CN108290036B

CN108290036B - Electroporation apparatus and control method thereof

Info

Publication number: CN108290036B
Application number: CN201680067088.6A
Authority: CN
Inventors: 李承俊
Original assignee: Sl Vaxigen Inc
Current assignee: SL VAXIGEN, Inc.
Priority date: 2015-09-17
Filing date: 2016-09-13
Publication date: 2022-02-18
Anticipated expiration: 2036-09-13
Also published as: KR20170033703A; CN108290036A; WO2017048037A1; KR101788301B1

Abstract

The present invention relates to an electroporation apparatus and a control method thereof. The electroporation apparatus includes: a penetrating unit including a plurality of electrodes for penetrating into a living body; and a controller for controlling a voltage to be applied to each of the plurality of electrodes, and applying a pulse by setting one of the plurality of electrodes to have a reference polarity and the other electrodes to have a polarity opposite to the reference polarity.

Description

Electroporation apparatus and control method thereof

Technical Field

The present invention relates to an electroporation apparatus and a control method thereof, and more particularly to an electroporation apparatus for delivering an implant agent into a tissue or a cell using an electric pulse and a control method thereof.

Background

Electroporation is a technique employed in the field of molecular biology whereby an electric field is applied to a cell to increase the transmissivity of the cell membrane so that chemically, medically, biologically active molecules, etc. may be introduced into the cell.

Electroporation can be used for gene delivery, DNA vaccine vaccination, etc., similar to iontophoresis therapy, lipid delivery, or gene gun (or particle gun). That is, when a short and strong electric pulse is applied to a cell, hydrophilic pores are generated at a lipid bilayer of the cell, and thus even charged molecules or macromolecules such as nucleic acids can be delivered to the cell.

The background art of the present invention is disclosed in korean patent laid-open publication No. 10-2014-0048213 (published: 2014 4/23).

Disclosure of Invention

[ problem ] to provide a method for producing a semiconductor device

The electroporation described above may be produced by applying a relatively strong electric field. In particular, since the difference between the threshold intensity of the pulse for generating the hydrophilic pores and the intensity of the pulse for destroying the cells is small, the pulse as low as possible should be used in the range of generating the hydrophilic pores when the drug is delivered to the living organism using the electroporation for the sake of safety.

That is, in this case, hydrophilic pores may be generated only in the vicinity of the line connecting the positive and negative electrodes of the electroporation apparatus, and hydrophilic pores are not generated in the region spaced a certain distance or more from the line because an electric field having less than a threshold intensity is applied to the region.

However, in general, a common electroporation apparatus includes a pair of positive and negative electrodes, and applies an electric field of a fixed form. Thus, the region in which the hydrophilic pores are generated is limited.

To solve this problem, a method using a plurality of electrodes having a plurality of pairs of positive and negative electrodes is generally employed. However, interference is likely to occur between a plurality of electrodes, and thus it is limited to cumulatively arrange the electrodes in the same area. Accordingly, it is difficult to improve the ability to deliver drugs to the body without increasing the area of the electrode penetrating the body.

To solve the above-described problems of the conventional electroporation apparatus, the present invention is directed to an electroporation apparatus and a control method thereof capable of improving the ability to deliver a drug without increasing the number of electrodes.

[ technical solution ] A

One aspect of the present invention provides an electroporation apparatus comprising: a penetrating unit including a plurality of electrodes for penetrating into a living body; and a controller for controlling a voltage to be applied to each of the plurality of electrodes, and applying a pulse by setting one of the plurality of electrodes to have a reference polarity and the other electrodes to have a polarity opposite to the reference polarity.

In the present invention, when the electrode having the reference polarity is a positive electrode, the electrode having the opposite polarity may be a negative electrode, and when the electrode having the reference polarity is a negative electrode, the electrode having the opposite polarity may be a positive electrode.

In the present invention, the plurality of electrodes may be arranged in the same form in the relative relationship of the distances therebetween, regardless of the reference electrode.

In the present invention, the plurality of electrodes may be arranged in a rectangular shape.

In the present invention, the penetration unit may further comprise an injection needle for injecting the introducer.

In the present invention, each of the plurality of electrodes may be in the form of an injection needle for injecting an introducer.

In the present invention, the introducing agent may be a biologically active molecule and may comprise a peptide or a nucleic acid, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), and is preferably a DNA vaccine, messenger ribonucleic acid (mRNA) or antisense RNA (microribonucleic acid microRNA or small interfering ribonucleic acid siRNA).

In the present invention, the plurality of electrodes may be used to penetrate the skin (intradermally), muscle (intramuscularly) or tumor (intratumorally).

In the present invention, the controller can sequentially change the electrodes to have a reference polarity.

In the present invention, the controller may change the electrodes to have the reference polarity in a clockwise direction or a counterclockwise direction.

In the present invention, the controller may change the electrodes to have the reference polarity after a pause period during which no pulse is applied.

In the present invention, the controller may apply pulses, the number of pulses being equal to the number of other electrodes, and the pulses may be applied using different electrodes in units of pulses in the other electrodes.

In the present invention, the controller may sequentially change the electrodes to which the pulses are to be applied in a clockwise direction or a counterclockwise direction among the other electrodes.

In the present invention, the controller may have a time interval between pulses during which no pulse is applied.

Another aspect of the present invention provides a method of controlling an electroporation device, comprising: a first operation of applying a pulse by setting one of a plurality of electrodes to have a reference polarity and the other electrodes to have a polarity opposite to the reference polarity; a second operation of applying a pulse by setting one of the other electrodes having an opposite polarity in the first operation to have a reference polarity and setting the other electrodes to have a polarity opposite to the reference polarity; and a third operation of applying a pulse by setting one of the electrodes having opposite polarities of both the first and second operations to have a reference polarity and setting the other electrode to have a polarity opposite to the reference polarity, the first to third operations being performed by the controller.

In the present invention, the method may further comprise pausing, by the controller, for one pause period after the first operation and before the second operation, not applying the pulse; and pausing, by the controller, the pause period after the second operation and before the third operation without applying the pulse.

In the present invention, the first operation may include: operation S1 — applying a pulse through one of the other electrodes and the electrode having the reference polarity; and operation S2 — applying a pulse through one of the other electrodes that is not used in operation S1 and the electrode having the reference polarity, operations S1 and S2 being performed by the controller.

In the present invention, before the first operation, the method may further include penetrating the plurality of electrodes and the injection needle into skin (intradermally), muscle (intramuscularly), or tumor (intratumorally) by a controller, and injecting an implant through the plurality of electrodes and the injection needle by the controller.

[ PROBLEMS ] the present invention

In the electroporation apparatus and the control method thereof according to the present invention, pulses are applied by setting one of the plurality of electrodes to have a reference polarity and the other electrodes to have a polarity opposite to the reference polarity, the electrodes to have the reference polarity are sequentially changed, and different electrodes are used in the form of several pulse units, thereby improving the ability of the electroporation apparatus to deliver an implant agent.

Drawings

Fig. 1 is a block diagram of the structure of an electroporation apparatus according to an embodiment of the present invention.

Fig. 2A is a schematic view of a penetration unit of an electroporation device according to an embodiment of the present invention.

Fig. 2B is a schematic view of a penetration unit of an electroporation apparatus according to another embodiment of the present invention.

Fig. 3 is a schematic diagram of a pulse application method performed by an electroporation apparatus according to an embodiment of the present invention.

Fig. 4 is another schematic diagram of a pulse application method performed by an electroporation apparatus according to an embodiment of the invention.

Figure 5 is a flow diagram of a method of controlling an electroporation device according to one embodiment of the invention.

Fig. 6 is a flowchart of a pulse application method employed in a method of controlling an electroporation apparatus according to an embodiment of the present invention.

Fig. 7 is another flowchart of a pulse application method employed in a method of controlling an electroporation apparatus according to an embodiment of the present invention.

Detailed Description

Hereinafter, an electroporation apparatus and a control method thereof according to an embodiment of the present invention will be described with reference to the accompanying drawings. Here, the thickness of each line or the size of each component illustrated in the drawings may be exaggerated for clarity and convenience of explanation. The terms used herein are defined in consideration of functions of the present invention, but may be defined differently according to the intention of a user or operator or according to precedent. Accordingly, these terms should be defined based on the entire context of the present invention.

Fig. 1 is a block diagram of the structure of an electroporation apparatus according to an embodiment of the present invention. Fig. 2A is a schematic view of a penetration unit of an electroporation device according to an embodiment of the present invention. Fig. 2B is a schematic view of a penetration unit of an electroporation apparatus according to another embodiment of the present invention. Fig. 3 is a schematic diagram of a pulse application method performed by an electroporation apparatus according to an embodiment of the present invention. Fig. 4 is another schematic diagram of a pulse application method performed by an electroporation apparatus according to an embodiment of the invention. The electroporation apparatus according to the present embodiment will be described below with reference to the drawings.

First, as shown in fig. 1, an electroporation apparatus according to an embodiment of the present invention includes a controller 100 and a penetration unit 110. The penetration unit 110 may include first to nth electrodes 111 to 118, and an injection needle 119.

The

electrodes

111, 112, 113, 114, and 118 of the penetrating unit 110 and the injection needle 119 may penetrate into the living body under the control of the controller 100. For example, the

electrodes

111, 112, 113, 114 and 118 and the injection needle 119 may penetrate the skin by being sucked into the skin (intradermal penetration) by a suction assembly (not shown) driven under the control of the controller 100, may penetrate the muscle where it is moved (intramuscular penetration) by a motor assembly (not shown), or may penetrate the tumor (intratumoral penetration).

The introduction agent may be injected into a living body through an injection needle 119 under the control of the controller 100. For example, the injection needle 119 may be in the form of a common injection needle. That is, the electroporation apparatus according to the present embodiment may include a syringe (not shown) containing the implant agent, and may thus inject the implant agent into the living body.

Alternatively, as shown in fig. 2A and 2B, a plurality of injection needles 119 may be provided, which may preferably be located in the region where the

electrodes

111, 112, 113 and 114 are connected. As described above, as the distance between the region and the electrode becomes shorter, hydrophilic pores are more likely to be generated. Therefore, when the implant agent is present in this region, the implant agent can be smoothly delivered into the cell.

Here, the implant agent is an agent to be delivered to a target tissue or cell by electroporation. The implant agent can be a substance (e.g., a chemical, cosmetic, bioactive molecule, etc.) that produces an activity or function when introduced into a tissue or cell.

A biologically active molecule is a molecule that produces a biological effect when introduced into a cell and may include, but is not limited to, a nucleic acid of DNA or RNA, an antigenic peptide, an antibody, or an antibody fragment. Additional examples of biologically active molecules include nucleic acids, such as plastids, encoding nucleic acid sequences, mRNA, and antisense RNA molecules.

Nucleic acids refer not only to analogs of RNA or DNA generated from nucleotide analogs in single-or double-stranded form, but also to oligonucleotides or polynucleotides, such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), and include DNA, mRNA, antisense RNA (mirrna or siRNA).

The

electrodes

111, 112, 113, 114, and 118 may be penetrated into a living body under the control of the controller 100 and apply an electric pulse into the living body to deliver an implant agent into a target tissue or cell.

The polarity of the

electrodes

111, 112, 113, 114 and 118 may not be fixed. That is, the controller 100 may control a voltage to be applied to each of the

electrodes

111, 112, 113, 114, and 118, thereby setting the polarity of each of the electrodes to positive or negative.

The

electrodes

111, 112, 113, 114 and 118 may be fabricated in a form in which the implant can be injected, similar to the injection needle 119. For example, each of the

electrodes

111, 112, 113, 114, and 118 may be in the form of a common injection needle, similar to injection needle 119. As described above, as the distance between the implant agent and the electrode becomes shorter, the likelihood of delivery of the implant agent into the cell increases. Therefore, when the implant agent is injected through the

electrodes

111, 112, 113, 114, and 118, the efficiency of electroporation can be improved. In another embodiment of the present invention, the penetration unit 110 may not additionally include the injection needle 119.

The penetration unit 110 may be in a form that can be separated from the electroporation apparatus according to the present embodiment. That is, the penetration unit 110 is used to penetrate into a living body, and thus may be designed in the form of a disposable product to prevent infection.

When performing electroporation, the controller 100 may apply pulses by setting one of the

electrodes

111, 112, 113, 114, and 118 to have a reference polarity and the other electrodes to have a polarity opposite to the reference polarity. That is, when the electrode having the reference polarity is a positive electrode, the electrode having the opposite polarity is a negative electrode, and when the electrode having the reference polarity is a negative electrode, the electrode having the opposite polarity is a positive electrode.

In other words, the controller 100 may perform electroporation by setting one of the plurality of electrodes as a positive electrode and the other electrodes as negative electrodes or by setting one of the plurality of electrodes as a negative electrode and the other electrodes as a positive electrode. When only one reference polarity electrode is used as described above, an electric field can be formed in one direction, and the possibility of interference between the electric fields can be lower than when a plurality of positive electrodes and a plurality of negative electrodes are used.

The case where the reference polarity electrode is a positive electrode will be described below. The description will also apply to the case where the reference polarity electrode is a negative electrode. Although the number of electrodes is not limited in the present invention, a case where four

electrodes

111, 112, 113, and 114 are provided will be described as an example for convenience of explanation.

That is, the controller 100 may perform electroporation by setting only the first electrode 111 as a positive electrode and the second to

fourth electrodes

112, 113 and 114 as negative electrodes or by setting only the third electrode 113 as a positive electrode and the first, second and

fourth electrodes

111, 112 and 114 as negative electrodes.

Alternatively, the controller 100 may sequentially change the electrodes to have a reference polarity for electroporation. That is, as shown in fig. 3, the controller 100 may apply the pulse by performing one set of setting the first electrode 111 as a positive electrode and the second to

fourth electrodes

112, 113 and 114 as negative electrodes, and by performing another set of setting the second electrode 112 as a positive electrode and the first, third and

fourth electrodes

111, 113 and 114 as negative electrodes. That is, the controller 100 may perform electroporation by performing the sets while changing the electrodes to have the reference polarity.

In this case, the controller 100 may sequentially change the electrodes to have the reference polarity in a counterclockwise direction as shown in fig. 3 or in a clockwise direction.

Further, when one group is completed, the controller 100 may perform another group after a pause period during which no pulse is applied. That is, the cells may disappear due to an increase in temperature when the electric field is continuously applied. Thus, for safety, the controller 100 may set a pause period from group to group. Typically, this pause period may be set to be longer than the duration of the group. For example, the duration of one group may be designed to be 30ms, and the pause period may be designed to be 100 ms.

As described above, a plurality of lines connecting the positive and negative electrodes may be formed by changing the electrodes to have a reference polarity, thereby creating hydrophilic pores in wider regions. Thus, the ability of the electroporation device to deliver an implant agent may be improved when using this method. Furthermore, even if a relatively low electric field is applied (even if the area of each line in which hydrophilic pores can be generated is small), all the areas in which hydrophilic pores can be generated can be maintained at the same level as in the conventional method, so that electroporation can be performed more safely.

In order to apply an electric field of the same strength regardless of the positions of the electrodes, a voltage proportional to the distance between the positive and negative electrodes should be applied to a pair of electrodes. That is, if the relationship of the distance between the positive electrode and the negative electrode changes each time the position of the positive electrode changes in rotation, an electric field of the same strength can be formed only when a different voltage is applied for each of the groups. In this case, the user will feel discomfort, and the structure of the circuit controlling the voltage to be applied to each electrode may be complicated. Accordingly, in the present embodiment, the plurality of electrodes may be arranged in the same form of the relative relationship of the distances between the electrodes, regardless of the electrode as the reference electrode.

That is, the relative relationship of the distances between the electrodes is the same regardless of the electrode as the reference electrode, which is understood to mean that when each of the electrodes is set as the reference electrode, the types of distances to the other electrodes are the same. For example, in the case where the

electrodes

111, 112, 113, and 114 are arranged in a rectangular shape as shown in fig. 3, when the first electrode 111 is a reference electrode, the types of distances to the other electrodes are three: horizontal, vertical and diagonal lines. Similarly, when the second electrode 112 is a reference electrode, there are only three types of distances to other electrodes: horizontal, vertical and diagonal lines. That is, when the

electrodes

111, 112, 113, and 114 are arranged in a rectangular shape, the controller 100 may maintain the overall strength of the electric field constant using only voltages corresponding to the three types of distances.

When one set is performed, the controller 100 may apply pulses at the same time using different electrodes in the form of several pulse units among other electrodes, the number of pulses being equal to the number of other electrodes having a polarity opposite to that of the reference electrode. That is, in fig. 4, the electrodes indicated by dotted lines represent the electrodes to which no pulse is applied. As shown in fig. 4, the controller 100 may apply three pulses to perform the first group such that the pulses are applied through the first and

second electrodes

111 and 112, the pulses are applied through the first and

third electrodes

111 and 113, and the pulses are applied through the first and

fourth electrodes

111 and 114.

In this case, the controller 100 may sequentially change the electrodes to which the pulses are to be applied in a counterclockwise direction as shown in fig. 4, or may apply the pulses in a clockwise direction.

Further, the controller 100 may set a time interval between pulses during which no pulses are applied. That is, because the shape of the pulse is more similar to the shape of a Direct Current (DC), the cells are more likely to be damaged. Therefore, for safety, the controller 100 may control the application of the next pulse after a time interval after the application of one pulse. In this case, the time interval may more closely approximate the duration (e.g., 10ms) in which one pulse is applied than the scale of the pause period (pause duration).

When one negative electrode is used to apply one pulse as described above, the influence caused by interference between the negative electrodes can be reduced, and thus the electrodes can be arranged cumulatively in a narrow region.

Fig. 5 is a flow chart of a method of controlling an electroporation device according to an embodiment of the invention. Fig. 6 is a flowchart of a pulse application method employed in a method of controlling an electroporation apparatus according to an embodiment of the present invention. Fig. 7 is another flowchart of a pulse application method employed in a method of controlling an electroporation apparatus according to an embodiment of the present invention. A method of controlling an electroporation apparatus according to the present embodiment will be described below with reference to the drawings.

As shown in fig. 5, first, the controller 100 causes the

electrodes

111, 112, 113, and 114 and the injection needle 119 to penetrate into the skin, muscle, or tumor (operation S500). For example, the controller 100 may penetrate the

electrodes

111, 112, 113, and 114 and the injection needle 119 into the skin (intradermal penetration) by sucking the skin using a suction assembly (not shown), may penetrate the

electrodes

111, 112, 113, and 114 and the injection needle 119 into the muscle (intramuscular penetration) by moving the

electrodes

111, 112, 113, and 114 and the injection needle 119 using a motor assembly (not shown), and may penetrate the

electrodes

111, 112, 113, and 114 and the injection needle 119 into the tumor (intratumoral penetration).

Next, the controller 100 injects an implant through the

electrodes

111, 112, 113, and 114 and the injection needle 119 (operation S510). Each of the

electrodes

111, 112, 113 and 114 may be in the form of an injection needle, similar to injection needle 119. Here, the implant agent refers to an agent to be delivered to a target tissue or cell by electroporation, and may be a chemical, a DNA vaccine, or the like.

After operation S510, the controller 100 introduces an implant agent into the target tissue or cell by applying pulses through the

electrodes

111, 112, 113, and 114 (operation S520). That is, the controller 100 may perform electroporation by applying pulses through the

electrodes

111, 112, 113, and 114. The above-described operation S520 will be described in more detail with reference to fig. 6.

As shown in fig. 6, first, the controller 100 applies a pulse by setting the first electrode 111 as a positive electrode and the

other electrodes

112, 113, and 114 as negative electrodes (operation S600). That is, the controller 100 may apply the pulse by setting one of the

electrodes

111, 112, 113, and 114 (e.g., the first electrode 111) to have a reference polarity and setting the

other electrodes

112, 113, and 114 to have a polarity opposite to the reference polarity. The above-described operation S600 will be described in more detail below with reference to fig. 7.

As shown in fig. 7, first, the controller 100 applies a pulse through the first electrode 111 and the second electrode 112 (operation S700). That is, the controller 100 may apply the pulse by setting one of the

electrodes

112, 113, and 114 having opposite polarities (e.g., the second electrode 112) through operation S600 of fig. 6 and the electrode (the first electrode 111) set as the electrode having the reference polarity in operation S600 of fig. 6.

Next, the controller 100 applies a pulse through the first electrode 111 and the third electrode 113 (operation S710). That is, the controller 100 may set one of the electrodes 113 and 114 (e.g., the third electrode 113) that is not used in operation S700 among the electrodes having opposite polarities and the electrode set to the electrode having the reference polarity in operation S600 of fig. 6 to apply the pulse through operation S600 of fig. 6.

After operation S710, the controller 100 applies a pulse through the first electrode 111 and the fourth electrode 114 (operation S720). That is, the controller 100 may perform operation S600 of fig. 6 by applying pulses while sequentially changing electrodes to which the pulses are applied in a counterclockwise direction, the number of pulses being equal to the number of electrodes set as negative electrodes.

Further, the controller 100 may set a time interval between operations S700 and S710 and between operations S710 and S720 during which no pulse is applied.

After operation S600 of fig. 6, the controller 100 has a pause period (operation S610). That is, when the electric field is continuously applied to the cells, the cells may disappear due to an increase in temperature, and thus, the controller 100 may have a pause period between groups during which no pulse is applied for safety.

Next, the controller 100 applies a pulse by setting the second electrode 112 as a positive electrode and the

other electrodes

111, 113, and 114 as negative electrodes (operation S620). That is, the controller 100 may apply the pulse by setting one of the

electrodes

112, 113, and 114 (the second electrode 112) set to have an opposite polarity in operation S600 to have a reference polarity and setting the

other electrodes

111, 113, and 114 to have an opposite polarity to the reference polarity.

After operation S620, the controller 100 has a pause period again (operation S630), and applies a pulse by setting the third electrode 113 as a positive electrode and the

other electrodes

111, 112, and 114 as negative electrodes (operation S640). That is, the controller 100 may apply the pulse by setting one of the electrodes 113 and 114 (the third electrode 113) set to have opposite polarities in both operations S600 and S620 to have the reference polarity and setting the

other electrodes

111, 112, and 114 to have a polarity opposite to the reference polarity.

Then, the controller 100 has a pause period again (operation S650), and applies a pulse by setting the fourth electrode 114 as a positive electrode and the

other electrodes

111, 112, and 113 as negative electrodes (operation S660). That is, the controller 100 may perform electroporation by sequentially changing the electrodes to have the reference polarity, and complete electroporation after all the electrodes serve as the electrodes having the reference polarity.

As described above, in the electroporation apparatus and the control method thereof according to the embodiments of the present invention, pulses are applied by setting one of the plurality of electrodes to have a reference polarity and the other electrodes to have a polarity opposite to the reference polarity while sequentially changing the electrodes to have the reference polarity, and different electrodes are used in units of the pulses, thereby reducing the possibility of interference between the electrodes and increasing the region in which hydrophilic pores are generated. Accordingly, the capabilities of the electroporation device for delivering the implant agent may be improved.

While the invention has been described above with reference to the embodiments shown in the drawings, the embodiments are merely examples, and it will be apparent to one of ordinary skill in the art that various modifications and equivalent embodiments can be made. Accordingly, the technical scope of the present invention should be defined in the appended claims.

Claims

1. An electroporation device for delivering an implant agent into a tissue or cell by applying electrical pulses through electrodes, the electroporation device comprising:

a penetrating unit including a plurality of electrodes for penetrating into a living body; and

a controller for controlling a voltage to be applied to each of the plurality of electrodes and applying a pulse by setting one of the plurality of electrodes to have a reference polarity and setting the other electrodes to have a polarity opposite to the reference polarity;

wherein the controller is further arranged to apply the pulses in a manner such that: the number of pulses applied by the controller is equal to the number of the other electrodes; and applying the pulse using different electrodes in units of several pulses among the other electrodes.

2. An electroporation device as claimed in claim 1, wherein the electrode of the opposite polarity is a negative electrode when the electrode of the reference polarity is a positive electrode and a positive electrode when the electrode of the reference polarity is a negative electrode.

3. An electroporation device as claimed in claim 1, wherein the plurality of electrodes are arranged in the same relative relationship with respect to the distance therebetween, irrespective of the reference electrode.

4. An electroporation device as claimed in claim 3, wherein the plurality of electrodes are arranged in a rectangular shape.

5. An electroporation device according to claim 1, wherein the penetration unit further comprises an injection needle for injecting the introducer.

6. An electroporation device as claimed in claim 1, wherein each of the plurality of electrodes is in the form of an injection needle for injecting the introducer.

7. An electroporation device as claimed in claim 1, wherein the introduction agent comprises a DNA vaccine, mRNA or antisense RNA,

wherein the antisense RNA comprises microRNA or siRNA.

8. An electroporation device as claimed in claim 1, wherein the plurality of electrodes are for penetrating skin, muscle or a tumour.

9. An electroporation apparatus as claimed in claim 1, wherein the controller sequentially changes electrodes to have a reference polarity.

10. An electroporation device as claimed in claim 9, wherein the controller changes the electrodes to have a reference polarity in a clockwise or counterclockwise direction.

11. An electroporation device as claimed in claim 9, wherein the controller changes the electrodes to have a reference polarity after a pause period during which no pulses are applied.

12. An electroporation apparatus as claimed in claim 1, wherein the controller sequentially changes electrodes to which the pulses are to be applied in a clockwise direction or a counterclockwise direction among the other electrodes.

13. An electroporation device as claimed in claim 1, wherein the controller has a time interval between the pulses, wherein no pulses are applied during the time interval.