Non-invasive cancer treatment

Abstract

An apparatus 1 for treating a cancerous target site is provided. The apparatus comprises an electromagnetic emitter 10 including one or more electrodes having an electrically insulating coating that prevents electrical contact between the electrodes and the target site. The electromagnetic emitter 10 is configured to provide a tumor treatment region at the target site 30 via the one or more electrodes, the tumor treatment region being a non-ionizing alternating electromagnetic field having a frequency between 10 kHz and 300 kHz and further having a magnetic flux density between 0.1 pT and 1 mT. The apparatus further comprises a heat source 20 configured to provide heating at the target site and induce hyperthermia at the target site. The apparatus is configured to apply the non-ionizing alternating electromagnetic field and the heating independently. The apparatus comprises an electronic controller for electronically controlling the electromagnetic emitter and the heat source.

Description

[Technical Field]

[0001] The present disclosure relates to devices for treating cancerous target sites. More specifically, the present disclosure relates to devices configured to deliver a non-ionizing alternating electromagnetic field and localized heating to a target site. The present disclosure also relates to related methods of treating cancerous sites and compositions used to treat cancerous target sites. [Background technology]

[0002] Non-invasive methods for treating cancer are important, especially for cancers such as glioblastoma that are difficult to remove by surgery. New non-invasive methods for treating cancer that effectively reduce or eliminate cancerous cells from target sites in the body are desirable.

[0003] Radiation therapy is a type of cancer therapy that uses ionizing radiation to kill malignant cells at a target site. The ionizing radiation is delivered to the target site and causes damage through the direct or indirect action of the radiation on DNA and other cellular molecules. In direct action, the radiation directly strikes, for example, DNA molecules, disrupting their molecular structure. Such structural changes cause cell damage or even cell death, thereby providing a mechanism for treating malignant cells.

[0004] In another form of treatment, known as alternating current electric field therapy or tumor treating field (TT field), a non-ionizing electric field is applied to the target site. The mechanism that makes TT field useful for cancer treatment differs from that of radiation therapy. Specifically, microtubule assemblies are distorted during the formation of the mitotic spindle. Tumor cells undergo mitosis and remain in interphase for extended periods of time. When a cleavage furrow forms during metaphase or anaphase, all polar molecules and dipoles within the cell undergo dielectrophoresis under the influence of the TT field, accumulating within the cleavage furrow and ultimately rupturing the cell membrane. Mitotic consequences induced by application of the TT field include abnormal chromosome segregation, which leads to various forms of cell death.

[0005] Hyperthermia is a known method for treating cancerous tissue. However, effectively targeting the cancerous tissue can be difficult. On the one hand, tissue surrounding the treated area may also be affected by hyperthermia, especially if the cancerous tissue is heated to a high temperature. On the other hand, the cancerous tissue may not be effectively destroyed if the tissue is not heated to a sufficiently high temperature. Furthermore, while applying heat to a cancerous tissue can cause cell damage or even cell death at the cancerous tissue, the cancerous tissue may resist such heating if the heat is not applied properly and accurately to the cancerous tissue in the appropriate amount. For example, if hyperthermia heats healthy surrounding tissue in addition to the cancerous growth, increased blood flow may increase the supply of nutrients to the cancerous growth. Therefore, in some situations, increased blood flow as a result of heating may promote the dispersion of nutrients, allowing them to reach the cancerous site, resulting in the opposite effect to the intended treatment, or at least ineffective treatment. Furthermore, it has been observed that different stages in the cell cycle of cancerous cells have different resistance to heating. Furthermore, elevated temperatures in cells transiently upregulate heat shock genes encoding heat shock proteins (HSPs). The mechanism involved in the heat shock response is an autoregulatory loop. HSPs normally maintain the associated transcription factor (HSF-1) in an inactive state, but heating induces HSPs to bind unfolded proteins with higher affinity, triggering the release of HSF-1 from the HSPs and initiating HSP gene transcription. When protein damage / aggregation is repaired after heat shock by HSPs, substrate-free HSPs themselves may be involved in alleviating the response by rebinding to HSF-1. As a result, HSP levels transiently increase after heating but gradually decrease again after a prolonged period of stress-free survival. HSP upregulation is closely associated with a transient state of resistance of cells to a subsequent second heat shock. Increased HSP levels are thought to protect cells from further heating-induced protein damage through their chaperone activity.Therefore, there is a need to provide hyperthermia that more accurately and reliably delivers the required heating to the cancerous site to avoid resistance of cancerous cells to heating as a result of deviations from the required applied heat dose.

[0006] Many methods of hyperthermia require the administration of a mediator (such as a nanoparticle fluid) to the site. The mediator is then heated, for example, by an electric field, thereby indirectly heating the target site. The mediator can be heated using an electric field having the same or similar frequency range as the TT region. This can be considered advantageous because the cancerous site is attacked by both the hyperthermia and TT region mechanisms. However, because the mediator is a fluid injected into the patient's body, it is difficult to control the location of heating when administering electromagnetic radiation. Summary of the Invention [Problem to be solved by the invention]

[0007] It is desirable to find more effective cancer treatment methods than those described above. [Means for solving the problem]

[0008] The present invention provides an apparatus for treating a cancerous target site, comprising: an electromagnetic emitter configured to provide a non-ionizing alternating electromagnetic field at a target site; a heat source configured to provide heating to the target site to induce hyperthermia at the target site; The present invention addresses the problem of providing a more effective cancer treatment method by providing an apparatus configured to independently apply a non-ionizing alternating electromagnetic field and heat, comprising: a heat source configured to provide heating directly to the target site;

[0009] It should be noted herein that a heat source can be any heat source that heats a specific target site without the use of a mediator (e.g., directly, rather than indirectly via a mediator).

[0010] In a preferred embodiment, the device is configured to deliver a non-ionizing alternating electromagnetic field to the target for a first time period and deliver direct heating to the target site for a second time period. The second time period may partially or completely overlap with the first time period. For example, the first time period may begin simultaneously with the second time period, or may begin a predetermined time after the start of the first time period, or may begin when the first time period ends. Preferably, the first and second times completely overlap, so that the TT field and heating are applied to the site simultaneously, resulting in a synergistic effect on cells within the target site, as discussed in more detail below.

[0011] It should be noted that the sequence of first and second periods may be repeated two or more times. For example, the device may be configured to apply a non-ionizing alternating electromagnetic field for a first period of time and apply heating for a second period of time a predetermined time after the start of the first period of time. Subsequently, the device may again be configured to apply a non-ionizing alternating electromagnetic field for the first period of time and apply heating for a second period of time a predetermined time after the start of the first period of time. The lengths of the first and second periods of time and their relative start times may be configurable by a user or according to one or more schedules stored in the memory of the device.

[0012] The device is preferably configured to deliver a non-ionizing alternating electromagnetic field to the target site for a first period of time ranging from 1 minute to 24 hours.

[0013] In another preferred embodiment, the device is further configured to apply heating to the target site for a second period of time ranging from 1 minute to 360 minutes. Heating can be applied simultaneously with the non-ionizing alternating electromagnetic field for a third period of time. The third period of time can be all or a portion of the second period of time.

[0014] In another preferred embodiment, the electromagnetic emitter is configured to provide an alternating electromagnetic field having a frequency of 10 kHz to 500 kHz, more preferably 10 kHz to 300 kHz, and even more preferably 100 kHz to 300 kHz. A significant increase in the therapeutic effect of the tumor treatment region has been observed when the frequency is less than 300 kHz.

[0015] In another preferred embodiment, the electromagnetic emitter is configured to provide an alternating electromagnetic field at the target site having a magnetic flux density of 0.1 pT to 1 mT, or 0.1 pT to 100 μT, or 100 μT to 1 mT and / or an electric field strength of 1 V / cm to 3 V / cm.

[0016] In another preferred embodiment, the electromagnetic emitter is configured to provide an alternating electromagnetic field at the target site having a magnetic flux density between 0.5 μT and 1 mT, more preferably between 8 μT and 1 mT.

[0017] In another preferred embodiment, the heat source is configured to heat the target site to a temperature of at least 42° C., preferably between 42° C. and 57° C. Heating the target site to a temperature of at least 42° C. induces a heating effect at the target site equivalent to extreme hyperpyrexia.

[0018] In another preferred embodiment, the heat source comprises an ultrasound emitter configured to provide ultrasound radiation to the target site, optionally with one or more focal regions at the target site, which may be provided by a single transducer or by multiple transducers.

[0019] In a further preferred embodiment, the heat source comprises: an electromagnetic emitter configured to provide electromagnetic radiation to a target site; a fluid pump configured to pump the fluid to the target site and a heater to heat the fluid before it reaches the target site; and / or a conductive heat emitter configured to supply heat to the target site by thermal conduction; It has one or more of the following.

[0020] In a further preferred embodiment, the device additionally comprises an electronic control unit for electronically controlling the electromagnetic emitter and the heat source.

[0021] A further aspect of the present invention relates to a method of treating a cancerous target site using a device according to the above-described embodiments, where the device can have any of the configurations disclosed herein, and the method is preferably carried out by appropriately positioning the emitter 10 and heat source 20 on a patient. For example, an applicator of the electromagnetic emitter 10 can be placed at a predetermined location on the patient's body, and the heat source 20 (e.g., the transducer 204 described in connection with FIG. 2) can also be appropriately positioned. Here, the method includes:

[0022] In step S1, an electromagnetic emitter 10 is used to provide a non-ionizing alternating electromagnetic field, generating a magnetic flux density at the target site.

[0023] In step S2, heat (more specifically, direct heat) is applied to the target site using heat source 20. This can be applied simultaneously with, or a predetermined time before or after, applying the electromagnetic field to target site 30. In some embodiments, an alternating electromagnetic field alone may be applied without heating for a first period of time to create a magnetic flux density at the target site, and then both may be applied to the target site simultaneously.

[0024] Optionally, in step S3, an anti-tumorigenic composition is delivered to the target site. The anti-tumorigenic composition can be administered by any suitable means, such as orally or intravenously. It should be noted that the anti-tumorigenic composition can be delivered before or simultaneously with steps S1, S2, and S4, or a predetermined time after step S1, S2, or S4.

[0025] Examples of anti-tumorigenic compositions that can be used to practice the present invention are detailed throughout the present invention.

[0026] Optionally, in step S4, a glutathione (GSH)-depleted composition is provided to the target site in addition to the anti-tumorigenic compound. The GSH-depleted composition can be administered by any suitable means, such as orally or intravenously. It should be noted that the GSH-depleted composition can be provided before or simultaneously with steps S1, S2, and S3, or a predetermined time after step S1, S2, or S3, preferably a predetermined time after step S3. Examples of GSH-depleted compositions that can be used to practice the present invention are described in detail throughout the present invention.

[0027] Direct heating is stopped in step S5, and the non-ionizing AC electric field is stopped in step S6. Note that steps S5 and S6 may be performed simultaneously, or the non-ionizing AC electric field may be stopped after direct heating is stopped, such that only the non-ionizing AC electric field is applied for a predetermined period of time after direct heating is stopped.

[0028] A further aspect of the present invention relates to an anti-tumorigenic composition for use in a method of treating a cancerous target site using a device according to the above-described embodiments, wherein the device can have any of the configurations disclosed herein, and the method is preferably carried out by appropriately positioning the emitter 10 and heat source 20 on a patient. For example, an applicator of the electromagnetic emitter 10 can be placed at a predetermined location on the patient's body, and the heat source 20 (e.g., transducer 204) can also be appropriately positioned. wherein the method includes:

[0029] In step S1, an electromagnetic emitter 10 is used to provide a non-ionizing alternating electromagnetic field, generating a magnetic flux density at the target site.

[0030] In step S2, heat (more specifically, direct heat) is applied to the target site using heat source 20. This can be applied simultaneously with, or a predetermined time before or after, applying the electromagnetic field to target site 30. In some embodiments, an alternating electromagnetic field alone may be applied without heating for a first period of time to create a magnetic flux density at the target site, and then both may be applied to the target site simultaneously.

[0031] In step S3, the anti-tumorigenic composition is delivered to the target site. The anti-tumorigenic composition can be administered by any suitable means, such as orally or intravenously. It should be noted that the anti-tumorigenic composition can be delivered before or simultaneously with steps S1, S2, and S4, or a predetermined time after step S1, S2, or S4. Examples of anti-tumorigenic compositions that can be used to practice the present invention are described in detail throughout the present invention.

[0032] Optionally, in step S4, a glutathione (GSH)-depleted composition is provided to the target site in addition to the anti-tumorigenic compound. The GSH-depleted composition can be administered by any suitable means, such as orally or intravenously. It should be noted that the GSH-depleted composition can be provided before or simultaneously with steps S1, S2, and S3, or a predetermined time after step S1, S2, or S3, preferably a predetermined time after step S3. Examples of GSH-depleted compositions that can be used to practice the present invention are described in detail throughout the present invention.

[0033] Direct heating is stopped in step S5, and the non-ionizing AC electric field is stopped in step S6. Note that steps S5 and S6 may be performed simultaneously, or the non-ionizing AC electric field may be stopped after direct heating is stopped, such that only the non-ionizing AC electric field is applied for a predetermined period of time after direct heating is stopped.

[0034] Embodiments will now be described in detail, by way of non-limiting example only, with reference to the accompanying drawings, in which: [Brief explanation of the drawings]

[0035] [Figure 1] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more embodiments. [Figure 2] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 3] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 4] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 5] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 6] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 7] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 8] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 9] FIG. 1 is a schematic diagram of an apparatus for treating a cancerous target site according to one or more further embodiments. [Figure 10] FIG. 1 illustrates an overview of an apparatus for treating a cancerous target site according to one or more embodiments. [Figure 11] FIG. 1 illustrates an overview of a method for treating a cancerous target site using a device according to the present disclosure. [Figure 12]Figure 1 shows experimental data demonstrating the in vitro effects of TT domain ("TTF"), hyperthermia ("HT"), pterostilbene ("PT"), and their combination on U87MG cells. The TT domain was applied at 300 kHz for 240 minutes (from 0 to 240 minutes) at an average magnetic flux density of 8 μT. The hyperthermia was applied at 42°C for 10 minutes (from 120 to 130 minutes). 20 μM pterostilbene was applied for 120 minutes (from 120 to 240 minutes). Data represent the average number of viable cells for five experiments, with * vs. control, + vs. TTF, and # vs. TTF + HT, P<0.01 using Student's t-test. [Figure 13] FIG. 1 shows in vitro experimental data on the effect of exposure to an oscillating magnetic field on cell viability of U87MG (ATCC) cells. [Figure 14] FIG. 1 shows in vitro experimental data on the effect of heat exposure on cell viability of U87MG cells. [Figures 15A-15F] FIG. 1 shows microscopic and control images of in vitro U87MG cell cultures after exposure to different external treatments. [Figure 16] FIG. 1 shows in vitro experimental data on the effects of exposure to electromagnetic fields, heat, and temozolomide (TMZ) on cell viability of U87MG cells. [Figure 17] FIG. 1 shows in vitro experimental data on the effects of exposure to electromagnetic fields, heat, and resveratrol or its derivatives on cell viability of U87MG cells. [Figure 18] FIG. 1 shows in vitro experimental data on the effect of exposure to an oscillating magnetic field on cell viability of AsPC1 (pancreatic adenocarcinoma, ATCC) cells. [Figure 19] FIG. 1 shows in vitro experimental data on the effect of heat exposure on cell viability of AsPC1 cells. [Figure 20]FIG. 1 shows experimental data showing the in vitro effects of TT-domain ("TTF"), hyperthermia ("HT"), pterostilbene ("PT") and their combinations on AsPC1 cells. [Figures 21A-21H] FIG. 1 shows microscopic and control images of in vitro AsPC1 cell cultures after exposure to different external treatments. [Figure 22] FIG. 1 shows experimental data showing the in vitro effects of TT-field ("TTF"), hyperthermia ("HT"), gemcitabine ("GEM"), pterostilbene ("PT") and their combinations on AsPC1 cells. [Figure 23] FIG. 1 shows in vitro experimental data on the effect on cell viability of A2058 (melanoma, ATCC) cells upon exposure to an oscillating magnetic field. [Figure 24] FIG. 1 shows in vitro experimental data on the effect of heat exposure on cell viability of A2058 cells. [Figure 25] FIG. 1 shows experimental data showing the in vitro effects of TT-domain ("TTF"), hyperthermia ("HT"), pterostilbene ("PT") and their combinations on A2058 cells. [Figures 26A-26H] FIG. 1 shows microscopic and control images of in vitro A2058 cell cultures after exposure to different external treatments. [Figure 27] FIG. 1 shows experimental data showing the in vitro effects of TT domain ("TTF" and "TTF" are used interchangeably), hyperthermia ("HT"), paclitaxel ("PAC") and their combinations on A2058 cells. DETAILED DESCRIPTION OF THE INVENTION

[0036] The present invention relates to a device configured to deliver both a non-ionizing alternating current electromagnetic field and hyperthermia to a target site, and the electromagnetic field and hyperthermia can be delivered independently. The electromagnetic field can be applied, for example, by a magnetic applicator that delivers magnetic flux density to the target site. Hyperthermia treatments using a mediator deliver an alternating current electromagnetic field that may be absorbed by the mediator. However, because the mediator absorbs electromagnetic energy, the tumor therapeutic efficacy of the electromagnetic field can be reduced. The present invention overcomes this problem by delivering hyperthermia independently, allowing both treatments to be delivered to a cancerous site without reducing the effectiveness of the other. In the present invention, the tumor therapeutic effect is due to the combined use of the electromagnetic field and direct hyperthermia. The electromagnetic field can be delivered using a magnetic field applicator. In some further examples of the present disclosure, the treatment can further include administration of an anti-cancer composition.

[0037] As used herein, the terms "tumor treatment region," "TT region," or "TTF" may be understood to mean an oscillating electromagnetic field applied to a target site. In particular, the electromagnetic field is generated by an applicator that is electrically isolated from the target site so that no current flows between the target site and the applicator.

[0038] FIG. 1 shows a schematic diagram of an apparatus 1 for treating a cancerous target site 30. The apparatus includes an electromagnetic emitter 10 and a heat source 20. The electromagnetic emitter 10 is configured to provide a non-ionizing alternating electromagnetic field 15 to the target site 30. The heat source 20 is configured to provide direct heating 25 to the target site 30, inducing hyperthermia at the target site 30. In one configuration, the apparatus 1 is configured to provide the non-ionizing alternating electromagnetic field and direct heating to the target site 30 within a predetermined time period. The target site 30 may include at least a cancerous growth and may further include a portion of tissue surrounding the cancerous growth. In some embodiments, the apparatus 1 may be configured to provide the non-ionizing alternating magnetic field 15.

[0039] As disclosed herein, hyperthermia can be defined as an increase in temperature above 37.5°C. Accordingly, the devices disclosed herein can be configured to heat a target site to temperatures above 37.5°C. Note that hyperthermia is defined as a temperature above 37.5°C to 38.3°C (depending on the criteria used) that occurs without a change in the body temperature set point. In contrast, hyperthermia is an extreme increase in body temperature and is classified as a core body temperature above 40.0°C or 41.0°C, depending on its origin. The range of hyperthermia includes cases considered severe (above 40°C) and extreme (above 42°C). Hyperthermia differs from hyperthermia in that the body temperature set point of an individual's thermoregulatory system is set higher than normal, resulting in the body generating heat to achieve that temperature set point. In contrast, hyperthermia involves an external factor causing an increase in body temperature above the set point.

[0040] Additionally, it is noted that thermal ablation is a type of procedure that uses heat, cold, microwaves, and electrical current to vaporize (ablate) cancer cells and tumors by heating them to above 50°C.

[0041] In a preferred embodiment, the device is configured to heat the target site to a temperature of 39°C to 52°C (heating above 39°C can sensitize cancerous growths to other therapies such as TT regions, chemotherapy, and radiation therapy), preferably at least 41.1°C (above which advantageously causes irreversible damage to the cells). The heat source is preferably configured to heat the target site to a temperature of at least 42°C, preferably 42°C to 57°C. Heating the target site to a temperature of at least 42°C induces a heating effect at the target site equivalent to extreme hyperthermia.

[0042] Heat source 30 can be any suitable heat source that provides direct heating to target site 30, and can include electromagnetic heating, such as capacitive radio frequency heating, radiative radio frequency heating, microwave heating, infrared heating, and laser heating, ultrasonic heating, heating via a heated fluid, heating via a conductive heat emitter, or any other suitable method of heating target site 30 independent of non-ionizing alternating electromagnetic field 15. Heat source 30 can be any heat source that heats the target site without the use of a mediator (e.g., directly rather than indirectly via a mediator).

[0043] In any of the embodiments disclosed herein, the electromagnetic emitter 10 may be configured to produce an electromagnetic field 15 having a frequency between 10 kHz and 500 kHz, more preferably between 10 kHz and 300 kHz. The electromagnetic field may have a magnetic flux density between 0.1 pT and 1 mT, or between 0.1 pT and 100 μT, or between 100 μT and 1 mT, and / or a corresponding electric field between 1 V / cm and 3 V / cm, depending on tissue impedance (i.e., taking into account the attenuation of the electric field that may occur as it travels from the electromagnetic emitter 10 to the target site 30, which can be determined from the impedances created by different types of tissue present between the electromagnetic emitter 10 and the target site 30). As noted above, the electromagnetic field 15 is non-ionizing, and its mechanism of action on cancerous sites differs from that of ionizing radiation, as discussed in the Background section of this disclosure. Furthermore, the electromagnetic field 15 itself does not directly heat the target site 30 due to the relatively low strength of the oscillating field.

[0044] It will be understood that in any of the embodiments disclosed herein, the electromagnetic emitter 10 and heat source 20 can be powered by any power source, and can be powered by the same or different power sources. Similarly, the electromagnetic emitter 10 and heat source 20 can each include a user interface for selecting the operating parameters (frequency, field strength, amplitude, etc.) of each emitter, or the emitter 10 and heat source 20 can include pre-programmed sequences for emitting electromagnetic radiation and heat according to a user-selectable predetermined program.

[0045] In any of the embodiments disclosed herein, the device may further include a temperature measurement element for measuring the temperature of the target site 30. For example, the device may include an implantable temperature measurement probe configured to be implanted proximate the target site 30 to measure a temperature indicative of the temperature of the target site 30. The probe may include, for example, a thermocouple, a thermistor, and / or a fiber optic sensor. In other embodiments, non-invasive temperature measurements such as infrared sensing, CT thermometry, or magnetic resonance thermometry may be used.

[0046] FIG. 2 shows a schematic diagram of an electromagnetic emitter 10 and a heat source 200 according to one or more embodiments.

[0047] The electromagnetic emitter 10 may include one or more sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be placed on or proximate to the surface of a patient's body 35 and are electrically isolated from the patient's body (i.e., do not form a closed electrical circuit between the source 12 and the patient's body). In some embodiments, the applicators 14 may include one or more electrodes having an electrically insulating coating that prevents electrical contact between the electrodes and the patient's surface and, therefore, the target site. Note that even when the applicators are placed on the patient's surface of the patient's body 35, they remain electrically isolated from the body, e.g., by the presence of the electrically insulating coating. The source 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward the target site 30 and provides a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the magnetic flux density produced at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14. In some embodiments, the applicator 14 may include a coil having positive and negative terminals. One or more sources may be configured to provide alternating current through the coil to generate a magnetic field from the coil. The coil may include any number of turns, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 100, 500, or more.

[0048] The heat source 200 shown in FIG. 2 may be an ultrasound emitter and may include an ultrasound source 202 that emits an ultrasound signal to one or more transducers 204. The transducers 204 are configured to transmit focused ultrasound radiation 205 to the target region 30. The one or more transducers 204 may include, for example, one or more piezoelectric transducers. The one or more transducers 204 may include one or more plastic and / or ceramic transducers. To improve the transmission of ultrasound waves from the transducers 204 to the interior of the patient's body 30 (i.e., to reduce reflections of ultrasound waves), a couplant (not shown) may be applied on the patient's body 35 between the transducers 204 and the patient's body 30. Couplant is defined herein as any suitable solid or liquid (or combination thereof) for improving the transmission of ultrasound waves from the transducers 204 to the interior of the patient's body 30. The shape of the transducer 204 can be selected to select the amount of ultrasound focusing, and can be selected to focus the ultrasound radiation 205 at one or more focal regions within the target site 30. For example, one or more transducers 204 can be 3D printed or otherwise manufactured into a custom shape configured to propagate focused ultrasound radiation to one or more focal regions within the target site 30.

[0049] The ultrasound emitter 200 may be configured to deliver acoustic energy at a frequency between 0.5 MHz and 10 MHz to cause heating at the target site 30 .

[0050] In some embodiments, the ultrasound emitter 200 may comprise one or more multi-transducer or phased arrays, planar devices, or bowl-shaped sources. Additionally, the ultrasound emitter 200 may be configured to emit ultrasound radiation interstitially. That is, the ultrasound emitter 200 may comprise one or more catheter-mounted transducers or other emitting members configured to be inserted into the body 35 at or near the target site 30 to emit ultrasound radiation toward one or more locations at the target site 30 and provide the required heating.

[0051] It should be noted that while Figure 2 shows a particular configuration of ultrasound emitters, any suitable ultrasound emitter may be used, for example, any high intensity focused ultrasound (HIFU) device, such as MRI-guided focused ultrasound.

[0052] The ultrasound emitter 200 is configured to emit ultrasound radiation 205 at the target site 30, causing heating at the target site. In particular, the ultrasound emitter 200 is configured to heat the target site to a predetermined temperature, preferably 42°C or less, and maintain the temperature at the predetermined temperature while the electromagnetic field is applied. It will be appreciated that the ultrasound emitter 200 can be configured to heat the target site 30 by providing ultrasound radiation having a predetermined frequency and amplitude that causes the required heating at the target site 30. The ultrasound radiation can be continuous wave or pulsed wave. For a given frequency, amplitude, and type of ultrasound radiation, the amount of heating at the target site can be determined by routine experimentation.

[0053] 2, it will be appreciated that the power of the ultrasound source 202 and the duration of application of the ultrasound radiation may be selected to achieve a predetermined amount of heating at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, with heat being applied by the device to achieve and maintain the target temperature at the target site 30.

[0054] FIG. 3 shows a schematic diagram of an electromagnetic emitter 10 and heat source 300 according to one or more embodiments. As in FIG. 2, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on a surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward a target site 30, resulting in a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0055] The heat source 300 includes one or more antennas or applicators 304 configured to generate an electromagnetic field 305 to directly heat the target site 30. The heat source 300 includes one or more electromagnetic wave sources 302 for driving the one or more antennas or applicators 304 and generating the heating electromagnetic field 305. The electromagnetic field 305 has a field strength and frequency that causes heating of the target site. The frequency of the electromagnetic field 305 is sufficiently different (e.g., by at least one order of magnitude) so that the electromagnetic fields 305 and 15 interact independently with the target site 30 (i.e., so that electromagnetic interference between the fields can be neglected). The frequency of the electromagnetic field 305 can be, for example, greater than 1 MHz to cause molecular dipole rotation, polarization and / or vibration, or dielectric heating of the target site 30 via Ohm's law. The number and configuration of the antennas or applicators 304, including their positions, relative amplitudes, and phases, can be selected to generate constructive and / or destructive interference and cause heating only over a specific volume that includes the target site 30. For example, the antenna or applicator 304 may include a single antenna, a pair of antennas, and a 2D or 3D array or phased array of antennas.

[0056] In some embodiments, the electromagnetic wave source 302 is a radio frequency (RF) source operating at a frequency between 8 MHz and 30 MHz (e.g., 8 MHz, 13.56 MHz, or 27.12 MHz) and configured to induce capacitive heating. The one or more antennas or applicators 304 include a pair of metal applicators, with the target site 30 positioned between them. Optionally, the applicators are connected to a water bolus bag or other medium for transmitting the field to the body 35. When an RF field is applied to the applicators, power is transferred to the target site 30, inducing heating. This technique can be used for both superficial and deep tumors by selecting applicators with different configurations to focus the resulting electric field at the target site 30. Alternatively, the applicators may be mounted coplanar, or one or more applicators may be configured to be positioned within the body 35 within an insulated catheter. A single applicator connected to an external ground plane may alternatively be used. In all of these configurations, the RF field generated at the target site causes direct heating.

[0057] In some embodiments, the electromagnetic source 302 is an RF source configured to operate at a frequency between 60 MHz and 150 MHz. The one or more antennas or applicators 304 include one or more antennas positioned external to the body. Electromagnetic fields generated in this frequency range penetrate deep into the body and are therefore suitable for heating deep target sites 30. Again, the one or more antennas may include a pair of antennas with the target site 30 positioned between them. The antennas may be coupled to a water bolus bag or other medium for delivering the electromagnetic field to the body 35.

[0058] In some embodiments, the electromagnetic wave source 302 is a microwave (MW) source configured to operate at a frequency between 400 MHz and 2.5 GHz (e.g., 433 MHz, 915 MHz, or 2.45 GHz). The one or more antennas may include a pair of antennas or one or more antenna arrays between which the target site 30 is positioned. The antennas may again be coupled to a water bolus bag or other medium for transmitting the electromagnetic field to the body 35.

[0059] It will be appreciated that for the embodiment of Figure 3, the power and application time of the electromagnetic source 302 may be selected to achieve a predetermined amount of heating at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, and heat is applied to achieve and maintain the target temperature at the target site 30.

[0060] FIG. 4 shows a schematic diagram of an electromagnetic emitter 10 and heat source 400 according to one or more embodiments. As in FIGS. 2 and 3, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on a surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward a target site 30, resulting in a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0061] The heat source 400 comprises an electromagnetic wave source 402 and one or more electromagnetic emitters 404 configured to penetrate the body 35 and position a distal portion of the emitter within the target site 30. The one or more emitters 404 are electrically connected to the electromagnetic wave source 402 such that an electrical current is applied to the one or more emitters 404. The one or more emitters 404 may comprise one or more monopole, dipole, slot or helical coil microwave antennas, resistively coupled radio frequency, local current field electrodes, or capacitively coupled radio frequency catheter-based electrodes. Capacitively coupled electrodes may be configured to be housed in a low-loss catheter, such as a nylon or Teflon catheter.

[0062] In some embodiments, the electromagnetic source 402 is configured to supply one or more emitters 404 with alternating current in the frequency range of 350 kHz to 30 MHz, which induces currents in the region of tissue near the needle(s), resulting in tissue heating. In other embodiments, the electromagnetic source 402 is configured to supply one or more emitters 404 with alternating current in the frequency range of 900 MHz to 2.5 GHz (e.g., 915 MHz or 2.45 GHz), which causes dielectric heating of the tissue surrounding the needle(s).

[0063] In another embodiment, the emitter 404 comprises a plurality of electrodes configured to be implanted around the target region 30, and the electromagnetic source 402 is configured to deliver a series of very short (e.g., about 100 μs) direct current electrical pulses between the electrodes. The voltage of the pulses and the electrode configuration are configured to produce high electric field strengths (e.g., about 100 V / cm to 3000 V / cm). Such pulses have been observed to produce heating at the target region 30.

[0064] It will be further appreciated that for the embodiment of Figure 4, the power and application time of the electromagnetic source 402 may be selected to achieve a predetermined amount of heating at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, and heat is applied to achieve and maintain the target temperature at the target site 30.

[0065] FIG. 5 shows a schematic diagram of an electromagnetic emitter 10 and heat source 500 according to one or more embodiments. As with FIGS. 2-4, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on a surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward the target site 30, resulting in a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0066] The heat source 500 comprises one or more infrared light sources 504 configured to provide infrared radiation 505 to the target site 30 and a power supply 502 for driving the one or more infrared lamps. The infrared light sources 504 may emit infrared light of any frequency, particularly frequencies above 300 GHz. Because the penetration depth of infrared radiation is typically 1 cm or less, this device may be suitable for target sites 30 located on the surface of the body 35.

[0067] Additionally, the power output of the power source 502 and the duration of application of the radiation can be selected to achieve a predetermined amount of heating at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, and heat is applied to achieve and maintain the target temperature at the target site 30.

[0068] FIG. 6 shows a schematic diagram of an electromagnetic emitter 10 and heat source 600 according to one or more embodiments. As with FIGS. 2-5, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on a surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward a target site 30, resulting in a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0069] The heat source includes a laser source 604 and a power supply 602 configured to drive the laser source. The laser source 604 may be configured to emit laser radiation at the target site 30, causing heating at the target site. The laser radiation may be optically guided directly to the target site 30 by an optical fiber to cause localized ablation of the target site 30. The laser source 604 may be configured to emit laser radiation having a wavelength between 900 nm and 1100 nm at any intensity suitable for causing the required heating at the target site 30. The laser source 604 may be configured to operate at a power between 0.5 W and 15 W (e.g., 980 nm at 15 W or 1064 nm at 12 W). The laser source 604 may be moved rotationally and linearly to target multiple regions of one or more target sites 30.

[0070] Additionally, the power output of the power source 602 and the duration of application of the laser radiation can be selected to achieve a predetermined amount of heating at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, and heat is applied to achieve and maintain the target temperature at the target site 30. For example, the device may include an MRI device for performing magnetic resonance thermometry to monitor the temperature of the target site 30 during the heating process.

[0071] FIG. 7 shows a schematic diagram of an electromagnetic emitter 10 and a heat source 700 for heating a target site 30 according to one or more embodiments. As with FIGS. 2-6, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on the surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward the target site 30, forming a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0072] The heat source 700 includes a heater 702, a pump 704, a fluid outlet 706, and a fluid inlet 708. The fluid outlet 706 is configured to fluidly connect to a portion of the body 35 upstream of the target site 30, and the fluid inlet 708 is configured to fluidly connect to a portion of the body 35 downstream of the target site 30. The connections form a fluid loop that runs from the target site 30 to the pump 704 and back to the target site 30. The fluid loop may be configured to form in any fluid system of the body 35 (e.g., blood vessels, kidneys, etc.). The heater 702 is configured to heat the fluid to a desired temperature as it passes through the heat source 700, and the fluid is subsequently delivered to the target site 30 via the fluid outlet 706. The fluid loop therefore provides a continuous source of heated fluid to the target site 30. It should be noted that the fluid may be the patient's blood, or additionally, the heat source 700 may include a reservoir of fluid (not shown) configured to be heated and added to the fluid loop. For example, the fluid may contain a chemotherapy composition, an anti-cancer drug, or the like, or may contain a biocompatible solution such as saline.

[0073] Heater 702 may be any suitable heater, such as a resistive heater or an electromagnetic heater configured to heat the fluid by emitting microwave radiation, for example. Heater 702 may be located external to body 35 or may be configured to be implanted within body 35.

[0074] The amount of heat supplied by the heat source 700 and the time of application of the heat source 700 can be selected to achieve a predetermined amount of heat at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, and heat is applied to achieve and maintain the target temperature at the target site 30.

[0075] FIG. 8 shows a schematic diagram of an electromagnetic emitter 10 and a heat source 800 for heating a target site 30 according to one or more embodiments. As with FIGS. 2-7, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on the surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward the target site 30, resulting in a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0076] Heat source 800 comprises heater 802, reservoir 805, pump 804, and fluid outlet 806. Fluid outlet 806 is configured to be fluidly connected to target site 30. In use, pump 804 is configured to pump fluid in reservoir 805 to target site 30 via fluid outlet 806. Heater 802 is configured to heat fluid in reservoir 805 to a desired temperature before the fluid is pumped to target site 30. The fluid may include a chemotherapeutic composition, an anti-cancer drug, etc., or may include a biocompatible solution such as saline.

[0077] The amount of heat supplied by heat source 800, the amount of fluid supplied from reservoir 805, and the duration of application of heat source 800 can be selected to achieve a predetermined amount of heating at target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of target site 30, and heat is applied to achieve and maintain the target temperature at target site 30.

[0078] FIG. 9 shows a schematic diagram of an electromagnetic emitter 10 and a heat source 900 for heating a target site 30 according to one or more embodiments. As with FIGS. 2-8, the electromagnetic emitter 10 may include one or more electromagnetic wave sources 12 (e.g., one or more current or voltage sources) electrically connected to one or more applicators 14. The applicators 14 may be configured to be positioned on a surface of a patient's body 35. The electromagnetic wave sources 12 may be configured to provide an alternating current electromagnetic field to the applicators 14, which generates a magnetic flux density toward the target site 30, resulting in a non-ionizing alternating current electromagnetic field 15 at the target site 30. It will be appreciated that any number of applicators 14 may be used depending on the type of target site, and the strength of the resulting magnetic flux density at the target site 30 may be readily calculated from the superposition of the electromagnetic fields emitted by each applicator 14.

[0079] The heat source 900 includes a heat emitter 904 configured to provide conductive heating to the target site 30. The heat source may include a power source 902 (such as an electrical power source) for driving the heat emitter 904, or the heat emitter 904 may be preheated or chemically self-heated (e.g., via an exothermic chemical reaction). The heat emitter 904 may be provided on the surface of the body 35 or may be implanted within the body and configured to provide heat to the target site 30 at a location proximal to the target site. For example, the heat emitter 904 may be an implantable resistive heater configured to be powered by the electrical power source 902. The heat emitter 904 may be configured to heat a localized region of the body 35 or may be configured to heat the entire body 35.

[0080] The amount of heat supplied by the heat source 900 and the time of application of the heat source 900 can be selected to achieve a predetermined amount of heat at the target site 30. This may be determined by prior empirical measurement, or the device may additionally include a thermometer element for measuring the temperature of the target site 30, and heat is applied to achieve and maintain the target temperature at the target site 30.

[0081] FIG. 10 illustrates an overview of an apparatus 1 for treating a cancerous target site according to one or more embodiments. As in FIG. 1, the apparatus includes an electromagnetic emitter 10 and a heat source 20. The electromagnetic emitter 10 is configured to provide a magnetic flux density at the target site. The heat source 20 is configured to provide direct heating at the target site by any of the mechanisms disclosed herein above, inducing hyperthermia at the target site. In one configuration, the apparatus 1 is configured to simultaneously provide a non-ionizing alternating electromagnetic field and direct heating at the target site. The electromagnetic emitter 10 and heat source 20 can have any suitable configuration, including those shown in FIGS. 2-9.

[0082] The apparatus 1 shown in FIG. 10 further comprises a controller 40 for controlling the emitter 10 and the heat source 20. The controller comprises a first control interface 41 for controlling the operation of the electromagnetic emitter 10 and a second control interface 42 for controlling the operation of the heat source 20. The controller 40 is configured to provide control signals to the emitter 10 via the control interface 41 and to the heat source 20 via the control interface 42. The emitter 10 and the heat source 20 may receive the control signals via any suitable form of communication, either wired or wireless, such as optical, fiber optic, Ethernet, or any suitable wireless communication. Furthermore, the controller 40 may be configured to drive one or more of the emitter 10 and the heat source 20, or one or more of the emitter 10 and the heat source 20 may be driven independently of the controller 40.

[0083] The controller 40 further comprises one or more of a user interface 43, a memory 44, and a processor 45. The user interface 43 allows a user to manually control the operation of the emitter 10 and heat source 20, for example, by controlling operating parameters of the emitter 10 and heat source 20 and turning their operation on or off. The user interface 43 may allow a user to select a sequence of operation of the emitter 10 and heat source 20 over a period of time. The memory 44 may include instructions that, when executed by the processor 45, cause the emitter 10 and heat source 20 to operate according to any suitable sequence, including the sequences of operation disclosed herein.

[0084] Memory 44 may include one or more volatile or non-volatile memory devices, such as DRAM, SRAM, flash memory, read-only memory, ferroelectric RAM, hard disk drives, floppy disks, magnetic tape, optical disks, etc. Similarly, processor 45 may include one or more processing devices, such as a microprocessor, GPU, CPU, multi-core processor, etc. Furthermore, controller 40 may be implemented in software, hardware, or any combination to perform the sequences of operations disclosed herein.

[0085] Figure 11 illustrates an overview of a method for treating a cancerous target site using a device according to the present disclosure. The device can have any of the configurations disclosed herein. Prior to performing the method of Figure 11, the emitter 10 and heat source 20 can be appropriately positioned on the patient. For example, the applicator of the electromagnetic emitter 10 can be placed at a predetermined location on the patient's body, and the heat source 20 (e.g., transducer 24) can also be appropriately positioned.

[0086] In step S1, an electromagnetic emitter 10 is used to provide a non-ionizing alternating electromagnetic field, generating a magnetic flux density at the target site.

[0087] In step S2, heat (more specifically, direct heat) is supplied to the target site using heat source 20. This can be supplied simultaneously with, or within a predetermined time period before or after, the application of the electromagnetic field to target site 30. In some embodiments, an alternating electromagnetic field alone may be applied without heating for a first period of time to create a magnetic flux density at the target site, and then both may be applied to the target site simultaneously.

[0088] Optionally, in step S3, an anti-tumorigenic composition is delivered to the target site. The anti-tumorigenic composition can be administered by any suitable means, such as orally or intravenously. It should be noted that the anti-tumorigenic composition can be delivered before or simultaneously with steps S1, S2, and S4, or a predetermined time after step S1, S2, or S4.

[0089] The term "anti-tumorigenic composition," as used herein, refers to a composition containing an agent that at least partially inhibits the development or progression of cancer, including inhibiting all or some of the symptoms associated with cancer. The term "cancer," as used herein, refers to a disease characterized by uncontrolled cell division (or increased survival or resistance to apoptosis) and the ability of such cells to invade other adjacent tissues (invasion), spread through lymphatic and blood vessels to other areas of the body where the cells are not normally found (metastasize), and, after circulating in the bloodstream, invade normal tissues in other parts of the body. Tumors are classified as either benign or malignant depending on whether they can spread by invasion and metastasis. Benign tumors are tumors that cannot spread by invasion or metastasis, i.e., grow only locally, while malignant tumors are tumors that can spread by invasion and metastasis. The anti-tumorigenic composition can include one or more anti-tumorigenic compositions, including one or more of those disclosed with respect to Figures 12-27.

[0090] As used herein, the term cancer preferably refers to solid and / or invasive tumors.

[0091] As used herein, a "solid tumor" is understood to be an abnormal mass of tissue that usually does not contain cysts or liquid areas. Various types of solid tumors are named for the type of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors.

[0092] As used herein, an "invasive tumor" is understood to be a tumor with an abnormal tumor structure that simultaneously exhibits obvious growing nodules and infiltrative growth.

[0093] Preferably, the present invention is directed to cancers including, but not limited to, the following types: breast cancer; biliary tract cancer; bladder cancer; brain cancer, including glioblastoma, particularly glioblastoma multiforme and medulloblastoma; cervical cancer; head and neck cancer; choriocarcinoma; colon cancer, colorectal cancer; endometrial cancer; esophageal cancer; gastric cancer; intraepithelial neoplasia, including Bowen's disease and Paget's disease; liver cancer, hepatocellular carcinoma; lung cancer, pleural mesothelioma; oral cancer, including squamous cell carcinoma; parotid gland cancer; ovarian cancer, including those arising from epithelial, stromal, germinal and mesenchymal cells; pancreatic cancer; prostate cancer; kidney cancer rectal cancer; sarcoma, including leiomyosarcoma, rhabdomyosarcoma, liposarcoma, fibrosarcoma, and osteosarcoma; skin cancer, including melanoma, Merkel cell carcinoma, Kaposi's sarcoma, basal cell carcinoma, and squamous cell carcinoma; cervical cancer, endometrial cancer; testicular cancer, including germ cell tumors such as seminoma, non-seminoma (teratoma, choriocarcinoma), stromal tumors, and germ cell tumors; thyroid cancer, including thyroid adenocarcinoma and medullary carcinoma; and renal cancer, including adenocarcinoma and Wilms' tumor.

[0094] In certain embodiments, the cancer is melanoma. As used herein, the term "melanoma" refers to malignant skin tumors of melanocytes, including, but not limited to, melanoma, metastatic melanoma, melanoma derived from either melanocytes or melanocyte-associated nevus cells, malignant melanoma, melanoepithelioma, melanosarcoma, intraepithelial melanoma, superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, acral lentigo melanoma, invasive melanoma, and familial atypical nevus melanoma (FAM-M) syndrome. Furthermore, the term "melanoma" refers not only to primary melanoma, but also to "melanoma metastasis," which, as used herein, refers to the spread of melanoma cells to regional lymph nodes and / or distant organs. This event occurs frequently, given that melanoma contains multiple cell populations characterized by diverse growth rates, karyotypes, cell surface properties, antigenicity, immunogenicity, invasion, metastasis, and sensitivity to cytotoxic or biological agents. Melanoma frequently metastasizes to the brain, lungs, lymph nodes, and skin. Other cancers will be known to those skilled in the art.

[0095] Figure 12 shows experimental data demonstrating the in vitro effects of TT domain ("TTF"), hyperthermia ("HT"), pterostilbene ("PT"), and their combination on U87MG cells. The TT domain was applied at 300 kHz for 240 minutes (from 0 to 240 minutes) with an average magnetic flux density of 8 μT. The hyperthermia was applied at 42°C for 10 minutes (from 120 to 130 minutes). 20 μM pterostilbene was applied for 120 minutes (from 120 to 240 minutes). Data represent the average number of viable cells for five experiments, with * versus control, + versus TTF, and # versus TTF + HT, P<0.01 using Student's t-test. The magnetic field was applied using a solenoid with a magnetic flux density of approximately 100 μT at the central axis of the solenoid. The solenoid was positioned a short distance from the cell culture so that the average electric field strength on the surface of the culture flask to which the cells were attached was 8 μT.

[0096] As already shown above and as can be seen in Figure 12, it should be noted that hyperthermia was found to enhance the anti-cancer effects of non-ionizing alternating electromagnetic fields (TT-region). Notably, this combination allows for much lower hyperthermia temperatures (42°C or lower) while reducing cell viability. This also means that a larger volume of tissue can be heated and targeted by hyperthermia. Thus, the device disclosed herein provides an effective method for treating cancerous sites by applying TT-region and direct heating (i.e., without mediators) of the cancerous site. Furthermore, as shown in Figure 12, the combination of TT-region, hyperthermia, and pterostilbene effectively eliminates all cells in vitro. This combination therapy is not expected to have any substantial side effects in vivo, as the dose of pterostilbene used is well tolerated in vivo.

[0097] Figure 13 shows in vitro experimental data on the effects of exposure to various oscillating magnetic fields on cell viability of U87MG (ATCC) cells. Various cell cultures were exposed to one of the following magnetic fields: 24 μT at a frequency of 100 kHz, 12 μT at a frequency of 200 kHz, 8 μT at a frequency of 300 kHz, or 6 μT at a frequency of 400 kHz. Cell viability for each frequency was measured after 1, 2, 3, 4, and 5 hours. Five independent experiments were performed for each frequency and time point. Data for each frequency represent the mean cell viability and standard deviation over time (from left to right, 1 hour to 5 hours) for five corresponding experiments. Comparisons between different groups were performed using a two-way analysis of variance (ANOVA). It can be seen that cell viability decreased at all frequencies, with the effect further increasing at frequencies below 300 kHz. The letters "a" through "f" were assigned to the data based on the statistical tests applied to the data. Data marked with the same letter are considered statistically similar, and data assigned different letters are considered significantly different at P less than 0.01.

[0098] Figure 14 shows in vitro experimental data on the effect of heat exposure on cell viability of U87MG cells. Various cell cultures were heated to temperatures of 37°C, 38°C, 39°C, 40°C, 41°C, or 42°C. Five independent experiments were performed for each temperature and time point. Data for each temperature represent the mean cell viability and standard deviation of five corresponding experiments after 5 and 10 minutes of exposure to that temperature. Hyperthermia, equivalent to very high heat (41°C or higher), significantly reduced U87MG cell viability. Data marked with * indicate a P value of less than 0.01 (Student's t-test) compared to the 37°C data at the corresponding time, and data marked with + indicate a P value of less than 0.01 for the 10-minute data compared to the 5-minute data at the same temperature.

[0099] Figures 15B-15F show microscopic images of in vitro U87MG cell cultures after exposure to different external treatments. Figure 15A shows a microscopic image of a control in vitro U87MG cell culture that was not exposed to any external treatment and maintained at a physiological internal temperature of 37°C. Figure 15B shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 8 μT at 300 kHz from 0 to 240 minutes. Figure 15C shows the cell culture after exposure to heating to 42°C for 10 minutes from 120 to 130 minutes. Figure 15D shows the cell culture after exposure to 20 μM pterostilbene from 120 to 240 minutes. Figure 15E shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 8 μT at 300 kHz from 0 to 240 minutes and exposure to 20 μM pterostilbene from 120 to 240 minutes. Figure 15F shows cell cultures after exposure to an electromagnetic field with a magnetic flux density of approximately 8 μT at 300 kHz from 0 to 240 minutes, exposure to heating to 42°C for 10 minutes from 120 to 130 minutes, and exposure to 20 μM pterostilbene (PT) from 120 to 240 minutes. Note that the combination of TTF+HT+PT completely eliminates all U87MG proliferating cells. This therapy shows the same efficacy in other glioblastoma lines, such as C6 and GL261.

[0100] FIG. 16 shows in vitro experimental data on the effects on cell viability of U87MG cells upon exposure to an electromagnetic field having a magnetic flux density of approximately 8 μT at 300 kHz for approximately 240 minutes (data labeled "TTF"), a temperature of 42° C. for approximately 10 minutes (data labeled "HT"), 50 μM temozolomide (data labeled "TMZ"), and a combination thereof. Data represent the mean of five independent experiments. Data labeled with * indicate a P value of less than 0.01 (Student's t-test) compared to the control. Data labeled with + indicate a P value of less than 0.01 (Student's t-test) compared to the TTF-only data. Data labeled with # indicate a P value of less than 0.01 (Student's t-test) compared to the TTF+HT data.

[0101] FIG. 17 shows the results of applying an electromagnetic field having a magnetic flux density of about 8 μT at 300 kHz for about 240 minutes (data labeled "TTF"), a temperature of 42° C. for about 10 minutes from 120 to 130 minutes (data labeled "HT"), 20 μM resveratrol (data labeled "R") from 210 to 240 minutes, 20 μM resveratrol triphosphate (data labeled "R-triP") from 210 to 240 minutes, 20 μM 4'-butyrate-3,5-dihydroxystilbene (data labeled "B-diOH-s") from 210 to 240 minutes, 20 μM 3-glucoside-5,4'-dihydroxystilbene (data labeled "G-diOH-s") from 210 to 240 minutes, and 20 μM Figure 1 shows in vitro experimental data on the effect of exposure to 3-amido-5,4'-dihydroxystilbene (data labeled "A-diOH-s") and their combinations on cell viability of U87MG cells. Data are the mean + standard deviation for four independent experiments. Data labeled with * indicate a P value of less than 0.01 (Student's t-test) compared to the control. Data labeled with + indicate a P value of less than 0.01 (Student's t-test) compared to the TTF-only data. Data labeled with # indicate a P value of less than 0.01 (Student's t-test) compared to the TTF+HT data. It is observed that a significant decrease in cell viability occurs with TTF+HT+G-diOH-s, but resveratrol and its derivatives do not eliminate all U87MG cell proliferation in vitro.

[0102] The efficacy of heat therapy in combination with tumor treatment area and PT exposure has also been observed in other cell lines in vitro, such as A2058 (melanoma), AsPC-1 (pancreatic cancer), A549 (lung cancer), MCF-7 (breast adenocarcinoma), HT-29 (colon cancer), PC-3 (prostate cancer), SK-OV-3 (ovarian cancer), and HepG2 (hepatocellular carcinoma).

[0103] Figure 18 shows in vitro experimental data on the effects of exposure to various oscillating magnetic fields on cell viability of AsPC1 (pancreatic adenocarcinoma, ATCC) cells. Various cell cultures were exposed to one of the following magnetic fields: 2 μT at 100 kHz, 1.5 μT at 200 kHz, 0.7 μT at 300 kHz, or 0.5 μT at 400 kHz. Cell viability for each frequency was measured for each cell culture after 1, 2, 3, 4, and 5 hours. Five independent experiments were performed for each frequency and time point. Data for each frequency represent the mean cell viability and standard deviation over time (1 hour to 5 hours, from left to right) for five corresponding experiments. Comparisons between different groups were performed using two-way analysis of variance (ANOVA). Cell viability is seen to decrease at frequencies below 200 kHz. The letters "a" and "b" were assigned to the data based on the statistical tests applied to the data. Data marked with the same letter are considered statistically similar, and data assigned different letters are considered significantly different at P less than 0.01.

[0104] Figure 19 shows in vitro experimental data on the effect of heat exposure on cell viability of AsPC1 cells. Various cell cultures were heated to temperatures of 37°C, 42°C, 47°C, or 52°C. Five independent experiments were performed for each temperature and time point. Data for each temperature represent the mean cell viability and standard deviation of five corresponding experiments after 5, 10, and 20 minutes of exposure to that temperature. Heating to 47°C or higher significantly reduced AsPC1 cell viability. Data marked with * indicate a P value of less than 0.01 (Student's t-test) compared to the 37°C data at the corresponding time, and data marked with + indicate a P value of less than 0.01 for the 10- and 20-minute data compared to the 5-minute data at the same temperature.

[0105] FIG. 20 shows experimental data demonstrating the in vitro effects of TT domain ("TTF"), hyperthermia ("HT"), pterostilbene ("PT"), and their combination on AsPC1 cells. For the TTF data, the applied TT domain was 200 kHz at a field strength of 1.5 μT for 240 minutes (from 0 to 240 minutes). For the HT data, the applied hyperthermia was 52°C for 20 minutes (from 120 to 140 minutes). For the PT data, 20 μM pterostilbene was applied from 0 to 240 minutes. Data represent the average number of viable cells for five experiments, with * indicating vs. control, + indicating vs. TTF, and # indicating vs. TTF + HT, P<0.01 using Student's t-test.

[0106] Figures 21B-21H show microscopic images of in vitro AsPC1 cell cultures after exposure to different external treatments. Figure 21A shows a microscopic image of a control vitro AsPC1 cell culture that was not exposed to any external treatment and was maintained at the optimal temperature for AsPC1. Figure 21B shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes. Figure 21C shows the cell culture after exposure to heating to 52°C for 20 minutes from 120 to 140 minutes. Figure 21D shows the cell culture after exposure to 20 μM pterostilbene from 0 to 240 minutes. Figure 21E shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes and heating to 52°C for 20 minutes from 120 to 140 minutes. Figure 21F shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes, and after exposure to 20 μM pterostilbene (PT) from 0 to 240 minutes. Figure 21G shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes, exposure to heating to 52°C for 20 minutes from 120 to 140 minutes, and after exposure to 20 μM pterostilbene (PT) from 0 to 240 minutes. Figure 21H shows the cell culture of Figure 21G after 24 hours, during which the cultured cells were maintained at 37°C without any treatment. Proliferating cells do not appear to recover after 24 hours of combined treatment in Figure 21G.

[0107] FIG. 22 shows experimental data demonstrating the in vitro effects of TTTF ("TTF"), hyperthermia ("HT"), gemcitabine ("GEM"), pterostilbene ("PT"), and their combinations on AsPC1 cells. For the TTF data, the applied TTTF was at 200 kHz for 240 minutes (from 0 to 240 minutes) at a field strength of 1.5 μT. For the HT data, the applied hyperthermia was at 52° C. for 20 minutes (from 120 to 140 minutes). For the GEM data, 25 μM gemcitabine was applied from 0 to 240 minutes. For the PT data, 20 μM pterostilbene was applied from 0 to 240 minutes. Data show the average number of viable cells for five experiments per experimental condition, * vs. control, + vs. TTF, # vs. TTF+HT using Student's t-test, P<0.01. All four treatments applied in combination eliminated AsPC1 cells.

[0108] Figure 23 shows in vitro experimental data on the effect of exposure to different oscillating magnetic fields on cell viability of A2058 (melanoma, ATCC) cells. Various cell cultures were exposed to one of the following magnetic fields: 2 μT at a frequency of 100 kHz, 1.5 μT at a frequency of 200 kHz, 0.7 μT at a frequency of 300 kHz, or 0.5 μT at a frequency of 400 kHz. Cell viability for each frequency was measured for each cell culture after 1, 2, 3, 4, and 5 hours. Five independent experiments were performed for each frequency and time point. Data for each frequency represent the mean cell viability and standard deviation over time (from left to right, 1 hour to 5 hours) for five corresponding experiments. Comparisons between different groups were performed using a two-way analysis of variance (ANOVA). It can be seen that cell viability decreases at all frequencies below 300 kHz. The letters "a" and "b" were assigned to the data based on the statistical tests applied to the data. Data marked with the same letter are considered statistically similar, and data assigned different letters are considered significantly different at P less than 0.01.

[0109] Figure 24 shows in vitro experimental data on the effect of heat exposure on cell viability of A2058 cells. Various cell cultures were heated to temperatures of 37°C, 42°C, 47°C, or 52°C. Five independent experiments were performed for each temperature and time point. Data for each temperature represent the mean cell viability and standard deviation of five corresponding experiments after 5, 10, and 20 minutes of exposure to that temperature. Heating to 52°C significantly reduced A2058 cell viability. Data marked with * indicate a P value of less than 0.01 (Student's t-test) compared to the 37°C data at the corresponding time, and data marked with + indicate a P value of less than 0.01 for the 10- and 20-minute data compared to the 5-minute data at the same temperature.

[0110] Figure 25 shows experimental data demonstrating the in vitro effects of TT domain ("TTF"), hyperthermia ("HT"), pterostilbene ("PT"), and their combination on A2058 cells. For the TTF data, the applied TT domain was 200 kHz at a field strength of 1.5 μT for 240 minutes (from 0 to 240 minutes). For the HT data, the applied hyperthermia was 52°C for 20 minutes (from 120 to 140 minutes). For the PT data, 20 μM pterostilbene was applied from 0 to 240 minutes. Data represent the average number of viable cells for five experiments per experimental condition, with * indicating a difference vs. control, + indicating a difference vs. TTF, and # indicating a difference vs. TTF + HT, P<0.01 using Student's t-test. Application of all four treatments significantly reduced cell viability compared to TTF + HT.

[0111] Comparing the data in Figures 12, 20, and 25, the data show that using higher magnetic flux densities (above about 8 μT) allows tissue to be heated to lower temperatures while maintaining or even improving treatment effectiveness. The combination of such higher magnetic flux densities in combination with heating to temperatures above 42°C and administration of pterostilbene may be sufficient to eliminate tumors. This is particularly important for areas of the body where only limited heating can be applied (e.g., the brain). For example, the data in Figure 22 show that even with lower magnetic fields, tumor treatment areas and heating may be used in combination with pterostilbene and one or more other anti-cancer drugs to reduce or even eliminate cancer cells.

[0112] Figures 26B-26H show microscopic images of in vitro A2058 cell cultures after exposure to different external treatments. Figure 26A shows microscopic images of a control in vitro AsPC1 cell culture that was not exposed to any external treatments and maintained at a physiological internal temperature of 37°C. Figure 26B shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes. Figure 26C shows the cell culture after exposure to heating to 52°C for 20 minutes from 120 to 140 minutes. Figure 26D shows the cell culture after exposure to 20 μM pterostilbene from 0 to 240 minutes. Figure 26E shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes and exposure to 20 μM pterostilbene from 0 to 240 minutes. Figure 26F shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes, and heating to 52°C for 20 minutes from 120 to 140 minutes. Figure 26G shows the cell culture after exposure to an electromagnetic field having a magnetic flux density of approximately 1.5 μT at 200 kHz from 0 to 240 minutes, heating to 52°C for 20 minutes from 120 to 140 minutes, and exposure to 20 μM pterostilbene (PT) from 0 to 240 minutes. Figure 26H shows the cell culture of Figure 26G after 24 hours, during which the cultured cells were maintained at 37°C without any treatment. Proliferating cells do not appear to recover after 24 hours of combined treatment in Figure 26G.

[0113] Figure 27 shows experimental data demonstrating the in vitro effects of TTTF ("TTF"), hyperthermia ("HT"), paclitaxel ("PAC"), and their combinations on A2058 cells. For the TTF data, the applied TTTF was at 200 kHz for 240 minutes (from 0 to 240 minutes) at a field strength of 1.5 μT. For the HT data, the applied hyperthermia was at 52°C for 20 minutes (from 120 to 140 minutes). For the PAC data, 10 μM paclitaxel was applied from 0 to 240 minutes. Data represent the average number of viable cells for five experiments per experimental condition, with * indicating vs. control, + indicating vs. TTF, and # indicating vs. TTF + HT, P<0.01 using Student's t-test. Application of all three treatments resulted in cell elimination.

[0114] Table 1 shows data examining the effect of GSH depletion on cell viability in U87MG, AsPC1, and A2058 cells in vitro. For rows containing "TTF," the TT regime was applied at 200 kHz for 240 minutes (from 0 to 240 minutes) at a field strength of 1.5 μT. For rows containing "HT," hyperthermia was applied at 42°C for U87MG and 52°C for AsPC1 and A2058 for 20 minutes (from 120 to 140 minutes). For rows containing "BSO," 1 mM buthionine sulfoximine, a specific inhibitor of GSH synthesis, was added to the cell culture medium at the time the cultures were seeded. When HT and TTF were applied in combination with BSO, HT and TTF were applied 24 hours after seeding. Data represent the mean and standard deviation for five independent experiments. Data marked with * have a P value of less than 0.01 (Student's t-test) compared to the control data. Data marked with + have a P value (Student's t-test) less than 0.01 compared to the TTF+HT data. The data show that GSH depletion enhances the anti-cancer effects of electromagnetic radiation and heat.

[0115] [Table 1]

[0116] Therefore, the use of the device disclosed herein in combination with the application of pterostilbene provides a highly effective method for treating cancerous sites, even completely eliminating cancerous cells. Thus, in one embodiment, the anti-tumorigenic composition comprises (i) pterostilbene, pterostilbene phosphate, or a pharmaceutically acceptable salt thereof. Alternatively, the anti-cancer composition may be provided in a cocrystal, water-soluble prodrug, nanoparticle, nanodot, nanorob, nanospike, nanorod, nanocluster, nanoceramic, liposome, or exosome formulation, or in an implantable device configured to release the anti-cancer composition when implanted in the body. It should be noted that the anti-tumorigenic composition may comprise any antioxidant composition. In this sense, the anti-tumorigenic composition may comprise, apart from pterostilbene, any stilbenoid suitable as an anti-tumorigenic agent, such as resveratrol.

[0117] (i) Pterostilbene and pterostilbene phosphate The term "pterostilbene" or "Pter" or "trans-3,5-dimethoxy-4'-hydroxystilbene" as used herein refers to a compound of the formula: [ka] This refers to the compound.

[0118] The term "pterostilbene phosphate" means a compound of the formula: [ka] This refers to the compound.

[0119] The term "pharmaceutically acceptable salt" refers to any salt of pterostilbene or pterostilbene phosphate that can provide (directly or indirectly) the compounds described herein after administration to a recipient.Preferably, as used herein, the term "pharmaceutically acceptable salt" means that it is approved by a federal or state government regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, more particularly in humans.Salts can be prepared by methods known in the art. Illustrative non-limiting examples of pharmaceutically acceptable salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, hydrogen sulfate, phosphate, superphosphate, isonicotinate, lactate, salicylate, percitrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate. Pharmaceutically acceptable salts of pterostilbene or pterostilbene phosphate are preferably prepared from a polyphenol compound having an acidic functional group and an acceptable inorganic or organic base.Suitable bases include, but are not limited to, hydroxides of alkali metals such as sodium, potassium, and lithium; hydroxides of alkaline earth metals such as calcium and magnesium; hydroxides of other metals such as aluminum and zinc; ammonia and organic amines such as unsubstituted or hydroxy-substituted mono-, di-, or tri-alkylamines, dicyclohexylamine; tributylamine; pyridine; N-methylamine; N-ethylamine; diethylamine; triethylamine; mono-, bis-, or tris-(2-hydroxy-substituted lower alkylamines) such as mono-, bis-, or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine, or tris-(hydroxymethyl)methylamine; N,N-di-lower alkyl-N-(hydroxy lower alkyl)amines such as N,N-dimethyl-N-(2-hydroxyethyl)amine or tri-(2-hydroxyethyl)amine; N-methyl-O-glucamine; and amino acids such as arginine and lysine. The term "pharmaceutically acceptable salt" also includes hydrates of polyphenolic compounds. In certain embodiments, the pharmaceutically acceptable salt is a disodium salt.

[0120] Further illustrative, non-limiting examples of cancer chemotherapeutic agents that can be used in accordance with the present invention include alkylating agents such as nitrogen mustards / oxazaphosphorines (e.g., cyclophosphamide, ifosfamide), nitrosoureas (e.g., carmustine), triazenes (e.g., temozolamide), and alkyl sulfonates (e.g., busulfan); antimetabolites (e.g., 5-fluorouracil, capecitabine, 6-mercaptopurine, methotrexate, gemcitabine, cytarabine, fludarabine, or pemetrexed); doxorubicin and daunol. Anthracycline antibiotics such as bicine, taxanes such as Taxol™ and docetaxel, vinca alkaloids such as vincristine and vinblastine, 5-fluorouracil (5-FU), leucovorin, irinotecan, idarubicin, mitomycin C, oxaliplatin, raltitrexed, pemetrexed, tamoxifen, cisplatin, carboplatin, methotrexate, actinomycin D, mitoxantrone, blenoxane, mithramycin, paclitaxel, 2-methoxyestradiol, prinomastat, batimastat, BAY 12-9566, carboxyamidotriazole, CC-1088, dextromethorphan acetate, dimethylxanthenone acetate, endostatin, IM-862, marimastat, penicillamine, PTK787 / ZK 222584, RPI.4610, squalamine lactate, SU5416, thalidomide, combretastatin, COL-3, neovastat, BMS-275291, SU6668, anti-VEGF antibody, Medi-522 (Vitaxin II), CAI, interleukin-12, IM862, amiloride, angiostatin, angiostatin K1-3, angiostatin K1-5, captopril, DL-α-difluoromethylornithine, DL-α-difluoromethylornithine HCl, endostatin, fumagillin, herbimycin A, 4-hydroxyphenylretinamide, juglone, laminin, laminin hexapeptide, laminin pentapeptide, lavendustin A, medroxyprogesterone, minocycline, placental ribonuclease inhibitor, suramin, thrombospondin, antibodies targeting pro-angiogenic factors (e.g., bevacizumab, cetuximab, panitumumab, trastuzumab); topoisomerase inhibitors (e.g., thrombospondin, thrombospondin); Examples of suitable chemotherapeutic agents include erythromycinase inhibitors; microtubule inhibitors; small molecular weight tyrosine kinase inhibitors of pro-angiogenic growth factors (e.g., erlotinib, sorafenib, sunitinib, gefitinib); GTPase inhibitors; histone deacetylase inhibitors; AKT kinase or ATPase inhibitors; Wnt signaling inhibitors; inhibitors of E2F transcription factors; mTOR inhibitors (e.g., temsirolimus); α, β, and γ interferon, IL-12, matrix metalloproteinase inhibitors (e.g., COL3, marimastat, batimastat); ZD6474, SU11248, vitaxin; PDGFR inhibitors (e.g., imatinib); NM3 and 2-ME2; and cyclic peptides such as cilengitide. Other suitable chemotherapeutic agents are described in detail in The Merck Index in CD-ROM, 13th Edition. In a preferred embodiment of the invention, the chemotherapeutic agent is selected from the group consisting of docetaxel (Taxotere™), cisplatin, pemetrexed, gemcitabine, and irinotecan.

[0121] In certain embodiments, the cancer chemotherapeutic agent is a taxane, preferably comprising or consisting of paclitaxel. The term "paclitaxel" as used herein has the chemical name (2α,4α,5β,7β,10β,13α)-4,10-bis(acetyloxy)-13-{[(2R,3S)-3(benzoylamino)-2-hydroxy-3-phenylpropanoyl]oxy}-1,7-dihydroxy-9-oxo-5,20-epoxytax-11-en-2-ylbenzoate and the chemical formula: [ka] It refers to a compound having the formula:

[0122] In more particular embodiments, the paclitaxel is protein-bound paclitaxel. The terms "protein-bound paclitaxel" or "nab-paclitaxel" or "nanoparticulate albumin-bound paclitaxel," as used herein, refer to a formulation in which paclitaxel is bound to albumin as a delivery vehicle.

[0123] The cancer chemotherapeutic agent will vary depending on the type of cancer being treated by the combination of the present invention. One skilled in the art can readily determine which cancer chemotherapeutic agent is more suitable for treating a particular type of cancer.

[0124] Optionally, in step S4, a glutathione (GSH)-depleting agent is provided to the target site in addition to the anti-tumorigenic compound. The GSH-depleting agent can be administered by any suitable means, such as orally or intravenously. Note that the GSH-depleting agent can be provided prior to or simultaneously with steps S1, S2, and S3, or a predetermined time after step S1, S2, or S3, preferably a predetermined time after step S3. The GSH-depleting agent provided can be any GSH-depleting agent disclosed herein, including in connection with Figures 12-27 or Table 1.

[0125] The term "glutathione depleting agent," as used herein, refers to a substance that reduces or eliminates glutathione from cells that come into contact with the substance. Those skilled in the art can determine whether a particular molecule is a glutathione depleting agent, for example, by comparing the effect of the particular molecule with that of buthionine sulfoximine (BSO), a specific inhibitor of γ-glutamyl-cysteinyl ligase, using the method described by Terradez P et al., Biochem J 1993, 292 (Pt 2): 477-83, for in vitro and in vivo conditions. In certain embodiments, a particular molecule is a glutathione depleting agent if it has at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, 100% or more of the glutathione-depleting effect of buthionine sulfoximine. Illustrative, non-limiting examples of glutathione depleting agents are as follows:

[0126] a) Bcl-2 antisense oligodeoxynucleotides, i.e., oligodeoxynucleotides complementary to the RNA sequence of the Bcl-2 gene, as described in Ortega, et al., Cancers (Basel) 2011, 3, 1285-1310. Non-limiting examples of Bcl-2 antisense oligodeoxynucleotides are described in U.S. Pat. No. 5,734,033, WO 2003040182, and U.S. Pat. No. 5,831,066. Assays to determine whether a particular compound is a Bcl-2 antisense oligodeoxynucleotide are based on the compound's effect on Bcl-2 mRNA levels or Bcl-2 protein levels, as described, for example, in Mena et al., Clinical Cancer Research 2007, 13 (9): 2658-66.

[0127] b) Inhibitors of multidrug resistance protein 1 (MRP1), such as those described in Ortega et al. (supra). The term "MRP1 inhibitor," as used herein, refers to a compound that inhibits the activity of MRP1. The term inhibitor includes, but is not limited to, MRP1 antagonists, antibodies to MRP1, compounds that interfere with the expression of MRP1, and compounds that reduce MRP1 mRNA or protein levels. Non-limiting examples of MRP1 inhibitors are verapamil and MK-571. An assay for determining whether a particular compound is an MRP1 inhibitor is, for example, the method described in Olson DP et al., Cytometry 2001, 46 (2): 105-13.

[0128] c) Inhibitors of gamma-glutamine transpeptidase or gamma-glutamyltransferase (GGTP or GGT), such as those described in Silber et al., Anal Biochem 1986, 158 (1): 68-71. The term "GGTP inhibitor," as used herein, refers to a compound that inhibits the activity of GGTP, an enzyme that catalyzes the transfer of the gamma-glutamyl moiety of glutathione to an acceptor. The term inhibitor includes, but is not limited to, GGTP antagonists, antibodies against GGTP, compounds that interfere with GGTP expression, and compounds that reduce GGTP mRNA or protein levels. GGTP inhibitors include both selective and non-selective inhibitors (which also affect asparagine synthetase). Non-limiting examples of GGTP inhibitors are acivicin and 2-amino-4-{[3-(carboxymethyl)phenyl](methyl)phosphono}butyric acid (GGsTop™). Assays to determine whether a particular compound is a GGTP inhibitor are, for example, those described in Silver et al., Anal Biochem 1986, 158 (1): 68-71.

[0129] d) Inhibitors of cystine uptake, such as those described in Obrador, et al., Hepatology 2002, 35, 74-81. The term "inhibitors of cystine uptake" refers to sodium-independent X-linked cystine inhibitors. c The term "inhibitor" refers to a compound that inhibits any of the systems that transport extracellular cystine into cells, including the sodium-dependent XAG system and the sodium-dependent XAG system (McBean GJ and Flynn J., Biochem Soc Trans. 2001, 29 (Pt6): 712-22). The term "inhibitor" includes both competitive and non-competitive inhibitors. Non-limiting examples of inhibitors of cystine uptake are acivicin, L-glutamate, L-serine-o-sulfate, L-cysteine ​​sulfinate, L-cysteine, L-trans-pyrrolidine-2,4-dicarboxylate, and kainite. Assays to determine whether a particular compound is an inhibitor of cysteine ​​uptake can be carried out, for example, by 35 It is an assay based on the determination of the incorporation of S-labeled cysteine.

[0130] e) Formula as described in Gumireddy et al., J Carcinog Mutagen 2013 (2013): [ka] or its disodium salt, glutathione disulfide disodium.

[0131] f) Formulas as described in Trachootham, et al., Cancer Cell 2006, 10: 241-252: [ka] Phenethyl isothiocyanate is a compound having the formula:

[0132] g) Glucocorticoid receptor antagonists, such as those described in Min, et al., J Mol Med (Berl) 2012, 90: 309-319. The term "glucocorticoid receptor antagonist" refers to a compound that binds to the glucocorticoid receptor and lacks substantial ability to activate the receptor itself. The term "glucocorticoid receptor antagonist" includes both neutral antagonists and inverse antagonists. A "neutral antagonist" is a compound that inhibits the action of an agonist but does not affect intrinsic or spontaneous receptor activity. An "inverse antagonist" is capable of inhibiting the action of an agonist at the receptor while attenuating the constitutive activity of the receptor. The term "antagonist" also includes competitive antagonists, which are drugs that bind to the same site as the natural ligand; noncompetitive antagonists, which bind to a site on the receptor that is different from the natural ligand; reversible antagonists, which bind and unbind to the receptor at a rate determined by receptor-ligand kinetics; and irreversible antagonists, which bind permanently to the receptor by forming a covalent bond with the active site or simply binding so tightly that the dissociation rate is essentially zero. Non-limiting examples of glucocorticoid receptor antagonists include RU-486 (mifepristone), RU-43044, octahydrophenanthrene, spirocyclic dihydropyridines, triphenylmethanes and diaryl ethers, chromenes, dibenzylanilines, dihydroisoquinolines, pyrimidinediones, azadecalins, arylpyrazoloazadecalins, 11-monoarylsteroids, phenanthrene, dibenzo[2.2.2]cyclooctane and derivatives ... Cycloheptanes and their derivatives, dibenzylaniline sulfonamides and their derivatives, dihetero(aryl)pentanols, chromene derivatives, azadecalins, arylquinolones, 11,21-bisarylsteroids and 11-arylsteroids and 16-hydroxysteroids, and the dual antagonist-agonists beclomethasone, betamethasone, budesonide, ciclesonide, flunisolide, fluticasone, mometasone, and triamcinolone.Whether a particular compound is a glucocorticoid receptor antagonist can be determined by commercially available kits, such as the Glucocorticoid Receptor Pathway Reporter Kit (BPS BIOSCIENCE, SAN DIEGO, CA, USA).

[0133] h) Anti-IL-6 agents such as those described in Obrador et al. J Biol Chem 2011, 286: 15716-15727. The term "anti-IL-6 agent" refers to a compound capable of reducing the activity of IL-6 by reducing the level of IL-6, completely or partially blocking its binding to its receptor, or completely or partially inhibiting its receptor activity. The term "anti-IL-6 agent" includes inhibitory antibodies against IL-6, i.e., antibodies that bind to IL-6 and prevent IL-6 from binding to its receptor, such as elcilimomab and siltuximab, and IL-6 receptor inhibitors such as tocilizumab. Assays to determine whether a particular compound is an anti-IL-6 agent include, for example, an ELISA for determining IL-6 levels, such as a kit from Life Technologies (Carlsbad, CA, USA), or an assay for determining intracellular signaling resulting from the binding of IL-6 to its receptor, such as the IL6 / STAT3 Signaling Pathway Plus PCR Array from Quiagen (Valencia, CA, USA).

[0134] i) Formula: [ka] Buthionine sulfoximine (BSO), a compound having the formula: The glutathione-depleting effect of BSO is described in Terradez P. et al., Biochem J. 1993, 292: 477-483.

[0135] j) Formula: [ka] Diethyl maleate or DEM, a compound having the formula: The glutathione-depleting effect of DEM is described in Estrela JM et al., Nat Med 1995, 1(1): 84-88.

[0136] k) Formula: [ka] NPD926 is a compound having the formula: The glutathione depletion effect of NPD926 is described in Kawamura T et al., Biochem J 2014, 463: 53-63.

[0137] l) Formula: [ka] Parthenolide is a compound having the formula: The glutathione-depleting effect of parthenolide is described in Pei S. et al., J Biol Chem 2013, 288 (47): 33542-58.

[0138] m) Formula: [ka] wherein A is C(O) or S(O)2, n=0, 1, 2 or 3, the ortho carbons of the phenyl ring are unsubstituted or substituted with halogen, R1 is selected from the group consisting of hydrogen, halogen, C≡C-alkyl, C≡C-cycloalkyl, C≡C-cycloalkyl halide, C≡C-aryl, C≡C-aryl halide and aryl groups, R2 is selected from the group consisting of hydrogen, alkyl, alkenyl and aryl groups, R3 is selected from the group consisting of hydrogen, alkyl, alkenyl and aryl groups, and R4, R5 and R6 are each independently selected from the group consisting of hydrogen, bromine, chlorine, fluorine, keto, hydroxyl, alkyl, alkenyl, alkoxy, methoxy, aminoalkyl, aminoalkenyl and aminoalkoxy groups. In particular, compounds having the formula: [ka] Piperlongumine is a compound having the formula: The glutathione-depleting effect of piperlongumine is described in Pei S. et al. (supra).

[0139] n) Inhibitors of proteins of the bromodomain and exo-terminal domain family, such as those described in Shao Q. et al., Cancer Research 2014, 74 (23):7090-102. The term "inhibitors of proteins of the bromodomain and exo-terminal (BET) domain family" or "BET inhibitors" refers to compounds that bind to the bromodomains of the bromodomain and exo-terminal (BET) proteins BRD2, BRD3, BRD4, and BRDT, and disrupt protein-protein interactions between BET proteins and acetylated histones and transcription factors. The term "BET inhibitor" includes inhibitors that target any of BRD2, BRD3, BRD4, and BRDT. Non-limiting examples of BET inhibitors include JQ1, GSK525762A, and OTX-015. An assay to determine whether a particular compound is a BET inhibitor is, for example, BioTek's (Winooski, VT, USA) Homogeneous Proximity Assay, which screens for inhibitors of BRD4.

[0140] In particular embodiments, the glutathione depleting agent of the combination of the invention is selected from the group consisting of a) Bcl-2 antisense oligodeoxynucleotides, b) inhibitors of multidrug resistance protein 1, c) inhibitors of gamma-glutamine transpeptidase, d) inhibitors of cystine uptake, e) glutathione disulfide disodium, f) phenethyl isothiocyanate, g) glucocorticoid receptor antagonists, h) anti-IL-6 agents, i) buthionine sulfoximine, j) diethyl maleate, k) NPD926, l) parthenolide, m) piperlongumine and n) inhibitors of proteins of the bromodomain and exo-terminal domain family, in particular GSK525762A or I-BET762.

[0141] In a more particular embodiment, the inhibitor of multidrug resistance protein 1 has the formula: [ka] Verapamil is a compound having the formula:

[0142] In a more particular embodiment, the inhibitor of gamma glutamine transpeptidase has the formula: [ka] The compound acivicin has the formula:

[0143] In a more particular embodiment, the inhibitor of cystine uptake has the formula: [ka] and sulfasalazine, a compound having the formula:

[0144] In a more particular embodiment, the glucocorticoid receptor antagonist has the formula: [ka] The compound is RU-486 or mifepristone, which has the formula:

[0145] In a more specific embodiment, the anti-IL-6 agent is an inhibitory antibody against IL-6 or an inhibitor of the IL-6 receptor. In a further specific embodiment, the anti-IL-6 agent is selected from the group consisting of tocilizumab, ercilimomab, and siltuximab. The term "tocilizumab" refers to a humanized monoclonal antibody against the IL-6 receptor. The term "ercilimomab" refers to a murine monoclonal antibody against IL-6. The terms "siltuximab" or "CNTO 328" refer to a chimeric monoclonal antibody against IL-6.

[0146] In a more particular embodiment, the inhibitor of the bromodomain and exoterminal domain family of proteins is selected from the group consisting of JQ1, GSK525762A and OTX-015. [ka] This refers to the compound.

[0147] The term "GSK525762A" refers to a compound of the formula: [ka] This refers to the compound.

[0148] The term "OTX-015" refers to a compound of the formula: [ka] This refers to the compound.

[0149] The term "CPI-0610" refers to the compound sold by MedKoo Biosciencies Inc. with reference catalogue number: 206117.

[0150] In certain embodiments, the glutathione depletor is diethyl maleate, GSK525762A (I-BET762), or piperlongumine.

[0151] Direct heating is stopped in step S5, and the non-ionizing AC electric field is stopped in step S6. Note that steps S5 and S6 may be performed simultaneously, or the non-ionizing AC electric field may be stopped after direct heating is stopped, such that only the non-ionizing AC electric field is applied for a predetermined period of time after direct heating is stopped.

[0152] 11 does not require any chronological order for steps S1-S5. For example, step S3 and / or step S4 (depending on whether one or both are performed in the method) may be performed simultaneously with the start of step S1, or after the start of step S1 but at a predetermined time before the start of step S2, or simultaneously with the start of step S2 or a predetermined time after the start of step S2. Furthermore, in examples where both S3 and S4 are performed, they may be performed at the same time or at different times. For example, step S3 may be performed simultaneously with the start of step S1, or after the start of step S1 but at a predetermined time before the start of step S2, or simultaneously with the start of step S2 or a predetermined time after the start of step S2, regardless of when (or whether) step S4 is performed. Meanwhile, regardless of when (or if) step S3 is performed, step S4 may be performed simultaneously with the initiation of step S1, or at a predetermined time after the initiation of step S1 but before the initiation of step S2, or simultaneously with the initiation of step S2, or at a predetermined time after the initiation of step S2. Furthermore, step S1 may be applied for a first period of time and step S2 may be applied for a second period of time, which may partially or completely overlap with the first period of time, or which may begin when the first period of time ends.

[0153] For example, a non-ionizing alternating electromagnetic field (e.g., 300 kHz) can be applied simultaneously with an anti-cancer composition and a GSH depleting agent (e.g., pterostilbene and gemcitabine). The alternating electromagnetic field is applied for two hours, and heating is applied during the two hours (e.g., heating the target site to a temperature of 52°C for 10 minutes). In another example, a non-ionizing alternating electromagnetic field (e.g., at 300 kHz) is first applied for a first period (e.g., two hours), and heating is applied in combination with the anti-cancer composition and / or GSH depleting agent after the first period has ended (e.g., pterostilbene and heating to 47°C for two hours).

[0154] In the above method, the non-ionizing alternating electromagnetic field may have a frequency of 10 kHz to 500 kHz, result in a magnetic flux density of 0.1 pT to 1 mT, or 0.1 pT to 100 μT, or 100 μT to 1 mT, and / or a corresponding electric field strength amplitude of 1 V / cm to 3 V / cm depending on tissue impedance, and may be applied for a time period of 1 minute to 24 hours.

[0155] For direct heating, it is preferred to heat the target site to a temperature of at least 42°C, preferably between 42°C and 57°C.

[0156] Targeted heating by tumor treatment area The mechanism by which the oscillating magnetic field in the tumor treatment region can cause tissue heating is by inducing Foucault currents (or "eddy currents") in the tissue. These currents rotate around the magnetic field lines in the tissue and can heat tumor cells by the Joule effect. This is due to the conductivity σ of living tissue. The conductivity of tumor tissue increases with increasing frequency of the oscillating magnetic field and is approximately 0.15 siemens / meter at 300 kHz. This conductivity provides a path for microscopic eddy currents that flow in circular paths. The power P per unit mass that heats these cells is given by the following equation: P=π 2 B 2 d 2 f 2 / (6·ρ·D)

[0157] where B is the magnetic flux density, d is the depth of the tissue to which the magnetic field is applied, f is the field frequency, ρ is the tissue resistivity (the inverse of electrical conductivity), and D is the tissue mass density.

[0158] The conductivity of tumor tissue can be up to five times higher than that of healthy tissue, with an approximate value of 0.15 Siemens / meter between 100 kHz and 300 kHz. The D of biological tissue is variable (900 kg / m 3 ~1050kg / m 3 ), D of water is 1000 kg / m 3The maximum magnetic flux density in the tumor treatment region is 1 mT. The maximum frequency is 300 kHz. In in vitro experiments, the thickness of the culture flask is approximately 1 mm, which is the value of d. This results in a P value of 20 pW / kg, which is an extremely low value. Therefore, the mechanism in the TT region is not due to heating. Cell death may be the result of charge breakdown in the mitochondria. This effect is stronger in tumor tissue due to its higher conductivity, approximately five times higher than that of healthy cells. The synergistic effect with hyperthermia may be due to the increased conductivity associated with the increased mobility of charged molecules.

[0159] All of the above is considered to form the basis of alternative embodiments in which one or more combinations of the above features are applied, fully within the scope of the present disclosure and not limited to the specific combinations disclosed above.

[0160] In light of this, there will be many alternative ways of implementing the teachings of the present disclosure. It is expected that a person skilled in the art will be able to modify and adapt the above disclosure to suit his or her own situation and requirements within the scope of the present disclosure, while retaining some or all of the technical effects of the present disclosure as disclosed or derivable from the above, in light of common general knowledge in the art. All such equivalents, modifications, or adaptations are included within the scope of the present disclosure.

Claims

1. 1. A device for treating a cancerous target site, comprising: an electromagnetic emitter including one or more applicators having an electrically insulating coating that prevents electrical contact between the applicator and the target site, the electromagnetic emitter configured to provide a tumor treatment region at the target site via the one or more applicators, the tumor treatment region being a non-ionizing alternating electromagnetic field having a frequency between 10 kHz and 300 kHz and further having a magnetic flux density between 0.1 pT and 1 mT; a heat source configured to provide heating to the target site to induce hyperthermia at the target site; an electronic controller for electronically controlling the electromagnetic emitter and the heat source; and configured to apply the non-ionizing alternating electromagnetic field and the heating independently.

2. The device described in claim 1, wherein the applicator is an electrode.

3. The device described in claim 1, wherein the applicator is a coil.

4. 10. The device of claim 1, configured to provide the non-ionizing alternating electromagnetic field to the target for a first period of time and to provide direct heating to the target region for a second period of time.

5. 5. The device of any one of claims 1 to 4, configured to apply the non-ionizing alternating electromagnetic field to the target site for a first period of time ranging from 1 minute to 24 hours.

6. 6. The device of any one of claims 1 to 5, wherein the device is further configured to provide heating to the target site for a second period of time ranging from 1 minute to 360 minutes, and optionally the heating is applied simultaneously with the non-ionizing alternating electromagnetic field for a third period of time.

7. The device of any one of claims 1 to 6, wherein the electromagnetic emitter is configured to provide an alternating electromagnetic field at the target site having a magnetic flux density of between 0.1 pT and 100 μT.

8. The device of any one of claims 1 to 6, wherein the electromagnetic emitter is configured to provide an alternating electromagnetic field at the target site having a magnetic flux density of between 0.5 μT and 1 mT.

9. The device of any one of claims 1 to 6, wherein the electromagnetic emitter is configured to provide an alternating electromagnetic field at the target site having a magnetic flux density of between 8 μT and 1 mT.

10. 10. Apparatus according to any one of the preceding claims, wherein the electromagnetic emitter is configured to provide an alternating electromagnetic field having a frequency between 100 kHz and 300 kHz.

11. The apparatus of any preceding claim, wherein the heat source is configured to heat the target site to a temperature above 42°C.

12. 12. The apparatus of any one of claims 1 to 11, wherein the heat source comprises an ultrasound emitter configured to provide ultrasound radiation at the target site, optionally the ultrasound radiation having one or more focal regions at the target site.

13. The apparatus of any preceding claim, wherein the heat source comprises an electromagnetic emitter configured to provide electromagnetic radiation to the target site.

14. 14. The apparatus of any one of claims 1 to 13, wherein the heat source comprises a fluid pump configured to pump fluid to the target site and a heater to heat the fluid before it reaches the target site.

15. The apparatus of any preceding claim, wherein the heat source comprises a conductive heat emitter configured to supply heat to the target site by thermal conduction.

Images