Compositions, systems and methods for treating cancer using tumor therapy electric fields in combination with immune checkpoint inhibitors and MHC class I activators

Abstract

Compositions, systems, and methods are disclosed for reducing the viability of cancer cells and treating cancer, as well as preventing an increase in volume of a tumor present in the body of a living subject. The systems and methods involve applying an alternating field while administering at least one composition that increases the expression of MHC class I molecules in cancer cells.

Description

[0001] Cross-reference to related applications

[0002] Pursuant to 35 U.S.C., 119(e), this application claims the benefit of U.S. Provisional Application No. 63 / 584,254, filed September 21, 2023. The entire contents of the aforementioned patent application are hereby expressly incorporated herein by reference.

[0003] Statement on Federally Funded Research

[0004] not applicable. Background Technology

[0005] Tumor therapeutic electric fields (TTFields) are low-intensity (e.g., 1 V / cm to 3 V / cm) alternating electric fields in the mid-frequency range (such as, but not limited to, 100 kHz to 500 kHz) that target solid tumors via mitosis. This non-invasive treatment of targeted solid tumors is described, for example, in U.S. Patent Nos. 7,016,725; 7,089,054; 7,333,852; 7,565,205; 8,244,345; 8,715,203; 8,764,675; 10,188,851; and 10,441,776. TTFields are typically delivered via two pairs of transducer arrays that generate a vertical electric field within the treated tumor; the electrode arrays constituting each of these pairs are positioned on opposite sides of the body site being treated. More specifically, for the OPTUNE® system, one pair of electrodes is positioned to the left and right of the tumor (LR), and another pair of electrodes is positioned to the front and back of the tumor (AP). TTFields is approved for the treatment of glioblastoma multiforme (GBM) and can be delivered, for example, via the OPTUNE® system (Novocure Limited, St. Helier, Jersey), which includes a transducer array placed on the patient's laser head.

[0006] Each transducer array used to deliver TTFields in the OPTUNE® device comprises a set of ceramic disc electrodes coupled to the patient's skin (such as, but not limited to, the patient's shaved head for treating GBM) via a layer of conductive medical gel. The medical gel is designed to deform to conform to the body's contours and provide good electrical contact between the array and the skin; thus, the gel interface bridges the skin and reduces interference. The device is intended to be worn continuously by the patient for 2 to 4 days, then removed for hygiene and regrowth (if necessary), followed by reapplication of a new array. Thus, the medical gel maintains substantially continuous contact with the patient's skin area for each contact period of 2 to 4 days. Furthermore, the array can be moved a few centimeters in either direction to allow the skin to heal between treatment periods. Therefore, when the replaced electrodes are moved slightly, the portion of skin covered by the electrodes / gel for 2 to 4 days is exposed for 2 to 4 days; the device can then be reapplied to the original portion of the skin for the next 2 to 4 days.

[0007] TTFields has been studied in clinical trials in combination with standard of care (SOC) immunotherapy or chemotherapy regimens for the treatment of non-small cell lung cancer that has progressed after platinum-based therapy (Leal et al., (2023), Lancet Oncology, 24(9):1002–1017), and in preclinical studies in combination with immune checkpoint inhibitors (Voloshin et al., (2020), Cancer Immunol Immunother, 69(7):1191–1204; Barsheshet et al., (2022), International Journal of Molecular Sciences, 23(22):14073).

[0008] Downregulation of MHC class I molecules has been observed in 40% to 90% of human tumors, and this is often associated with poor prognosis. Different regulatory mechanisms can be used to reduce the surface presentation of MHC class I molecules in both adult and pediatric tumors. Downregulation of MHC class I molecules (key factors in the initiation of adaptive immune responses) allows tumors to avoid the presentation of tumor-associated antigens and T-cell-mediated cytotoxicity by downregulating the surface presentation of major histocompatibility complex (MHC) class I molecules (Cornel et al., (2020) Cancer (Basel) 12(7):1760). Downregulation of MHC class I molecules has been described as a mechanism by which cancer patients develop intrinsic and acquired resistance to immunotherapy (Taylor et al., (2022) Frontiers in Immunology 13:844866). Furthermore, in pancreatic cancer, autophagy has been shown to be responsible for the degradation of MHC class I molecules, leading to resistance to immunotherapy and suppression of adaptive antitumor immune responses (Yamamoto et al., Nature, 2020, 581:100–105).

[0009] In terms of treatment, increasing the expression of MHC class I molecules has been shown to enhance the efficacy of immune checkpoint blockade (Gu et al., Cancer Discov, 11(6):1524-1541). Furthermore, the immunomodulatory properties of PI3K / AKT / mTOR and MAPK / MEK / ERK inhibition have been shown to enhance the response of melanoma and triple-negative breast cancer to immune checkpoint blockade (Zhang et al., Int J Mol Sci., 23(13):7353, 2022). Moreover, in 2020, the U.S. Food and Drug Administration (FDA) approved the combination of the PD-L1 inhibitor atezolizumab with the BRAF inhibitor vemurafenib and the MEK inhibitor cobimetinib for first-line treatment of patients with advanced melanoma harboring the BRAF V600 mutation. However, in patients with EGFR-mutant non-small cell lung cancer (NSCLC), the combination of the PD-L1 inhibitor durvalumab with the epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) osimertinib did not yield clinical benefit and only resulted in more adverse events (Li et al., Cancer (2023), 15(10):2858).

[0010] TTFields have been shown to induce autophagy (Shteingauz et al., Cell Death Dis, 9(11):1074; Davidi et al., Cancer (Basel), 14(12):2959). Furthermore, autophagy has been shown to contribute to the degradation of MHC class I molecules.

[0011] Autophagy inhibition has been clinically tested in the following trials: hydroxychloroquine in breast cancer patients and chloroquine in stage IV small cell lung cancer (SCLC) patients (US NLM clinical trial identifiers NCT01292408 and NCT00969306, respectively); however, both trials were terminated due to unfavorable results. Furthermore, interferon-γ has been tested as a cancer therapy but without success (Zhang et al., Cancer Immunol Res, 7(8):1237-1243, 2019). In addition, HDAC inhibitors are currently undergoing clinical trials but have been shown to have serious side effects (Subramanian et al., Pharmaceuticals (Basel), 3(9):2751–2767, 2010; and Bondarev et al., British Journal of Clinical Pharmacology, 87(12):4577-4597, 2021).

[0012] Silgner et al. (Neuro-Oncology (2018), 20(6):vi133) investigated the interaction between TTFields and molecules involved in immune responses and drugs that may regulate immune cell activity, and found that increased expression of NKG2D ligand and enhanced NK cell-based killing of glioma cells exposed to TTFields, while the expression of MHC class I and MHC class II molecules remained unchanged. Attached Figure Description

[0013] Figure 1 The results of the analysis of MHC class I molecule expression in human and mouse pancreatic cancer cell lines after 72 or 96 hours of TTFields exposure were graphically depicted (1A-AsPc1 human pancreatic cancer cell line; 1B-Panc02 mouse pancreatic cancer cell line).

[0014] Figure 2Transcriptomic analyses performed on glioblastoma (GBM) cell lines (GIN31, KNS 42, U87, GBM1, GBM, JHH-520, NCH466, SF188), lung adenocarcinoma cell lines (H-1650, HCC4006), ovarian cancer cell lines (OVCAR3), gastric cancer cell lines (AGS, KATOIII), malignant pleural mesothelioma cell lines (MPM), and pancreatic cancer cell lines (AsPC1 and BxPC3) were graphically depicted to identify genes with differentially expressed levels after TTFields treatment compared to control groups.

[0015] Figure 3 The effects of TTFields application on MHC-I expression in Panc2 cells were graphically illustrated, showing a decrease in MHC-I expression followed by an increase in MHC-I expression after simultaneous chloroquine treatment. Panc02-luc mouse pancreatic cancer cells (10,000 cells per well) were incubated for 72 h under the following conditions: untreated control group, 10 μM chloroquine (autophagy inhibitor) group, TTFields (1.75 V / cm and 150 kHz) group, and TTFields combined with 10 μM chloroquine group.

[0016] Figure 4 The effects of TTFields treatment on MHC-I expression in A2780 cells were graphically illustrated, showing a decrease in MHC-I expression following TTFields application and an increase in MHC-I expression following apeleliximab treatment. A2780 human ovarian cancer cells (20,000 cells per well) were incubated for 144 h under the following conditions: untreated control group; 250 nM apeleliximab (PI3K inhibitor) treatment for 72 h: drug-containing medium removed, and fresh medium without apeleliximab added for an additional 72 h incubation; TTFields (1.75 V / cm and 200 kHz) group; TTFields combined with 250 nM apeleliximab treatment for 72 h: drug-containing medium removed, and fresh medium without apeleliximab added for an additional 72 h incubation under TTFields conditions. Detailed Implementation

[0017] Before explaining in detail at least one embodiment of the inventive concept through exemplary language and results, it should be understood that the application of the inventive concept is not limited to the details of the construction and arrangement of the components set forth in the following description. The inventive concept can be implemented in other ways or practiced or performed in various manner. Therefore, the language used herein is intended to give the broadest possible scope and meaning; and these embodiments are intended to be exemplary rather than exhaustive. Furthermore, it should be understood that the wording and terminology used herein are for descriptive purposes and should not be considered limiting.

[0018] Unless otherwise defined herein, scientific and technical terms used in conjunction with the inventive concepts disclosed herein shall have the meanings commonly understood by one of ordinary skill in the art. Furthermore, unless the context requires otherwise, singular terms shall include plural terms, and plural terms shall include singular terms. The foregoing techniques and procedures are generally performed according to conventional methods well known in the art, as described in the various general and more specific references cited and discussed throughout this specification. The terminology, laboratory procedures, and techniques related to analytical chemistry, synthetic organic chemistry, medical chemistry, and medicinal chemistry described herein are well known and commonly used in the art. Standard techniques are used for chemical synthesis and chemical analysis.

[0019] All patents, published patent applications, and non-patent publications mentioned in this specification represent the skill level of a person skilled in the art to which the inventive concept of this disclosure pertains. All patents, published patent applications, and non-patent publications cited in any part of this application are hereby expressly incorporated by reference in their entirety, to the extent that each individual patent or publication is specifically and individually indicated as incorporated by reference.

[0020] All the compositions, components, systems, kits, and / or methods disclosed herein can be prepared and performed without excessive experimentation. While the compositions, components, systems, kits, and methods of the inventive concept have been described with reference to specific embodiments, it will be apparent to those skilled in the art that variations may be made to the compositions and / or methods and steps or sequences of steps described herein without departing from the concept, spirit, and scope of the inventive concept. All such similar substitutions and modifications that are apparent to those skilled in the art are considered to be within the spirit, scope, and concept of the inventive concept as defined by the appended claims.

[0021] As used in accordance with this disclosure, unless otherwise specified, the following terms shall be understood to have the following meanings:

[0022] When used in conjunction with the term "comprising" in the claims and / or description, the use of the terms "a" or "an" can mean "one," but also aligns with the meanings of "one or more," "at least one," and "one or more." Therefore, unless the context clearly indicates otherwise, the terms "a," "an," and "the" include plural indicators. Thus, for example, reference to "a compound" can refer to one or more compounds, two or more compounds, three or more compounds, four or more compounds, or more quantities of compounds. The term "a plurality" means "two or more."

[0023] The term "at least one" should be understood to include any quantity including one and more than one, including but not limited to 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100, etc. The term "at least one" can be extended to 100 or 1000 or more, depending on the terms it is connected to; furthermore, quantities of 100 / 1000 are not considered limiting, as higher limits can also produce satisfactory results. Additionally, the term "at least one of X, Y, and Z" will be understood to include individual X, individual Y, and individual Z, as well as any combination of X, Y, and Z. The use of ordinal terms (e.g., "first," "second," "third," "fourth," etc.) is solely for the purpose of distinguishing two or more items and does not imply any order or sequence of importance or any order of addition of one item relative to another.

[0024] The term "or" used in the claims is intended to indicate inclusion ("and / or") unless explicitly stated to refer only to alternatives, or unless the alternatives are mutually exclusive. For example, the condition "A or B" can be satisfied by any of the following: A is true (or exists) and B is false (or does not exist); A is false (or does not exist) and B is true (or exists); and both A and B are true (or exist).

[0025] As used herein, any reference to “an embodiment,” “an embodiment,” “some embodiments,” “an example,” “for example,” or “example” means that a particular element, feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment. For example, the phrase “some embodiments” or “an example” appearing in different places in the specification does not necessarily refer to the same embodiment. Furthermore, all references to one or more embodiments or examples should be construed as non-limiting to the claims.

[0026] Throughout this application, the term “about” is used to indicate values ​​including inherent error variations in the composition / device / apparatus, variations in the method used to determine the value, or variations present in research subjects. For example, but not as a limitation, when using the term “about,” the specified value may vary from the specified value by adding or subtracting 20 percent, or 15 percent, or 12 percent, or 11 percent, or 10 percent, or 9 percent, or 8 percent, or 7 percent, or 6 percent, or 5 percent, or 4 percent, or 3 percent, or 2 percent, or 1 percent, as such variations are suitable for performing the disclosed methods and as understood by one of ordinary skill in the art.

[0027] As used in this specification and claims, the terms “comprising” (and any form of inclusion, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of inclusion, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unreferenced elements or method steps.

[0028] As used herein, the term "or combinations thereof" refers to all permutations and combinations of the items preceding the term. For example, "A, B, C, or combinations thereof" is intended to include at least one of the following: A, B, C, AB, AC, BC, or ABC, and, if the order is important in the particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing this example, what is explicitly included are combinations containing repetitions of one or more items or terms, such as BB, AAA, AAB, BBC, AAABCCCC, CBBAAA, CABABB, etc. Those skilled in the art will understand that there is generally no limit to the number of items or terms in any combination unless it is obvious from the context.

[0029] As used herein, the term "substantially" means that the subsequently described event or situation occurs completely or substantially. For example, when relating to a particular event or situation, the term "substantially" means that the subsequently described event or situation occurs for at least 80% of the time, or at least 85% of the time, or at least 90% of the time, or at least 95% of the time. For example, the term "substantially adjacent" can mean that two items are 100% adjacent to each other, or that two items are very close to each other but not 100% adjacent, or that a portion of one of two items is not 100% adjacent to another item but is very close to another item.

[0030] The term “pharmaceutically acceptable” means a compound or composition that is suitable for administration to humans and / or animals without excessive adverse side effects (such as, but not limited to, toxicity, irritation and / or allergic reactions) and in a manner that is commensurate with a reasonable benefit / risk ratio.

[0031] As used herein, the terms “patient” or “subject” include both human and veterinary subjects. “Mammalian” for therapeutic purposes means any animal classified as a mammal, including (but not limited to) humans, livestock and farm animals, non-human primates and any other animal with mammary tissue.

[0032] The term "treatment" refers to therapeutic procedures as well as preventative or preventative measures. Individuals requiring treatment include, but are not limited to, individuals who already have a specific condition / disease / infection and individuals at risk of acquiring a specific condition / disease / infection (e.g., individuals requiring preventative / preventative measures). The term "treatment" also refers to the administration of a drug / element / method to a patient for therapeutic and / or preventative / preventative purposes.

[0033] As used herein, the terms “therapeutic composition” or “pharmaceutical composition” refer to agents that can be administered in vivo to produce therapeutic and / or preventive / protective effects.

[0034] The application of a therapeutically effective or prophylactically effective dose is intended to provide therapeutic benefit in the treatment, prevention, and / or management of a disease, condition, and / or infection. The specific therapeutically effective dose can be readily determined by a physician of general practice and can vary depending on factors known in the art, such as (but not limited to) the type of condition / disease / infection, the patient's medical history and age, the stage of the condition / disease / infection, and the co-administration of other agents.

[0035] The term "effective amount" refers to the amount of a bioactive molecule or its conjugates or derivatives, or a therapeutic agent (e.g., an alternating electric field), sufficient to produce a detectable therapeutic effect without undue adverse side effects (such as, but not limited to, toxicity, irritation, and allergic reactions), and proportionate to a reasonable benefit / risk ratio when used in the manner conceived in this invention. Therapeutic effects may include, for example, but not as a limitation, prevention, inhibition, or reduction of the occurrence of at least one condition, disease, and / or infection. The effective amount for a subject will depend on the type of subject, the subject's body size and health status, the nature and severity of the condition / disease / infection to be treated, the method of administration, the duration of treatment, the nature of concurrent therapies (if any), the specific formulation used, etc. Therefore, it is impossible to predefine an exact effective amount. However, the effective amount for a given situation can be determined by a person skilled in the art using conventional experiments based on the information provided herein.

[0036] As used herein, the term "simultaneous therapy" may be used interchangeably with the terms "combined therapy" and "adjunctive therapy" and will be understood to mean that a patient requiring treatment is treated in combination with the treatment of this disclosure, with or receiving another drug targeting the condition / disease / infection. Such simultaneous therapy may be sequential therapy, wherein the patient is first treated with one treatment regimen / drug composition and then with another treatment regimen / drug composition, or two treatment regimens / drug compositions are administered simultaneously. Furthermore, it should be understood that one administration step (such as, but not limited to, administration of TTFields) may last for a longer period than another administration step (i.e., oral or injectable administration of a substance). In these cases of different administration periods, the term "simultaneous" will be understood to mean that the shorter administration step completely overlaps with the longer administration step. However, the term "simultaneous" will include performing the shorter administration step at any point in time during the longer administration step (e.g., at the beginning, middle, or end of the longer administration step, or any other time period in between), and performing the shorter administration step once or multiple times entirely within the time period of the longer administration step. Therefore, the term "simultaneously" does not require that the two application steps be performed within exactly the same time length.

[0037] As used herein, the term "administration" will be understood to include all routes of administration known in the art, including but not limited to oral, topical, transdermal, parenteral, subcutaneous, intranasal, mucosal, intramuscular, intraperitoneal, intravitreal, and intravenous routes, and includes both local and systemic administration. Furthermore, the compositions of this disclosure (and / or their methods of administration) may be engineered to provide delayed, controlled, or sustained release using formulation techniques well known in the art.

[0038] As used herein, the term “target area” refers to the area containing all or part of the cancer, cancer cells, and / or tumor to be treated.

[0039] Turning now to the inventive concept, this document discloses a concurrent therapy for cancer. This concurrent therapy comprises: the use of an alternating electric field (e.g., TTFields) and at least one immune checkpoint inhibitor compound and at least one compound that enhances the expression of MHC class I molecules (wherein the two compounds may be present in the same composition or different compositions, and may be administered simultaneously or sequentially, in whole or in part). The combination of an alternating electric field (e.g., TTFields) with an immune checkpoint inhibitor and an activator of MHC class I molecule expression provides synergistic results in cancer treatment. Such methods can be performed in vitro or in vivo.

[0040] The inventors were surprised to discover that combining TTFields with drugs that enhance the expression of MHC class I molecules could prevent the degradation of MHC class I molecules and restore or even enhance adaptive immunity. They also found that treating cancer with TTFields, drugs that enhance the expression of MHC class I molecules, and immune checkpoint inhibitors produced a synergistic effect and could overcome tumor resistance to immune checkpoint inhibitors.

[0041] Some non-limiting embodiments of this disclosure relate to a method for reducing the viability of cancer cells. The method includes the steps of: (1) applying an alternating electric field to the cancer cells for a period of time; and (2) administering at least one composition to the cancer cells, wherein the at least one composition comprises at least one immune checkpoint inhibitor compound and at least one compound that enhances the expression of MHC class I molecules.

[0042] Certain additional non-limiting embodiments of this disclosure relate to a method for treating cancer in a subject. The method includes the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; and (2) administering to the subject at least one composition, wherein the at least one composition comprises at least one immune checkpoint inhibitor compound and at least one compound that enhances the expression of MHC class I molecules.

[0043] Certain additional non-limiting embodiments of this disclosure relate to a method for reducing the volume of a tumor present in the body of a living subject, wherein the tumor comprises multiple cancer cells. The method includes the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; and (2) administering to the subject at least one composition, wherein the at least one composition comprises at least one immune checkpoint inhibitor compound and at least one compound that enhances the expression of MHC class I molecules.

[0044] Certain additional non-limiting embodiments of this disclosure relate to a method for preventing the growth of a tumor, wherein the tumor is present in the body of a living subject and contains multiple cancer cells. The method includes the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; and (2) administering to the subject at least one composition, wherein the at least one composition comprises at least one immune checkpoint inhibitor compound and at least one compound that enhances the expression of MHC class I molecules.

[0045] Some additional non-limiting embodiments of this disclosure relate to a method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; and (2) administering to the subject at least one composition, wherein the at least one composition comprises at least one immune checkpoint inhibitor compound and at least one compound that enhances the expression of MHC class I molecules.

[0046] Certain additional non-limiting embodiments of this disclosure relate to a method for reducing the viability of cancer cells, the method comprising the steps of: (1) applying an alternating electric field to the cancer cells for a period of time; (2) administering at least one first composition to the cancer cells, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) administering at least one second composition to the cancer cells, wherein the at least one second composition comprises at least one compound that enhances the expression of MHC class I molecules in the cancer cells.

[0047] Certain additional non-limiting embodiments of this disclosure relate to a method for treating cancer in a subject, the method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; (2) administering to the subject at least one first composition, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) administering to the subject at least one second composition, wherein the at least one second composition comprises at least one compound that enhances the expression of MHC class I molecules in cancer cells in the subject.

[0048] Certain additional non-limiting embodiments of this disclosure relate to a method for reducing the volume of a tumor and / or preventing the tumor from increasing in volume, wherein the tumor is present in the body of a living subject and contains multiple cancer cells, the method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; (2) administering to the subject at least one first composition, wherein the at least one composition comprises at least one immune checkpoint inhibitor; and (3) administering to the subject at least one second composition, wherein the at least one second composition comprises at least one compound that increases the expression of MHC class I molecules in cancer cells in the subject.

[0049] Certain additional non-limiting embodiments of this disclosure relate to a method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; (2) administering to the subject at least one first composition, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) administering to the subject at least one second composition, wherein the at least one second composition comprises at least one compound that enhances the expression of MHC class I molecules in cancer cells in the subject; and wherein steps (2) and (3) are performed simultaneously or wholly or partially sequentially. In a particular (but non-limiting) embodiment, the application of the alternating electric field enhances the toxicity of the at least one first composition and / or the at least one second composition to cancer cells in the subject compared to administering the at least one first composition and / or the at least one second composition to the subject without the application of an alternating electric field.

[0050] Certain additional non-limiting embodiments of this disclosure relate to a composition for use in any of the methods disclosed above, which enhances the expression of MHC class I molecules in cancer cells in a subject.

[0051] Certain additional non-limiting embodiments of this disclosure relate to a kit for reducing the viability of cancer cells (or for performing any of the methods described above), the kit comprising: a composition that enhances the expression of MHC class I molecules in cancer cells in a subject; and an electric field generating device configured to apply an alternating electric field to the cancer cells for a period of time.

[0052] Certain additional non-limiting embodiments of this disclosure relate to a system for use in any of the methods disclosed above, the system comprising: a first composition comprising at least one immune checkpoint inhibitor; and a second composition that enhances the expression of MHC class I molecules in cancer cells in the subject.

[0053] Certain additional non-limiting embodiments of this disclosure relate to a kit for reducing the viability of cancer cells (or for performing any of the methods described above), the kit comprising: a first composition comprising at least one immune checkpoint inhibitor; a second composition that enhances the expression of MHC class I molecules in cancer cells in the subject; and an electric field generating device configured to apply an alternating electric field to the cancer cells for a period of time.

[0054] Certain additional non-limiting embodiments of this disclosure relate to a system for reducing the viability of cancer cells (or for performing any of the methods described above), the system comprising: a first composition comprising at least one immune checkpoint inhibitor; a second composition that enhances the expression of MHC class I molecules in cancer cells in the subject; and an electric field generating device configured to apply an alternating electric field to the cancer cells for a period of time.

[0055] The steps of any of the methods disclosed herein may be performed in combination or sequentially, and in particular substantially simultaneously or in whole or in part sequentially. When these steps are performed in whole or in part sequentially, at least one composition (or a first composition and / or a second composition) may be applied together or separately, and each composition may be applied before or after the application of the alternating electric field.

[0056] The methods disclosed herein can be used to treat any type of cancer cell / cancer / tumor that responds to treatment using alternating electric fields (e.g., TTFields), immune checkpoint inhibitors, and / or activators of MHC class I molecule expression. Non-limiting examples of cancer cells / cancer / tumors that can be treated according to this disclosure include hepatocellular carcinoma / hepatocellular carcinoma cells, glioblastoma / glioblastoma cells, pleural mesothelioma / pleural mesothelioma cells, differentiated thyroid carcinoma / differentiated thyroid carcinoma cells, advanced renal cell carcinoma / advanced renal cell carcinoma cells, ovarian cancer / ovarian carcinoma cells, pancreatic cancer / pancreatic carcinoma cells, lung cancer / lung cancer cells, breast cancer / breast cancer cells, etc., and any combination thereof.

[0057] In a specific (but not limiting) implementation, the cancer may be a solid tumor.

[0058] According to the method of this disclosure, an alternating electric field can be generated using any type of conductive or non-conductive electrode and / or transducer array known in the art or otherwise contemplated herein for the purpose of generating an alternating electric field. According to this disclosure, non-limiting examples of electrode and transducer arrays that can be used to generate alternating electric fields include those that are part of an alternating electric field generating system (e.g., a TTField system), such as, but not limited to, those described below: U.S. Patent Nos. 7,016,725, 7,089,054, 7,333,852, 7,565,205, 8,244,345, 8,715,203, 8,764,675, 10,188,851, and 10,441,776; and U.S. Patent Application Nos. US 2018 / 0160933, US 2019 / 0117956, US 2019 / 0307781; and US 2019 / 0308016.

[0059] According to this disclosure, alternating electric fields can be generated at any frequency. For example (but not as a limitation), the alternating electric field may have frequencies of approximately 50 kHz, approximately 60 kHz, approximately 70 kHz, approximately 75 kHz, approximately 80 kHz, approximately 90 kHz, approximately 100 kHz, approximately 105 kHz, approximately 110 kHz, approximately 115 kHz, approximately 120 kHz, approximately 125 kHz, approximately 130 kHz, approximately 135 kHz, approximately 140 kHz, approximately 145 kHz, approximately 150 kHz, approximately 155 kHz, approximately 160 kHz, approximately 165 kHz, approximately 170 kHz, approximately 175 kHz, approximately 180 kHz, approximately 185 kHz, approximately 190 kHz, approximately 195 kHz, approximately 200 kHz, approximately 225 kHz, approximately 250 kHz, approximately 275 kHz, approximately 300 kHz, approximately 325 kHz, approximately 350 kHz, approximately 375 kHz, approximately 400 kHz, approximately 425 kHz, approximately 450 kHz, approximately 475 kHz, approximately 50 kHz, etc. The frequencies of 0 kHz, approximately 550 kHz, approximately 600 kHz, approximately 650 kHz, approximately 700 kHz, approximately 750 kHz, approximately 800 kHz, approximately 850 kHz, approximately 900 kHz, approximately 950 kHz, approximately 1 MHz, approximately 2 MHz, approximately 3 MHz, approximately 4 MHz, approximately 5 MHz, approximately 6 MHz, approximately 7 MHz, approximately 8 MHz, approximately 9 MHz, approximately 10 MHz, etc., and the ranges formed by any of the above values ​​(e.g., the range of approximately 50 kHz to approximately 10 MHz, the range of approximately 50 kHz to approximately 1 MHz, the range of approximately 50 kHz to approximately 500 kHz, the range of approximately 100 kHz to approximately 500 kHz, the range of approximately 150 kHz to approximately 300 kHz, etc.), and the range of two integers that combine two values ​​falling between the above referenced values ​​(e.g., the range of approximately 122 kHz to approximately 313 kHz, the range of approximately 78 kHz to approximately 298 kHz, etc.).

[0060] In certain specific (but not limiting) embodiments, an alternating electric field may be applied at two or more different frequencies. When two or more frequencies are present, each frequency is selected from any of the values ​​above, or a range formed by any of the values ​​above, or a range of two integers falling between two of the values ​​mentioned above.

[0061] In certain specific (but not limiting) implementations, the following frequencies may be used for specific cancers: GBM, approximately 200 kHz; NSCLC, approximately 150 kHz; breast cancer, approximately 200 kHz; pancreatic cancer, approximately 150 kHz; brain metastases from NSCLC, approximately 150 kHz; liver cancer, approximately 150 kHz, etc.

[0062] The alternating electric field can have any electric field strength in the target area / subject / cancer cells, as long as the alternating electric field can function according to this disclosure. For example (but not as a limitation), the alternating electric field can have at least about 1 V / cm, about 1.5 V / cm, about 2 V / cm, about 2.1 V / cm, about 2.2 V / cm, about 2.3 V / cm, about 2.4 V / cm, about 2.5 V / cm, about 2.6 V / cm, about 2.7 V / cm, about 2.8 V / cm, about 2.9 V / cm, about 3 V / cm, or about 3.5 V / cm in the target area / subject / cancer cells. cm, approximately 4V / cm, approximately 4.5V / cm, approximately 5V / cm, approximately 5.5V / cm, approximately 6V / cm, approximately 6.5V / cm, approximately 7V / cm, approximately 7.5V / cm, approximately 8V / cm, approximately 9V / cm, approximately 9.5V / cm, approximately 10V / cm, approximately 10.5V / cm, approximately 11V / cm, approximately 11.5V / cm, approximately 12V / cm, approximately 12.5V / cm, approximately 13V / cm Electric field strengths such as m, approximately 13.5 V / cm, approximately 14 V / cm, approximately 14.5 V / cm, approximately 15 V / cm, approximately 15.5 V / cm, approximately 16 V / cm, approximately 16.5 V / cm, approximately 17 V / cm, approximately 17.5 V / cm, approximately 18 V / cm, approximately 18.5 V / cm, approximately 19 V / cm, approximately 19.5 V / cm, and approximately 20 V / cm, and ranges formed by any of the above values ​​(e.g., a range from approximately 1 V / cm to approximately 20 V / cm, a range from approximately 1 V / cm to approximately 10 V / cm, a range from approximately 1 V / cm to approximately 4 V / cm, etc.), and ranges of two integers that combine to fall between two values ​​in the above references (e.g., a range from approximately 1.1 V / cm to approximately 18.6 V / cm, a range from approximately 1.2 V / cm to approximately 9.8 V / cm, a range from approximately 1.3 V / cm to approximately 4.7 V / cm, etc.). Typically, it is desirable to use the highest possible electric field strength without causing overheating, where the electric field strength is usually limited by temperature measurements.

[0063] In some cases, the electric field in at least a portion of the target region / subject / cancer cell is induced by an applied voltage determined through computer simulation of the target region / subject / cancer cell. In some cases, the electric field in at least a portion of the target region / subject / cancer cell is induced by an applied voltage of at least 50 V RMS (root mean square) or at least 50 V p2p (peak-to-peak), and optionally, the applied voltage is at least 100 V RMS or at least 100 V p2p. In some embodiments, an applied voltage of at least 50 V induces an electric field with a field strength of at least 1 V / cm (e.g., at least 5 V / cm) in at least a portion of the target region / subject / cancer cell.

[0064] An alternating electric field can be applied in a single direction between a pair of arrays, or it can alternate in two or more directions / channels (e.g., front-to-back and left-to-right) between two or more pairs of arrays. For example, certain TTFields devices (such as, but not limited to, the OPTUNE® system (Novocure Limited, St. Helier, Jersey)) operate in both directions to increase the chance that dividing cells will align with the electric field, allowing the field to have the desired anti-mitotic effect. However, it should be understood that the scope of this disclosure also includes applying an alternating electric field in a single direction. As used herein, the term "alternating electric field" will be understood to include application in a single direction / channel as well as application in two or more directions / channels; furthermore, as used herein, the term "alternating electric field" will be understood to include both the application of a single alternating electric field and the continuous application of multiple alternating electric fields over a time duration.

[0065] An alternating electric field can be applied for any continuous or cumulative period of time sufficient to reduce the viability of cancer cells and / or decrease the size of the tumor (and / or prevent the tumor from increasing in size). This period of application of the alternating electric field includes both continuous and cumulative periods. That is, this period of application includes a single treatment session (i.e., continuous application) and multiple treatment sessions with slight rest between sessions (i.e., continuous application within a cumulative period). For example, subjects may be allowed to rest during treatment with the alternating electric field device, and it is anticipated that the device will only be positioned on the body and operated for at least approximately 50%, at least approximately 60%, at least approximately 70%, or at least approximately 80% of the total treatment period (e.g., over a day, a week, two weeks, a month, two months, three months, four months, five months, etc.).

[0066] For example, but not as a limitation, the alternating electric field may be applied for a continuous or cumulative period of at least approximately 1 hour, approximately 2 hours, approximately 3 hours, approximately 4 hours, approximately 5 hours, approximately 6 hours, approximately 7 hours, approximately 8 hours, approximately 9 hours, approximately 10 hours, approximately 11 hours, approximately 12 hours, approximately 15 hours, approximately 18 hours, approximately 21 hours, approximately 24 hours, approximately 27 hours, approximately 30 hours, approximately 33 hours, approximately 36 hours, approximately 39 hours, approximately 42 hours, approximately 45 hours, approximately 48 hours, approximately 51 hours, approximately 54 hours, approximately 57 hours, approximately 60 hours, approximately 63 hours, approximately 66 hours, approximately 69 hours, approximately 72 hours, approximately 75 hours, or approximately 78 hours. The ranges are approximately 81 hours, 84 hours, 87 hours, 90 hours, 93 hours, 96 hours, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 21 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, etc., as well as ranges formed by any of the above values ​​(e.g., a range from approximately 1 hour to approximately 6 months, a range from approximately 1 hour to approximately 7 days, a range from approximately 24 hours to approximately 72 hours, etc.), and ranges of two integers that combine to fall between two values ​​in the above references (e.g., a range from approximately 14 hours to approximately 68 hours, etc.).

[0067] In one particular (but not limiting) embodiment, the alternating electric field is applied for at least about 24 hours within a continuous 48-hour period. In another particular (but not limiting) embodiment, the alternating electric field is applied for at least about 24 hours, wherein the device is positioned on the body and operates for at least about 80% of that period.

[0068] The total duration of applying the alternating electric field can be continuous or intermittent. That is, when the duration of applying the alternating electric field is short (e.g., but not limited to, less than about 12 or 24 hours), the alternating electric field can be applied continuously within that duration. However, when the duration of applying the alternating electric field is long (e.g., but not limited to, a period of about 24 hours or longer), the treatment period can include one or more interruptions during the application period, which separate two or more application portions, whereby the application portions and interruptions combine to form the total application period. When an interruption is present, the alternating electric field is applied for at least about 50%, about 60%, about 70%, about 80%, or about 90% or more of the treatment time, so the interruption typically accounts for only about 10%, about 20%, about 30%, about 40%, about 50%, or less of the treatment time. In one particular (but not limiting) embodiment, when an interruption is present, the interruption typically accounts for about 20% or less of the treatment time, thereby allowing the alternating electric field to be applied for at least about 80% or more of the treatment time. For example, but not as a limitation, the alternating electric field should be applied for at least approximately 19 hours within each 24-hour period. Furthermore, the longer the alternating electric field is applied, the greater the efficiency.

[0069] Suitable types of immune checkpoint inhibitors that may be used according to this disclosure include, but are not limited to, PD-1 inhibitors, PDL-1 inhibitors, CTLA-4 inhibitors, TIM3 inhibitors, TIGIT inhibitors, LAG3 inhibitors, and agonist antibodies against receptors such as, but not limited to, 4-1BB and OX40. Specific (but not limiting) examples of anti-PD-1 IgG4 that may be used according to this disclosure include nivolumab, pembrolizumab, cimipril, spartazumab, tislelizumab, etc. Specific (but not limiting) examples of anti-PD-L1 IgG1 that may be used according to this disclosure include atezolizumab, durvalumab, avelumab, BGB-A333, etc. Specific (but not limiting) examples of anti-CTLA4 compounds that may be used according to this disclosure include ipilimumab, trimemumab, etc. A specific (but not limiting) example of anti-TIM3 IgG4 that may be used according to this disclosure is sabatolimab, etc. Specific (but not limiting) examples of anti-TIGIT IgG1 compounds that may be used according to this disclosure include vimbrolizumab, tirelinumab, etc. Specific (but not limiting) examples of anti-LAG3 compounds that may be used according to this disclosure include piracetamab, ellarimumab, etc., and ecteramod α (soluble LAG-3 protein).

[0070] Any composition known in the art or otherwise contemplated herein as an MHC class I activator for enhancing the expression of MHC class I molecules may be used according to this disclosure, provided that the composition functions as described herein. Suitable MHC class I activators that may be used according to this disclosure include PI3K inhibitors, autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, topotecan, interferon-γ, BRAF inhibitors, MEK inhibitors, vemurafenib, cobimetinib, and any combination thereof.

[0071] Non-limiting examples of PI3K inhibitors that may be used under this disclosure include apelelixib, datolixib, PI-103, picolidexib, bupalixib, cupanixinexib, duvelixib, tacelilixib, piralilixib, votacitabine, nanoripalasib, erblisse, GDC-0326, SF2523, celabelixib, icolidexib, tenalixib, celabelixib, and lenolixib. Akalisci, Passalisci, ME401, Inawalicis, HS-173, PI-3065, GNE-317, CZC24832, GSK2636771, PF-4989216, AZD8186, AMG319, A66, AS-252424, AS-604850, CAY1505, NU75-441, CH51 32799, GDC-0941, TG100-115, TG100713, CH5132799, PX-866, LY294002, XL147, ZSTK 474, BKM-120, BAY80-6946, AZD8835, WX-037, KA2237, CAL-120, INCB050465, INK-11 17. TGR-1202, RP6530, GDC-0032, BYL719, IPI-145, CAL-101, PIK-90, PIK-294, IC-87114, AS-605240, AZD6482, AMG511, ADZ6482, MLN1117, 3-hydroxy-anaminobenzoic acid, psyllium arvense, citronellol, bufotoxin, etc.

[0072] Non-limiting examples of autophagy inhibitors that may be used according to this disclosure include chloroquine, hydroxychloroquine, cyclohexylimide, bafloxacin A1, Lys05, 3-methyladenine (3-MA), LY294002, Wortmannin, leucosterol peptide, E64d, pepsin A, Spautin 1, MRT 67307, MRT 68921, etc.

[0073] Non-limiting examples of HDAC inhibitors that may be used according to this disclosure include: aspirin, entenostat, MS-275, sodium butyrate (NaB, sodium butyrate), succinyl aniline isohydroxamic acid (SAHA; vorinostat), trachostatin A (TSA), valproic acid, perbisbutazone, belistat, epoxetine, danostatine, quinolostatine, moxistat, sodium valproate (NSC93819, sodium valproate), CUDC-101, droxilstat, MC1568, prasinol, divalproate, genostat, M344, romidesin, tadenafil, and skritide (GCK). 1026), Renostat (RAS2410), RGFP966, RG2833, TMP269, Santa Cruz A (CAY10683), Taquimod (ABR-215050), LMK-235, CAY10603, Domatinositol, BG45, BRD72954, TMP196, Tuxexistat, AR-42, Sitathasitol (ACY-241), GSK3117391, Biphenyl-4-sulfonyl chloride, UF010, Cork oxime acid, NKL22, TC-H 106, SR-4370, SIS17, BRD3308, CXD101, etc.

[0074] A non-limiting example of a BRAF inhibitor that may be used under this disclosure is vemurafenib.

[0075] A non-limiting example of a MEK inhibitor that may be used under this disclosure is cobimetinib.

[0076] The compositions disclosed herein (i.e., containing immune checkpoint inhibitors and / or MHC class I activators) may be provided in any formulation form known in the art or otherwise contemplated herein. In certain specific (but not limiting) embodiments, the composition comprises one or more pharmaceutically acceptable carriers (and therefore, the composition may also be referred to as a "pharmaceutical composition"). Non-limiting examples of suitable pharmaceutically acceptable carriers include: water; saline; dextran solution; fructose or mannitol; calcium carbonate; cellulose; ethanol; oils of animal, plant, or synthetic origin; carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; detergents; liposome carriers; nanocarriers; scaffold materials for achieving sustained drug release (such as, but not limited to, hydrogels); buffer solutions such as sodium chloride, saline, phosphate-buffered saline, and / or other physiologically acceptable and / or safe substances; diluents; excipients such as polyethylene glycol (PEG); or any combination thereof. Suitable pharmaceutically acceptable carriers for pharmaceutical formulations are described, for example, in Remington: Science and Practice of Pharmacy, 23rd edition (2020).

[0077] In certain specific (but not limiting) embodiments, the compositions of this disclosure (i.e., containing immune checkpoint inhibitors and / or MHC class I activators) may also contain one or more additional active agents. A variety of active agents that can be used concurrently with alternating electric fields, immune checkpoint inhibitors, and / or MHC class I molecular expression activators are known in the art, and some combination therapies have been FDA approved or are in clinical trials.

[0078] Furthermore, any composition disclosed herein may contain other pharmaceutical agents that allow for administration via a specific route of administration. For example, but not as a limitation, the composition may be formulated for administration via oral, topical, transdermal, parenteral, subcutaneous, intranasal, mucosal, intramuscular, intraperitoneal, intravitreal, intratumoral, and / or intravenous routes. Depending on the route of administration, the composition may contain one or more additional components in addition to the active agent (e.g., immune checkpoint inhibitors and / or MHC class I molecule expression activators and / or adjunctive therapeutic agents). Examples of possible adjunctive compounds include, but are not limited to, fillers, salts, buffers, preservatives, stabilizers, solubilizers, wetting agents, emulsifiers, dispersants, gels, binders, and other materials well known in the art.

[0079] In one particular (but not limiting) embodiment, any of the compositions disclosed herein (i.e., containing immune checkpoint inhibitors and / or MHC class I activators) is administered to a subject by injection or implantation. For example (but not as a limitation), in some cases it may be desirable to administer the composition at a local / regional level to ensure that the composition targets a specific location in the subject's body and inhibits non-specific interactions in other parts of the body; in other cases, more systematic administration may be required.

[0080] Any composition of the disclosed compositions (i.e., containing an immune checkpoint inhibitor and / or an MHC class I activator) may be administered before or after the application of an alternating electric field. In certain specific (but not limiting) embodiments, at least one composition may be administered before the application of an alternating electric field. In certain specific (but not limiting) embodiments, at least one composition may be administered after the application of an alternating electric field. In particular (but not as a limitation), the composition may be administered during the application of an alternating electric field (e.g., before the end of that period of application of the alternating electric field) and / or after the end of the application of the alternating electric field. In certain specific (but not limiting) embodiments, at least one composition may be administered before the application of an alternating electric field, and / or at least one composition may be administered after the application of an alternating electric field (and before or after the end of the application of the alternating electric field).

[0081] For example (but not as a limitation), any composition of the disclosed compositions (i.e., containing immune checkpoint inhibitors and / or MHC class I activators) may be administered before the application of the alternating electric field begins for the following time periods: at least about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 3 hours, about 6 hours, about 9 hours, about 12 hours, about 15 hours, about 18 hours, about 21 hours, about 24 hours, about 27 hours, about 30 hours, about 33 hours, about 36 hours, about 39 hours, about 42 hours, about 45 hours, about 48 hours, about 51 hours, or about 54 hours. The values ​​are approximately 57 hours, 60 hours, 63 hours, 66 hours, 69 hours, 72 hours, 75 hours, 78 hours, 81 hours, 84 hours, 87 hours, 90 hours, 93 hours, 96 hours, 5 days, 6 days, 7 days, etc., and ranges formed by any of the above values ​​(e.g., a range from approximately 24 hours to approximately 96 hours, etc.), and ranges of two integers that combine two values ​​falling between the above-referenced values ​​(e.g., a range from approximately 14 hours to approximately 94 hours, etc.). In one particular (but not limiting) embodiment, at least one composition that enhances the expression of MHC class I molecules is applied at least approximately 24 hours before the application of the alternating electric field begins.

[0082] In other non-limiting examples, any composition of the present disclosure (i.e., containing an immune checkpoint inhibitor and / or an MHC class I activator) may be administered after the application of an alternating electric field has commenced for the following time periods: at least about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 3 hours, about 6 hours, about 9 hours, about 12 hours, about 15 hours, about 18 hours, about 21 hours, about 24 hours, about 27 hours, about 30 hours, about 33 hours, about 36 hours, about 39 hours, about 42 hours, about 45 hours, about 48 hours, about 51 hours, about 54 hours, or about 54 hours. The values ​​are approximately 57 hours, 60 hours, 63 hours, 66 hours, 69 hours, 72 hours, 75 hours, 78 hours, 81 hours, 84 hours, 87 hours, 90 hours, 93 hours, 96 hours, 5 days, 6 days, 7 days, etc., and ranges formed by any of the above values ​​(e.g., a range from approximately 24 hours to approximately 96 hours, etc.), and ranges of two integers falling between two of the above values ​​(e.g., a range from approximately 14 hours to approximately 94 hours, etc.). In one particular (but not limiting) embodiment, at least one composition for increasing the expression of MHC class I molecules is applied at least approximately 24 hours after the application of the alternating electric field.

[0083] In other non-limiting examples, any composition of the present disclosure (i.e., containing immune checkpoint inhibitors and / or MHC) (Class I activator) may be applied after the end of the period of application of the alternating electric field, wherein one or more of the composition are applied during the following time periods after the end of the period: about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 3 hours, about 6 hours, about 9 hours, about 12 hours, about 15 hours, about 18 hours, about 21 hours, about 24 hours, about 27 hours, about 30 hours, about 33 hours, about 36 hours, about 39 hours, about 42 hours, about 45 hours, about 48 hours, about 51 hours, about 54 hours, about 57 hours, about 60 hours, about 63 hours, about 66 hours, about 69 hours, about 72 hours, about 75 hours, about 78 hours, about 81 hours, about 84 hours, about 87 hours, about 90 hours, about 93 hours, about 96 hours, about 5 days, about 6 days, about 7 days, etc.

[0084] In one particular (but not limiting) embodiment, one or more of the compositions disclosed herein (i.e., containing immune checkpoint inhibitors and / or MHC class I activators) are administered within approximately 96 hours after the end of the period.

[0085] The compositions disclosed herein (i.e., containing immune checkpoint inhibitors and / or MHC class I activators) can be administered to cancer cells / subjects at any concentration of the active agent that provides a therapeutically effective concentration. In some non-limiting embodiments, applying an alternating electric field reduces the amount of active agent required to make the treatment effective compared to a normal therapeutically effective amount of the active agent administered in the absence of an alternating electric field. For example, but not as a limitation, the therapeutically effective concentration of the composition may be reduced by at least about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or more relative to doses of compositions known to be therapeutically effective without the application of an alternating electric field. In one particular (but non-limiting) embodiment, the therapeutically effective concentration of the composition is reduced by at least about 50% compared to doses of compositions known to be therapeutically effective in the absence of an alternating electric field.

[0086] The therapeutically effective concentration of each active agent (immune checkpoint inhibitor or MHC class I activator) used according to this disclosure may be, for example (but not limited to): about 1 nM, about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 90 nM, about 100 nM, about 125 nM, about 150 nM, about 175 nM, about 200 nM, about 250 nM, about 300 nM, about 350 nM, about 400 nM, about 450 nM, about 500 nM, about 550 nM, about 600 nM, about 650 nM, about 700 nM, about 750 nM, about 800 nM, about 850 nM, about 900 nM, about 950 nM, about 1 mM, about 2 mM, about 3mM, approximately 4mM, approximately 5mM, approximately 6mM, approximately 7mM, approximately 8mM, approximately 9mM, approximately 10mM, approximately 11mM, approximately 12mM, approximately 13mM, approximately 14mM, approximately 15mM, approximately 16mM, approximately 17mM, approximately 18mM, approximately 19mM, approximately 20mM, approximately 25mM, approximately 30mM, approximately 35mM, approximately 40mM, approximately 45mM, approximately 50mM, etc., and ranges formed by any of the above values ​​(e.g., a range from approximately 12.5nM to approximately 100nM, a range from approximately 1nM to approximately 20nM, etc.), and ranges of two integers that combine two values ​​falling between the above-referenced values ​​(e.g., a range from approximately 17nM to approximately 83nM, etc.).

[0087] In one particular (but not limiting) embodiment, the therapeutically effective concentration of each active agent is from about 10 nM to about 100 nM.

[0088] In certain (but not limiting) embodiments, the therapeutically effective concentration of each active agent used according to this disclosure may be, for example (but not as a limitation): about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, etc. g / kg, approximately 19 mg / kg, approximately 20 mg / kg, approximately 21 mg / kg, approximately 22 mg / kg, approximately 23 mg / kg, approximately 24 mg / kg, approximately 25 mg / kg, approximately 26 mg / kg, approximately 27 mg / kg, approximately 28 mg / kg, approximately 29 mg / kg, approximately 30 mg / kg, approximately 31 mg / kg, approximately 32 mg / kg, approximately 33 mg / kg, approximately 34 mg / kg, approximately 35 mg / kg, approximately 36 mg / kg, approximately 37 mg / kg, approximately 38 mg / kg, approximately 39 mg / kg, approximately 40 mg / kg kg, approximately 41 mg / kg, approximately 42 mg / kg, approximately 43 mg / kg, approximately 44 mg / kg, approximately 45 mg / kg, approximately 46 mg / kg, approximately 47 mg / kg, approximately 48 mg / kg, approximately 49 mg / kg, approximately 50 mg / kg, approximately 51 mg / kg, approximately 52 mg / kg, approximately 53 mg / kg, approximately 54 mg / kg, approximately 55 mg / kg, approximately 56 mg / kg, approximately 57 mg / kg, approximately 58 mg / kg, approximately 59 mg / kg, approximately 60 mg / kg, approximately 65 mg / kg, approximately 70 mg / kg Approximately 75 mg / kg, approximately 80 mg / kg, approximately 85 mg / kg, approximately 90 mg / kg, approximately 95 mg / kg, approximately 100 mg / kg, etc., and ranges consisting of any of the above values ​​(e.g., a range of approximately 10 mg / kg to approximately 50 mg / kg, a range of approximately 1 mg / kg to approximately 40 mg / kg, a range of approximately 1 mg / kg to approximately 30 mg / kg, a range of approximately 1 mg / kg to approximately 25 mg / kg, a range of approximately 1 mg / kg to approximately 20 mg / kg, a range of approximately 1 mg / kg to approximately 10 mg / kg, etc.).

[0089] In certain specific (but not limiting) embodiments, the method includes one or more additional steps. For example (but not as a limitation), the method may also include the step of stopping the application of the alternating electric field (e.g., but not limited to, to allow cell / tissue recovery). Furthermore, any of these steps may be repeated once or multiple times.

[0090] In certain specific (but not limiting) embodiments, any composition of the disclosed compositions (comprising immune checkpoint inhibitors and / or MHC class I activators) may be administered via any dosage regimen known in the art. For example, but not as a limitation, each composition may be administered in a single dose or multiple doses over a defined period of treatment. For example (but not as a limitation), one or more compositions may be administered at therapeutically effective concentrations at the following frequencies: approximately every 4 hours, approximately every 8 hours, approximately every 12 hours, approximately once daily, approximately every other day, approximately every three days, approximately once weekly, approximately twice weekly, approximately three times weekly, approximately once every two weeks, approximately once every three weeks, approximately once monthly, etc., and ranges formed by any of the above values ​​(ranges from approximately every 4 hours to approximately every 8 hours, from approximately once weekly to approximately once monthly, etc.).

[0091] Furthermore, when multiple compositions are administered (i.e., immune checkpoint inhibitors and MHC class I activators are present in different compositions), two or more compositions may be administered via the same route (e.g., both via intravenous administration), or two or more compositions may be administered via different routes (e.g., one composition is administered orally and the other via intravenous administration).

[0092] In certain specific (but not limiting) embodiments, the method involves concurrent therapy using additional supplementary compositions. Therefore, the method may include the additional step of administering at least one supplementary composition (other than a composition containing an immune checkpoint inhibitor and / or an MHC class I molecule expression activator) to cancer cells / subjects.

[0093] When present, the application of at least one additional composition may be substantially simultaneous with or wholly or partially sequentially with any composition containing an immune checkpoint inhibitor and / or an MHC class I molecule expression activator, such that the individual compositions may be applied simultaneously or wholly or partially sequentially. Furthermore, when the method includes the application of the additional composition, the optional application steps may be performed in the same manner and time range as described above for the other compositions, before or after the application of the alternating electric field, during the application of the alternating electric field, and / or after the application of the alternating electric field has ended.

[0094] In certain specific (but non-limiting) embodiments, the method may further include the step of administering at least one additional therapy to the cells / subject. Any therapy known in the art or otherwise contemplated herein for use with alternating electric fields (e.g., TTFields), immune checkpoint inhibitors, and / or MHC class I molecule expression activators may be used according to the methods of this disclosure. Non-limiting examples of additional therapies that may be used include radiotherapy (such as, but not limited to, ionizing radiotherapy), photodynamic therapy, transarterial chemoembolization (TACE), or combinations thereof.

[0095] Furthermore, any step in the procedure may be repeated once or multiple times. Each step within the procedure may be repeated multiple times as needed. When repeating the step of applying the alternating electric field, the transducer array may be placed on the subject in a slightly different location from its original placement; repositioning the array in this way can further aid in the treatment of tumors / cancer. Additionally, any step in the procedure of administering any composition / additional therapy may be repeated a different number of times and at different intervals to follow any known and / or generally accepted dosage / treatment regimen of the composition / therapy.

[0096] While the methods described above involve combining alternating electric fields (e.g., TTFields) with immune checkpoint inhibitors and / or MHC class I molecule expression activators for cancer treatment, it should be understood that the scope of this disclosure is not limited to cancer treatment. Rather, this disclosure covers the treatment of any other related diseases, infections, or conditions for which immune checkpoint inhibitor and / or MHC class I molecule expression activator treatment and / or alternating electric field treatment are beneficial.

[0097] Certain non-limiting embodiments of this disclosure relate to any composition disclosed herein or otherwise contemplated that enhances the expression of MHC class I molecules in cancer cells in a subject, and wherein the composition is used in any of the methods disclosed herein or otherwise contemplated.

[0098] Certain non-limiting embodiments of this disclosure relate to kits containing any components of an alternating electric field (e.g., TTFields) generating system disclosed herein or otherwise contemplated (such as, but not limited to, one or more transducer arrays and / or one or more hydrogel compositions, as disclosed in: U.S. Patent Nos. 7,016,725; 7,089,054; 7,333,852; 7,565,205; 8,244,345; 8,715,203; 8,764,675; 10,188,851; and 10,441,776; and U.S. Patent Application Nos. US 2018 / 0160933; US 2019 / 0117956; US 2019 / 0307781; and US 2019 / 0307781; and US 2019 / 0160933; US 2019 / 0117956; ...60956; and US 2019 / 0160933; US 2019 / 0160956; and US 2019 / 0160956; US 2019 / 0160956; and US 2019 / 0160956; US 2019 / 0160956; and US 2019 / 0160956; US 2019 / 0160956; and US 2019 / 0 The kit may optionally further include one or more of the following compositions (such as, but not limited to, one or more optional compositions containing at least one additional active agent): (e.g., 2019 / 0308016), and any composition disclosed herein or otherwise contemplated (containing an immune checkpoint inhibitor and / or an MHC class I molecule expression activator). The kit may optionally further include one or more devices (or one or more components of a device) used in one or more additional therapeutic steps.

[0099] In one particular (but not limiting) embodiment, the kit may further include instructions for implementing any of the methods disclosed herein or otherwise contemplated. For example (but not as a limitation), the kit may include instructions for applying one or more components of an alternating electric field (e.g., TTFields) generating device to a patient's skin, instructions for applying an alternating electric field to a patient, instructions for formulating one or more compositions in the composition, instructions for when and how to apply one or more compositions, and / or instructions for when to activate and deactivate the alternating electric field relative to the application of the composition and / or optional therapeutic steps.

[0100] In addition to the components described in detail above, the kit may further contain other components / reagents for performing any specific method of the particular method described herein or otherwise contemplated. For example (but not as a limitation), the kit may additionally include: (i) components for preparing the skin prior to treatment of the hydrogel composition and / or the transducer array thereon (e.g., razor, cleaning composition, or wipe / towel, etc.); (ii) components for removing the gel / transducer array; (iii) components for cleaning the skin after removing the gel / transducer array; and / or (iv) other components for use with the system (e.g., conductive materials, non-conductive materials, emollient gels or creams, bandages, etc.). The nature of these additional components / reagents will depend on the specific form of treatment, and their identification is entirely within the skill of a person skilled in the art; therefore, further description of them is not considered necessary. Moreover, the components / reagents present in the kit may each be located in a separate container / compartment, or various components / reagents may be combined and located in one or more containers / compartments, depending on the sterility, cross-reactivity, and stability of the components / reagents.

[0101] The kit can be placed in any packaging that allows the components present therein to function according to this disclosure. In some non-limiting embodiments, the kit also includes a sealed package in which the components are placed. In some specific (but non-limiting) embodiments, the sealed package is substantially airtight and / or substantially lighttight.

[0102] Furthermore, the kit may include a set of written instructions explaining how to use one or more components of the kit. Kits of this nature can be used in any of the methods described herein or otherwise conceived.

[0103] In some non-limiting embodiments, the kit has a shelf life of at least about 6 months, such as (but not limited to) at least about 9 months or at least about 12 months.

[0104] Certain non-limiting embodiments of this disclosure relate to systems that include any components of alternating electric field generating systems disclosed herein or otherwise conceived (such as, but not limited to, one or more transducer arrays and / or one or more hydrogel compositions, as disclosed in: U.S. Patent Nos. 7,016,725; 7,089,054; 7,333,852; 7,565,205; 8,244,345; 8,715,203; 8,764,675; 10,188,851; 10,441,776; and U.S. Patent Application Nos. US 2018 / 0160933; US 2019 / 0117956; US 2019 / 0307781; and US 2019 / 0307781; and US 2019 / 0160933; US 2019 / 0117956; and US 2019 / 0307781; and US 2019 / 0160933; US 2019 / 0117956; and US 2019 / 0307781; and US 2019 / 0160933; US 2019 / 0160956; and ... The system may optionally further include one or more of the following compositions: (as disclosed in document number 2019 / 0308016), and at least one of the following compositions (comprising immune checkpoint inhibitors and / or MHC class I molecule expression activators). The system may optionally further include one or more of the following optional compositions: (as disclosed in document number 2019 / 0308016). The system may optionally further include one or more devices (or one or more components of a device) used in one or more additional therapeutic steps.

[0105] Example

[0106] Examples are provided below. However, this disclosure should be understood to be limited in its application to the specific experiments, results, and laboratory procedures disclosed below. Rather, the examples are provided only as one of various implementations and are exemplary, not exhaustive.

[0107] Example 1

[0108] In this embodiment, the effect of TTFields on the expression of MHC class I molecules in cancer cell lines was investigated. Contrary to the prior art described by Silgner et al. (2018) cited in the background section above, which found that the expression of MHC class I and MHC class II molecules remained unchanged after exposure to TTFields, this embodiment found that the expression of MHC class I molecules in human and mouse pancreatic cancer cell lines was actually reduced after exposure to TTFields. Figure 1 As shown, after 72 hours of TTFields exposure, the expression of MHC class I molecules in the 1A-AsPc1 human pancreatic cancer cell line decreased (left panel), and after 72 hours or 96 hours of TTFields exposure, the expression of MHC class I molecules in the 1B-Panc02 mouse pancreatic cancer cell line decreased (middle and right panels).

[0109] In addition, transcriptomic analysis was conducted to identify class I HLA genes that showed differential expression compared to the control group in various cancer cell lines treated with TTFields. For example... Figure 2 As shown, statistically significant reductions in various class I HLA molecules (including HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, HLA-G, and HLA-H) were detected after TTFields in glioblastoma (GBM) cell lines, malignant pleural mesothelioma (MPM) cell lines, and pancreatic cancer cell lines AsPC1 and BxPC3. Therefore, these data indicate that TTFields reduce the expression of MHC class I molecules in various cancers.

[0110] While not wishing to be bound by any theory, it has been previously shown that TTFields induce ER stress (Silginer et al., Cell Death and Disease (2017), Vol. 8: e2753). Furthermore, it has been previously shown that ER stress downregulates class I HLA molecules (Ulianich et al., (2011), Acta Biochemica et Biophysica Acta, 1812: 431-438). Therefore, TTFields exposure-induced ER stress may be a possible mechanism for the downregulation of class I HLA molecules following TTFields exposure.

[0111] Example 2

[0112] In this embodiment, the combined effects of TTFields and MHC class I molecular expression activators (such as, but not limited to, chloroquine and aperidilxi) were investigated.

[0113] exist Figure 3 In this study, the expression of MHC class I molecules in Panc02-luc mouse pancreatic cancer cells (an orthotopic pancreatic cancer model) treated with the following methods: chloroquine (an autophagy inhibitor) alone, TTFields alone, and a combination of TTFields and chloroquine. It was observed that MHC class I molecule expression decreased in response to TTFields application; however, when untreated cells were simultaneously exposed to a combination of TTFields and an autophagy inhibitor, MHC class I molecule expression was significantly higher than in untreated cells.

[0114] exist Figure 4In this study, MHC class I molecule expression was evaluated in A2780 cells (an ovarian cancer cell line) treated with the following methods: apelexicillin alone (a PI3K inhibitor), TTFields alone, and a combination of TTFields and apelexicillin. It was observed that MHC class I molecule expression decreased in response to TTFields application; however, MHC class I molecule expression in cells exposed to the combination of TTFields and a PI3K inhibitor was not only higher than that in the untreated control group, but also higher than that in the single-treatment groups.

[0115] Example 3

[0116] This embodiment relates to the use of TTFields in combination with a composition containing at least one immune checkpoint inhibitor and at least one MHC class I molecule expression activator for cancer treatment purposes. Concomitant therapy with TTFields and the composition provides a synergistic effect compared to monotherapy.

[0117] Human subjects were treated with TTFields by applying the OPTUNE® device (Novocure Limited, St. Helier, Jersey) to the skin of the subjects, in which a pair of arrays were placed to the left and right (LR) and / or front and back (AP) of the tumor. Each subject was then treated with TTFields for an extended period at 150–200 kHz; the device was worn for at least approximately 80% of the time, with brief breaks and slight adjustments to the array placement between treatments to allow for cellular and skin recovery.

[0118] Two weeks after the start of TTFields administration, human subjects were orally or injected with at least one immune checkpoint inhibitor and at least one MHC class I molecular expression activator at approximately 1-10 mg / kg, approximately 1-3 times per week, for 6-8 weeks.

[0119] The efficacy of the simultaneous therapy on the tumor was evaluated after treatment with the simultaneous therapy.

[0120] Non-limiting exemplary embodiments of the present invention

[0121] Exemplary Implementation 1: A method for reducing the viability of cancer cells, the method comprising the steps of: (1) applying an alternating electric field to the cancer cells for a period of time; and (2) administering at least one composition to the cancer cells, wherein the at least one composition enhances the expression of MHC class I molecules in the cancer cells.

[0122] Exemplary Implementation Scheme 1A: The method according to Exemplary Implementation Scheme 1, wherein the method is an in vitro method.

[0123] Exemplary Implementation Scheme 1B: The method according to Exemplary Implementation Scheme 1, wherein the method is an in vivo method.

[0124] Exemplary Implementation Scheme 2: A method for treating cancer in a subject, the method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; and (2) administering to the subject at least one composition, wherein the at least one composition enhances the expression of MHC class I molecules in cancer cells.

[0125] Exemplary Implementation 3: A method for reducing the volume of a tumor and / or preventing the tumor from increasing in volume, wherein the tumor is present in the body of a living subject and contains multiple cancer cells, the method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; and (2) administering to the subject at least one composition, wherein the at least one composition enhances the expression of MHC class I molecules in the cancer cells.

[0126] Exemplary Implementation 4: A method comprising the steps of: (1) applying an alternating electric field to a target region of a subject for a period of time; and (2) administering at least one composition to the subject, wherein the at least one composition enhances the expression of MHC class I molecules in the cancer cells; and wherein administering the alternating electric field enhances the efficacy of the at least one composition against the cancer cells in the subject compared to administering the at least one composition to the subject without applying an alternating electric field.

[0127] Exemplary Embodiment 5: The method according to any one of Exemplary Embodiments 1 to 4, wherein the at least one composition comprises at least one immune checkpoint inhibitor and at least one compound that enhances the expression of MHC class I molecules in the cancer cells.

[0128] Exemplary Embodiment 6: A method for reducing the viability of cancer cells, the method comprising the steps of: (1) applying an alternating electric field to the cancer cells for a period of time; (2) administering at least one first composition to the cancer cells, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) administering at least one second composition to the cancer cells, wherein the at least one second composition comprises at least one compound that increases the expression of MHCI class molecules in the cancer cells.

[0129] Exemplary Implementation Scheme 6A: The method according to Exemplary Implementation Scheme 6, wherein the method is an in vitro method.

[0130] Exemplary Implementation Scheme 6B: The method according to Exemplary Implementation Scheme 6, wherein the method is performed in vivo.

[0131] Exemplary Implementation 7: A method for treating cancer in a subject, the method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; (2) administering at least one first composition to the subject, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) administering at least one second composition to the subject, wherein the at least one second composition comprises at least one compound that enhances the expression of MHC class I molecules in cancer cells in the subject.

[0132] Exemplary Implementation 8: A method for reducing the volume of a tumor and / or preventing the tumor from increasing in volume, wherein the tumor is present in the body of a living subject and contains multiple cancer cells, the method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; (2) administering at least one first composition to the subject, wherein the at least one composition comprises at least one immune checkpoint inhibitor; and (3) administering at least one second composition to the subject, wherein the at least one second composition comprises at least one compound that increases the expression of MHC class I molecules in cancer cells in the subject.

[0133] Exemplary Implementation 9: A method comprising the steps of: (1) applying an alternating electric field to a target region of the subject for a period of time; (2) administering at least one first composition to the subject, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) administering at least one second composition to the subject, wherein the at least one second composition enhances the expression of MHC class I molecules in cancer cells in the subject; and wherein administering the alternating electric field enhances the toxicity of the at least one first composition and / or the at least one second composition to cancer cells in the subject compared to administering the at least one first composition and / or the at least one second composition to the subject without the application of an alternating electric field.

[0134] Exemplary Implementation Scheme 10: The method according to any one of Exemplary Implementation Schemes 6 to 9, wherein the two application steps are performed simultaneously, or are performed sequentially, either wholly or partially.

[0135] Exemplary Embodiment 11: The method according to any one of Exemplary Embodiments 1 to 10, wherein at least one of the following is satisfied: the alternating electric field is applied at a frequency in the range of about 50 kHz to about 1 MHz; the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cell / target region of the subject; the alternating electric field is induced by an applied voltage of at least 50 V RMS or at least 50 V p2p; and the time period for which the alternating electric field is applied is at least about 50% of a continuous 24-hour period (i.e., at least about 12 consecutive hours of a 24-hour period).

[0136] Exemplary Embodiment 12: The method according to any one of Exemplary Embodiments 1 to 11, wherein the at least one immune checkpoint inhibitor is selected from the group consisting of: PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, TIM3 inhibitors, TIGIT inhibitors, LAG3 inhibitors, and combinations thereof.

[0137] Exemplary embodiment 12A: The method according to any one of exemplary embodiments 1 to 11, wherein the at least one immune checkpoint inhibitor is selected from the group consisting of: TIM3 inhibitors, TIGIT inhibitors, LAG3 inhibitors, and combinations thereof.

[0138] Exemplary Embodiment 13: The method according to Exemplary Embodiment 12, wherein the at least one immune checkpoint inhibitor comprises at least one PD-1 inhibitor, the at least one PD-1 inhibitor being selected from the group consisting of nivolumab, pembrolizumab, cimipril, spartazumab, tislelizumab, and combinations thereof.

[0139] Exemplary Embodiment 14: The method according to Exemplary Embodiment 12 or 13, wherein the at least one immune checkpoint inhibitor comprises at least one PD-L1 inhibitor, the at least one PD-L1 inhibitor being selected from the group consisting of atezolizumab, durvalumab, avelumab, BGB-A333, and combinations thereof.

[0140] Exemplary Embodiment 15: The method according to any one of Exemplary Embodiments 12 to 14, wherein the at least one immune checkpoint inhibitor comprises at least one CTLA4 inhibitor, the at least one CTLA4 inhibitor being selected from the group consisting of ipilimumab, trimemumab, and combinations thereof.

[0141] Exemplary Embodiment 16: The method according to any one of Exemplary Embodiments 12 to 15, wherein the at least one immune checkpoint inhibitor comprises at least one TIM3 inhibitor, the at least one TIM3 inhibitor comprising sabatolimab.

[0142] Exemplary Embodiment 17: The method according to any one of Exemplary Embodiments 12 to 16, wherein the at least one immune checkpoint inhibitor comprises at least one TIGIT inhibitor, the at least one TIGIT inhibitor being selected from the group consisting of: vimbrolizumab, tirelinumab, and combinations thereof.

[0143] Exemplary Embodiment 18: The method according to any one of Exemplary Embodiments 12 to 17, wherein the at least one immune checkpoint inhibitor comprises at least one LAG3 inhibitor, the at least one LAG3 inhibitor being selected from the group consisting of: piracetamab, eralalimab, ateiramod α (soluble LAG-3 protein), and combinations thereof.

[0144] Exemplary Embodiment 19: The method according to any one of Exemplary Embodiments 1 to 18, wherein the at least one immune checkpoint inhibitor is selected from the group consisting of: nivolumab, pembrolizumab, cimiprimab, spartazumab, tislelizumab, atezolizumab, durvalumab, avelumab, BGB-A333, ipilimumab, trimemumab, sabatolimab, vimbrolizumab, tisreliumab, piracetamab, piracetamab, piracetamab, piracetamab, piracetamab, piracetamab, piracetamab, piracetamab, piracetamab, piracetamab (soluble LAG-3 protein), and combinations thereof.

[0145] Exemplary Embodiment 19A: The method according to any one of Exemplary Embodiments 1 to 18, wherein the at least one immune checkpoint inhibitor is selected from the group consisting of: cimiprimab, spartazumab, tislelizumab, BGB-A333, trimemumab, sabatolimab, vimbrolizumab, tisrelimumab, piracetamab, piracetamab, ileramod α (soluble LAG-3 protein), and combinations thereof.

[0146] Exemplary Embodiment 20: The method according to any one of Exemplary Embodiments 1 to 19, wherein the compound that enhances the expression of MHC class I molecules in cancer cells is selected from the group consisting of: PI3K inhibitors, autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, MEK inhibitors, topotecan, interferon-γ, and combinations thereof.

[0147] Exemplary Embodiment 20A: The method according to any one of Exemplary Embodiments 1 to 19, wherein the compound that enhances the expression of MHC class I molecules in cancer cells is selected from the group consisting of: autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, interferon-γ, and combinations thereof.

[0148] Exemplary Embodiment 21. According to the method of Exemplary Embodiment 20, wherein the at least one compound that enhances the expression of MHC class I molecules in cancer cells comprises at least one PI3K inhibitor, said at least one PI3K inhibitor being selected from the group consisting of: apeleliximab, datoliximab, PI-103, picolideximab, bupaliximab, cupanixineximab, duveliximab, tazeximab, piraceximab, votacitrax, nanoripalasiximab, erblisse, GDC-0326, SF2523, serabeliximab, iganniximab, Tenalisceb, Celalisceb, Lenorelisceb, Akalisceb, Passalisceb, ME401, Inawalixib, HS-173, PI-3065, GNE-317, CZC24832, GSK2636771, PF-4989216, AZD8186, AMG319, A66, AS-252424, AS-604850, CAY1505, NU75 -441, CH5132799, GDC-0941, TG100-115, TG100713, CH5132799, PX-866, LY294002, XL147 , ZSTK474, BKM-120, BAY80-6946, AZD8835, WX-037, KA2237, CAL-120, INCB050465, INK-1 117, TGR-1202, RP6530, GDC-0032, BYL719, IPI-145, CAL-101, PIK-90, PIK-294, IC-87114, AS-605240, AZD6482, AMG511, ADZ6482, MLN1117, 3-hydroxy-anaminobenzoic acid, psyllium arvense, citronellol, bufotoxin, and combinations thereof.

[0149] Exemplary Embodiment 21A: The method according to Exemplary Embodiment 20, wherein the at least one method increases MHC in cancer cells Class I molecules express compounds including at least one PI3K inhibitor selected from the group consisting of: bupalixib, cupanicexib, duvelixib, tsavorixib, piracexib, nanoripalixib, erblisse, GDC-0326, celabelixib, icognasib, tenalixib, celabelixib, lenilixib, acalilixib, passalixib, inavolixib, HS-173, PI-3065, CZC24832, PF-4989216, A66, AS-252424, AS-604850, CAY1505, NU75-441, TG100713, LY294002, PIK-90, PIK-294, IC-87114, AS-605240, and combinations thereof.

[0150] Exemplary Embodiment 22: The method according to Exemplary Embodiment 20 or 21, wherein the at least one compound that enhances the expression of MHC class I molecules in cancer cells comprises at least one autophagy inhibitor selected from the group consisting of: chloroquine, hydroxychloroquine, cyclohexylimide, bafloxacin A1, Lys05, 3-methyladenine (3-MA), LY294002, Wortmannin, leucosterol peptide, E64d, pepsin A, Spautin 1, MRT 67307, MRT 68921, and combinations thereof.

[0151] Exemplary Embodiment 23: The method according to any one of Exemplary Embodiments 20 to 22, wherein the at least one compound that enhances the expression of MHC class I molecules in cancer cells comprises at least one HDAC inhibitor, said at least one HDAC inhibitor being selected from the group consisting of: aspirin, entenostat, MS-275, sodium butyrate (NaB, sodium butyrate), succinyl aniline isohydroxamic acid (SAHA; vorinostat), trachostatin A (TSA), valproic acid, parbistat, belistat, epoxetine, danostine, quinolone, moxistat, sodium valproate (NSC 93819, sodium valproate), CUDC-101, draloxetine, MC1568, prasinolone, divalproate, ganostat, M344, romidesin, tadenafil, skritide (GCK). 1026), renostat (RAS2410), RGFP966, RG2833, TMP269, Santa Cruz ester A (CAY10683), taquimod (ABR-215050), LMK-235, CAY10603, domatinnostat, BG45, BRD72954, TMP196, tucistat, AR-42, sistat (ACY-241), GSK3117391, biphenyl-4-sulfonyl chloride, UF010, cork oxime acid, NKL22, TC-H 106, SR-4370, SIS17, BRD3308, CXD101, and combinations thereof.

[0152] Exemplary Embodiment 24: The method according to any one of Exemplary Embodiments 20 to 23, wherein the at least one compound that enhances the expression of MHC class I molecules in cancer cells comprises at least one BRAF inhibitor, and the at least one BRAF inhibitor comprises vemurafenib.

[0153] Exemplary Embodiment 25: The method according to any one of Exemplary Embodiments 20 to 24, wherein the at least one compound that enhances the expression of MHC class I molecules in cancer cells comprises at least one MEK inhibitor, wherein the at least one MEK inhibitor comprises cobimetinib.

[0154] Exemplary Embodiment 26: The method according to any one of Exemplary Embodiments 1 to 25, wherein the at least one compound that enhances the expression of MHC class I molecules in cancer cells comprises at least one substance selected from the group consisting of: chloroquine, hydroxychloroquine, apelexicillin, interferon-γ, topotecan, entenotide, trichostatin A (TSA), succinylanilide isohydroxamic acid (SAHA), sodium butyrate, aspirin, valproic acid, MS-275, vemurafenib, cobimethinib, and combinations thereof.

[0155] Exemplary embodiment 27: The method according to any one of exemplary embodiments 1 to 26, wherein the at least one composition (or the first composition and / or the second composition) is applied before the application of the alternating electric field begins.

[0156] Exemplary embodiment 28: The method according to any one of exemplary embodiments 1 to 27, wherein the at least one composition (or the first composition and / or the second composition) is applied after the application of the alternating electric field has begun.

[0157] Exemplary embodiment 29: The method according to exemplary embodiment 28, wherein the at least one composition (or the first composition and / or the second composition) is applied before the end of the time period during which the alternating electric field is applied.

[0158] Exemplary embodiment 30: The method according to exemplary embodiment 28 or 29, wherein the at least one composition (or the first composition and / or the second composition) is applied after the said time period has ended.

[0159] Exemplary embodiment 31: The method according to any one of exemplary embodiments 6 to 30, wherein at least one of the first composition and the second composition is applied before the application of the alternating electric field begins, and the other composition is applied after the application of the alternating electric field begins.

[0160] Exemplary Implementation 32: The method according to any one of Exemplary Implementations 1 to 31, wherein any step in the steps is repeated once or multiple times.

[0161] Exemplary embodiment 33: The method according to any one of exemplary embodiments 1 to 32, wherein the cancer cells / cancer / tumor are in the form of at least one solid tumor.

[0162] Exemplary Embodiment 34: The method according to any one of Exemplary Embodiments 1 to 33, wherein the cancer / cancer cells are selected from the group consisting of: hepatocellular carcinoma / hepatocellular carcinoma cells, glioblastoma / glioblastoma cells, pleural mesothelioma / pleural mesothelioma cells, differentiated thyroid cancer / differentiated thyroid cancer cells, advanced renal cell carcinoma / advanced renal cell carcinoma cells, ovarian cancer / ovarian cancer cells, pancreatic cancer / pancreatic cancer cells, lung cancer / lung cancer cells, breast cancer / breast cancer cells, and combinations thereof.

[0163] Exemplary Embodiment 35: The method according to any one of Exemplary Embodiments 1 to 34, wherein at least a portion of the cancer cells in the subject's body or at least a portion of the cancer is resistant to treatment with immune checkpoint inhibitors alone.

[0164] Exemplary Embodiment 36: The method according to any one of Exemplary Embodiments 1 to 35, wherein at least a portion of the cancer cells in the subject's body or at least a portion of the cancer is resistant to immunotherapy.

[0165] Exemplary Embodiment 37: A composition for enhancing the expression of MHC class I molecules in cancer cells in a subject, used in the method according to any one of Exemplary Embodiment 1, 36.

[0166] Exemplary Embodiment 38: A kit for use in the method according to any one of Exemplary Embodiments 1 to 36, the kit comprising a composition that enhances the expression of MHC class I molecules in cancer cells in a subject; and an electric field generating device configured to apply an alternating electric field to the cancer cells for a period of time.

[0167] Exemplary Embodiment 39: A system for use in the method according to any one of Exemplary Embodiments 1 to 36, the system comprising a first composition comprising at least one immune checkpoint inhibitor; and a second composition that enhances the expression of MHC class I molecules in cancer cells in a subject.

[0168] Exemplary Embodiment 40: A kit for use in the method according to any one of Exemplary Embodiments 1 to 36, the kit comprising: a first composition comprising at least one immune checkpoint inhibitor; a second composition that enhances the expression of MHC class I molecules in cancer cells in a subject; and an electric field generating device configured to apply an alternating electric field to the cancer cells for a period of time.

[0169] While the appended disclosure describes the inventive concept in conjunction with the specific experiments, results, and language set forth below, many alternatives, modifications, and variations will be apparent to those skilled in the art. Therefore, this disclosure is intended to cover all such alternatives, modifications, and variations that fall within the spirit and broad scope of this disclosure.

Claims

1. A system for use in a method of reducing the viability of cancer cells, the system comprising: A first composition, the first composition comprising at least one immune checkpoint inhibitor; and The second composition enhances the expression of MHC class I molecules in cancer cells in a subject, wherein the second composition is selected from the group consisting of autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, interferon-γ, and combinations thereof.

2. A kit for use in a method of reducing the viability of cancer cells, the kit comprising: A first composition, the first composition comprising at least one immune checkpoint inhibitor; A second composition that enhances the expression of MHC class I molecules in cancer cells in a subject, wherein the second composition is selected from the group consisting of: autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, interferon-γ, and combinations thereof; and An electric field generating device configured to apply an alternating electric field to the cancer cells for a period of time.

3. The system of claim 1 or the kit of claim 2, wherein the at least one immune checkpoint inhibitor is selected from the group consisting of: PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, TIM3 inhibitors, TIGIT inhibitors, LAG3 inhibitors, and combinations thereof.

4. A method for reducing the viability of cancer cells, the method comprising the following steps: (1) Apply an alternating electric field to the cancer cells for a period of time; (2) administering at least one first composition to the cancer cells, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) Applying at least one second composition to the cancer cells, wherein the at least one second composition enhances the expression of MHC class I molecules in the cancer cells, and wherein the at least one second composition is selected from the group consisting of autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, interferon-γ, and combinations thereof.

5. The method of claim 4, wherein the method is performed in vitro.

6. A method for treating cancer in a subject, the method comprising the following steps: (1) Apply an alternating electric field to the target area of ​​the subject for a period of time; (2) administering at least one first composition to the subject, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) The subject is given at least one second composition, wherein the at least one second composition increases the expression of MHC class I molecules in cancer cells in the subject, and wherein the at least one second composition is selected from the group consisting of autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, interferon-γ, and combinations thereof.

7. A method comprising the following steps: (1) Apply an alternating electric field to the target area of ​​the subject for a period of time; (2) administering at least one first composition to the subject, wherein the at least one first composition comprises at least one immune checkpoint inhibitor; and (3) Administering at least one second composition to the subject, wherein the at least one second composition enhances the expression of MHC class I molecules in cancer cells in the subject, and wherein the at least one second composition is selected from the group consisting of: autophagy inhibitors, HDAC inhibitors, TRAF3 inhibitors, BRAF inhibitors, interferon-γ, and combinations thereof; and When the at least one first composition and / or the at least one second composition are administered to the subject without the application of an alternating electric field, the application of the alternating electric field increases the toxicity of the at least one first composition and / or the at least one second composition to cancer cells in the subject.

8. The method according to any one of claims 4 to 7, wherein at least one of the following is satisfied: The alternating electric field is applied at a frequency in the range of approximately 50 kHz to approximately 1 MHz; The alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells / subject; and The time period during which the alternating electric field is applied is at least about 50% of a continuous 24-hour period.

9. The method according to any one of claims 4 to 8, wherein the at least one immune checkpoint inhibitor is selected from the group consisting of: PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, TIM3 inhibitors, TIGIT inhibitors, LAG3 inhibitors, and combinations thereof.

10. The method according to any one of claims 4 to 9, wherein steps (1) and (2) are performed in whole or in part sequentially, and wherein the at least one first composition and the at least one second composition are applied after the application of the alternating electric field has begun.

11. The method according to any one of claims 4 to 10, wherein the cancer cells are selected from the group consisting of hepatocellular carcinoma cells, glioblastoma cells, pleural mesothelioma cells, differentiated thyroid cancer cells, advanced renal cell carcinoma cells, ovarian cancer cells, pancreatic cancer cells, lung cancer cells, breast cancer cells, and combinations thereof.

12. The method according to any one of claims 6 to 11, further defined as a method for reducing the volume of a tumor and / or preventing the tumor from increasing in volume, wherein the tumor is present in the body of a living subject and contains multiple cancer cells.

13. The method according to any one of claims 4 to 12, wherein at least a portion of the cancer cells are resistant to treatment with immune checkpoint inhibitors alone.

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