A System And Method For Dynamic Electroporation To Accelerate Delivery Of Biomolecules
Abstract:
The present disclosure relates to a system and a method for performing dynamic electroporation to accelerate the delivery of biomolecules within cells at a target site of a subject. In one aspect, a system for performing the dynamic electroporation comprising a handheld electroporation probe and a pulse management system is provided. The system comprises an electrode assembly releasably mounted at the proximal end of the probe which comprises a plurality of high-frequency switchable electrodes having dual modes of operation. In another aspect, an improved method for performing dynamic electroporation is provided which assures that the required amount of biomolecules are entrapped within the cells at the target site in a short period of time with minimal or no cell damage. The invention overcomes previous problems of assuring delivery of effective amount of biomolecules inside the cells of the target site in each subject.
Get Free WhatsApp Updates!
Notices, Deadlines & Correspondence
E-2 GIDC Electronic Estate sector 26, Gandhinagar -382028 Gujarat, India
Inventors
1. Arvindkumar L. Patel
E-2 GIDC Electronic Estate sector 26, Gandhinagar -382028 Gujarat, India
Specification
Claims:Claims
We Claim:
1. A dynamic electroporation system (10) for accelerating delivery of biomolecules into cells of a target site of a subject comprising
a handheld electroporation probe (100) comprising
a cylindrical housing (101) having a proximal end (101a) and a distal end (101b);
an electrode assembly (102) releasably mounted at the proximal end (101a) of the housing, wherein the electrode assembly comprises
an electrode connector (102d) having a first end (102d-i) having first diameter (D1) and a second end (102d-ii) having second diameter (D2), wherein the first diameter (D1) is greater than the second diameter (D2) of the second end, wherein the second end of the electrode connector has a central projection (p) and peripheral ring (r) surrounding the central projection (p) in a manner that form an annular gap (g) between the central projection (p) and the peripheral ring (r),
at least one first electrode (102a) mounted on the central projection (p) of the electrode connector (102d), wherein the first electrode comprises a plurality of embossed sub-electrodes (102c), and
at least one second electrode (102b) mounted on the peripheral ring of the electrode connector (102d),
a pulse generating means (104) to supply a plurality of dynamic pulses of energy to the electrode assembly; and,
a pulse management system (200) configured to schedule delivery of the plurality of dynamic pulses of energy based on at least one of a subject information, an electrical charge of the biomolecule and an electrical charge measured at the target site.
2. The system according to claim 1 wherein the pulse management system (200) is configured to schedule delivery of the plurality of pulses of energy by regulating at least one of frequency, strength and duration of the pulses of energy delivered at the target site.
3. The system according to claim 1, wherein the first electrode (102a) and the second electrode (102b) are high frequency switchable electrode having dual mode of operation, wherein
in a first mode of operation, the first electrode (102a) and the second electrode (102b) are configured to perform sensing of electrical charge present at the target site, and,
in a second mode of operation, the first electrode (102a) and the second electrode (102b) are configured to deliver a plurality of dynamic pulses of energy at the target site.
4. The system according to claim 3, wherein the first electrode (102a) and the second electrode (102b) are made up of gold or gold plated material.
5. The system according to claim 1, wherein the first electrode (102a) are configured to deliver positively charged pulses of energy and the second electrode (102b) are configured to deliver negatively charged pulses of energy at the target site.
6. The system according to claim 1, wherein each of the first electrode (102a) and the second electrode (102b) has an inverted tri-podal structure, a ring structure or an annular structure.
7. The system according to claim 1, wherein the electroporation probe (100) further comprises an electrode assembly retaining and releasing means (103) to selectively release the electrode assembly (102) from the proximal end (101a) of the housing (101).
8. The system according to claim 1, wherein the electrode assembly (102) further comprises a LED indicator (106) to indicate at least one of a mode of operation of the first electrode (101a) and the second electrode (101b).
9. The system according to claim 1, wherein the pulse management system (200) uses artificial intelligence to schedule the delivery of the plurality of dynamic pulses of energy based on at least one of subject information, electrical charge of the biomolecule and an electrical charge measured at the target site.
10. The system according to claim 1, wherein the pulse management system (200) is adapted to be accommodated within the hand-held electroporation probe (100).
11. The system according to claim 1, wherein the pulse management system (200) is operatively connected with the hand-held electroporation probe (100) by wired or wireless connection means.
12. The system according to claim 1, wherein the first electrode (102a) and the second electrode (102b) are configured to deliver the plurality of dynamic pulses of energy in total amount ranging from 1 mJ to 180 mJ.
13. A method (80) for dynamic electroporation for accelerating delivery of biomolecules into cells of a target site of a subject comprising
a) sensing (801) a base electrical charge present at the target site;
b) comparing (802) the base electrical charge measured with a first threshold, wherein the first threshold is determined based on a subject threshold, wherein the first threshold is determined based on the subject information and a first charge information;
c) delivering (803) a first set of plurality of dynamic pulses of energy at the target site, if the base electrical charge is greater than the first threshold;
d) sensing (804) a residual electrical charge present at the target site after delivery of the first set of dynamic pulses of energy;
e) comparing (805) the residual electrical charge measured with a second threshold, wherein the second threshold is determined based on the subject information and a second charge information;
f) delivering (806) a second set of plurality of dynamic pulses of energy at the target site, if the residual electrical charge measured is greater than the second threshold; and,
g) iteratively repeating (807) step c) to step f) until the residual electrical charge measured at the target site is lesser than the second threshold, wherein the delivery of the plurality of dynamic pulses of energy are scheduled based upon at least one of a subject information, an electrical charge of the biomolecule and the electrical charge measured at the target site.
14. The method according to claim 13 further comprises terminating (809, 808) the process, if the base electrical charge measured at the target site is lesser than the first threshold, and if the residual electrical charge measured at the target site is lesser than the second threshold.
15. The method according to claim 13, wherein the first charge information indicates the total amount of biomolecules administered at the target site, while the second charge information indicates the total number of biomolecules that needs to be administered or entrapped inside the cells at the target site.
16. The system according to claim 1 and the method according to claim 13, wherein the subject information comprises information related to at least one of age, gender, race, origin, a medical history, an anatomy, a habitat, an environmental and occupational information of the subject.
17. The method according to claim 13, wherein the total time period for completing one iteration is 10 µs to 200 ms and a total time period between two consecutive iterations is 250 ms.
18. The method according to claim 13, wherein the method is a closed loop process.
19. The system according to claim 1 and method according to claim 13, wherein the target site is intra-dermal site.
, Description:1. Title of the Invention
A SYSTEM AND METHOD FOR DYNAMIC ELECTROPORATION TO ACCELERATE DELIVERY OF BIOMOLECULES
2
Name Nationality Address
ALP MULTI-TECH PVT. LTD Indian E-2 GIDC Electronic Estate sector 26, Gandhinagar -382028, Gujarat, India
3.Preambletothedescription
The following specification particularly describes the invention and the manner in which it is to be performed.
FIELD OF INVENTION:
[001] The present disclosure relates generally to a dynamic electroporation device, which dynamically customize an electric field-induced trans-membrane potential for each subject to achieve efficient electroporation in all subjects.
BACKGROUND OF THE INVENTION:
[002] Use of electroporation techniques to deliver various biomolecules such as protein, vaccine, DNA, antigen, etc. is practiced since a long time. Electroporation ("EP") is a biotechnological process of increasing cell permeability by creating temporary pores in the cell membrane, through which biomolecules can travel inside the cells. Here, the temporary pores are created by applying electrical pulse at a predetermined voltage lasting through few microseconds to a millisecond which disturbs the phospholipid bilayer of the membrane and results in the formation of temporary pores.
[003] Electroporation has a significant impact on vaccine immunogenicity and efficacy because of its potential to increase the antigen delivery by several folds and improves response rates over the delivery by direct injection of DNA. Electroporation is an efficient technique for cell poration because the parameter in this system can be easily controlled and optimized for efficient transfection and positive immune response. Electric parameters, temperature, pH of suspending medium are known to influence the efficiency of the pulsed electric field (PEF) induced DNA transfection of cells. PEF-induced transfection has been used as a model system to establish a quantitative relationship between these parameters and transfection efficiency.
[004] Electroporation is a threshold-dependent phenomenon in which the electric field intensity must be higher than the critical threshold to induce cell permeability. To achieve success, the electrical energy must induce sufficient trans-membrane voltage for sufficient time duration to induce membrane permeability and also not exceeding the upper limits of the same to prevent cell death or to destroy or inactivate the biomolecule.
[005] The lipid bilayer membrane of a cell is a simple capacitor that stores a charge and acts as a di-electric between the highly charge conductive intracellular and extracellular environments. The electric field creates dipoles of molecules from protein and carbohydrates, which causes to orient themselves with respect to the field. It will distribute within and around the cell such that the side of the cell facing the cathode is depolarized, and the other facing the anode is hyperpolarized due to the differential accumulation of charge on either side of the plasma membrane. The amount of lipid present in the subject inherently affects external electric force that needs to be applied to generate an effective trans-membrane potential at the target site.
[006] Once this electric field induced trans-membrane potential exceeds the dielectric strength of the membrane, the membrane undergoes a di-electric breakdown resulting in the permeation events ultimately resulting in the formation of the electro pores that allows the electro-permeability of water and limited ion flow.
[007] Traditional electroporation technique is less successful for the delivery of large biomolecules as larger biomolecules often failed to travel inside target cells due to the creation of smaller and temporary pores when pulses are delivered for a short period of time. Increase in exposure to the electric field results in pore stabilization and which results in larger pores that allow entry and exit of the larger impermeable molecules like DNA, vaccine & various macromolecules to migrate in and out of the cell.
[008] Membrane destabilization results in the formation of pores and permeation events, the process starts with the formation of small pores then coalesce into larger pores. Majority of the larger pores likely close faster than the smaller pores, in an exponential manner. Thus, if the DNA is not present at the time of pulsing, all the pores permeable to DNA will close till the time of DNA addition.
[009] Most of available electroporation devices are conceptually based on the constant voltage and constant current systems, utilizing a predetermined voltage between the electrodes or by utilizing a constant current at specific predetermined load impedance. However, the unregulated current may generate some amounts of heat in the tissue that can easily lead to cell damage or cell death. Prior devices incorporate feedback means or controlling means which continuously monitor the current between the electrodes and compare it to a pre-set current, which is the same for every subject, and continuously make energy-output adjustments to maintain the monitored current at pre-set levels.
[0010] Here, practices of maintenance of current at pre-set levels sufficiently resolve the problem that the electric current delivered will not produce any adverse side effect or cause cell death. However, it does not guarantee that the delivered current was sufficient to create temporary pores of sufficient size to assure transfection of the larger biomolecules within the target cells in each subject. Different subject has a different response to the external electrical force due to various parameters. For example, depending upon the constitution of the skin, especially cell membrane of the skin cells, they might require a low or high amount of current to create temporary electro-pores. In other words, the currently used practices to use same optimized current for all the subjects do not guarantee efficient electroporation for delivery of larger biomolecules as the same current cannot ideally represent the cell constitution of all the subjects.
[0011] Besides, several hospitals are involved in the fraudulent practices of delivering placebo or saline solution to the subject instead of the active biomolecules. The subject or patient, who pays for the delivery of active biomolecules, receives only placebo/saline solution or solution with expired or inactive biomolecules or a lesser number of biomolecules. Here, the patient does not provide with any measures to check the constitution of the solution to ensure that he is actually administered with a sufficient quantity of the active biomolecules.
[0012] Hence, there is an urgent need to develop an electroporation device that solves at least all of the above problems.
OBJECTS OF THE INVENTION:
[0013] Some of the objects of the system of the present disclosure which at least one embodiment herein satisfies are as follows: It is an object of the present disclosure to ameliorate one or more problems of the prior art or to at least provide a useful alternative.
• An object of the present disclosure is to provide a dynamic electroporation system that dynamically determines the required electric field-induced trans-membrane potential for generating optimum electro-pores in each subject.
• Another object of the present disclosure is to provide a dynamic electroporation system that dynamically schedules the delivery of the multiple pulses of energy to generate required electric field-induced trans-membrane potential as predetermined by the system.
• Yet another object of the present disclosure is to provide a dynamic electroporation system that customizes electric field-induced trans-membrane potential for each subject which is sufficient for the successful delivery of the biomolecules inside the cells of the target site of each subject.
• Yet another object of the present disclosure is to provide a dynamic electroporation system that customizes electric field-induced trans-membrane potential for each subject considering the load impedance and the charge of the biomolecules that needs to be delivered.
• Still another object of the present disclosure is to develop a dynamic electroporation system that provides a non-invasive and painless delivery of the biomolecules within the subjects.
• Another object of the present disclosure is to provide an electroporation device that enables a subject to ensure the administration of active biomolecules by himself.
SUMMARY OF THE INVENTION
[0014] In one embodiment of the present invention, a dynamic electroporation system for accelerating the delivery of biomolecules into cells of a target site of a subject is provided, comprising a handheld electroporation probe and a pulse management system.
[0015] The handheld electroporation probe comprises a cylindrical housing having a proximal end and a distal end, an electrode assembly releasably mounted at the proximal end of the housing, and pulse generating means to supply a plurality of dynamic pulses of energy to the electrode assembly. The electrode assembly comprises an electrode connector having a first end having first diameter and a second end having the second diameter, wherein the first diameter is greater than the second diameter of the second end, wherein the second end of the electrode connector has a central projection and peripheral ring surrounding the central projection in a manner that form an annular gap between the central projection and the peripheral ring; at least one first electrode mounted on the central projection of the electrode connector, wherein the first electrode comprises a plurality of embossed sub-electrodes, and at least one second electrode mounted on the peripheral ring of the electrode connector.
[0016] Also in one embodiment of the invention, the pulse management system is configured to schedule delivery of the plurality of dynamic pulses of energy based on at least one of subject information, an electrical charge of the biomolecule, and an electrical charge measured at the target site. The pulse management system is further configured to schedule delivery of the plurality of pulses of energy by regulating at least one of frequency, strength, and duration of the pulses of energy delivered at the target site.
[0017] In a preferred embodiment of the invention, the first electrodes and the second electrodes are high frequency switchable electrodes having dual mode of operation, In the first mode of operation, the first electrode and the second electrode are configured to perform sensing of electrical charge present at the target site, and, in the second mode of operation, the first electrode and the second electrode are configured to deliver a plurality of dynamic pulses of energy at the target site. In a preferred embodiment of the invention, the first electrode and the second electrode are made up of gold or gold plated material.
[0018] Also in one embodiment of the invention, the first electrode is configured to deliver positively charged pulses of energy and the second electrode are configured to deliver negatively charged pulses of energy at the target site. Further, each of the first electrodes and the second electrodes may have an inverted tri-podal structure, a ring structure or an annular structure.
[0019] Preferably, the electroporation probe further comprises an electrode assembly retaining and releasing means to selectively release the electrode assembly from the proximal end (101a) of the housing (101). In an embodiment of the invention, the electrode assembly includes a LED indicator to indicate at least one mode of operation of the first electrode (101a) and the second electrode (101b).
[0020] In a preferred embodiment of the invention, the pulse management system is configured to use artificial intelligence to schedule the delivery of the plurality of dynamic pulses of energy based on at least one of subject information, the electrical charge of the biomolecule and an electrical charge measured at the target site.
[0021] In a second aspect of the present invention, a method for dynamic electroporation for accelerating the delivery of biomolecules into cells of a target site of a subject is provided. The method includes following steps:
a) sensing a base electrical charge present at the target site;
b) comparing the base electrical charge measured with a first threshold, wherein the first threshold is determined based on a subject threshold, wherein the first threshold is determined based on the subject information and a first charge information;
c) delivering a first set of a plurality of dynamic pulses of energy at the target site, if the base electrical charge is greater than the first threshold;
d) sensing a residual electrical charge present at the target site after delivery of the first set of dynamic pulses of energy;
e) comparing the residual electrical charge measured with a second threshold, wherein the second threshold is determined based on the subject information and a second charge information;
f) delivering a second set of a plurality of dynamic pulses of energy at the target site, if the residual electrical charge measured is greater than the second threshold; and,
g) iteratively repeating step c) to step f) until the residual electrical charge measured at the target site is lesser than the second threshold, wherein the delivery of the plurality of dynamic pulses of energy are scheduled based upon at least one of a subject information, an electrical charge of the biomolecule and the electrical charge measured at the target site.
[0022] The method further comprises step of terminating the process, if the base electrical charge measured at the target site is lesser than the first threshold, and if the residual electrical charge measured at the target site is lesser than the second threshold. In a preferred embodiment of the invention, the first charge information indicates the total amount of biomolecules administered at the target site, while the second charge information indicates the total number of biomolecules that needs to be administered or entrapped inside the cells at the target site.
[0023] In one embodiment of the invention, the subject information comprises information related to at least one of age, gender, race, origin, medical history, anatomy, habitat, environmental and occupational information of the subject.
[0024] The foregoing has outlined rather broadly the technical features of the present disclosure so that those skilled in the art may better understand the detailed description that follows. Additional features and advantages of the disclosure will be described hereinafter that form the subject of the claims. Those skilled in the art will appreciate that they may readily use the conception and the specific embodiments disclosed as a basis for modifying or designing other structures for carrying out the same purposes of the present disclosure. Those skilled in the art will also realize that such equivalent constructions do not depart from the spirit and scope of the disclosure in its broadest form.
BRIEF DESCRIPTION OF DRAWINGS
[0025] The embodiments herein will be better understood from the following detailed description with reference to the accompanying drawings, in which:
[0026] FIG. 1 is a schematic diagram of a dynamic electroporation system for accelerating delivery of biomolecules into cells of a target site of a subject in accordance with an embodiment of the present disclosure.
[0027] FIG. 2 is a front view of a handheld electroporation probe in accordance with an embodiment of the present disclosure.
[0028] FIG. 3a is an exploded view of a handheld electroporation probe in accordance with an embodiment of the present disclosure.
[0029] FIG. 3b is a perspective view of a handheld electroporation probe from the top, front, side and bottom in accordance with an embodiment of the present disclosure.
[0030] FIG. 4 is an isometric view of an electrode assembly of the electroporation probe in accordance with an embodiment of the present disclosure.
[0031] FIG. 5 is an enlarged isometric view of an electrode assembly in an assembled state in accordance with an embodiment of the present disclosure.
[0032] FIG. 6 is an exploded view of an electrode assembly in a dis-assembled state in accordance with an embodiment of the present disclosure.
[0033] FIG. 7 is a perspective view of electrode assembly from the front and bottom view in accordance with an embodiment of the present disclosure.
[0034] FIG. 8 flow chart illustrating a method of performing dynamic electroporation for accelerating delivery of biomolecules into a target site in accordance with an embodiment of the present disclosure.
DEFINITION
[0035] The embodiments herein and the various features and advantageous details thereof are explained more fully with reference to the non-limiting embodiments that are illustrated in the accompanying drawings and detailed in the following description. Descriptions of well-known components and processing techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be construed as limiting the scope of the embodiments herein.
[0036] It will be readily understood that the components of the embodiments as generally described herein and illustrated in the appended figures could be arranged and designed in a wide variety of different configurations. Thus, the following more detailed description of various embodiments, as represented in the figures, is not intended to limit the scope of the present disclosure, but is merely representative of various embodiments. While the various aspects of the embodiments are presented in drawings, the drawings are not necessarily drawn to scale unless specifically indicated.
[0037] The present disclosure may be embodied in other specific forms without departing from its spirit or essential characteristics. The described embodiments are to be considered in all respects as illustrative. The scope of the disclosure is, therefore, indicated by the appended claims.
[0038] Reference throughout this specification to features, advantages, or similar language does not imply that all of the features and advantages that may be realized with the present disclosure should be or are in any single embodiment of the disclosure. Furthermore, the described features, advantages and characteristics of the disclosure may be combined in any suitable manner in one or more embodiments. One skilled in the relevant art will recognize, in light of the description herein, that the disclosure can be practiced without one or more of the specific features or advantages of a particular embodiment. In other instances, additional features and advantages may be recognized in certain embodiments that may not be present in all embodiments of the disclosure.
[0039] Reference throughout this specification to “one embodiment,” “an embodiment,” or similar language means that a particular feature, structure, or characteristic described in connection with the indicated embodiment is included in at least one embodiment of the present disclosure. Thus, the phrases “in one embodiment”, “in an embodiment”, and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
[0040] As used herein, unless otherwise specified the use of the ordinal adjectives “first”, “second”, “third”, etc., to describe a common object, merely indicate that different instances of like objects are being referred to, and are not intended to imply that the objects so described must be in a given sequence, either temporally, spatially, in ranking, or in any other manner.
[0041] As used herein, the singular form “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. As used in this document, the term “comprising” means “including, but not limited to”.
[0042] Reference throughout this specification to the terms “distal” and “proximal” are used in reference to the position of the hand held electroporation probe relative to a target site of a subject when held by a user.
Detailed Description
[0043] Various embodiments disclosed herein provide a dynamic electroporation system and method for accelerating delivery of biomolecules within a cells of target site of a subject. In a preferred embodiment of the present invention, the subject includes mammals, more preferably human. In an embodiment of the invention biomolecules needs to be delivered in the target site are macromolecules, preferably DNA, Protein, vaccines, RNA, drugs etc.,
[0044] The electroporation process depends upon cell impedance, charge voltage and electro-permeabilization process timing. These parameters significantly vary from one subject to another subject due to variety of factors.
[0045] For example, the cell impedance is depends upon the composition of the tissue or cells, especially cell membrane. The cell or tissue composition is affected by various factor which includes, but not limited to, inheritance, living patterns, personal habits, lifestyle, exposure to certain pollutant, chemical or radical (different for different profession or environment where the subject work or lives), location, age, gender, race, origin, medical history etc., For example, a man has larger muscular mass then the women. A person who consumes more fatty or oily food will have a larger lipid deposit than the person who consumes less fatty or oily food.
[0046] These factors significantly vary the cell impedance in different subjects. As the cell impedance is varied, so an amount of external electrical charge that is required to generate effective trans-membrane potential in each subject is varied among different subject. In other words, an external electrical charge required to be delivered at a target site to generate effective trans-membrane potential is different in children compared to external electrical charge required for generating similar effective trans-membrane potential in adult. Similarly, a person with larger muscle mass or fat mass would require higher amount of the external electrical charge then the person with lower muscular or fat mass or vice versa. Hence, in order to develop an effective trans-membrane potential required to carry out an effective electroporation at a target site, all such parameter that possibly affects the impedance of the cells needs to be considered.
[0047] Additionally, each biomolecules carries more or less charge with themselves. When such biomolecules are delivered at the target site, the charge carried by the biomolecules significantly affects the electrical field present at the target site. Additionally, prior knowledge of such electrical charge can be used to quantify a total number of the biomolecules actually present at a target site.
[0048] Prior systems were based on an assumption that each cell has similar response to same amount of external electrical force. But in reality, each cell of a target site has different response to the external electrical force based on their impedance or composition/constituents. This may significantly affect the formation of the electropores through which biomolecule travel inside the cells. Success of electroporation process is heavily relied on how a cell responds to the external electrical force. Hence, consideration of the actual response of target cells is crucial factor to achieve successful electroporation. ¬¬¬¬All these parameters that affect electrical field at the target site must be taken into consideration to determine electroporation parameters such that an effective trans-membrane potential is developed at a target site of one subject.
[0049] The present invention relies on generating an customized effective trans-membrane potential for the subject such that efficient delivery of large number of biomolecules inside the cells of the target site is achieved in short period of time, that is also by applying lesser amount of external electrical force compared to prior art.
[0050] The external electrical force is applied at the target site by delivering multiple of pulses of energy with specific intensity for specific duration. The delivery pattern of the pulses of energy generates a trans-membrane potential at the target site which needs to be sufficient to produce larger pores through which the larger biomolecules can travel inside the cells of target site. Hence, the delivery pattern of the pulses of energy needs to be modified for each subject depends upon their cell constitution or load impedance and the charge carried by the biomolecules. The delivery patterns of pulses of energy can be modified by modifying duration and intensity of the pulses of the energy for each subject. However, the change in the delivery patterns of pulses of energy must be in accordance with the effective trans-membrane potential that needs to be produced.
[0051] Various embodiments disclosed herein provide a system and a method for performing a dynamic electroporation for accelerating delivery of biomolecules into a target site of a subject.
[0052] FIG. 1 is a schematic diagram of a dynamic electroporation system 10 for accelerating delivery of biomolecules into cells of the target site of a subject according to an embodiment of the present invention. The dynamic electroporation system 10 comprises a handheld electroporation probe 100 (referred hereinafter as “EP probe” for brevity) and a pulse management system 200. In one embodiment of the present invention, the EP probe 100 is adapted to accommodate the pulse management system 200. In another embodiment of the present invention, the EP probe 100 and the pulse management system 200 are operatively connected via wired or wireless connection. That is, the EP probe 100 and the pulse management system 200 are connected by a power supplying cable 300 or wireless connection such as Bluetooth, IR, Wi-Fi, etc.
[0053] The pulse management system 200 comprises a user interface 201, a database 202, a processor 203, and a power supplying means 204. User interface 201 can include an alphanumeric keypad, touch screen, computer mouse, push-buttons and/or toggle switches, or another suitable component to receive input from a human user or operator. The user interface 201 can also include a CRT screen, LED screen, LCD screen, liquid crystal display, printer, display panel, audio speaker, or another suitable component to convey data to a human user. The power supplying means 204 can be a cable that powers the pulse management system 200.
[0054] The Databases 200 may include one or more computing devices configured with appropriate software to perform operations consistent with training as well as generation of a delivery pattern of pulses of energy depend upon various parameters such as subject information, cell impedance, and electrical charge of biomolecules. The database 200 can be automatically updated over time by performing real-time electroporation with various subjects.
[0055] The processor 203 is configured to train an artificial intelligence based network or database with various factors affecting the electroporation process parameter. Such factors include, but are not limited to, a subject information, cell impedance, cell composition, electrical charge of biomolecules, and delivery pattern of pulses of energy and their interrelation;
[0056] The pulse management system 200 includes a computer, mobile, tablet, a digital or analog processing apparatus, programmable logic array, a hardwired logic circuit, an application specific integrated circuit (“ASIC”), or other suitable devices. The pulse management system 200 has a user interface that enables the user to interact with the pulse management system, i.e., to input the data or to view the operating parameter of the EP probe 100. Alternatively, when the pulse management system 200 is incorporated inside the EP probe 100, the EP probe can be configured to include a displaying means that display operation or process parameters.
[0057] FIG. 2 is a front view of a handheld EP probe 100 according to an embodiment of the present invention. The EP probe comprises cylindrical housing 101 and an electrode assembly 102. The housing 101 having a proximal end 101a and a distal end 101b. The electrode assembly 102 is releasably mounted at the proximal end of the cylindrical housing 101.
[0058] The “EP probe” further comprises an electrode assembly releasing and retaining means 103, which is mounted at the distal end 101b of the housing 101. The electrode assembly releasing and retaining means 103 is configured to selectively release the electrode assembly 102 from the housing 101 after use. Such electrode assembly releasing and retaining means 103 preferably include, but not limited to, magnets, push-button, plunger and/or spring, or any other similar releasable fastening means. In an embodiment of the invention, the electrodes assembly 102 is disposable after use with one subject. In an alternative embodiment of the invention, the electrode assembly is adapted to be reused after sterilization by any known method in the art.
[0059] In one embodiment of the present invention, the housing 101 can be molded as one-part housing. In another embodiment of the present invention as shown in FIG. 3a, the housing 101 can be formed as two-part housing, that is formed by releasably fastening two halves together such that a hollow space is created between the two halves.
[0060] The housing 101 further accommodates a pulse generating means 104 which generates a plurality of pulses of energy that are required to be delivered at a target site in a subject. The pulse generating means 104 received power from a power supplying means 105 to generate a plurality of the pulses of the energy.
[0061] In an embodiment of the present invention, the EP probe may further comprise a power supplying means 105 (not shown). In a preferred embodiment of the invention, the power supplying means and the pulse generating means are electrically connected via cable for supplying power to pulse generating means 104. In an alternative embodiment, the power supplying means 105 is adapted to be accommodated within the housing 101. In an embodiment of the invention, the power supplying means 105 can be a battery or PCB.
[0062] The EP probe is further provided with a LED indicator 106 which indicates the mode of operation of the EP probe, especially the mode of operation of the electrode assembly.
[0063] FIG. 3b is a perspective view of the EP probe from the top, front, side and bottom according to an embodiment of the present invention. In a preferred embodiment, the electroporation probe has a cylindrical or elliptical cylinder shape. However, any shape that facilitates easy grip of the probe is within the scope without departing from the spirit of the present invention.
[0064] FIG. 4 and 5 is an isometric and enlarged view of an electrode assembly 102 mounted on the housing 101 in accordance with an embodiment of the present disclosure.
[0065] The electrode assembly 102 comprises a first electrode 102a and a second electrode 102b. The first electrode comprises multiple sub-electrodes 102c which are formed by embossing. The embossed electrodes can be arranged in any geometric array selected from group consisting of a round, a square, an ellipsoid a rectangle, a hexagon, a pentagon and an octagon, more preferably round. The use of embossed electrodes instead of needle electrodes provides increased surface area for delivery of the pulse to enable a non-invasive and painless electroporation in the subject.
[0066] In a preferred embodiment, a total number of sub-electrodes 102c embossed on the first electrode 102a are in a range from 8 to 14, preferably 8 to 10. In a preferred embodiment of the present invention, the first electrodes 102a are configured to deliver the positively charged pulses, while the second electrodes 102b are configured to deliver negatively charged pulses at the target site.
[0067] In a preferred embodiment, the electrode assembly 102 further comprises an electrode connector 102d. The electrode connector has a conical shape, more preferably, a truncated conical shape. That is, electrode connector 102d has first end 102d-i that has first diameter D1 that is greater than the second diameter D2 of the second end 102d-ii. The first end 102d-i of the electrode connector 102d provides means for releasably fastening the electrode assembly 102 to the housing 101. Such fastening means are preferably magnets, hooks, clips (not shown) but can also include other similar releasable fasteners.
[0068] In a preferred embodiment, the second end 102d-ii of the electrode connector is particularly adapted to mount the first electrode 102a and the second electrode 102b in a specific arrangement. The second end 102d-ii of the electrode connector 102d is provided with a central projection (p) and a peripheral ring (r) such that an annular gap (g) is created between the central projection (p) and peripheral ring (r). The first electrode 102a is adapted to be mounted on the central projection (p), while the second electrode 102b is adapted to be mounted on the peripheral ring (r). Here, the annular gap (g) is provided between the central projection (p) and peripheral ring (r) to provide electrical separation between the first electrode 102a and the second electrode 102b.
[0069] This arrangement of the first electrode 102a and the second electrode 102b is crucial for accelerating the electroporation at the target site. The first electrode 102 provided at a center of the probe is configured to deliver positively charged pulses, while the peripherally mounted second electrode 102b is configured to deliver the negatively charged pulse. The annular arrangement of the negative electrode surrounding the positive electrodes generate a plurality of specific paths of current at the target site, that accelerates the generation of desired electric potential at a target site resulting in the formation of larger pores in a short time enabling faster delivery of the biomolecule within the target cells.
[0070] In a preferred embodiment of present invention, the first electrode 102a and the second electrode 102b have a ring-like, annular or tri-podal structure. The first electrode 102a preferably comprises a circular base with a number of electrodes embossed thereon and is adapted to be mounted on the central projection (p) of the electrode connector 102d. Similarly, the second electrode 102b preferably has a ring structure, adapted to be mounted on the peripheral ring (r) of the electrode connector 102d.
[0071] In another preferred embodiment of the present invention, the first electrode 102a and the second electrode 102b are high-frequency switchable electrodes with dual modes of operation. That is, both of the first electrode 102a and the second electrode 102b can operatively switch from the first mode of operation to the second mode of operation. In the first mode, the first electrode 102a and the second electrode 102b are configured to sense an electrical charge present at the target tissue, while in the second mode, the electrodes are configured to deliver dynamic pulses of energy at the target site. In a preferred embodiment, the pulse management system 200 is configured to control switching of the first electrode 102a and the second electrode 102b from the first mode of operation to the second mode of operation depend upon the effective trans-membrane potential needs to generate at the target site.
[0072] In a preferred embodiment of the invention, the first electrode and the second electrode are made up of gold or gold plated material.
[0073] FIG. 6 is an exploded view of an electrode assembly in a dis-assembled state in accordance with an embodiment of the present invention. FIG. 6 particularly shows the arrangement of ring-type second electrode 102b and first electrode 102a at the second end 102d-ii of the electrode connector 102d. In another embodiment of the invention, the electrode connector 102d is made of electro-insulative or non-conductive material.
[0074] To achieve efficient electroporation, the first electrode 102a and the second electrode 102b are provided with a non-parallel placement. As can be seen from FIG. 6, central projection (p) having a height less than the peripheral ring (r). Accordingly, the first electrode 102 will be placed slightly deeper than the second electrode. This is particularly meant to enable the first electrode 102 to encircle the tissue bulge formed at the target site immediately after biomolecule delivery, while the second electrodes are provided at a slight height which facilitates contact with the skin surrounding the bulge. This will particularly enhance the electroporation as the bulge is in contact of the positive charged electrode while encircled by the negatively charged electrode. This will create different paths of current that facilitate creation of stable larger electro-pores in a short period of time while demanding lesser number of pulses of energy. Thus avoids adverse effect to the nearby cells at the target site.
[0075] In an alternative embodiment of the present invention, the first electrode 102a and the second electrode 102b may have a tri-podal structure. That is, both electrodes have a circular or annular base from which three or more legs are extended. While mounting, the base of the electrode rests on the second end of the electrode connector 102d-ii, while the legs thereof extend towards the first end 102d-i and act as an engaging means for the electrodes on the electrode connector 102d.
[0076] Electrode assembly 102 further comprises an LED indicator 106. LED indicator 106 is adapted to indicate the operational mode of the EP probe, more preferably the first electrode 102a and the second electrode 102b, by producing different color light. For example, LED indicator can preferably indicate Red, Green, and Yellow light. Here, the red light indicates the EP probe is in the first mode of operation, green indicates the EP probe is in the second mode of operation and yellow light indicates the completion of the electroporation.
[0077] FIG. 7 is a perspective view of electrode assembly from the front and bottom view in accordance with an embodiment of the present disclosure.
[0078] FIG. 8 flow chart illustrating a method of performing dynamic electroporation for accelerating the delivery of biomolecules into cells of a target site of a subject in accordance with an embodiment of the present invention.
[0079] The method specifically comprises a step of providing subject information to the pulse management system 200 and a step of delivering a predetermined amount of biomolecules in the subject at the target site. Both of the aforesaid steps can be done simultaneously, concurrently or sequentially. The subject information can also be retrieved from storage databases which store the information related to the subject collected in past either manually or by other retrieval software known in the art. The target site is preferably, intra dermal or muscular. After this, handheld EP probe 100 is placed at the target site to perform a method 80 for dynamic electroporation for accelerating the delivery of biomolecules into the cells of target site of a subject. The method 80 comprises following steps which are discussed in detail one-by-one hereinafter.
[0080] Step 801 – sensing a base electrical charge present at the target site.
[0081] As it is known in the art, each of the biomolecules carries more or less charge, preferably a negative charge. Hence, the charge measured at the target site after delivery of the biomolecule represents the amount of active biomolecule delivered at the target site.
[0082] Step 802 - comparing the base electrical charge measured with a first threshold, wherein the first threshold is determined based on the subject information and the first charge information.
[0083] In a preferred embodiment of the present invention, the first threshold is determined by the pulse management system 200 based on the subject information and a first charge information. The first charge information provides a charge of total amount of biomolecules (tB) that is administered at the target site. This step is performed to quantify the total amount of active biomolecule (tB) that has been actually delivered to the subject at a target site.
[0084] If the base charge measured at step 801 is lesser than the first threshold, it indicates that the subject did not receive a required dose of biomolecules and then step of termination 809 of the process will performed with display of an error message. This is particularly advantageous as this step provides the subject with a measure to ensure the solution he is administered with is not a placebo or saline solution and indeed contains a required dose of active biomolecules he intended/paid for. This will significantly forbears deceitful practices, by certain dishonest medical practioners or traders, who trade a saline or placebo solution at a whooping charge of active biomolecules.
[0085] Step 803 – delivering a first set of the plurality of dynamic pulses of energy at the target site, if the base electrical charge is greater than the first threshold;
[0086] If the base charge measured at the target site is greater than the first threshold, it indicates that the target site contains at least the required amount of active biomolecules capable of being entrapped within the target cells. A plurality of pulses of energy is delivered at a target site at step 803, which is sufficient to generate an effective trans-membrane potential at the target site to promote entrapment of all or some of active biomolecules within the target cells. Here, the number of pulses of energy that is needed to be delivered at a target site is determined based on the subject information and the base electrical charge measured at the target site.
[0087] In a preferred embodiment, the pulse management system 200 is configured to determine the first threshold and accordingly schedule the delivery of a plurality of pulses of energy at the target site based on the subject information, the electrical charge of the biomolecules, and the base electrical charge measured at the target site. The pulse management system 200 is configured to dynamically schedule the pulse parameters such as voltage, pulse length, number, and frequency of pulses based on the subject information and the base electrical charge measured at the target site.
[0088] Step 804 - sensing a residual electrical charge present at the target site after delivery of a first set of dynamic pulses of energy;
[0089] The residual electrical charge represents the charge of biomolecules that are not entrapped by the target cells, i.e. biomolecules remaining at the target site after the application of the first set of a plurality of pulses of energy. This step is particularly important for carrying out the invention as it reflects how the target cells of the subject actually reacts to the first set of pulses of the energy delivered at step 803.
[0090] Step 805 - comparing the residual electrical charge measured with a second threshold, wherein the second threshold is determined based on the subject information and a second charge information;
[0091] In a preferred embodiment of the invention, the second threshold is determined by the pulse management system 200 based on the subject information and a second charge information. The second charge information provides the total number of biomolecules that need to be administered inside the cells at the target site.
[0092] Step 806 - delivering a second set of a plurality of dynamic pulses of energy at the target site, if the residual electrical charge measured is greater than the second threshold.
[0093] If the residual electrical charge measured is greater than the second threshold, it indicates that only a portion of biomolecules is entrapped within the target cells, than actually required to be entrapped as provided by the second charge information. In such cases, the process will continue to perform step 806, that is to deliver a second set of pulses of energy is delivered. The second set of pulses of energy is scheduled in a manner that promotes entrapment of higher number of biomolecules than step 803.
[0094] Step 807 - iteratively repeating the above step 803 to step 806 until the residual electrical charge measured at the target site is lesser than the second threshold. Here, the delivery of the plurality of dynamic pulses of energy is scheduled based upon on at least one of subject information, the electrical charge of the biomolecules, and an electrical charge measured at the target site. The pulse parameters can be recorded using an oscilloscope to verify the system performance and to further train the database 202 for future use.
[0095] Step 808 - Process ends when the residual electrical charge measured at the target site is less than the second threshold.
[0096] As aforesaid, the residual charge indicates the number or amount of biomolecules present at the target site, That is, the residual charge less than the second threshold indicates that no biomolecules are present at the target site or required numbers of biomolecules are already delivered during the first set of delivery of pulses of energy, the step of terminating the process (808) is performed and the process is deemed to be completed. The completion of the process is indicated by the LED indicator by emitting yellow light. This is followed by removing the electrode assembly 102 from EP probe 100 by activating the electrode assembly releasing and retaining means 103.
[0097] In a preferred embodiment, the step 801 and 804 are performed by operating the first electrode 102a and the second electrode 102b in the first mode of operation by the pulse management system 200. Similarly, step 803 and 804 are performed by operating the first electrode 102a and the second electrode 102b in the second mode of operation by the pulse management system 200. Steps 802, 805, and 807 are performed by the pulse management system 200.
[0098] The method further comprises step 809 of terminating the process after step 802 by the pulse management system 200, if the base electrical charge measured at the target site is lesser than the first threshold.
[0099] The first charge information provides the charge of the total amount of biomolecules (tB) that is administered at the target site. The pulse management system is configured to determine first charge information based on the number of biomolecules contained in the biomolecule delivery means (such as injection vial) and the charge carried by a single biomolecule.
[00100] The second charge information provides the charge of total amount of biomolecules (tB) that needs to be entrapped within the cells at of the target site. The pulse management system is configured to determine the second charge information based on subject information and the first charge information.
[00101] In simpler words, let’s assume that,
tB is total amount of biomolecules that are present in a delivery means.
dB is an amount of biomolecules that needs to be delivered as suggested by subject information and electrical charge measured at the target site.
pB is an amount of biomolecules actually delivered i.e., present in an active state at the target site.
rB is an amount of biomolecules remnant after the dB is administered in the target cells of the target site.
Here, tB=dB+pB+rB
[00102] Further, pB is determined by the pulse management system 200 by operating the first electrode 102a and the second electrode 102b in the first mode of operation. dB is calculated by the pulse management system 200 based on the subject information and pB, i.e., the amount of biomolecules actually delivered i.e., present in an active state at the target site.
[00103] Here, three situations are possible
i) pB=dB, then pulse management system 200 continue to deliver the pulses of energy until all number of pB is entrapped within the target cells at the target site;
ii) pB>dB, then pulse management system 200 will continue to deliver the pulses of energy until dB number of biomolecule out of pB is entrapped within the target cells at the target site. Remaining biomolecules are designated at rB and may or may not require to deliver inside the target cells to develop the required biological effect;
iii) pB
Documents
Orders
Section
Controller
Decision Date
Application Documents
#
Name
Date
1
202121040857-OTHERS [17-09-2024(online)].pdf
2024-09-17
1
202121040857-PROOF OF RIGHT [09-09-2021(online)].pdf
2021-09-09
2
202121040857-FORM FOR SMALL ENTITY [04-09-2024(online)].pdf
2024-09-04
2
202121040857-POWER OF AUTHORITY [09-09-2021(online)].pdf
2021-09-09
3
202121040857-IntimationOfGrant16-05-2024.pdf
2024-05-16
3
202121040857-FORM FOR SMALL ENTITY(FORM-28) [09-09-2021(online)].pdf
2021-09-09
4
202121040857-PatentCertificate16-05-2024.pdf
2024-05-16
4
202121040857-FORM FOR SMALL ENTITY [09-09-2021(online)].pdf
2021-09-09
5
202121040857-Written submissions and relevant documents [11-03-2024(online)].pdf
2024-03-11
5
202121040857-FORM 1 [09-09-2021(online)].pdf
2021-09-09
6
202121040857-EVIDENCE FOR REGISTRATION UNDER SSI(FORM-28) [09-09-2021(online)].pdf
2021-09-09
6
202121040857-Correspondence to notify the Controller [20-02-2024(online)].pdf