Compositions and methods for treatment by combination of an alternating electric field and an FGF inhibitor

Abstract

A method of treating a subject in need thereof, comprising the steps of applying an alternating electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time, and administering one or more FGF inhibitors or FGFR inhibitors to the subject in need thereof. A method of enhancing the sensitivity of cells to an alternating electric field, comprising the steps of applying an alternating electric field at a constant frequency to the cells for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby enhancing the sensitivity of the cells to the alternating electric field. A method of enhancing cytotoxicity in cells, comprising the steps of applying an alternating electric field at a constant frequency to the cells for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby enhancing cytotoxicity in the cells. A method of maintaining or enhancing the sensitivity of cells to an alternating electric field, comprising the steps of applying an alternating electric field at a constant frequency to the cells for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining or enhancing the sensitivity of the cells to the alternating electric field. In some embodiments, maintaining or enhancing the sensitivity of cells to an alternating electric field is the same as reducing the resistance of the cells to the alternating electric field.

Description

Technical Field

[0001] Cross - reference to related applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 353,559, filed on June 18, 2022, U.S. Provisional Patent Application No. 63 / 499,842, filed on May 3, 2023, and U.S. Provisional Patent Application No. 63 / 504,093, filed on May 24, 2023, which are hereby incorporated by reference in their entireties.

Background Art

[0002] The fibroblast growth factor (FGF) signaling pathway controls numerous cellular processes such as cell proliferation, apoptosis, angiogenesis, migration, invasion, and metastasis. FGF / FGF receptor (FGFR) exerts physiological functions by regulating downstream targets. Chemical inhibitors, antibodies, and natural products of FGF / FGFR can be used to block the FGF signaling pathway. Therefore, targeting FGF / FGFR may be an effective approach for treating cancer patients in combination with the tumor treatment field (TT field).

Summary of the Invention

[0003] Disclosed is a method of treating a subject in need thereof, the method comprising the steps of applying an alternating electric field at a certain frequency to a target site of a subject in need thereof for a certain period of time, and administering one or more FGF inhibitors or FGFR inhibitors to a subject in need thereof.

[0004] A method of enhancing the sensitivity of cells to an alternating electric field, the method comprising the steps of applying an alternating electric field to the cells at a certain frequency for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby enhancing the sensitivity of the cells to the alternating electric field.

[0005] A method for enhancing intracellular cytotoxicity, the method comprising applying an alternating current electric field to cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby enhancing intracellular cytotoxicity.

[0006] A method for maintaining or enhancing the sensitivity of cells to an alternating current electric field, the method comprising applying an alternating current electric field to cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining or enhancing the sensitivity of the cells to the alternating current electric field. In some embodiments, maintaining or enhancing the sensitivity of the cells to the alternating current electric field is the same as reducing the resistance of the cells to the alternating current electric field.

[0007] Additional advantages of the disclosed methods and compositions are in part illustrated in the following description, in part understood from the description, or may be learned by practice of the disclosed methods and compositions. The advantages of the present invention are realized and achieved by the elements and combinations particularly pointed out in the appended claims. It is to be understood that the foregoing general description and the following detailed description are for purposes of illustration and explanation only and are not intended to limit the claimed invention.

Brief Description of the Drawings

[0008] The accompanying drawings, which are incorporated herein and constitute a part hereof, illustrate some embodiments of the disclosed methods and compositions and, together with the description, serve to explain the principles of the disclosed methods and compositions.

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Modes for Carrying Out the Invention

[0009] The disclosed methods and compositions may be more readily understood by reference to the following detailed description of specific embodiments, the examples included therein, and the figures and the descriptions before and after them.

[0010] It should be understood that the disclosed methods and compositions are not limited to a particular synthesis method, a particular analytical technique, or a particular reagent, unless otherwise specified, and may vary. It should also be understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0011] Materials, compositions, and components that can be used in, combined with, used in the preparation of, or are products of the disclosed methods and compositions are disclosed. These and other materials are disclosed herein, and when combinations, subsets, interactions, groups, etc. of these materials are disclosed, it may not be explicitly disclosed to specifically refer to each of the various individual and collective combinations and permutations of these compounds, but it is understood that each is specifically contemplated and described herein. Thus, if classes of molecules A, B, and C are disclosed, and further an example of a combined molecule A-D with classes of molecules D, E, and F is disclosed, each is considered individually and collectively even if not individually described. Thus, in this example, each of the combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F is specifically contemplated and should be considered disclosed from the disclosure of A, B, and C, D, E, and F, and the example combination A-D. Similarly, these subsets or combinations are specifically contemplated and disclosed. Thus, for example, the subgroups A-E, B-F, and C-E are specifically envisioned and should be considered disclosed from the disclosure of A, B, and C, D, E, and F, and the example combination A-D. This concept applies to all aspects of this application, including but not limited to steps of methods of making and using the disclosed compositions. Thus, if there are various additional steps that can be performed, it is understood that each of these additional steps can be performed in any particular embodiment or combination of embodiments of the disclosed method, and that each such combination is specifically contemplated and should be considered disclosed. A. Definitions

[0012] The disclosed methods and compositions are not limited to the specific methods, protocols, and reagents described, and it is understood that these may vary. It is also understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention, which is limited only by the appended claims.

[0013] Also, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, references to "FGF inhibitor" or "FGFR inhibitor" include plural such inhibitors, and references to "inhibitor" include references to one or more inhibitors and their equivalents known to those of skill in the art.

[0014] As used herein, "target site" refers to a specific site or location present within or on the surface of a subject or patient. For example, "target site" can refer to a cell (e.g., a cancer cell), cell population, organ, tissue, or tumor, but is not limited thereto. Thus, the phrase "target cell" can be used to refer to a target site where the target site is a cell. In some embodiments, the "target cell" can be a cancer cell. In some embodiments, organs that can be target sites include, but are not limited to, the brain. In some embodiments, cells or cell populations that can be target sites or target cells include, but are not limited to, cancer cells (e.g., ovarian cancer cells). In some embodiments, the "target site" can be a target site of a tumor.

[0015] As used herein, the "tumor target site" is a site or location within or on a subject or patient that contains one or more cancer cells, is adjacent to cancer cells, previously contained one or more tumor cells, or is suspected of containing one or more tumor cells. For example, the tumor target site can refer to a site or location within or on the body of a subject or patient who is prone to metastasis. In addition, the target site or tumor target site can refer to the resection site or location of a primary tumor present within or on the surface of the subject or patient. In addition, the target site or tumor target site can refer to a site or location adjacent to the resection of a primary tumor present within or on the body or on the body of the subject or patient.

[0016] As used herein, "alternating electric field" refers to a very low-intensity, directional, medium-frequency alternating electric field that is delivered to a subject, a sample taken from the subject, or a specific location (e.g., a target site such as a cell) within the subject or patient. In some embodiments, the alternating electric field can be in a single direction or multiple directions. In some embodiments, the alternating electric field can be delivered through two pairs of transducer arrays that generate a field perpendicular within the target site. For example, in the case of the Optune® system (an alternating electric field delivery system), one pair of electrodes is placed on the left-right (LR) of the target site and the other pair of electrodes is placed on the anterior-posterior (AP) of the target site. By circulating the electric field between these two directions (LR and AP), it is ensured that the maximum range of cell orientations is targeted.

[0017] As used herein, the "alternating electric field" applied to the tumor target site can be referred to as the "tumor treatment electric field" or "TT field". The TT field has been established as an anti-mitotic cancer therapy because it interferes with the proper assembly of microtubules during metaphase and ultimately destroys cells during anaphase, cytokinesis, or subsequent interphase. The TT field targets solid tumors and is described in U.S. Patent No. 7,565,205, which is hereby incorporated by reference in its entirety as the teaching of the TT field.

[0018] In vivo and in vitro studies have shown that the efficacy of TT field therapy increases as the intensity of the electric field increases. Therefore, optimizing the array placement on a subject to increase the intensity at the subject site or subject cells is a standard approach for the Optune system. Optimization of the array placement may be performed by "rules of thumb" (e.g., placing the array as close as possible to the target site or target cells of the subject), the shape of the patient's body, the dimensions of the target site, and / or measurements representing the position of the target site or cells. The measurements used as input may be obtained from image data. The image data may include any type of visual data, such as, for example, single photon emission computed tomography (SPECT) image data, X-ray computed tomography (X-ray CT) data, magnetic resonance imaging (MRI) data, positron emission tomography (PET) data, data that can be captured by optical devices (e.g., photographic cameras, charge-coupled device (CCD) cameras, infrared cameras, etc.). In certain embodiments, the image data may include 3D data (e.g., point cloud data) obtained from or generated by a 3D scanner. Optimization may depend on understanding how the electric field is distributed within the target site or target cells as a function of the position of the array, and in some aspects, takes into account variations in the distribution of electrical properties within the heads of various patients.

[0019] The term "subject" refers to a subject of administration, such as an animal. Thus, the subject of the disclosed method can be a vertebrate such as a mammal. For example, the subject can be a human. This term does not indicate a particular age or gender. The subject can be used interchangeably with "individual" or "patient". For example, the subject to be administered can mean one that receives an alternating electric field. For example, the subject to be administered can be a patient with ovarian cancer or lung cancer.

[0020] "To treat" means to administer or apply a therapeutic agent, such as an alternating electric field or a vector, to a subject, such as a human or other mammal (e.g., an animal model) suffering from cancer or having an increased susceptibility to developing cancer, in order to prevent or delay the worsening of the effects of a disease or infection, or to partially or completely reverse the effects of cancer. For example, treating a subject suffering from glioblastoma may involve delivering a therapeutic agent to cells within the subject.

[0021] "To prevent" means to minimize or reduce the likelihood that a subject will develop cancer.

[0022] As used herein, the terms "administering" and "administration" refer to any method of directly or indirectly providing an FGF or FGFR inhibitor to a target site in a subject. Such methods are well known to those of skill in the art and include, but are not limited to, oral administration, transdermal administration, inhalation administration, nasal administration, topical administration, intravaginal administration, intraocular administration, intraauricular administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injections such as intravenous administration, intraarterial administration, intramuscular administration, and subcutaneous administration. Administration can be carried out continuously or intermittently. In various embodiments, the formulation can be administered therapeutically, i.e., for treating cancer. Further, in various embodiments, the formulation can be administered prophylactically, i.e., for preventing cancer. In one embodiment, a skilled artisan can determine an effective dosage, an effective schedule, or an effective route of administration for treating a subject. In some embodiments, administration includes exposure or application. Thus, in some embodiments, exposing or applying a target site or a subject to an alternating electric field means subjecting the target site or the subject to an alternating electric field.

[0023] "Any" or "optionally" means that it is not clear whether the event, situation, or material described thereafter occurs or exists, and the description includes both the case where the event, situation, or material occurs or exists and the case where it does not occur or exist.

[0024] In this specification, a range may be expressed as "about" a particular value and / or "about" another particular value. When such a range is expressed, unless otherwise specified in the context, the range from a particular value and / or to another particular value is also specifically contemplated and considered to be disclosed. Similarly, when a value is expressed as an approximation by use of the antecedent "about", unless otherwise specified in the context, that particular value should be understood to form another specifically contemplated embodiment that is disclosed. Further, unless otherwise specified in the context, it is understood that each endpoint of a range is important both in relation to other endpoints and independently of other endpoints. Finally, it is necessary to understand that all individual values and sub-ranges of values included within the explicitly disclosed range are also specifically contemplated and should be considered to be disclosed unless otherwise indicated by the context. The above applies regardless of whether some or all of these embodiments are explicitly disclosed in a particular case.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed methods and compositions belong. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the methods and compositions of the present invention, the particularly useful methods, devices, and materials are as described herein. Publications and the materials they cite that are hereby incorporated by reference are expressly incorporated herein by reference. Nothing in this specification should be construed as an admission that the present invention has no right to antedate such disclosure by virtue of prior invention. Nor should any reference be construed as an admission that such reference constitutes prior art. In the description of the references, the claims of their authors are stated, and the applicant reserves the right to challenge the accuracy and appropriateness of the cited documents. Although numerous publications are referenced herein, it is clear that no reference is admitted that any of these documents constitutes a part of the common general knowledge in the art.

[0026] Throughout the description and claims of this specification, the words "comprise", "comprising", and "comprises", and variations thereof, mean "including but not limited to", and are not intended to (and do not) exclude other additives, components, integers, or steps. In particular, in a method described as including one or more steps or operations, it is specifically contemplated that each step includes (unless a limiting term such as "consisting of" is included in that step). That is, each step is not intended to exclude other additives, components, integers, or steps not recited in that step. B. Alternating electric field

[0027] The methods disclosed herein include an alternating electric field. In some embodiments, the alternating electric field used in the methods disclosed herein is a tumor treatment electric field. In some embodiments, the alternating electric field can vary depending on the cell type or state to which the alternating electric field is applied. In some embodiments, the alternating electric field can be applied via one or more electrodes disposed on a subject's body. In some embodiments, there can be two or more electrode pairs. For example, the array can be disposed on the front / back and sides of the patient and can be used with the systems and methods disclosed herein. In some embodiments, when two pairs of electrodes are used, the alternating electric field can be generated alternately between the electrode pairs. For example, the first electrode pair can be disposed on the front and back of the subject, the second electrode pair can be disposed on either side of the subject, and then the alternating electric field can be applied alternately between the front and back electrodes and then between the left and right electrodes.

[0028] In some embodiments, the frequency of the alternating electric field is from 100 kHz to 500 kHz. In some embodiments, the frequency of the alternating electric field is from 50 kHz to 1 MHz. The frequency of the alternating electric field can be, but is not limited to, 50 - 500 kHz, 100 - 500 kHz, 25 kHz - 1 MHz, 50 - 190 kHz, 25 - 190 kHz, 180 - 220 kHz, or 210 - 400 kHz. In some embodiments, the frequency of the alternating electric field can be an electric field at a frequency of 50 kHz, 100 kHz, 150 kHz, 200 kHz, 250 kHz, 300 kHz, 350 kHz, 400 kHz, 450 kHz, 500 kHz, or any frequency therebetween. In some embodiments, the frequency of the alternating electric field is from about 200 kHz to about 400 kHz, from about 250 kHz to about 350 kHz, and can be about 300 kHz.

[0029] In some embodiments, the electric field strength of the alternating electric field can range from 0.5 to 4 V / cm RMS. In some embodiments, the electric field strength of the alternating electric field can range from 1 to 4 V / cm RMS. In some embodiments, different electric field strengths (e.g., 0.1 to 10 V / cm) can be used. In some embodiments, the electric field strength can be 1.75 V / cm RMS. In some embodiments, the electric field strength is at least 1 V / cm RMS. In some embodiments, the electric field strength can be 0.9 V / cm RMS. In other embodiments, combinations of electric field strengths are applied, for example, by combining two or more frequencies simultaneously or applying two or more frequencies at different times.

[0030] In some embodiments, the alternating electric field can be applied at various intervals ranging from 0.5 hours to 72 hours. In some embodiments, different periods can be used (e.g., 0.5 hours to 14 days). In some embodiments, the application of the alternating electric field can be repeated periodically. For example, the alternating electric field can be applied for 2 hours per day.

[0031] In some embodiments, the exposure may continue over continuous exposure for at least 6 hours, at least 12 hours, at least 24 hours, at least 36 hours, at least 48 hours, or at least 72 hours or more. In some embodiments, the exposure may be the cumulative exposure time of at least 6 hours, at least 12 hours, at least 24 hours, at least 36 hours, at least 48 hours, or at least 72 hours or more. For example, in some embodiments, the exposure can occur at different times over a period of days, weeks, or months. In some embodiments, the patient has the alternating electric field applied for at least 50%, 60%, 70%, 80%, or 90% of the treatment time.

[0032] In some embodiments, the application of the alternating electric field can be continuous or discontinuous. For example, in some embodiments, the application of the alternating electric field can be interrupted for a short time so that the subject can take a shower or take a break from the treatment. In some embodiments, regardless of the short interruption of the application of the alternating electric field, the subject is exposed to the alternating electric field for at least 50%, 60%, 70%, 80%, 90% of the treatment time.

[0033] The disclosed method includes the step of applying one or more alternating electric fields to a cell or a subject. In some embodiments, the alternating electric field is applied to a target site or a tumor target site. When applying an alternating electric field to a cell, this can refer to applying the alternating electric field to the subject constituting the cell. Thus, when applying an alternating electric field to a target site of a subject, the alternating electric field will be applied to the cell. C.FGF / FGFR inhibitor

[0034] As used herein, an FGF inhibitor is a composition that binds to an FGF protein, peptide, or nucleic acid encoding FGF and prevents FGF from interacting or binding with one or more FGFRs. In some embodiments, the FGF inhibitor specifically inhibits or reduces FGF expression. In some embodiments, the FGF inhibitor specifically binds to a nucleic acid encoding FGF. In some embodiments, "specific" means that the inhibitor is selective for FGF or FGFR, as opposed to a non-selective inhibitor that may target multiple receptors. For example, in some embodiments, the disclosed FGF or FGFR inhibitor is selective for FGF or FGFR and does not inhibit a second receptor such as VEGFR. In some embodiments, an inhibitor that specifically inhibits FGF or FGFR may have an IC50 of less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than 1 nM for at least one of FGF1, 2, 3, or 4 or FGFR1, 2, 3, or 4. For example, pemigatinib has a significantly lower IC50 than lenvatinib (a non-selective inhibitor) (Kommalapati et al. Cancers. 2021 June 13; 13(12):2968).

[0035] In some embodiments, the FGF is FGF-21, FGF-19, FGF-7, or FGF-basic. In some embodiments, the FGF inhibitor can be a small molecule, peptide, protein, antibody, or nucleic acid (e.g., siRNA). Examples of FGF inhibitors include, but are not limited to, soluble FGFRs such as soluble FGFR3 and soluble decoy receptors such as FGF-Trap, a soluble decoy receptor fusion protein that binds to FGF-2.

[0036] As used herein, an FGFR inhibitor is a composition that binds to an FGFR protein, peptide, or nucleic acid encoding FGFR and prevents FGFR from interacting or binding with one or more FGFs. In some embodiments, the FGFR inhibitor specifically inhibits or reduces FGFR expression. In some embodiments, the FGFR inhibitor specifically binds to a nucleic acid encoding FGFR. In some embodiments, the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4. In some embodiments, the FGFR inhibitor can be a small molecule, peptide, protein, antibody, or nucleic acid (e.g., siRNA). In some embodiments, the FGFR inhibitor can be one or more of the inhibitors listed in Table 1.

[0037]

Table 1

[0038] Compositions and formulations are disclosed that include one or more FGF inhibitors or FGFR inhibitors, or combinations thereof. In some embodiments, the formulation further includes a pharmaceutically acceptable carrier or diluent. For example, pharmaceutical compositions are disclosed that include an FGF inhibitor or FGFR inhibitor and a pharmaceutically acceptable carrier. For example, pharmaceutical compositions are disclosed that include pemigatinib or AZD4547 and a pharmaceutically acceptable carrier. Pharmaceutical compositions are also disclosed that include an FGF inhibitor or FGFR inhibitor and a pharmaceutically acceptable diluent.

[0039] In some embodiments, the FGF inhibitor or FGFR inhibitor can be administered with a pharmaceutically acceptable carrier and / or diluent in any of the disclosed methods.

[0040] For example, the compositions described herein may include a pharmaceutically acceptable carrier. "Pharmaceutically acceptable" means a material or carrier that, as is well known to those skilled in the art, is selected to minimize degradation of the active ingredient and to minimize any harmful side effects to the subject. Examples of carriers include dimyristoyl phosphatidylcholine (DMPC), phosphate buffered saline, or multilamellar liposomes. For example, PG:PC:cholesterol:peptide or PC:peptide can be used as carriers in the present invention. Other suitable pharmaceutically acceptable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy (19th ed.) ed. A.R. Gennaro, Mack Publishing Company, Easton, PA 1995. Usually, an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to render it isotonic. Other examples of pharmaceutically acceptable carriers include, but are not limited to, saline, Ringer's solution, dextrose solution, etc. The pH of the solution can be from about 5 to about 8, or from about 7 to about 7.5. Further carriers include sustained release formulations such as the semipermeable matrix of a solid hydrophobic polymer containing the composition, which matrix is in the form of shaped articles such as films, stents (implanted in blood vessels during angioplasty), liposomes, or microparticles. For example, it will be apparent to those skilled in the art that a particular carrier may be more preferred depending on the route of administration and the concentration of the composition being administered. These are typically standard carriers for drug administration to humans, including solutions such as sterile water, saline, buffers at physiological pH, etc.

[0041] The pharmaceutical composition may also include a carrier, thickening agent, diluent, buffer, preservative, etc., as long as the intended activity of the polypeptide, peptide, nucleic acid, vector of the present invention is not impaired. In addition to the composition of the present invention, the pharmaceutical composition may also include one or more active ingredients such as antibacterial agents, anti-inflammatory agents, anesthetics, etc. In the method described herein, delivery of the disclosed composition to cells can be effected via various mechanisms. The pharmaceutical composition may be administered in various ways depending on whether local or systemic treatment is desired and on the site to be treated. 1. Delivery of the composition

[0042] Formulations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, emulsions. Examples of non-aqueous solvents include vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and organic esters for injection such as ethyl oleate. Aqueous carriers include water, alcohol / aqueous solutions, emulsions or suspensions (including physiological saline and buffer media). Parenteral carriers include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, fixed oils, etc. Formulations for intravenous administration include bases such as water and nutritional supplements, electrolyte supplements (e.g., those based on Ringer's dextrose). For example, preservatives and other additives such as antibacterial agents, antioxidants, chelating agents, inert gases may also be present.

[0043] Formulations for topical administration may include ointments, lotions, creams, gels, eye drops, suppositories, sprays, liquids, powders, etc. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickening agents, etc. may be required or desirable.

[0044] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersion aids, or binders may be desirable. Some of the compositions may be administered as pharmaceutically acceptable acid or base addition salts formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, or by reaction with inorganic bases such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as monoalkylamine, dialkylamine, trialkylamine, arylamine, substituted ethanolamine. E. Treatment methods

[0045] Disclosed is a method for treating a subject in need thereof, comprising the steps of applying an alternating electric field at a certain frequency to a target site of the subject in need thereof for a certain period of time, and administering one or more FGF inhibitors or FGFR inhibitors to the subject in need thereof. Also disclosed is a method for treating a subject in need thereof, comprising the steps of applying an alternating electric field at a certain frequency to a target site of the subject in need thereof for a certain period of time, and administering to the subject in need thereof a composition comprising one or more FGF inhibitors or FGFR inhibitors.

[0046] In some embodiments, the subject in need thereof has cancer. In some embodiments, the subject in need thereof has a mesothelioma, ovarian cancer, or lung cancer. In some embodiments, the subject in need thereof has a brain tumor, pancreatic cancer, breast cancer, or colon cancer.

[0047] In some embodiments, the target site comprises one or more cancer cells. In some embodiments, the target site comprises one or more mesothelioma cells, ovarian cancer cells, lung cancer cells, brain cancer cells, pancreatic cancer cells, breast cancer cells, or colon cancer cells.

[0048] In some embodiments, the alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF inhibitors or FGFR inhibitors. In some embodiments, the step of applying the alternating electric field is initiated at least 1 hour before the FGF inhibitor or FGFR inhibitor. In some embodiments, the step of applying the alternating electric field is initiated at least 30 minutes before the FGF inhibitor or FGFR inhibitor. In some embodiments, applying the alternating electric field simultaneously may mean applying it within 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 minutes before or after the administration of the FGF inhibitor or FGFR inhibitor. In some embodiments, the alternating electric field is applied and the FGF inhibitor or FGFR inhibitor may be administered at intervals of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours from each other. In some embodiments, applying the alternating electric field simultaneously with the administration of one or more FGF inhibitors or FGFR inhibitors includes applying the alternating electric field while one or more FGF inhibitors or FGFR inhibitors are active in a specific state in the subject's body.

[0049] In some embodiments, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (target or non-target), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, during surgery or biopsy), or by inhalation.

[0050] In some embodiments, the frequency of the alternating electric field is between 50 kHz and 1 MHz. In some embodiments, the frequency of the alternating electric field is 150 kHz or 200 kHz. In some embodiments, the alternating electric field can be any within the ranges described herein.

[0051] In some embodiments, the alternating electric field has an electric field strength of 0.1 to 10 V / cm RMS. In some embodiments, the alternating electric field has an electric field strength of 0.5 to 4 V / cm RMS. In some embodiments, the alternating electric field has an electric field strength of 0.9 V / cm RMS. In some embodiments, the alternating electric field has any of the electric field strengths described herein.

[0052] In some embodiments, the disclosed treatment method may further comprise administering a cancer therapeutic agent. In some embodiments, the cancer therapeutic agent is a known cancer therapeutic agent other than an FGF or FGFR inhibitor. For example, the cancer therapeutic agent can be, but is not limited to, chemotherapy, radiation therapy, immunotherapy, or hormone therapy.

[0053] In some embodiments, the alternating electric field is applied before, after, or simultaneously with the administration of the cancer therapeutic agent. In some embodiments, the FGF or FGFR inhibitor is applied before, after, or simultaneously with the administration of the cancer therapeutic agent. In some embodiments, one or more FGF or FGFR inhibitors and the cancer therapeutic agent are administered simultaneously, and the alternating electric field is applied before or after the FGF or FGFR inhibitor and the cancer therapeutic agent.

[0054] In some embodiments, the steps of the disclosed method must be performed in the order in which the steps are listed. For example, a method of treating a subject in need thereof, comprising first applying an alternating electric field at a certain frequency to a target site of the subject in need thereof for a certain period of time, and then administering to the subject in need thereof one or more FGF inhibitors or FGFR inhibitors, or a composition comprising one or more FGF inhibitors or FGFR inhibitors, is disclosed.

[0055] Alternatively, the steps of the disclosed method may be performed in an order different from the recited steps. For example, a method of treating a subject in need thereof, comprising first administering to the subject in need thereof one or more FGF inhibitors or FGFR inhibitors, or a composition comprising one or more FGF inhibitors or FGFR inhibitors, and then applying an alternating electric field at a certain frequency to a target site of the subject in need thereof for a certain period of time, is disclosed. In some embodiments, first administering one or more FGF inhibitors or FGFR inhibitors, or a composition comprising one or more FGF inhibitors or FGFR inhibitors, may increase the sensitivity of cells in the subject to an alternating electric field.

[0056] In some embodiments, the steps of the disclosed method may be performed in any order. F. Methods of affecting cells

[0057] In some embodiments, affecting cancer cells may mean increasing the sensitivity of the cells to an alternating electric field, increasing the cytotoxicity within the cells, or enhancing or maintaining the sensitivity to an alternating electric field.

[0058] A method of increasing the sensitivity of cells to an alternating electric field, comprising applying an alternating electric field to the cells at a certain frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby increasing the sensitivity of the cells to an alternating electric field, is disclosed.

[0059] A method of increasing the cytotoxicity within cells, comprising applying an alternating electric field to the cells at a certain frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby increasing the cytotoxicity within the cells, is disclosed.

[0060] A method for maintaining or enhancing the sensitivity of cells to an alternating electric field, comprising the steps of applying an alternating electric field to the cells at a constant frequency for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining or enhancing the sensitivity of the cells to the alternating electric field. In some embodiments, maintaining or enhancing the sensitivity of cells to an alternating electric field is the same as reducing the resistance of cells to the alternating electric field. Accordingly, a method for reducing the resistance of cells to an alternating electric field, comprising the steps of applying an alternating electric field to the cells at a constant frequency for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby reducing the resistance of the cells to the alternating electric field, is also disclosed.

[0061] A method for maintaining or enhancing the sensitivity of cells to an alternating electric field, comprising the steps of applying an alternating electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time, and administering one or more FGF inhibitors or FGFR inhibitors to a subject in need thereof, thereby maintaining or enhancing the sensitivity of the cells to the alternating electric field. Accordingly, a method for reducing the resistance of cells within a cell to an alternating electric field, comprising the steps of applying an alternating electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time, and administering one or more FGF inhibitors or FGFR inhibitors to a subject in need thereof, thereby reducing the resistance of the cells within the cell to the alternating electric field, is also disclosed.

[0062] In some embodiments of the method for maintaining or enhancing sensitivity, or reducing resistance, after the step of applying the alternating electric field and before the step of contacting or administering the FGF or FGFR inhibitor, a step of detecting an increase in FGF expression in the subject or cell is performed.

[0063] In some embodiments, the methods disclosed herein may further comprise applying an alternating electric field at a constant frequency to the cells for a period of time after the first round of alternating electric field and the FGF inhibitor or FGFR inhibitor has been applied or administered to the subject or cells. For example, the methods disclosed herein are methods for maintaining or enhancing the sensitivity of cells to an alternating electric field, the method comprising applying an alternating electric field at a constant frequency to the target site of a subject in need thereof for a period of time, and administering one or more FGF inhibitors or FGFR inhibitors to a subject in need thereof, thereby maintaining or enhancing the sensitivity of the cells to the alternating electric field, and further comprising applying an alternating electric field at a constant frequency to the target site of a subject in need thereof for a period of time after the sensitivity of the cells to the alternating electric field has been maintained or enhanced.

[0064] In some embodiments, the contacting or administering step is performed at a time when the response of the cells or subject to the alternating electric field has decreased. In some embodiments, the contacting or administering step is performed 3, 4, 5, 6, 7, 8, 9, or 10 days after applying the alternating electric field. In some embodiments, the detection of an increase in FGF expression is determined before contacting the cells with FGF or an FGF inhibitor, or before administering FGF or an FGF inhibitor to the subject. In some embodiments, the detection of FGF expression is performed before applying the alternating electric field to obtain a baseline reading, and then detected again at least 1, 2, 3, 4, 5, 6, or 7 days, or at least 1, 2, 3, or 4 weeks after applying the alternating electric field. In some embodiments, the determination of whether the response of the subject to the alternating electric field has decreased can be measured by analyzing FGF downstream signaling. For example, whether the response of the subject to the alternating electric field has decreased can be measured by determining the presence of FGF in the serum, determining the presence of pAkt in a biopsy, or determining the absence of a response to tumor shrinkage (or tumor growth).

[0065] In some embodiments, the disclosed method can be performed directly on a subject. For example, the method can be performed directly on a subject in need thereof, and the subject is a living body. In some embodiments, the disclosed method can be performed on a sample obtained from a subject. In some embodiments, the disclosed method can be performed outside the subject, and thus, the cells can be in vitro. In some embodiments, the disclosed method can be performed inside the subject, and thus, the cells can be present within the subject. In some embodiments, the cells (in vitro or in vivo) can be mesothelioma cells, ovarian cancer cells, lung cancer cells, brain cancer cells, pancreatic cancer cells, breast cancer cells, cancer cells, or colon cancer cells. In some embodiments, the cells can be from cell lines including, but not limited to, cell line A2780 or H1299.

[0066] In some aspects, the steps of the disclosed method must be performed in the order in which the steps are listed. For example, a method of treating a subject in need thereof, comprising first applying an alternating electric field at a certain frequency to a target site of the subject in need thereof for a certain period of time, and then administering to the subject in need thereof one or more FGF inhibitors or FGFR inhibitors, or a composition comprising one or more FGF inhibitors or FGFR inhibitors, is disclosed.

[0067] Alternatively, the steps of the disclosed method can be performed in an order different from the recited steps. For example, a method of treating a subject in need thereof, comprising first administering to the subject in need thereof one or more FGF inhibitors or FGFR inhibitors, or a composition comprising one or more FGF inhibitors or FGFR inhibitors, and then applying an alternating electric field at a certain frequency to a target site of the subject in need thereof for a certain period of time, is disclosed. In some embodiments, administering first one or more FGF inhibitors or FGFR inhibitors, or a composition comprising one or more FGF inhibitors or FGFR inhibitors, can enhance the sensitivity of cells in the subject to the alternating electric field.

[0068] In some embodiments, the steps of the disclosed method can be performed in any order. G. Kit

[0069] The above materials and other materials can be packaged together in any suitable combination as a kit useful for performing or assisting in the performance of the disclosed method. A kit is useful if the kit components within a particular kit are designed and adapted to be used together in the disclosed method. For example, a kit is disclosed that includes one or more FGF or FGFR inhibitors and one or more materials for delivering an alternating electric field, such as the Optune system. The material for delivering the alternating electric field includes a signal generator and a pair of electrodes. In some embodiments, the material for delivering the alternating electric field can include a signal generator (e.g., an alternating signal generator), one or more pairs of electrode arrays where each electrode array includes two or more electrodes, or a controller. In some embodiments, the material for delivering the alternating electric field can include the Optune system or one or more of its components.

[0070] In some embodiments, the material for delivering an alternating electric field can include a system that includes one or more of the elements of the system shown in FIG. 1. FIG. 1 is a block diagram of an exemplary system for transmitting an alternating electric field. The system includes an alternating signal generator 20 designed to generate first and second alternating outputs, for example, at a frequency of 50 to 500 kHz. When using the system to apply an alternating electric field to the human body, the first alternating output is applied across a first electrode pair 10L and 10R disposed on the left and right of the cancer cells, and the second alternating output is applied across a second electrode pair 10A and 10P disposed in front of and behind the cancer cells. The alternating signal generator 20 can also be used to apply an alternating electric field to in vitro cultures by applying the first alternating output to electrodes disposed on the left and right walls of the dish (or a dedicated Inovitro™ dish), and the second alternating output to electrodes disposed on the front and back walls of the dish (or a dedicated Inovitro™ dish). In either case, the voltage generated by the alternating signal generator 20 must be sufficient to induce an electric field of at least 1 V / cm in at least some of the cancer cells. In some embodiments, the voltage generated by the alternating signal generator 20 must be sufficient to induce an electric field of 1 V / cm to 10 V / cm in at least some of the cancer cells.

[0071] In some embodiments, (a) the first alternating output is applied to the L / R electrodes at sub-time intervals of 1 second, (b) the second alternating output is applied to the A / P electrodes at sub-time intervals of 1 second, and the two-step sequence (a) and (b) is repeated during the treatment period. The alternating signal generator 20 can be configured to generate the first and second alternating outputs such that the amplitude and / or frequency of the first and second alternating outputs change in response to the state of at least one control input.

[0072] Controller 30 continuously transmits control signals to at least one control input at sub - intervals of 1 second. In FIG. 1, the controller 30 and the alternating - current signal generator 20 are shown as two different blocks, but these two blocks may be integrated into a single hardware device. The details of the structure of the controller 30 and the nature of the control signals vary depending on the design of the alternating - current signal generator 20. In one example, the design of the alternating - current signal generator 20 is similar to the alternating - current signal generator described in U.S. Patent No. 9,910,453, the entire text of which is incorporated herein by reference. This particular alternating - current signal generator includes two output channels (i.e., the first channel for L / R and the second channel for A / P).

[0073] In some embodiments, kits are disclosed that include one or more FGF and FGFR inhibitors, materials for delivering an alternating electric field, and additional therapeutic agents. For example, kits are disclosed that include aprepitant, auranofin, captopril, celecoxib, disulfiram, itraconazole, minocycline, ritonavir, sertraline, one or more FGF and FGFR inhibitors, and one or more materials for delivering an alternating electric field such as the Optune system. In some aspects, the kit may also include temozolomide. Examples A. Example 1 1. Overview

[0074] The FGF / FGFR signaling pathway plays a role in the development and progression of various cancers. The FGF signaling pathway controls numerous cellular processes such as cell proliferation, apoptosis, angiogenesis, migration, invasion, and metastasis. FGF / FGFR can be regulated by Notch, N - CAM, miRNA, synthetic compounds, antibodies, and natural substances. FGF / FGFR exerts its physiological functions through the regulation of downstream targets (such as Ras, PI3K / AKT, ERK, NF - kB, VEGF, etc.). Therefore, targeting FGF / FGFR can be an effective approach for the treatment of cancer patients.

[0075] FGF / FGFR exerts its physiological functions by regulating major downstream signaling pathways such as RAS / MAPK and PI3K / AKT / mTOR. However, FGF / FGFR can be blocked by chemical inhibitors, antibodies, receptor decoys, and natural substances of FGF / FGFR. Therefore, targeting FGF / FGFR can be an effective approach for the treatment of various cancers, such as female genital system cancer patients. Secreted FGF binds to one of the four transmembrane receptors (FGFR1, FGFR2, FGFR3, and FGFR4) with an intracellular tyrosine kinase domain in a 2:2:2 HSPG-FGF-FGF receptor ratio. Ligand binding specificity is determined by different expression patterns of FGF, FGF receptors, and glycosaminoglycan structures, different receptor binding abilities, the requirement for specific cofactors such as the Cripto family proteins, and alternative splicing of FGF receptors, resulting in the generation of two different versions (b or c) of the extracellular Ig-like domain III. When the HSPG ligand binds to the FGF receptor, the receptor homodimerizes, the cytoplasmic intracellular kinase domain of the receptor is activated, and adapter proteins such as FRS2, GRB2, Shb, and Shc are recruited and docked. These adapter proteins then activate multiple downstream signaling pathways, including the Ras-MAPK pathway, Jak-STAT pathway, PI3-Kinase-Akt pathway, PLC gamma pathway, p38, and JNK MAPK pathways. Through these signaling pathways, FGF promotes basic cell processes such as survival, proliferation, differentiation, and motility. Therefore, targeting FGF can contribute to the treatment of cancer either alone or in combination with other known cancer treatments such as TT fields.

[0076] Targeting FGF / FGFR has been shown to enhance the antitumor effect of the tumor treatment field (TT field). 2. Materials and Methods i. Cell Culture Model

[0077] The human cell lines A2780 (ovarian cancer) and H1299 (non-small cell lung cancer) were obtained from the American Type Culture Collection (ATCC). The cells were cultured in a humidified incubator at 37°C with 5% CO2 using a medium supplemented with 10% (v / v) fetal bovine serum (FBS) and streptomycin (50 μg / ml). The medium and supplements were purchased from Biological Industries (Beit Haemek). ii. Mouse model

[0078] An orthotopic model of ovarian cancer mice was created. 5000 / 5 μl of MOSE-L-ffl cancer cells were injected into 12-week-old female mice. iii. In vitro a. In vitro system

[0079] Using the inovitro (trademark) system, a TT field (1.7 V / cm RMS) was applied to A2780 cells at 200 kHz and to H1229 cells at 150 kHz. b. Cell count

[0080] Cytotoxicity was determined by counting the cells using a Macsquant (Miltenyi Biotec) flow cytometer and was shown as the percentage of cells relative to the control. iv. In vivo

[0081] Mice were treated with a TT field for 10 days using the INOVIVO system (manufactured by Novocure Ltd., Haifa, Israel). The mice were sacrificed, blood samples were collected into designated serum tubes, centrifuged at 1000 g for 15 minutes, and stored at -20°C. v. Analysis and quantification of cytokine array kit

[0082] Using a cytokine array kit (R&D Systems ARY005B for human-derived samples and ARY028 for mouse-derived samples), 105 cytokines in each human cell line-conditioned medium (CM) / mouse serum sample were simultaneously detected and collected according to the manufacturer's instructions. The array membrane was incubated in blocking buffer at room temperature for 1 hour (all reagents are supplied with the array kit). Before mixing with the reconstituted detection antibody cocktail, the samples were incubated overnight on the membrane at 4°C on a rocking platform shaker. The following steps were all performed at room temperature, and in all washing steps, the membrane was washed three times for 10 minutes with 1× wash buffer. After incubation with the detection antibody, the membrane was washed and incubated with streptavidin-conjugated horseradish peroxidase (1:2000) on a rocking platform shaker for 30 minutes. Unbound reagents were washed away, and the membrane was incubated with chemiluminescent detection reagent for 1 minute. To compare and evaluate the changes after TT field treatment with the control group, the pixel density of the spots was measured using R&D Systems quantification software (Quick Spot), and the results were analyzed as follows: [(protein pixel density of TT field-treated sample)÷(protein pixel density of control sample)]. 3. Results

[0083] Figure 2 shows an example of the cytotoxic effect after TT field treatment at the optimal cytotoxic frequency for 3 days. To characterize the secreted cytokine profile after TT field application, a series of in vitro experiments were performed. Cells were treated at the cytotoxic frequency for 72 hours, and the changes in secreted protein expression were evaluated using a cytokine array assay. Three days after TT field treatment, the cell numbers of both A2780 and H1299 cells decreased significantly.

[0084] After applying the TT field at the optimal cytotoxic frequency to A2780 (200 kHz) and H1299 (150 kHz) cell lines for 3 days, the expression of FGF family growth factors, especially FGF-basic, FGF-7, and FGF-19, was upregulated (see Figures 3A and 3B).

[0085] Figure 4 shows that the expression of FGF-21 is upregulated after applying the TT field to an ovarian cancer mouse model for 10 days.

[0086] Based on these results, by inhibiting / blocking FGF / FGFR signaling, it is possible to reduce the adverse effects of FGF / FGFR signaling while enhancing or maintaining the effectiveness of the TT field. B. Example 2 1. Background

[0087] Glioblastoma multiforme (GBM) is a malignant primary brain tumor with a poor prognosis despite standard treatment combining radiotherapy and temozolomide according to the Stupp protocol. Recently, a new treatment method called tumor treatment field (TT field) has been approved for clinical treatment as a monotherapy for recurrent GBM and as a combination therapy with adjuvant therapy after chemoradiotherapy. Furthermore, the TT field has been shown to act additively or synergistically with radiation, and good tolerance of this combination has been shown in pilot clinical trials, enabling the development of the randomized Trident trial. However, even when combined with the TT field, GBM remains a fatal disease resistant to treatment. One cause of GBM resistance is the presence of cancer stem cells (GSCs). This cell pool is involved in the resistance of tumor cells to treatment and the proliferation of tumor-initiating cells that subsequently promote tumor regrowth. Their maintenance is promoted by hypoxia. Studying the mechanism of GSC resistance to the TT field is a major challenge to address.

[0088] FGFR1 has been demonstrated to control the radioresistance of glioblastoma and glioblastoma stem cells (Gouaze-Andersson et al., Cancer Res. 2016; Gouaze-Andersson et al. Oncotarget 2018), and inhibition by FGFR inhibitors has been shown to lead to radiosensitization of glioblastoma (Ader et al, Eur. J. Cancer 2014). Furthermore, clinical data have shown that GBM patients with high FGFR1 levels have a shorter time to progression and a shorter overall survival after radiotherapy (Ducassou et al., Eur J Cancer 2013). Also, GBM patients expressing a five-gene signature including α6 integrin, ZEB1 / YAP1, FGFR1, and FOXM1 within the tumor have been shown to have a significantly shorter overall survival (Kowalski-Chauvel et al., Cancers 2019 11(3)). These data indicate that targeting FGFR1 can enhance the therapeutic effect of glioblastoma. In this study, the resistance mechanism of GBM stem cells to TT fields, particularly the FGFR1 pathway, was investigated. 2. Study design and methods

[0089] This study focused on the involvement of FGFR1 in the control of sensitivity to TT fields. This study was conducted using several glioblastoma stem cells (GSCs) that were fully characterized in vitro and in vivo, obtained from the clinical trial STEMRI (NCT01872221) at the Institut Claudius Regaud. The inventors express their gratitude to the Institut Claudius Regaud for providing the glioblastoma stem cells used in these experiments.

[0090] Figure 5 shows that TT field treatment induces different levels of cytotoxic effects that can distinguish three GSC groups sensitive, moderately sensitive, and resistant to TT fields. Figure 6 shows that FGFR1 RNA expression is increased by TT fields in GSCs. FGFR1 expression increased in both TTF-sensitive and -resistant GSCs.

[0091] Figure 7 shows the expression level of FGFR1 as a response to the TT field in resistant cells (SRC2) and sensitive cells (SRC1), and its inhibition using pemigatinib, an FGFR1 inhibitor.

[0092] Figure 8 shows that FGFR1 inhibitors can make all glioblastoma cell lines, including cells previously resistant to the TT field, sensitive to the TT field. The mechanism by which radiation induces cell death can be the result of DNA damage leading to apoptosis, and the TT field is also known to cause DNA damage and reduce DNA repair mechanisms. The addition of FGFR inhibitors reduces radiosensitivity. Concurrent treatment with FGFR inhibition increases sensitivity to the TT field, indicating a synergistic effect due to increased secretion of FGF family members in serum and conditioned medium after TT field application, indicating a decrease in cell sensitivity to the TT field.

[0093] These results indicate that it is possible to distinguish between the sensitivity and resistance of GSCs to the TT field and enhance the sensitivity of glioblastoma cells to the TT field by inhibiting FGFR1. Exemplary embodiments

[0094] An example of many embodiments described herein is a method of treating a subject in need thereof, the method comprising applying an alternating electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time, and administering to the subject in need thereof one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors.

[0095] In an example of many embodiments described herein, the subject is alive.

[0096] In an example of many embodiments described herein, the subject has a mesothelioma, ovarian cancer, or lung cancer.

[0097] In one example of many embodiments described herein, the target site comprises one or more cancer cells.

[0098] In one example of many embodiments described herein, an alternating current electric field is applied before, after, or simultaneously with the administration of one or more FGF or FGFR inhibitors.

[0099] In one example of many embodiments described herein, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (target or non-target), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, intraoperative, or during biopsy), or by inhalation.

[0100] One example of many embodiments described herein is a method of enhancing the sensitivity of cells to an alternating current electric field, the method comprising applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating current electric field.

[0101] One example of many embodiments described herein is a method of enhancing the sensitivity of cells to an alternating current electric field, the method comprising applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating current electric field, wherein the cells are within a subject.

[0102] An example of many embodiments described herein is a method of enhancing the sensitivity of cells to an alternating current electric field, the method comprising applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating current electric field, and the method is performed on the cells in vitro.

[0103] An example of many embodiments described herein is a method of enhancing cytotoxicity within cells, the method comprising applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing cytotoxicity within the cells.

[0104] An example of many embodiments described herein is a method of enhancing cytotoxicity within cells, the method comprising applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing cytotoxicity within the cells, and the cells are within a subject.

[0105] An example of many embodiments described herein is a method of enhancing cytotoxicity within cells, the method comprising applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing cytotoxicity within the cells, and the method is performed on the cells in vitro.

[0106] In one example of many embodiments described herein, the FGF inhibitor inhibits or reduces FGF expression.

[0107] In one example of many embodiments described herein, the FGF inhibitor blocks the upregulation of FGF expression in response to an alternating current electric field.

[0108] In one example of many embodiments described herein, the FGF inhibitor prevents FGF from interacting or binding with one or more fibroblast growth factor receptors (FGFRs).

[0109] In one example of many embodiments described herein, the FGFR inhibitor prevents FGFR from interacting or binding with one or more FGFs.

[0110] In one example of many embodiments described herein, the FGFR inhibitor inhibits one or more fibroblast growth factor receptors (FGFRs).

[0111] In one example of many embodiments described herein, the FGF is FGF-21, FGF-19, FGF-7, FGF-basic.

[0112] In one example of many embodiments described herein, the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

[0113] In one example of many embodiments described herein, the FGFR inhibitor can be one or more of the inhibitors in Table 1.

[0114] In one example of many embodiments described herein, the cancer cells are ovarian cancer or lung cancer cells.

[0115] In one example of many embodiments described herein, the frequency of the alternating current electric field is 50 kHz to 1 MHz.

[0116] In one example of many embodiments described herein, the frequency of the alternating current electric field is 150 kHz or 250 kHz.

[0117] In one example of many embodiments described herein, the alternating current electric field has an electric field strength of 0.5 to 4 V / cm RMS.

[0118] In one example of many embodiments described herein, the alternating current electric field has an electric field strength of 0.9 V / cm RMS.

[0119] In one example of many embodiments described herein, the method further includes the step of administering a cancer therapeutic agent.

[0120] One example of many embodiments described herein is a method of maintaining the sensitivity of cells to an alternating current electric field, the method including applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating current electric field.

[0121] One example of many embodiments described herein is a method of maintaining the sensitivity of cells to an alternating current electric field, the method including applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells in a subject to the alternating current electric field.

[0122] One example of many embodiments described herein is a method of maintaining the sensitivity of cells to an alternating current electric field, the method including applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating current electric field, and this method is performed on the cells in vitro.

[0123] One example of many embodiments described herein is a method of maintaining the sensitivity of a subject to an alternating current electric field, the method including applying an alternating current electric field to a target site of the subject at a constant frequency for a certain period of time and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to the subject in need thereof, thereby maintaining the sensitivity of the cells to the alternating current electric field.

[0124] An example of many embodiments described herein is a method of maintaining the sensitivity of cells to an alternating current electric field, the method comprising applying an alternating current electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof, thereby maintaining the sensitivity of the cells to the alternating current electric field.

[0125] An example of many embodiments described herein is a method of maintaining the sensitivity of a subject to an alternating current electric field, the method comprising applying an alternating current electric field at a constant frequency to a target site of the subject for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof, thereby maintaining the sensitivity of the subject or cells (the subject is alive) to the alternating current electric field.

[0126] In an example of many embodiments described herein, the method further comprises detecting an increase in FGF expression in the subject or cells after applying the alternating current electric field and before administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof.

[0127] In an example of many embodiments described herein, the contact is made when the response of the cells to the alternating current electric field decreases.

[0128] In an example of many embodiments described herein, the contact with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor F receptor (FGFR) inhibitors is made 3, 4, 5, 6, 7, 8, 9, or 10 days after the application of the alternating current electric field.

[0129] In an example of many embodiments described herein, the contact is made when the response of the subject to the alternating current electric field decreases.

[0130] In one example of many embodiments described herein, the contact is made 3, 4, 5, 6, 7, 8, 9, or 10 days after step a) is performed.

[0131] In one example of many embodiments described herein, an alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF or FGFR inhibitors.

[0132] In one example of many embodiments described herein, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (targeted or non-targeted), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, intraoperative, or during biopsy), or by inhalation.

[0133] In one example of many embodiments described herein, an FGF inhibitor inhibits or reduces FGF expression.

[0134] In one example of many embodiments described herein, an FGF inhibitor blocks the upregulation of FGF expression in response to an alternating electric field.

[0135] In one example of many embodiments described herein, an FGF inhibitor prevents FGF from interacting or binding to one or more fibroblast growth factor receptors (FGFRs).

[0136] In one example of many embodiments described herein, an FGFR inhibitor prevents FGFR from interacting or binding to one or more FGFs.

[0137] In one example of many embodiments described herein, an FGFR inhibitor blocks one or more fibroblast growth factor receptors (FGFRs).

[0138] In one example of many embodiments described herein, the FGF is FGF-21, FGF-19, FGF-7, or FGF-basic.

[0139] In one example of many embodiments described herein, the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

[0140] In one example of many embodiments described herein, the FGFR inhibitor can be one or more of the inhibitors in Table 1.

[0141] In one example of many embodiments described herein, the frequency of the alternating electric field is 50 kHz to 1 MHz.

[0142] In one example of many embodiments described herein, the frequency of the alternating electric field is 150 kHz or 250 kHz.

[0143] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.5 to 4 V / cm RMS.

[0144] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.9 V / cm RMS.

[0145] In one example of many embodiments described herein, the method further includes the step of administering a cancer therapeutic agent.

[0146] One example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of treating a subject in need thereof, the method comprising applying an alternating electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time and administering a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor to a subject in need thereof, a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0147] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing the sensitivity of cells to an alternating current electric field, the method comprising the steps of applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating current electric field, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0148] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing the sensitivity of cells to an alternating current electric field, the method comprising the steps of applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating current electric field, and the cells are in a subject, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0149] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing the sensitivity of cells to an alternating current electric field, the method comprising the steps of applying an alternating current electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating current electric field, and the method is performed on the cells in vitro, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0150] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing cytotoxicity within a cell, the method comprising applying an alternating electric field to the cell at a certain frequency for a certain period of time and contacting the cell with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the cytotoxicity within the cell. An example of an embodiment is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing cytotoxicity within a cell.

[0151] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing cytotoxicity within a cell, the method comprising applying an alternating electric field to the cell at a certain frequency for a certain period of time and contacting the cell with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the cytotoxicity within the cell, wherein the cell is within a subject. A fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0152] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of enhancing cytotoxicity within a cell, the method comprising applying an alternating electric field to the cell at a certain frequency for a certain period of time and contacting the cell with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the cytotoxicity within the cell, wherein the method is performed on the cell in vitro. A fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0153] In one example of many embodiments described herein, the subject is alive.

[0154] In one example of many embodiments described herein, the subject has a mesothelioma, ovarian cancer, or lung cancer.

[0155] In one example of many embodiments described herein, the target site contains one or more cancer cells.

[0156] In one example of many embodiments described herein, an alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF or FGFR inhibitors.

[0157] In one example of many embodiments described herein, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (target or non-target), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, intraoperative, or during biopsy), or by inhalation.

[0158] In one example of many embodiments described herein, the FGF inhibitor inhibits or reduces FGF expression.

[0159] In one example of many embodiments described herein, the FGF inhibitor blocks the upregulation of FGF expression in response to an alternating electric field.

[0160] In one example of many embodiments described herein, the FGF inhibitor prevents FGF from interacting or binding with one or more fibroblast growth factor receptors (FGFRs).

[0161] In one example of many embodiments described herein, the FGFR inhibitor prevents FGFR from interacting or binding with one or more FGFs.

[0162] In one example of many embodiments described herein, the FGFR inhibitor blocks one or more fibroblast growth factor receptors (FGFRs).

[0163] In one example of many embodiments described herein, the FGF is FGF-21, FGF-19, FGF-7, FGF-basic.

[0164] In one example of many embodiments described herein, the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

[0165] In one example of many embodiments described herein, the FGFR inhibitor can be one or more of the inhibitors in Table 1.

[0166] In one example of many embodiments described herein, the cancer cells are ovarian cancer or lung cancer cells.

[0167] In one example of many embodiments described herein, the frequency of the alternating electric field is 50 kHz to 1 MHz.

[0168] In one example of many embodiments described herein, the frequency of the alternating electric field is 150 kHz or 250 kHz.

[0169] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.5 to 4 V / cm RMS.

[0170] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.9 V / cm RMS.

[0171] In one example of many embodiments described herein, the method further includes the step of administering a cancer therapeutic agent.

[0172] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of maintaining the sensitivity of cells to an alternating electric field, the method comprising applying an alternating electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating electric field, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0173] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of maintaining the sensitivity of cells to an alternating electric field, the method comprising applying an alternating electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating electric field, and the cells are within a subject, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0174] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of maintaining the sensitivity of cells to an alternating electric field, the method comprising applying an alternating electric field to the cells at a constant frequency for a certain period of time and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating electric field, and the method is performed on the cells in vitro, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0175] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of maintaining a subject's sensitivity to an alternating electric field, the method comprising applying an alternating electric field at a constant frequency to a target site of the subject for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof, thereby maintaining the sensitivity of cells to the alternating electric field, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0176] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of maintaining a cell's sensitivity to an alternating electric field, the method comprising applying an alternating electric field at a constant frequency to a target site of a subject in need thereof for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof, thereby maintaining the sensitivity of cells to the alternating electric field, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0177] An example of many embodiments described herein is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor for use in a method of maintaining a subject's sensitivity to an alternating electric field, the method comprising applying an alternating electric field at a constant frequency to a target site of the subject for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof, thereby maintaining the sensitivity of the subject or cells (the subject is alive) to the alternating electric field, which is a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor.

[0178] In one example of many embodiments described herein, the method further includes detecting an increase in FGF expression in a subject or cells after applying an alternating current electric field and before administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to a subject in need thereof.

[0179] In one example of many embodiments described herein, the contact is made at a time when the response of the cells to the alternating current electric field has decreased.

[0180] In one example of many embodiments described herein, the contact with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors is made 3, 4, 5, 6, 7, 8, 9, or 10 days after the application of the alternating current electric field.

[0181] In one example of many embodiments described herein, the contact is made at a time when the response of the subject to the alternating current electric field has decreased.

[0182] In one example of many embodiments described herein, the contact is made 3, 4, 5, 6, 7, 8, 9, or 10 days after step a) is performed.

[0183] In one example of many embodiments described herein, the alternating current electric field is applied before, after, or simultaneously with the administration of one or more FGF or FGFR inhibitors.

[0184] In one example of many embodiments described herein, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (targeted or non-targeted), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, intraoperative, or during biopsy), or by inhalation.

[0185] In one example of many embodiments described herein, the FGF inhibitor inhibits or decreases FGF expression.

[0186] In one example of many embodiments described herein, the FGF inhibitor blocks the upregulation of FGF expression in response to an alternating electric field.

[0187] In one example of many embodiments described herein, the FGF inhibitor prevents FGF from interacting or binding with one or more fibroblast growth factor receptors (FGFRs).

[0188] In one example of many embodiments described herein, the FGFR inhibitor prevents FGFR from interacting or binding with one or more FGFs.

[0189] In one example of many embodiments described herein, the FGFR inhibitor inhibits one or more fibroblast growth factor receptors (FGFRs).

[0190] In one example of many embodiments described herein, the FGF is FGF-21, FGF-19, FGF-7, or FGF-basic.

[0191] In one example of many embodiments described herein, the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

[0192] In one example of many embodiments described herein, the FGFR inhibitor can be one or more of the inhibitors in Table 1.

[0193] In one example of many embodiments described herein, the frequency of the alternating electric field is 50 kHz to 1 MHz.

[0194] In one example of many embodiments described herein, the frequency of the alternating electric field is 150 kHz or 250 kHz.

[0195] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.5 to 4 V / cm RMS.

[0196] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.9 V / cm RMS.

[0197] In one example of many embodiments described herein, the method further includes the step of administering a cancer therapeutic agent.

[0198] One example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in the treatment of a subject in need thereof.

[0199] In one example of many embodiments described herein, the alternating electric field is applied at a constant frequency to a target site of a subject in need thereof for a certain period of time.

[0200] In one example of many embodiments described herein, the subject has a mesothelioma, ovarian cancer, or lung cancer.

[0201] In one example of many embodiments described herein, the target site includes one or more cancer cells.

[0202] In one example of many embodiments described herein, the alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF or FGFR inhibitors.

[0203] In one example of many embodiments described herein, the one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (target or non-target), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, intraoperative, or during biopsy), or by inhalation.

[0204] An example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in enhancing the sensitivity of cells to an alternating electric field.

[0205] An example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in enhancing the sensitivity of cells within a subject to an alternating electric field.

[0206] An example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in enhancing the sensitivity of cells to an alternating electric field, wherein the method is performed in vitro on the cells.

[0207] An example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in enhancing the cytotoxicity within cells.

[0208] An example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in enhancing the cytotoxicity within cells within a subject.

[0209] An example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in enhancing the cytotoxicity within cells, wherein the method is performed in vitro on the cells.

[0210] In one example of many embodiments described herein, an alternating electric field is applied to cells at a constant frequency for a certain period of time.

[0211] In one example of many embodiments described herein, one or more FGF inhibitors or FGFR inhibitors are contacted with cells.

[0212] In one example of many embodiments described herein, an alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF or FGFR inhibitors.

[0213] In one example of many embodiments described herein, in the foregoing embodiments, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (target or non-target), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, during surgery or biopsy), or by inhalation.

[0214] In one example of many embodiments described herein, the cells are in vitro.

[0215] In one example of many embodiments described herein, the cells are within a subject.

[0216] In one example of many embodiments described herein, the cancer cells are ovarian cancer or lung cancer cells.

[0217] One example of many embodiments described herein is a combination of an alternating electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in maintaining the sensitivity of cells to the alternating electric field.

[0218] An example of many embodiments described herein is a combination of an alternating current electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in maintaining the sensitivity of cells in a subject to an alternating current electric field.

[0219] An example of many embodiments described herein is a combination of an alternating current electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in maintaining the sensitivity of cells to an alternating current electric field, the method being the combination performed on the cells in vitro.

[0220] An example of many embodiments described herein is a combination of an alternating current electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in maintaining the sensitivity of a subject to an alternating current electric field.

[0221] An example of many embodiments described herein is a combination of an alternating current electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in maintaining the sensitivity of a subject to an alternating current electric field when the cells are within the subject.

[0222] An example of many embodiments described herein is a combination of an alternating current electric field and one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors for use in maintaining the sensitivity of a subject to an alternating current electric field, the method being the combination performed on the cells in vitro.

[0223] In an example of many embodiments described herein, FGF expression is detected in a subject or in cells.

[0224] In one example of many embodiments described herein, the response of a cell or subject to an alternating electric field decreases before using one or more FGF inhibitors or FGFR inhibitors.

[0225] In one example of many embodiments described herein, the use of one or more FGF inhibitors or FGFR inhibitors is performed 3, 4, 5, 6, 7, 8, 9, or 10 days after applying an alternating electric field.

[0226] In one example of many embodiments described herein, an alternating electric field is applied before, after, or simultaneously with administering one or more FGF or FGFR inhibitors.

[0227] In one example of many embodiments described herein, one or more FGF or FGFR inhibitors are administered intratumorally, intracranially, intraventricularly, intrathecally, epidurally, intradurally, intravascularly, intravenously (targeted or non-targeted), intraarterially, intramuscularly, subcutaneously, intraperitoneally, orally, intranasally, by intratumoral injection (e.g., computer tomography-guided, during surgery or biopsy), or by inhalation.

[0228] In one example of many embodiments described herein, an FGF inhibitor inhibits or decreases FGF expression.

[0229] In one example of many embodiments described herein, an FGF inhibitor blocks the upregulation of FGF expression in response to an alternating electric field.

[0230] In one example of many embodiments described herein, an FGF inhibitor prevents FGF from interacting or binding with one or more fibroblast growth factor receptors (FGFRs).

[0231] In one example of many embodiments described herein, an FGFR inhibitor prevents FGFR from interacting or binding with one or more FGFs.

[0232] In one example of many embodiments described herein, the FGFR inhibitor blocks one or more fibroblast growth factor receptors (FGFRs).

[0233] In one example of many embodiments described herein, the FGF is FGF-21, FGF-19, FGF-7, or FGF-basic.

[0234] In one example of many embodiments described herein, the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

[0235] In one example of many embodiments described herein, the FGFR inhibitor can be one or more of the inhibitors in Table 1.

[0236] In one example of many embodiments described herein, the frequency of the alternating electric field is 50 kHz to 1 MHz.

[0237] In one example of many embodiments described herein, the frequency of the alternating electric field is 50 kHz or 250 kHz.

[0238] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.5 to 4 V / cm RMS.

[0239] In one example of many embodiments described herein, the alternating electric field has an electric field strength of 0.9 V / cm RMS.

[0240] One of ordinary skill in the art will recognize many, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the methods and compositions described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

1. A pharmaceutical composition for use in a method of treating a subject in need, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising the steps of: applying the alternating electric field at a constant frequency to a target site of the subject in need for a certain period of time; and administering the fibroblast growth factor (FGF) inhibitor or the fibroblast growth factor receptor (FGFR) inhibitor to the subject in need.

2. A pharmaceutical composition for use in a method to enhance the sensitivity of cells to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising the steps of: applying the alternating electric field to cells at a constant frequency for a certain period of time; and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating electric field.

3. A pharmaceutical composition for use in a method to enhance the sensitivity of cells to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, wherein the method comprises the steps of: applying the alternating electric field to cells at a constant frequency for a certain period of time; and contacting the cells with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cells to the alternating electric field, wherein the cells are located within the subject.

4. A pharmaceutical composition for use in a method to enhance the sensitivity of a cell to an alternating electric field, comprising a factor receptor (FGFR) inhibitor, the method comprising the steps of applying the alternating electric field to the cell at a constant frequency for a certain period of time, and contacting the cell with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing the sensitivity of the cell to an alternating electric field, the method being performed on the cell in vitro, the pharmaceutical composition.

5. A pharmaceutical composition for use in a method to enhance intracellular cytotoxicity, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, wherein the method comprises the steps of: applying an alternating electric field at a constant frequency to a cell for a certain period of time; and contacting the cell with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing intracellular cytotoxicity, wherein the cell is located within a subject.

6. A pharmaceutical composition for use in a method to enhance intracellular cytotoxicity, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising the steps of: applying an alternating electric field at a constant frequency to a cell for a certain period of time; and contacting the cell with one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors, thereby enhancing intracellular cytotoxicity, the method being performed in vitro on a cell, the pharmaceutical composition.

7. The subject is alive, the pharmaceutical composition according to any one of claims 1 to 6.

8. The pharmaceutical composition according to any one of claims 1 to 6, wherein the subject is suffering from mesothelioma, ovarian cancer, or lung cancer.

9. The pharmaceutical composition according to any one of claims 1 to 6, wherein the target site comprises one or more cancer cells.

10. The pharmaceutical composition according to any one of claims 1 to 6, wherein the alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF inhibitors or FGFR inhibitors.

11. The pharmaceutical composition according to any one of claims 1 to 6, wherein the one or more FGF inhibitors or FGFR inhibitors are administered intratumor, intracranial, intraventricular, intrathecal, epidural, intradural, intravascular, intravenous (targeted or non-targeted), intraarterial, intramuscular, subcutaneous, intraperitoneal, oral, intranasal, intratumor injection, or inhalation.

12. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGF inhibitor inhibits or reduces FGF expression.

13. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGF inhibitor blocks the upregulation of FGF expression in response to an alternating electric field.

14. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGF inhibitor prevents FGF from interacting with or binding to one or more FGF receptors (FGFRs).

15. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGFR inhibitor prevents FGFR from interacting with or binding to one or more FGFs.

16. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGFR inhibitor blocks one or more FGF receptors (FGFRs).

17. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGF is FGF-21, FGF-19, FGF-7, or FGF-basic.

18. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

19. The pharmaceutical composition according to any one of claims 1 to 6, wherein the FGFR inhibitor may be one or more of the inhibitors listed in Table 1. Table 1 Table 1

20. The pharmaceutical composition according to claim 9, wherein the cancer cells are ovarian cancer cells or lung cancer cells.

21. The pharmaceutical composition according to any one of claims 1 to 6, wherein the frequency of the alternating electric field is 50 kHz to 1 MHz.

22. The pharmaceutical composition according to any one of claims 1 to 6, wherein the frequency of the alternating electric field is 150 or 250 kHz.

23. The pharmaceutical composition according to any one of claims 1 to 6, wherein the electric field strength of the alternating electric field is 0.5 to 4 V / cm RMS.

24. The pharmaceutical composition according to any one of claims 1 to 6, wherein the electric field strength of the alternating electric field is 0.9 V / cm RMS.

25. The pharmaceutical composition according to any one of claims 1 to 6, further comprising the step of administering a cancer drug.

26. A pharmaceutical composition for use in a method for maintaining the sensitivity of cells to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising the steps of applying the alternating electric field to cells at a constant frequency for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating electric field, according to any one of claims 1 to 6.

27. ​​A pharmaceutical composition for use in a method for maintaining the sensitivity of cells to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, wherein the method comprises the steps of applying the alternating electric field to cells at a constant frequency for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating electric field within the cells, wherein the cells are located within a subject, according to any one of claims 1 to 6.

28. A pharmaceutical composition for use in a method for maintaining the sensitivity of cells to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising the steps of applying an alternating electric field to cells at a constant frequency for a certain period of time, and contacting the cells with one or more FGF inhibitors or FGFR inhibitors, thereby maintaining the sensitivity of the cells to the alternating electric field, the method being performed in vitro on the cells, the pharmaceutical composition according to any one of claims 1 to 6.

29. A pharmaceutical composition for use in a method for maintaining a subject's sensitivity to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising applying an alternating electric field at a constant frequency to a target site of the subject for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to the subject in need, thereby maintaining the sensitivity of the cells to the alternating electric field.

30. A pharmaceutical composition for use in a method for maintaining the sensitivity of cells to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising applying an alternating electric field at a constant frequency to a target site of a subject in need for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to the subject in need, thereby maintaining the sensitivity of cells to an alternating electric field.

31. A pharmaceutical composition for use in a method for maintaining a subject's sensitivity to an alternating electric field, comprising a fibroblast growth factor (FGF) inhibitor or a fibroblast growth factor receptor (FGFR) inhibitor, the method comprising applying an alternating electric field at a constant frequency to a target site of the subject for a certain period of time, and administering one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to the subject in need, thereby maintaining the subject's or cell's sensitivity to an alternating electric field.

32. The pharmaceutical composition according to any one of claims 1 to 6 and 29 to 31, further comprising the step of detecting an increase in FGF expression in the subject or the cells before administering the one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors to the subject in need, after applying the alternating electric field.

33. The pharmaceutical composition according to any one of claims 1 to 6 and 29 to 31, wherein the contact is made when the cell's response to an alternating electric field decreases.

34. The pharmaceutical composition according to any one of claims 1 to 6 and 29 to 31, wherein contact with the one or more fibroblast growth factor (FGF) inhibitors or fibroblast growth factor receptor (FGFR) inhibitors is performed 3, 4, 5, 6, 7, 8, 9, or 10 days after the application of the alternating electric field.

35. The pharmaceutical composition according to any one of claims 1 to 6 and 29 to 31, wherein the contact is made when the subject's response to the alternating electric field decreases.

36. The pharmaceutical composition according to any one of claims 1 to 6 and 29 to 31, wherein the contact is performed 3, 4, 5, 6, 7, 8, 9, or 10 days after step a).

37. The pharmaceutical composition according to any one of claims 29 to 31, wherein the alternating electric field is applied before, after, or simultaneously with the administration of one or more FGF inhibitors or FGFR inhibitors.

38. The pharmaceutical composition according to any one of claims 29 to 31, wherein the one or more FGF inhibitors or FGFR inhibitors are administered intratumor, intracranial, intraventricular, intrathecal, epidural, intradural, intravascular, intravenous (targeted or non-targeted), intraarterial, intramuscular, subcutaneous, intraperitoneal, oral, intranasal, intratumor injection, or inhalation.

39. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGF inhibitor inhibits or reduces FGF expression.

40. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGF inhibitor blocks the upregulation of FGF expression in response to an alternating electric field.

41. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGF inhibitor prevents FGF from interacting with or binding to one or more FGF receptors (FGFRs).

42. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGFR inhibitor prevents FGFR from interacting with or binding to one or more FGFs.

43. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGFR inhibitor blocks one or more FGF receptors (FGFRs).

44. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGF is FGF-21, FGF-19, FGF-7, or FGF-basic.

45. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGFR is FGFR1, FGFR2, FGFR3, or FGFR4.

46. The pharmaceutical composition according to any one of claims 29 to 31, wherein the FGFR inhibitor may be one or more of the inhibitors in Table 1. Table 1 Table 2

47. The pharmaceutical composition according to any one of claims 29 to 31, wherein the frequency of the alternating electric field is 50 kHz to 1 MHz.

48. The pharmaceutical composition according to any one of claims 29 to 31, wherein the frequency of the alternating electric field is 150 or 250 kHz.

49. A pharmaceutical composition as described in any one of the items.

50. The pharmaceutical composition according to any one of claims 29 to 31, wherein the electric field strength of the alternating electric field is 0.9 V / cm RMS.

51. The pharmaceutical composition according to any one of claims 29 to 31, further comprising the step of administering a cancer drug.

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