Compositions, systems, and methods for treating cancer, tumor treatment fields, and anti-VEGFR-2 antibodies

Abstract

Disclosed are compositions, systems, and methods for reducing cancer cell viability, treating cancer, and preventing tumor growth within a living body. The systems and methods involve the application of an alternating electric field in combination with the administration of at least one composition comprising at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (anti-VEGFR-2 antibody).

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 477,396, filed December 28, 2022. The entire contents of the above-referenced patent application(s) are expressly incorporated herein by reference.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT Not applicable [Background technology]

[0003] Tumor treating fields (TT fields) are low-intensity (e.g., 1-3 V / cm) alternating electric fields in the mid-frequency range (e.g., 100-500 kHz) that target solid tumors by inhibiting mitosis. This non-invasive treatment is targeted to solid tumors and is described, for example, in U.S. Patent Nos. 7,016,725, 7,089,054, 7,333,852, 7,565,205, 8,244,345, 8,715,203, 8,764,675, 10,188,851, and 10,441,776. TT fields are typically delivered through two pairs of transducer arrays, each consisting of an electrode array positioned on opposite sides of the body part being treated, that generate perpendicular fields within the tumor being treated. Specifically, in the OPTUNE® system, one pair of electrodes is placed on the left and right (LR) sides of the tumor, and another pair is placed on the anterior-posterior (AP) sides of the tumor. TT fields are approved for the treatment of glioblastoma multiforme (GBM) and can be delivered, for example, via the OPTUNE® system (Novocure Limited, St. Helier, Jersey), which includes a transducer array placed on the patient's shaved head.

[0004] In the OPTUNE® device, each transducer array used to deliver the TT field consists of a series of ceramic disc electrodes coupled to the patient's skin (e.g., a patient's shaved head for the treatment of GBM) via a layer of conductive medical gel. The purpose of the medical gel is to conform to the contours of the body and provide good electrical contact between the array and the skin, allowing the gel interface to bridge the skin and reduce interference. The device is intended for continuous wear by the patient for 2–4 days, after which it is removed for hygiene and re-shaving (if necessary), and a new set of arrays is reapplied. Thus, the medical gel remains in substantially continuous contact with an area of ​​the patient's skin for 2–4 days at a time. Additionally, the array can be shifted by a few centimeters in either direction, allowing the skin to heal between treatment periods. Thus, an area of ​​skin covered by electrodes / gel for 2–4 days is exposed for 2–4 days by slightly shifting the replacement electrodes, after which the device can be reapplied to the original area of ​​skin for the next 2–4 days.

[0005] Angiogenesis is the process of forming new blood vessels to increase the supply of nutrients and oxygen to areas of the body that lack blood supply. Angiogenesis has been shown to be one of the hallmark processes of cancer. In addition, vascular endothelial growth factor (VEGF) has been shown to play an important role in promoting angiogenesis.

[0006] Investigating TT fields as a potential therapeutic improvement for osteosarcoma, Oh et al. (Technology in Cancer Research & Treatment (2020) doi:10.1177 / 1533033820947481) demonstrated that TT fields prevented angiogenesis in human tumor endothelial cells and downregulated the expression of VEGF and matrix metalloproteinase 2 (MMP2). Additionally, Tang et al. (J Int Med Res (2012) 40(1):85-94) demonstrated that exposure to moderate AC electric fields reduced VEGF expression in mouse melanoma cell lines and mouse tumor models. Furthermore, Kim et al. (Oncotarget (2016) 7:65125-65136) demonstrated that TT fields inhibited the migration, invasion, and angiogenesis of glioblastoma cells, arguing that TT fields are a promising anti-invasive and anti-angiogenic therapeutic strategy for use in GBM patients.

[0007] Despite the availability of oncology-based therapies, glioblastoma multiforme (GBM) remains the most common and aggressive primary malignant tumor of the central nervous system in adults. The current standard of care for recurrent GBM is bevacizumab (AVASTIN®, Genentech, 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 may be effective for GBM (Shiyu et al., Biomedicine & Pharmacotherapy (2021) 141:111810). However, patients inevitably develop resistance to bevacizumab, and overall survival often does not improve significantly.

[0008] In Ansstas et al. (Case Rep Neurol (2016) 8:1-9), a "pulse-dose" approach to bevacizumab administration was combined with TT field therapy; patients with recurrent GBM discontinued bevacizumab, were subsequently treated with TT field therapy alone, and then, upon the onset of symptoms or radiographic evidence of progression, bevacizumab was re-administered in a "pulse-dose" fashion. The results of this study support the use of TT field therapy in combination with pulse-dose bevacizumab as an option for patients with refractory GBM. In Fallah et al. (J Clin Oncology (2020) 38(15_suppl):2537), the combination of bevacizumab and TT field therapy was shown to be safe, feasible, and clinically effective in patients with recurrent GBM.

[0009] Jo et al. (Int J Mol Sci (2018) 19(11):3684) studied the effect of sorafenib on the antitumor and antiangiogenic activity of TT cells in glioblastoma cells and found that the combination therapy inhibited tumor cell motility, invasiveness, and angiogenesis.

[0010] 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) investigated the addition of TT field therapy to the treatment of hepatocellular carcinoma (HCC, an aggressive liver cancer and one of the leading causes of cancer-related deaths worldwide) with sorafenib, a multikinase inhibitor and the primary first-line treatment for advanced HCC. The TT field / sorafenib combination therapy demonstrated improved response rates compared with conventional controls in patients with advanced HCC. [Brief explanation of the drawings]

[0011] [Figure 1] Included are representative images of tumor-bearing ovaries from an orthotopic ovarian cancer mouse model after 10 days of treatment with a 200 kHz TT field. [Figure 2] We show that serum circulating VEGF expression is upregulated after 10 days of treatment with a 200 kHz TT field in an ovarian cancer mouse model. [Figure 3] We show that serum circulating VEGF expression is upregulated after 14 days of treatment with a 150 kHz TT field in a lung cancer mouse model. [Figure 4] Figure 1 shows that intratumoral CD31 expression is upregulated after 10 days of treatment with a 200 kHz TT field in an ovarian cancer mouse model. [Figure 5] The experimental design of this lung cancer efficacy study in the LL2 orthotopic lung cancer model using TT field, DC101, and docetaxel is shown. [Figure 6] FIG. 1 shows the results of an example showing the effect of combining a TT field with DC101 and docetaxel. [Figure 7] FIG. 1 shows the results of an example showing the effect of combining a TT field with DC101 and docetaxel. DETAILED DESCRIPTION OF THE INVENTION

[0012] Before describing at least one embodiment of the inventive concept(s) in detail using illustrative language and results, it should be understood that application of the inventive concept(s) is not limited to the details of construction and the arrangement of components set forth in the following description. The inventive concept(s) may be embodied in other embodiments or may be practiced or carried out in various ways. Therefore, the words used herein are intended to be accorded the broadest possible scope and meaning. Also, the embodiments are intended to be illustrative, not exhaustive. It should also be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting.

[0013] Unless otherwise defined herein, scientific and technical terms used in connection with the concept(s) of the present invention shall have the meanings commonly understood by those of ordinary skill in the art. Furthermore, unless otherwise required by context, specific terms include pluralities and plural terms include the specific terms. The foregoing techniques and procedures are generally carried out according to conventional methods well known in the art and as described in the various general and more specific references cited and described throughout the specification. The nomenclature used in connection with analytical chemistry, synthetic organic chemistry, medicinal chemistry, and pharmaceutical chemistry described herein, as well as the experimental procedures and techniques thereof, are well known and commonly used in the art. Standard techniques are used for chemical syntheses and chemical analyses.

[0014] All patents, published patent applications, and non-patent publications mentioned in this specification are indicative of the level of skill of those skilled in the art to which the presently disclosed inventive concept(s) pertain. All patents, published patent applications, and non-patent publications referenced in any part of this application are 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.

[0015] All of the compositions, assemblies, systems, kits, and / or methods disclosed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions, assemblies, systems, kits, and methods of the inventive concept(s) have been described with reference to specific embodiments, it will be apparent to those skilled in the art that variations can be applied to the compositions and / or methods, and to the steps or sequence of steps of the methods described herein without departing from the concept(s), spirit, and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope, and concept of the inventive concept(s) as defined by the appended claims.

[0016] As used in accordance with the present disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0017] The use of the terms "a" or "an" in the claims and / or specification when used in conjunction with the term "comprising" may mean "one," but is also consistent with the meaning of "one or more," "at least one," or "one or more." Thus, unless the context clearly dictates otherwise, the terms "a," "an," and "the" include plural references. Thus, for example, "a compound" can refer to one or more compounds, two or more compounds, three or more compounds, four or more compounds, or more compounds. The term "plurality" refers to two or more.

[0018] Use of the term "at least one" is understood to include 1, as well as quantities greater than 1, including, but not limited to, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100, etc. The term "at least one" may extend to 100 or 1000 or more, depending on the term to which it is attached, and the quantity 100 / 1000 is not considered limiting, as higher limits may also be satisfactory. Additionally, use of the term "at least one of X, Y, Z" is understood to include X alone, Y alone, Z alone, and any combination of X, Y, and Z. Use of ordinal terms (e.g., "first," "second," "third," "fourth," etc.) is intended only to distinguish between two or more items and does not imply any order of superiority or importance of one item over another, or any additional order.

[0019] The use of the term "or" in the claims is used to mean an inclusive "and / or" unless explicitly stated to refer only to alternatives or unless the alternatives are mutually exclusive. For example, a condition "A or B" can be satisfied by any of the following: A being true (or existing) and B being false (or not existing), A being false (or existing) and B being true (or existing), and both A and B being true (or existing).

[0020] As used herein, references to "one embodiment," "embodiment," "some embodiments," "an example," "for example," or "one example" mean that a particular element, feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment. For example, the appearances of the phrases "in some embodiments" or "in one example" in various places in the specification do not necessarily all refer to the same embodiment. Moreover, all references to one or more embodiments or examples should not be construed as limiting the claims.

[0021] Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for a composition / apparatus / device, the method used to determine the value, or the variation that exists between study subjects. For example, and without limitation, when the term "about" is used, the specified value may vary by plus or minus 20 percent, 15 percent, 12 percent, 11 percent, 10 percent, 9 percent, 8 percent, 7 percent, 6 percent, 5 percent, 4 percent, 3 percent, 2 percent, or 1 percent from the specified value, as appropriate for performing the disclosed methods and as understood by one of ordinary skill in the art.

[0022] As used in this specification and claims, the terms "comprising" (and any form of "comprise" or "comprises"), "having" (and any form of "have" or "has"), "including" (and any form of "includes" or "include"), and "containing" (and any form of "contains" or "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0023] As used herein, the term "or combinations thereof" refers to all permutations and combinations of the listed items preceding 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, where order is important in the particular context, also at least one of BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, combinations including repeats of one or more items or terms are expressly included, such as BB, AAA, AAB, BBC, AAABCCCC, CBBAAA, CABABB, etc. One of ordinary skill in the art will understand that there is typically no limit to the number of items or terms that may be combined unless otherwise apparent from the context.

[0024] As used herein, the term "substantially" means that the subsequently described event or circumstance occurs entirely, or that the subsequently described event or circumstance occurs to a large extent or degree. For example, when relating to a particular event or circumstance, the term "substantially" means that the subsequently described event or circumstance occurs at least 80% of the time, at least 85% of the time, 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, that two items are very close to each other but not 100% adjacent, or that a portion of one of two items is very close to the other item but not 100% adjacent to the other item.

[0025] The term "pharmaceutically acceptable" refers to compounds and compositions that are suitable for administration to humans and / or animals without undue side effects, such as toxicity, irritation, and / or allergic response, commensurate with a reasonable benefit / risk ratio.

[0026] As used herein, the term "patient" or "subject" includes human and veterinary subjects. For purposes of treatment, "mammal" refers to animals classified as mammals, including, but not limited to, humans, domestic and farm animals, non-human primates, and other animals with mammary tissue.

[0027] The term "treatment" refers to both therapeutic and prophylactic measures. Those in need of treatment include, but are not limited to, those who already suffer from a particular condition / disease / infection, as well as those who are at risk of contracting a particular condition / disease / infection (such as those who require prophylactic measures). The term "treating" refers to the administration of an agent / substance / method to a patient for therapeutic and / or prophylactic purposes.

[0028] 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 effect.

[0029] Administration of a therapeutically or prophylactically effective amount is intended to provide a therapeutic benefit in the treatment, prevention, and / or management of a disease, condition, and / or infection. The specific amount that is therapeutically effective can be readily determined by an ordinary medical practitioner and can vary depending on factors known in the art, such as, but not limited to, the type of condition / disease / infection, the patient's medical history and age, the stage of the condition / disease / infection, and the co-administration of other drugs.

[0030] The term "effective amount," when used in accordance with the concept(s) of the present invention, refers to an amount of a biologically active molecule or its conjugate or derivative, or an amount of a treatment protocol (e.g., an alternating electric field), sufficient to exert a detectable therapeutic effect without undue side effects (such as, but not limited to, toxicity, irritation, allergic reaction, etc.) commensurate with a reasonable benefit / risk ratio. A therapeutic effect may include, for example, but is not limited to, preventing, inhibiting, or reducing the occurrence of at least one condition, disease, and / or infection. The effective amount for a subject will vary depending on the type of subject, the subject's size and health, the nature and severity of the condition / disease / infection being treated, the method of administration, the duration of treatment, the nature of concomitant therapy (if any), the particular formulation used, and the like. Thus, an exact effective amount cannot be specified in advance. However, an effective amount for a particular situation can be determined by one of ordinary skill in the art using routine experimentation based on the information provided herein.

[0031] As used herein, the term "concomitant therapy" is used interchangeably with the terms "combination therapy" and "adjunctive therapy" and is understood to mean that a patient in need thereof is treated with or administered another agent for a condition / disease / infection in conjunction with the treatment of the present disclosure. This concomitant therapy may be sequential therapy, where the patient is treated first with one therapeutic protocol / pharmaceutical composition and then with another therapeutic protocol / pharmaceutical composition, or where two therapeutic protocols / pharmaceutical compositions are administered simultaneously.

[0032] As used herein, the terms "administration" and "administering" are understood to encompass 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 to encompass both local and systemic application. In addition, the compositions of the present disclosure (and / or methods of administration thereof) may be designed to provide delayed-, controlled-, or sustained-release using formulation techniques well known in the art.

[0033] Turning now to the concept(s) of the present invention, a combination therapy for cancer is disclosed herein. The combination therapy includes the use of an alternating current electric field (e.g., a TT field) in combination with at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (i.e., an anti-VEGFR-2 antibody) and inhibits the action of VEGFR-2 (e.g., by inhibiting the interaction between VEGF and VEGFR-2, but is not limited to this). The combination of the alternating current electric field (e.g., a TT field) and the anti-VEGFR-2 antibody produces a synergistic effect in the treatment of cancer.

[0034] Certain non-limiting embodiments of the present disclosure relate to a method for reducing the viability of cancer cells, the method comprising: (1) administering to the cancer cells at least one composition, the at least one composition comprising at least one antibody that specifically binds to vascular endothelial growth factor receptor 2, which specifically interacts with VEGFR-2, and inhibits the interaction between VEGF and VEGFR-2; and (2) applying an alternating electric field to the cancer cells for a period of time.

[0035] Certain additional non-limiting embodiments of the present disclosure relate to a method of treating cancer in a subject, the method comprising: (1) administering to the subject at least one composition comprising at least one anti-VEGFR-2 antibody that specifically interacts with VEGFR-2 and inhibits the interaction between VEGF and VEGFR-2; and (2) applying an alternating electric field to a target region of the subject.

[0036] Certain additional non-limiting embodiments of the present disclosure relate to a method for reducing the volume of a tumor present in a living body, the tumor comprising a plurality of cancer cells, comprising: (1) administering to a subject at least one composition comprising at least one anti-VEGFR-2 antibody that specifically interacts with VEGFR-2 and inhibits the interaction between VEGF and VEGFR-2; and (2) applying an alternating electric field to a target region of the subject.

[0037] Certain additional non-limiting embodiments of the present disclosure relate to a method for preventing an increase in tumor volume, the tumor being present in a living organism and comprising a plurality of cancer cells, the method comprising: (1) administering to a subject at least one composition comprising at least one anti-VEGFR-2 antibody that specifically interacts with VEGFR-2 and inhibits the interaction between VEGF and VEGFR-2; and (2) applying an alternating electric field to a target region of the subject.

[0038] Steps (1) and (2) of any of the methods of the present disclosure can be performed simultaneously, sequentially, and in particular, substantially simultaneously, or wholly or partially sequentially. When the steps are performed wholly or partially sequentially, at least one composition comprising at least one anti-VEGFR-2 antibody can be administered before or after application of the alternating electric field is initiated.

[0039] The methods of the present disclosure can be utilized to treat any type of cancer cell / cancer / tumor that responds to treatment with an alternating electric field (e.g., a TT field) and / or an anti-VEGFR-2 antibody. Non-limiting examples of cancer cells / cancers / tumors that can be treated according to the present disclosure 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.

[0040] Any type of conductive or non-conductive electrode(s) and / or transducer array(s) known in the art or otherwise discussed herein that can be used to generate an alternating electric field can be utilized to generate an alternating electric field in accordance with the methods of the present disclosure. Non-limiting examples of electrodes and transducer arrays that can be utilized to generate alternating electric fields in accordance with the present disclosure include, but are not limited to, those that function as part of the TT field systems described, for example, in U.S. Pat. Nos. 7,016,725, 7,089,054, 7,333,852, 7,565,205, 8,244,345, 8,715,203, 8,764,675, 10,188,851, and 10,441,776, and U.S. Pat. Nos. 2018 / 0160933, 2019 / 0117956, 2019 / 0307781, and 2019 / 0308016.

[0041] The AC electric field may be generated at any frequency according to the present disclosure. For example (but not limited to), the AC electric field may be generated at any frequency, including 50 kHz, 75 kHz, 100 kHz, 125 kHz, 150 kHz, 175 kHz, 200 kHz, 225 kHz, 250 kHz, 275 kHz, 300 kHz, 325 kHz, 350 kHz, 375 kHz, 400 kHz, 425 kHz, 450 kHz, 475 kHz, 500 kHz, 550 kHz, 600 kHz, 650 kHz, 700 kHz, 750 kHz, 800 kHz, 850 kHz, 900 kHz, 950 kHz, 1 MHz, 2 MHz, 3 MHz, 4 MHz, The frequency may be 5 MHz, 6 MHz, 7 MHz, 8 MHz, 9 MHz, 10 MHz, etc., as well as a range formed from 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 of about 150 kHz to about 300 kHz, etc.), and a range combining two integers between two of the above values ​​(e.g., a range of about 122 kHz to about 313 kHz, a range of about 78 kHz to about 298 kHz, etc.).

[0042] In certain (but non-limiting) embodiments, the alternating electric field may be applied at two or more different frequencies, where two or more frequencies are present, each frequency being selected from any of the values ​​above, or a range formed from any of the values ​​above, or a range combining two integers between any two of the values ​​above.

[0043] The alternating electric field can have any electric field strength within the subject / cancer cells as long as it is capable of functioning in accordance with the present disclosure. For example (but not limited to), the alternating electric field may be 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 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, etc., as well as ranges formed from any of the above values ​​(e.g., 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.), and ranges formed by combining two integers between two of the above values ​​(e.g., 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.).

[0044] The AC electric field may be applied in a single direction between one pair of arrays, or in two directions / channels (e.g., front-to-back and side-to-side) between two pairs of arrays. For example, certain TT field devices (OPTUNE® system (Novocure, St. Helier, Jersey)) operate in two directions to increase the likelihood that dividing cells will align with the electric field, allowing the electric field to exert its desired antimitotic effect. However, it is understood that the scope of the present invention also includes the application of an AC electric field in a single direction. As used herein, the term "AC electric field" is understood to include not only application in a single direction / channel but also application in two directions / channels. Furthermore, as used herein, the term "AC electric field" is understood to include not only application of a single AC electric field but also application of multiple AC electric fields sequentially over a period of time.

[0045] The AC electric field can be applied for any duration or cumulative time sufficient to achieve a reduction in cancer cell viability and / or a reduction in tumor volume (and / or prevention of tumor volume increase). The period during which the AC electric field is applied includes both continuous and cumulative periods. That is, the period during which the AC electric field is applied includes not only a single session (i.e., continuous application) but also multiple sessions with short breaks between sessions (i.e., continuous application over a cumulative period). For example, subjects may be allowed to take breaks during treatment with the AC electric field device, and are expected to have the device in place and operational on their body for only about 60%, at least about 70%, or at least about 80% of the entire treatment period (e.g., 1 day, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, etc.).

[0046] For example, but not by way of limitation, the AC electric field may 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 78 hours, about 81 hours, about 84 hours, about 87 hours, about 90 hours, or the like. The radiation exposure may be applied for a continuous or cumulative period of time such as about 1 hour, 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., as well as a range formed from any of the above values ​​(e.g., a range of about 1 hour to about 6 months, a range of about 24 hours to about 72 hours, etc.), and a range formed by combining two integers between any two of the above values ​​(e.g., a range of about 14 hours to about 68 hours, etc.).

[0047] In a particular (but non-limiting) embodiment, the alternating electric field is applied for a period of at least about 24 hours.

[0048] Any anti-VEGFR-2 antibody known in the art or otherwise discussed herein can be used in accordance with the present disclosure, as long as the agent specifically interacts with VEGFR-2 and can inhibit the interaction between VEGF and VEGFR-2. Furthermore, the anti-VEGFR-2 antibody can be a polyclonal or monoclonal antibody. Non-limiting examples of monoclonal anti-VEGFR-2 antibodies that can be used in accordance with the present disclosure include ramucirumab, DC101, etc., and any combination thereof. Both ramucirumab and DC101 (and other anti-VEGFR-2 monoclonal antibodies) are known in the art and are commercially available from multiple sources, and therefore further description thereof is not considered necessary.

[0049] Compositions comprising at least one anti-VEGFR-2 antibody can be provided in any formulation known in the art or otherwise discussed herein. For example (but not limited to), the composition can further comprise trifluridine / uracil (FTD / TPI), which is routinely utilized in combination with monoclonal antibody DC101.

[0050] In certain (but non-limiting) embodiments, compositions containing at least one anti-VEGFR-2 antibody include one or more pharmaceutically acceptable carriers (therefore, the compositions are also referred to as "pharmaceutical compositions"). Non-limiting examples of suitable pharmaceutically acceptable carriers include water, saline, dextrose solution, fructose or mannitol, calcium carbonate, cellulose, ethanol, oils of animal, vegetable, or synthetic origin, carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low-molecular-weight proteins, detergents, liposome carriers, sodium chloride, buffer solutions such as saline and phosphate-buffered saline, and / or other substances, diluents, excipients such as polyethylene glycol (PEG), or any combination thereof that are physiologically acceptable and / or safe to use. Suitable pharmaceutically acceptable carriers for pharmaceutical formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 23rd ed. (2020).

[0051] In certain (but non-limiting) embodiments, compositions comprising at least one anti-VEGFR-2 antibody may further contain one or more additional activating agents. Various active drugs for use in combination with AC electric fields or various VEGF / VEGFR inhibitors are known in the art, and certain combination therapies have been approved by the FDA or are currently in clinical trials. Non-limiting examples of therapeutic agents that can be used in combination with anti-VEGFR-2 antibodies in accordance with the present disclosure include, but are not limited to, anti-PD-1 therapeutic agents such as pembrolizumab (Keytruda), tislelizumab, nivolumab, and cemiplimab; anti-PD-L1 therapeutic agents such as atezolizumab, avelumab, and durvalumab; lenvatinib, paclitaxel, docetaxel, ifosamide, etoposide (bepecid), gemcitabine, lomustine, nab-paclitaxel, temozolomide, and anti-CTLA-4 therapeutic agents; aromatase inhibitors such as letrozole (but not limited to), biologics such as monoclonal antibodies (for example, but not limited to, denosumab and pembrolizumab); and any combination thereof.

[0052] In addition, any composition of the present disclosure may contain other drugs that allow the composition to be administered by a specific administration route. For example, but not limited to, the composition may be formulated for administration by oral, topical, transdermal, parenteral, subcutaneous, intranasal, mucosal, intramuscular, intraperitoneal, intravitreal, and / or intravenous routes. Depending on the administration route, the composition may also contain one or more additional components (e.g., anti-VEGFR-2 antibody(ies) and / or additional therapeutic agent(s)) in addition to the activator. Examples of additional secondary compounds that may be present may include, but are not limited to, fillers, salts, buffers, preservatives, stabilizers, solubilizers, wetting agents, emulsifiers, dispersants, and other substances well known in the art.

[0053] In a specific, but non-limiting, embodiment, at least one composition comprising at least one anti-VEGFR-2 antibody is orally administered to a subject.

[0054] At least one composition comprising at least one anti-VEGFR-2 antibody may be administered before or after application of an alternating current electric field is initiated. In certain (but non-limiting) embodiments, at least one composition comprising at least one anti-VEGFR-2 antibody may be administered after application of an alternating current electric field is initiated. In particular (but not limited to), at least one composition comprising at least one anti-VEGFR-2 antibody may be administered during application of the alternating current electric field (e.g., before the period during which the alternating current electric field is applied has elapsed) and / or after application of the alternating current electric field has elapsed.

[0055] For example, but not limited to, at least one composition comprising at least one anti-VEGFR-2 antibody may be administered for 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, or about 66 hours after the start of application of an alternating current electric field. The administration may be performed after 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., as well as a range formed by any of the above values ​​(e.g., a range of about 24 hours to about 96 hours), and a range formed by combining two integers between two of the above values ​​(e.g., a range of about 14 hours to about 94 hours). In a specific (but non-limiting) embodiment, at least one composition comprising at least one anti-VEGFR-2 antibody is administered at least about 24 hours after the start of application of the alternating current electric field.

[0056] In other non-limiting examples, the at least one composition comprising at least one anti-VEGFR-2 antibody can be administered after the period of time during which the alternating current electric field has been applied, where the at least one composition comprising at least one anti-VEGFR-2 antibody 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 period of time during which the alternating current electric field has been applied.

[0057] In a specific, but non-limiting, embodiment, at least one composition comprising at least one anti-VEGFR-2 antibody is administered within about 96 hours after the period has elapsed.

[0058] Compositions comprising at least one anti-VEGFR-2 antibody can be administered to cancer cells / subjects at any concentration that provides a therapeutically effective concentration of the at least one anti-VEGFR-2 antibody. In certain non-limiting embodiments, application of an AC electric field reduces the amount of anti-VEGFR-2 antibody required for therapeutic efficacy compared to the typical therapeutically effective amount administered in the absence of an AC electric field. For example, but not by way of limitation, the therapeutically effective concentration of an anti-VEGFR-2 antibody can be reduced by at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more relative to an amount of anti-VEGFR-2 antibody known to be therapeutically effective in the absence of an AC electric field. In certain (but non-limiting) embodiments, the therapeutically effective concentration of an anti-VEGFR-2 antibody is reduced by at least about 50% compared to an amount of anti-VEGFR-2 antibody known to be therapeutically effective in the absence of an AC electric field.

[0059] In certain (but non-limiting) embodiments, the therapeutically effective concentration of an anti-VEGFR-2 antibody utilized in accordance with the present disclosure may 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, It can be about 55 nM, about 60 nM, about 65 nM, about 70 nM, about 75 nM, about 80 nM, about 85 nM, about 90 nM, about 95 nM, about 100 nM, about 110 nM, about 120 nM, about 130 nM, about 140 nM, about 150 nM, etc., as well as a range formed from any of the above values ​​(e.g., a range of about 12.5 nM to about 100 nM), and a range formed by combining two integers between two of the above values ​​(e.g., a range of about 17 nM to about 83 nM).

[0060] In certain (but non-limiting) embodiments, the therapeutically effective concentration of an anti-VEGFR-2 antibody is from about 12.5 nM to about 100 nM.

[0061] In certain (but non-limiting) embodiments, a therapeutically effective concentration of an anti-VEGFR-2 antibody utilized in accordance with the present disclosure can be, for example (but not limited to), about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, about 20 mg / kg, etc., as well as a range formed from any of the above values ​​(e.g., a range of about 1 mg / kg to about 20 mg / kg).

[0062] In certain (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 (3) discontinuing the application of the AC electric field (e.g., but not by way of limitation, to allow the cells / tissues to recover). Additionally, either steps (1) and / or (2) may be repeated one or more times.

[0063] In certain (but non-limiting) embodiments, the anti-VEGFR-2 antibody may be administered by any dosing regimen known in the art. For example, but not limited to, the anti-VEGFR-2 antibody may be administered in a single dose or multiple doses over a defined treatment period. For example (but not limited to), a therapeutically effective concentration of the anti-VEGFR-2 antibody may be administered approximately once every 4 hours, approximately once every 8 hours, approximately once every 12 hours, approximately once every day, approximately once every 2 days, approximately once every 3 days, approximately once a week, approximately twice a week, approximately three times a week, approximately once every 2 weeks, approximately once every 3 weeks, approximately once a month, etc., as well as ranges formed by any of the above values ​​(e.g., approximately once every 4 to 8 hours, approximately once a week to approximately once a month, etc.).

[0064] In certain (but non-limiting) embodiments, the method includes simultaneous treatment with two or more compositions. Thus, the method may include the additional step of (4) administering at least a second composition to the cancer cells / subject. In certain (but non-limiting) embodiments, the at least second composition may contain any one or more of the active agents disclosed or otherwise discussed herein for use with the small molecule antiangiogenic agent(s).

[0065] Various substances and therapies used in combination with VEGF / VEGFR inhibitors are known in the art, and certain combination therapies have been approved by the FDA or are currently in clinical trials. Non-limiting examples of therapeutic agents that can be used in combination with anti-VEGFR-2 antibodies in simultaneous (sequential) therapy in accordance with the present disclosure include, but are not limited to, anti-PD-1 therapeutic agents such as pembrolizumab (Keytruda), tislelizumab, nivolumab, and cemiplimab; anti-PD-L1 therapeutic agents such as atezolizumab, avelumab, and durvalumab; lenvatinib, paclitaxel, docetaxel, ifosamide, etoposide (bepecid), gemcitabine, lomustine, nab-paclitaxel, temozolomide, rifametinib ... anti-CTLA-4 therapeutic agents; aromatase inhibitors such as letrozole (but not limited to), biologics such as monoclonal antibodies (for example, but not limited to, denosumab and pembrolizumab); and any combination thereof.

[0066] If present, step (4) may be performed substantially simultaneously with the administration of the first composition in step (1) or sequentially in whole or in part, thereby administering two separate compositions simultaneously or sequentially in whole or in part. In addition, the two compositions administered in steps (1) and (4) may be administered by the same route (e.g., both orally), or the two compositions may be administered by different routes (e.g., one composition administered orally and another composition administered intravenously).

[0067] If present, the optional additional administration step (4) may be carried out in the same manner(s) and time frame(s) as described above for the first composition, before or after application of the alternating electric field is initiated, during application of the alternating electric field, and / or after application of the alternating electric field has elapsed.

[0068] That is, for example (but not limited to), the second composition may remain in a state where it remains ... , 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, and similarly, a range formed from any of the above values ​​(e.g., about 24 hours to about 96 hours), and a range formed by combining two integers between two of the above values ​​(e.g., about 14 hours to about 94 hours). In a specific (but non-limiting) embodiment, the second composition is administered at least about 24 hours after application of the alternating electric field has begun.

[0069] In other non-limiting examples, the second composition can be administered after the period of time during which the alternating electric field has been applied, in which case the second composition 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 period of time during which the alternating electric field has been applied. In certain (but non-limiting) embodiments, the second composition is administered within about 96 hours after the period of time has elapsed.

[0070] Additionally, for example (but not limited to), the second composition may be administered within 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 45 hours, about 48 hours after the first agent is administered. , 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., as well as a range formed from any of the above values ​​(e.g., a range of about 24 hours to about 96 hours), and a range formed by combining two integers between the above two values ​​(e.g., a range of about 14 hours to about 94 hours). In a specific (but non-limiting) embodiment, the second composition is administered at least about 12 hours after the first agent is administered.

[0071] In certain (but non-limiting) embodiments, the method may further comprise the step of (5) administering at least one additional therapy to the cells / subject. Any therapy known in the art or contemplated herein for use in combination with alternating electric fields (e.g., TT fields) and / or anti-VEGFR-2 antibody therapy may be utilized in accordance with the methods of the present disclosure. Non-limiting examples of additional therapies that may be utilized include radiation therapy, photodynamic therapy, transarterial chemoembolization (TACE), or combinations thereof.

[0072] Steps (1) and (2) and any of optional steps (3), (4), and (5) may be repeated one or more times. Each step may be repeated as many times as necessary. When step (2) is repeated, the transducer array may be positioned slightly differently from its original position on the subject, and this repositioning of the array may further facilitate tumor / cancer treatment. Additionally, step (1) and optional steps (4) and (5) (if present) of administering the composition(s) / additional therapy(ies) may be repeated various times and at various intervals to conform to known and / or commonly accepted dosage / treatment regimens for the compositions / therapies.

[0073] The use of ordinal numbers for any steps is for illustrative purposes only, and the methods of the present disclosure may include any one or more of steps (3), (4), and (5) alone or in combination with each other, i.e., the methods of the present disclosure include performing step (3) if step (4) or (5) is not present, performing step (4) if step (3) or (5) is not present, and performing step (5) if steps (3) and (4) are not present. In other words, the scope of the method disclosed herein includes performing steps (1)-(2) (repeat each step as many times as necessary), performing steps (1)-(3) (repeat one or more of steps (1)-(3) as many times as necessary), performing steps (1)-(2) and (4) (repeat one or more of steps (1)-(2) and (4) as many times as necessary), performing steps (1)-(2) and (5) (repeat one or more of steps (1)-(2) and (5) as many times as necessary), and performing steps ( This includes performing steps (1) to (4) (repeat one or more of steps (1) to (4) as many times as necessary), performing steps (1) to (3) and (5) (repeat one or more of steps (1) to (3) and (5) as many times as necessary), performing steps (1) to (2) and (4) to (5) (repeat one or more of steps (1) to (2) and (4) to (5) as many times as necessary), and performing all of steps (1) to (5) (further repeat one or more of steps (1) to (5) as many times as necessary).

[0074] While the above explicitly describes the use of a combination therapy of two agents, it is understood that the scope of the present disclosure also includes combination therapy of three or more compositions. Accordingly, the method can include one or more additional steps (similar to steps (1) and (4)) of administering an additional composition to the subject. The additional agent administered in this method can be selected from any of the agents disclosed herein or contemplated for use in combination with at least one anti-VEGFR-2 antibody (as disclosed herein above with respect to optional step (4)). Furthermore, the administration of the additional agent can be substantially simultaneous with or completely or partially sequential to the administration of the first composition / agent and the second composition / agent, and in the same manner(s) and time frame(s) as described above for the first composition / agent and the second composition / agent.

[0075] Certain non-limiting embodiments of the present disclosure include any of the alternating electric field (e.g., TT field) generating systems disclosed herein or otherwise discussed (e.g., U.S. Pat. Nos. 7,016,725, 7,089,054, 7,333,852, 7,565,205, 8,244,345, 8,715,203, 8,764,675, 10,188,851, and 10,441,776, and U.S. Patent Application No. 2018 / 016 The present invention relates to kits that include a composition comprising one or more transducer arrays and / or one or more hydrogel compositions (such as, but not limited to, those disclosed in US Pat. Nos. 2019 / 0933, 2019 / 0117956, 2019 / 0307781, and 2019 / 0308016) in combination with at least one of any of the compositions comprising at least one anti-VEGFR-2 antibody disclosed herein or otherwise discussed. The kits may optionally further contain one or more of any of the compositions disclosed herein or otherwise discussed (such as any one or more compositions comprising at least one additional activating agent). The kits may optionally further include one or more devices (or one or more components of a device) used in one or more additional treatment steps.

[0076] In certain (non-limiting) embodiments, the kit may further include instructions for carrying out any of the methods disclosed herein or otherwise discussed. For example (but not limited to), the kit may include instructions for applying one or more components of an alternating current electric field (e.g., TT field) generating device to a patient's skin, instructions for applying an alternating current electric field to a patient, instructions regarding when and how to administer a composition comprising at least one anti-VEGFR-2 antibody, and optionally, how to administer one or more optional additional compositions, and / or instructions regarding when to activate and deactivate the alternating current electric field in connection with the administration of a composition comprising at least one anti-VEGFR-2 antibody, and / or the administration of one or more optional compositions, and / or a treatment step.

[0077] In addition to the components detailed above, the kits may further contain other component(s) / reagent(s) for carrying out any of the specific methods described herein or otherwise discussed. For example (but not limited to), the kits may further include: (i) components for preparing the skin prior to discarding the hydrogel composition and / or transducer array on the skin (e.g., razors, cleaning compositions, wipes / towels, etc.); (ii) components for removing the gel / transducer array(s); (iii) components for cleaning the skin after removing the gel / transducer array(s); and / or (iv) other components used in the system (i.e., conductive materials, non-conductive materials, soothing gels or creams, bandages, etc.). The nature of these additional component(s) / reagent(s) will depend on the particular treatment format, and their identification is within the skill of those of ordinary skill in the art, and therefore further description is not believed necessary. Additionally, the components / reagents included in the kit may each be contained in separate containers / compartments, 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.

[0078] The kit can be disposed in any packaging that enables the components contained therein to function according to the present disclosure. In certain non-limiting embodiments, the kit further comprises a sealed package into which the components are disposed. In certain (but non-limiting) embodiments, the sealed package is substantially impermeable to air and / or substantially impermeable to light.

[0079] Additionally, the kit may further include a set of written instructions describing how to use one or more components of the kit. Kits of this nature may be used in any of the methods described herein or otherwise discussed.

[0080] 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.

[0081] Certain non-limiting embodiments of the present disclosure include any of the AC electric field generating systems disclosed herein or otherwise discussed (e.g., U.S. Pat. Nos. 7,016,725, 7,089,054, 7,333,852, 7,565,205, 8,244,345, 8,715,203, 8,764,675, 10,188,851, and 10,441,776, and U.S. Patent Application No. 2018 / 0160933). The present invention relates to kits that include a system comprising one or more transducer arrays and / or one or more hydrogel compositions (such as, but not limited to, those disclosed in US Pat. Nos. 2019 / 0117956, 2019 / 0307781, and 2019 / 0308016) in combination with at least one of any of the compositions comprising at least one anti-VEGFR-2 antibody disclosed herein or otherwise discussed. The system may optionally further include one or more of any of the compositions disclosed herein or otherwise discussed. 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. Example

[0082] The following examples are provided. However, it should be understood that the present disclosure is not limited in its application to the specific experiments, results, and experimental procedures disclosed herein below. Rather, this example is provided merely as one of various embodiments and is intended to be illustrative rather than exhaustive. Example 1

[0083] While the prior art referenced in the Background section above has shown that TT fields have anti-angiogenic activity, this example demonstrates that TT field treatment increases VEGF expression and indeed increases the formation of new blood vessels.

[0084] Mice were injected with MOSE-L-FFL (day 0) to create an orthotopic ovarian cancer mouse model, which was confirmed by IVIS on day 15. Starting on day 16, mice were treated with a 200 kHz TT field (or heat control) for 10 days. As seen in Figure 1, increased vascularity was observed in the tumor-filled ovaries of TT field-treated mice compared to untreated controls (heat-treated mice).

[0085] Mice from this in vivo ovarian cancer model were sacrificed on day 26, and blood samples were collected and collected in designated serum tubes. They were centrifuged at 1000 g for 15 minutes and stored at -20°C. The serum was incubated on a cytokine array membrane, and as shown in Figure 2, increased VEGF secretion was observed after TT field treatment compared to heat-treated mice.

[0086] Next, mice with rectal lung cancer were generated by injecting LLC-2 (day 0) and confirming the cancer on day 7. Starting on day 7, the mice were treated with a 150 kHz TT field for 14 days. On day 21, the mice were sacrificed, and blood samples were collected in designated serum tubes, centrifuged at 1000 g for 15 minutes, and stored at -20°C. The serum was incubated on a cytokine array membrane, and as shown in Figure 3, an increase in secreted VEGF was observed after the TT field compared to heat-treated mice.

[0087] Next, mice from the same in vivo ovarian cancer model as in Figures 1-2 were sacrificed on day 26, and tumor samples were collected for histological analysis. Tumor tissue was stained by immunohistochemical staining for CD31 (a marker for endothelial cells and blood vessels), and images were quantified. CD31 staining was indicated by brown (DAB staining). As shown in Figure 4, the microvascular density of tumors treated with TT field was significantly higher than that of control tumors treated with heat.

[0088] Thus, in contrast to the current literature suggesting that TT field treatment has an anti-angiogenic effect, this example demonstrates that application of a TT field actually has a pro-angiogenic effect, resulting in increased VEGF expression and the formation of new vasculature to supply the tumor. Thus, administration of an anti-VEGFR-2 antibody counteracted the pro-angiogenic effect of TT field administration in the combination therapy. Example 2

[0089] In this example, the efficacy of a TT field in combination with DC101, a murine version of ramucirumab, and the chemotherapy drug docetaxel was studied in a lung cancer model. Figure 5 shows the experimental design for this lung cancer efficacy study in the LL2 orthotopic lung cancer model using a TT field, DC101, and docetaxel. C57Bl / 6 mice were injected with LLC-2 (day 0) and confirmed on day 6 to generate mice with orthotopic lung cancer. Starting on day 7, mice were treated with a 150 kHz TT field or heat sham (control) for 7 days. Mice were also injected with 5 mg / kg DC101 on days 7, 10, and 13, and 3 mg / kg docetaxel on days 8 and 12. On day 15, mice were sacrificed, and tumors were excised and examined.

[0090] As can be seen from Figures 6-7, the combination of TT fields with DC101 and docetaxel had a significant effect on reducing tumor weight as well as tumor volume. Example 3

[0091] In this example, human subjects are treated with a TT field by applying an OPTUNE® device (Novocure Limited, St. Helier, Jersey) to the subject's skin with a pair of arrays positioned left-right (LR) and / or anterior-posterior (AP) of the tumor. Each subject is then chronically treated with a 150-200 kHz TT field. The device is worn at least approximately 80% of the time, with slight breaks between sessions to allow for slight adjustments to array placement and for cells and skin recovery.

[0092] Two weeks after application of the TT field, human subjects receive 10 mg / kg of ramucirumab once every three days for 14 days.

[0093] Following combination therapy treatment, the effect of the combination therapy on the tumor(s) is assessed. Non-Limiting Exemplary Embodiments of the Inventive Concept(s)

[0094] Exemplary Embodiment 1. A method for reducing the viability of cancer cells, the method comprising: (1) administering to the cancer cells at least one composition, wherein the at least one composition comprises at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (anti-VEGFR-2 antibody); and (2) applying an alternating current electric field to the cancer cells for a period of time.

[0095] Exemplary embodiment 2. The method of exemplary embodiment 1, wherein the alternating electric field is applied at a frequency in the range of about 50 kHz to about 10 MHz or about 50 kHz to about 1 MHz, the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells, and the alternating electric field is applied for at least about 24 hours.

[0096] Exemplary Embodiment 3. The method of exemplary embodiment 1 or 2, wherein said at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

[0097] Exemplary Embodiment 3a. The method of Exemplary Embodiment 3, wherein the at least one anti-VEGFR-2 antibody comprises DC101, and the composition further comprises trifluridine / tipiracil (FTD / TPI).

[0098] Exemplary Embodiment 3b. The method of exemplary embodiment 1 or 2, wherein said at least one anti-VEGFR-2 antibody is a polyclonal antibody.

[0099] Exemplary Embodiment 4. The method of any one of Exemplary Embodiments 1-3, wherein steps (1) and (2) are performed substantially simultaneously.

[0100] Exemplary Embodiment 5. The method of any one of Exemplary Embodiments 1-3, wherein steps (1) and (2) are performed sequentially, in whole or in part, and wherein the at least one composition is administered before the application of the alternating electric field begins.

[0101] Exemplary Embodiment 6. The method of any one of Exemplary Embodiments 1-3, wherein steps (1) and (2) are performed sequentially, in whole or in part, and the at least one composition is administered after the application of the alternating electric field begins.

[0102] Exemplary Embodiment 6a. The method of Exemplary Embodiment 6, wherein the at least one composition is administered before the period of time during which the alternating electric field is applied has elapsed.

[0103] Exemplary Embodiment 6b. The method of claim 6, wherein said at least one composition is administered after said period of time has elapsed.

[0104] Exemplary Embodiment 7. The method of any one of Exemplary Embodiments 1-6, wherein steps (1) and (2) are repeated one or more times.

[0105] Exemplary Embodiment 8. The method of any one of Exemplary Embodiments 1-7, 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.

[0106] Exemplary embodiment 9. A method of treating cancer in a subject, the method comprising: (1) administering to the subject at least one composition, wherein the at least one composition comprises an anti-VEGFR-2 antibody; and (2) applying an alternating electric field to a target area of ​​the subject.

[0107] Exemplary Embodiment 10 The method of Exemplary Embodiment 9, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

[0108] Exemplary Embodiment 10a. The method of exemplary embodiment 10, wherein said at least one anti-VEGFR-2 antibody comprises DC101, and said composition further comprises trifluridine / tipiracil (FTD / TPI).

[0109] Exemplary Embodiment 10b. The method of Exemplary Embodiment 9, wherein said at least one anti-VEGFR-2 antibody is a polyclonal antibody.

[0110] Exemplary Embodiment 11 The method of Exemplary Embodiment 9 or 10, wherein the at least one composition is administered orally to the subject.

[0111] Exemplary embodiment 12. The method of any one of exemplary embodiments 9 to 11, wherein the alternating electric field is applied at a frequency in the range of about 50 kHz to about 10 MHz or in the range of about 50 kHz to about 1 MHz, the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells, and the period during which the alternating electric field is applied is at least about 24 hours.

[0112] Exemplary Embodiment 13. The method of any one of Exemplary Embodiments 9-12, wherein steps (1) and (2) are performed substantially simultaneously.

[0113] Exemplary Embodiment 14. The method of any one of Exemplary Embodiments 9-12, wherein steps (1) and (2) are performed sequentially, in whole or in part, and wherein the at least one composition is administered before the application of the alternating electric field begins.

[0114] Exemplary Embodiment 15. The method of any one of Exemplary Embodiments 9-12, wherein steps (1) and (2) are performed sequentially, in whole or in part, and the at least one composition is administered after the application of the alternating electric field begins.

[0115] Exemplary Embodiment 15a. The method of Exemplary Embodiment 15, wherein said at least one composition is administered after said period of time has elapsed.

[0116] Exemplary Embodiment 15b. The method of Exemplary Embodiment 6, wherein the at least one composition is administered before the period of time during which the alternating electric field is applied has elapsed.

[0117] Exemplary Embodiment 16. The method of any one of Exemplary Embodiments 9-15, wherein steps (1) and (2) are repeated one or more times.

[0118] Exemplary Embodiment 17. The method of any one of Exemplary Embodiments 9-16, 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.

[0119] Exemplary embodiment 18. A method for reducing tumor volume and / or preventing tumor volume growth, wherein the tumor is present in a living body and comprises a plurality of cancer cells, the method comprising: (1) administering to the subject at least one composition, the at least one composition comprising at least one anti-VEGFR-2 antibody; and (2) applying an alternating current electric field to a target area of ​​the subject.

[0120] Exemplary Embodiment 19 The method of Exemplary Embodiment 9, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

[0121] Exemplary Embodiment 19a. The method of Exemplary Embodiment 19, wherein the at least one anti-VEGFR-2 antibody comprises DC101, and the composition further comprises trifluridine / tipiracil (FTD / TPI).

[0122] Exemplary Embodiment 19b. The method of Exemplary Embodiment 9, wherein said at least one anti-VEGFR-2 antibody is a polyclonal antibody.

[0123] Exemplary embodiment 20: The method of exemplary embodiment 18 or 19, wherein the alternating electric field is applied at a frequency in the range of about 50 kHz to about 10 MHz or in the range of about 50 kHz to about 1 MHz, the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells, and the period of time during which the alternating electric field is applied is at least about 24 hours.

[0124] Exemplary embodiment 21. A kit comprising at least one pair of transducer arrays for generating an alternating electric field therebetween upon application of said transducer arrays to at least one cell and / or placement of said transducer arrays on a subject, and at least one composition comprising at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (anti-VEGFR-2 antibody).

[0125] Exemplary Embodiment 22 The kit of Exemplary Embodiment 21, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

[0126] Exemplary Embodiment 22a. The method of Exemplary Embodiment 22, wherein said at least one anti-VEGFR-2 antibody comprises DC101, and said at least one composition further comprises trifluridine / tipiracil (FTD / TPI).

[0127] Exemplary Embodiment 22b. The kit of exemplary embodiment 21 or 22, wherein said at least one anti-VEGFR-2 antibody is a polyclonal antibody.

[0128] Exemplary Embodiment 23 The kit of Exemplary Embodiment 21 or 22, wherein the at least one composition is formulated for oral administration to the subject.

[0129] Exemplary Embodiment 24. A kit described in any one of exemplary embodiments 21 to 23, wherein the at least one pair of transducer arrays is capable of generating an alternating electric field having a frequency in the range of about 50 kHz to about 1 MHz, and / or the at least one pair of transducer arrays is capable of generating an alternating electric field having a field strength of at least about 1 V / cm.

[0130] Exemplary embodiment 25. Use of the kit of any one of claims 21 to 24 in a method for treating cancer, the method comprising the steps of (1) administering at least one composition to the subject, and (2) applying an alternating electric field to a target area of ​​the subject.

[0131] Exemplary embodiment 26. Use of at least one composition comprising at least one anti-VEGFR-2 antibody in a method for treating cancer, the method comprising the steps of: (1) administering the at least one composition to the subject; and (2) applying an alternating current electric field to a target area of ​​the subject.

[0132] Exemplary Embodiment 27 The use of exemplary embodiment 25 or 26, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

[0133] Exemplary Embodiment 28 The use of any one of Exemplary Embodiments 25-27, wherein the at least one composition is administered orally to the subject.

[0134] Exemplary Embodiment 29: The use described in any one of exemplary embodiments 25 to 28, wherein the alternating electric field is applied at a frequency ranging from about 50 kHz to about 1 MHz, the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells, and the duration of application of the alternating electric field is at least about 24 hours.

[0135] Exemplary embodiment 30. The use of any one of exemplary embodiments 25 to 29, wherein the method is further defined as a method of reducing the volume of the tumor and / or preventing an increase in the volume of the tumor, the tumor being present in a living organism and comprising a plurality of cancer cells.

[0136] Exemplary Embodiment 31. A method for reducing the viability of cancer cells, the method comprising: (1) administering to the cancer cells at least one composition, the at least one composition comprising at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (anti-VEGFR-2 antibody); and (2) applying an alternating electric field to the cancer cells.

[0137] Exemplary Embodiment 32 The use or method of any one of embodiments 25 to 31, wherein said method is performed in vivo or ex vivo.

[0138] Exemplary Embodiment 33. The use or method of any one of exemplary embodiments 25 to 32, wherein the alternating electric field is applied at a frequency ranging from about 50 kHz to about 1 MHz, the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells, and the duration of application of the alternating electric field is at least about 24 hours.

[0139] Exemplary Embodiment 34 The use or method of any one of Exemplary Embodiments 25-33, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

[0140] Exemplary Embodiment 34a. The use or method of exemplary embodiment 34, wherein said at least one anti-VEGFR-2 antibody comprises DC101, and said composition further comprises trifluridine / tipiracil (FTD / TPI).

[0141] Exemplary Embodiment 34b. The use or method of exemplary embodiment 31 or 32, wherein said at least one anti-VEGFR-2 antibody is a polyclonal antibody.

[0142] Exemplary Embodiment 35. The use or method of any one of Exemplary Embodiments 25-34, 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.

[0143] Exemplary Embodiment 36. The use or method of any one of Exemplary Embodiments 25-35, wherein steps (1) and (2) are performed substantially simultaneously.

[0144] Exemplary Embodiment 37. The use or method of any one of Exemplary Embodiments 25-36, wherein steps (1) and (2) are performed sequentially, in whole or in part, and wherein the at least one composition is administered before the application of the alternating electric field begins.

[0145] Exemplary Embodiment 38. The use or method of any one of Exemplary Embodiments 25-37, wherein steps (1) and (2) are performed sequentially, in whole or in part, and the at least one composition is administered after the application of the alternating electric field has begun.

[0146] Exemplary Embodiment 39 The use or method of Exemplary Embodiment 38, wherein the at least one composition is administered before the period of time during which the alternating electric field is applied has elapsed.

[0147] Exemplary Embodiment 40 The use or method of Exemplary Embodiment 38, wherein the at least one composition is administered after the period of time has elapsed.

[0148] Exemplary Embodiment 41. The use or method of any one of Exemplary Embodiments 25-40, wherein steps (1) and (2) are repeated one or more times.

[0149] Exemplary Embodiment 42 The use or method of any one of Exemplary Embodiments 25-41, wherein the at least one composition is administered orally to the subject.

[0150] While the present invention describes the concept(s) of the present invention in conjunction with the specific experiments, results, and language set forth below, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, the present invention is intended to embrace all such alternatives, modifications, and variations that fall within the spirit and broad scope of this disclosure.

Claims

1. A kit comprising: at least one pair of transducer arrays for applying 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 comprising at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (anti-VEGFR-2 antibody).

2. 2. The kit of claim 1, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

3. 3. The kit of claim 1 or 2, wherein the at least one composition is formulated for oral administration to the subject.

4. the at least one pair of transducer arrays being capable of generating an alternating electric field having a frequency in the range of about 50 kHz to about 1 MHz; The kit according to any one of claims 1 to 3, wherein the at least one pair of transducer arrays is capable of generating an alternating electric field having a field strength of at least about 1 V / cm.

5. Use of the kit according to any one of claims 1 to 4 in a method for treating cancer, said method comprising: (1) administering said at least one composition to said subject; (2) applying an alternating electric field to a target area of ​​the subject.

6. 1. Use of at least one composition comprising at least one anti-VEGFR-2 antibody in a method for treating cancer, comprising: (1) administering said at least one composition to said subject; (2) applying an alternating electric field to a target area of ​​the subject.

7. The use of claim 6, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

8. 8. The use according to claim 6 or 7, wherein the at least one composition is administered orally to the subject.

9. the alternating electric field is applied at a frequency ranging from about 50 kHz to about 1 MHz; the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells; and The use according to any one of claims 6 to 8, wherein the period during which the AC electric field is applied satisfies at least one of the following conditions: at least about 24 hours.

10. 10. The use of any one of claims 6 to 9, wherein the method is further defined as a method of reducing the volume of the tumor and / or preventing an increase in the volume of the tumor, the tumor being present in a living organism and comprising a plurality of cancer cells.

11. 1. A method of reducing the viability of cancer cells, the method comprising: (1) administering at least one composition to the cancer cells, wherein the at least one composition comprises at least one antibody that specifically binds to vascular endothelial growth factor receptor-2 (anti-VEGFR-2 antibody); (2) applying an alternating electric field to the cancer cells.

12. 12. The method of claim 11, wherein the method is performed in vivo or ex vivo.

13. the alternating electric field is applied at a frequency ranging from about 50 kHz to about 1 MHz; the alternating electric field has an electric field strength of at least about 1 V / cm in at least a portion of the cancer cells; and The use according to any one of claims 6 to 8, wherein the period during which the AC electric field is applied satisfies at least one of the following conditions: at least about 24 hours.

14. The method of any one of claims 11 to 13, wherein the at least one anti-VEGFR-2 antibody is a monoclonal antibody selected from the group consisting of ramucirumab, DC101, and combinations thereof.

15. 15. The method of any one of claims 11 to 14, 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.

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