81C, I85C, S86C, D90C, S91C, A97C, V98C, T99C, S101C, C104S, I107C, T109C, C112S, N123C, P124C, Q139C, R140C, Q150C, A162C, C163S, L174C, I185C, V189C, Q190C, E192C, (numbering according to SEQ ID NO:65)), WO2023056193 (such as e.g., variants having mutations at one or more of M51, K53, Q56, P57 and M60, and substitutions, individually or in combination, of M51A, M51S, M51G, K53G, K53A, K53S, Q56R, Q56K, Q56H, P57A, P57S, P57G, M60R, M60K, and M60H (and useful sequences encoding such variants, such as e.g., SEQ ID NOs: 7 and 8 of WO2023056193)), US20230146665 (such as e.g., variants having mutations at one or more of Y1, E6, S7, K8, S10, V11 , N14, L15, D17, Q18, D23, R27, P28, L29, E31 , M33, T34, D35, S36, D37, C38, R39, D40, N41 , R44, I46, I49, S50, M51 , K53, D54, S55, Q56, P57, M60, A61, V62, T63, S65, K67, C68, E69, 171 , C76, E77, I80, 181 , N87, P88, D90, K93, T95, K96, S97, Q103, H109, D110, N111 , M113, S119, A126, C127, D132, L136, L138, K139, E141 , L144, D146, R147, 1149, M150, N155, E156, and D157, and substitutions, individually or in combination, of Y1 F, Y1 H, E6A, E6Q, S7C, S7P, K8E, K8Q, K8Y, S10C, V11I, N14C, N14W, L15C, D17N, Q18L, D23N, D23S, R27Q, P28C, L29V, E31Q, M33C, T34P, D35N, D35E, S36D, S36N, D37N, C38S, C38Q, C38R, C38E, C38L, C38I, C38V, C38K, C38D, R39S, R39T, D40N, N41Q, R44Q, I46V, I49C, S50C, S50Y, M51I, M51 K, M51Q, M51R, M51L, M51H, M51 F, M51Y, K53A, K53D, K53E, K53G, K53H, K53I, K53L, K53M, K53N, K53Q, K53R, K53S, K53T, K53V, K53Y, K53F, D54C, S55N, S55Q, S55D, S55E, S55T, Q56I, Q56L, P57A, P57E, P57T, P57V, P57Q, P57D, P57Y, P57N, M60I, M60L, M60K, M60Y, M60F, A61C, V62C, T63C, S65C, K67Q, C68S, C68I, C68F, C68Y, C68D, C68N, C68E, C68Q, C68K, E69K, 171 M, C76S, C76E, C76K, E77K, 180T, 181 L, 181V, N87S, P88C, D90E, K93D, K93N, T95E, K96G, K96Q, S97N, Q103C, Q103E, Q103I, Q103L, H109W, H109Y, D110N, D110Q, D110R, N111 D, N111Q, N111S, N111T, N111 E, M113I, S119L, A126C, C127S, C127W, C127Y, C127F, C127D, C127E, C127K, D132Q, D132E, L136C, L138C, K139C, E141K, E141Q, L144N, D146F, D146L, D146Y, R147C, R147K, 1149V, M150F, M150T, N155C, E156Q, D157A, Attorney Docket No.: S199207 1060WO (00093)

[0269] ST-020-W01

[0270] D157S, and D157N, as well as deletion mutants described therein), US20220056091 (such as e.g., variants having mutations at one or more of Y01 , F02, E06, M33, C38, M51 , K53, D54, S55, M60, T63, C68, E69, K70, C76, M86, M113, C127, and M150, and substitutions, individually or in combination, of Y01G, F02A, E06K, C38A, C38S, K53A, D54A, S55A, T63A, C68A, C68S, E69C, K70C, C76A, C76S, C127A, and C127S), US20230357342 (such as e.g., variants having mutations at one or more of Y1 , F2, E6, K8, S10, V11 , D17, E31 , T34, D35, S36, D37, C38, D40, N41 , I49, M51 , K53, D54, S55, T63, C68, E69, K70, C76, Q103, S105, G108, H109, D110, C127, D132, and V153, and substitutions, individually or in combination, of Y1M, Y1G, F2A, E6K, E6R, K8E, K8L, K8R, S10T, V111, D17N, E31A, T34A, D35A, S36A, D37A, C38A, C38Q, C38S, D40A, N41A, I49E, I49M, I49R, M51G, K53A, D54A, S55A, S55H, S55R, S55T, T63A, C68A, C68S, E69C, K70C, C76A, C76S, Q103E, Q103K, Q103R, S105I, S105K, G108A, H109A, D110A, C127A, C127S, D132A, V153E, V153R, and V153Y), and WO2022172944 (such as e.g., variants having mutations at one or more of C38, C68, C76, C127, G3, A6, S7, T34, S50, M51, S72, K112, K119, and G145, and substitutions, individually or in combination, of C38S, C68S, C76S, C127S, G3Y, G3L, A6W, S7C, T34P, C38M, S50C, M51Y, S72Y, S72F, S72M, S72L, S72W, K112W, K119V, and G145N).

[0271] A nucleic acid sequence encoding a DR-18 polypeptide may be generated from the DR- 18 polypeptide amino acid sequence, including e.g., any of the DR-18 polypeptide amino acid sequences described herein. Such DR-18 polypeptide-encoding sequences may be produced by conversion of each amino acid residue to a corresponding trinucleotide codon that codes for the residue. Such DR-18 polypeptide-encoding sequences may include a stop codon after the codon designating the terminal amino acid. In some instances, a stop codon may be added.

[0272] In some instances, DR-18 polypeptide encoding sequences of the present disclosure may be codon optimized, i.e., optimized for expression in a particular host cell or organism. In some instances, a DR-18 polypeptide encoding sequence of the present disclosure is optimized for expression in human cells, i.e., is human codon optimized. Useful methods for codon optimization, including human codon optimization, include but are not limited to e.g., those described in US20140244228, US20210366574, US20190325989, US20230245721 , US20160259885, US20140377800, W02020024917, WO2022221576, W02024018050, and W02009009743.

[0273] As summarized above, useful agents in the herein described methods include monospecific antibodies, multi-specific therapeutics, such as bispecific, trispecific, and tetraspecific antibodies, and the like. Such agents target and specifically bind to one or more antigens. Such antigens may include antigens expressed on cancer cells, antigens expressed Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 on non-cancerous cells, antigens expressed on healthy cells, antigens expressed on immune cells, such as immune effector cells, combinations thereof, and the like.

[0274] In some embodiments, useful antigens (e.g., antigens to which an antibody will specifically bind) include antigens associated with B cell cancer. Useful antigens associated with B cell cancer include, but are not necessarily limited to, e.g., CD19, CD20, CD22, CD23, CD79a, and CD79b.

[0275] In some embodiments, useful antigens (e.g., antigens to which an antibody will specifically bind) include antigens associated with plasma cell cancer. In some embodiments, useful antigens (e.g., antigens to which an antibody will specifically bind) include antigens associated with multiple myeloma. Useful antigens associated with plasma cell cancer and / or multiple myeloma, include but are not necessarily limited to, e.g., CD38, CD138, CD56, BCMA (B-cell maturation antigen), SLAMF7 (Signaling Lymphocytic Activation Molecule F7), and CS1 (CD319).

[0276] In some embodiments, useful antigens, that may be associated with B cell and / or plasma cell cancers, include, e.g., CD10, CD30, CD45, CD52, and MUC1 (Mucin 1). In some embodiments, antibodies, including monoclonal, monospecific, multi-specific antibodies, and the like, target antigens associated with B cell cancer such as, e.g., CD19, CD20, CD22, CD23, CD79a, and CD79b.

[0277] In some embodiments, antibodies, including monospecific, multi-specific, bispecific antibodies, and the like, may be used to target antigens associated with plasma cell cancer. In some embodiments, antibodies, including monospecific, multi-specific, bispecific antibodies, and the like, may be used to target antigens associated with multiple myeloma. Such useful antibodies that may target plasma cell cancer and / or multiple myeloma include antibodies that specifically bind antigens such as, e.g., CD38, CD138, CD56, BCMA (B-cell maturation antigen), SLAMF7 (Signaling Lymphocytic Activation Molecule F7), and CS1 (CD319). Useful antigens for targeting multiple myeloma include, e.g., BCMA, CD38, FcRH5, CD19, GPRC5D, and CD138. Useful antibodies that target multiple myeloma include antibodies binding antigens such as, e.g., BCMA, CD38, FcRH5, CD19, GPRC5D, and CD138.

[0278] In some embodiments, antibodies, including monospecific, multi-specific, bispecific antibodies, and the like, may target antigens that are associated with B cell and / or plasma cell cancers. Such useful antibodies include, e.g., those antibodies that specifically bind to, e.g., CD10, CD30, CD45, CD52, and MUC1.

[0279] Useful antibodies, antigens, and antigen binding domains for targeting hematological cancers include e.g., CD19 as bound by the antigen binding domains of Tisagenlecleucel and Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0280] Axicabtagene ciloleucel and used in the treatment of B-cell acute lymphoblastic leukemia (IB- ALL) and DLBCL; CD20 as bound by the antigen binding domains of Rituximab, Obinutuzumab, Ofatumumab and used in the treatment of Non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL); CD22 as bound by the antigen binding domain of Inotuzumab ozogamicin and used to treat B-ALL; CD33 as bound by the antigen binding domain of Gemtuzumab ozogamicin and used to treat AML; CD52 as bound by the antigen binding domain of Alemtuzumab and used to treat AML; CD52 CLL and T-cell prolymphocytic leukemia (T-PLL); CD38 as bound by the antigen binding domains of Daratumumab and Isatuximab and used to treat multiple myeloma; CD30 as bound by the antigen binding domain of Brentuximab vedotin and used to treat Hodgkin lymphoma and Systemic anaplastic large cell lymphoma (sALCL); CD47 as bound by the antigen binding domain of Magrolimab and used to treat AML and various solid tumors; CD123 as bound by the antigen binding domain of Talacotuzumab and used to treat AML and blastic plasmacytoid dendritic cell neoplasm (BPDCN); and SLAMF7 (CS1) as bound by the antigen binding domain of Elotuzumab and used to treat Multiple myeloma. As will be readily understood, such antigens may be targeted in a multi-specific therapeutic, where e.g., such a multi-specific therapeutic employs the antigen binding domain of a monospecific antibody or chimeric antigen receptor, such as those described here, in a bispecific architecture with a second antigen binding domain binding a second antigen, such as e.g., an immune cell antigen, such as e.g., CD3.

[0281] In some embodiments, antibodies, including monospecific, multi-specific, bispecific antibodies, and the like, may target antigens that are associated with solid cancers. Useful antigens that can be targeted in solid tumors include but are not limited to e.g., HER2 / neu (ERBB2) as targeted by trastuzumab and pertuzumab for the treatment of breast cancer and gastric cancer; EGFR (Epidermal Growth Factor Receptor) as targeted by cetuximab and panitumumab in the treatment of colorectal cancer and head and neck cancer; PD-L1 (Programmed Death-Ligand 1) as targeted by atezolizumab, durvalumab, and avelumab in the treatment of lung cancer, bladder cancer, and Merkel cell carcinoma; CD20 as targeted by obinutuzumab and rituximab in the treatment of glioma in clinical trials; GD2 as targeted by dinutuximab in the treatment of neuroblastoma; CTLA-4 as targeted by ipilimumab in the treatment of melanoma and renal cell carcinoma; CA-125 (also known as MUC16) as targeted by oregovomab in the treatment of ovarian cancer; Glypican-3 (GPC3) as targeted by codrituzumab in the treatment of hepatocellular carcinoma; CLDN18.2 as targeted by zolbetuximab in the treatment of gastric cancer, gastroesophageal junction cancer, and pancreatic ductal adenocarcinoma; and TRP-1 (Tyrosinase-related protein 1, a.k.a. TYRP-1 and Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 gp75) as targeted by flanvotumab (IMC-20D7S) in the treatment of melanoma in clinical trials. As will be readily understood, such antigens may be targeted in a multi-specific therapeutic, where e.g., such a multi-specific therapeutic employs the antigen binding domain of a monospecific antibody, such as those described here, in a bispecific architecture with a second antigen binding domain binding a second antigen, such as e.g., an immune cell antigen, such as e.g., CD3.

[0282] In some embodiments, a multi-specific therapeutic will also include (i.e., in addition to a binding domain that binds a cancer antigen) an antigen binding domain that specifically binds to an antigen expressed on non-cancerous immune cells, which may include e.g., healthy immune cells, functional immune cells, and, in some instances, inactive immune cells, and including populations of cells that include combinations thereof, in the subject. In some instances, such immune cells within the subject are referred to as cells of the anti-cancer immune cell population. In some instances, such immune cells within the subject are referred to as immune effector cells or effector immune cells, such as e.g., effector T cells. In some instances, anticancer immune cells may include the endogenous immune cells, e.g., endogenous T cells and / or endogenous NK cells, present within the subject’s immune system. In some instances, anti-cancer immune cells may include immune cells provided from an exogenous source, e.g., exogenous T cells and / or exogenous NK cells, administered to the subject, including e.g., through an adoptive cell therapy, including autologous or allogeneic adoptive cell therapies. In some instances, anti-cancer immune cells may be derived from transplanted bone marrow or transplanted hematopoietic stem cells, including where such transplants are autologous or allogeneic.

[0283] In some instances, immune cells within the subject are inactive (or under-active) and combination therapy through administration of a DR-18 and a multi-specific therapeutic that targets at least one antigen expressed on the inactive immune cells is capable of activating (or enhancing the activation of) the cells to treat the subject for cancer. Immune cell populations that can be targeted include e.g., T cells, NK cells, and the like. Inactive (or under-active) immune cells may display one or more characteristics of immune cell exhaustion. Immune cell exhaustion is characterized by progressive loss of effector function; expression of inhibitory receptors such as PD-1 , CTLA-4, LAG-3, and TIM-3 on T cells and KLRG1 and NKG2A on NK cells; altered transcriptional profiles; diminished cytokine production; impaired proliferative capacity; metabolic dysregulation, including altered mitochondrial function and a reliance on fatty acid oxidation over glycolysis; and the like. Active, or re-activated, immune cells generally display characteristics opposite those of inactive or exhausted immune cells, such as e.g., Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 active effector function; reduced expression of inhibitory receptors such as PD-1 , CTLA-4, LAG- 3, and TIM-3 on T cells and KLRG1 and NKG2A on NK cells; normal / healthy transcriptional profiles; normal or increased cytokine production; normal or increased proliferative capacity; normal metabolic activity; and the like. In some instances, activated (or re-activated) immune cells may be characterized by expression of cytokines, such as IL-2, IFN-y, and TNF-a; reduced expression of PD-1 and / or CTLA-4; expression of co-stimulatory molecules, such as e.g., CD28 and CD137 (4-1 BB); expression of differentiation and persistence markers such as e.g., T-bet, Eomesodermin, and the like; active glycolysis; reduced fatty acid oxidation; and the like. Expression of markers indicative of inactive or active immune cells may be measured by any convenient means, including but not limited to e.g., flow cytometry, PCR, NGS, and the like.

[0284] Useful antigens expressed on immune cells that can be targeted in multi-specific therapeutics, such as bispecific and trispecific antibodies, include but are not limited to CD3, CD16, CD28, and the like. CD3 is a complex of proteins found on T cells and plays a crucial role in activating both cytotoxic and helper T cells. CD3-targeting therapeutics can engage T cells against cancer cells by facilitating the formation of immune synapses and promoting anti-tumor cytotoxic activity. CD16, also known as Fc gamma RIH, is found mainly on Natural Killer (NK) cells and certain subtypes of T cells. Targeting CD16 can enhance antibody-dependent cellular cytotoxicity (ADCC), leveraging NK cells for the killing of targeted cancer cells. CD28 is expressed on T cells and serves as a co-stimulatory molecule for T cell activation and survival. Targeting CD28 can facilitate the activation and expansion of T cells upon antigen recognition, thereby enhancing the body’s immune response against cancer cells.

[0285] Antigens expressed on functional immune cells present in a subject can be employed as targets in therapeutic antibodies. Multi-specific therapeutics, such as e.g., bispecific and trispecific antibodies, can target antigens expressed on healthy immune cells to, e.g., bridge the interaction between immune cells and cancer cells, leading to enhanced immune-mediated destruction of malignancies.

[0286] In some embodiments, useful therapeutic monospecific antibodies targeting hematological cancers include Rituximab, Obinutuzumab, Ofatumumab, Alemtuzumab, Daratumumab, Elotuzumab, Gemtuzumab ozogamicin, Ibritumomab tiuxetan, Tositumomab, and Brentuximab vedotin. Monospecific antibodies like Rituximab, Obinutuzumab, and Ofatumumab may specifically target CD20 on B-cells, while Alemtuzumab targets CD52, and Daratumumab and Elotuzumab target SLAMF7 on plasma cells. Attorney Docket No.: S199207 1060WO (00093)

[0287] ST-020-W01

[0288] In some embodiments, such monospecific antibodies may be utilized in the treatment of various hematological malignancies such as Hodgkin lymphoma, non-Hodgkin lymphoma, chronic lymphocytic leukemia, and multiple myeloma, among others.

[0289] In some embodiments, the therapeutic application of monospecific antibodies may involve mechanisms such as complement-dependent cytotoxicity, antibody-dependent cell- mediated cytotoxicity, or direct induction of apoptosis in malignant cells.

[0290] In some embodiments, multi-specific therapeutic antibodies, including bispecific and trispecific antibodies, can be utilized in the methods of the disclosure. These multi-specific antibodies are designed to simultaneously bind to two or more distinct antigens. For example, by targeting both of at least a first and a second antigen, the multi-specific therapeutic may facilitate the recruitment and activation of immune cells to target cancer cells. For instance, bispecific antibodies may possess one antigen-binding region specific for a tumor-associated antigen and another for an antigen expressed on immune cells, such as T cells or NK cells, effectively bridging the two cell types and promoting targeted cytotoxicity. Trispecific antibodies, featuring three antigen-binding regions, may further enhance the specificity and / or efficacy of the immune response. Such multi-specific antibodies can be employed to enhance the therapeutic efficacy of DR-18 by synergizing with its immune-modulating effects, thereby improving clinical outcomes in the treatment of cancers, such as hematological malignancies.

[0291] In some embodiments, a multi-specific therapeutic antibody targeting at least CD19 and CD3 may be used, such as e.g., in the treatment of hematological cancers. In some embodiments, a bispecific therapeutic antibody targeting CD19 and CD3 may be used in the treatment of hematological cancers. For example, blinatumomab is an example of such an antibody, used in treating B-cell precursor acute lymphoblastic leukemia (ALL). Blinatumomab has been investigated in clinical trials such as NCT02013167, which demonstrated its efficacy in pediatric patients with ALL (Kantarjian et al., (2017) N Engl J Med 376(9): 836-847).

[0292] In some embodiments, a multi-specific antibody targeting at least CD22 and CD3 may be employed, such as e.g., for the treatment of hematological cancers. In some embodiments, a bispecific antibody targeting CD22 and CD3 may be employed for the treatment of hematological cancers. An example of a therapeutic targeting CD22 is moxetumomab pasudotox, which has been used for treating hairy cell leukemia. Clinical trials, including NCT01829711. The CD22-binding portion of moxetumomab pasudotox is useful in the development of CD22 bispecific antibodies, such as a CD22 / CD3 bispecific.

[0293] In some embodiments, a multi-specific antibody targeting at least BCMA (B-cell maturation antigen) and CD3 may be utilized, such as e.g., in the treatment of multiple Attorney Docket No.: S199207 1060WO (00093)

[0294] ST-020-W01 myeloma. In some embodiments, a bispecific antibody targeting BCMA and CD3 may be utilized in the treatment of multiple myeloma. An example of such a therapeutic is teclistamab, and its effectiveness and safety have been investigated in clinical trials such as NCT04557098, which includes patients with relapsed or refractory multiple myeloma (Usmani et al., 2021 Lancet 398(10301):665-674).

[0295] In some embodiments, a multi-specific antibody targeting at least FLT3 and CD3 may be used, such as e.g., for the treatment of acute myeloid leukemia (AML). In some embodiments, a bispecific antibody targeting FLT3 and CD3 may be used for the treatment of AML. The compound AMG 427 is an example of such an antibody and is currently in clinical trials such as NCT03541369 to assess its safety and efficacy in patients with AML.

[0296] In some embodiments, a multi-specific antibody targeting at least CD123 and CD3 may be employed, e.g., for the treatment of hematological malignancies. In some embodiments, a bispecific antibody targeting CD123 and CD3 may be employed for the treatment of hematological malignancies. An example of this is flotetuzumab, which has been evaluated for effectiveness in the treatment of AML through clinical trials such as NCT02152956 (Frankel et al., 2020). Another example is a next generation version of flotetuzumab known as MGD024.

[0297] In some embodiments, a multi-specific antibody that targets at least CD20 and CD3 may be utilized, e.g., for the treatment of B-cell malignancies. In some embodiments, a bispecific antibody that targets CD20 and CD3 may be utilized for the treatment of B-cell malignancies. Mosunetuzumab is an example of this class of antibodies, evaluated in clinical trials like NCT02500407 for patients with relapsed or refractory B-cell non-Hodgkin lymphoma (Budde et al., 2021 Lancet Oncol 23(8): 1055-1065). Epcoritamab is another example, evaluated in clinical trials like NCT03625037 for patients with relapsed and / or refractory B-cell lymphoma. GB261 is another example, evaluated clinical trials like NCT04923048 in patients with relapsed / refractory non-Hodgkin lymphoma and chronic lymphocytic leukemia (see also Song et al., Blood (2023) 142(supplement 1): 1719). Odronextamab is another example, evaluated in clinical trials like NCT02290951 in patients with B-cell malignancies. Glofitamab is another example, evaluated clinical trials like NCT05896163 in patients with relapsed or refractory diffuse large B cell lymphoma.

[0298] In some embodiments, a multi-specific antibody targeting at least CD33 and CD3 may be utilized, e.g., in the treatment of acute myeloid leukemia. In some embodiments, a bispecific antibody targeting CD33 and CD3 may be applied in the treatment of acute myeloid leukemia. AMG 330 is an example currently being explored in clinical trials such as NCT02520427, which aim to assess its safety profile and therapeutic potential in AML patients. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0299] Useful multi-specific therapeutics for the treatment of multiple myeloma include e.g., BiTE's targeting BCMA, CD38, FcRH5, CD19, CD138, and the like, such as e.g., those described herein, those described in Alhallak et al., Cancers (2021) 13(12):2853, and the like. Useful multi-specific therapeutics for the treatment of multiple myeloma include e.g., BiTE's targeting GPRC5D, such as e.g., talquetamab as described in Chari et al. (2022) N Engl J Med 2022;387:2232-2244.

[0300] Useful multi-specific therapeutics useful in combination with DR-18 in the methods of the present disclosure include those that target CD3 and at least one other antigen, such as e.g., multi-specific antibodies targeting the following antigen combinations: CD3 x gp100, CD3 x DLL3, CD3 x PRAME, CD3 x STEAP1, CD3 x MUC16, CD3 x DLL3, CD3 x PIWIL1, CD3E X DLL3 x albumin, CD3 x HER2, CD3 x CLDN18.2, CD3 x CLDN6, CD3 x HLA x MAGEA4, CD3 x PSMA, CD3 x CEA, CD3 x EGFR, CD3 x PRAME, CD3 x EpCAM, CD3 x EGFR, CD3 x PSMA, CD3 x GPC3, CD3 x CDH3 x MSLN, CD3 x CDH17, CD3 x CLDN18.2, CD3 x NCR3LG1 , CD3 x 5T4, CD3 x CLDN6, CD3 x CLDN6, CD3 x EGFR, CD3 x KLK2, CD3 x ENPP3, CD3 x CALR, CD3 x CEACAM5, CD3 x ROR1 , CD3 x EGFRvlll, CD3 x CD3c x HER2, CD3 x 4-1BB x DLL3, CD3 x CLDN6, CD3 x ENPP3, CD3 x MSLN, CD3 x PSMA, CD3 x MSLN, CD3 x CD20, CD3 x GPRC5D, CD3 x BCMA, CD3 x CD19, CD3 x BCMA, CD3 x CD20, CD3 x CD20, CD3 x CD20, CD3 x BCMA, CD3 x CD19 x CD3D, CD3 x CD123, CD3 x FCRL5, CD3 x BCMA, CD3 x GPRC5D, CD3 x CD123, CD3 x FLT3, CD3 x CD19, CD3 x CD38, CD3 x BCMA x CD38, CD3 x CD22, CD3 x BCMA x GPRC5D, CD3 x CD20 x CD79B, CD3 x CD123, CD3 x PD-1 , CD3 x BCMA, CD3 x CD19 x CD2, CD3 x CD38, CD3 x BCMA, CD3 x EpCAM, CD3 x MUC16, CD3 x DLL3, CD3 x GPC3, CD3 x CD19 x CD20 x HER2, CD3 x HER2, CD3 x TAA, CD3 x CD33, CD3 x CLDN18.2, CD3 x CD276, CD3 x EpCAM, CD3 x BCMA x GPRC5D, CD3 x CD19 x CD20, CD3 x BCMA, CD3 x CD20, CD3 x CD20, CD3 x BCMA x GPRC5D, CD3 x GPRC5D, CD3 x CD19 x CD28, CD3 x BCMA x GPRC5D, and CD3 x CD276.

[0301] Non-limiting examples of multi-specific therapeutics useful in combination with DR-18 in the methods of the present disclosure that target CD3 and at least one other antigen, including Tebentafusp (further targeting gp100, developed by Immunocore Ltd.), Tarlatamab (further targeting DLL3, developed by Amgen, Inc.), Brenetafusp (further targeting PRAME, developed by Immunocore Ltd.), Xaluritamig (further targeting STEAP1 , developed by Amgen, Inc.), Ubamatamab (further targeting MUC16, developed by Regeneron Pharmaceuticals, Inc.), Obrixtamig (further targeting DLL3, developed by Boehringer Ingelheim GmbH), IMC-R117C (further targeting PIWIL1 , developed by Immunocore Ltd.), Gocatamig (further targeting DLL3 x albumin, developed by Harpoon Therapeutics, Inc.), VIR-5818 (further targeting HER2, Attorney Docket No.: S199207 1060WO (00093)

[0302] ST-020-W01 developed by Amunix Pharmaceuticals, Inc.), AZD5863 (further targeting CLDN18.2, developed by Harbour Biomed Therapeutics Ltd.), BNT-142 (further targeting CLDN6, developed by BioNTech SE), CDR-404 (further targeting HLA x MAGEA4, developed by CDR-Life AG), REGN4336 (further targeting PSMA, developed by Regeneron Pharmaceuticals, Inc.), Cibisatamab (further targeting CEA, developed by F. Hoffmann-La Roche Ltd.), TAK-186 (further targeting EGFR, developed in partnership with Takeda Pharmaceutical Co., Ltd.), IMA- 402 (further targeting PRAME, developed by Immatics Biotechnologies GmbH), BA-3182 (further targeting EpCAM, developed by BioAtla, Inc.), CX-904 (further targeting EGFR, developed by Amgen, Inc. | CytomX Therapeutics, Inc.), JANX007 (further targeting PSMA, developed by Janux Therapeutics, Inc.), ERY-974 (further targeting GPC3, developed by Chugai Pharmaceutical Co., Ltd.), AMG-305 (further targeting CDH3 x MSLN, developed by Amgen, Inc.), Cabotamig (further targeting CDH17, developed by Arbele Corp.), ASP-2138 (further targeting CLDN18.2, developed by Astellas Pharma, Inc. | Xencor, Inc.), BI-765049 (further targeting NCR3LG1, developed by Boehringer Ingelheim GmbH), CBA-1535 (further targeting 5T4, developed by Chiome Bioscience, Inc.), SAIL-66 (further targeting CLDN6, developed by Chugai Pharmaceutical Co., Ltd.), CTIM-76 (further targeting CLDN6, developed by Integral Molecular, Inc.), JANX008 (further targeting EGFR, developed by Janux Therapeutics, Inc.), JNJ-78278343 (further targeting KLK2, developed by Janssen Research & Development LLC | Zymeworks BC, Inc.), JNJ-87890387 (further targeting ENPP3, developed by Janssen Research & Development LLC), JNJ-88549968 (further targeting CALR, developed by Janssen Research & Development LLC), NILK-2301 (further targeting CEACAM5, developed by Novimmune SA), NM32-2668 (further targeting ROR1 , developed by Numab Therapeutics AG), RO-7428731 (further targeting EGFRvlll, developed by Roche Holding AG), Runimotamab (further targeting CD3c x HER2, developed by F. Hoffmann-La Roche Ltd.), RO-7616789 (further targeting 4-1 BB x DLL3, developed by F. Hoffmann-La Roche Ltd.), XmAb541 (further targeting CLDN6, developed by Xencor, Inc.), XmAb-819 (further targeting ENPP3, developed by Xencor, Inc.), ZW-171 (further targeting MSLN, developed by Zymeworks BC, Inc.), VIR- 5500 (further targeting PSMA, developed by Vir Biotechnology, Inc.), CT-95 (further targeting MSLN, developed by Link Immunotherapeutics, Inc.), Epcoritamab (further targeting CD20, developed by Genmab A / S), Talquetamab (further targeting GPRC5D, developed by Janssen Pharmaceuticals, Inc.), Teclistamab (further targeting BCMA, developed by Janssen Research & Development LLC), Blinatumomab (further targeting CD19, developed by Amgen, Inc.), Elranatamab (further targeting BCMA, developed by Pfizer Inc.), Odronextamab (further targeting CD20, developed by Regeneron Pharmaceuticals, Inc.), Glofitamab (further targeting Attorney Docket No.: S199207 1060WO (00093)

[0303] ST-020-W01

[0304] CD20, developed by F. Hoffmann-La Roche Ltd.), Mosunetuzumab (further targeting CD20, developed by Genentech, Inc.), Etentamig (further targeting BCMA, developed by TeneoBio, Inc.), Surovatamig (further targeting CD19 x CD3D, developed by TeneoBio, Inc.), Vibecotamab (further targeting CD123, developed by Xencor, Inc.), Cevostamab (further targeting FCRL5, developed by Genentech, Inc.), REGN-5459 (further targeting BCMA, developed by Regeneron Pharmaceuticals, Inc.), Forimtamig (further targeting GPRC5D, developed in partnership with Roche - Genentech), Mipletamig (further targeting CD123, developed by Aptevo Therapeutics, Inc.), CLN-049 (further targeting FLT3, developed by German Cancer Research Center | Universitaetsklinikum Tuebingen), CLN-978 (further targeting CD19, developed by Adimab LLC), ISB-1342 (further targeting CD38, developed by Glenmark Pharmaceuticals Ltd.), ISB- 2001 (further targeting BCMA x CD38, developed by Ichnos Sciences, Inc.), Rezetamig (further targeting CD22, developed by TeneoBio, Inc.), JNJ-79635322 (further targeting BCMA x GPRC5D, developed by Janssen Research & Development LLC), JNJ-80948543 (further targeting CD20 x CD79B, developed by Janssen Research & Development LLC), MGD-024 (further targeting CD123, developed by MacroGenics, Inc.), ONO-4685 (further targeting PD-1, developed by Merus NV), WVT-078 (further targeting BCMA, developed by Novartis Pharma AG), PIT-565 (further targeting CD19 x CD2, developed by Novartis Pharmaceuticals Corp.), XmAb968 (further targeting CD38, developed by Xencor, Inc.), Linvoseltamab (further targeting BCMA, developed by Regeneron Pharmaceuticals, Inc.), M-701 (further targeting EpCAM, developed by Wuhan YZY Biopharma Co., Ltd.), LBL-033 (further targeting MUC16, developed by Nanjing Leads Biolabs Co., Ltd.), ZG-006 (further targeting DLL3, developed by Suzhou Zelgen Biopharmaceuticals Co., Ltd.), CM-350 (further targeting GPC3, developed by Keymed Biomedical Technology (Chengdu) Co., Ltd.), BR-115 (further targeting CD19 x CD20 x HER2, developed by Chengdu Enmu Biotechnology Co., Ltd), EX-101 (further targeting HER2, developed by Excelmab, Inc.), TGI-6 (further targeting TAA, developed by Hefei TG ImmunoPharma Co., Ltd), TTX-564 (further targeting CD33, developed by Trueline Therapeutics Inc.), IBI-389 (further targeting CLDN18.2, developed by Innovent Biologies (Suzhou) Co. Ltd.), EX-105 (further targeting CD276, developed by Excelmab, Inc.), Catumaxomab (further targeting EpCAM, developed by Neovii Biotech GmbH), MBS314 (further targeting BCMA x GPRC5D, developed by Beijing Mabworks Biotech Co., Ltd.), CMG1A46 (further targeting CD19 x CD20, developed by Chengdu Enmu Biotechnology Co., Ltd), Cizutamig (further targeting BCMA, developed by EpimAb Biotherapeutics, Inc.), EX-103 (further targeting CD20, developed by Excelmab, Inc.), GB-261 (further targeting CD20, developed by Genor Biopharma Co., Ltd.), IBI-3003 (further targeting BCMA x GPRC5D, Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 developed by Innovent Biologies (Suzhou) Co. Ltd.), LBL-034 (further targeting GPRC5D, developed by Nanjing Leads Biolabs Co., Ltd.), CC-312 (further targeting CD19 x CD28, developed by CytoCares Shanghai, Inc.), SIM-0500 (further targeting BCMA x GPRC5D, developed by Simcere Pharmaceutical Group Ltd.), and TAK-280 (further targeting CD276, developed by Maverick Therapeutics, Inc.).

[0305] In some embodiments, useful multi-specific antibodies may be trispecific. Useful trispecific antibodies include e.g., JNJ-80948543, a T-cell redirecting CD79b x CD20 x CD3 trispecific antibody as used in clinical trials such as NCT05424822 for the treatment of patients with non-Hodgkin lymphoma or chronic lymphocytic leukemia. Useful trispecific antibodies also include e.g., HPN217 (Harpoon Therapeutics) targeting T cells, MM, and albumin using anti- CD3, anti-BCMA, and anti-albumin, respectively.

[0306] In some embodiments, agents, such as antibodies, described herein can be employed in combination with a DR-18 or a nucleic acid encoding a DR-18, thereby providing enhanced therapeutic efficacy against cancers. Such, antibodies, whether monospecific, multi-specific, bispecific, or trispecific, are capable of targeting cancer-associated antigens and may work synergistically with DR-18 to augment the anti-cancer immune response.

[0307] Nucleic acids encoding DR-18 can be co-administered (with a relevant antibody) via appropriate delivery mechanisms, such as viral vectors, non-viral vectors (e.g., lipid nanoparticles), or directly as DNA (e.g., plasmid DNA) or mRNA. The expressed DR-18 polypeptide from these nucleic acids will activate the immune cells, thereby enhancing the activity of concurrently administered therapeutic antibodies.

[0308] Kits

[0309] The present invention also pertains to kits useful in the methods of the disclosure. Such kits comprise various combinations of components useful in any of the methods described elsewhere herein, including for example, a DR-18 polypeptide or encoding nucleic acid, materials for combination therapy, materials for performing analysis for patient selection, and / or materials for assessing the activity of one or more administered agents, and / or instructional material.

[0310] Any of the components, agents, materials, devices, carriers, and the like, described herein may be included in or excluded from a kit of the present disclosure. Individual components of a kit may be formulated together, where appropriate, or be present in separate containers, e.g., to be combined or used separately by the end user.

[0311] In some embodiments, an exemplary kit may be a kit, such as a kit for treating hematological cancers, that comprises a DR-18 polypeptide and a multi-specific antibody, such Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 as a bispecific antibody, such as a bispecific T cell engager (BiTE), such as e.g., blinatumomab. The kit is designed to facilitate the combined administration or co-administration of these therapeutic agents in a prescribed regimen to patients requiring such treatments. The DR-18 polypeptide and the bispecific antibody may be provided in separate containers, where separate containers may provide, e.g., for optimal dosing flexibility and administration sequencing.

[0312] Within the kit, the DR-18 polypeptide is presented in a sterile solution. The DR-18 solution may be formulated in a read-to-use format for direct administration or may present in a concentrated format for dilution into a suitable diluent prior to administration. DR-18 solutions may be formulated suitable for various routes of delivery, such as e.g., subcutaneous, intravenous, or intraperitoneal administration. In some instances, the DR-18 solution may be supplied in a vial or pre-filled syringe, ensuring the stability and shelf-life of the polypeptide.

[0313] Within the kit, the multi- or bispecific antibody is presented in a sterile solution. The multi- or bispecific antibody solution may be formulated in a read-to-use format for direct administration or may present in a concentrated format for dilution into a suitable diluent prior to administration. Multi- or bispecific antibody solutions may be formulated suitable for various routes of delivery, such as e.g., subcutaneous, intravenous, or intraperitoneal administration. In some instances, the multi- or bispecific antibody solution may be supplied in a vial or pre-filled syringe, ensuring the stability and shelf-life of the antibody.

[0314] Both agents in the kit are intended for use within a defined therapeutic protocol. For example, a protocol may be employed where the DR- 18 polypeptide is administered to prime the patient’s immune cells, followed by the administration of the multi- or bispecific antibody to target the primed cells to the cancer, thereby promoting cancer cell death or otherwise inhibiting tumor growth. The separate containers ensure that each agent can be individually dosed and timed according to the treatment plan, enhancing the therapeutic outcome and providing healthcare providers with the flexibility to adjust treatment based on patient response.

[0315] Instructions for use, including recommended dosing schedules and administration techniques, may be included in the kit. These instructions are intended to guide healthcare providers in the proper preparation and delivery of the therapeutic agents, ensuring safety and efficacy in treating cancers as prescribed. In this example, the kit is designed to support the combined therapeutic approach of DR-18 and multi-specific antibodies, such as bispecific antibodies, thereby enhancing immune-mediated cancer therapy. Attorney Docket No.: S199207 1060WO (00093)

[0316] ST-020-W01

[0317] Sequences

[0318] TABLE 2 Attorney Docket No.: S199207 1060WO (00093)

[0319] ST-020-W01 Attorney Docket No.: S199207 1060WO (00093)

[0320] ST-020-W01 Attorney Docket No.: S199207 1060WO (00093)

[0321] ST-020-W01 Attorney Docket No.: S199207 1060WO (00093)

[0322] ST-020-W01

[0323] Nonlimiting Embodiments

[0324] Notwithstanding the appended claims, the present disclosure is also defined by the following embodiments:

[0325] 1. A method of treating a subject having a hematological cancer and comprising an inactive anti-cancer immune cell population, the method comprising: (a). identifying, or having identified, a subject having a hematological cancer that is: (i). relapsed and / or refractory to at least one standard of care (SOC) therapy for the hematological cancer; and (ii). renders the subject deficient in the production of anti-drug antibodies;

[0326] (b). administering a plurality of doses of a decoy-resistant interleukin 18 (DR-18) to the identified subject to provide an amount effective to activate the inactive anti-cancer immune cell population against the hematological cancer, thereby treating the subject.

[0327] 2. The method of embodiment 1 , wherein the hematological cancer is a lymphoma and the lymphoma is relapsed and / or refractory to at least one SOC therapy.

[0328] 3. The method of embodiment 1 , wherein the hematological cancer is a leukemia and the leukemia is relapsed and / or refractory to at least one SOC therapy.

[0329] 4. The method of embodiment 1 , wherein the hematological cancer is multiple myeloma and the multiple myeloma is relapsed and / or refractory to at least one SOC therapy.

[0330] 5. The method of any one of embodiments 1-4, further comprising measuring, in a sample from the subject, an amount of B cells present in the sample, an amount of plasma cells present in the sample, or a combination thereof.

[0331] 6. The method of any one of embodiments 1-5, wherein each dose of the plurality of doses is separated by at least five days and no more than nine days. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0332] 7. The method of any one of embodiments 1-6, wherein the method further comprises administering to the subject a multi-specific therapeutic, wherein the multi-specific therapeutic specifically binds to an antigen expressed on cells of the hematological cancer and an antigen expressed on cells of the anti-cancer immune cell population.

[0333] 8. The method of embodiment 7, wherein the multi-specific therapeutic is a bispecific therapeutic or a trispecific therapeutic.

[0334] 9. The method of embodiment 7 or 8, wherein the multi-specific therapeutic is administered after the first dose of the DR-18, optionally at least 24 hours after, optionally at least one week after.

[0335] 10. The method of any one of embodiments 7-9, wherein the hematological cancer is a B cell cancer and the antigen expressed on cells of the hematological cancer is selected from: CD19, CD20, BCMA, CD123, CD38, CD33, FcRH5, and GPRC5D.

[0336] 11. The method of any one of embodiments 7-10, wherein the multi-specific therapeutic is selected from: blinatumomab, mosunetuzumab, epcoritamab, teclistamab, flotetuzumab, talquetamab, and AMG 330.

[0337] 12. The method of any one of embodiments 11 , wherein the multi-specific therapeutic is blinatumomab.

[0338] 13. The method of any one of embodiments 1-12, wherein the method does not comprise an autologous cell transplant or an allogeneic cell transplant.

[0339] 14. The method of any one of embodiments 1-13, wherein each dose of the plurality of doses is an amount from 20 pg / kg to 1200 pg / kg.

[0340] 15. The method of any one of embodiments 1-14, wherein the DR-18 has less than 100% sequence identity and at least 85% sequence identity to WT human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO:64), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H. The method of embodiment 15, wherein the DR-18 comprises an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62 and 63, optionally wherein the DR IL-18 polypeptide consists of, or consists essentially of, an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62 and 63. The method of any one of embodiments 1-16, wherein each dose of the plurality of doses is from 30 pg / kg to 1200 pg / kg, optionally from 60 pg / kg to 240 pg / kg. A method treating a subject having a cancer, the method comprising treating the subject with a combination therapy comprising administering to the subject a plurality of doses of a decoy-resistant interleukin 18 (DR- 18) and a multi-specific therapeutic, wherein the multi-specific therapeutic specifically binds to an antigen expressed on cells of the cancer and an antigen expressed on immune cells within the subject. The method of embodiment 18, wherein the multi-specific therapeutic is a trispecific therapeutic or a bispecific therapeutic. The method of any one of embodiments 18-19, wherein the antigen expressed on immune cells is a CD3 antigen. The method of any one of embodiments 18-20, wherein the antigen expressed by cells of the cancer is selected from CD19, EpCAM, CD20, GP100, BCMA, EGFR, cMET, HER2, and HER3. The method of embodiment 21, wherein the antigen expressed by cells of the cancer is selected from CD19, EpCAM, CD20, GP100, and BCMA. The method of embodiment 22, wherein the antigen expressed by cells of the cancer is selected from CD19, CD20, and BCMA. The method of embodiment 23, wherein the antigen expressed by cells of the cancer is CD19. The method of embodiment 23, wherein the antigen expressed by cells of the cancer is CD20. The method of embodiment 23, wherein the antigen expressed by cells of the cancer is BCMA. The method of any one of embodiments 18-26, wherein the bispecific therapeutic is selected from blinatumomab, amivantamab, catumaxomab, mosunetuzumab, epcoritamab, tebentafusp, teclistamab, and zenocutuzumab. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0341] 28. The method of embodiment 27, wherein the bispecific therapeutic is selected from blinatumomab, catumaxomab, mosunetuzumab, epcoritamab, tebentafusp, and teclistamab.

[0342] 29. The method of embodiment 28, wherein the bispecific therapeutic is blinatumomab.

[0343] 30. A kit for treating a cancer in a subject, the kit comprising: a. a decoy- resista nt interleukin 18 (DR-18) polypeptide in a first container; and b. a multi-specific antibody in a second container, wherein the multi-specific antibody specifically binds to an antigen expressed on cells of the cancer and an antigen expressed on immune cells within the subject.

[0344] 31. The kit of embodiment 30, wherein the DR-18 polypeptide is provided in a pre-filled syringe for subcutaneous administration.

[0345] 32. The kit of embodiment 30 or 31 , wherein the multi-specific antibody is a bispecific T cell engager.

[0346] 33. The kit of any one of embodiments 30-32, further comprising an instructional material providing a recommended dosing schedule for the DR-18 polypeptide and the bispecific antibody.

[0347] 34. The kit of any one of embodiments 30-33, wherein the cancer is a hematological cancer.

[0348] 35. The kit of embodiment 34, wherein the hematological cancer is a B cell cancer and the antigen expressed on cells of the hematological cancer is selected from: CD19, CD20, BCMA, CD123, CD38, CD33, FcRH5, and GPRC5D.

[0349] 36. The kit of any one of embodiments 30-35, wherein the DR-18 polypeptide has less than 100% sequence identity and at least 85% sequence identity to wild-type human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO:64), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and the X at pos. 155 is N, K or H. Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0350] 37. The kit of any one of embodiments 29-37, wherein the multi-specific antibody specifically binds to CD19 and CD3 antigens.

[0351] 38. The kit of any one of embodiments 30-37, wherein the multi-specific antibody is blinatumomab.

[0352] Examples

[0353] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

[0354] Example 1 - DR- 18 reactivates immune cells that target cancer

[0355] Immune cell exhaustion represents a significant challenge in cancer immunotherapy. Immune cells, initially effective in targeting and destroying cancer cells, often become functionally impaired in the tumor microenvironment. Prolonged exposure to tumor antigens and the immunosuppressive milieu, characterized by the presence of inhibitory cytokines and regulatory immune cells, leads to a state of exhaustion where the immune cells can display reduced proliferative capacity, decreased cytokine production, and diminished cytotoxic activity. This exhaustion results in an inadequate immune response against the tumor and allows for tumor progression, metastasis, recurrent cancer, and refractory cancer.

[0356] The induction of exhaustion is often marked by the upregulation of inhibitory receptors such as, e.g., PD-1 , CTLA-4, and TIM-3 on T cells, and KLRG1 and NKG2A on NK cells. These receptors play roles in the attenuated signaling pathways that diminish the cells' immune functions. Overcoming T cell and NK cell exhaustion has been a focal point of immunotherapy research, with strategies, including checkpoint inhibitors and adoptive cell transfer, showing promise in restoring immune cell activity. Reversing exhaustion, or reactivating immune cells, is a strategy aimed at rejuvenating the anti-tumor capabilities of a subject’s own immune system (and / or immune cells transferred to a subject), thereby enhancing the efficacy of cancer immunotherapies. Strategies to prevent immune cell exhaustion in order to allow immune cells to continue to control a subject’s cancer, and / or attack a recurrent or refractory cancer, would also be valuable for enhancing treatment efficacy. Attorney Docket No.: S199207 1060WO (00093)

[0357] ST-020-W01

[0358] Effector functions that are progressively lost in exhausted T cells include capacities to proliferate, produce cytokines, and lyse upon chronic antigen exposure. Exhausted T cells are characterized by various phenotypic features, including e.g., upregulation of inhibitory receptors (such as e.g., PD-1 , CTLA-4, LAG-3, TIGIT, and the like), chronic TOR activation, downregulation of activating genes, and gene expression changes in exhaustion-related genes (such as, e.g., inhibitory receptor genes, transcription factors, signaling molecules, chemokine receptors, metabolism genes, and the like). Transcription factor states, collectively or individually, such as e.g., high TCF-1- expression, high T-bet to EOMES ratio, NFAT complexed with AP-1, high PRDM1 expression, low NR4A expression, and low TOX expression, can characterize effector T cells. Whereas these characteristics together with low PRDM1 expression and high BATF expression, collectively or individually, can characterize progenitor exhausted T cells. Transcription factor states, collectively or individually, such as e.g., low TCF- 1- expression, low T-bet to EOMES ratio, high BATF expression, NFAT homodimer, high PRDM1 expression, low NR4A expression, and low TOX expression, can characterize exhausted T cells. See e.g., Jenkins et al. (2023) Oxf Open Immunol. 4(1):iqad006. Further description of T cell exhaustion can be found in Zarour Clin Cancer Res (2016) 22(8): 1856- 1864.

[0359] Regarding NK cells, exhausted NK cells can display various phenotypic characteristics, including e.g., decreased expression of one or more NK cell surface markers often downregulated in the presence of tumors or chronic infection (such as e.g., NKG2D, DAP10, CD16 , NCRs (e.g., NKp30, NKp44, and NKp46), CD226, 2B4, and the like), upregulation of inhibitory receptors (such as e.g., PD-1 , NKG2A, Tim-3, and the like), expression changes in transcriptional programs (e.g., Eomes downregulation, T-bet downregulation, and the like), as well as combinations thereof. See e.g., Bi & Tian, Front. Immunol. (2017) 8(760):1-10.

[0360] We have investigated the ability of DR-18 to reactivate exhausted immune cells. Briefly, immune cells were incubated on tumor cells in culture to induce exhaustion. Markers of cell activation and exhaustion were analyzed to monitor cell state. Exhausted immune cells were further cultured in the presence or absence of DR-18 and further assessed to determine the impact of DR-18 on immune cell exhaustion.

[0361] Materials and Methods

[0362] Ex Vivo Mixed Lymphocyte Reaction (MLR) Assay: CD3+ T cells and CD14+ monocytes were isolated from individual donor peripheral blood mononuclear cells (PBMC). CD14+ monocytes were differentiated by culture with IL-4 and GM-CSF, and then matured via LPS and interferon gamma treatment to generate mature dendritic cells (mDC). CD3+ T cells underwent Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 either single stimulation to generate T effector cells (Teff) or repeat stimulation to generate exhausted T cells (Tex). The resulting cell populations were co-cultured, i.e., mDC + Teff or mDC + Tex, for the MLR assay with or without DR-18. Cell and culture supernatant were analyzed using FACS, cytokine analysis was performed using TR-FRET analysis, proliferation was measured via tritiated-thymidine incorporation assay, and protein levels (e.g., INFg and TNFa) in harvested supernatant were quantified.

[0363] Ex Vivo Rested and Pre-Stimulated NK cell Tumor Cell Killing and Expression Analysis: CD56+ NK cells were isolated from individual donor samples and NK cell cultures were either rested or pre-stimulated with addition of IL-12 (10 ng / mL) and IL-15 (1 ng / mL) into the culture media. The following day, NucRed A549 tumor cells were added to the rested or pre-stimulated NK cell cultures and the co-cultures were incubated with or without DR-18. Cells were imaged every four hours over 72 h using an IncuCyte S3 machine. After 72h, supernatants were harvested in triplicate per condition for cytokine quantification using Luminex and ELISA. The remaining cells were analyzed by flow cytometry for assessment of the markers of interest. Untreated and vehicle negative controls, as well as a constitutively IL-12 / IL-15 stimulated positive control, were also prepared, cultured, collected, and analyzed in parallel.

[0364] Treatment and assessment of C1498 AML mouse models: The C1498 systemic mouse model was prepared as previously described (see e.g., Zhang et.al, (2009) Blood 114(8)).

[0365] DR-18, ST-001 , was administered in 5 doses at 0.32 mg / kg, 5 uL / g, intraperitoneal, biweekly with the final ST-001 dose administered on Day 15. 5-Azacitidine was administered in 3 doses at 8 mg / kg, 10 ul / g, intravenously, with seven days between doses. Venetoclax was administered in 14 doses at 50 mg / kg, 10 ul / g, orally, once a day. Immune cell depletion was performed by conventional methods using antibodies to deplete specific immune cell populations. Multiple approaches and targets have been developed to deplete immune cells effectively, each with its unique mechanisms and applications. One target for antibody-mediated depletion is B cells, with anti-B cell antibodies, such as anti-CD20 antibodies, being used to deplete B cells in murine or humanized animal models. For example, Rituximab treatment can be used to effectively deplete huCD20+ transgenic B cells in a humanized mouse models (see e.g., Zhang et al., Blood (2005) 106 (11): 2205) and antibodies targeting mouse antigens, such as anti-mouse-CD20, can be used for depletion of endogenous immune cells in mouse models (see e.g., Matte-Martone et al. Biol Blood Marrow Transplant (2010) 16(9): 1222-1230; Damsky et al. J Immunother Cancer (2019) 7: 153). Animals were assessed for tumor volume and body weight, then terminal FACS analysis of immune profile was performed. Attorney Docket No.: S199207 1060WO (00093)

[0366] ST-020-W01

[0367] Results

[0368] Resulting analysis of ex vivo T cell activation using an MLR assay showed that, despite donor-to-donor variability, DR-18 has the ability to rescue proliferation and activation of exhausted T cells.

[0369] DR-18 shows robust single agent activity in C1498 mouse model of acute myeloid leukemia (AML). For example, a mouse DR-18 surrogate of the Test Compound described below (referred to herein as ST-001) substantially inhibited C1498 tumor growth when administered to mice as a monotherapy, even when compared to other standard therapies, such as standard of care (SOC) Azacytidine and Venetoclax regimen ("Aza+VentoClax") (FIG. 1). DR-18 administration also improved survival whether administered as a monotherapy or in combination with SOC Azacytidine / Venetoclax regimen (FIG. 2). Average percentage increase in life span, as compared to vehicle negative control, observed were 27.8%, 33.3%, and 61.1% for the ST-001 monotherapy, Aza+VentoClax therapy, and ST-001 + Aza+VentoClax therapy, respectively (p-value < 0.001).

[0370] Further studies were performed to identify the active component of the immune system in tumor reduction observed following administration of DR-18 in the C1498 model. Depletion studies were performed where various immune cell populations, such a CD4+, CD8+, NK1.1+ or CD4+ / CD8+ cell populations, were depleted and the resulting impact on DR-18 tumor reduction was assessed. As shown in FIG. 3, mice treated with ST-001 and CD4+ T cell depletion showed similar tumor growth inhibition as compared to ST-001 monotherapy without immune cell depletion (compare "ST-001" with "anti-CD4"), however, mice treated with ST-001 and CD8+ or CD4+ / CD8+ T cell depletion showed substantially reduced tumor inhibition (compare "ST-001 with "anti-CD8" and "anti-CD4 / 8"). These findings identify CD8+ T cells as an active component of the immune system driving DR-18-mediated tumor reduction in the C1498 model of AML. Based on these depletion experiments, the role of NK cells in tumor growth inhibition in this model was observed to be minor. This is in comparison to, e.g., the significant role NK cells were found to play in DR-18-mediated tumor regression in YUMMER1.7 melanoma models that were MHC class I deficient (see e.g., Zhou et al. Nature (2020) 583(7817):609-614). Without being bound by theory, the differing contributions of NK cells in this instance could be tumor-type specific, dependent upon MHC class I expression status, or some combination thereof.

[0371] Collectively, these results indicate that T cells are capable of being activated by DR- 18 and these cells can play a significant role in the anti-cancer response attributed to DR-18 Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 treatment in hematological cancers, which response can rival or surpass the effectiveness of SOC therapies in these cancers.

[0372] Example 2 - Subject immune responses to administered Decoy-Resistant Interleukin 18 (DR- 18)

[0373] The human cytokine interleukin 18 (IL-18) has been previously investigated for the treatment of cancer in human clinical trials as recombinant human IL-18 (rhlL-18). However, clinical development of rhlL-18 has been curtailed by its lack of efficacy due, at least in part, to negative regulation by a decoy receptor called IL-18 binding protein (IL-18BP). Accordingly, engineered versions of IL-18 that maintain IL-18 signaling but are resistant to inhibition by IL- 18BP are under development. In humans, rhlL-18 was found to be safe and well-tolerated.

[0374] Moreover, during clinical testing patient-derived treatment-related antibodies to rhlL-18 were either not detected (see e.g., Robertson et al. J Immunother (2013) 36(6):331-34) or found to not impact toxicity, pharmacokinetic, or pharmacodynamic variables (see e.g., Robertson et al., Clin Cancer Res (2008) 14(11):3462-69). However, patient responses to rhlL-18 are not necessarily predictive of patient responses to engineered decoy-resistant versions. Given a high degree of amino acid sequence similarity between wild-type human IL-18 and human DR-18's (e.g., >90% sequence identity), similar immunogenicity may be expected. Nevertheless, the precise human biological responses to engineered versions of IL-18, and particularly any antidrug antibody (ADA) and / or neutralizing antibody (nAb) responses, were unpredictable.

[0375] Non-human primate (NHP) responses to a DR-18

[0376] A Test Compound, that is a DR- 18, has demonstrated considerably improved efficacy compared to wild-type recombinant IL-18 in preclinical syngeneic mouse models both as monotherapy and in combination with checkpoint inhibitors, including in immunogenic (“hot”) and checkpoint inhibitor resistant (“cold”) tumors.

[0377] The Test Compound has been studied in in vivo pharmacokinetic (PK) studies in non- human primates (NHPs). These studies were performed, in part, to provide information on the nonclinical pharmacokinetics / toxicokinetics of the Test Compound.

[0378] A 4-Week toxicity study of the Test Compound following subcutaneous (SC) and intravenous (IV) injection in cynomolgus monkeys with a 2- week recovery period was performed. The objectives of the study were to evaluate the potential toxicity of the Test Compound and determine the toxicokinetic characteristics of the Test Compound when administered once weekly on Days 1 , 8, 15, and 22, and to evaluate the potential reversibility of any findings following a 2-week recovery period. Attorney Docket No.: S199207 1060WO (00093)

[0379] ST-020-W01

[0380] Test Compound at 30 mg / mL was used to prepare dose formulations and formulations were filtered using a 0.2 pm polyvinylidene fluoride (PVDF) filter prior to dose administration. Purpose-bred naive cynomolgus monkeys (Macaca fascicularis) of 2 to 4 years of age and weighing 1.8 to 4.0 kg at initiation dosing were employed.

[0381] TABLE 3

[0382] The dose formulations were administered to appropriate animals in Group 1 by subcutaneous injection into the interscapular area once weekly on Days 1 , 8, 15, and 22. On each dosing day, the SC administration in Group 1 animals was followed by an intravenous infusion over 60 minutes (±6 minutes) starting within 5 minutes of completion of the SC dose. Dose formulation was administered using a temporary catheter inserted into a peripheral vein connected to a primed infusion line. The appropriate volume was delivered using an infusion pump.

[0383] The dose formulations were administered to appropriate animals in Groups 2 to 4 by subcutaneous injection into the interscapular area once weekly on Days 1 , 8, 15, and 22.

[0384] The dose formulations were administered to appropriate animals in Group 5 by intravenous infusion over 60 minutes (±6 minutes) once weekly on Days 1 , 8, 15, and 22. Dose formulation was administered using a temporary catheter inserted into a peripheral vein connected to a primed infusion line. The appropriate volume was delivered using an infusion pump.

[0385] Dose administration was completed within 6 hours from the removal of formulations from the refrigerator and within 90 hours from the formulation preparation completion time. Various experimental observations and measurements were performed, including mortality / morbidity Attorney Docket No.: S199207 1060WO (00093)

[0386] ST-020-W01 checks, food evaluations, cage side observations, detailed examinations, body weight measurements, ophthalmology, body temperature measurements, and electrocardiography.

[0387] For clinical pathology, animals were fasted for at least four hours before sample collections. Blood samples were collected from an appropriate peripheral vein other than the one used for dosing for up to 24 hours post dosing. Blood samples were collected once from all animals during acclimation and at days 2, 29, and 47 for all animals in groups 1 to 5. Numerous hematology and serum chemistry parameters were assessed.

[0388] For laboratory analyses blood was collected from all animals in groups 1 to 5 on Day 1 at pre-dose and 0.25, 0.5, 1, 1.5, 4, 6, 9, 12, 24, and 48 hours post-dose; on Day 15 pre-dose; and on Day 22 pre-dose and 0.25, 0.5, 1 , 1.5, 4, 6, 9, 12, 24, and 48 hours post-dose. Postdose for Groups 2 to 4 was following the end of injection and post-dose for Groups 1 and 5 was following the start of infusion. TK sample processing followed standard procedures, including refrigerated centrifugation within 60 minutes of collection, aliquoting, storage, and shipment on dry ice, and testing (and retesting where necessary) by a qualified laboratory. TK parameters were computed for Day 1 and 22.

[0389] For ADA analysis, blood samples were collected once in acclimation and prior to dose administration on Days 1 , 15 and 22. ADA sample processing followed standard procedures, including refrigerated centrifugation within 60 minutes of collection, aliquoting, storage, and shipment on dry ice, and testing (and retesting where necessary) by a qualified laboratory. Plasma samples were assayed for the Test Compound and ADA using validated ligand binding assays (LBA) bioanalytical methods. ADA parameters were computed for Day 1 and 22.

[0390] A profound treatment-related anti-drug antibody (ADA) reaction to the Test Compound was observed in NHPs. All animals treated with the Test Compound tested positive to ADAs on Day 15 and / or Day 22 collections. Without being bound by theory, the presence of ADAs likely impacts the measured PK / TK parameters of the Test Compound. For example, the presence of ADAs appeared to impact TK parameters estimation in all animals administered the Test Compound subcutaneously, as evidenced by a steeper decline of the terminal elimination phase and increased clearance on Day 22.

[0391] Thus, in contrast to what has been observed in human clinical trials using hrlL-18, the presence of ADA appeared to impact Test Compound toxicokinetic (TK) parameters. A summary of the clearance (CL) parameters for male and female animals following single (Day 1) and repeat (Day 22) dosing is provided in the following table: Attorney Docket No.: S199207 1060WO (00093)

[0392] ST-020-W01

[0393] TABLE 4 The above CL calculations reflect the clearance calculated as Dose / AUCinf , where

[0394] AUCinf is calculated as the area under the plasma concentration time curve extrapolated to infinity, calculated as AUCIast + Clast / AZ , where Clast is the measured concentration at time Tlast (i.e., the time after dosing at which the last quantifiable concentration was observed), AUCIast is the area under the concentration versus time curve from the start of dose administration to the time after dosing at which the last quantifiable concentration was observed, using the linear trapezoidal method, and AZ is the apparent elimination rate constant, estimated by linear regression of the terminal linear portion of the log concentration versus time curve.

[0395] As the data readily shows, CL at Day 22 after repeat dosing was substantially higher than the initial CL parameters calculated on Day 1. Without being bound by theory, such an increase in clearance may be indicative of the high prevalence of ADAs targeting the Test Compound and mediating its clearance during repeat dosing. As drug exposure is dependent on the clearance rate, an ADA facilitated increase in clearance has the potential to reduce overall drug exposure, particularly during repeat dosing. Accordingly, the observed ADA responses Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 profoundly impact the TK profile over time. In fact, the Day 22 TK results show up to 98% reductions in mean Test Compound exposures on Day 22 compared to Day 1 in animals dosed SC (with SC clearance values up to 6280% higher on Day 22 compared to Day 1).

[0396] Further analysis showed that animals administered Test Compound by SC route displayed a markedly lower systemic exposure following repeat dose administration on Day 22, as outlined by the mean accumulation ratios (AR) below 1. These profound differences between Day 1 and Day 22 TK parameters are highlighted, shown in terms of percent change in value (comparing Day 22 to Day 1), in the following table:

[0397] TABLE 5

[0398] Collectively, the above demonstrates a surprising and profound ADA response to an engineered IL-18 in primates. Additionally, an unexpected and substantial impact on TK parameters and drug exposure was observed, characterized by increased drug clearance in the presence of treatment-related ADAs, particularly during repeat dosing.

[0399] Human subject responses to a DR-18

[0400] As part of a Phase 1a and Phase 2 clinical trial of the safety, preliminary efficacy in solid tumors, PK, and PD of a DR18 referred to in this example as "Test Compound", subjects were dosed in a dose escalation monotherapy arm of Test Compound given as a SC injection. Subjects with melanoma, renal cell carcinoma, triple-negative breast cancer, non-small cell lung cancer, squamous cell carcinoma of the head and neck, and microsatellite instability high (MSI- Attorney Docket No.: S199207 1060WO (00093)

[0401] ST-020-W01

[0402] H) solid tumors who have developed disease progression through standard therapy, or for whom SOC therapy that prolongs survival is unavailable or otherwise unsuitable were eligible for the study. The starting dose of Test Compound for dose escalation was 30 pg / kg. All test subjects were pre-medicated for cytokine release syndrome (CRS) prior to dosing with the Test Compound.

[0403] The Test Compound was supplied as a sterile solution for injection to be administered by SC injection. Test subjects initially received weekly treatment with the Test Compound. Additional dosing schedules were available based on emerging safety, tolerability, PK, and preliminary antitumor activity of the Test Compound. Test subjects were treated until one or more various endpoints were reached, including but not limited to, e.g., disease progression, achievement of maximal response, etc., or a decision is made to discontinue treatment.

[0404] The starting dose of the Test Compound was 30 pg / kg, with escalation in the absence of >Grade 2 treatment-related AEs. Subjects were dosed in treatment cycles, with Cycle 1, Day 1 corresponding with the first dose of Test Compound, Cycle 1 , Day 8 corresponding with the second weekly dose, Cycle 1 , Day 15 corresponding with the third dose, Cycle 1 , Day 22 corresponding to the four dose, Cycle 2, Day 1 corresponding with the fifth dose, and so on.

[0405] Samples for ADA and nABs analyses were obtained weekly during Cycle 1 , on Days 1 and 15 of Cycle 2, on Day 1 of Cycle 3, and on Day 1 of every other cycle thereafter, as well as at the end of treatment and at the end of study. Data for ADA and nABs were available for all subjects who received SC Test Compound monotherapy. Of these subjects, about 80% were confirmed to be positive for ADA at some timepoint following SC Test Compound administration: positive ADAs occurred as early as Cycle 2 Day 1 for the 30 pg / kg dose, Cycle 1 Day 22 for the 60 pg / kg dose, Cycle 1 Day 15 for the 120 pg / kg dose, and Cycle 1 Day 22 for the 240 pg / kg dose. Neutralizing ADAs were developed and confirmed in about 40% of subjects and occurred as early as Cycle 3 Day 1 for the 30 pg / kg dose, Cycle 2 Day 15 for the 60 pg / kg dose, Cycle 3 Day 1 for the 120 pg / kg dose, and at later timepoints for the 240 pg / kg dose. Titer levels were significantly increased with duration of treatment across the dose levels and were highly variable between subjects. Limited data for later treatment cycles suggest that subjects with positive nAb status had lower Test Compound exposure.

[0406] Example 3 - CD3e (human) mice and the A20 BCL syngeneic tumor model treated with DR-18, blinatumomab (anti-CD3-CD19 bispecific T cell engager), or the combination thereof Example 1 demonstrates that immune cells, responsible for tumor growth reduction in hematological cancers, can be activated by administration of DR-18. Example 2 demonstrates that subjects that receive DR-18 produce ADAs and nAbs, likely resulting in reduced exposure Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 to DR-18 (and / or less desirable drug PK) limiting drug effectiveness at a given dose. The reduced immune system function in some hematological cancers, including B cell and plasma cell cancers, can reduce the ability of a subject to mount an immune response to administered DR-18, thereby preventing the production of ADAs and / or nAbs. Accordingly, the effectiveness of therapies combining DR-18 with T cell activators were investigated in models of hematological malignancies where subject ADA and / or nAb production was likely to be reduced.

[0407] As an example of this approach, a combination therapy of DR-18 (ST-001) with a bispecific T cell engager (BiTE) targeting CD19 and CD3 (blinatumomab) was investigated in the A20 mouse model of B cell lymphoma (BCL).

[0408] Materials and Methods

[0409] The A20 mouse model of BCL is utilized for the investigation of hematological malignancies, particularly non-Hodgkin lymphoma (NHL). Transgenic mice carrying a human CD3 epsilon polypeptide (i.e. , CD3E or CD3e) transgene are viable and fertile and useful in experiments involving agents that bind to, interact with, and / or signal through human CD3. CD3e is one of four CD3 subunits which associate with the T-cell receptor (TCR) to generate activation signals in T cells, which interaction plays roles in coupling antigen recognition to intracellular signal-transduction pathways and T-cell development. The A20 cell line, originally derived from a spontaneous BCL in BALB / c mice, is regularly employed in mouse BCL models to study tumor growth, immune cell interactions, and the efficacy of therapeutic agents. The A20 cell line is a murine line characterized by surface expression of B cell markers, including CD19, CD20, and IgM. For pharmacological testing with human-targeted antibodies the A20 line has been engineered to express human CD19 (huCD-19) for this study (see, e.g., www(dot)crownbio(dot)com / model-systems / in-vitro / hucell).

[0410] To establish the A20 lymphoma model, such A20 cells were cultured under standard conditions and subsequently harvested during the logarithmic growth phase. Cells were then washed and resuspended in sterile phosphate-buffered saline (PBS) or an appropriate medium at the desired concentration. BCL was induced by subcutaneous injection of the prepared A20 cells into the flank or another suitable location of syngeneic or immunodeficient mice. Commonly, the BALB / c background strain of mice are used due to their compatibility with A20 cells. In this example, CD3e transgenic mice and A20 cells expressing huCD-19 were employed. The usual inoculation involves injecting approximately 1 x 10A6 to 5 x 10A6 A20 cells in a volume of 100 pL PBS per mouse, however, other inoculation schemes can be used.

[0411] A mouse DR-18, known as ST-001 , was administered at 0.32 mg / kg, biweekly. Blinatumomab was administered in ten doses at 0.05 mg / kg, daily. Monotherapies (ST-001 Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 alone and blinatumomab alone) were performed in parallel with the ST-001 plus blinatumomab combination therapy arm. A negative vehicle control group was also included. Tumor volume over time was measured in cubic millimeters (mm3).

[0412] Blinatumomab is a bispecific T cell engager targeting CD19 (target 1) and CD3e (target 2) currently approved for precursor B-cell lymphoblastic leukemia-lymphoma and active in at least non-Hodgkin's lymphoma, diffuse large B cell lymphoma, Richter's syndrome, and Burkitt's lymphoma. The heavy and light chain amino acid sequences of blinatumomab are as follows:

[0413] Heavy Chain 1 - target 1 : QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYN GKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAMDYWGQGTTVTVS S (SEQ ID NO: 67)

[0414] Light Chain 1 - target 1 : DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPP RFSGSGSGTDFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK (SEQ ID NO: 68)

[0415] Heavy Chain 2 - target 2: DIKLQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK FKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQGTTLTVSS (SEQ ID NO: 69)

[0416] Light Chain 2 - target 2: DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFS GSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK (SEQ ID NO: 70)

[0417] Results

[0418] FIG. 4 provides results demonstrating the effectiveness of the ST-001 plus blinatumomab combination therapy in huCD-19 A20 BCL mice. Characteristic of the model, tumor volume increased rapidly in vehicle (i.e., untreated) control mice. In comparison, ST-001 resulted in a 62% reduction in tumor growth when administered alone (compare "Vehicle" and "ST-001"). Blinatumomab monotherapy also demonstrated reduced tumor growth compared to control, with a loss of durability of response occurring around treatment Day 15 (compare "Vehicle" and "blinatumomab"). The observed loss of durability in blinatumomab monotherapy is characteristic of refractory disease.

[0419] Combination therapy with ST-001 and blinatumomab, however, displayed improved tumor growth inhibition as compared to either monotherapy. The combination therapy unexpectedly demonstrated improved durability as compared to blinatumomab alone (compare "ST-001 + Blina" and "blinatumomab"), suppressing the post-Day 15 tumor growth observed in Attorney Docket No.: S199207 1060WO (00093)

[0420] ST-020-W01 mice treated with blinatumomab alone. Overall the combination therapy arm resulted in more than twice as many tumor regressions as observed in the blinatumomab monotherapy arm and nearly twice as many tumor regressions as observed in the ST-001 monotherapy arm. Collectively, these data indicate that the combination of CD3-CD-19 BiTE plus ST-001 was surprisingly more effective in treating BCL than either ST-001 or BiTE alone, resulting in unexpected enhancements in tumor growth inhibition and durability of response as compared to the monotherapies.

[0421] Example 4 - DR-18 activity in re-challenge BCL

[0422] In Example 3, DR-18 administration improved the durability of the treatment effect as compared to treatment without DR-18. The impact of DR-18 administration on longer-term tumor control was further investigated.

[0423] Materials and Methods

[0424] An A20 mouse model of BCL, as described above but without the CD3e transgene, was utilized. Establishment of A20 lymphoma model and inducement of BCL was performed essentially as described above. ST-001 was administered intraperitoneally at 0.32 mg / kg, biweekly. CPX-351 (CPX, a liposomal formulation of cytarabine and daunorubicin) was used as a reference treatment and administered at 20 milligram per kilogram (mpk) daily for four days.

[0425] For re-challenge experiments, mice that achieved a complete response (CR, tumor size of 0 mm3) on ST-001 monotherapy for A20 BCL (left flank injection) were chosen and reinoculated with A20 cells in the opposite (right) flank at 32 days following the start of monotherapy, corresponding to 18 days after the end of initial ST-001 monotherapy treatment (FIG. 5). A control group of naive mice similarly received a right flank injection of A20 cells on the same day as the mice in the re-challenge group. In addition, tumor monitoring was continued in the remaining mice from the monotherapy group that were not chosen for rechallenge (not pictured in FIG. 5 diagram). Tumor volume over time was measured in cubic millimeters (mm3).

[0426] Results

[0427] DR-18 monotherapy was more active than reference CPX SOC treatment in the A20 preclinical model of BCL (FIG. 6), resulting in 60% CR, 40% PR, and no deaths in the ST-001 treated group. DR-18 is well-tolerated in this model with body weights remaining comparable to vehicle treated controls throughout the time course (FIG. 7).

[0428] As described in the methods above and diagramed in FIG. 5, a subset of mice that achieved a CR were chosen for rechallenge with new implantation of A20 cells in the animal’s Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 opposite (right) flank. Rechallenge implantation was performed at 32 days after the start of initial ST-001 treatment. Growth of newly implanted tumors was efficiently inhibited in all rechallenged mice (FIG. 8), demonstrating that ST-001 treatment promotes lasting immunological memory sufficient to control new tumor growth at sites distant from the initial primary tumor implantation site. Control naive mice that were implanted at the same time, with the same A20 cells, as the rechallenge group is provided in FIG. 8 to demonstrate that the A20 cells used in the rechallenge were fully capable of rapidly forming tumors and to serve as a reference for the rate of tumor growth expected in this model.

[0429] Continued monitoring of tumors in responder (but not complete responder) mice that were not chosen for rechallenge revealed that primary A20 tumors eventually regrew in the absence of continued ST-001 therapy. Without being bound by theory, regrowth of primary tumors in this model may be indicative of emergent immune evasion in some small number of A20 cells that remain after cessation of initial ST-001 treatment. This phenomenon supports treatments directed to promoting more complete tumor clearance, including longer-term treatment with ST-001 , combining ST-001 treatment with other tumor-targeting therapies, and combinations thereof.

[0430] Collectively, the data of this example demonstrates that DR-18 treatment effectively inhibits growth of BCL tumors and contributes to longer-term control of tumor growth by promoting immune memory. DR-18’s ability to enhance the function of immune cells capable of controlling tumor growth upon rechallenge further highlights the usefulness of DR-18’s in the recurrent and / or relapsing cancer setting. The regrowth of primary tumors in some DR-18 responder mice was surprising, particularly considering the complete inhibition of secondary tumor growth observed in complete responder mice that were rechallenged. Whereas the inhibition of secondary tumor growth in rechallenged CR mice demonstrates effective longer- term control, this finding also supports the use of long-term DR-18 treatment schedules and therapies that combine DR-18 treatment with other cancer therapeutics to achieve prolonged control of primary tumors.

[0431] Example 5 - CD3e (human) mice and the MC38-huCLDN18.2 tumor model treated with DR-18, gresonitamab (anti-CD3-CLDN18.2 bispecific T cell engager), or the combination thereof

[0432] Example 3 demonstrates the effectiveness of employing a DR-18 in combination with blinatumomab, a bispecific T cell engager, for the treatment of cancer and Example 4 supports the further development of combination therapies employing DR-18. In this example, the effectiveness of therapies combining DR-18 with T cell activators was further investigated in a Attorney Docket No.: S199207 1060WO (00093)

[0433] ST-020-W01 murine colon adenocarcinoma solid tumor model using ST-001 in combination with gresonitamab, a half-life extended bispecific T-cell engager (BiTE) antibody targeting CD3 and Claudin 18.2 (CLDN18.2).

[0434] Materials and Methods

[0435] The human CD3e transgenic mouse model and its use in syngeneic tumor models to investigate the activity of immune cell targeting therapeutics is described above. The MC38 cell line is a mouse adenocarcinoma colon cancer line derived from a female C57BL / 6 mouse. MC38 can recapitulate features of human colorectal cancer and is useful for elucidating antitumor T cell responses and investigating mechanisms of immunotherapeutics (Shields et al. (2023) Front Immunol. 14:1152035). For pharmacological testing with human-targeted antibodies the MC38 line has been engineered to express human CLDN18.2 (huCLDN18.2) for this study (Crown Bioscience; Available Syngeneic Cell Lines; CLDN18.2).

[0436] To establish the model, such MC38-huCLDN18.2 cells were cultured under standard conditions and subsequently harvested during the logarithmic growth phase. Cells were then washed, quantitated, and resuspended in an appropriate medium at the desired concentration such that each mouse was inoculated with 5 million tumor cells in the right rear flank. Mice were randomized prior to inoculation. In this example, CD3e transgenic mice and MC38 cells expressing huCLDN18.2 were employed.

[0437] ST-001 (also referred to in this example as “DR-18”), was administered at 0.32 mg / kg, in a total of five doses over 2-3 weeks. Five doses of gresonitamab were administered at either 0.05 mg / kg or 0.5 mg / kg also over 2-3 weeks. Monotherapy (DR-18 alone and gresonitamab alone) were each performed in parallel with each DR-18 plus gresonitamab combination therapy arm. A negative “vehicle” (PBS) control group was also included. T umor volume over time was measured in cubic millimeters (mm3).

[0438] Gresonitamab (also referred to as AMG 910) is a bispecific T cell engager targeting human CLDN18.2 (target 1) and human CD3e (target 2). Gresonitamab was in clinical development, including in at least clinical trial NCT04260191 - Study of AMG 910 in Subjects With CLDN18.2-Positive Gastric and Gastroesophageal Junction Adenocarcinoma, but is not currently approved by the U.S. Food and Drug Administration (FDA). Other therapeutic antibodies targeting CLDN18.2, such as zolbetuximab-clzb, have been approved by the FDA and numerous other antibodies targeting CLDN18.2 (see e.g., Nakayama et al. (2024) NatRevCIinOncol 21(5):354-369) either are currently or have recently been in active clinical development, including in at least the agents employed in clinical trials NCT06921928, NCT04671875, NCT05190575, NCT05980416, NCT06005493, NCT04396821 , NCT06792435, Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0439] NCT05639153, NCT05857332, NCT06350006, NCT05934331 , NCT06027346, NCT06219941 , NCT05001516, NCT04495296, NCT06346392, NCT05065710, NCT04914117, NCT06921837, NCT04856150, and NCT05482893. Antibodies targeting CLDN18.2 have been or are being investigated for the treatment of various cancers, including but not limited to, Cldn18.2-positive Advanced Solid Tumors, Biliary Tract Cancer, Colorectal Cancer (inc. metastatic), Digestive System Neoplasms, Gastrointestinal Neoplasms, Esophageal Adenocarcinoma, Gastric Cancer (inc. Gastric Cancer Adenocarcinoma (inc. metastatic) and Gastric Carcinoma), Gastroesophageal Adenocarcinoma, Gastric or Gastroesophageal Junction Adenocarcinoma, Gastroesophageal Junction Cancer, Lung Cancer (NSCLC), Ovarian Cancer, Pancreatic Cancer (inc. Pancreatic Adenocarcinoma (metastatic) and Pancreatic Ductal Adenocarcinoma), and Stomach Neoplasm.

[0440] The heavy and light chain amino acid sequences of gresonitamab are as follows: Heavy Chain 1 : QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQCLEWMGWINPNSGGTKY AQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDRITVAGTYYYYGMDVWGQGTTVT VSS (SEQ ID NO: 71)

[0441] Light Chain 1 : DIQMTQSPSSVSASVGDRVTITCRASQGVNNWLAWYQQKPGKAPKLLIYTASSLQSGVPSRFS GSGSGTDFTLTIRSLQPEDFATYYCQQANSFPITFGCGTRLEIK (SEQ ID NO: 72)

[0442] Heavy Chain 2: EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATY YADSVKDRFTISRDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYWAYWGQGTLVTV SS (SEQ ID NO: 73)

[0443] Light Chain 2: QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPA RFSGSLLGGKAALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL (SEQ ID NO: 74)

[0444] Results

[0445] FIG. 9 provides results demonstrating the effectiveness of the DR-18 plus gresonitamab combination therapy in huCLDN18.2 MC38 colon cancer CD3 mice. Characteristic of the MC38 model, tumor volume increased rapidly in vehicle (i.e., untreated) control mice. In comparison, DR-18 resulted in a reduction in the rate of tumor growth when administered alone (compare "Vehicle" and "DR-18 - 0.32mpk BIW*5"). Gresonitamab monotherapy also demonstrated reduced tumor growth compared to control (compare "Vehicle" and "Greso 0.05mpk BIW*5" and "Greso 0.5mpk BIW*5"). Attorney Docket No.: S199207 1060WO (00093) ST-020-W01

[0446] Combination therapy with DR-18 and gresonitamab, however, displayed improved tumor growth inhibition as compared to either monotherapy. For example, by study day 14, tumors in the DR-18 plus gresonitamab groups were on average less than half the size of tumors in the corresponding gresonitamab only dosage groups (compare e.g., “Greso 0.05mpk BIW*5” vs. “DR-18 + Greso 0.05mpk” and compare, e.g., “Greso 0.5mpk BIW*5” vs. “DR-18 + Greso 0.5mpk”). Addition of gresonitamab also improved efficacy as compared to DR-18 alone (compare, e.g., endpoint tumor size for “DR-18 - 0.32mpk BIW*5” vs. “DR-18 + Greso 0.5mpk”).

[0447] In summary, this MC38 immune reactive syngeneic line expressing human Claudin 18.2 in human CD3e-expressing mice responded to both single agent gresonitamab alone (e.g., 52% tumor growth inhibition (TGI) at 0.05mpk BIW*5 at day 21 ; 79% TGI at 0.5mpk BIW*5 at day 21) and DR-18 alone (89% TGI at 0.32mpk BIW*5 at day 21). However, the combination showed enhanced activity (91 % TGI with lower dose combination at day 21 , 99% TGI with high dose combination at day 21) with increased duration of response. The duration of response, in particular, was surprisingly enhanced by the combination. For example, numerous mice in the higher Greso plus DR-18 dose combination achieved essentially 100% TGI at the end of the 60- day study despite only receiving five doses each of DR-18 and Greso over the initial 2-3 weeks. The combinations were well tolerated (FIG. 11), and 40% of all animals that began the high dose combination displayed sustained CR to day 60 with improved survival as compared to all other conditions (FIG. 10).

[0448] Collectively, these data indicate that the combination of CD3-CLDN18.2 BiTE plus DR- 18 was surprisingly more effective in treating colon cancer than either DR-18 or BiTE alone, resulting in unexpected enhancements in tumor growth inhibition and duration of response as compared to the corresponding monotherapies.

[0449] Example 6 - B16-F10 mouse melanoma cells treated with DR-18, anti-CD3 anti-TRP1 BiTE, or the combination thereof

[0450] B16-F10 is a subline of the murine B16 melanoma cell line which was derived from a spontaneous skin tumor in a mouse. B16-F10 is characterized by low immunogenicity and aggressive metastatic potential. B16-F10 endogenously expresses the antigen Tyrosinase- related protein-1 (TRP1).

[0451] In vivo testing was also performed using the B16-F10 model and anti-CD3 anti-TRP1 BiTE (Absolute Antibody, cat. bAbO136). TRP1 is also known as melanoma antigen gp75; 5,6- dihydroxyindole-2-carboxylic acid oxidase; DHICA oxidase; Catalase B; and Glycoprotein 75. For discussion of the antibody providing the anti-TRP1 binding portions, see Welt et al. (1987) Proc Natl Acad Sci U S A. 84(12):4200-4. The anti-CD3 anti-TRP1 BiTE is a bispecific, knob- Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 into-hole heterodimerized antibody, with one arm directed against TRP-1 (Fab) and one arm directed against mouse CD3e (scFv), mouse lgG2a, kappa.

[0452] Briefly, mice were implanted subcutaneously with B16-F10 cells, harvested in exponential growth phase and quantitated before inoculation (right flank, 2 x 10A5 cells). Mice were randomized and subsequently treated with DR-18 (0.32 mg / kg), anti-CD3 anti-TRP1 BiTE (12.5 pg / kg), or a combination of both (0.32 mg / kg and 12.5 pg / kg, respectively) and tumor growth was monitored by measuring tumor volumes three times per week. A vehicle (i.e., untreated, PBS) control group was also employed in parallel. The body weights of all animals were monitored throughout the study and animals were euthanized if they lost over 20% of their body weight relative to the weight on the day of randomization.

[0453] As shown in FIG. 12, treatment with BiTE showed no to minimal tumor growth inhibition (TGI) in this model, with tumor growth in the BiTE monotherapy group at about 83% of tumor growth observed in vehicle controls. DR-18 alone treatment resulted in about 38% TGI. The DR- 18 and BiTE combination enhanced anti-tumor efficacy as compared to either agent alone, resulting in about 71% TGI, which was greater than the hypothetical additive effects of each agent alone. Moreover, the DR-18 and BiTE combination therapy was well tolerated as no obvious body weight loss was observed during the study period. Wildtype recombinant IL- 18 (WT IL-18) was also tested alone and in combination with anti-CD3 anti-TRP1 BiTE (not pictured). WT IL-18 demonstrated minimal to no single agent efficacy and the combination of WT IL-18 plus anti-CD3 anti-TRP1 BiTE also had minimal to no efficacy in this model.

[0454] Collectively, this example further supports the finding that using DR-18 in a combination treatment with BiTE therapy enhances tumor growth inhibition as compared to the use of either agent alone (and in excess of the hypothetical additive effects of each agent). This example surprisingly demonstrates that the combination of DR-18 and BiTE treatment results in significant tumor grown inhibition when the agents are combined, even when the BiTE is observed to not substantially inhibit tumor growth when used alone and even in a model characterized by low tumor cell immunogenicity. Moreover, B16-F10 is an aggressive tumor line and thus this example also demonstrates the effectiveness, and surprising duration of response, of DR-18 plus BiTE combos in aggressive tumors, and aggressive melanoma in particular.

[0455] In at least some of the previously described embodiments, one or more elements used in an embodiment can interchangeably be used in another embodiment unless such a replacement is not technically feasible. It will be appreciated by those skilled in the art that various other omissions, additions and modifications may be made to the methods and Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 structures described above without departing from the scope of the claimed subject matter. All such modifications and changes are intended to fall within the scope of the subject matter, as defined by the appended claims.

[0456] It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as “open” terms (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases “at least one” and “one or more” to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles “a” or “an” limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “a” or “an” (e.g., “a” and / or “an” should be interpreted to mean “at least one” or “one or more”); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of “two recitations,” without other modifiers, means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to “at least one of A, B, and C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “ a system having at least one of A, B, and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those instances where a convention analogous to “at least one of A, B, or C, etc.” is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., “ a system having at least one of A, B, or C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, Attorney Docket No.: S199207 1060WO (00093) ST-020-W01 either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or “B” or “A and B.”

[0457] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0458] As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1 , 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1 , 2, 3, 4, or 5 articles, and so forth.

[0459] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0460] Accordingly, the preceding merely illustrates the principles of the invention. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. Moreover, nothing disclosed herein is Attorney Docket No.: S199207 1060WO (00093)

[0461] ST-020-W01 intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims.

[0462] The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims. In the claims, 35 U.S.C. §112(f) or 35 U.S.C. §112(6) is expressly defined as being invoked for a limitation in the claim only when the exact phrase "means for" or the exact phrase "step for" is recited at the beginning of such limitation in the claim; if such exact phrase is not used in a limitation in the claim, then 35 U.S.C. § 112 (f) or 35 U.S.C. §112(6) is not invoked.

Claims

1. Attorney Docket No.: S199207 1060WO (00093)ST-020-W01Claims1. A method of treating a subject having a hematological cancer, the method comprising administering a decoy- resista nt interleukin 18 (DR-18) to the subject, thereby treating the subject.

2. A method of priming immune cells of a subject having a hematological cancer, the method comprising administering a decoy-resistant interleukin 18 (DR-18) to the subject, thereby priming the immune cells of the subject.

3. The method of claim 2, further comprising administering to the subject one or more agents that activate the primed immune cells.

4. A method of enhancing cytotoxicity of immune cells in a subject having a hematological cancer, the method comprising administering a decoy-resistant interleukin 18 (DR-18) to the subject, thereby enhancing cytotoxicity of immune cells against the hematological cancer.

5. The method of any one of claims 1-4, wherein the subject has a hematological cancer that is relapsed and / or refractory to at least one standard of care (SOC) therapy.

6. The method of any one of claims 1-5, wherein the subject is deficient in the production of anti-drug antibodies.

7. The method of any one of claims 1-6, wherein the hematological cancer is a lymphoma.

8. The method of any one of claims 1-6, wherein the hematological cancer is a leukemia.

9. The method of any one of claims 1-6, wherein the hematological cancer is multiple myeloma.

10. The method of any one of claims 1-9, wherein the hematological cancer is a B cell cancer.

11. The method of claim 10, wherein the B cell cancer expresses an antigen selected from CD19, CD20, BCMA, CD123, CD38, CD33, FcRH5, and GPRC5D.

12. The method of claim 10 or 11 , further comprising measuring, in a sample obtained from the subject, an amount of B cells present in the sample and / or an amount of plasma cells present in the sample.

13. The method of claim 12, wherein the measuring is performed prior to the administering, optionally wherein the subject is selected for administration of the DR-18 when the measured amount of B cells and / or plasma cells is less than an amount expected in a corresponding healthy subject.Attorney Docket No.: S199207 1060WO (00093)ST-020-W0114. The method of claim 13, wherein the measured amount of B cells is about 10% or less of the total lymphocyte population in the subject, optionally wherein the measured amount of B cells is 200 or less cells per microliter.

15. The method of claim 12, wherein the measuring is performed after the administering for monitoring of the treatment.

16. The method of any one of claims 1-15, wherein the DR-18 has less than 100% sequence identity and at least 85% sequence identity to wild-type human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO:64), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T ; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N ; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and / or the X at pos. 155 is N, K or H.

17. The method of claim 16, wherein the DR-18 comprises an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62 and 63.

18. The method of claim 16 or 17, wherein the DR-18 comprises an amino acid sequence set forth as SEQ ID NO: 20.

19. The method of claim 16, wherein the DR-18 consists of, or consists essentially of, an amino acid sequence selected from SEQ ID NOs: 7, 17-22, 55-57, 62 and 63.

20. The method of claim 19, wherein the DR-18 consists of, or consists essentially of, an amino acid sequence set forth as SEQ ID NO: 20.

21. The method of any one of claims 1-20, wherein the DR-18 is administered to the subject in a plurality of doses.

22. The method of any one of claims 1-21 , wherein a dose of DR-18 or each dose of the plurality of doses is an amount from 20 pg / kg to 1200 pg / kg.Attorney Docket No.: S199207 1060WO (00093)ST-020-W0123. The method of any one of claims 1-21, wherein a dose of DR-18 or each dose of the plurality of doses is in an amount from 30 pg / kg to 1200 pg / kg, optionally from 60 pg / kg to 240 pg / kg.

24. The method of any one of claims 21-23, wherein each dose of the plurality of doses is separated by at least five days and no more than nine days.

25. The method of any one of claims 1-24, wherein the method further comprises administering to the subject a multi-specific therapeutic, wherein the multi-specific therapeutic binds to an antigen expressed on cells of the hematological cancer.

26. The method of claim 25, wherein the multi-specific therapeutic binds to the antigen expressed on cells of the hematological cancer and to an antigen expressed on immune cells in the subject.

27. The method of claim 25 or 26, wherein the multi-specific therapeutic is a bispecific therapeutic.

28. The method of any one of claims 25-27, wherein the multi-specific therapeutic is administered after the first dose of the DR- 18, and optionally at least 24 hours after, optionally at least one week after.

29. The method of any one of claims 25-28, wherein the multi-specific therapeutic is selected from blinatumomab, mosunetuzumab, epcoritamab, teclistamab, flotetuzumab, talquetamab, and AMG 330.

30. The method of claim 29, wherein the multi-specific therapeutic is blinatumomab.

31. The method of any one of claims 1-30, wherein the method does not comprise an autologous cell transplant or an allogeneic cell transplant.

32. The method of any one of claims 1-31, wherein the treating results in a reduction in the amount of cancerous cells in the subject, optionally as measured by a complete blood count, peripheral blood smear, bone marrow biopsy, flow cytometry, or a molecular diagnostic assay.

33. The method of any one of claims 1-32, wherein the treating results in increased cytotoxic killing of hematological cancer cells.

34. The method of any one of claims 1-33, wherein the treating results in amelioration of at least one symptom of the hematological cancer.

35. The method of any one of claims 1-34, wherein the treating results in minimal / measurable residual disease (MRD) negativity.

36. The method of any one of claims 1-35, wherein the treating results in prevention of MRD positivity or sustained MRD negativity.Attorney Docket No.: S199207 1060WO (00093)ST-020-W0137. The method of any one of claims 1-36, wherein the treating results in a partial therapeutic response.

38. The method of any one of claims 1-37, wherein the treating results in complete therapeutic response.

39. The method of any one of claims 1-38, wherein the treating results in an increase in life expectancy of the subject.

40. The method of claim 39, wherein the increase in life expectancy of the subject is an at least 10%, at least 15%, or at least 20% increase in life expectancy of the subject.

41. The method of 39 or 40, wherein the life expectancy of the subject is increased as compared to the subject’s life expectancy if treated according to the standard of care therapy for the cancer.

42. The method of any one of claims 1-41, wherein no standard of care therapy for the hematological cancer is available that prolongs survival.

43. A method of treating a subject having a cancer, the method comprising administering to the subject a decoy-resistant interleukin 18 (DR-18) and a multi-specific therapeutic, wherein the multi-specific therapeutic binds to an antigen expressed on cells of the cancer.

44. The method of claim 43, wherein the multi-specific therapeutic binds to the antigen expressed on cells of the cancer and to an antigen expressed on immune cells in the subject.

45. The method of claim 43 or 44, wherein the multi-specific therapeutic is a bispecific therapeutic.

46. The method of claim 44 or 45, wherein the antigen expressed on immune cells is a CD3 antigen.

47. The method of any one of claims 43-46, wherein the cancer comprises a solid tumor.

48. The method of any one of claims 43-47, wherein the antigen expressed on cells of the cancer is selected from CD19, EpCAM, CD20, GP100, BCMA, EGFR, cMET, HER2, HER3, CLDN18.2, and TRP-1.

49. The method of claim 48, wherein the antigen expressed on cells of the cancer is selected from CD19, EpCAM, CD20, GP100, and BCMA.

50. The method of claim 48, wherein the antigen expressed on cells of the cancer is selected from CD19, CD20, and BCMA.

51. The method of claim 48, wherein the antigen expressed on cells of the cancer is CD19.

52. The method of claim 48, wherein the antigen expressed on cells of the cancer is CD20.Attorney Docket No.: S199207 1060WO (00093)ST-020-W0153. The method of claim 48, wherein the antigen expressed on cells of the cancer is BCMA.

54. The method of claim 48, wherein the antigen expressed on cells of the cancer is CLDN18.2.

55. The method of claim 48, wherein the antigen expressed on cells of the cancer is TRP-1.

56. The method of any one of claims 43-55, wherein the multi-specific therapeutic binds to CD3 x gp100, CD3 x DLL3, CD3 x PRAME, CD3 x STEAP1, CD3 x MUC16, CD3 x DLL3, CD3 x PIWIL1 , CD3E x DLL3 x albumin, CD3 x HER2, CD3 x CLDN18.2, CD3 x CLDN6, CD3 x HLA x MAGEA4, CD3 x PSMA, CD3 x CEA, CD3 x EGFR, CD3 x PRAME, CD3 x EpCAM, CD3 x EGFR, CD3 x PSMA, CD3 x GPC3, CD3 x CDH3 x MSLN, CD3 x CDH17, CD3 x TRP-1, CD3 x NCR3LG1 , CD3 x 5T4, CD3 x CLDN6, CD3 x CLDN6, CD3 x EGFR, CD3 x KLK2, CD3 x ENPP3, CD3 x CALR, CD3 x CEACAM5, CD3 x R0R1, CD3 x EGFRvlll, CD3 x CD3E x HER2, CD3 x 4-1 BB x DLL3, CD3 x CLDN6, CD3 x ENPP3, CD3 x MSLN, CD3 x PSMA, CD3 x MSLN, CD3 x CD20, CD3 x GPRC5D, CD3 x BCMA, CD3 x CD19, CD3 x BCMA, CD3 x CD20, CD3 x CD20, CD3 x CD20, CD3 x BCMA, CD3 x CD19 x CD3D, CD3 x CD123, CD3 x FCRL5, CD3 x BCMA, CD3 x GPRC5D, CD3 x CD123, CD3 x FLT3, CD3 x CD19, CD3 x CD38, CD3 x BCMA x CD38, CD3 x CD22, CD3 x BCMA x GPRC5D, CD3 x CD20 x CD79B, CD3 x CD123, CD3 x PD-1, CD3 x BCMA, CD3 x CD19 x CD2, CD3 x CD38, CD3 x BCMA, CD3 x EpCAM, CD3 x MUC16, CD3 x DLL3, CD3 x GPC3, CD3 x CD19 x CD20 x HER2, CD3 x HER2, CD3 x TAA, CD3 x CD33, CD3 x CD276, CD3 x EpCAM, CD3 x BCMA x GPRC5D, CD3 x CD19 x CD20, CD3 x BCMA, CD3 x CD20, CD3 x CD20, CD3 x BCMA x GPRC5D, CD3 x GPRC5D, CD3 x CD19 x CD28, CD3 x BCMA x GPRC5D, or CD3 x CD276.

57. The method of any one of claims 43-56, wherein the multi-specific therapeutic is selected from blinatumomab, amivantamab, catumaxomab, mosunetuzumab, epcoritamab, tebentafusp, teclistamab, and zenocutuzumab.

58. The method of claim 57, wherein the bispecific therapeutic is selected from blinatumomab, catumaxomab, mosunetuzumab, epcoritamab, tebentafusp, and teclistamab.

59. The method of claim 58, wherein the bispecific therapeutic is blinatumomab.

60. The method of any one of claims 43-56, wherein the multi-specific therapeutic is selected from givastomig, gresonitamab, spevatamig, and tixentamig.

61. The method of claim 60, wherein the multi-specific therapeutic is gresonitamab or tixentamig.Attorney Docket No.: S199207 1060WO (00093)ST-020-W0162. The method of claim 61 , wherein the multi-specific therapeutic is gresonitamab.

63. The method of any one of claims 43-62, wherein the treating results in increased cytotoxic killing of cancer cells.

64. The method of any one of claims 43-63, wherein the treating results in amelioration of at least one symptom of the cancer.

65. The method of any one of claims 43-64, wherein the treating results in minimal / measurable residual disease (MRD) negativity.

66. The method of any one of claims 43-65, wherein the treating results in prevention of MRD positivity or sustained MRD negativity.

67. The method of any one of claims 43-66, wherein the treating results in a partial therapeutic response.

68. The method of any one of claims 43-67, wherein the treating results in complete therapeutic response.

69. The method of any one of claims 43-68, wherein the treating results in an increase in life expectancy of the subject.

70. The method of claim 69, wherein the increase in life expectancy of the subject is an at least 10%, at least 15%, or at least 20% increase in life expectancy of the subject.

71. The method of 69 or 70, wherein the life expectancy of the subject is increased as compared to the subject’s life expectancy if treated according to the standard of care therapy for the cancer.

72. The method of any one of claims 43-71 , wherein no standard of care therapy for the cancer is available that prolongs survival.

73. A kit for treating a cancer in a subject, the kit comprising a decoy-resistant interleukin 18 (DR-18) polypeptide in a container.

74. A kit for treating a cancer in a subject, the kit comprising: a. a decoy-resistant interleukin 18 (DR- 18) polypeptide in a first container; and b. a multi-specific antibody in a second container, wherein the multi-specific antibody binds to an antigen expressed on cells of the cancer.

75. The kit of claim 74, wherein the multi-specific therapeutic binds to the antigen expressed on cells of the cancer and to an antigen expressed on immune cells in the subject.

76. The kit of any one of claims 73-75, wherein the DR-18 polypeptide is provided in a prefilled syringe for subcutaneous administration.

77. The kit of claim of any one of claims 74-76, wherein the multi-specific antibody is a bispecific T cell engager.Attorney Docket No.: S199207 1060WO (00093)ST-020-W0178. The kit of any one of claims 73-77, further comprising an instructional material providing a recommended dosing schedule for the DR-18 polypeptide and / or the bispecific antibody.

79. The kit of any one of claims 73-78, wherein the cancer is a hematological cancer.

80. The kit of claim 79, wherein the hematological cancer is a B cell cancer and the antigen expressed on cells of the hematological cancer is selected from CD19, CD20, BCMA, CD123, CD38, CD33, FcRH5, and GPRC5D.

81. The kit of any one of claims 73-80, wherein the DR-18 polypeptide has less than 100% sequence identity and at least 85% sequence identity to wild-type human IL-18 (SEQ ID NO: 1) and comprises the following amino acid sequence: XFGKXESXLSVIRNLNDQVLFIDQGNRPLFEDMTDSDXRDNAPRTIFIISXYXDXXXRXX AVTISVKXEKISTLSXXNKIISFKEMNPPDNIKDTKSDIIFFXRXVPGHXXKXQFESSSYEG YFLAXEKERDLFKLILKKEDELGDRSIMFTXQXED (SEQ ID NO:64), wherein the X at position (“pos.”) 1 is Y, R or H; the X at pos. 5 is L, H, I or Y; the X at pos. 8 is K, Q or R; the X at pos. 38 is C or S; the X at pos. 51 is M, T, K, D, N, E or R; the X at pos. 53 is K, R, G, S or T; the X at pos. 55 is S, K or R; the X at pos. 56 is Q, E, A, R, V, G, K, L or R; the X at pos. 57 is P, L, G, A or K; the X at pos. 59 is G, A or T; the X at pos. 60 is M, K, Q, R or L; the X at pos. 68 is C, S, G, A, V, D, E or N; the X at pos. 76 is C or S; the X at pos. 77 is E or D; the X at pos. 103 is Q, E, K, P, A or R; the X at pos. 105 is S, D, N, R, K or A; the X at pos. 110 is D, K, H, N, Q, E, S or G; the X at pos. 111 is N, H, Y, D, R, S or G; the X at pos. 113 is M, V, R, T or K; the X at pos. 127 is C or S; the X at pos. 153 is V, I, T or A; and / or the X at pos. 155 is N, K or H.

82. The kit of claim 81, wherein the DR-18 polypeptide comprises an amino acid sequence set forth as SEQ ID NO: 20.

83. The method of claim 81 or 82, wherein the DR-18 polypeptide consists of, or consists essentially of, an amino acid sequence set forth as SEQ ID NO: 20.

84. The kit of any one of claims 73-83, wherein the multi-specific antibody binds to CD19 and CD3 antigens, CLDN18.2 and CD3 antigens, or TRP-1 and CD3 antigens.

85. The kit of any one of claims 73-83, wherein the multi-specific antibody is blinatumomab.

86. The kit of any one of claims 73-83, wherein the multi-specific antibody is gresonitamab.

Images