Compositions, systems and methods for treating cancer using tumor therapy electric field and VEGF inhibitor

Abstract

Compositions, systems, and methods for reducing cancer cell viability and treating cancer, as well as preventing an increase in the volume of a tumor present in a living subject are disclosed. The systems and methods comprise applying an alternating field and administering at least one composition comprising at least one small molecule anti-angiogenic agent that specifically interacts with VEGF in combination.

Description

Background Art

[0001] Tumor Treatment Fields (TTFields) are low intensity (e.g., 1-3 V / cm) alternating electric fields in the mid-frequency range (e.g., but not limited to, 100-500 kHz) that target solid tumors by disrupting mitosis. This non-invasive treatment targets solid tumors and is described, for example, in U.S. Patents 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 perpendicular fields within the tumor being treated; the electrode arrays that make up each of these electrode pairs are located on opposite sides of the body part being treated. More specifically, for In the TTFields system, one pair of electrodes is located at the left and right sides of the tumor (LR) and the other pair of electrodes is located at the anterior and posterior sides of the tumor (AP). TTFields is approved for the treatment of glioblastoma multiforme (GBM) and can be used, for example, The TRANSPORT(R) system (Novocure Limited, St. Helier, Jersey) is delivered with a transducer array that is placed on the patient's shaved head.

[0002] Used in Each transducer array in the device that delivers TTFields consists of a set of ceramic disc electrodes that are coupled to the patient's skin (for example, but not limited to, a patient who has shaved his head for treatment of GBM) through a layer of conductive medical gel. The medical gel is intended to deform to match the contours of the body and provide good electrical contact between the array and the skin; in this way, the gel interface bridges the skin and reduces interference. The device is intended to be worn continuously by the patient for 2-4 days, then removed for hygiene care and re-shaving (if necessary), and then reapplied with a new set of arrays. In this way, the medical gel remains in essentially continuous contact with the patient's skin area for 2-4 days at a time. In addition, the array can be moved several centimeters in either direction to allow the skin to heal between one treatment period and the next. Thus, a portion of the skin that was covered by the electrode / gel for 2-4 days can be exposed for 2-4 days while the replacement electrode moves slightly; the device can then be reapplied to the original portion of skin during the next 2-4 days.

[0003] Angiogenesis is the process of forming new blood vessels to increase the supply of nutrients and oxygen to areas of the body that have lost blood supply. Angiogenesis has been shown to be one of the key processes in the hallmarks of cancer. In addition, vascular endothelial growth factor (VEGF) has been shown to be one of the key players in promoting angiogenesis.

[0004] In considering TTFields as a potential improved treatment for osteosarcoma, Oh et al. (Technology in Cancer Research & Treatment (2020) doi: 10.1177 / 1533033820947481) demonstrated that TTFields prevented angiogenesis in human tumor endothelial cells and also downregulated the expression of VEGF and matrix metalloproteinase-2 (MMP2). In addition, Tang et al. (J Int Med Res (2012) 40 (1): 85-94) demonstrated that in mouse melanoma cell lines and mouse tumor models, a decrease in VEGF expression was observed when exposed to moderate alternating electric fields. In addition, Kim et al. (Oncotarget (2016) 7: 65125-65136) demonstrated that TTFields inhibited glioblastoma cell migration, invasion, and angiogenesis, and claimed that TTFields represents a promising anti-invasive and anti-angiogenic therapeutic strategy for GBM patients.

[0005] Despite the availability of tumor treating field-based therapies, glioblastoma multiforme (GBM) remains the most common and aggressive primary malignancy of the central nervous system in adults. The current standard of care for recurrent GBM is bevacizumab ( Genentech, Inc., San Francisco, CA), a humanized monoclonal antibody against vascular endothelial growth factor A (VEGF-A). Emerging preclinical and clinical data suggest that anti-VEGF-A therapy is potentially effective in GBM (Shiyu et al., Biomedicine & Pharmacotherapy (2021) 141: 111810). However, patients inevitably develop resistance to bevacizumab and generally fail to show significantly better overall survival.

[0006] In Ansstas et al. (Case Rep Neurol (2016) 8:1-9), a “pulse dose” approach to bevacizumab administration was combined with TTFields therapy, in which patients with recurrent GBM discontinued bevacizumab therapy and then received TTFields therapy alone, followed by bevacizumab again in a “pulse dose” manner once symptoms and / or evidence of radiographic progression developed. The results of this study support the use of TTFields therapy with pulse doses of bevacizumab in patients with refractory GBM. Fallah et al. (J Clin Oncology (2020) 38(15_suppl):2537), the combination of bevacizumab and TTFields was shown to be safe, feasible, and clinically effective in patients with recurrent GBM.

[0007] Davidi et al. (Journal of Radiation Oncology (2021) 111(3):e47-e48), Gkika et al. (Cancers (Basel) (2022) 14(6):1568), and Davidi et al. (Cancers (Basel) (2022) 14(12):2959) have combined TTFields therapy with sorafenib (a multikinase inhibitor and the leading first-line treatment for advanced HCC) to treat hepatocellular carcinoma (HCC, a highly malignant liver cancer and one of the leading causes of cancer-related death worldwide). In patients with advanced HCC, the TTFields / sorafenib combination showed improved response rates compared to historical controls.

[0008] Jo et al. (Int J Mol Sci (2018) 19(11):3684) investigated the effects of sorafenib on the antitumor and antiangiogenic activities of TTFields in glioblastoma cells and found that the combined treatment inhibited the motility, invasiveness and angiogenesis of tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 Included are representative images of ovaries with tumors in a tropism ovarian cancer mouse model after 10 days of treatment with TTFields at 200 kHz.

[0010] Figure 2 showed that serum circulating VEGF expression was upregulated after 10 days of treatment with TTFields at 200 kHz in an ovarian cancer mouse model.

[0011] Figure 3 showed that in a mouse model of lung cancer, serum circulating VEGF expression was upregulated after 14 days of treatment with TTFields at 150kHz.

[0012] Figure 4 showed that intratumoral CD31 expression was upregulated after 10 days of treatment with TTFields at 200 kHz in an ovarian cancer mouse model.

[0013] Figure 5 Included are micrographs of A2780 cells after treatment with TTFields alone, with 4 μM Lenvatinib, and with a combination of TTFields and 4 μM Lenvatinib.

[0014] Figure 6 Demonstrating the effect of combined treatment with TTFields and 4 μM lenvatinib on cell count and apoptosis in the A2780 cell line. DETAILED DESCRIPTION

[0015] Before explaining at least one embodiment of the inventive concept in detail 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 concept of the present invention is capable of other embodiments or practice or execution in various ways. 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. Moreover, it should be understood that the words and terms used herein are for descriptive purposes and should not be considered restrictive.

[0016] Unless otherwise defined herein, scientific and technical terms used in conjunction with the inventive concepts disclosed herein shall have the meanings commonly understood by those of ordinary skill in the art. In addition, unless the context otherwise requires, singular terms shall include the plural, and plural terms shall include the singular. The aforementioned techniques and procedures are generally performed according to conventional methods well known in the art, and as described in various general and more specific references cited and discussed throughout this specification. The nomenclature used in connection with analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry described herein, as well as laboratory procedures and techniques, are those well known and commonly used in the art. Standard techniques are used for chemical synthesis and chemical analysis.

[0017] All patents, published patent applications, and non-patent publications mentioned in this specification are indicative of the skill levels of those skilled in the art to which the inventive concepts of the present invention pertain. 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 same extent as if each individual patent or publication was specifically and individually indicated to be incorporated by reference.

[0018] All compositions, assemblies, systems, kits and / or methods disclosed herein can be prepared and performed without undue experimentation according to the present disclosure. Although the compositions, assemblies, systems, kits and methods of the inventive concept have been described according to specific embodiments, it will be apparent to those skilled in the art that the steps or step sequences of the compositions and / or methods and methods described herein may be varied without departing from the concept, spirit and scope of the inventive concept. All such similar substitutions and modifications 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.

[0019] As used in accordance with the present invention, unless otherwise indicated, the following terms shall be understood to have the following meanings:

[0020] When used in conjunction with the term "comprising" in the claims and / or in the specification, the use of the term "a" or "an" may mean "one", but it is also consistent with the meaning of "one or more", "at least one", and "one or more than one". Thus, the terms "a", "an", and "the" include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to "a compound" may refer to one or more compounds, two or more compounds, three or more compounds, four or more compounds, or an additional number of compounds. The term "plurality" means "two or more".

[0021] The use of the term "at least one" should be understood to include one and any amount 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 term to which it is connected; in addition, the amount of 100 / 1000 is not considered to be limiting, as higher limits can also produce satisfactory results. In addition, the use of the term "at least one of X, Y, and Z" will be understood to include X alone, Y alone, and Z alone, as well as any combination of X, Y, and Z. The use of ordinal terms (e.g., "first," "second," "third," "fourth," etc.) is only for the purpose of distinguishing two or more items and is not meant to imply any order or sequence or importance of one item relative to another, or any order of addition, for example.

[0022] The term "or" as used in the claims is intended to mean the inclusive "and / or" unless explicitly stated to refer to only alternatives or unless the alternatives are mutually exclusive. For example, the condition "A or B" is satisfied by any of the following: A is true (or present) and B is false (or absent), A is false (or absent) and B is true (or present), and both A and B are true (or present).

[0023] As used herein, any reference to "one embodiment," "an embodiment," "some embodiments," "an example," "for example," or "an example" means that a particular element, feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. For example, the appearances of the phrase "in some embodiments" or "an example" in different places in the specification do not necessarily all refer to the same embodiment. Furthermore, all references to one or more embodiments or examples should be construed as non-limiting to the claims.

[0024] In this application, the term "about" is used to indicate that a value includes the inherent error variation of the composition / device / apparatus, the method used to determine the value, or the variation that exists in the study subjects. For example, and not as a limitation, when the term "about" is used, the specified value can vary by plus or minus 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 from the specified value as such variations are appropriate for performing the disclosed methods and as understood by one of ordinary skill in the art.

[0025] As used in the specification and claims, the words "comprising" (and any forms of including, such as "comprise" and "comprises"), "having" (and any forms of having, such as "have" and "has"), "including" (and any forms of including, such as "includes" and "include"), or "containing" (and any forms of including, such as "contains" and "contain") are inclusive or open-ended, and do not exclude additional, unrecited elements or method steps.

[0026] As used herein, the term "or combinations thereof" refers to all permutations and combinations of the items listed before the term. For example, "A, B, C, or combinations thereof" is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if the order is important in the particular context, BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with the example, explicitly included are combinations that include 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 otherwise apparent from the context.

[0027] As used herein, the term "substantially" means that the subsequently described event or circumstance occurs completely or that the subsequently described event or circumstance occurs to a large extent. For example, when associated with a particular event or circumstance, the term "substantially" means that the subsequently described event or circumstance occurs 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 the two items are very close to each other but not 100% adjacent to each other, or that a portion of one of the two items is not 100% adjacent to the other item but is very close to the other item.

[0028] The term "pharmaceutically acceptable" refers to compounds and compositions that are suitable for administration to humans and / or animals without undue adverse side effects (such as (but not limited to) toxicity, irritation and / or allergic response), which are commensurate with a reasonable benefit / risk ratio.

[0029] The terms "patient" or "subject" as used herein include both human and veterinary subjects. "Mammal" for therapeutic purposes refers to any animal classified as a mammal, including but not limited to humans, domestic and farm animals, non-human primates, and any other animal with mammary tissue.

[0030] The term "treatment" refers to both therapeutic treatment and prophylactic or preventative measures. Individuals in need of treatment include, but are not limited to, individuals who already have a particular condition / disease / infection and individuals at risk of acquiring a particular condition / disease / infection (e.g., individuals in need of prophylactic / preventative measures). The term "treatment" refers to the administration of an agent / element / method to a patient for therapeutic and / or prophylactic / preventative purposes.

[0031] As used herein, the term "therapeutic composition" or "pharmaceutical composition" refers to an agent that can be administered in vivo to produce a therapeutic and / or prophylactic / preventive effect.

[0032] Administration of a therapeutically effective amount or a prophylactically effective amount is intended to provide a therapeutic benefit in the treatment, prevention, and / or control of a disease, disorder, and / or infection. The specific amount that is therapeutically effective can be readily determined by an ordinary physician and can vary according to factors known in the art, such as, but not limited to, the type of disorder / disease / infection, the patient's medical history and age, the stage of the disorder / disease / infection, and the co-administration of other agents.

[0033] The term "effective amount" refers to the amount of a bioactive molecule or its conjugate or derivative, or the amount of a treatment regimen (e.g., an alternating electric field), which is sufficient to show a detectable therapeutic effect without undue adverse side effects (e.g., but not limited to, toxicity, irritation, and allergic reactions), commensurate with a reasonable benefit / risk ratio when used in the manner of the inventive concept. The therapeutic effect may include, for example, but not limited to, prevention, inhibition, or reduction of at least one disorder, disease, and / or infection. The effective amount of the subject will depend on the type of subject, the size and health of the subject, the nature and severity of the disorder / disease / infection to be treated, the method of administration, the duration of treatment, the nature of concurrent therapy (if any), the specific formulations used, etc. Therefore, it is impossible to pre-specify an accurate effective amount. However, the effective amount in a given situation can be determined by a person of ordinary skill in the art using routine experiments based on the information provided herein.

[0034] As used herein, the term "concurrent therapy" is used interchangeably with the terms "combination therapy," "concomitant therapy," and "adjuvant therapy," and is understood to mean treating a patient in need of treatment or administering another drug for a condition / disease / infection in conjunction with the treatment of the present invention. Concurrent therapy can be sequential therapy, in which a patient is first treated with one therapeutic regimen / drug composition and then with another therapeutic regimen / drug composition, or both therapeutic regimens / drug compositions are administered simultaneously.

[0035] The terms "administration" and "administering" used herein are 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 include local and systemic administration. In addition, the compositions of the present invention (and / or methods of administration thereof) can be designed to provide delayed, controlled or sustained release using formulation techniques well known in the art.

[0036] The term "small molecule" as used herein refers to a natural or artificial compound with a well-defined structure that binds to a specific biomacromolecule and acts as an effector, altering the activity or function of the target to which it binds. Small molecules typically have a molecular weight generally ranging from about 1 Da to about 1.5 kDa, and if necessary, their small size allows the molecule to diffuse through the cell membrane and reach the site of action within the cell. Small molecules are typically smaller than nucleic acids, proteins, enzymes, antibodies, polysaccharides, biologics, and other biotherapeutic modalities, and their size is typically greater than 1.5 kDa.

[0037] Turning now to the inventive concepts, combination therapies for cancer are disclosed herein. The combination therapy includes the use of alternating electric fields (e.g., but not limited to TTFields) in combination with at least one small molecule anti-angiogenic agent, which binds to vascular endothelial growth factor (VEGF; e.g., but not limited to VEGF-A, VEGF-B, VEGF-C, or VEGF-D) or VEGF receptors (VEGFR; e.g., but not limited to VEGFR-1, VEGFR-2, or VEGFR-3) and inhibits the interaction between VEGF and VEGFR, thereby inhibiting the VEGF signaling pathway. The combination of alternating electric fields (e.g., TTFields) and small molecule anti-angiogenic agents provides synergistic results in cancer treatment.

[0038] Certain non-limiting embodiments of the present invention relate to methods for reducing the viability of cancer cells. The methods include the following steps: (1) administering at least one composition to cancer cells, wherein the at least one composition includes at least one small molecule anti-angiogenic agent, which specifically and physically interacts with VEGF or VEGFR and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits the signal transduction of VEGFR; and (2) applying an alternating electric field to the cancer cells for a period of time.

[0039] Certain additional non-limiting embodiments of the present invention relate to methods for treating cancer in a subject. The methods include the following steps: (1) administering to the subject at least one composition, wherein the at least one composition includes at least one small molecule anti-angiogenic agent, which specifically and physically interacts with VEGF or VEGFR and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits signal transduction of VEGFR; and (2) applying an alternating electric field to a target area of ​​the subject.

[0040] Certain additional non-limiting embodiments of the present invention relate to a method for reducing the volume of a tumor present in a living subject, wherein the tumor comprises a plurality of cancer cells. The method comprises the following steps: (1) administering to the subject at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, which specifically and physically interacts with VEGF or VEGFR and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits signal transduction of VEGFR; and (2) applying an alternating electric field to a target area of ​​the subject.

[0041] Certain additional non-limiting embodiments of the present invention relate to a method for preventing an increase in the volume of a tumor present in a living subject, wherein the tumor comprises a plurality of cancer cells. The method comprises the following steps: (1) administering to the subject at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, which specifically and physically interacts with VEGF or VEGFR and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits the signal transduction of VEGFR; and (2) applying an alternating electric field to a target area of ​​the subject.

[0042] The small molecule anti-angiogenic agents used according to the present invention are defined as specifically and physically interacting with VEGF or VEGFR and selectively inhibiting the interaction between VEGF and VEGFR and / or inhibiting the signal transduction of VEGFR. "Specific and physical interaction" means that there is an actual physical interaction between the small molecule anti-angiogenic agent and VEGF that inhibits / substantially prevents the interaction between VEGF and VEGFR, or there is an actual physical interaction between the small molecule anti-angiogenic agent and VEGFR that inhibits / substantially prevents the interaction between VEGFR and VEGF. This "specific interaction" occurs between the specific structure of the small molecule anti-angiogenic agent and VEGF or VEGFR, in contrast to a "non-specific interaction" that is independent of the specific structure of the small molecule anti-angiogenic agent and VEGF / VEGFR. "Selective inhibitory interaction" means that the small molecule anti-angiogenic agent preferentially binds to VEGF, such as through IC 50 value, and in a manner that inhibits its ability to interact with its VEGFR, or small molecule anti-angiogenic agents preferentially bind to VEGFR, such as by IC 50 The small molecule anti-angiogenic agent is determined by the value of the small molecule anti-angiogenic agent, and in a manner that inhibits its ability to interact with its VEGF. However, it should be understood that the phrases "specifically and physically interact with VEGF or VEGFR" and "selectively inhibit the interaction between VEGF and VEGFR and / or inhibit the signal transduction of VEGFR" do not mean that the small molecule anti-angiogenic agent only binds to / interacts with VEGF or VEGF; in fact, some small molecule anti-angiogenic agents used according to the present invention are multi-kinase inhibitors, which can also "specifically and physically interact" with other ligands or receptors and "selectively inhibit" the interaction between ligands / receptors.

[0043] In certain specific (but non-limiting) methods, at least one composition has a half maximal inhibitory concentration (IC 50) selectively inhibits the interaction between at least one VEGF / VEGFR ligand-receptor pair, with a half-maximal inhibitory concentration (IC) of, for example but not limited to, less than about 14 nM, about 13 nM, about 12 nM, about 11 nM, about 10 nM, about 9 nM, about 8 nM, about 7 nM, about 6 nM, about 5 nM, about 4 nM, about 3 nM, about 2 nM, about 1 nM, etc. In a specific (but non-limiting) embodiment, at least one composition has a half-maximal inhibitory concentration (IC) of less than about 10 nmol / L (10 nM). 50 ) selectively inhibits the interaction between at least one VEGF / VEGFR ligand-receptor pair.

[0044] Steps (1) and (2) of any method of the present invention may be performed simultaneously, 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 comprising at least one small molecule anti-angiogenic agent may be applied before or after the start of application of the alternating electric field.

[0045] The methods of the present invention can be used to treat any type of cancer cell / cancer / tumor that responds to treatment with alternating electric fields (e.g., TTFields) and / or small molecule anti-angiogenic agents. Non-limiting examples of cancer cells / cancers / tumors that can be treated according to the present invention include hepatocellular carcinoma, glioblastoma, pleural mesothelioma, differentiated thyroid cancer, advanced renal cell carcinoma, ovarian cancer, pancreatic cancer, lung cancer, breast cancer, etc., and any combination thereof.

[0046] According to the methods of the present invention, any type of conductive or non-conductive electrodes and / or transducer arrays known in the art or contemplated herein that can be used to generate an alternating electric field may be used to generate the alternating electric field. Non-limiting examples of electrodes and transducer arrays that can be used to generate alternating electric fields according to the present invention include those used as part of a TTFields system, such as, but not limited to, 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 11,452,863; and in U.S. Patent Application Nos. US2018 / 0001078; US2018 / 0160933; US2019 / 0117956; US2019 / 0307781; and US2019 / 0308016.

[0047] According to the present invention, the alternating electric field can be generated at any frequency. For example (but not by way of limitation), the alternating electric field can have a frequency of about 50kHz, about 75kHz, about 100kHz, about 125kHz, about 150kHz, about 175kHz, about 200kHz, about 225kHz, about 250kHz, about 275kHz, about 300kHz, about 325kHz, about 350kHz, about 375kHz, about 400kHz, about 425kHz, about 450kHz, about 475kHz, about 500kHz, about 550kHz, about 600kHz, about 650kHz, about 700kHz, about 750kHz, about 800kHz, about 850kHz, about 900kHz, about 950kHz, about 10 ... Frequencies of about 1 MHz, about 2 MHz, about 3 MHz, about 4 MHz, about 5 MHz, about 6 MHz, about 7 MHz, about 8 MHz, about 9 MHz, about 10 MHz, and the like, and ranges formed by any of the above values ​​(e.g., a range of about 50 kHz to about 10 MHz, a range of about 50 kHz to about 1 MHz, a range of about 50 kHz to about 500 kHz, a range of about 100 kHz to about 500 kHz, a range from about 150 kHz to about 300 kHz, and the like), and ranges combining two integers falling between two of the above values ​​(e.g., a range from about 122 kHz to about 313 kHz, a range from about 78 kHz to about 298 kHz, and the like).

[0048] In certain specific (but non-limiting) embodiments, the alternating electric field may be applied at two or more different frequencies. When there are two or more frequencies, each frequency is selected from any of the above values, or a range formed by any of the above values, or a range of two integers falling between two of the above values.

[0049] The alternating electric field may have any field strength in the subject / cancer cell as long as the alternating electric field is able to function according to the present invention. For example, but not by way of limitation, the alternating electric field may have a value of at least about 1 V / cm, about 1.5 V / cm, about 2 V / cm, about 2.5 V / cm, about 3 V / cm, about 3.5 V / cm, about 4 V / cm, about 4.5 V / cm, about 5 V / cm, about 5.5 V / cm, about 6 V / cm, about 6.5 V / cm, about 7 V / cm, about 7.5 V / cm, about 8 V / cm, about 9 V / cm, about 9.5 V / cm, about 10 V / cm, about 10.5 V / cm, about 11 V / cm, about 11.5 V / cm, about 12 V / cm, about 12.5 V / cm, about 13 V / cm, about 13.5 V / cm, about 14 V / cm, about 14.5 V / cm, about 15 V / cm, about The present invention relates to a field strength of about 15.5 V / cm, about 16 V / cm, about 16.5 V / cm, about 17 V / cm, about 17.5 V / cm, about 18 V / cm, about 18.5 V / cm, about 19 V / cm, about 19.5 V / cm, about 20 V / cm, and a range formed by any of the above values ​​(for example, a range of about 1 V / cm to about 20 V / cm, a range of about 1 V / cm to about 10 V / cm, a range of about 1 V / cm to about 4 V / cm, etc.), as well as a range combining two integers falling between two of the above values ​​(for example, a range of about 1.1 V / cm to about 18.6 V / cm, a range of about 1.2 V / cm to about 9.8 V / cm, a range of about 1.3 V / cm to about 4.7 V / cm, etc.).

[0050] The alternating electric field can be applied in a single direction between a pair of arrays, or can alternate in two directions / channels (e.g., front-to-back and side-to-side) between two pairs of arrays. For example, some TTFields devices (such as, but not limited to, The system (Novocure Limited, St. Helier, Jersey) operates in two directions to increase the chance that dividing cells will align with the electric field so that the electric field can have the desired anti-mitotic effect. However, it should be understood that the scope of the present invention also includes the application of an alternating electric field in a single direction. The term "alternating electric field" as used herein will be understood to include application in a single direction / channel as well as in two directions / channels; furthermore, the term "alternating electric field" as used herein will be understood to include the application of a single alternating electric field as well as the continuous application of multiple alternating electric fields over a period of time.

[0051] The alternating electric field may be applied for any continuous or cumulative time period sufficient to achieve a reduction in cancer cell viability and / or a reduction in tumor volume (and / or prevent an increase in tumor volume). The time period for applying the alternating electric field includes a continuous time period and a cumulative time period. That is, the time period for applying the alternating electric field includes a single session (i.e., continuous application) and multiple sessions with smaller interruptions between sessions (i.e., continuous application of cumulative time periods). For example, the subject is allowed to rest during treatment with the alternating electric field device, and it is expected that the device will only be placed on the body and operated within at least about 60%, at least about 70%, or at least about 80% of the total treatment period (e.g., during a day, a week, two weeks, a month, two months, three months, four months, five months, etc.).

[0052] For example, but not by way of limitation, the alternating electric field can be applied for at least about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 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 76 hours, about 77 hours, about 78 hours, about 79 hours, about 80 hours, about 81 hours, about 82 hours, about 83 hours, about 84 hours, about 85 hours, about 86 hours, about 87 hours, about 88 hours, about 89 hours, about 90 hours, about 91 hours, about 92 hours, about 93 hours, about 94 hours, about 95 hours, about 96 hours, about 97 hours, about 98 hours, about 99 hours, about 100 hours, about 101 hours, about 102 hours, about 103 hours, about 104 hours, about 105 hours, about 106 hours, about 107 hours, about 108 hours, about 109 hours, about 110 hours, about 111 hours, about 12 8 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, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 21 days, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, etc., and ranges formed by any of the above values ​​(for example, a range from about 1 hour to about 6 months, a range from about 24 hours to about 72 hours, etc.), and ranges combining two integers falling between two of the above values ​​(for example, a range from about 14 hours to about 68 hours, etc.).

[0053] In a specific (but non-limiting) embodiment, the alternating electric field is applied for a period of time of at least about 24 hours.

[0054] Any small molecule anti-angiogenic agent that specifically interacts with VEGF or VEGFR and / or specifically inhibits signal transduction of VEGFR, and small molecule anti-angiogenic agents are known in the art or otherwise contemplated herein, as long as the agent is capable of acting as an anti-angiogenic agent that reduces the formation of new vasculature in cancer cells / tumors, and wherein such anti-angiogenic effect occurs through direct interaction of the molecule with VEGF or VEGFR to selectively inhibit the interaction between VEGF and its corresponding VEGFR and thereby inhibit VEGF signaling. Non-limiting examples of small molecule anti-angiogenic agents that can be used in accordance with the present invention include lenvatinib, axitinib, regorafenib, cabozantinib, anlotinib, pazopanib, albendazole, delitinib, motesanib, aflibercept, ponatinib, cediranib, tivozanib, telatinib, apatinib, sematinib, fruquintinib, delitinib, anlotinib, taxifolin, surufatinib, dovitinib, ningetinib, AZD2932, LY2874455, MGCD-265 analogs, ZM 306416, ZM 323881, KRN 633, YF-452, ODM-203, AEE 788, BMS 605541, MAZ51, Ki 8751, SU 5402, SU 5408, SU5205, SU5214, SU 6668, SU14813, XL 092, XL 184, BAW2881, BFH772, A-13958, SKLB1002, WAY-340935, ZD-4190, hVEGF-IN-1, R1530, VEGF-Grab, soluble vascular endothelial growth factor decoy receptor FP3, VEGF-decoy receptor fusion protein, VEGF-decoy receptor and combinations thereof.

[0055] Some specific (but non-limiting) examples of small molecule agents useful in the present invention along with their ICs for VEGFR-1, VEGFR-2 and / or VEGFR-3 are 50 The values ​​are provided in Table 1.

[0056] Table 1: IC values ​​of small molecule anti-angiogenic agents for VEGFR-1, -2 and -3 50 value

[0057]

[0058] In contrast, the previously used sorafenib has an IC of 1.0 for VEGFR-1, VEGFR-2, and VEGFR-3. 50 The values ​​are 21 nM, 90 nM and 16 nM, respectively. At least one small molecule anti-angiogenic agent of the present invention has an IC of about 10 nM or less for one, two or three of VEGFR-1, VEGFR-2 and VEGFR-3.50 Value, and / or at least one small molecule anti-angiogenic agent of the present invention specifically binds (rather than just preferentially binds) only one of VEGF or VEGFR, as described in detail herein. For example, but not limited to, aflibercept is a soluble fusion protein of some human extracellular domains of VEGFR-1 and VEGFR-2 with the Fc portion of human immunoglobulin (Ig); therefore, aflibercept only binds to VGEF-A / PIGF.

[0059] In certain specific (but non-limiting) embodiments, at least one small molecule anti-angiogenic agent specifically and physically interacts with one or more of VEGF-A, VEGF-B, VEGF-C and / or VEGF-D to inhibit VEGF / VEGFR interactions and VEGFR signaling.

[0060] In certain specific (but non-limiting) embodiments, at least one small molecule anti-angiogenic agent specifically and physically interacts with one or more of VEGFR-1, VEGFR-2 and / or VEGFR-3 to inhibit VEGF / VEGFR interactions and VEGF signaling. In certain specific (but non-limiting) embodiments, at least one small molecule anti-angiogenic agent is a type I, type II and / or type III VEGFR inhibitor. Type I VEGFR inhibitors, also known as ATP competitive inhibitors, produce hydrophobic interactions with the adenine region and form 1 to 3 hydrogen bonds with the surrounding residues of the receptor active site, thereby competing for binding to the active "DFG-in" conformation in the ATP binding pocket. Non-limiting examples of type I VEGFR inhibitors that can be used according to the present invention include pazopanib, axitinib, ponatinib, motesanib, etc. and any combination thereof. Type II VEGFR inhibitors are characterized by binding to the inactive "DFG-outward" conformation of the kinase and occupying a hydrophobic pocket adjacent to the ATP binding site. Non-limiting examples of type II VEGFR inhibitors that can be used according to the present invention include cabozantinib, lenvatinib, regorafenib, delitinib, etc., and any combination thereof. Type III inhibitors, also known as covalent inhibitors, can exert their pharmacological functions by irreversibly binding to cysteine ​​at a specific site of the kinase. Non-limiting examples of type III VEGFR inhibitors include vatalanib, etc.

[0061] Compositions including small molecule anti-angiogenic agents can be provided in any formulation known in the art or contemplated herein. In certain specific (but non-limiting) embodiments, the composition including at least one small molecule anti-angiogenic agent contains one or more pharmaceutically acceptable carriers (thus, the composition may also be referred to as a "pharmaceutical composition"). Non-limiting examples of suitable pharmaceutically acceptable carriers include water; saline; dextrose 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; liposomal carriers; buffer solutions such as sodium chloride, saline, phosphate-buffered saline and / or other substances that are physiologically acceptable and / or safe to use; diluents; excipients such as polyethylene glycol (PEG); or any combination thereof. Suitable pharmaceutically acceptable carriers for pharmaceutical preparations are described, for example, in Remington: The Science and Practice of Pharmacy, 23rd Edition (2020).

[0062] In certain specific (but non-limiting) embodiments, the composition comprising a small molecule anti-angiogenic agent may further include one or more additional active agents. Various active agents or small molecule anti-angiogenic agents used in combination with alternating electric fields are known in the art, and certain combination therapies are approved by the FDA or are currently being tested in clinical trials. Non-limiting examples of therapeutic agents that can be used in combination with small molecule anti-angiogenic agents according to the present invention include anti-PD-1 therapeutic agents, such as (but not limited to) Pembrolizumab, Tislelizumab, Nivolumab, and Cemiplimab; anti-PD-L1 therapeutic agents, such as atezolizumab, avelumab, and durvalumab; chemotherapeutic drugs, such as (but not limited to) paclitaxel, docetaxel, Ifosamide, etoposide, and sirolimus. Vepesid, gemcitabine, lomustine, albumin-paclitaxel, temozolomide and carboplatin; TKI inhibitors, such as (but not limited to) everolimus; mTOR inhibitors; Akt inhibitors; PI3K inhibitors; PARP inhibitors; FGF inhibitors; anti-LAB3 drugs; anti-CTLA-4 therapeutics; aromatase inhibitors, such as (but not limited to) letrozole; biologics, such as monoclonal antibodies (such as but not limited to denosumab and pembrolizumab); anti-VEGF antibodies or anti-VEGFR antibodies (such as but not limited to VEGFR-2 antibodies, such as (but not limited to) ramucirumab and DC101); etc. and any combination thereof.

[0063] In certain specific (but non-limiting) embodiments, the small molecule anti-angiogenic agent present in the composition is conjugated to another substance. For example, but not limited to, the small molecule anti-angiogenic agent can be conjugated to particles or other substances for targeted delivery of the drug to a specific location in the body. In another specific (but non-limiting) embodiment, the composition may include a small molecule anti-angiogenic agent encapsulated in nanoparticles.

[0064] In addition, any composition of the present invention may contain other agents that allow the composition to be administered via a specific route of administration. For example, but not limited to, the composition can be formulated for administration by oral, topical, transdermal, parenteral, subcutaneous, intranasal, mucosal, intramuscular, intraperitoneal, intravitreal and / or intravenous routes. Based on the route of administration, the composition may contain one or more additional components in addition to an active agent (e.g., a small molecule anti-angiogenic agent and / or an additional therapeutic agent). Examples of other second compounds that may be present include, but are not limited to, fillers, salts, buffers, preservatives, stabilizers, solubilizers, wetting agents, emulsifiers, dispersants, and other materials well known in the art.

[0065] In specific (but non-limiting) embodiments, at least one composition comprising a small molecule anti-angiogenic agent is orally administered to the cells / subject / tumor.

[0066] At least one composition including a small molecule anti-angiogenic agent can be administered before or after the application of the alternating electric field begins. In certain specific (but non-limiting) embodiments, at least one composition including a small molecule anti-angiogenic agent can be administered after the application of the alternating electric field begins. In particular (but not by way of limitation), at least one composition including a small molecule anti-angiogenic agent can be administered during the application of the alternating electric field (e.g., before the time period for applying the alternating electric field has passed) and / or after the application of the alternating electric field has passed.

[0067] For example, but not by way of limitation, at least one composition including a small molecule anti-angiogenic agent can be administered for a period of at least 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., and ranges formed by any of the above values ​​(e.g., a range of about 24 hours to about 96 hours, etc., and a range of about 14 hours to about 94 hours, etc.). In specific (but non-limiting) embodiments, at least one composition comprising a small molecule anti-angiogenic agent is administered at least about 24 hours after initiation of application of the alternating electric field.

[0068] In other non-limiting examples, at least one composition comprising a small molecule anti-angiogenic agent can be administered after the time period for which the alternating electric field is applied has elapsed, wherein the at least one composition comprising a small molecule anti-angiogenic agent is administered within 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. after the time period has elapsed.

[0069] In specific (but non-limiting) embodiments, at least one composition comprising a small molecule anti-angiogenic agent is administered within about 96 hours of the elapsed period of time.

[0070] Compositions including small molecule anti-angiogenic agents can be administered to cancer cells / subjects at any concentration that provides a therapeutically effective concentration of small molecule anti-angiogenic agents. In certain non-limiting embodiments, the application of an alternating electric field reduces the amount of small molecule anti-angiogenic agents required for therapeutic effectiveness compared to the normal therapeutically effective amount administered without an alternating electric field. For example, but not as a limitation, relative to the dose of a small molecule anti-angiogenic agent known to be therapeutically effective without the application of an alternating electric field, the therapeutically effective concentration of the small molecule anti-angiogenic agent can 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. In a specific (but non-limiting) embodiment, the therapeutically effective concentration of the small molecule anti-angiogenic agent is reduced by at least about 50% compared to the dose of a small molecule anti-angiogenic agent known to be therapeutically effective in the absence of an alternating electric field.

[0071] The therapeutically effective concentration of the small molecule anti-angiogenic agent used according to the present invention can be, for example, but not limited to, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 11 nM, about 12 nM, about 12.5 nM, about 13 nM, about 14 nM, about 15 nM, about 20 nM, about 25 nM, about 30 nM, about 35 nM, about 40 nM, about 45 nM, about 50 nM, about 55 nM. The present invention relates to a range of about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 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 1000 nM, about 1100 nM, about 1200 nM, about 1300 nM, about 1400 nM, about 1500 nM, etc., as well as ranges formed by any of the above values ​​(e.g., a range of about 12.5 nM to about 100 nM, etc.), and ranges combining two integers falling between two of the above values ​​(e.g., a range of about 170 nM to about 830 nM, etc.).

[0072] In specific (but non-limiting) embodiments, the therapeutically effective concentration of the small molecule anti-angiogenic agent is from about 12.5 nM to about 100 nM.

[0073] In certain specific (but non-limiting) embodiments, the method includes one or more additional steps. For example (but not by way of limitation), the method may further include the following steps: (3) stopping the application of the alternating electric field (for example but not limited to) to allow the cells / tissue to recover. In addition, any step (1) and / or (2) may be repeated one or more times.

[0074] In certain specific (but non-limiting) embodiments, the method includes concurrent therapy with two or more compositions. Thus, the method may include an additional step (4) of administering at least a second composition to the cancer cells / subject. In specific (but non-limiting) embodiments, at least the second composition may contain one or more of any active substances disclosed herein or otherwise contemplated for use with small molecule anti-angiogenic agents.

[0075] Various substances and therapies used in combination with small molecule anti-angiogenic agents are known in the art, and certain combination therapies are approved by the FDA or are currently being tested in clinical trials. Non-limiting examples of therapeutic agents that can be used in combination with small molecule anti-angiogenic agents according to the present invention include anti-PD-1 therapeutic agents, such as, but not limited to, Pembrolizumab, Tislelizumab, Nivolumab, and Cemiplimab; anti-PD-L1 therapeutic agents, such as atezolizumab, avelumab, and durvalumab; chemotherapeutic drugs, such as, but not limited to, paclitaxel, docetaxel, Ifosamide, etoposide, and sirolimus. Vepesid, gemcitabine, lomustine, albumin-paclitaxel, temozolomide and carboplatin; TKI inhibitors, such as (but not limited to) everolimus; mTOR inhibitors; Akt inhibitors; PI3K inhibitors; PARP inhibitors; FGF inhibitors; anti-LAB3 drugs; anti-CTLA-4 therapeutics; aromatase inhibitors, such as (but not limited to) letrozole; biologics, such as monoclonal antibodies (such as but not limited to denosumab and pembrolizumab); anti-VEGF or anti-VEGFR antibodies (such as but not limited to anti-VEGFR-2 antibodies, such as (but not limited to) ramucirumab and DC101); etc. and any combination thereof.

[0076] When present, step (4) can be performed substantially simultaneously or in whole or in part sequentially with the administration of the first composition in step (1), whereby the two separate compositions are administered simultaneously or in whole or in part sequentially. In addition, the two compositions administered in steps (1) and (4) can be administered by the same route (e.g., oral administration), or the two compositions can be administered by different routes (e.g., oral administration of one composition and intravenous administration of the other composition).

[0077] When present, the optional further application step (4) may be carried out before or after the start of application of the alternating electric field, during application of the alternating electric field, and / or after application of the alternating electric field has elapsed, in the same manner and within the same time frame as described above for the first composition.

[0078] That is, for example (but not by way of limitation), the second composition can be applied at least 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., and ranges formed by any of the above values ​​(e.g., a range of about 24 hours to about 96 hours, etc., and a combination of two integers falling between two of the above values, a range of about 14 hours to about 94 hours, etc.). In specific (but non-limiting) embodiments, the second composition is applied at least about 24 hours after initiation of application of the alternating electric field.

[0079] In other non-limiting examples, the second composition can be applied after the time period for applying the alternating electric field has passed, wherein the second composition is applied within 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. after the time period has passed. In a specific (but non-limiting) embodiment, the second composition is administered within about 96 hours after the elapsed time period.

[0080] In addition, for example (but not by way of limitation), the second composition can be applied at least about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 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 44 hours, or about 45 minutes after the first composition is applied. In some embodiments, the second composition is administered at least about 12 hours after the first composition is administered.

[0081] In certain specific (but non-limiting) embodiments, the method may further include the following steps: (5) administering at least one additional therapy to the cell / subject. The methods disclosed in the art are known to use any therapy known in the art or contemplated herein for use with alternating electric field (e.g., TTFields) and / or small molecule anti-angiogenic therapy. Non-limiting examples of other therapies that may be used include radiotherapy, photodynamic therapy, transarterial chemoembolization (TACE), or a combination thereof.

[0082] In certain specific (but non-limiting) embodiments, the method may further include the following steps: (6) applying a second alternating electric field to the target area of ​​the subject, wherein the alternating electric field is applied at a frequency different from the frequency of the alternating electric field of step (2). That is, the method of the present invention may include applying a first alternating electric field at a first frequency within a first time period, wherein applying a first alternating electric field at a first frequency within a first time period increases the permeability of the cell membrane of cancer cells; then at least one composition including at least one small molecule anti-angiogenic agent may be applied, and the increased cell membrane permeability enables the substance to pass through the cell membrane. Then a second alternating electric field may be applied at a second frequency for a second time period; the second frequency is different from the first frequency, and the second alternating electric field of the second frequency reduces the vitality of cancer cells. The use of alternating electric fields at two different frequencies (to affect cell membrane permeability and cell vitality) is discussed in detail in U.S. Patent Application Publication No. US2020 / 0009376, the entire contents of which are expressly incorporated herein by reference.

[0083] Any of steps (1) and (2) and optional steps (3), (4), (5) and (6) may be repeated one or more times. Each step may be repeated as many times as desired. When repeating step (2), the transducer array may be placed in a slightly different position on the subject than its original position; repositioning the array in this manner may further aid in treating the tumor / cancer. In addition, step (1) of administering the composition / additional therapy and optional steps (4) and (5) (when present) may be repeated different times and at different intervals to follow any known and / or generally accepted dosage / treatment regimen of the composition / therapy.

[0084] The use of sequential references to optional steps is for illustrative purposes only; the method of the present invention may include one or more of the optional steps (3), (4), (5), and (6), alone or in combination with one another. That is, the method of the present invention includes performing step (3) in the absence of step (4) or (5), performing step (4) in the absence of step (3) or (5), performing step (5) in the absence of steps (3) and (4), and performing (6) in the absence of (3), (4), and / or (5). In other words, the scope of the method disclosed herein includes performing steps (1)-(2) (and repeating each step a desired number of times), performing steps (1)-(3) (and repeating one or more of steps (1)-(3) a desired number of times), performing steps (1)-(2) and (4) (and repeating one or more of steps (1)-(2) and (4) a desired number of times), performing steps (1)-(2) and (5) (and repeating one or more of steps (1)-(2) and (5) a desired number of times); performing steps (1)-(4) (and repeating steps (1)-(4) (a) performing one or more of the steps (1)-(3) and (5) as many times as desired), performing steps (1)-(2) and (4)-(5) (and repeating one or more of the steps (1)-(2) and (4)-(5) as many times as desired); performing step (6) in combination with step (1)-(2) alone or in combination with one or more of steps (3)-(5), and performing all of steps (1)-(6) (and repeating one or more of steps (1)-(6) as many times as desired).

[0085] Although the above explicitly describes concurrent therapy using two substances, it should be understood that the scope of the present invention also includes concurrent therapy using three or more compositions. Therefore, the method may include one or more additional steps (similar to steps (1) and (4)) of administering additional compositions to the subject. Any additional substance administered in the method may be selected from any substance disclosed herein or otherwise expected to be used in combination with a small molecule anti-angiogenic agent (as disclosed above with respect to optional step (4)); in addition, the administration of any additional substance may be substantially simultaneous or in whole or in part sequential with the administration of the first and / or second composition / substance, and in the same manner and time range as the first and second compositions / substances described above.

[0086] Certain non-limiting embodiments of the present invention are directed to kits comprising any of the components of an alternating electric field (e.g., TTFields) generating system disclosed herein or otherwise contemplated (e.g., one or more transducer arrays and / or one or more hydrogel compositions, such as those described in U.S. Pat. Nos. 7,016,725; 7,089,054; 7,333,852; 7,565,205; 8,244,345; 8,715,203; 8,764,6 75; 10,188,851; 10,441,776 and 11,452,863; and in U.S. Patent Application Nos. US2018 / 0001078; US2018 / 0160933; US2019 / 0117956; US2019 / 0307781; and US2019 / 0308016) in combination with at least one of any compositions disclosed herein or otherwise considered including small molecule anti-angiogenic agents. The kit may optionally further include one or more of any optional compositions disclosed herein or otherwise considered (e.g., but not limited to, one or more optional compositions containing at least one additional active agent). The kit may optionally further include one or more devices (or one or more components of a device) used in one or more additional treatment steps.

[0087] In specific (but non-limiting) embodiments, the kit may further include instructions for performing any method disclosed herein or otherwise contemplated. For example (but not by way of limitation), the kit may include instructions for applying one or more components of an alternating electric field (e.g., TTFields) generating device to the patient's skin, instructions for applying the alternating electric field to the patient, instructions for when and how to apply a composition comprising a small molecule anti-angiogenic agent and optionally how to apply one or more optional additional compositions, and / or instructions for when to activate and deactivate the alternating electric field relative to the application of a composition comprising a small molecule anti-angiogenic agent and / or the application of one or more optional compositions and / or treatment steps.

[0088] In addition to the components described in detail above, the kit may also include other components / reagents for performing any specific method described herein or otherwise contemplated. For example (but not by way of limitation), the kit may further include: (i) components for preparing the skin prior to treatment with the hydrogel composition and / or the transducer array thereon (e.g., a razor, a cleaning composition or a 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) and / or (iv) other components for use with the system (i.e., conductive materials, non-conductive materials, emollient gels or creams, bandages, etc.). The properties of these additional components / reagents will depend on the specific treatment modality, and their identification is well within the skill of a person of ordinary skill in the art; therefore, no further description thereof is required. In addition, the components / reagents present in the kit may each be in a separate container / compartment, or the various components / reagents may be combined in one or more containers / compartments, depending on the sterility, cross-reactivity, and stability of the components / reagents.

[0089] The test kit can be arranged in any packaging that allows the components present therein to function according to the present invention. In certain non-limiting embodiments, the test kit further comprises a sealed package in which the components are placed. In certain specific (but non-limiting) embodiments, the sealed package is substantially airtight and / or substantially light-tight.

[0090] In addition, the kit may also include a set of written instructions for how to use one or more components of the kit. Kits of this nature can be used in any method described herein or otherwise contemplated.

[0091] In certain 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.

[0092] Certain non-limiting embodiments of the present invention are directed to systems that include any of the components of an alternating electric field generating system disclosed herein or otherwise contemplated (e.g., but not limited to, one or more transducer arrays and / or one or more hydrogel compositions, as described in U.S. Patents 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 11,452,863; and in U.S. Patent Applications Nos. US2018 / 0001078; US2018 / 0160933; US2019 / 0117956; US 2019 / 0307781; and US2019 / 0308016) in combination with at least one of any compositions disclosed herein or otherwise contemplated comprising a small molecule anti-angiogenic agent. The system may optionally further include one or more of any optional compositions disclosed herein or otherwise contemplated. The system may optionally further include one or more devices (or one or more components of a device) used in one or more additional treatment steps.

[0093] Examples

[0094] Examples are provided below. However, the present invention should be understood to be not limited in its application to the specific experiments, results and laboratory procedures disclosed below. On the contrary, the examples are provided as only one of the various embodiments and are exemplary, not exhaustive.

[0095] Example 1

[0096] In contrast to the prior art mentioned in the Background section above, which suggests that TTFields have anti-angiogenic activity, this example demonstrates that TTFields treatment increases the expression of VEGF and actually increases the formation of new vasculature.

[0097] An orthotopic ovarian cancer mouse model was generated by injecting mice with MOSE-L-FFL (day 0) and confirmed by IVIS on day 15. Starting from day 16, mice were treated with 200 kHz TTFields (or thermal control) for 10 days. Figure 1 As shown, increased vasculature was observed in tumor-filled ovaries of mice treated with TTFields when compared to untreated controls (heat-treated mice).

[0098] Mice from the in vivo model of ovarian cancer were sacrificed on day 26, and blood samples were drawn and collected in designated serum tubes, centrifuged at 1000 g for 15 min and kept at -20°C. The serum was incubated on the cytokine array membrane as Figure 2As shown, an increase in secreted VEGF was observed after TTFields treatment compared to heat-treated mice.

[0099] Next, mice with orthotopic lung cancer were generated by injecting them with LLC-2 (day 0) and confirmed on day 7. Starting from day 7, mice were treated with 150 kHz TTFields for 14 days. On day 21, mice were sacrificed, blood samples were drawn and collected in designated serum tubes, centrifuged at 1000 g for 15 minutes and kept at -20°C. Serum was incubated on cytokine array membranes as Figure 3 As shown, an increase in VEGF secretion was observed after TTFields compared to heat-treated mice.

[0100] Next, on the 26th day, the Figure 1-2 The same in vivo model of ovarian cancer was used in mice, and tumor samples were taken for histological analysis of tumor tissue. Tumor tissue was stained for CD31 (a marker of endothelial cells and vascular system) by immunohistochemistry, and the images were quantified. CD31 staining is represented by brown (DAB staining). Figure 4 As shown, the microvessel density of TTFields-treated tumors was significantly higher than that of heat-treated control tumors.

[0101] Thus, contrary to the current literature indicating that TTFields therapy has an anti-angiogenic effect, this example demonstrates that administration of TTFields actually has a pro-angiogenic effect, which results in increased VEGF expression and formation of new vasculature to feed the tumor. Thus, administration of at least one small molecule anti-angiogenic agent counteracts the pro-angiogenic effect of TTFields administration in the combination therapy.

[0102] Example 2

[0103] Lenvatinib (prescription name: Eisai Inc. Nutley, NJ) is a multikinase inhibitor of VEGFR1-3. It is clinically used to treat differentiated thyroid cancer (DTC), advanced renal cell carcinoma (RCC) and hepatocellular carcinoma.

[0104] This example examines the combined effects of TTFields and lenvatinib in an attempt to understand the impact of the tumor microenvironment and the changes that the extracellular matrix (ECM) undergoes after application of TTFields.

[0105] The tumor microenvironment, particularly the ECM components, has been shown to be a critical part of tumor progression and clinical outcome (Henke et al. (2020) Front Mol Biosci. 6:160; Baghban et al. (2020) Cell Commun Signal, 18:59; Popova et al. (2022) Cancers, 14(1):238). Lenvatinib is a multikinase inhibitor that inhibits many of the pathways we have observed to be upregulated after TTFields application: the three major vascular endothelial growth factor receptors 1, 2, and 3 (VEGFR-1, -2, and -3); fibroblast growth factor receptors 1, 2, 3, and 4 (FGFR-1, -2, -3, and -4); platelet-derived growth factor receptor alpha (PDGFRα); c-Kit; and the RET proto-oncogene. Furthermore, these pathways are known to regulate ECM formation (Nelson et al. (1997) J Vasc Surg, 26(1): 104-12; Xu et al. (1996) J Cell Biol, 132(1-2): 239-49; Vlodavsky et al. (1990) Cancer Metastasis Rev, 9(3): 203-2).

[0106] This case study studies TTFields and lenvatinib in INOVITRO TM Combination therapy with collagen and cancer cells grown as 3D structures in an INOVIVO 3D model (Novocure GmbH, Root, Switzerland). TM Combination therapy in the system (Novocure GmbH, Root, Switzerland; see, e.g., Davidi (2020) Neuro-Oncology, 22(Suppl 2):ii104).

[0107] Perform experiments to assess cell and tumor growth (cell counting or 3D image capture INOVITRO TM , tumor growth INOVIVO TM Monitoring), apoptosis (INOVITRO TM -Annexin V / PI staining and INOVIVO TM - Ki67 tissue staining), secretion of cytokines and chemokines (in tumor-derived lysates as well as in conditioned media and serum examined by ELISA, cytokine arrays, multiplex assays or luminex assays), collagen expression (by collagen staining and secretion) and vascularization of tissues - angiogenesis (by CD31 staining).

[0108] The cell lines used are for cancers typically treated with lenvatinib (such as, but not limited to, differentiated thyroid carcinoma (DTC), advanced renal cell carcinoma (RCC) and / or hepatocellular carcinoma) and / or cancer cell lines typically treated with TTFields (such as, but not limited to, glioblastoma or pleural mesothelioma).

[0109] Specifically, the combined effect of TTFields and lenvatinib was examined in an ovarian cancer cell line (A2780). The effect of combined treatment on cell counts and apoptosis was Figure 5-6 As can be seen, the combination of TTFields and lenvatinib provides an enhanced effect over either therapy alone.

[0110] Thus, there is a synergistic effect using a combination therapy including TTFields and lenvatinib. The combined TTFields / lenvatinib therapy is more effective in reducing the viability of cancer cells and reducing tumor volume and / or preventing tumor volume from increasing when compared to either treatment alone. In addition, the use of TTFields in combination with lenvatinib reduces the amount of lenvatinib required to produce a therapeutic result; when lenvatinib is combined with TTFields, the therapeutically effective concentration of lenvatinib is reduced by at least about 50% compared to a dose of lenvatinib known to be therapeutically effective in the absence of an alternating electric field.

[0111] Non-limiting illustrative embodiments of the present inventive concept

[0112] Illustrative Example 1. A method for reducing the viability of cancer cells, the method comprising the steps of: (1) administering to the cancer cells at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, the anti-angiogenic agent having an IC of less than about 10 nmol / L. 50 specifically [physically] interact with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibit the interaction between VEGF and VEGFR and / or inhibit VEGFR signal transduction; and (2) applying an alternating electric field to the cancer cells for a period of time.

[0113] Illustrative Example 2. The method of Illustrative Example 1, wherein the method is performed in vitro or ex vivo.

[0114] Illustrative Example 3. The method of Illustrative Example 1, wherein the method is performed in vivo.

[0115] Illustrative embodiment 4. The method of any one of illustrative embodiments 1-3, 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, pancreatic cancer, lung cancer cells, breast cancer cells, and combinations thereof.

[0116] Illustrative Example 5. A method for treating cancer in a subject, the method comprising the steps of: (1) administering to the subject at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, the anti-angiogenic agent having an IC of less than about 10 nmol / L. 50 Specifically interacting with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibiting the interaction between VEGF and VEGFR and / or inhibiting VEGFR signal transduction; and (2) applying an alternating electric field to a target area of ​​a subject.

[0117] Illustrative Example 6. A method of reducing tumor volume and / or preventing tumor volume increase, wherein the tumor is present in a living subject and comprises a plurality of cancer cells, the method comprising the following steps: (1) administering to the subject at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, the anti-angiogenic agent having an IC of less than about 10 nmol / L; 50 Specifically interacting with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibiting the interaction between VEGF and VEGFR and / or inhibiting VEGFR signal transduction; and (2) applying an alternating electric field to a target area of ​​a subject.

[0118] Illustrative Example 7. The method of Illustrative Example 5 or 6, wherein at least one composition is orally administered to the subject.

[0119] Illustrative embodiment 8. The method of any of illustrative embodiments 5-7, wherein the cancer is selected from the group consisting of hepatocellular carcinoma, glioblastoma, pleural mesothelioma, differentiated thyroid cancer, advanced renal cell carcinoma, ovarian cancer, pancreatic cancer, lung cancer, breast cancer, and combinations thereof.

[0120] Illustrative Example 9. The method of any one of Illustrative Examples 1-8, wherein at least one small molecule anti-angiogenic agent specifically interacts with at least one of VEGF-A, VEGF-B, VEGF-C, or VEGF-D.

[0121] Illustrative Example 10. The method of any one of Illustrative Examples 1-9, wherein at least one small molecule anti-angiogenic agent specifically interacts with at least one of VEGFR-1, VEGFR-2, or VEGFR-3.

[0122] Illustrative embodiment 11. The method of any one of illustrative embodiments 1-10, wherein at least one small molecule anti-angiogenic agent is selected from the group consisting of lenvatinib, axitinib, regorafenib, cabozantinib, anlotinib, pazopanib, albendazole, delitinib, motesanib, aflibercept, ponatinib, cediranib, tivozanib, telatinib, apatinib, sematinib, fruquintinib, delitinib, anlotinib, taxifolin, surufatinib, dovitinib, ningetinib, AZD2932, LY2874455, MGCD-265 analogs, ZM 306416, ZM 323881, KRN 633, YF-452, ODM-203, AEE 788, BMS 605541, MAZ51, Ki 8751, SU 5402, SU 5408, SU5205, SU5214, SU6668, SU 14813, XL 092, XL 184, BAW2881, BFH772, A-13958, SKLB1002, WAY-340935, ZD-4190, hVEGF-IN-1, R1530, VEGF-Grab, soluble vascular endothelial growth factor decoy receptor FP3, VEGF decoy receptor fusion protein, VEGF decoy receptor and combinations thereof.

[0123] Illustrative Example 12. The method of any one of Illustrative Examples 1-11, wherein at least one small molecule anti-angiogenic agent is a type I VEGFR inhibitor.

[0124] Illustrative Example 13. The method of Illustrative Example 12, wherein the type I VEGFR inhibitor is selected from the group consisting of pazopanib, axitinib, ponatinib, motesanib, and combinations thereof.

[0125] Illustrative Example 14. The method of any one of Illustrative Examples 1-13, wherein at least one small molecule anti-angiogenic agent is a type II VEGFR inhibitor.

[0126] Illustrative Example 15. The method of Illustrative Example 14, wherein the type II VEGFR inhibitor is selected from the group consisting of cabozantinib, lenvatinib, regorafenib, delimtinib, and combinations thereof.

[0127] Illustrative Example 16. The method of any one of Illustrative Examples 1-15, wherein at least one small molecule anti-angiogenic agent is a type III VEGFR inhibitor.

[0128] Illustrative Example 17. A method for reducing cancer cell viability, and / or treating cancer in a subject, and / or reducing tumor volume and / or preventing tumor volume increase, the method comprising the following steps: (1) administering at least one composition to the cancer cells / subject, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, which is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR), which specifically and exclusively binds to VEGF or VEGFR and does not bind to other targets; and (2) applying an alternating electric field to the cancer cell / target area of ​​the subject for a period of time.

[0129] Illustrative embodiment 18. The method of illustrative embodiment 17, wherein the method is performed in vitro or ex vivo.

[0130] Illustrative Example 19. The method of Illustrative Example 17, wherein the method is performed in vivo.

[0131] Illustrative embodiment 20. The method of any one of illustrative embodiments 17-19, wherein the cancer / cancer cell is selected from the group consisting of hepatocellular carcinoma cells, glioblastoma cells, pleural mesothelioma cells, differentiated thyroid carcinoma cells, advanced renal cell carcinoma cells, ovarian cancer, pancreatic cancer, lung cancer cells, breast cancer cells, and combinations thereof.

[0132] Illustrative embodiment 21. The method of any one of illustrative embodiments 17-20, wherein at least one small molecule anti-angiogenic agent is aflibercept.

[0133] Illustrative Example 22. A method of reducing the viability of cancer cells, the method comprising the steps of: (1) administering at least one composition to the cancer cells, wherein the at least one composition comprises lenvatinib; and (2) applying an alternating electric field to the cancer cells for a period of time.

[0134] Illustrative embodiment 23. The method of illustrative embodiment 22, wherein the method is performed in vitro or ex vivo.

[0135] Illustrative embodiment 24. The method of illustrative embodiment 22, wherein the method is performed in vivo.

[0136] Illustrative embodiment 25. The method of any one of illustrative embodiments 22-24, wherein the cancer / cancer cell is selected from the group consisting of hepatocellular carcinoma cells, glioblastoma cells, pleural mesothelioma cells, differentiated thyroid carcinoma cells, advanced renal cell carcinoma cells, ovarian cancer, pancreatic cancer, lung cancer cells, breast cancer cells, and combinations thereof.

[0137] Illustrative Example 26. A method of treating cancer in a subject, the method comprising the steps of: (1) administering to the subject at least one composition, wherein the at least one composition comprises lenvatinib; and (2) applying an alternating electric field to a target area of ​​the subject.

[0138] Illustrative Example 27. A method for reducing tumor volume and / or a method for preventing an increase in tumor volume, wherein the tumor is present in the body of a living subject and comprises a plurality of cancer cells, the method comprising the steps of: (1) administering to the subject at least one composition, wherein the at least one composition comprises lenvatinib; and (2) applying an alternating electric field to a target area of ​​the subject.

[0139] Illustrative Embodiment 28. The method of Illustrative Embodiment 26 or 27, wherein at least one composition is orally administered to the subject.

[0140] Illustrative Example 29. The method of any one of Illustrative Examples 26-28, wherein the composition comprising lenvatinib is administered to the cancer cells at a therapeutically effective concentration of lenvatinib.

[0141] Illustrative embodiment 30. The method of illustrative embodiment 29, wherein the therapeutically effective concentration of lenvatinib is reduced by at least about 50% relative to a dose of lenvatinib known to be therapeutically effective in the absence of the alternating electric field.

[0142] Illustrative embodiment 31. The method of illustrative embodiment 29 or 30, wherein the therapeutically effective concentration of lenvatinib is about 12.5 nM to about 100 nM.

[0143] Illustrative embodiment 32. A method as in any of illustrative embodiments 1-31, wherein at least one of the following: 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 a field strength of at least about 1 V / cm in at least a portion of the cancer cells; and the alternating electric field is applied for a period of at least about 24 hours.

[0144] Illustrative Embodiment 33. The method of any of Illustrative Embodiments 1-32, wherein steps (1) and (2) are performed substantially simultaneously.

[0145] Illustrative embodiment 34. The method of any of illustrative embodiments 1-32, wherein steps (1) and (2) are performed completely or partially sequentially, and wherein at least one composition is applied before initiating application of the alternating electric field.

[0146] Illustrative Embodiment 35. The method of any of Illustrative Embodiments 1-33, wherein steps (1) and (2) are performed completely or partially sequentially, and wherein at least one composition is applied after application of the alternating electric field has begun.

[0147] Illustrative Embodiment 36. The method of Illustrative Embodiment 35, wherein the at least one composition is applied at least 24 hours after application of the alternating electric field has begun.

[0148] Illustrative Embodiment 37. The method of Illustrative Embodiment 36, wherein the at least one composition is applied before the period of time during which the alternating electric field is applied elapses.

[0149] Illustrative Embodiment 38. The method of Illustrative Embodiment 36, wherein the at least one composition is administered after the time period has elapsed.

[0150] Illustrative Embodiment 39. The method of Illustrative Embodiment 38, wherein the at least one composition is administered within about 96 hours after the time period has elapsed.

[0151] Illustrative Embodiment 40. The method of any of Illustrative Embodiments 1-39, wherein the time period for which the alternating electric field is applied is in the range of from about 24 hours to about 72 hours.

[0152] Illustrative Embodiment 41. The method of any of Illustrative Embodiments 1-40, wherein the method further comprises the step of stopping the application of the alternating electric field.

[0153] Illustrative Embodiment 42. The method of any one of Illustrative Embodiments 1-41, wherein at least one composition further comprises a pharmaceutically acceptable carrier.

[0154] Illustrative embodiment 43. The method of any of illustrative embodiments 1-42, wherein steps (1) and (2) are repeated one or more times.

[0155] Illustrative Example 44. A method as described in any of Illustrative Examples 1-43, wherein the method includes the steps of applying a first alternating electric field to the cancer cells / target area of ​​the subject for a period of time and applying a second alternating electric field to the cancer cells / target area of ​​the subject for a period of time, and wherein the first and second alternating electric fields have different frequencies.

[0156] Illustrative embodiment 45. The method of illustrative embodiment 44, wherein the first frequency affects cell permeability, and wherein the second frequency affects cell viability.

[0157] Illustrative Embodiment 46. The method of any one of Illustrative Embodiments 1-45, wherein at least one composition further comprises at least one additional therapeutic agent.

[0158] Illustrative embodiment 47. The method of any one of illustrative embodiments 1-46, wherein the method further comprises the step of administering a second composition to the cancer cells, wherein the second composition comprises at least one additional therapeutic agent, and wherein the first and second compositions are administered substantially simultaneously or completely or partially sequentially.

[0159] Illustrative embodiment 48. The method of illustrative embodiment 46 or 47, wherein at least one additional therapeutic agent is selected from the group consisting of: a checkpoint immunosuppressant, an anti-PD-1 therapeutic agent, a chemotherapeutic agent, a TKI inhibitor, an mTOR inhibitor, an Akt inhibitor, an aromatase inhibitor, a radiotherapeutic agent, a biologic agent, and a combination thereof.

[0160] Illustrative embodiment 49. The method of illustrative embodiment 48, wherein the at least one additional therapeutic agent is selected from the group consisting of: pembrolizumab, tislelizumab, paclitaxel, docetaxel, Ifosamide, etoposide (Vepesid), gemcitabine, carboplatin, everolimus, letrozole, denosumab, and combinations thereof.

[0161] Illustrative embodiment 50. The method of any of illustrative embodiments 1-49, further comprising the step of administering to the cells / subject at least one additional therapy.

[0162] Illustrative Embodiment 51. The method of Illustrative Embodiment 50, wherein the at least one additional therapy is selected from the group consisting of radiation therapy, photodynamic therapy, transarterial chemoembolization (TACE), and combinations thereof.

[0163] Illustrative Example 52. A composition for use in the method of any of Illustrative Examples 1-16 and 32-51 (when relying on Illustrative Examples 1-16), wherein the composition comprises at least one small molecule anti-angiogenic agent having an IC of less than about 10 nmol / L. 50 It specifically interacts with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits VEGFR signal transduction.

[0164] Illustrative Example 53. Use of a composition in the method of any of Illustrative Examples 1-16 and 32-51 (when relying on Illustrative Examples 1-16), wherein the composition comprises at least one small molecule anti-angiogenic agent having an IC of less than about 10 nmol / L. 50It specifically interacts with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits VEGFR signal transduction.

[0165] Illustrative embodiment 54. A composition for use in the method of any of illustrative embodiments 17-21 and 32-51 (when dependent upon illustrative embodiments 17-21), wherein the composition comprises at least one small molecule anti-angiogenic agent that is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) that specifically and exclusively binds to VEGF or VEGFR and does not bind to other targets.

[0166] Illustrative Example 55. Use of a composition in the method of any of Illustrative Examples 17-21 and 32-51 (when dependent upon Illustrative Examples 17-21), wherein the composition comprises at least one small molecule anti-angiogenic agent that is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) that specifically and exclusively binds to VEGF or VEGFR and does not bind to other targets.

[0167] Illustrative Embodiment 56. A composition for use in the method of any of Illustrative Embodiments 22-31 and 32-51 (when relying upon Illustrative Embodiments 22-31), wherein the composition comprises lenvatinib.

[0168] Illustrative Embodiment 57. Use of a composition in the method of any of Illustrative Embodiments 22-31 and 32-51 (when relying upon Illustrative Embodiments 22-31), wherein the composition comprises lenvatinib.

[0169] Illustrative embodiment 58. A kit comprising: at least one pair of transducer arrays for generating an alternating electric field therebetween when the transducer arrays are applied to at least one cell and / or when the transducer arrays are placed on a subject; and at least one composition of any one of illustrative embodiments 52, 54 or 56.

[0170] Although the attached disclosure describes the concept of the present invention in conjunction with the specific experiments, results and language set forth below, it is apparent that many substitutions, modifications and variations are apparent to those skilled in the art. Therefore, it is intended to encompass all these substitutions, modifications and variations that fall within the spirit and broad scope of the present invention.

Claims

1. A kit comprising: at least one pair of transducer arrays for generating an alternating electric field therebetween when the transducer arrays are applied to at least one cell and / or when the transducer arrays are placed on a subject; as well as At least one composition comprising at least one small molecule anti-angiogenic agent having an IC of less than about 10 nmol / L 50 It specifically interacts with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits VEGFR signal transduction.

2. A kit comprising: at least one pair of transducer arrays for generating an alternating electric field therebetween when the transducer arrays are applied to at least one cell and / or when the transducer arrays are placed on a subject; as well as At least one composition comprising at least one small molecule anti-angiogenic agent that is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) that specifically and exclusively binds to VEGF or VEGFR and does not bind to other targets.

3. A kit comprising: at least one pair of transducer arrays for generating an alternating electric field therebetween when the transducer arrays are applied to at least one cell and / or when the transducer arrays are placed on a subject; as well as At least one composition comprising lenvatinib.

4. A composition for use in the method according to any one of claims 10-25 and 41-60, wherein The composition includes at least one small molecule anti-angiogenic agent, wherein the small molecule anti-angiogenic agent has an IC value of less than about 10 nmol / L. 50 It specifically interacts with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits VEGFR signal transduction.

5. Use of a composition in a method according to any one of claims 10-25 and 41-60, wherein The composition includes at least one small molecule anti-angiogenic agent, which specifically interacts with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) with an IC50 of less than about 10 nmol / L and selectively inhibits the interaction between VEGF and VEGFR and / or inhibits VEGFR signal transduction.

6. A composition for use in a method according to any one of claims 26-30 and 41-60, wherein The composition includes at least one small molecule anti-angiogenic agent that is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR), which specifically and exclusively binds to VEGF or VEGFR and does not bind to other targets.

7. Use of a composition in a method according to any one of claims 26-30 and 41-60, wherein The composition includes at least one small molecule anti-angiogenic agent that is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR), which specifically and exclusively binds to VEGF or VEGFR and does not bind to other targets.

8. A composition for use in a method according to any one of claims 31-60, wherein The composition includes lenvatinib.

9. Use of a composition in a method according to any one of claims 31 to 60, wherein The composition includes lenvatinib.

10. A method for reducing the viability of cancer cells, the method comprising the following steps: (1) administering at least one composition to the cancer cells, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, the anti-angiogenic agent having an IC of less than about 10 nmol / L 50 Specifically interacting with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibiting the interaction between VEGF and VEGFR and / or inhibiting VEGFR signal transduction; and (2) applying an alternating electric field to the cancer cells for a period of time.

11. The method according to claim 10, wherein: The methods are performed in vitro or ex vivo.

12. The method according to claim 10, wherein: The method is performed in vivo.

13. The method according to any one of claims 10 to 12, 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, pancreatic cancer, lung cancer cells, breast cancer cells, and combinations thereof.

14. A method for treating cancer in a subject, the method comprising the steps of: (1) administering to the subject at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, wherein the anti-angiogenic agent has an IC of less than about 10 nmol / L 50 Specifically interacting with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibiting the interaction between VEGF and VEGFR and / or inhibiting VEGFR signal transduction; and (2) Applying an alternating electric field to a target area of ​​the subject.

15. A method for reducing tumor volume and / or preventing said tumor volume from increasing, wherein: The tumor is present in the body of a living subject and includes a plurality of cancer cells, and the method comprises the following steps: (1) administering to the subject at least one composition, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, wherein the anti-angiogenic agent has an IC of less than about 10 nmol / L 50 Specifically interacting with vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR) and selectively inhibiting the interaction between VEGF and VEGFR and / or inhibiting VEGFR signal transduction; and (2) Applying an alternating electric field to a target area of ​​the subject.

16. The method according to claim 14 or 15, wherein: The at least one composition is orally administered to the subject.

17. The method according to any one of claims 14 to 16, wherein: The cancer cell is selected from the group consisting of hepatocellular carcinoma, glioblastoma, pleural mesothelioma, differentiated thyroid cancer, advanced renal cell carcinoma, ovarian cancer, pancreatic cancer, lung cancer, breast cancer, and combinations thereof.

18. The method according to any one of claims 10 to 17, wherein: The at least one small molecule anti-angiogenic agent specifically interacts with at least one of VEGF-A, VEGF-B, VEGF-C, or VEGF-D.

19. The method according to any one of claims 10 to 18, wherein: The at least one small molecule anti-angiogenic agent specifically interacts with at least one of VEGFR-1, VEGFR-2 or VEGFR-3.

20. The method according to any one of claims 10 to 19, wherein: The at least one small molecule anti-angiogenic agent is selected from the group consisting of lenvatinib, axitinib, regorafenib, cabozantinib, anlotinib, pazopanib, albendazole, delitinib, motesanib, aflibercept, ponatinib, cediranib, tivozanib, telatinib, apatinib, sematinib, fruquintinib, delitinib, anlotinib, tadalafil, surufatinib, dovitinib, ningetinib, AZD2932, LY2874455, MGCD-265 analogs, ZM 306416, ZM 323881, KRN 633, YF-452, ODM-203, AEE 788, BMS 605541, MAZ51, Ki 8751, SU 5402, SU 5408, SU5205, SU5214, SU 6668, SU14813, XL 092, XL 184, BAW2881, BFH772, A-13958, SKLB1002, WAY-340935, ZD-4190, hVEGF-IN-1, R1530, VEGF-Grab, soluble vascular endothelial growth factor decoy receptor FP3, VEGF decoy receptor fusion protein, VEGF decoy receptor and combinations thereof.

21. The method according to any one of claims 10 to 20, wherein: The at least one small molecule anti-angiogenic agent is a type I VEGFR inhibitor.

22. The method according to claim 21, wherein: The type I VEGFR inhibitor is selected from the group consisting of: pazopanib, axitinib, ponatinib, motesanib.

23. The method according to any one of claims 10 to 22, wherein: The at least one small molecule anti-angiogenic agent is a type II VEGFR inhibitor.

24. The method according to claim 23, wherein: The type II VEGFR inhibitor is selected from the group consisting of cabozantinib, lenvatinib, regorafenib, derafenib, and combinations thereof.

25. The method according to any one of claims 10 to 24, wherein: The at least one small molecule anti-angiogenic agent is a type III VEGFR inhibitor.

26. A method for reducing cancer cell viability, and / or treating cancer in a subject, and / or reducing tumor volume and / or preventing tumor volume increase, the method comprising the following steps: (1) administering at least one composition to the cancer cell / subject, wherein the at least one composition comprises at least one small molecule anti-angiogenic agent, which is a selective inhibitor of vascular endothelial growth factor (VEGF) or VEGF receptor (VEGFR), which specifically and solely binds to VEGF or VEGFR and does not bind to other targets; and (2) applying an alternating electric field to the cancer cell / target area of ​​the subject for a period of time.

27. The method according to claim 26, wherein: The methods are performed in vitro or ex vivo.

28. The method according to claim 26, wherein: The method is performed in vivo.

29. The method according to any one of claims 26 to 28, wherein: The cancer / cancer cell is selected from the group consisting of hepatocellular carcinoma cells, glioblastoma cells, pleural mesothelioma cells, differentiated thyroid carcinoma cells, advanced renal cell carcinoma cells, ovarian cancer, pancreatic cancer, lung cancer cells, breast cancer cells, and combinations thereof.

30. The method of any one of illustrative embodiments 26-29, wherein: The at least one small molecule anti-angiogenic agent is aflibercept.

31. A method for reducing the viability of cancer cells, the method comprising the following steps: (1) administering at least one composition to the cancer cells, wherein the at least one composition comprises lenvatinib; as well as (2) applying an alternating electric field to the cancer cells for a period of time.

32. The method according to claim 31, wherein: The methods are performed in vitro or ex vivo.

33. The method according to claim 31, wherein: The method is performed in vivo.

34. The method according to any one of claims 31 to 33, wherein: The cancer / cancer cell is selected from the group consisting of hepatocellular carcinoma cells, glioblastoma cells, pleural mesothelioma cells, differentiated thyroid carcinoma cells, advanced renal cell carcinoma cells, ovarian cancer, pancreatic cancer, lung cancer cells, breast cancer cells, and combinations thereof.

35. A method for treating cancer in a subject, the method comprising the steps of: (1) administering at least one composition to the subject, wherein the at least one composition comprises lenvatinib; as well as (2) Applying an alternating electric field to a target area of ​​the subject.

36. A method for reducing tumor volume and / or a method for preventing tumor volume increase, wherein: The tumor is present in the body of a living subject and includes a plurality of cancer cells, and the method comprises the following steps: (1) administering at least one composition to the subject, wherein the at least one composition comprises lenvatinib; and (2) Applying an alternating electric field to a target area of ​​the subject.

37. The method according to claim 35 or 36, wherein: The at least one composition is orally administered to the subject.

38. The method according to any one of claims 35 to 37, wherein: The composition comprising lenvatinib is administered to the cancer cells at a therapeutically effective concentration of lenvatinib.

39. The method of claim 38, wherein: The therapeutically effective concentration of lenvatinib is reduced by at least about 50% relative to a dose of lenvatinib known to be therapeutically effective in the absence of the alternating electric field.

40. The method according to claim 38 or 39, wherein: The therapeutically effective concentration of lenvatinib is about 12.5 nM to about 100 nM.

41. The method according to any one of claims 10 to 40, wherein: At least one of the following: applying an alternating electric field at a frequency in the range of about 50 kHz to about 1 MHz; The alternating electric field has a field strength of at least about 1 V / cm in at least a portion of the cancer cells; and The period of time during which the alternating electric field is applied is at least about 24 hours.

42. The method according to any one of claims 10 to 41, wherein: Steps (1) and (2) are performed substantially simultaneously.

43. The method according to any one of claims 10 to 41, wherein: Steps (1) and (2) are carried out in whole or in part sequentially, and wherein at least one composition is applied before starting to apply the alternating electric field.

44. The method according to any one of claims 10 to 41, wherein: Steps (1) and (2) are performed in whole or in part sequentially, and wherein at least one composition is applied after the start of application of the alternating electric field.

45. The method of claim 44, wherein: At least one composition is applied at least 24 hours after the start of application of said alternating electric field.

46. ​​The method of claim 45, wherein: The at least one composition is applied before the time for applying the alternating electric field has elapsed.

47. The method of claim 45, wherein: The at least one composition is administered after a period of time has passed.

48. The method of claim 47, wherein: The at least one composition is administered within about 96 hours after the period of time has elapsed.

49. The method according to any one of claims 10 to 48, wherein: The time period for applying the alternating electric field is from about 24 hours to about 72 hours.

50. The method according to any one of claims 10 to 49, wherein: The method further comprises the step of ceasing said applying of said alternating electric field.

51. The method according to any one of claims 10 to 50, wherein: The at least one composition further comprises a pharmaceutically acceptable carrier.

52. The method according to any one of claims 10 to 51, wherein: Steps (1) and (2) are repeated one or more times.

53. The method according to any one of claims 10 to 52, wherein: The method includes the steps of applying a first alternating electric field to the cancer cell / target area of ​​the subject for a period of time and applying a second alternating electric field to the cancer cell / target area of ​​the subject for a period of time, and wherein the first and second alternating electric fields have different frequencies.

54. The method of claim 53, wherein: The first frequency affects cell permeability, and wherein the second frequency affects cell viability.

55. The method according to any one of claims 10 to 54, wherein: The at least one composition further comprises at least one additional therapeutic agent.

56. The method according to any one of claims 10 to 55, wherein: The method further comprises the step of administering to the cancer cells a second composition, wherein the second composition comprises at least one additional therapeutic agent, and wherein the first and second compositions are administered substantially simultaneously or in whole or in part sequentially.

57. The method of claim 55 or 56, wherein: The at least one additional therapeutic agent is selected from the group consisting of a checkpoint immunosuppressant, an anti-PD-1 therapeutic agent, a chemotherapeutic agent, a TKI inhibitor, an mTOR inhibitor, an Akt inhibitor, an aromatase inhibitor, a radiotherapeutic agent, a biologic agent, and combinations thereof.

58. The method of claim 57, wherein: The at least one additional therapeutic agent is selected from the group consisting of pembrolizumab, tislelizumab, paclitaxel, docetaxel, Ifosamide, etoposide (Vepesid), gemcitabine, carboplatin, everolimus, letrozole, denosumab, and combinations thereof.

59. The method of any one of claims 10-58, further comprising the step of administering to the cell / subject at least one additional therapy.

60. The method of claim 59, wherein: The at least one additional therapy is selected from the group consisting of radiation therapy, photodynamic therapy, transarterial chemoembolization (TACE), and combinations thereof.

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