Method and system for applying high uniformity pulsed electric fields with a charged ring structure

Abstract

Apparatus, systems, and methods are provided for generating high uniformity pulsed electric fields. The electric fields occupy a large volume suitable for placement of a human or animal patient. An apparatus for generating electric fields is provided that includes a plurality of ring-shaped structures made of electrically conductive material, where the ring-shaped structures are charged to different voltage levels. When pulsed with electric current, the apparatus generates high uniformity electric fields in the interior region of the ring-shaped structures. These electric field pulses, when used in conjunction with a pharmacological agent, destroy cancer cells through a process called targeted osmosis lysis.

Description

[0001] Related applications

[0002] This application claims priority to U.S. Provisional Application No. 62,971,562, filed February 7, 2020, and U.S. Provisional Application No. 63 / 143,303, filed January 29, 2021, both of which are incorporated herein by reference in their entirety for all purposes.

[0003] field

[0004] This invention relates to the field of medical devices and medical treatments for diseases and physical discomfort. More specifically, this invention relates to a method and system for generating a highly uniform pulsed electric field via a charged ring structure for medical applications. Background Technology

[0005] Pulsed electric field (PEF) therapy is currently widely used in various biological and medical applications: gene delivery, electrochemical therapy, and cancer treatment. One advantage of PEF is its ability to destroy tissue or tumors in a non-thermal manner. Therefore, PEF can preserve the integrity of sensitive tissues such as blood vessels and axons. Furthermore, this non-invasive technique allows for the possibility of regeneration with healthy cells and tissues in the treatment area without leaving scars.

[0006] Traditional devices for generating pulsed electric fields consist of three parts: a pulse generator, electrodes, and the connecting links between them. The pulse generator produces square wave pulses at regular intervals. Amplitude, pulse width, period, and phase delay are the main parameters determining the shape of the output waveform. The electric field strength depends on the pulse amplitude and the distance between the electrodes and is often crucial for achieving therapeutic effects. When the electrodes are inappropriate, the intensity in a specific target area is insufficient, leading to poor therapeutic results.

[0007] Traditional techniques for generating electric fields, similar to those used in radar, have drawbacks in terms of cost and availability. A typical device using this technique is a bipolar generator, which generates short square waves with reversed polarity, partly to avoid electrode corrosion. However, a bipolar generator costs approximately twice as much as a unipolar generator. Other waveforms include exponentially decaying and sinusoidal waveforms. The sinusoidal form is slightly easier to generate because it uses a device similar to that of ordinary radio equipment, but it only reaches its peak power momentarily and therefore delivers very little energy per cycle above the critical field strength compared to a square wave.

[0008] There are currently three available techniques for generating electric field pulses. In one technique, an electric field is created between two large conductive plates, each charged to the point where a voltage difference exists between them. The patient is placed between the plates. The electric field is directed from one plate to the other and oriented perpendicular to most of the patient's body surface area, resulting in a significant reduction in the electric field within the patient. This makes controlling the field within the patient's body very difficult, as the actual field is highly sensitive to the percentage of space the patient fills between the plates. The resulting field will vary significantly with the patient's weight. The field may also differ within the patient's anatomy as local anatomical structures fill more or less of the area between the plates. For example, the field in the abdomen of a patient with a large abdomen will differ significantly from the field in the chest or head. The conductive plates can be positioned to directly contact the patient to avoid field variations. However, typical conductive plates can only contact a small portion of the patient's skin unless they are flexible.

[0009] Another technique used in laboratory experiments involves using a solenoid coil with an empty bore, within which the patient is placed. The current in the coil changes in a ramp-like manner over time, causing a change in the magnetic field, which, according to Faraday's law of induction, generates a changing electric field within the patient's body. The coil is made of a magnetic material. One drawback of this technique is the spatial variation of the electric field generated by the solenoid coil; the electric field is zero along the central axis and increases with the radius from the axis. Furthermore, if scaled up to devices the size of humans or large animals, the power requirements are extremely high, with peak power ranging from 50 to 400 kilowatts. Such high power requirements pose significant challenges to building infrastructure. This technique also requires very robust heat dissipation systems for both the device itself and the building in which it operates. The heat generated by the electrostatic ring unit can be comparable to that of other small electrical appliances such as light bulbs.

[0010] The third technique creates an electric field by sloping the magnetic field within a material with high magnetic susceptibility. The electric field generated in this way has the desired properties, but the device can be very heavy due to the need for a large amount of magnetic material. Another drawback of this technique is that, for a given pulse duration, the upper limit of the electric field strength is limited by the material properties of the magnetic material.

[0011] Despite recent advances in the use of electrical pulses to induce cell death, there remains a need in the art for improved devices and methods to destroy diseased or uncomfortable tissues, such as tumor tissue, without damaging normal tissue. In particular, there is a need for devices and methods that can generate highly uniform pulsed electric fields within large volumes. Invention Overview

[0013] In view of the foregoing, the object of the present invention is to address the need for generating highly uniform pulsed electric fields (PEFs) within large volumes for medical applications. Embodiments of the present invention relate to apparatus and methods for creating pulsed electric fields for human or animal subjects as part of a cancer treatment regimen. The present invention provides a system for generating large-volume and highly uniform electric fields suitable for placing human or animal patients inside.

[0014] An embodiment of a device for generating a pulsed electric field is described, comprising multiple coaxial conductive ring structures. The ring structures are large enough to allow a human or animal object to be placed within their interior region, separated by a distance ranging from several inches to several feet. Each ring structure is charged to a voltage level; the voltage difference between the ring structures generates an electric field in the interior region. The voltage level applied to each ring structure is designed to optimize the uniformity of the resulting electric field. According to one embodiment, the electric field is applied in the form of a series of repetitive pulses.

[0015] Another embodiment of a system is described, comprising a conductive ring structure connected to a set of driving electronics that allow a user to control the amplitude, duration, and interval of electric field pulses. The driving electronics include components for generating pulsed voltage or current waveforms, components for amplifying and filtering the output waveforms, and a microprocessor presenting a user interface for controlling the output.

[0016] The apparatus and system for generating an electric field according to the present invention have various desirable features. First, the generated electric field has high spatial uniformity. Second, the electric field is tangentially directed towards the surface of the patient lying in the apparatus. Third, the power requirement and heat generation are low compared to some other methods. Fourth, the driving electronics are relatively simple. Fifth, the device is inherently lightweight.

[0017] The electric field pulses generated by this invention, when used in conjunction with a pharmacological agent, can destroy cancer cells through a process called targeted osmotic lysis (TOL) as described in U.S. Patent No. 8,921,320, the entire disclosure of which is expressly incorporated herein by reference.

[0018] Another embodiment provides a method of therapeutic treatment via targeted permeation and dissolution, comprising administering to a human or animal subject a therapeutically effective dose of stimulation by a pulsed electric field generated by the device and system according to the invention. This method is applicable when combined with methods for blocking Na+. + ,K +- ATPases, when combined with other pharmacological agents, can be used for targeted permeation dissolution in the treatment of cancer. In some embodiments, a therapeutic treatment method by targeted permeation dissolution includes administering a therapeutically effective dose of a pulsed electric field to a human or animal subject with a tumor on a monthly basis for life or until the tumor is clinically undetectable. Brief description of the attached diagram

[0020] The invention will now be described in more detail with reference to one or more accompanying drawings illustrating exemplary embodiments.

[0021] Figure 1 Multiple ring structures are shown, which are arranged coaxially to provide an extended area of ​​electric field exposure.

[0022] Figure 2 A system for applying therapy involving an electric field is shown, the system comprising an electronic ring unit located in a housing and connected to a control system.

[0023] Figure 3 A typical pulse train associated with TOL applications is shown.

[0024] Figure 4 The stimulation duration-response curve of the pulsed electric field is shown over a day of treatment.

[0025] Figures 5A-5C Sodium channel markers of voltage-gated sodium channels (VGSCs) are shown in 4T1 allografts before and after TOL treatment.

[0026] Figure 6 The survival rate of a mouse model of triple-negative breast cancer treated with TOL is shown.

[0027] Figure 7 Survival rates after treatment in mouse models of triple-negative breast cancer treated with TOL or paclitaxel are shown.

[0028] Figures 8A-8D Sodium channel markers of VGSCs in 4T1 allogeneic grafts before and after paclitaxel treatment are shown.

[0029] Figure 9 The differences between the toroid device and the ring device in reducing tumor size in mice with ectopic 4T1 allogeneic grafts are shown.

[0030] Figure 10 The effect of digoxin dosing frequency on the effect of TOL on reducing the size of allogeneic grafts was shown.

[0031] Figure 11The effect of TOL on the growth of 4T1 allogeneic grafts in female BALBc mice that had been administered digoxin to a steady state prior to TOL treatment was demonstrated.

[0032] Figure 12 This study compares the effects of TOL on the growth of 4T1 allogeneic grafts in female BALBc mice under pretreatment with digoxin and without pretreatment.

[0033] Figure 13 The efficacy of TOL treatment was demonstrated with different treatment intervals between digoxin and PEF stimulation.

[0034] Figure 14 The growth of 4T1 allogeneic grafts in female BALBc mice receiving TOL for different durations of stimulation is shown. Detailed Implementation

[0035] It should be understood that the present invention is not limited to the specific methods, schemes, and systems described herein, and therefore can be varied. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention, which is defined only by the claims.

[0036] As used in the specification and appended claims, unless otherwise specified, the following terms shall have the meanings as follows.

[0037] As used in this article, "tumor" refers to the growth and proliferation of all proliferative cells, whether malignant or benign, as well as all precancerous lesions and cancerous cells and tissues.

[0038] “Cancer” and “cancerous” refer to or describe a physiological condition in mammals characterized by unregulated cell growth. Benign and malignant tumors, as well as latent tumors or microwound metastases, are all included in this definition.

[0039] "Subject" refers to mammals, such as but not limited to humans or non-human mammals, such as cows, horses, dogs, sheep, or cats.

[0040] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention. As used herein, the singular forms “a,” “an,” and “the” are also intended to include the plural forms unless the context clearly indicates otherwise. It will be further understood that, when used in this specification, the terms “comprises” and / or “comprising” refer to the presence of the stated features, integrals, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integrals, steps, operations, elements, components, and / or combinations thereof.

[0041] The following description and accompanying drawings fully illustrate specific embodiments to enable those skilled in the art to practice them. Other embodiments may be combined with structural, logical, electrical, process, and other variations. Parts and features of some embodiments may be included in or replace parts and features of other embodiments.

[0042] This invention addresses the need to create pulsed electric fields in large (human-sized) volumes. This need arises in the application of targeted infiltration dissolution (TOL), which utilizes such electric field pulses to stimulate the opening of sodium channels on the cell membranes of cancer cells. See U.S. Patent No. 8,921,320. A highly uniform electric field is desirable so that the associated therapeutic effect will be uniform.

[0043] The electric field is caused by, for example Figure 1 The voltage difference between the ring structures shown generates an electric field. Each ring structure has a voltage that charges it. The voltage difference between the ring structures produces an electric field between the rings, which is oriented primarily along the axis near the axis of the device. The voltage value applied to each ring structure, as well as the spatial location and diameter of the ring structures, are optimized to produce an electric field of the desired strength and uniformity.

[0044] The circular shape of the ring structure is a preferred embodiment because it produces a field with good uniformity. It should also be understood that the field can be generated using non-circular shapes, including but not limited to elliptical, polygonal, and rectangular shapes. Ring structures do not necessarily have a uniform diameter.

[0045] The ring structure can be made of conductive materials, including but not limited to metals, electrolytes, superconductors, semiconductors, plasmas, and some non-metallic materials such as graphite and conductive polymers.

[0046] By using multiple such ring structures with different voltage levels and carefully designed geometric relationships regarding their diameters, large regions with high electric field uniformity can be generated. In, for example... Figure 1In this depicted arrangement, multiple ring structures 1 are aligned so that they share a common axis and are spatially separated by a distance. In this arrangement, the patient is placed along the central axis of the device. When arranged in this way, the generated electric field extends along the axis of the ring structures; in preferred applications, this electric field will extend along the long axis of the human patient or, for many types of veterinary patients, the long axis. A larger homogeneous region can be created by increasing the number of ring structures. This design can be scaled up to an arbitrarily large number of ring structures to increase the homogeneous volume, at the cost of system cost, weight, and complexity.

[0047] To obtain an electric field pulse of a given amplitude, each ring structure is charged to a voltage level; the voltage difference between the ring structures generates an electric field in the internal region.

[0048] The device for creating the electric field can also be incorporated into a system that can be used for therapeutic purposes, such as treating certain types of cancer when combined with pharmacological agents. Specifically, the system includes one or more rings (called electrostatic ring units (ERUs)) within a housing, which are connected to a control system for applying the therapeutic effect involving the electric field. Figure 2 A block diagram of the system is shown. An electrostatic loop unit (ERU) 2 generates electric field pulses in the internal area where the patient is placed. Cable 3 connects the ERU to a drive and sensing circuit 4, which provides voltage or current pulses to the loop in the ERU 2. A sensing coil inside the ERU measures the internally generated electric field and can be used to control the output. A microprocessor 5 presents a user interface to the operator of the device and connects to the drive and sensing circuit to control the amplitude, duration, and interval of the pulses, as well as to start and stop the pulses.

[0049] The driving electronics are connected to a computer, which hosts the user interface, allowing the user to control the pulse amplitude, duration, and interval, as well as to start and stop pulse therapy. The computer can communicate with the driving electronics via a serial bus, but other options are also available.

[0050] The electric field amplitude can be controlled by an electric field sensor 4.1 in an "open-loop" arrangement or a "closed-loop" arrangement using a feedback loop. In the "open-loop" arrangement, the expected electric field output is known based on the input voltage and the current created. Feedback can take various forms, such as measuring the actual voltage applied to each loop, or measuring the applied electric field from an electric field sensor inside the device.

[0051] Voltage pulses in driving electronics can be created using many different types of amplifier configurations (4.2). Since a voltage in the drive ring is typically desired to be in the 15-100 volt range, a Class D amplifier configuration is preferred to avoid large heat dissipation in the amplifier output transistor. This configuration uses pulse width modulation (PWM) to control the amplifier output and is known for its high efficiency and low cost.

[0052] A key characteristic of the electric field generated by this invention is its high uniformity. High uniformity is ideal so that treatment is applied consistently throughout the body or treatment area. The usable treatment area for this application is a region where the field strength variation in the open space is less than about 10%.

[0053] Another important aspect of the invention is that the electric field is tangentially directed towards the surface of the patient's body while lying in the device. The desirability of a tangentially directed electric field towards the patient's body surface stems from the need to minimize the reduction of the electric field generated by polarized water molecules within the body. Water has a very high polarizability (electromagnetic susceptibility), which results in a significant reduction in the field within the body. This effect is maximized in a field perpendicular to the surface, with the reduction in electric field reaching as high as 1 / 75 to 1 / 80 of the original value. For an electric field directed along the patient's body surface, the reduction can be much smaller, ranging from almost no reduction to approximately 1 / 20 of the original value.

[0054] Another important aspect of the invention is that the device generates an electric field with very low power, resulting in low-cost drive electronics, low electrical requirements for the facility, and no impact on the HVAC system of the clinical facility. Furthermore, the device itself is lightweight.

[0055] Pulsed electric field systems can be applied to a therapeutic technique called targeted penetration dissolution (TOL). See U.S. Patent No. 8,921,320. The principle behind this technique is that pulsed electric fields stimulate the opening of sodium channels in cell membranes, allowing more sodium to enter the cell. Cancer cells are known to have more sodium channels than non-cancer cells. The therapeutic stimulation of these sodium channels by pulsed electric fields leads to an increase in sodium concentration within the cancer cells, resulting in subsequent water influx and causing the cancer cells to rupture. Normal tissue remains intact during this treatment.

[0056] Pharmacological agents used to block sodium outflow from cells, such as Na + ,K + -ATPase inhibitors can be used in conjunction with pulsed electric fields to enhance therapeutic efficacy. They can be used to block Na+. + ,K +Non-limiting examples of pharmaceutical compounds containing ATPases include: ouabain (g Strophantin); dihydroouabain; ouabainoctahydrate; ouabagenin; digoxin; digitalis; digitalis; acetyldigitoxin; acetyldigoxin; lanatoside C; deslanoside; medigoxin; gitoformate; oleanderin; oleandrin; bufotoxin; bufotoxin; bufotoxin (3,5-dihydroxy-14,15-epoxybufotoxin); palytoxin; oligomycins A, B, C, E, F, and G; rutamycin (oligomycin D); rutamycin B; strophanthin (g strophanthin). strophanthin, Acocantherine; kβ-strophanthin; strophanthidin; kstrophanthoside; Apocynum venetum glycoside; Tanshinone (Cardanolide); Chloranthidin; Peruvian glycoside; Hypothalamic Na + ,K + -ATPase inhibitor (HIF); HIF aglycone; bufotoxin; bufotoxin; marine bufotoxin; proanthocyanidins; citronellol; astragaloside A; 3,4,5,6,tetrahydroxyxanthrone; and all other Na+ + ,K + -ATPase inhibitors, combinations of each and their derivatives.

[0057] Na + ,K + -ATPase inhibitors can be delivered to a single tumor via direct or intravenous administration, to a single organ or region via intravenous or intracavitary administration, or to the whole body via intravenous, subcutaneous, intramuscular, or oral administration. Pulsed electric field stimulation of sodium channels can be delivered to a single tumor, a single organ, a part of the body, or the entire body. All types and subtypes of the VGSC family should be equally susceptible to this technology. For example, cell lines overexpressing Nav 1.1, Nav 1.2, Nav 1.3, Nav 1.4, Nav 1.5, Nav 1.5n, Nav 1.6, Nav 1.7, Nav 1.8, and Nav 1.9 are susceptible to mediated targeted lysis.

[0058] Figure 3A typical pulse train associated with TOL applications is shown. The electric field amplitude ranges from 0.1 V / m to 100 V / m in free space. The pulse consists of a positive polarization of approximately 1–50 milliseconds, followed by a negative polarization with a similar duration and amplitude. The pulse interval from end to start is 5–50 milliseconds. In applications, the precise details of timing, duration, and amplitude can vary considerably.

[0059] Figure 4 The mean size of the 4T1 allogeneic grafts observed before and after each TOL treatment is shown, with the duration of exposure to 18V / m PEF varying between 5 and 60 minutes in each of the four treatments (n=15). Figure 4 As shown, a reduction in average tumor size relative to baseline was observed when exposure was set at 30 minutes. Further increases in stimulation duration had less effect on smaller tumors. Optimal tumor reduction was observed when mice were exposed to PEF for 30 minutes or in the range of more than 15 minutes but less than 60 minutes.

[0060] Figures 5A-5C Immunohistochemical markers of voltage-gated sodium channels (VGSCs) in 4T1 allogeneic grafts before (Fig. 5A) and after (Fig. 5B) treatment with TOL alone were depicted. (DRAQ-5) TM Fluorescent probes were used to restain cell nuclei. Following TOL treatment, the number of cells highly expressing VGSCs was significantly reduced, which may explain the failure to sustain tumor reduction beyond 2 days of treatment. The low-power calibration bar in Figure 5B is 50 μm, and the high-power calibration bar in the inset is 25 μm. The histogram in Figure 5C depicts the pixel counts representing sodium channel expression observed in allogeneic grafts before and after TOL treatment. As shown in Figure 5C, TOL virtually eliminated all the most highly VGSC-expressing neoplastic cells in solid tumors. This observation can be partially explained by the loss of TOL efficacy observed after the first 2 days of treatment.

[0061] Figure 6This study describes the in vivo validation of the therapeutic efficacy of pulsed electric field (PEF)-induced permeation dissolution in a mouse model of breast cancer. Ectopic allogeneic grafts of 4T1 mouse breast cancer cells were established in immunocompetent female BALBc mice (n=12). The “TOL” group received digoxin (7 mg / kg) and was subsequently exposed to PEF generated by a ring device. This treatment was administered on day 0 (day 1 of treatment) and day 1. The “Drug-only” group received digoxin only (7 mg / kg), the “Stimulation-only” group received PMF stimulation only, and the “Vehicle” group received 10% DMSO / saline only. TOL treatment and control treatments were administered for two consecutive days (arrows). Tumor size was measured daily from day 0 (day 1 of treatment) and every other day thereafter until euthanasia was performed in accordance with NIH humanitarian endpoint criteria. Figure 4 As shown, TOL treatment significantly prolonged the post-vaccination period required to meet the humanitarian endpoint criteria compared to the control group. TOL significantly improved mouse host survival without adverse effects on behavior or tissue damage compared to the negative treatment control.

[0062] Figure 7 This study describes the in vivo validation of pulsed electric field (PEF)-induced permeation and dissolution therapy in a mouse model of breast cancer, comparing its efficacy with that of paclitaxel. Paclitaxel is currently the optimal chemotherapy regimen for treating triple-negative breast cancer. Ectopic allogeneic grafts of 4T1 mouse breast cancer cells were established in immune-active female BALBc mice (n=12) and subjected to different treatments. Figure 7 The number of days between inoculation of BALBc mice with 4T1 mouse breast cancer cells and euthanasia of the allogeneic graft meeting the humane endpoint criteria is shown. Five days post-inoculation, mice received a single intraperitoneal injection of 20 mg / kg paclitaxel (black rhombus) or an equal volume of a carrier medium for paclitaxel suspension (black inverted triangle). Csi(ξ) indicates the date of administration of paclitaxel and the paclitaxel carrier medium. An additional control group received 20 mg / kg paclitaxel on day 5 post-inoculation and then received eight doses of digoxin (Dig) at 3 mg / kg for two consecutive days (arrows) or four 30-minute-cycle PEF stimulations (Stim) (field amplitude 18 V / m, 10 ms positive / negative ramp, and 15 ms interval between stimulations) as a control for paclitaxel and TOL treatment. The allogeneic graft was measured on day 6 post-vaccination (day 0 of treatment), and again on days 1 and 2 of treatment, and then every other day thereafter until the criteria for the humane endpoint of euthanasia were met. Figure 7As shown, paclitaxel treatment had no significant effect on survival compared to the control group. Mice treated with TOL alone required significantly longer time to reach the humane endpoint for euthanasia compared to those treated with PCTX alone and the control group, but the combination of paclitaxel and TOL reduced the effect provided by TOL alone. Therefore, TOL significantly improved mouse host survival compared to mice treated with paclitaxel, which served as a positive control for triple-negative breast cancer. Concomitant treatment with TOL and paclitaxel reduced the effectiveness of TOL. The reduced effectiveness of TOL in concomitant treatment with paclitaxel may be due to the decreased VGSC expression in tumor cells caused by paclitaxel treatment, as follows: Figures 8A-8D As stated above.

[0063] Figures 8A-8D Immunohistochemical markers of VGSCs in 4T1 allogeneic grafts were depicted before paclitaxel treatment (Fig. 8A), 1 day after treatment (Fig. 8B), and 2 days after treatment (Fig. 8C). Cell nuclei were counterstained with DRAQ5. VGSC expression was markedly and progressively reduced after the initiation of paclitaxel treatment. The low-power calibration bar in Fig. 8B is 50 μm, and the high-power calibration bar in the inset of Fig. 8C is 25 μm. The histogram in Fig. 8D shows the pixel counts representing sodium channel expression observed in allogeneic grafts before and 1 day after paclitaxel treatment (Fig. 8B) and 2 days after treatment (Fig. 8C). Notably, VGSC markers decreased progressively, but the number of cells expressing the highest levels of VGSCs did not decrease. These data suggest that paclitaxel reduces VGSC expression in 4T1 mouse breast cancer allogeneic grafts. The reduction in VGSC expression may be attributed to decreased efficacy of TOL when used concurrently with paclitaxel.

[0064] Figure 9This study depicts the reduction in mean tumor area in female BALBc mice with immunogenicity from an allogeneic graft of 4T1 mouse breast cancer cells that received TOL using two types of pulsed electric field (PEF) generating devices. Mice were subjected to pulsed electric fields of varying voltage levels, generated using three stimulation devices: two helical loop designs (black bars) and one coaxial loop design (gray bars), as each device yielded the maximum field strength; 3.0 V / m and 6.0 V / m for the helical loop devices, and 36.0 V / m for the coaxial loop device. The helical loop devices used in this study are described in WO2020 / 117662. The coaxial loop device used is described herein in this application. The bar graphs summarize the mean tumor reduction observed after a single TOL treatment at different PEF intensities compared to the normalized baseline mean (white bars). It is noteworthy that when a PEF with an amplitude of 6 V / m was delivered using either a spiral-wound device or a coaxial-ring device (strip 6.0 and 6.0x) (for each stimulation group, n=8), tumor reduction was comparable when using the spiral-wound device and the coaxial-ring device. Figure 9 As shown, TOL therapy with a coaxial ring device is most effective when the field amplitude is 18V / m, and less effective when the field strength is greater or less.

[0065] Figure 10 The effect of digoxin dosing frequency on the effect of TOL in reducing the size of allogeneic grafts is shown. The figure shows the difference in mean reduction of allogeneic graft size at 4T1 after TOL treatment when neither steady-state digoxin levels were reached nor maintained. In this study, female BALBc mice received one, three, five (steady-state), or eight (maintaining steady-state) subcutaneous injections of digoxin (3 mg / kg) and four 30-minute exposures to pulsed electric field (PEF) stimulation (18 V / m, 10 ms positive / negative ramps, 15 stimulation intervals) for two consecutive days (empty arrows), totaling two hours of stimulation per hour. Figure 10 As shown, when digoxin dosing frequency resulted in steady-state drug levels below 3 mg / kg (filled triangles and rhombuses), additional exposure to PEF had little effect on tumor growth. In contrast, when digoxin reached steady-state levels (filled squares and circles), exposure to PEF led to a reduction in allogeneic graft size. Antitumor activity was enhanced when the drug was maintained at steady-state levels (filled circles). Therefore, tumor reduction in response to TOL treatment requires reaching steady-state levels of digoxin prior to PEF stimulation. Maintaining steady-state levels of digoxin throughout the stimulation period can improve efficacy.

[0066] Figure 11The effect of TOL on the growth of 4T1 allogeneic grafts is shown in female BALBc mice that were administered digoxin (3 mg / kg) to steady state (5 subcutaneous injections) for 1, 3, or 5 days prior to TOL treatment (empty arrows) (black arrows). Although growth continued in all groups (n=6), the effect of TOL on allogeneic graft growth appeared to be minimal in mice that were pretreated for only 1 day prior to TOL treatment (filled triangles). Therefore, daily pretreatment of mice with digoxin sufficient to reach steady-state levels for at least 1 day may eliminate the dose-related effectiveness of TOL in reducing tumor size.

[0067] Figure 12 The effect of TOL on the growth of 4T1 allogeneic grafts in female BALBc mice pretreated with digoxin (3 mg / kg) daily to steady state (5 subcutaneous injections) for 5 days before TOL treatment (empty arrows) (black arrows) is shown compared with the effect of TOL on the growth of allogeneic grafts in mice not pretreated with digoxin. Notably, TOL treatment in the absence of digoxin pretreatment reduced the size of allogeneic grafts by approximately 40% (filled squares), but had no effect on the growth of allogeneic grafts in digoxin-pretreated mice (filled circles). Therefore, daily pretreatment of mice with digoxin sufficient to reach steady-state levels may reduce the effectiveness of TOL treatment in reducing tumor size.

[0068] Figure 13The efficacy of TOL treatment was demonstrated at different treatment intervals between digoxin and PEF stimulation. Ectopic allogeneic grafts of 4T1 mouse breast cancer cells were established in immunocompetent female BALBc mice. These mice received five injections of digoxin (3 mg / kg) over two consecutive days to reach steady-state levels. Then, the mouse group (n=12) was treated with TOL at intervals of 0, 1, 3, 5, and 7 days after pretreatment (eight hourly subcutaneous injections of digoxin (3 mg / kg) to achieve and maintain steady state, via 4 x 30-minute exposures to pulsed electric field (PEF) stimulation (18 V / m, 10 ms positive / negative ramp, 15 stimulation intervals, for a total of 2 hours over 2 consecutive days). Digoxin pretreatment had no effect on allogeneic graft growth within 5 days. Progressive improvements in tumor reduction were observed when TOL was administered 5 and 7 days after digoxin pretreatment. A plus sign (+) indicates the day when all mice in a group reached the humane endpoint of euthanasia. Survival was prolonged in the mice group treated with TOL 5 ​​and 7 days after digoxin pretreatment in an interval-dependent manner. The data suggest that tolerance to digoxin is reversible. In small animals such as mice, a minimum of 5 days, preferably 7 days, of digoxin-free treatment is required between every 2-day treatment cycle. The digoxin-free period is approximately 2 to 4 weeks between 2-day pulsed electric field application cycles in human patients or large animals such as cats or dogs.

[0069] Figure 14 Growth of 4T1 allogeneic grafts in female BALBc mice treated with TOL for varying durations of stimulation is shown. Groups 1, 2, and 3 (n=8) received hourly injections of digoxin (3 mg / kg) on ​​day 0 to achieve and maintain steady-state drug levels, followed by exposure to PEF (18 V / m, 10 ms positive / negative ramp, 15 stimulation intervals) for 1, 2, or 3 hours. This procedure was repeated on day 4. Group 4 received eight injections of digoxin on days 0, 4, and 8 to achieve and maintain steady-state levels, followed by PEF stimulation for 4 x 30 minutes on a single treatment day. Group 5 was treated similarly, but according to the protocol, also receiving treatment for two consecutive days starting on days 0, 4, and 8. All treatment regimens showed a reduction in allogeneic graft size relative to baseline after day 1 of treatment. This response was even greater if a second treatment was administered on a subsequent day. There were no significant differences between mouse groups exposed to PEF stimulation for 1, 2, or 3 hours per day. A plus sign (+) indicates the number of days all mice in a group reached the humane endpoint of euthanasia. No significant differences were observed in mouse survival rates. The optimal safe and effective treatment regimen for TOL is to achieve and maintain steady-state digoxin levels through 2 hours of PEF stimulation for 2 consecutive days.

[0070] The electric field generated by this invention can also have other therapeutic or industrial applications.

[0071] It should be understood that the above embodiments are merely illustrative examples of numerous embodiments and variations that can constitute applications of the principles of the present invention. Other such embodiments can be readily devised by those skilled in the art without departing from the spirit or scope of the invention, and it is intended that they be considered within the scope of the invention.

[0072] Example

[0073] The following examples, including the experiments conducted and the results obtained, are provided for illustrative purposes only and should not be construed as limiting the invention.

[0074] Example 1. Large animal trials using targeted permeation and dissolution therapy

[0075] Consistent results from in vivo studies on experimental animals revealed the following: Following extensive safety testing in normal cats and dogs, experimental treatment was initiated in two dogs using the coaxial ring device of the present invention: Targeted Infiltration Dissolution (TOL) showed no adverse behavioral effects or damage to normal tissues compared to control mice, sustained reduction of xenograft size by 30-50%, and extended host mouse survival by an average of 10-14 days.

[0076] Dog 1 was a 12-year-old female Labrador Retriever with two tumors in her right lung. Chest X-rays and tumor tissue samples were obtained and immunocytochemically analyzed to determine voltage-gated sodium channel (VGSC) expression levels. VGSC expression levels were found to be sufficiently high to recommend treatment and indicated a positive response to treatment was expected. Pretreatment with digoxin was initiated to reach steady-state drug levels. On the day of treatment, the dog received an additional dose of digoxin and was then exposed to pulsed electric field (PEF) stimulation in a coaxial ring device with a field amplitude of 18 V / m. She was then sent home and returned the following day for a second stimulation cycle. The dog showed no signs of discomfort during treatment, and the owner observed no adverse cognitive or behavioral effects. Post-treatment chest X-rays showed a reduction in the size of each tumor by approximately 17–20%. Based on the initial response to treatment, a second round of treatment was administered. No adverse events occurred during treatment. An increased appetite and significantly increased activity were noted in the dog. A month later, the dog received a third round of treatment but was found to be gastrointestinal upset, lethargic, and drowsy. She was examined and samples were taken for lab tests, which revealed a moderately elevated BUN / creatinine ratio. She was given steroid injections. The dog's condition continued to deteriorate, so euthanasia was decided. Based on lab tests and clinical presentation, the sudden drop was unlikely to be related to treatment-associated tumor lysis syndrome, but rather to cancer metastasis to the brain.

[0077] Dog 2 was a 15-year-old male Labrador Retriever with two tumors in his right lung. He was unresponsive to chemotherapy. Chest X-rays were obtained, and tumor tissue samples were taken for immunocytochemistry to determine voltage-gated sodium channel (VGSC) expression levels. VGSC expression levels were found to be sufficiently high to recommend treatment and indicated that a positive response to treatment was expected. Pretreatment with digoxin was initiated to reach steady-state drug levels. On the day of treatment, the dog received an additional dose of digoxin and was then exposed to pulsed electric field (PEF) stimulation in a coaxial ring device at an amplitude of 18 V / m for 2 hours. He was then taken home and returned the following day for a second stimulation cycle. The dog showed some anxiety upon entering the transport crate but showed no signs of discomfort or adverse cognitive or behavioral effects during treatment. Post-treatment chest X-rays showed a reduction in the size of each tumor by approximately 25%. Based on the initial response to treatment, a second round of treatment was administered. No adverse events were observed during or after treatment. The tumor continued to shrink, but the amount of reduction with each treatment appeared to be slightly smaller. No significant behavioral changes were observed. The third round of treatment was administered using a smaller, bench-sized coaxial ring device. Treatment parameters were the same as before, but due to the dog's level of anxiety, a dose of acepromazine was administered before inserting it into the orifice of the device. This procedure was well tolerated. Pre-treatment X-rays were not obtained before treatment, but a comparison of the post-treatment chest X-ray after the third round of treatment with the post-treatment X-ray obtained after the second round of treatment showed a variable, but overall the tumor size decreased by approximately 5%. This finding is considered significant because the tumor was expected to grow during the period between the second and third treatments.

[0078] The dog was treated for the fourth time using a coaxial ring device of bench size, maintaining the same field strength of 18V / m for two consecutive hours over two days. Post-treatment X-ray examination revealed a stable tumor that was slightly smaller than after the third round of treatment. The dog has now completed four treatment cycles in three months, and the tumor is smaller than it was at the time of the first imaging. His owner reports that his behavior and appetite remain unchanged, and that he has experienced no serious side effects other than sedation.

[0079] In summary, these findings suggest that targeted permeation lysis could provide a safe and effective treatment for advanced cancers in large animals without compromising patients' quality of life.

[0080] Example 2. Emergency use of pulsed electric field generators in human patients

[0081] The patient has been diagnosed with refractory cervical cancer in her fifth decade of life. Her clinical problems include intractable pain, even with high doses of anesthetic on a PCA pump, and malnutrition. She is taking hydromorphone, morphine, methadone, and anxiolytics. Multiple administrations of analgesics have not provided any relief. Her ECOG performance status is 4. Her tumor is considered refractory to all standard of care, and she is deemed unsuitable for any local clinical trials. Given her extreme suffering due to tumor progression, targeted infiltration dissolution (TOL) therapy is being considered for emergency use, as her previous biopsy showed increased sodium channel expression.

[0082] The patient started taking digoxin at the following dosages: Day 1, 0.25 mg; Day 2, 0.5 mg; Day 3, 0.25 mg; Day 4, 0.25 mg; Day 5, 0.25 mg. Prior to stimulation, the patient underwent safety testing including CBC, CMP, uric acid, digoxin levels, and EKG rhythm strip measurements. The patient also received intravenous administration of allopurinol.

[0083] The patient was then placed in a coaxial ring device that provided a pulsed electric field (18 V / m field amplitude, 10 ms positive / negative ramp, and 15 ms stimulation interval). To avoid any potentially adverse interactions between the pulsed electric fields, test stimulation cycles of 15–30 seconds were initiated at 2 (minimum field strength), 4, 6, 8, 10, 12, 14, 16, and 18 V / m (treatment field strength). The patient reported no discomfort. Treatment was then provided at 18 V / m for a total of two hours, with rest periods every 15 minutes, during which blood pressure and heart rate were monitored.

[0084] Post-treatment laboratory test samples and post-treatment EKG strips were obtained. No problems were found during the post-treatment observation period. The patient was then given 1 liter of normal saline in anticipation of tumor dissolution. The patient appeared to tolerate this procedure very well.

[0085] The patient's spouse monitored the patient's blood pressure, urine output, and temperature at home. The patient reported a mild fever of 101 degrees Celsius during the night, which responded to acetaminophen treatment. The patient also experienced severe pain during the night, requiring an additional dose of the patient-breakthrough analgesia regimen. The characteristics and distribution of the pain were consistent with those reported prior to TOL treatment.

[0086] The following day, the patient returned for the second phase of the two-day protocol. Pre-treatment lab samples and EKG rhythm strips were obtained. The patient was again treated for two hours in a coaxial loop device at 18V / m, with blood pressure and heart rate checked during rest periods.

[0087] Apart from a drop in hemoglobin to 6.4 grams, the patient's laboratory results were stable. This was considered blood dilution. The patient did not receive a blood transfusion.

[0088] The patient's spouse reported that the patient experienced a fever of 101.9 degrees Celsius, which decreased to 100.7 degrees Celsius after taking acetaminophen the following evening. The patient continued to produce urine, with two measurements showing an output of 50 cc, followed by 30 cc. The patient's pain did not worsen after the second round of stimulation, and it was noted that the patient stood up and walked "in shortspurts" around the house more frequently than usual.

[0089] On the second day after treatment, the patient underwent laboratory tests that showed his hemoglobin level had returned to the patient's baseline of 7.1 grams. All other laboratory tests remained at baseline.

[0090] On the third day after treatment, the patient returned home for in-home end-of-life care. The patient was reported to be more mobile and fever-free. Pain persisted, and the dosage of anesthetic was increased. The patient was more interactive and able to engage in conversations of reasonable length. The most significant change was a marked improvement in appetite. Objective measurements of tumor density showed a decrease from 70 HU to 56 HU three days after treatment, and a further decrease to 47 HU twenty days after treatment.

[0091] The patient experienced two episodes of mild bleeding anal discharge, but reported no dizziness. The patient's blood pressure was stable at 89-102 / 60-63, and the nurse reported that the patient's complexion was good.

[0092] Various aspects of this disclosure may be implemented in one or more of the following embodiments:

[0093] Project 1): An apparatus for generating a pulsed electric field, comprising:

[0094] Multiple ring structures, each made of a conductive material, are charged to different voltage levels.

[0095] Project 2): The apparatus according to Project 1), wherein the annular structure is arranged coaxially and spatially separated.

[0096] Item 3): The apparatus according to Item 1), wherein the conductive material is selected from the group consisting of metals, electrolytes, superconductors, semiconductors, plasmas, graphite and conductive polymers.

[0097] Item 4): The apparatus according to Item 1), wherein the annular structure is circular in shape.

[0098] Item 5): The apparatus according to Item 1), wherein the annular structure is non-circular in shape.

[0099] Item 6): The apparatus according to Item 4), wherein the annular structures have the same diameter.

[0100] Item 7): The apparatus according to Item 4), wherein the annular structure has a different diameter.

[0101] Item 8): The apparatus according to Item 4), wherein the diameter of the annular structure is large enough to place a human or animal object within the annular structure.

[0102] Item 9): The apparatus according to Item 1), wherein a human or animal object is placed along the central axis of the apparatus.

[0103] Item 10): The apparatus according to Item 1), wherein the annular structure is separated at a distance ranging from a few inches to a few feet.

[0104] Item 11): The apparatus according to Item 1), wherein the pulsed electric field is created by applying different voltage levels to the ring structure.

[0105] Item 12): The apparatus according to Item 1), wherein the voltage level applied to each ring structure is configured to optimize the uniformity of the pulsed electric field.

[0106] Item 13): A system for generating a pulsed electric field, the system comprising the means according to any one of items 1)-12), the system further comprising:

[0107] Drive and sensing circuits,

[0108] Multiple cables connect the device to the drive and sensing circuitry, and

[0109] A microprocessor that provides a user interface for operating the device and the driving and sensing circuitry.

[0110] Item 14): A method for therapeutic treatment by targeted permeation dissolution, the method comprising administering to a human or animal subject in need a therapeutically effective dose of a pulsed electric field generated by the device according to Items 1)-12).

[0111] Project 15): According to the method described in Project 14), wherein the therapeutically effective dose of the pulsed electric field is 2 hours for two consecutive days with a field amplitude of 18V / m.

[0112] Item 16): The method according to Item 15) further includes administering a therapeutically effective dose of the pulsed electric field to a human or animal subject with a tumor on a monthly basis until the tumor is no longer clinically detectable.

[0113] Item 17): The method according to Item 15) further includes administering a therapeutically effective dose of the pulsed electric field to a person or animal subject with a tumor on a monthly basis throughout life.

[0114] Item 18): The method according to Item 14) further includes administering an anti-Na+ blocker to the human or animal subject. + ,K + -The effective dose of pharmacological agents for the treatment of ATPase.

[0115] Project 19): According to the method described in Project 18), wherein, for blocking Na + ,K + The pharmacological agent for ATPase is digoxin.

[0116] Project 20): The method according to Project 19), wherein a steady-state level of digoxin is obtained in the human or animal subject before the pulsed electric field is applied.

[0117] Project 21): The method described in Project 20) wherein the steady-state level of digoxin in mice is achieved at a dose of 3 mg / kg per hour.

[0118] Project 22): According to the method described in Project 19), there is a period without degaussin between the application of the pulsed electric field every 2 days.

[0119] Project 23): According to the method of Project 22), wherein the period without digoxin between the application of the pulsed electric field every 2 days is at least 5 days.

[0120] Project 24): According to the method of Project 22), the period without digoxin between the application of the pulsed electric field every 2 days is approximately 2 to 4 weeks.

[0121] List of reference numerals

[0122] 1. Ring structure

[0123] 2 electrostatic ring units

[0124] 3 cables

[0125] 4. Drive and Sensing Circuits

[0126] 5 microprocessors.

Claims

1. An apparatus for generating a pulsed electric field, comprising: An electrostatic ring unit, which is connected to a driving and sensing circuit, and Multiple ring structures are positioned along a longitudinal axis, wherein the ring structures are made of a conductive material, and wherein the driving and sensing circuits are configured to charge the ring structures to different voltage levels.

2. The apparatus according to claim 1, wherein, The ring structures are arranged coaxially and spatially separated.

3. The apparatus according to claim 1, wherein, The conductive material is selected from the group consisting of: metals, electrolytes, superconductors, semiconductors, plasmas, graphite, and conductive polymers.

4. The apparatus according to claim 1, wherein, The ring structure is circular in shape.

5. The apparatus according to claim 1, wherein, The ring structure is not circular in shape.

6. The apparatus according to claim 4, wherein, The ring structures have the same diameter.

7. The apparatus according to claim 4, wherein, The ring structures have different diameters.

8. The apparatus according to claim 4, wherein, The ring structure is configured to allow objects to be placed within it.

9. The apparatus according to claim 1, wherein, The electrostatic ring unit is configured to allow objects to be placed along their longitudinal axis.

10. The apparatus according to claim 1, wherein, The ring structures are separated by a distance ranging from a few inches to a few feet.

11. The apparatus according to claim 1, wherein, The pulsed electric field is created by applying different voltage levels to the ring structure.

12. The apparatus according to claim 1, wherein, The voltage level applied to each ring structure is configured to create a uniform pulsed electric field.

13. A system for generating a pulsed electric field, the system comprising the means according to any one of claims 1-12, the system further comprising: Drive and sensing circuits, Multiple cables connect the device to the drive and sensing circuitry, and A microprocessor that provides a user interface for operating the device and the driving and sensing circuitry.

14. A system for a therapeutic treatment involving targeted permeation and dissolution, the system comprising means according to any one of claims 1-12, wherein the system is configured to apply a pulsed electric field generated by the means to a subject.

15. The system according to claim 14, wherein, The system is also configured to apply the pulsed electric field to subjects with tumors monthly until the tumor is no longer clinically detectable.

16. The system according to claim 14, wherein, The system is also configured to apply the pulsed electric field to the subject with a tumor on a monthly basis throughout the subject's lifetime.

17. The system according to claim 14, wherein, The system is also configured to apply a blocking agent to the target. + ,K + - Pharmacological agents of ATPase.

18. The system according to claim 17, wherein, Used to block Na + ,K + The pharmacological agent for -ATPase is digoxin.

19. The system according to claim 18, wherein, The system is also configured to obtain a steady-state level of digoxin in a human or animal subject prior to the application of the pulsed electric field.

20. The system according to claim 19, wherein, The steady-state level of digoxin in mice was achieved at a dose of 3 mg / kg per hour.

21. The system according to claim 18, wherein, The system is also configured such that there is a period without degaussian between the application of the pulsed electric field every two days.

22. The system according to claim 21, wherein, The period without digoxin, between the application of the pulsed electric field every 2 days, is at least 5 days.

23. The system according to claim 21, wherein, The period without digoxin, between the application of pulsed electric fields every 2 days, lasts 2 to 4 weeks.

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