WO2012056756A1 - 導入対象物質の送達装置の作動方法および導入対象物質の送達方法 - Google Patents
導入対象物質の送達装置の作動方法および導入対象物質の送達方法 Download PDFInfo
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- WO2012056756A1 WO2012056756A1 PCT/JP2011/062102 JP2011062102W WO2012056756A1 WO 2012056756 A1 WO2012056756 A1 WO 2012056756A1 JP 2011062102 W JP2011062102 W JP 2011062102W WO 2012056756 A1 WO2012056756 A1 WO 2012056756A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M5/00—Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests
- A61M5/14—Infusion devices, e.g. infusing by gravity; Blood infusion; Accessories therefor
- A61M5/142—Pressure infusion, e.g. using pumps
- A61M5/145—Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. pressurised by means of pistons
- A61M5/14586—Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. pressurised by means of pistons pressurised by means of a flexible diaphragm
- A61M5/14593—Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. pressurised by means of pistons pressurised by means of a flexible diaphragm the diaphragm being actuated by fluid pressure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M5/00—Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M5/00—Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests
- A61M5/14—Infusion devices, e.g. infusing by gravity; Blood infusion; Accessories therefor
- A61M5/142—Pressure infusion, e.g. using pumps
- A61M5/145—Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. pressurised by means of pistons
- A61M5/148—Pressure infusion, e.g. using pumps using pressurised reservoirs, e.g. pressurised by means of pistons flexible, e.g. independent bags
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M60/00—Blood pumps; Devices for mechanical circulatory actuation; Balloon pumps for circulatory assistance
- A61M60/10—Location thereof with respect to the patient's body
- A61M60/122—Implantable pumps or pumping devices, i.e. the blood being pumped inside the patient's body
- A61M60/126—Implantable pumps or pumping devices, i.e. the blood being pumped inside the patient's body implantable via, into, inside, in line, branching on, or around a blood vessel
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M60/00—Blood pumps; Devices for mechanical circulatory actuation; Balloon pumps for circulatory assistance
- A61M60/20—Type thereof
- A61M60/289—Devices for mechanical circulatory actuation assisting the residual heart function by means mechanically acting upon the patient's native heart or blood vessel structure, e.g. direct cardiac compression [DCC] devices
Definitions
- the present invention relates to a method for operating a delivery device for a substance to be introduced and a method for delivering a substance to be introduced.
- nucleic acid medicament a medicament containing the nucleic acid
- nucleic acid medicine is expected to be applied to the treatment of various diseases.
- a method for delivering a nucleic acid to a target site such as an organ for example, a method of directly injecting a nucleic acid into a target site with a syringe (hereinafter referred to as “direct injection method”), a body fluid that is placed in the vasculature of an organ
- direct injection method a method of injecting nucleic acid into a region isolated by blocking the flow
- the direct injection method has a drawback in that the needle around the target site may be damaged by the syringe needle.
- the nucleic acid can be delivered to a region isolated by a catheter, but the nucleic acid is locally delivered only to a target site in an organ or the like. Or it is difficult to deliver nucleic acids to a plurality of target sites at once.
- the present invention has been made in view of the above-described prior art, and can efficiently and locally deliver a substance to be introduced to a target site.
- a method for operating a delivery apparatus for a substance to be introduced and delivery of the substance to be introduced It aims to provide a method.
- the gist of the present invention is as follows.
- An introduction target substance provided with a supply unit that supplies an introduction target substance to a duct located upstream of a target site of a tissue existing in a living body, and a pressure load unit that applies pressure to the target site of the tissue A method of operating a delivery device of (1-1) a step of sending an introduction target substance from a supply unit and supplying the introduction target substance to the pipeline; and (1-2) the same as the step (1-1) or the step (1-1) ), And a method of operating the delivery target substance delivery device including the step of generating a pressure against the target site of the tissue in the pressure load portion, [2]
- the method according to [1], wherein the substance to be introduced is a nucleic acid or a peptide compound
- the pressure load unit includes a suction unit for sucking the target site, In the step (1-2), the method according to the above [1] or [2], wherein the suction site is caused to suck the target site to generate a
- a method for delivering a substance to be introduced that delivers the substance to be introduced to a target site of a tissue of a non-human mammal, (2-1) supplying a substance to be introduced to a conduit located upstream of the tissue, and (2-2) simultaneously with the step (2-1) or after the step (2-1), A method of delivering a substance to be introduced, including a step of applying pressure to a target site of tissue and delivering the substance to be introduced to the target site; [7] The method according to [6], wherein the introduction target substance is a nucleic acid, [8] The method according to [6] or [7], wherein a suction pressure is applied to the target site in the step (2-2), [9] The method according to [6] or [7], wherein the target site is pressed while fixing the position of the tissue in the step (2-2), [10] The tissue is an organ tissue, In the step (2-2), the method according to [9], wherein the target site is pressed while the organ is housed in a case capable of housing
- a method for delivering a substance to be introduced that delivers the substance to be introduced to a target site of a target derived from a living body, (3-1) supplying the substance to be introduced to a site different from the target site in the target, and (3-2) simultaneously with the step (3-1) or after the step (3-1) , Applying a pressure to the target site of the object and delivering the substance to be introduced to the target site;
- a method for delivering a substance to be introduced to a target site of an object comprising: [12] The method according to [11], wherein the introduction target substance is a nucleic acid or a peptide compound, [13] The method according to [11] or [12], wherein a suction pressure is applied to the target site in the step (3-2), and [14] the target site in the step (3-2). The method according to [11] or [12], wherein the pressing is performed while fixing the position.
- the target target substance can be efficiently delivered locally to the target site.
- Example 1 it is a drawing substitute photograph which shows the result of having investigated the expression site
- Example 2 it is a drawing substitute photograph which shows the result of having investigated the expression site
- Example 3 it is a drawing substitute photograph which shows the result of having investigated the expression site
- FIG. 5 is a drawing-substituting photograph showing the results of examining the expression site of luciferase encoded by nucleic acid injected under various conditions in Test Example 1.
- Example 4 it is a graph which shows the result of having investigated the relationship between the kind of used device and luciferase activity.
- A is a schematic explanatory drawing which shows the position in the liver of the part which installed the suction part in Test Example 2.
- FIG. (B) is an enlarged explanatory view of the installation part of the suction part shown in FIG.
- C is a schematic explanatory view showing a section of a portion to be measured for luciferase activity in Test Example 2.
- FIG. In Test Example 2 it is a graph showing the results of examining the relationship between the liver site and luciferase relative activity.
- Experiment 3 it is a graph which shows the result of having investigated the relationship between the kind of inject
- Test example 4 it is a schematic explanatory drawing which shows the structure of the press part used for the load of a pressure.
- Experiment 5 it is a graph which shows the result of having investigated the relationship between pressure load conditions and luciferase activity.
- Example 5 it is a graph which shows the result of having investigated the relationship between the kind of target object, and luciferase activity.
- Example 6 it is a graph which shows the result of having investigated the relationship between the kind of used device and luciferase activity.
- Experiment 6 it is a graph which shows the result of having investigated the relationship between the kind of introduction method, and luciferase relative activity.
- the present inventors locally deliver nucleic acid to a target site by injecting the nucleic acid into a site different from the target site and applying pressure (suction pressure or pressing force) to the target site. Furthermore, the present inventors have found that the protein encoded by the nucleic acid and the function of the nucleic acid can be expressed at the target site.
- the present invention has been made based on such knowledge.
- tissue refers to a tissue that constitutes an organ of an animal having a duct that transports bodily fluids and metabolites, and refers to epithelial tissue, connective tissue, muscle tissue, nerve tissue, and the like.
- tissue refers to epithelial tissue, connective tissue, muscle tissue, nerve tissue, and the like.
- tissue refers to epithelial tissue, connective tissue, muscle tissue, nerve tissue, and the like.
- tissue refers to epithelial tissue, connective tissue, muscle tissue, nerve tissue, and the like.
- tissue refers to epithelial tissue, connective tissue, muscle tissue, nerve tissue, and the like.
- tissue refers to epithelial tissue, connective tissue, muscle tissue, nerve tissue, and the like.
- a pipe refers to animal blood vessels, urinary tracts, lymphatic vessels, and the like.
- a pipe located upstream of a target site refers to a pipe that transports bodily fluids and metabolites to the target site.
- Upstream means the starting point in
- the substance to be introduced is not particularly limited, and examples thereof include nucleic acids, drugs, proteins, peptides or derivatives thereof (hereinafter also referred to as “peptide derivatives”), antibodies, polymer compounds, metals (for example, metal nanomaterials). Particles, etc.), semiconductors (for example, quantum dots, etc.), or substances formed by combining them.
- the “peptide derivative” includes a compound having a peptide skeleton.
- proteins, peptides, and peptide derivatives are collectively referred to as “peptide compounds”.
- nucleic acids or peptide compounds are preferred because they can be efficiently introduced into cells of tissues and organs according to the present invention, although they are usually difficult to permeate cell membranes.
- the present invention provides a supply unit that supplies an introduction target substance to a duct located upstream of a target site of a tissue existing in a living body, and a target site of the tissue.
- a method for operating a delivery device for a substance to be introduced comprising a pressure load section for applying pressure to (1-1) a step of sending an introduction target substance from a supply unit and supplying the introduction target substance to the pipeline; and (1-2) the same as the step (1-1) or the step (1-1) ),
- a method for operating the delivery target substance delivery device including the step of generating a pressure on the target site of the tissue in the pressure load section (hereinafter referred to as “method 1”).
- the method 1 has one major feature in that the introduction target substance is delivered from the supply unit of the delivery device, and the introduction target substance is supplied to a duct located upstream of the target site of the tissue existing in the living body. . Therefore, according to the method 1, it is possible to suppress the occurrence of unnecessary damage at the target site of the tissue.
- the method 1 has one major feature in that the pressure applied to the target site of the tissue is generated in the pressure load portion at the same time or after the supply of the substance to be introduced into the conduit. Therefore, according to the method 1, the introduction target substance can be locally delivered to the target site to which the pressure is applied. Further, even when a substance that does not easily penetrate the cell membrane is used as the introduction target substance, the substance can be delivered to the target site.
- Examples of the living body include animals having a conduit for transporting body fluids and metabolites.
- Examples of such animals include humans, non-human mammals, birds (eg, chickens, quails, etc.) and the like.
- FIG. 1 is a block diagram showing a configuration of a delivery target substance delivery device used in the present invention.
- a delivery device 1 shown in FIG. 1 supplies a pressure loading unit 10a that applies pressure to a target site of a tissue existing in a living body, and supplies a substance to be introduced to a conduit located upstream of the target site of the tissue. And a control unit 40 that controls the pressure load unit 10 a and the supply unit 30.
- the delivery device used in the method 1 is roughly classified into a device that applies a suction pressure as a pressure (first delivery device) and a device that loads a pressing force as a pressure (second delivery device). Hereinafter, each delivery device will be described.
- FIG. 2 is a schematic explanatory diagram showing a configuration of a main part of a delivery device (first delivery device) for a substance to be introduced used in the present invention.
- FIG. 3 is a schematic explanatory diagram showing the configuration of the pressure load section of the first delivery device.
- the first delivery device 1 includes a pressure load section 10a for loading the suction pressure to the target site S 1 organization S 0 present in the living body, the conduit located upstream of the target site S 1 of the tissue S 0 S 2 includes a supply unit 30 that supplies the substance to be introduced, and a control unit (not shown) that controls the supply unit 30 and the pressure load unit 10a.
- the pressure load unit 10 a is brought into contact with the tissue S 0 and connected to a suction unit 11 that sucks the target site S 1 , and a suction device that sucks air in the suction unit 11 and other devices.
- a bendable tube 13 that connects between the suction part 11 and the connection part 12.
- One end of the connection part 12 is connected to the tube body 13, and the other end can be connected to an aspirator that sucks out air in the suction part 11 and other devices (for example, an endoscope, forceps, etc.).
- the suction part 11 is cup-shaped and has a suction port 11a for sucking out air in the suction part 11 (see FIG. 2).
- the air in the suction portion 11 from the suction port 11a is that the sucked out, sucking the target site S 1 organization S 0 (see FIG. 2).
- the introduction target substance injected into the pipe line S 2 by the supply unit 30 is delivered to the target site S 1 .
- the size of the pressure load portion 10a is such a size that it can be attached to an aspirator for sucking out air in the suction portion 11 or other devices (for example, an endoscope, forceps, etc.).
- the supply unit 30 includes a storage unit 30a that stores the introduction target substance and a sending unit 30b that sends out the introduction target substance.
- Sending part 30b, as inserted into the pipe S 2 located upstream of the target site S 1 can be introduced introducing the substance into the pipe S 2, which is the tip and needle.
- the control unit 40 controls the pressure load unit 10 a and the storage unit 30 a and the delivery unit 30 b of the supply unit 30.
- the control unit 40 performs suction of the target site S 1 by the pressure load unit 10a, transfer of the introduction target substance from the storage unit 30a to the delivery unit 30b, delivery of the introduction target substance from the delivery unit 30b to the pipeline S 2, and the like. I have control.
- the delivery device 1 may be configured to manually control the pressure load unit 10a and the supply unit 30 instead of the control unit.
- the delivery device 1 for sucking the target site S 1 by the pressure load section 10a, by using a delivery device 1, as compared with the conventional direct injection method, it is possible to suppress damage to the target site S 1. Further, according to the delivery device 1 can be without blocking the blood flow as the conventional method using a catheter to deliver the introduction substance a suction pressure to a target site S 1 that was loaded.
- the delivery device 1 when the delivery of the introduction substance into such organs in vivo, installed insertion opening degree that can be inserted pressure load section 10a when brought into forming the living body, the pressure load section 10a to a target site S 1 can do. Therefore, according to the delivery device 1, it is possible to deliver the substance to be introduced with minimal invasiveness.
- FIG. 4 is a schematic explanatory view showing a configuration of a main part of a delivery device (second delivery device) for a substance to be introduced used in the present invention.
- Delivery device 2 shown in FIG. 4 the pressure load section 10b for loading the pressing force to a target site S 1 organization S 0 present in the living body, the tube located upstream of the target site S 1 of the tissue S 0 and a supply unit 30 for supplying introducing the substance into the road S 2.
- the delivery apparatus 2 has the control part 40 (personal computer) which controls the pressure load part 10b and the supply part 30.
- the fluid supply unit 25 includes a fluid storage unit 26 that stores fluid, a converter 27 that converts an electrical signal from the control unit 40 into a fluid pressure signal, a pressure sensor 28 that detects the pressure of the fluid, and a port that is punctured by a living body. 23, and a holding portion 25a that holds the needle portion 29.
- the converter 27 is connected to the control unit 40.
- the holding part 21 forms a gap through which the duct S 2 connected to the organ and the like passes through the side of the holding part main body 121 and the holding part main body 121 and the organ having the target site S 1.
- 121 includes a duct-side lid portion 122 that suppresses displacement of the organ and the like in the lateral direction of 121 and an upper lid portion 123 that suppresses displacement of the organ and the like in the vertical direction of the holding body 121 (FIG. 5). reference).
- the holding unit 21 can be embedded in the living body.
- the holding part main body 121 includes a bottom part 121a and side parts 121b, 121c, and 121d (see FIG. 5).
- a stopper 134 is formed on the inner side of the end portion 132 to fix the vertical position of the duct-side cover 122 and form a gap through which the duct S 2 leading to an organ or the like passes [FIG.
- a groove part 135 is formed in the longitudinal direction of each of the side parts 121b and 121c so that the upper lid part 123 can be fitted therein. See FIGS. 6A and 6C].
- the pressing portion 22 includes a pressing portion main body 22a, eight balloons 22b provided integrally with the pressing portion main body 22a, a flow path 22c for supplying fluid to the eight balloons 22b, And a fluid introduction port 22d for introducing a fluid.
- the balloon 22b is configured to expand when a fluid is supplied. Thereby, the introduction target substance injected into the duct (blood vessel) S 2 by the supply unit 30 is delivered to the target site S 1 (see FIG. 4).
- the size of the holding portion 21 and the port 23 of the pressure load portion 10b varies depending on, for example, the size of the organ in which the target site S1 exists, and thus the size of the organ in which the target site S1 exists. It is desirable to determine appropriately according to the above.
- the supply unit 30 is the same as the supply unit of the delivery device 1.
- the control unit 40 controls the pressure load unit 10b and the storage unit 30a and the delivery unit 30b of the supply unit 30.
- the control unit 40 controls the load of the pressing force by the pressure load unit 10b, the transfer of the introduction target substance from the storage unit 30a to the delivery unit 30b, the delivery of the introduction target substance from the delivery unit 30b, and the like.
- the delivery device 2 is connected to the kidney, liver, pancreas, stomach, spleen, heart, small intestine, large intestine, lung, uterus, bladder, esophagus, duodenum, brain, eyeball, trachea, skeletal muscle, diaphragm, adrenal gland, prostate, ovary,
- the holding unit 21 may have a shape suitable for holding the tissue.
- the delivery device 2 for the holding portion 21 can fix the position of the target site S 1, positional displacement when the load of the pressing force to the target site S 1 is less likely to occur. Therefore, by using the delivery device 2, the specificity at the time of delivery of the substance to be introduced can be improved.
- FIG. 8 is a process chart showing the processing procedure of the operating method of the delivery target substance delivery device according to the embodiment of the present invention.
- the pressure load section 10a of the delivery device 1 Prior to performing the method 1-1, the pressure load section 10a of the delivery device 1 with previously set the target site S 1 organization S 0 present in the living body, tissue S 0 present the supply portion 30 into the living body Is set in the pipeline S 2 located upstream of the target site S 1 (see “setting step S1-1” in FIG. 8)].
- method 1-1 first, sent the introduction substance from a supply 30, supplies the introduced substance to the conduit S 2 [step (1-1) (in FIG. 8, "introducing the substance supply step S1- 2 ”))].
- the supply position of the introduction target substance in the pipeline S 2 can be appropriately set according to the type and stability of the introduction target substance, the position of the target site S 1 , and the like.
- the supply position of the introduction target substance is such that the distance between the supply position of the introduction target substance and the target site S 1 is such that the nucleic acid is not decomposed, and It is preferable that the position be easy to supply.
- the supply amount of the introduction substance, the type of the introduction substance, differs by the kind of the target site S 1, can not be determined unconditionally. Accordingly, the supply amount of the introduction substance, the type of the introduction substance, it is preferably determined according to the type of target site S 1.
- the suction pressure loading step S1-3 may be performed simultaneously with the introduction target substance supply step S1-2, or may be performed after the introduction target substance supply step S1-2. Timing of such suction pressure loading step S1-3 may be appropriately set depending on the distance between the supply position and the target site S 1 of the introduction substance.
- the introduction target substance can be specifically delivered to the target site S 1 .
- FIG. 9 is a process chart showing a processing procedure of a method for delivering a substance to be introduced according to another embodiment of the present invention.
- the pressure load section 10b of the delivery device 2 together with the previously set target site S 1 organization S 0 present in the living body, tissue S 0 present the supply portion 30 into the living body Is set in the pipeline S 2 located upstream of the target site S 1 (see (A) in FIG. 9).
- tissue S 0 present the supply portion 30 into the living body Is set in the pipeline S 2 located upstream of the target site S 1 (see (A) in FIG. 9).
- the organ in which the target site S 1 exists is accommodated in the holding unit 21 and the pressing unit 22 is arranged so as to surround the organ.
- method 1-2 first, it sent the introduction substance from a supply 30 (also referred to as "introduction substance supplying step") supplying the introduced substance to the conduit S 2 [step (1-1), FIG. 9 (see (B)).
- a supply 30 also referred to as "introduction substance supplying step” supplying the introduced substance to the conduit S 2 [step (1-1), FIG. 9 (see (B)).
- the supply amount of the introduction substance as in the case of the method 1-1, the supply amount of the introduction substance, the type of the introduction substance, it is preferably determined according to the type of target site S 1. Further, the supply position of the substance to be introduced in the pipeline S 2 should be appropriately set according to the type and stability of the substance to be introduced, the position of the target site S 1 and the like, as in the case of the method 1-1. Can do.
- the amount of fluid supplied to the balloon 22b is proportional to the pressing force to a target site S 1. Then, the pressing force to a target site S 1 is proportional to the amount of introduced substance to be introduced to the target site S 1. Accordingly, the pressing force loading step, the amount of fluid supplied to the balloon 22b is preferably determined according to the amount of introduction the substance to be introduced to the target site S 1. In general, it is preferable to adjust the amount of fluid so that the pressure is 30 kPa to 80 kPa.
- the pressing force loading step may be performed simultaneously with the introduction target substance supply step or after the introduction target substance supply step.
- the timing for performing the pressing force loading step can be appropriately set according to the distance between the supply position of the introduction target substance and the target site S 1 .
- a method for delivering a substance to be introduced comprising: (2-1) supplying a substance to be introduced to a conduit located upstream of the tissue, and (2-2) simultaneously with the step (2-1) or after the step (2-1),
- the present invention relates to a method for delivering a substance to be introduced, which includes a step of applying pressure to a target site of tissue and delivering the substance to be introduced to the target site (hereinafter referred to as “method 2”).
- Method 2 includes the step of supplying the substance to be introduced into a duct located upstream of the target site of the tissue of the non-human mammal and the target site of the tissue at the same time or after the feeding of the substance to be introduced into the duct. There is one major feature in the point of loading the pressure against. Therefore, the method 2 has the same effect as the method 1.
- non-human mammals examples include monkeys, pigs, dogs, horses, mice, rabbits, rats, cows, sheep, guinea pigs, and hamsters.
- Method 2 includes a method using suction pressure (referred to as “method 2-1”) as the pressure applied to the target site and a method using pressure (referred to as “method 2-2”) as the pressure applied to the target site. Broadly divided.
- the method 2-1 can be carried out in the same manner as the method 1-1, for example, by using the delivery device 1 shown in FIG. 1 and performing the steps shown in FIG. Therefore, set if, prior to the method 2-1, at least the suction portion 11 of the pressure load section 10a of the delivery device 1 to the target site S 1 organization S 0 present in the living body using a delivery device 1 shown in FIG. 1
- the supply unit 30 is set in the duct S 2 positioned upstream of the target site S 1 of the tissue S 0 existing in the living body (see “setting step S1-1” in FIG. 8). .
- step (2-1) to supply the introduced substance to the conduit S 2 located upstream of the tissue S 0 of a non-human mammal [step (2-1), in FIG. 8, "introducing the substance supply step Refer to S1-2].
- the supply amount of the introduction substance, the supply amount of the introduction substance, the type of the introduction substance, it is preferably determined according to the type of target site S 1.
- Supply position of the introduction substance in the tube path, the type and stability of the introduced substance, depending on the position of the target site S 1, can be set as appropriate.
- the supply position of the introduction target substance is such that the distance between the supply position of the introduction target substance and the target site S 1 is a distance that can stably hold the introduction target substance, and the supply of the introduction target substance is easy. It is preferable that
- the supply amount of the introduction substance, the type of introduced substance can be determined according to the type of target site S 1.
- a pressure is applied to the target site S 1 of the tissue S 0 and introduced into the target site S 1 .
- the target substance is delivered [see step (2-2), “suction pressure loading step S1-3” in FIG. 8].
- the size of the suction pressure is preferably determined in accordance with the delivery amount of the introduced substance to a target site S 1. Usually, it is 10 kPa to 80 kPa.
- step (2-2) aspiration target site S 1 in step (2-2), step (2-1) at the same time made even better, may be performed after the step (2-1). Timing of this step (2-2) can be appropriately set depending on the distance between the supply position and the target site S 1 of the introduction substance.
- the method 2-2 can be performed by performing the steps shown in FIG. 9 using the delivery device 2 shown in FIG. 4, for example, as in the method 1-2. Therefore, when using the delivery device 2 shown in FIG. 4, prior to the method 2-2, the pressure load unit 10b of the delivery device 2 is set at the target site S 1 of the tissue S 0 existing in the living body, The supply unit 30 is set in the duct S 2 located upstream of the target site S 1 of the tissue S 0 existing in the living body (see (A) in FIG. 9). At this time, the organ in which the target site S 1 exists is accommodated in the holding unit 21 and the pressing unit 22 is arranged so as to surround the organ.
- the substance to be introduced is supplied to the pipeline S 2 located upstream of the tissue S 0 (see (B) in FIG. 9).
- the supply amount of the introduction substance, the supply amount of the introduction substance, the type of the introduction substance, it is preferably determined according to the type of target site S 1.
- the supply position of the introduction target substance in the pipeline S 2 can be appropriately set according to the type and stability of the introduction target substance, the position of the target site S 1 , and the like.
- the supply position of the introduction target substance is such that the distance between the supply position of the introduction target substance and the target site S 1 is a distance that can stably hold the introduction target substance, and the supply of the introduction target substance is easy. It is preferable that
- the target site S 1 is pressed while fixing the position of the tissue S 0 [see (C) in FIG. 9].
- a pressing force to a target site S 1, capable of delivering the introduced substance to a target site S 1.
- the fixed position of the tissue S 0, when the tissue S 0 is a tissue of an organ the step (2-2), wherein to accommodate the organs in a case that can house the organ tissue It is preferable to press the target site S 1 while fixing the position of S 0 .
- the magnitude of the pressing force to a target site S 1 is preferably determined according to the amount of introduction the substance to be introduced to the target site S 1. Usually, it is 30 kPa to 80 kPa.
- the pressing to the target site S 1 in the step (2-2) may be performed simultaneously with the step (2-1), or may be performed after the step (2-1). Timing of this step (2-2) can be appropriately set depending on the distance between the supply position and the target site S 1 of the introduction substance.
- the delivery method of the substance to be introduced can be applied to humans in addition to non-human animals.
- nucleic acids, drugs, peptide compounds, antibodies, polymer compounds, metals (for example, metal nanoparticles), semiconductors (for example, quantum dots), or a combination of these in the treatment and prevention of human diseases It is possible to locally deliver various introduction target substances such as the introduced substance to the target site, local delivery to the target site of the introduction target substance in human diagnosis, and the like.
- the present invention provides a method for delivering a substance to be introduced that delivers a target substance to be introduced to a target site of a target derived from a living body. Because (3-1) supplying the substance to be introduced to a site different from the target site in the target, and (3-2) simultaneously with the step (3-1) or after the step (3-1) , Applying a pressure to the target site of the object and delivering the substance to be introduced to the target site;
- the present invention relates to a method for delivering a substance to be introduced to a target site of a target object (hereinafter referred to as “method 3”).
- Method 3 is a method in which the introduction target substance is supplied to a site different from the target site of the target derived from the living body and at the same time or after the supply of the target substance to the site different from the target site of the target derived from the living body.
- One major feature is that it applies pressure to the target site of the tissue. Therefore, the method 3 has the same effects as the method 1.
- Examples of the living body include humans, non-human mammals, and birds (for example, chickens and quails).
- Examples of the living body-derived object include tissues isolated from the living body (for example, kidney, liver, pancreas, stomach, spleen, heart, small intestine, large intestine, lung, uterus, bladder, esophagus, duodenum, brain, eyeball, trachea, Skeletal muscle, diaphragm, adrenal gland, prostate, ovary, fallopian tube, etc.), organs isolated from living bodies (eg, kidney, liver, pancreas, stomach, spleen, heart, small intestine, large intestine, lung, uterus, bladder) , Esophagus, duodenum, brain, eyeball, trachea, skeletal muscle, diaphragm, adrenal gland, prostate, ovary, fallopian tube, etc.), artificial tissue composed of living cells (for example, single type derived from somatic cells and stem cells) Cell aggregates composed of cells, aggregates composed of a single type of cell population derived from somatic cells and stem cells and cell scaffolds,
- Method 3 includes a method using suction pressure (referred to as “Method 3-1”) as the pressure applied to the target site and a method using pressure (referred to as “Method 3-2”) as the pressure applied to the target site. Broadly divided.
- the method 3-1 can be performed, for example, by using the delivery device 1 shown in FIG. 1 and performing the steps shown in FIG. 8 in the same manner as the methods 1-1 and 2-1. Therefore, when using the delivery device 1 shown in FIG. 1, prior to the method 3-1, at least the suction portion 11 of the pressure load portion 10a of the delivery device 1 is set at the target site S1, and the supply portion 30 is set in a site different from the labeled site S 1 (see “setting step S1-1” in FIG. 8).
- method 3-1 first, in the object, and it supplies the introduced substance to a site different from the labeled site S 1 [step (3-1), in FIG. 8, "introducing the substance supply step S1-2" reference ⁇ .
- the supply amount of the introduction substance, the supply amount of the introduction substance, the type of the introduction substance, it is preferably determined according to the type of target site S 1.
- Supply position of the introduction substance into the object, the type and stability of the introduced substance, depending on the position of the target site S 1, can be set as appropriate.
- the supply position of the introduction target substance is such that the distance between the supply position of the introduction target substance and the target site S 1 is a distance that can stably hold the introduction target substance, and the supply of the introduction target substance is easy. It is preferable that
- the supply amount of the introduction substance, the type of introduced substance can be determined according to the type of target site S 1.
- step (3-1) After said step (3-1) and simultaneously or the step (3-1), and the load pressure (suction pressure) to a target site S 1 of the object, introducing the object into the target site S 1
- the substance is delivered [step (3-2) (see “suction pressure loading step S1-3” in FIG. 8)].
- the size of the suction pressure is preferably determined in accordance with the delivery amount of the introduced substance to a target site S 1. Usually, it is 10 kPa to 80 kPa.
- step (3-1) at the same time made even better may be performed after the step (3-1).
- the timing of performing the step (3-2) can be appropriately set according to the distance between the supply position of the introduction target substance and the target site S 1 .
- the method 3-2 is suitable for a case where a tissue such as muscle isolated from a living body, an organ isolated from a living body, or an artificial organ composed of living body cells is used as an object derived from a living body.
- the method 3-2 can be carried out, for example, by using the delivery device 2 shown in FIG. 4 and performing the steps shown in FIG.
- the pressure load section 10b of the delivery device 2 together with the previously set target site S 1 of the object derived from a living organism, in the object supply unit 30, the target site S 1 They are set in different parts (see (A) in FIG. 9).
- the introduction target substance is supplied to a site different from the target site S 1 (see (B) in FIG. 9).
- the supply amount of the introduction substance, the supply amount of the introduction substance, the type of the introduction substance, it is preferably determined according to the type of target site S 1.
- Supply position of the introduction substance in the object, from the viewpoint of improving the efficiency of delivery of the introduction substance is preferably a site of transport of substances is carried out between the target site S 1.
- Examples of the site where the substance is transported to and from the target site S 1 include a pipeline, an artificially created pipeline, and the like.
- the supply position of the introduction substance, the type and stability of the introduced substance is preferably set depending on the position of the target site S 1.
- the supply position of the introduction target substance is such that the distance between the supply position of the introduction target substance and the target site S 1 is such that the introduction target substance can be stably held, and the supply of the introduction target substance is performed. It is preferable that the position is easy.
- step (3-2) pressing is performed while fixing the position of the target site S 1 [see (C) in FIG. 9].
- a pressing force to a target site S 1, capable of delivering the introduced substance to a target site S 1.
- the target site S 1 is immobilized on a tissue such as muscle isolated from a living body, an organ isolated from a living body, or an artificial organ composed of cells of a living body, In 3-2), this is performed by housing the organ in a case capable of housing the organ and restraining the organ.
- the magnitude of the pressing force to a target site S 1 is preferably determined according to the amount of introduction the substance to be introduced to the target site S 1. Usually, it is 30 kPa to 80 kPa.
- step (3-2) the pressing to the target site S 1 in step (3-2), step (3-1) at the same time made even better, may be performed after the step (3-1).
- the timing of performing the step (3-2) can be appropriately set according to the distance between the supply position of the introduction target substance and the target site S 1 .
- the introduction target substance can be efficiently delivered locally to the target site. Therefore, the operation method of the delivery target substance delivery device and the delivery target substance delivery method of the present invention include nucleic acids, drugs, peptide compounds, antibodies, polymer compounds, metals (for example, metal nanoparticles, etc.) in the treatment and prevention of diseases. ), Semiconductor (for example, quantum dots, etc.), local delivery to the target site of various introduction target substances such as substances composed of them, and local introduction of the introduction target substance to the target site in diagnosis It is extremely useful for effective delivery.
- Example 1 The abdominal cavity of an anesthetized mouse was dissected along the midline to expose the liver. Then, the suction part 11 (refer FIG. 2 and FIG. 3) of the pressure load part 10a of the delivery apparatus 1 of the introduction object substance was installed in the target site
- an outer diameter D 2 is 5 mm
- the height h of the inner wall is 3 mm.
- the internal diameter d of the suction port 11a of the suction part 11 is 1 mm.
- the plasmid pCMV-LUC is a plasmid prepared from a plasmid carrying a nucleic acid encoding luciferase (GenBank accession number: U47296, pGL3-control manufactured by Promega) [T.
- FIG. 10 shows the results of examining the expression site of luciferase encoded by the injected nucleic acid in Example 1 in Example 1.
- the site indicated by the arrow indicates the site where luminescence based on luciferase was observed.
- Example 2 In Example 1, after administration of a solution containing luciferin, the abdominal cavity was opened and the liver of the mouse was removed. The same operation as in Example 1 was performed, and luminescence based on luciferase was photographed with a camera and observed with the naked eye. (Example 2).
- FIG. 11 shows the results of examining the expression site of the luciferase encoded by the injected nucleic acid when one target site in the mouse liver was aspirated in Example 2. In the figure, (A) shows the result of direct observation of the mouse liver, and (B) shows the result of examining the expression site of luciferase.
- Example 2 the same operation as in Example 2 was performed except that two target sites in the liver of the mouse were simultaneously aspirated, and luminescence based on luciferase was photographed with a camera and observed with the naked eye (implementation) Example 3).
- FIG. 12 shows the results of examining the expression site of luciferase encoded by the injected nucleic acid in Example 3 when two target sites in the mouse liver were aspirated.
- (A) shows the result of direct observation of the mouse liver
- (B) shows the result of examining the expression site of luciferase.
- Example 1 In Example 1, except that the nucleic acid was injected under various conditions shown in Table 1, the same operation as in Example 1 was performed, and luminescence based on luciferase was photographed with a camera and observed with the naked eye.
- the diameter D 1 is the inner diameter D 1 of the suction part 11 of the pressure load part 10a of the delivery device 1 shown in FIG.
- FIG. 13 shows the results of examining the expression site of luciferase encoded by nucleic acid injected under various conditions in Test Example 1.
- nucleic acid can be more efficiently delivered to the target site by sucking the target site using the delivery device having the suction unit 11.
- Example 4 The abdominal cavity of an anesthetized mouse was dissected along the midline to expose the kidney. Then, the suction part 11 (refer FIG. 2 and FIG. 3) of the pressure load part 10a of the delivery apparatus 1 was installed in the target site
- the mouse was euthanized under anesthesia, the kidney was taken out, and the mass was measured. Thereafter, the kidney is set to 5 mL of Lysis solution per 1 g of kidney with respect to the Lysis solution [composition: 0.05 mass% Triton (registered trademark) X-100, 2 mM ethylenediaminetetraacetic acid, 0.1 M Tris-hydrochloric acid]. Added. And the obtained mixture was homogeneously soluted at 4 degreeC using the homogenizer, and the tissue crushing liquid was obtained. Next, the tissue disruption solution was subjected to centrifugation (10000 ⁇ g, 10 minutes, 4 ° C.) to obtain a supernatant.
- Lysis solution Composition: 0.05 mass% Triton (registered trademark) X-100, 2 mM ethylenediaminetetraacetic acid, 0.1 M Tris-hydrochloric acid.
- the luciferase activity when the suction part having the inner diameter D 1 of 4 mm and the outer diameter D 2 of 6 mm is used is the inner diameter D 1 of 2 mm and the outer diameter D. It can be seen that 2 is higher than the luciferase activity when using a suction part of 3 mm. Therefore, this result suggests that the amount of nucleic acid to be delivered can be set by adjusting the size of the suction part.
- the number of sites to which the suction pressure is applied is one site for the luciferase activity when the number of sites to which the suction pressure is applied is three. It can be seen that it is higher than the luciferase activity. Therefore, this result suggests that the amount of nucleic acid to be delivered or the location to be delivered can be set by repeating aspiration.
- Test Example 2 The abdominal cavity of an anesthetized mouse was dissected along the midline to expose the liver. Then, the suction part 11 (refer FIG. 2 and FIG. 3) of the pressure load part 10a of the delivery apparatus 1 was installed in the target site
- the position in the liver of the part where the suction part is installed is shown in FIG. In the figure, the arrow corresponds to the portion where the suction port is installed. A broken line portion corresponds to a portion where the wall portion of the suction portion is in contact. Further, FIG. 15B shows an enlarged explanatory view of the installation portion of the suction part shown in FIG.
- FIG. 15C shows a section of a portion to be measured for luciferase activity in Test Example 2.
- FIG. 16 shows the results of examining the relationship between the liver site and the relative activity of luciferase in Test Example 2.
- FIG. 16 shows that the luciferase activity is highest in the portion (section 1) up to a depth of about 1.5 mm among the portions in contact with the suction port. This result suggested that the nucleic acid was locally delivered to the surface of the aspiration site.
- 18 ⁇ g of pCMV-Luc alone (introduced sample number 1), 18 ⁇ g of pCMV-Luc and 10 ⁇ g of siRNA (siRNA-LUC) for nucleic acid encoding luciferase are supplied to the tail blood vessel of the delivery device 1 by the delivery device 1.
- a combination (introduced sample number 2) or a combination of 18 ⁇ g of pCMV-Luc and 10 ⁇ g of scrambled siRNA (introduced sample number 3) was injected.
- the target site in the liver was aspirated at 80 kPa for 5 seconds by the pressure load unit 10a of the delivery device 1. Thereafter, the abdominal cavity of the mouse was closed and the mouse was raised for 6 hours.
- the mouse was euthanized under anesthesia, the liver was taken out, and the mass was measured. Thereafter, the liver was added to the Lysis solution so that the Lysis solution was 25 mL per 1 g of the liver.
- the resulting mixture was homogenized at 4 ° C. using a homogenizer to obtain a tissue disruption solution.
- the tissue disruption solution was subjected to centrifugation (10000 ⁇ g, 10 minutes, 4 ° C.) to obtain a supernatant. 100 ⁇ L of the solution containing the luciferin was added to 10 ⁇ L of the obtained supernatant and mixed to obtain a mixture. And the luciferase activity of the said mixture was measured using the said luminescence amount determination apparatus.
- FIG. 17 shows the result of examining the relationship between the type of the introduced sample and the relative activity of luciferase in Test Example 3.
- results shown in FIG. 17 indicate that the relative luciferase activity when using a combination of pCMV-Luc and siRNA-LUC is 17% of the relative luciferase activity when using pCMV-Luc alone.
- the nucleic acid injected into the target different from the target site in the object is locally delivered to the target site to which pressure (suction pressure) is applied, and at the target site, siRNA It can be seen that the function (RNA interference) is expressed.
- the pressing unit 22 includes a pressing unit main body 22a, four balloons 22b provided integrally with the pressing unit main body 22a, and an air supply unit 22d connected to each of the four balloons 22b via a flow path. (See FIG. 18).
- the mouse was euthanized under anesthesia, the kidney was taken out, and the mass was measured. Thereafter, using a homogenizer, the kidney was added to the Lysis solution so as to be 5 mL of Lysis solution per 1 g of kidney. The resulting mixture was homogenized at 4 ° C. using a homogenizer to obtain a tissue disruption solution. Next, the tissue disruption solution was subjected to centrifugation (10000 ⁇ g, 10 minutes, 4 ° C.) to obtain a supernatant. To 10 ⁇ L of the obtained supernatant, 100 ⁇ L of the solution containing the luciferin was added and mixed to obtain a mixture.
- FIG. 19 shows the results of examining the relationship between the load air pressure or pressure load technique and luciferase activity in Test Example 4.
- FIG. 19 shows that the luciferase activity can be detected in the mouse kidney. Therefore, from this result, the nucleic acid injected into the target in the target object different from the target site is locally delivered to the target site loaded with pressure (pressing force), and in the target site, It can be seen that the protein encoded by the nucleic acid is expressed.
- the results shown in FIG. 19 indicate that the luciferase activity increases as the size of the load air pressure increases.
- luciferase activity can be detected, but it is difficult to adjust the expression level.
- fixed and fixed by the plate it will only touch a tissue surface with a balloon and it will be difficult to deliver a nucleic acid. Therefore, this result suggests that the amount of nucleic acid to be delivered (expression amount) can be set by adjusting the magnitude of the load air pressure.
- the abdominal cavity of the mouse was closed so that only a part of the pressure load portion 22 (portion 22d in FIG. 7) was exposed outside the abdominal cavity of the mouse.
- 100 ⁇ g of the pCMV-Luc was injected into the tail blood vessel of the mouse by the supply unit 30 of the delivery device 2.
- the kidney was pressed at 60 kPa (load air pressure) for 5 seconds by the pressing portion 22 of the delivery device 2. Thereafter, the mice were raised for 6 hours.
- the mouse was euthanized under anesthesia, the kidney was taken out, and the mass was measured. Thereafter, using a homogenizer, the kidney was added to the Lysis solution so as to be 5 mL of Lysis solution per 1 g of kidney.
- the kidney in the obtained mixture was homogeneously soluted at 4 ° C. using a homogenizer to obtain a tissue disruption solution.
- the tissue disruption solution was subjected to centrifugation (10000 ⁇ g, 10 minutes, 4 ° C.) to obtain a supernatant. 100 ⁇ L of the solution containing luciferin was added to 10 ⁇ L of the obtained supernatant to obtain a mixture.
- FIG. 20 shows the result of examining the relationship between the pressure load condition and the luciferase activity in Test Example 5.
- the target substance such as nucleic acid is introduced by injecting the target substance such as nucleic acid into a part different from the target part and applying pressure (suction pressure or pressing force) to the target part. It can be delivered locally to the target site, and the function of the substance to be introduced such as the nucleic acid at the target site (for example, when the substance to be introduced is a nucleic acid, expression of the protein encoded by the nucleic acid, nucleic acid It can be seen that the above functions can be expressed.
- a pressing force is applied to the target site, it can be seen that the introduction target substance such as a nucleic acid can be efficiently delivered to the target site by fixing the target site with the holding portion.
- the present invention is useful for local delivery to target sites such as nucleic acid drugs.
- Example 5 In Example 4, except that the kidney, heart, spleen, or liver was used as the object, and the conditions of the pressure load unit 10a installed at the target site in the object were the conditions shown in Table 3, Example 4 The same operation as above was performed, and luciferase activity was measured.
- FIG. 21 shows the result of examining the relationship between the type of the object and the luciferase activity in Example 5.
- luciferase activity is detected in any of the kidney, heart, spleen and liver. Therefore, from these results, in any of the kidney, heart, spleen and liver, the nucleic acid injected into a site different from the target site is locally delivered to the target site loaded with pressure (suction pressure). Moreover, it can be seen that the protein encoded by the nucleic acid is expressed at the target site.
- Example 6 Example 6
- Example 4 was used except that the heart was used instead of the kidney as the object, and each of the delivery devices of device numbers 5 to 7 shown in Table 4 was used as the delivery device 1. The same operation was performed and luciferase activity was measured. The results of examining the relationship between the type of device used and luciferase activity in Example 6 are shown in FIG.
- the luciferase activity when the suction part having the inner diameter D 1 of 3 mm and the outer diameter D 2 of 4 mm is used is that the inner diameter D 1 is 2 mm. and it can be seen that the outer diameter D 2 is compared with the luciferase activity when using a suction unit of 3 mm, is high. Therefore, this result suggests that the amount of nucleic acid to be delivered can be set by adjusting the size of the suction part.
- Test Example 6 The abdominal cavity of an anesthetized mouse was dissected along the midline to expose the kidney. Then, the pressure load part 10a (refer FIG. 2 and FIG. 3) of the delivery apparatus 1 was installed in the target site
- Bovine serum albumin was added to a phosphate buffer solution (PBS) so that its concentration was 0.1% by mass to obtain a 0.1% by mass BSA-PBS solution.
- a firefly luciferase enzyme solution [manufactured by Toyo B-Net Co., Ltd.] was added to the obtained 0.1% by mass BSA-PBS solution so that the concentration of firefly luciferase was 0.04 ⁇ g / 200 ⁇ L to obtain a mixture.
- the mixture was injected into the tail blood vessel of the mouse by the supply unit 30 of the delivery device 1. Immediately thereafter, a suction pressure was applied to the target site in one of the kidneys under the conditions shown in Table 5 for 5 seconds by the pressure loading unit 10a of the delivery device 1. Immediately thereafter, luciferase activity was measured.
- mice were exsanguinated under anesthesia and euthanized, and then both kidneys were taken out and weighed.
- the kidney having the target site loaded with the suction pressure was added to the Lysis solution so that the Lysis solution was 10 mL per 1 g of the kidney. And the obtained mixture was homogeneously soluted at 4 degreeC using the homogenizer, and the tissue crushing liquid was obtained.
- the tissue disruption solution was subjected to centrifugation (10000 ⁇ g, 10 minutes, 4 ° C.) to obtain a supernatant.
- 100 ⁇ L of the solution containing the luciferin was added and mixed to obtain a mixture.
- the luciferase activity of the said mixture was measured using the light-emission quantity determination apparatus [EG & G BERTHOLD company make, brand name: Lumat LB 9507] (introduction method of experiment number 1).
- luciferase activity was measured in the same manner as described above except that a kidney not loaded with suction pressure was used instead of the kidney having the target site loaded with suction pressure (the experiment number 2). Introduction method).
- FIG. 23 shows the result of examining the relationship between the type of introduction method and the relative activity of luciferase in Test Example 6.
- the luciferase activity when the introduction method of experiment number 1 is performed is higher than the luciferase activity when the method of experiment number 2 is performed. Therefore, from this result, the protein injected into a site different from the target site is locally delivered to the target site loaded with pressure (suction pressure), and the physiological state of the protein at the target site. It can be seen that the activity is expressed.
- a target substance such as a nucleic acid or a peptide compound is injected into a part different from the target part, and pressure (suction pressure or pressing force) is applied to the target part.
- the substance can be locally delivered to the target site and that the function of the substance to be introduced can be expressed at the target site. Therefore, the present invention provides nucleic acids, drugs, peptide compounds, antibodies, polymer compounds, metals (for example, metal nanoparticles), semiconductors (for example, quantum dots), or a combination thereof in the treatment and prevention of diseases. It is suggested that the present invention is useful for local delivery of various introduction target substances such as configured substances to a target site, local delivery of the introduction target substance to a target site in diagnosis, and the like.
- a mixture obtained by adding a drug, antibody, polymer compound, metal nanoparticle or quantum dot to a solvent is injected into a site different from the target site, and a suction pressure or a pressing force is applied to the target site.
- a drug, an antibody, a high molecular compound, a metal nanoparticle, or a quantum dot can be locally delivered to a target site.
- the presence of a drug, antibody, polymer compound, metal nanoparticle or quantum dot at the target site can be confirmed by a detection method corresponding to each substance.
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Abstract
Description
また、前記特許文献1、非特許文献1および2に記載の方法では、カテーテルにより隔絶された領域には核酸を送達することができるが、臓器などにおける標的部位のみに局所的に核酸を送達したり、複数の標的部位に一度に核酸を送達したりすることが困難であるという欠点がある。
〔1〕 生体内に存在する組織の標的部位の上流に位置する管路に導入対象物質を供給する供給部と、前記組織の標的部位に圧力を負荷する圧力負荷部とを備えた導入対象物質の送達装置の作動方法であって、
(1-1)供給部から導入対象物質を送出させ、前記管路に前記導入対象物質を供給する工程、および
(1-2)前記工程(1-1)と同時または前記工程(1-1)の後、圧力負荷部に前記組織の標的部位に対する圧力を発生させる工程
を含む導入対象物質の送達装置の作動方法、
〔2〕 前記導入対象物質が核酸またはペプチド化合物である前記〔1〕に記載の方法、
〔3〕 前記圧力負荷部が、標的部位を吸引する吸引部を備えており、
前記工程(1-2)において、前記吸引部に前記標的部位を吸引させることにより、この標的部位に対する吸引圧を発生させる前記〔1〕または〔2〕に記載の方法、
〔4〕 前記圧力負荷部が、標的部位を押圧する押圧部と、前記組織の位置を固定して保持する保持部とを備えており、
前記工程(1-2)において、前記押圧部に前記標的部位を押圧させることにより、この標的部位に対する押圧力を発生させる前記〔1〕または〔2〕に記載の方法、
〔5〕 前記保持部が、生体内に埋め込み可能とされている前記〔4〕に記載の方法、
〔6〕 非ヒト哺乳動物の組織の標的部位に導入対象物質を送達する導入対象物質の送達方法であって、
(2-1) 前記組織の上流に位置する管路に導入対象物質を供給する工程、および
(2-2) 前記工程(2-1)と同時または前記工程(2-1)の後、前記組織の標的部位に圧力を負荷して、当該標的部位に前記導入対象物質を送達する工程
を含む導入対象物質の送達方法、
〔7〕 前記導入対象物質が核酸である前記〔6〕に記載の方法、
〔8〕 前記工程(2-2)において、標的部位に対して吸引圧を負荷する前記〔6〕または〔7〕に記載の方法、
〔9〕 前記工程(2-2)において、組織の位置を固定しながら、標的部位を押圧する前記〔6〕または〔7〕に記載の方法、
〔10〕 前記組織が臓器の組織であり、
前記工程(2-2)において、前記臓器を収容可能なケースに臓器を収容して前記組織の位置を固定しながら、標的部位を押圧する前記〔9〕に記載の方法、
〔11〕 生体由来の対象物の標的部位に導入対象物質を送達する導入対象物質の送達方法であって、
(3-1) 前記対象物において、標的部位とは異なる部位に導入対象物質を供給する工程、および
(3-2) 前記工程(3-1)と同時または前記工程(3-1)の後、前記対象物の標的部位に圧力を負荷して、当該標的部位に導入対象物質を送達する工程、
を含む対象物の標的部位への導入対象物質の送達方法、
〔12〕 前記導入対象物質が核酸またはペプチド化合物である前記〔11〕に記載の方法、
〔13〕 前記工程(3-2)において、標的部位に対して吸引圧を負荷する前記〔11〕または〔12〕に記載の方法、ならびに
〔14〕 前記工程(3-2)において、標的部位の位置を固定しながら、押圧する前記〔11〕または〔12〕に記載の方法
に関する。
本発明は、1つの側面では、生体内に存在する組織の標的部位の上流に位置する管路に導入対象物質を供給する供給部と、前記組織の標的部位に圧力を負荷する圧力負荷部とを備えた導入対象物質の送達装置の作動方法であって、
(1-1)供給部から導入対象物質を送出させ、前記管路に前記導入対象物質を供給する工程、および
(1-2)前記工程(1-1)と同時または前記工程(1-1)の後、圧力負荷部に前記組織の標的部位に対する圧力を発生させる工程
を含む導入対象物質の送達装置の作動方法に関する(以下、「方法1」という)。
〔1〕非ヒト哺乳動物の組織の標的部位への導入対象物質の送達方法
本発明は、他の側面は、非ヒト哺乳動物の組織の標的部位に導入対象物質を送達する導入対象物質の送達方法であって、
(2-1) 前記組織の上流に位置する管路に導入対象物質を供給する工程、および
(2-2) 前記工程(2-1)と同時または前記工程(2-1)の後、前記組織の標的部位に圧力を負荷して、当該標的部位に導入対象物質を送達する工程
を含む導入対象物質の送達方法に関する(以下、「方法2」という)。
本発明は、さらに他の側面では、生体由来の対象物の標的部位に導入対象物質を送達する導入対象物質の送達方法であって、
(3-1) 前記対象物において、標的部位とは異なる部位に導入対象物質を供給する工程、および
(3-2) 前記工程(3-1)と同時または前記工程(3-1)の後、前記対象物の標的部位に圧力を負荷して、当該標的部位に導入対象物質を送達する工程、
を含む対象物の標的部位への導入対象物質の送達方法に関する(以下、「方法3」という)。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、肝臓を露出させた。その後、導入対象物質の送達装置1の圧力負荷部10aの吸引部11(図2および図3参照)を肝臓における標的部位に設置した。なお、圧力負荷部10aの吸引部11の内径D1は3mm、外径D2は5mm、内壁の高さhは3mmである。また、吸引部11の吸引口11aの内径dは1mmである。
実施例1において、ルシフェリンを含む溶液を投与後、腹腔を開き、マウスの肝臓を取り出したことを除き、実施例1と同様に操作を行ない、ルシフェラーゼに基づく発光をカメラで撮影し、肉眼で観察した(実施例2)。実施例2において、マウスの肝臓における1ヶ所の標的部位を吸引した場合における注入した核酸にコードされたルシフェラーゼの発現部位を調べた結果を図11に示す。図中、(A)はマウスの肝臓を直接観察した結果、(B)はルシフェラーゼの発現部位を調べた結果を示す。
実施例1において、表1に示された種々の条件下に核酸を注入したことを除き、実施例1と同様に操作を行ない、ルシフェラーゼに基づく発光をカメラで撮影し、肉眼で観察した。なお、表中、直径D1は図2に示された送達装置1の圧力負荷部10aの吸引部11の内径D1である。試験例1において、種々の条件下に注入した核酸にコードされたルシフェラーゼの発現部位を調べた結果を図13に示す。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、腎臓を露出させた。その後、送達装置1の圧力負荷部10aの吸引部11(図2および図3参照)を腎臓における標的部位に設置した。なお、送達装置1として、表2に示されたデバイス番号:1~4の送達装置それぞれを用いた。吸引部11の内壁の高さhは、3mmである。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、肝臓を露出させた。その後、送達装置1の圧力負荷部10aの吸引部11(図2および図3参照)を肝臓における標的部位に設置した。なお、圧力負荷部10aの吸引部11の内径D1は4mm、外径D2は8mm、内壁の高さhは3mmである。試験例2において、吸引部を設置した部分の肝臓中における位置を図15(A)に示す。図中、矢印は、吸引口が設置された部分に対応する。また、破線部は、吸引部の壁部が接触した部分に対応する。また、図15(A)に示された吸引部の設置部分の拡大説明図を図15(B)に示す。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、肝臓を露出させた。その後、送達装置1の圧力負荷部10aの吸引部11(図2および図3参照)を肝臓における標的部位に設置した。なお、圧力負荷部10aの吸引部11の内径D1は4mm、外径D2は6mm、内壁の高さhは3mmである。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、腎臓を露出させた。つぎに、図18に示される押圧部22を腎臓に接触させ、これらを2枚のプレートで挟んで固定した。なお、押圧部22は、押圧部本体22aと、この押圧部本体22aに一体に設けられた4つのバルーン22bと、4つのバルーン22bそれぞれに流路を介して接続された空気供給部22dとを備えている(図18参照)。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、腎臓を露出させた。つぎに、図4に示される送達装置2の保持部21を構成する保持部本体121内に腎臓S0をセットした〔図6(A)参照〕。その後、血管側蓋部122を保持部本体121に取り付けた〔図6(B)参照〕。このとき、保持部本体121と血管側蓋部122との間の隙間から血管S2を保持部本体121外に出るようにした。つぎに、上蓋部123を保持部本体121に取り付け、腎臓S0を固定した〔図6(C)参照〕。なお、図6においては、押圧部22の記載を省略している。
実施例4において、対象物として腎臓、心臓、脾臓または肝臓を用いたことおよび対象物における標的部位に設置した圧力負荷部10aの条件を表3に示される条件としたことを除き、実施例4と同様の操作を行ない、ルシフェラーゼ活性を測定した。実施例5において、対象物の種類とルシフェラーゼ活性との関係を調べた結果を図21に示す。
実施例4において、対象物として腎臓の代わりに心臓を用いたことおよび送達装置1として、表4に示されたデバイス番号:5~7の送達装置それぞれを用いたことを除き、実施例4と同様の操作を行ない、ルシフェラーゼ活性を測定した。実施例6において、用いたデバイスの種類とルシフェラーゼ活性との関係を調べた結果を図22に示す。
麻酔下にあるマウスの腹腔を正中線に沿って切開し、腎臓を露出させた。その後、送達装置1の圧力負荷部10a(図2および図3参照)を腎臓における標的部位に設置した。なお、送達装置1として、表5に示されたデバイスを用いた。
対象物において、標的部位とは異なる部位に、薬剤、抗体、高分子化合物、金属ナノ粒子または量子ドットを溶媒に添加した混合物を注入し、標的部位に吸引圧または押圧力を負荷する。これにより、標的部位に、薬剤、抗体、高分子化合物、金属ナノ粒子または量子ドットを局所的に送達することができる。各物質に応じた検出方法により、標的部位における薬剤、抗体、高分子化合物、金属ナノ粒子または量子ドットの存在を確認することができる。
2 送達装置
10 圧力負荷部
10a 圧力負荷部
10b 圧力負荷部
11 吸引部
11a 吸引口
12 接続部
13 管体
21 保持部
22 押圧部
22a 押圧部本体
22b バルーン
22c 流路
22d 流体導入口
23 ポート
24 流路部
25 流体供給部
25a 保持部
26 流体貯留部
27 変換器
29 ニードル部
30 供給部
30a 貯留部
30b 送出部
40 制御部
121 保持部本体
121a 底部
121b 側部
121c 側部
121d 側部
122 管路側蓋部
123 上蓋部
131 突出部
132 端部
133 溝部
134 ストッパー
135 溝部
S0 組織
S1 標的部位
S2 管路(血管)
Claims (14)
- 生体内に存在する組織の標的部位の上流に位置する管路に導入対象物質を供給する供給部と、前記組織の標的部位に圧力を負荷する圧力負荷部とを備えた導入対象物質の送達装置の作動方法であって、
(1-1)供給部から導入対象物質を送出させ、前記管路に前記導入対象物質を供給する工程、および
(1-2)前記工程(1-1)と同時または前記工程(1-1)の後、圧力負荷部に前記組織の標的部位に対する圧力を発生させる工程
を含む導入対象物質の送達装置の作動方法。 - 前記導入対象物質が核酸またはペプチド化合物である請求項1に記載の方法。
- 前記圧力負荷部が、標的部位を吸引する吸引部を備えており、
前記工程(1-2)において、前記吸引部に前記標的部位を吸引させることにより、この標的部位に対する吸引圧を発生させる請求項1または2に記載の方法。 - 前記圧力負荷部が、標的部位を押圧する押圧部と、前記組織の位置を固定して保持する保持部とを備えており、
前記工程(1-2)において、前記押圧部に前記標的部位を押圧させることにより、この標的部位に対する押圧力を発生させる請求項1または2に記載の方法。 - 前記保持部が、生体内に埋め込み可能とされている請求項4に記載の方法。
- 非ヒト哺乳動物の組織の標的部位に導入対象物質を送達する導入対象物質の送達方法であって、
(2-1) 前記組織の上流に位置する管路に導入対象物質を供給する工程、および
(2-2) 前記工程(2-1)と同時または前記工程(2-1)の後、前記組織の標的部位に圧力を負荷して、当該標的部位に導入対象物質を送達する工程
を含む導入対象物質の送達方法。 - 前記導入対象物質が核酸またはペプチド化合物である請求項6に記載の方法。
- 前記工程(2-2)において、標的部位に対して吸引圧を負荷する請求項6または7に記載の方法。
- 前記工程(2-2)において、組織の位置を固定しながら、標的部位を押圧する請求項6または7に記載の方法。
- 前記組織が臓器の組織であり、
前記工程(2-2)において、前記臓器を収容可能なケースに臓器を収容して前記組織の位置を固定しながら、標的部位を押圧する請求項9に記載の方法。 - 生体由来の対象物の標的部位に導入対象物質を送達する導入対象物質の送達方法であって、
(3-1) 前記対象物において、標的部位とは異なる部位に導入対象物質を供給する工程、および
(3-2) 前記工程(3-1)と同時または前記工程(3-1)の後、前記対象物の標的部位に圧力を負荷して、当該標的部位に導入対象物質を送達する工程、
を含む対象物の標的部位への導入対象物質の送達方法。 - 前記導入対象物質が核酸またはペプチド化合物である請求項11に記載の方法。
- 前記工程(3-2)において、標的部位に対して吸引圧を負荷する請求項11または12に記載の方法。
- 前記工程(3-2)において、標的部位の位置を固定しながら、押圧する請求項11または12に記載の方法。
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JP2005040606A (ja) * | 2003-07-18 | 2005-02-17 | Cordis Corp | 肝臓潅流分離法 |
WO2005077317A1 (ja) * | 2004-02-13 | 2005-08-25 | Cms Co., Ltd. | 吸引マッサージ方法および吸引マッサージ装置 |
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JP2009057329A (ja) * | 2007-08-31 | 2009-03-19 | Aderans Holdings Co Ltd | 発毛促進システム |
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JPH11505112A (ja) * | 1995-05-04 | 1999-05-18 | ザ ボード オブ トラスティーズ オブ ザ リランド スタンフォード ジュニア ユニバーシティー | ヌクレオチドを細胞核内に送達するための装置および方法 |
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WO2005077317A1 (ja) * | 2004-02-13 | 2005-08-25 | Cms Co., Ltd. | 吸引マッサージ方法および吸引マッサージ装置 |
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