Injector and method for injecting solution using the same

Abstract

At least the following are provided. That is, provided is a technique for obtaining high physiological activity when a solution containing an in-vivo functional substance exhibiting physiological activity in a subject to be injected is injected into the subject to be injected. An injector for injecting a solution containing a functional substance in a living body into an object to be injected, the injector being provided with: a housing unit for housing the solution containing the functional substance in the living body; a pressurizing unit for pressurizing the solution containing the functional substance in the living body during operation; and an ejection part which can be inserted into the object to be injected, the ejection part having an inlet hole through which the solution containing the functional substance in the living body can flow in from the accommodating part when the solution containing the functional substance in the living body is pressurized by the pressurizing part, and an outlet hole through which the solution containing the functional substance in the living body can flow out from the accommodating part when the solution containing the functional substance in the living body is pressurized by the pressurizing part. The outlet hole can eject the inflowing solution containing the functional substance in the living body to the injection object, and the ejection speed of the solution containing the functional substance in the living body is greater than 83.3 [mu] L / s.

Description

Technical Field

[0001] The present disclosure relates to an injector and a method of injecting a solution using the injector. Background Art

[0002] As an injector for injecting a drug solution into an injection target, there are needle syringes that inject via an injection needle, needle-free syringes that inject without an injection needle, etc. In addition, in order to transport the drug solution to the injection target, a catheter equipped with an injection needle and a drive source, a multi-hole injection needle, etc. are also used.

[0003] Among them, in a needle syringe, the injection needle is inserted into the injection object, the plunger is pressed in, and the drug solution is usually slowly injected, thereby allowing the drug solution to diffuse in the injection object and exert its efficacy appropriately. In particular, when the drug solution is administered independently of the blood flow, a very small amount (for example, about 0.0033 μL / s) is slowly administered. In addition, in most cases, the administration is performed "slowly" based on the subjective "slowness" of the administering person.

[0004] On the other hand, needle-free syringes are used to instantly squeeze out liquid medicine from the tip of a nozzle using compressed gas, spring force, etc. as a driving force, and administer liquid medicine from the skin surface into a living body (Non-patent Document 1). In addition, a needle-free syringe that uses the combustion energy of a detonator as a driving force has been developed (Patent Document 1).

[0005] The needleless syringe that uses the combustion energy of the detonator as the ejection energy can deliver the medicine to the nucleus and cytosol by squeezing out the medicine liquid instantly. The needleless syringe does not necessarily need a viral vector or a lipid carrier to deliver the active ingredient to the injection object. Therefore, from the perspective of the drug delivery system (DDS), the usefulness of the needleless syringe has also attracted much attention, and is being studied in the delivery of various drugs such as low molecules, peptides, proteins, antibodies, etc. with anticancer effects. In particular, it is reported that in the field of vaccines and immunity, the needleless syringe can increase antibody titers and induce cellular immunity (non-patent literature 2). In addition, it is also reported that by using a needleless syringe to deliver plasmid DNA into the skin of experimental animals, the expression of genes is significantly enhanced compared with the delivery using a traditional needle syringe (non-patent literature 3).

[0006] The drug solution administered from the skin surface using a needleless syringe becomes a jet stream, penetrates the stratum corneum of the skin, and is delivered to the skin (Non-Patent Document 1). By adjusting the output, the drug solution can also be delivered to the subcutaneous or intramuscular area (Non-Patent Document 4).

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: Japanese Patent Application Publication No. 2012-61269

[0010] Non-patent literature

[0011] Non-patent document 1: Clin. Cosmet. Investig. Dermatol., 2018 May 1, 11: 231-238 Non-patent document 2: AAPS PharmSciTech., 2019 Dec 9, 21(1): 19

[0012] Non-patent literature 3: Journal of Pharmaceutical Sciences, 2019, 108: 2415-2420.

[0013] Non-patent literature 4: Gene, 2021, 788: 145664 Summary of the invention

[0014] Problems to be solved by the invention

[0015] When the inside of a living body is the injection target, a conventional syringe with a needle is generally used as an injector for injecting a drug solution, but there is room for improvement in terms of improving the drug efficacy.

[0016] The technical problem of the present disclosure is at least the following: That is, to provide a technology for obtaining high physiological activity when a solution containing a biological functional substance that exerts physiological activity in the injection target is injected into the injection target.

[0017] Solutions for solving problems

[0018] The present inventors have conducted intensive studies to solve the above-mentioned technical problems and have found that the above-mentioned technical problems can be solved by providing an injector that can be inserted into an injection portion of an injection target and can inject a solution at a predetermined high injection speed.

[0019] That is, the gist of the present disclosure is as follows.

[0020] [1] An injector for injecting a solution containing a biological functional substance into an injection object, the injector comprising: a container for containing the solution containing the biological functional substance; a pressurizing portion for pressurizing the solution containing the biological functional substance during operation; and an ejection portion capable of being inserted into the injection object, the ejection portion having an inlet hole and an outlet hole, and when the solution containing the biological functional substance is pressurized by the pressurizing portion, the inlet hole enables the solution containing the biological functional substance to flow into the container, and the outlet hole enables the solution containing the biological functional substance to be ejected into the injection object, wherein the ejection speed of the solution containing the biological functional substance is greater than 83.3 μL / s.

[0021] [2] The injector according to [1], wherein the injection speed of the solution containing the in vivo functional substance is 200 μL / s or more.

[0022] [3] The injector according to [1] or [2], wherein the inner diameter of the injection portion is 0.41 mm or less.

[0023] [4] The injector according to any one of [1] to [3], wherein the outer diameter of the injection portion is 0.72 mm or less.

[0024] [5] The injector according to any one of [1] to [4], wherein the ejection portion is an injection needle.

[0025] [6] The injector according to any one of [1] to [5], wherein the in vivo functional substance is one or more selected from the group consisting of nucleic acids, peptides, proteins, and low molecular weight compounds.

[0026] [7] An injector according to any one of [1] to [6], wherein the injection target is one or more selected from the group consisting of a joint cavity, spleen, eyeball, skin, muscle, bone, cartilage, bone marrow, ligament, brain, spinal cord, lung, liver, heart, kidney, pancreas, gallbladder, digestive tract, bladder, reproductive organ, lymph node, lymphatic network and blood vessel, a tumor in any of these, and mucous membrane and connective tissue attached to any of these.

[0027] [8] An injection method, comprising injecting the solution containing the in vivo functional substance into the injection target using the injector described in any one of [1] to [7].

[0028] Beneficial Effects

[0029] The present disclosure can achieve at least the following effects: that is, it can provide a technology for obtaining high physiological activity when a solution containing a biological functional substance that exerts physiological activity in the injection target is injected into the injection target. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 is an overall view of an injector according to one embodiment of the present disclosure.

[0031] Figure 2 Yes means Figure 1 A cross-sectional view of the vicinity of the top end portion of the injector is shown.

[0032] Figure 3 It is an overall view of a syringe according to a modified example.

[0033] Figure 4 yes Figure 3 A cross-sectional view of the injector is shown.

[0034] Figure 5 It is a graph showing the intra-articular Luc activity (total RFU) of the 2.5 mL Hamilton syringe (Syr) group with a 30G needle (n=4), the 30G-pNFI-25 / 40 (25 / 40Device) group (n=4), and the 30G-pNFI-35 / 40 (35 / 40Device) group (n=4). Mean + SD.

[0035] Figure 6 It is a graph showing the Luc activity (total RFU) in the joints of the group (n=4) of the administration rate of 0.25 μL / s (administration rate 0.25 μL / s) with a 25 mL Hamilton syringe with a 30G needle, the group (n=8) of the administration rate of 83.3 μL / s (administration rate 83.3 μL / s) with a 25 mL Hamilton syringe with a 30G needle, and the group (n=8) of 30G-pNFI-35 / 40. Mean + SD, Student t-test, *: P < 0.05.

[0036] Figure 7 It is a graph showing the Luc activity (total RFU) in the joints of the 30G-pNFI-35 / 40 (35 / 40) group, the 30G-pNFI-55 / 40 (55 / 40) group, and the 30G-pNFI-75 / 40 (75 / 40) group. Mean + SD.

[0037] Figure 8This is a graph showing the ejection speed of the solution when 30G-pNFI-25 / 40, 30G-pNFI-35 / 40, 30G-pNFI-55 / 40, 30G-pNFI-75 / 40, or pNFI-35 / 40 is used.

[0038] Fig. 9 This is a graph showing Luc activity (total RFU) in the spleen when 30G-pNFI-35 / 40 and an insulin syringe with a 30G injection needle (30G-Syr) were administered. Mean + SD.

[0039] Fig.10 This is a graph showing Luc activity (total RFU) in the spleen when 30G-pNFI-35 / 40 was administered using a Hamilton syringe with a 30G needle (30G-Syr / pump). Mean + SD.

[0040] Fig.11 This is a graph showing Luc activity (total RFU) in the spleen when 30G-pNFI-25 / 40, 30G-pNFI-35 / 40 and a Hamilton syringe with a 30G needle (30G-Syr / pump) were used for administration. Mean + SD.

[0041] Fig.12 This is a graph showing Luc activity (total RFU) in the testis when administered using 30G-pNFI-25 / 40, 30G-pNFI-35 / 40, and a Hamilton syringe with a 30G needle (30G-Syr / pump). Mean + SD.

[0042] Fig.13 This is a graph showing Luc activity (total RFU) in the caput epididymides when administered using 30G-pNFI-25 / 40, 30G-pNFI-35 / 40, and a Hamilton syringe with a 30G needle (30G-Syr / pump). Mean + SD.

[0043] Fig.14 This is a graph showing Luc activity (total RFU) in the cauda epididymis when administered using 30G-pNFI-25 / 40, 30G-pNFI-35 / 40, and a Hamilton syringe with a 30G needle (30G-Syr / pump). Mean + SD.

[0044] Fig.15 This is a graph showing the Luc activity (total RFU) in the liver when 30G-pNFI-25 / 40 and an insulin syringe with a 30G injection needle (30G-Syr) were administered. Mean + SD. DETAILED DESCRIPTION

[0045] Each configuration and their combination in each embodiment is an example, and the addition, omission, substitution and other changes of the configuration can be appropriately made without departing from the scope of the present disclosure. The present disclosure is not limited to the embodiments but only to the claims. In addition, each scheme disclosed in this specification can also be combined with any other feature disclosed in this specification.

[0046] Furthermore, a numerical range expressed using “to” means a range including the numerical values ​​described before and after “to” as the lower limit and the upper limit, and “A to B” means A or more and B or less.

[0047] One embodiment of the present disclosure is an injector, which is an injector for injecting a solution containing a biological functional substance into an injection object, the injector comprising: a container for containing the solution containing the biological functional substance; a pressurizing portion for pressurizing the solution containing the biological functional substance during operation; and an ejection portion capable of being inserted into the injection object, the ejection portion having an inlet hole and an outlet hole, when the solution containing the biological functional substance is pressurized by the pressurizing portion, the inlet hole enables the solution containing the biological functional substance to flow in from the container, and the outlet hole enables the inflow of the solution containing the biological functional substance to be ejected toward the injection object, and the ejection speed of the solution containing the biological functional substance is greater than 83.3 μL / s.

[0048] According to the injector of this embodiment, when a solution containing a biological functional substance that exhibits physiological activity in an injection target is injected into the injection target, high physiological activity is obtained.

[0049] <Solution containing biological functional substances>

[0050] The solution that can be injected by the injector of this embodiment includes a biological functional substance. The biological functional substance is not particularly limited as long as it exerts physiological activity in the injected object. The biological functional substance can be one or more. The biological functional substance can be a natural substance or an artificially synthesized biological functional substance.

[0051] The in vivo functional substance is preferably at least one selected from the group consisting of nucleic acids, peptides, proteins, and low molecular weight compounds.

[0052] Examples of the nucleic acid include DNA, RNA, and PNA. Furthermore, the nucleic acid may be a nucleic acid including a protein-encoding portion or a nucleic acid not including a protein-encoding portion (non-coding nucleic acid).

[0053] Examples of peptides and proteins include antigens (i.e., substances that produce antibodies against the peptides and proteins), antibodies, peptide vaccines, protein vaccines, peptide hormones, protein hormones, growth factors, cytokines, coagulation factors, serum albumin, digestive enzymes, anti-inflammatory peptides, anti-inflammatory proteins, and the like.

[0054] Low molecular weight compounds refer to compounds with a molecular weight of 2000 or less, but are not limited thereto, and include compounds that can be treated as low molecular weight compounds in the art. Preferred ranges of molecular weights of low molecular weight compounds include, for example, 50 or more and 100 or more. In addition, 2000 or less and 1000 or less can be listed. That is, 50 to 2000, 50 to 1000, and 100 to 2000 can be listed.

[0055] In the present disclosure, physiological activity refers to the effect on the specific physiological regulation function of an organism. The evaluation index of physiological activity can be appropriately set according to the purpose, and can be any kind of qualitative index or quantitative index, but preferably a quantitative index. For example, a specific mRNA amount, protein amount, cytokine amount, antibody titer, cell number of a specific cell type, etc. can be used as an index. These quantitative indices can be quantified by methods known in the art.

[0056] Particularly in the case where the functional substance in vivo is a nucleic acid comprising a portion of an encoded protein, the amount of the protein encoded by the nucleic acid can be quantified and used as an evaluation index for physiological activity. In addition, the activity of the protein can also be quantitatively evaluated. For example, as shown in the embodiments described later, when the protein is a luciferase, physiological activity can be evaluated by measuring the intensity of bioluminescence.

[0057] When the functional substance in the organism is a nucleic acid, the nucleic acid can be integrated into a viral vector or carried on a lipid nanoparticle and included in the solution, but according to the injector of this embodiment, high physiological activity can be obtained even without using a viral vector or lipid nanoparticle. Without using a viral vector or lipid nanoparticle, the possibility of inducing side effects such as allergic reactions in the injected object can be reduced.

[0058] The content of the biofunctional substance relative to the total amount of the solution can be appropriately set based on the type of the biofunctional substance, the injection target, the physiological activity of the biofunctional substance in the injection target, and the like.

[0059] The solution may contain, in addition to the in vivo functional substance, common additives such as a buffer, an isotonicity regulator, a pH adjuster, an antioxidant, a thickener, a stabilizer, a wetting agent, an emulsifier, and a binder, as required.

[0060] As the buffer, for example, a buffer using phosphate (e.g., phosphate buffer, phosphate buffered saline (PBS) (which may be PBS(+) or PBS(-)), Dulbecco's phosphate buffered saline (D-PBS), citric acid / phosphate buffer, citric acid / phosphate buffered saline, etc.), citrate buffer, trishydroxymethylaminomethane-HCl buffer (Tris-HCl buffer), acetate buffer, GOOD buffer (e.g., HEPES-NaOH buffer, etc.), amino acid buffer (e.g., glycine-HCl buffer, glycine-NaOH buffer, glycylglycine-NaOH buffer, glycylglycine-KOH buffer, etc.), imidazole buffer, etc. can be used.

[0061] Among them, from the viewpoint of versatility, it is preferred to use a phosphate buffer.

[0062] Examples of the isotonicity adjusting agent include ionic isotonicity adjusting agents and nonionic isotonicity adjusting agents.

[0063] Examples of the ion isotonicity adjusting agent include salts such as sodium chloride, potassium chloride, calcium chloride, and magnesium chloride.

[0064] Examples of the nonionic isotonicity adjusting agent include glycerin, propylene glycol, polyethylene glycol, glucose, sorbitol, mannitol, trehalose, maltose, and sucrose.

[0065] Among them, sodium chloride is preferred from the viewpoint of versatility.

[0066] Examples of the pH adjuster include hydrochloric acid, phosphoric acid, citric acid, acetic acid, sodium hydroxide, potassium hydroxide, sodium carbonate, and sodium hydrogen carbonate.

[0067] Examples of the antioxidant include ascorbic acid, sodium sulfite, butylhydroxyanisole, butylhydroxytoluene, propyl gallate, and tocopherol.

[0068] Examples of the thickener include alginic acid, polyethylene glycol, hydroxypropylmethylcellulose, and sodium carboxymethylcellulose.

[0069] The content of the in vivo functional substance in the solution is not particularly limited and can be appropriately adjusted by a person skilled in the art within the range in which the in vivo functional substance can exert physiological activity. For example, it can be listed as: 0.0001 mg / mL or more, 0.001 mg / mL or more, 0.01 mg / mL or more, etc. In addition, it can be listed as: 1000 mg / mL or less, 100 mg / mL or less, etc. That is, it can be listed as: 0.0001 to 1000 mg / mL, 0.001 to 100 mg / mL, 0.01 to 100 mg / mL, etc.

[0070] The pH of the solution is not particularly limited as long as the in vivo functional substance exhibits physiological activity and stably exists in the injection target when the solution is injected into the injection target, and does not cause adverse effects such as damaging the injection target.

[0071] <Injector>

[0072] As described above, the injector of this embodiment comprises: a containing portion for containing a solution containing a functional substance in a living body; a pressurizing portion for pressurizing the solution; and an ejection portion that can be inserted into an injection target of the solution and injects the solution into the injection target by ejecting the solution from an outlet hole of the ejection portion.

[0073] In the injector of this embodiment, the energy imparted by the pressurizing part for pressurizing the solution containing the functional substance in the organism can be applied by the energy imparted by the known pressurizing technology. As an example of the energy imparted, it can be chemically generated energy, such as combustion energy generated by the oxidation reaction of gunpowder, explosives, etc. In addition, as another method, the energy used for the pressurization can also be generated by electricity, as an example, it can be the energy of the piezoelectric element or electromagnetic driver driven by the electric power introduced. Furthermore, as another method, the energy used for the pressurization can also be physically generated, as an example, it can be the elastic energy from the elastic body, the internal energy of the compressed object such as compressed gas. That is, the energy used for the pressurization can be any energy as long as it can eject the solution in the injector. In addition, the energy used for the pressurization can also be a composite energy that appropriately combines these combustion energies, energy from electricity, elastic energy and other internal energies.

[0074] According to the above, the pressurizing part may be, for example, a pressurizing part that uses the pressure generated by the combustion of gunpowder ignited by the ignition device to pressurize, or a pressurizing part that uses the pressure generated when the compressed gas is released to pressurize. In addition, the pressurizing part may be a pressurizing part that uses the applied force of a compression spring to pressurize, or a pressurizing part that uses electromagnetic force to pressurize, such as a pressurizing part that uses a linear electromagnetic drive to pressurize. The pressurizing part is preferably a method that uses at least the pressure generated by the combustion of gunpowder ignited by the ignition device, and further, it may be used in combination with any of the other pressurizing methods mentioned above.

[0075] In the case where the pressurizing part utilizes the pressure generated by the combustion of gunpowder, the gunpowder may be, for example, any of the following gunpowders, or a combination of multiple thereof: gunpowder containing zirconium and potassium perchlorate (ZPP), gunpowder containing titanium hydride and potassium perchlorate (THPP), gunpowder containing titanium and potassium perchlorate (TiPP), gunpowder containing aluminum and potassium perchlorate (APP), gunpowder containing aluminum and bismuth oxide (ABO), gunpowder containing aluminum and molybdenum oxide (AMO), gunpowder containing aluminum and copper oxide (ACO), gunpowder containing aluminum and iron oxide (AFO). As a feature of these gunpowders, even if their combustion products are gaseous at high temperature, they do not contain gas components at room temperature, so the combustion products are condensed immediately after ignition.

[0076] In addition, when the pressurizing part uses the energy generated by the combustion of the gas generating agent as the injection energy, the gas generating agent may be a single-base smokeless powder (GG) or various gas generating agents used in gas generators for air bags and gas generators for seat belt pretensioners.

[0077] The injector of the present embodiment may be configured as a needle-equipped syringe having an injector body and an injection needle, wherein the injector body has a housing and a pressurizing portion, and the injection needle is mounted on the injector body as an ejection portion. In addition, with respect to the injector of the present embodiment, the housing and ejection portion of the injector body may be integrally formed. In addition, the injector body may be configured as a needle-free syringe that can be injected alone without an injection needle, for example. However, the injector body of the present invention is not limited to a needle-free syringe. The injector body may be, for example, a syringe pump. In addition, the ejection portion of the present invention is not limited to an injection needle. For example, in the case where the distance from the injector body to the injection object is large, a catheter or a porous injection needle may be used as the ejection portion so that a solution containing a functional substance in a biological body can be guided from the housing of the injector body to the injection object.

[0078] Here, from the viewpoint of suppressing tissue damage of the injection object, the ejection portion inserted into the injection object is preferably thinner. Specifically, the inner diameter of the ejection portion is preferably less than 0.41 mm, and the outer diameter of the ejection portion is preferably less than 0.72 mm. However, the ranges of the inner diameter and outer diameter of the ejection portion disclosed in the present invention are not limited to the above. In addition, the length of the ejection portion can be appropriately set according to the injection object. For example, in the case where the ejection portion is constituted by an injection needle, the length of the injection needle can use a value selected from the range of more than 1 mm and less than 180 mm. If the length of the injection needle is more than 1 mm, the injection needle can reach the skin when the injection needle is pierced vertically on the skin. In addition, even if the deep internal organs of a person are assumed to be the injection object, as long as the length of the injection needle is 180 mm, the injection needle can fully reach the deep internal organs. In addition, the material of the ejection portion is not particularly limited, and metal materials and resin materials can be used.

[0079] In the injector of this embodiment, the solution containing the in vivo functional substance may be contained in the containing section from the beginning, or the solution may be contained in the containing section by sucking the solution from the outside of the injector through the outlet hole of the ejection section.

[0080] [syringe]

[0081] Hereinafter, referring to the accompanying drawings, an example of an injector according to the present embodiment will be described. Figure 1 , Figure 2 The syringe 100 shown in the figure is described. It should be noted that the configuration of the following embodiment is an example, and the technology disclosed in the present invention is not limited to the configuration of the embodiment. It should be noted that in the following description, "top end side" and "base end side" are used as terms to indicate the relative positional relationship in the longitudinal direction of the syringe 100. The "top end side" refers to the side of the syringe 100 in the longitudinal direction. Figure 1 Regarding the outlet hole 4 b side shown in the figure, the “base end side” refers to the side opposite to the outlet hole 4 b side in the longitudinal direction of the syringe 100 .

[0082] Figure 1 : is an overall view of the syringe 100 of this embodiment. Figure 1 , the figure shows a cross section along the long dimension direction of the syringe 100. In addition, Figure 2 2 is a cross-sectional view showing the vicinity of the top end portion of the syringe 100. Figure 1 As shown, the syringe 100 includes a jet syringe 1 as an injector body, a housing 2 for accommodating the jet syringe 1 , an injection needle 4 as an injection portion, and a fixing jig 5 for fixing the injection needle 4 to the jet syringe 1 .

[0083] [Jet Injector]

[0084] like Figure 1As shown, the jet injector 1 includes a container 11, a container holder 12, a driver 13 and a housing 14, which are an assembly formed by assembling them in one piece. An ejection hole shown by the reference numeral 11c is formed at the top of the jet injector 1. The jet injector 1 uses the combustion energy of gunpowder to pressurize a solution (hereinafter sometimes simply referred to as a "solution") containing a functional substance in a living body, thereby ejecting the solution from the ejection hole 11c. The jet injector 1 of this embodiment is configured as a needle-free injector that can be injected alone without an injection needle. However, the jet injector 1 may not be a needle-free injector.

[0085] [case]

[0086] like Figure 1 As shown in FIG. 1 , the housing 14 is a cylindrical member that accommodates the driver 13. An ignition device 131 (described later) of the driver 13 is embedded in the base end side of the housing 14, and the container holder 12 is embedded in the top end side of the housing 14. Figure 2 As shown, a screw portion 14a for coupling the housing 14 to the container holder 12 is formed on the inner peripheral surface of the front end side of the housing 14. The container holder 12 and the driver 13 are connected via the housing 14.

[0087] [container]

[0088] like Figure 2 As shown, the container 11 has a main body 111 and a nozzle 112 having a smaller diameter than the main body 111, and is formed in a cylindrical shape. In addition, a storage space 11a as a space capable of storing a solution containing a functional substance in a living body, and a flow path 11b connected to the storage space 11a and opened at the top end side are formed inside the container 11. In more detail, the storage space 11a is formed inside the main body 111, and the flow path 11b is formed inside the nozzle 112. The storage space 11a is an example of a storage portion of the present disclosure. An opening portion of the flow path 11b is formed on the top end surface of the nozzle 112, and the opening portion serves as an ejection hole 11c of the jet syringe 1. That is, the jet syringe 1 ejects the solution from the ejection hole 11c formed on the top end surface of the nozzle 112. As shown in FIG. Figure 2 As shown, the inner diameter of the flow path 11b of the container 11 is smaller than the inner diameter of the storage space 11a. With such a structure, the solution pressurized to a high pressure is ejected to the outside from the ejection hole 11c of the flow path 11b. In addition, a threaded portion 111a for connecting the container 11 to the container holder 12 is formed on the outer peripheral surface of the main body 111.

[0089] The material of the container 11 is not particularly limited, and for example, the container 11 may be formed of a resin material. As the resin material forming the container 11, for example, known nylon 6-12, polyarylate, polycarbonate, polybutylene terephthalate, polyphenylene sulfide, or liquid crystal polymer may be used.

[0090] [Container support]

[0091] like Figure 2 As shown, the container holder 12 is formed in a cylindrical shape and holds the container 11 embedded in the container holder 12. A threaded portion 12a is formed on the inner peripheral surface of the base end side of the container holder 12. The container 11 is combined with the container holder 12 by screwing the threaded portion 111a of the container 11 with the threaded portion 12a of the container holder 12. In addition, a threaded portion 12b is formed on the outer peripheral surface of the base end side of the container holder 12. The container holder 12 is combined with the shell 14 by screwing the threaded portion 12b of the container holder 12 with the threaded portion 14a of the shell 14. In addition, a threaded portion 12c for combining the container holder 12 with the fixing fixture 5 is formed on the outer peripheral surface of the top end side of the container holder 12.

[0092] The material of the container holder 12 is not particularly limited, and for example, the container holder 12 may be formed of a metal material. Examples of the metal material forming the container holder 12 include stainless steel, copper, aluminum, iron, titanium, and titanium alloys.

[0093] [Drive]

[0094] Figure 1 The actuator 13 shown is configured to pressurize the solution when it is in operation. The actuator 13 is an example of a "pressurizing unit" of the present disclosure. Figure 1 As shown, the driver 13 includes an ignition device 131 , a piston 132 , and a plunger 133 accommodated in the housing 14 .

[0095] The ignition device 131 is disposed at the base end of the housing 14, the plunger 133 is disposed at the top end of the housing 14, and the piston 132 is adjacent to the plunger 133 and disposed between the ignition device 131 and the plunger 133. In the internal space of the housing 14, a combustion chamber 15 is formed between the ignition device 131 and the piston 132.

[0096] [Ignition device]

[0097] The ignition device 131 includes an igniter 1311 and a retaining member 1312. The igniter 1311 serves as a driving source of the driver 13, and generates energy for causing the syringe 100 to pressurize and eject the solution. The igniter 1311 is configured as an electric igniter that releases combustion products by burning the explosive contained therein. The retaining member 1312 is formed by injection molding of a resin. The injection molding can use a known method. In addition, the retaining member 1312 can use the same resin material as the container 11. The ignition device 131 is configured as an igniter assembly in which the igniter 1311 is fixed to the base end of the housing 14 via the retaining member 1312, and is embedded in the housing 14 in a manner that blocks the base end of the housing 14. In addition, the ignition device 131 is arranged in the housing 14 in a manner that the igniter 1311 and the base end surface of the piston 132 are opposite to each other, so as to transfer the combustion energy of the explosive of the igniter 1311 and the combustion energy of the gas generator 10 described later to the base end surface (end surface on the base end side) of the piston 132.

[0098] [Detonator]

[0099] Here, as the explosive for the igniter 1311, examples include gunpowder containing zirconium and potassium perchlorate (ZPP), gunpowder containing titanium hydride and potassium perchlorate (THPP), gunpowder containing titanium and potassium perchlorate (TiPP), gunpowder containing aluminum and potassium perchlorate (APP), gunpowder containing aluminum and bismuth oxide (ABO), gunpowder containing aluminum and molybdenum oxide (AMO), gunpowder containing aluminum and copper oxide (ACO), gunpowder containing aluminum and iron oxide (AFO), or gunpowder composed of a combination of multiple of these gunpowders. These gunpowders generate high-temperature and high-pressure plasma when they burn immediately after ignition, but if the combustion products are condensed at room temperature, they do not contain gas components, and therefore show the characteristic of a sharp drop in the generated pressure. It should be noted that as long as an appropriate solution can be injected, gunpowders other than these can also be used as the explosive.

[0100] [Gas generating agent]

[0101] In addition, in the jet injector 1, in order to adjust the pressure change applied to the solution via the piston 132, a gas generator 10 is arranged in the combustion chamber 15, and the gas generator 10 generates gas by burning the combustion product of the explosive released from the igniter 1311. That is, the jet injector 1 is configured to pressurize the solution by using the combustion energy of the gas generator 10 in addition to the combustion energy of the explosive of the igniter 1311. The arrangement position of the gas generator 10 is a position where it can be exposed to the combustion product from the igniter 1311. In addition, as another method, the gas generator 10 can also be arranged in the igniter 1311 as disclosed in International Publication No. 01-031282, Japanese Patent Publication No. 2003-25950, etc. As an example of the gas generator, a single-base smokeless powder (GG) composed of 98% by mass of nitrocellulose, 0.8% by mass of diphenylamine, and 1.2% by mass of potassium sulfate can be cited. In addition, various gas generators used in gas generators for airbags and seat belt pretensioners can also be used. By adjusting the size, size, shape, and especially the surface shape of the gas generator when it is arranged in the combustion chamber 15, the combustion end time of the gas generator can be changed, thereby adjusting the pressure shift applied to the solution to cause the desired shift in the injection pressure. In addition, the jet injector 1 may not include the gas generator 10 and only use the combustion energy of the explosive of the igniter 1311 to pressurize the solution. In the present disclosure, the pressurizing unit also includes a gas generator used as needed.

[0102] [piston]

[0103] The piston 132 is arranged on the top side of the housing 14 so as to be pressurized by the operation of the igniter 1311 and to be able to slide inside the housing 14. The piston 132 is made of metal, and in order to improve the closeness with the sliding surface (i.e., the inner peripheral surface of the housing 14) on which the piston 132 will slide, an O-ring or the like may be arranged on a part of the piston 132. As another method, the piston 132 may also be made of resin, in which case metal may also be used in the portion requiring heat resistance and pressure resistance.

[0104] [Plunger]

[0105] The plunger 133 is a member that pressurizes the solution contained in the storage space 11a by receiving the combustion energy of the detonator of the igniter 1311 and the combustion energy of the gas generating agent through the piston 132. The plunger 133 is contained in the housing 14 and is arranged between the piston 132 and the nozzle portion 112 of the container 11. The plunger 133 is formed in a rod shape, and its base end is engaged with the top end of the piston 132. In addition, the top end of the plunger 133 is inserted into the main body 111 of the container 11, and the storage space 11a is defined by the plunger 133 and the main body 111. The plunger 133 can slide inside the main body 111, and the solution contained in the storage space 11a is pressurized and ejected from the ejection hole 11c through the flow path 11b by sliding the plunger 133. Therefore, the plunger 133 is formed of a material that can slide smoothly relative to the main body 111 of the container 11 and can prevent the solution from leaking from the plunger 133 side.

[0106] As a specific material of the plunger 133, for example, butyl rubber or silicone rubber can be used. In addition, in order to ensure / adjust the sliding property between the plunger 133 and the main body 111 of the container 11, the outer peripheral surface of the plunger 133 and the inner peripheral surface of the main body 111 can also be coated / surface-processed with various substances.

[0107] Here, the contour of the top end of the plunger 133 is roughly the same as the contour of the top end of the storage space 11a. Therefore, when the plunger 133 slides and reaches the deepest position in the main body 111 during the injection of the solution, the storage space 11a formed between the plunger 133 and the nozzle 112 can be reduced as much as possible, thereby preventing the solution from remaining in the storage space 11a and being wasted.

[0108] [shell]

[0109] Figure 1 The housing 2 shown is a member for accommodating the jet injector 1, and functions as a grip portion for the user to grasp in order to use the injector 100. Figure 1 As shown in FIG. 1 , a battery 3 for supplying a driving current to a driver 13 (more specifically, an igniter 1311) of the jet injector 1 is provided inside the housing 2. Figure 1As shown, a plurality of switches 21 for operating the jet injector 1 to eject the solution are provided on the outer surface of the housing 2. In addition, a socket (not shown) connected to the igniter 1311 of the jet injector 1 is provided on the inner surface of the housing 2. The power supply from the battery 3 to the jet injector 1 is performed by the user pressing the button 21, and is performed between the electrode on the side of the housing 2 and the electrode on the side of the igniter 1311 of the jet injector 1 via wiring. In addition, a control unit such as a microcomputer (not shown) is provided inside the housing 2. The control unit controls the supply of ignition current to the igniter 1311 of the jet injector 1 based on the signal from each switch, thereby controlling the operation of the jet injector 1.

[0110] It should be noted that, as described above, in the present embodiment, the power for operating the igniter 1311 is supplied from a battery built into the housing 2 , but instead of this, the power may be supplied from the outside via a power cable.

[0111] [Injection needle]

[0112] like Figure 1 As shown, the injection needle 4 is mounted on the nozzle portion 112 of the jet injector 1, and is configured to include a base portion 41 and a needle tube 42. The injection needle 4 is an example of the "injection portion" of the present disclosure, and is configured to allow the solution pressurized by the driver 13 of the jet injector 1 to flow in and to eject the inflowing solution toward the injection object.

[0113] The solution pressurized by the driver 13 flows into the base 41. Figure 1 As shown in FIG. 4 , the base 41 is formed into a cylindrical shape. Figure 2 As shown, the base 41 includes a cylindrical base body 411 and a flange portion 412 formed at the base end of the base body 411 and extending radially outward. The nozzle portion 112 of the jet injector 1 is pressed into the opening on the base end side of the base body 411. Therefore, the opening on the base end side of the base body 411 is configured as an inlet hole 4a, which allows a solution containing a functional substance in the body to flow from the storage space 11a of the container 11 into the injection needle 4.

[0114] The base 41 can be formed of a resin material, for example. As the resin material forming the base 41, for example, a known synthetic resin such as polycarbonate, polypropylene, polyethylene, etc. can be used. In addition, the base 41 can also be made of metal. As the metal material forming the base 41, for example, stainless steel, aluminum, aluminum alloy, titanium, titanium alloy, etc. can be exemplified. The material of the base 41 is not particularly limited, but if the pressure loss of the solution flowing into the injection needle 4 is considered, it is preferable to use a material with high rigidity such as stainless steel for the base 41.

[0115] The needle tube 42 is inserted into the injection object, and the solution containing the functional substance in the living body is ejected to the injection object. The base end of the needle tube 42 is connected to the top end of the base 41, and the internal space of the base 41 is connected to the internal space of the needle tube 42. Therefore, the opening on the top side of the needle tube 42 is configured as an outlet hole 4b, and the outlet hole 4b can eject the solution containing the functional substance in the living body that flows into the injection needle 4 from the inlet hole 4a to the injection object. In addition, in order to insert the needle tube 42 into the injection object, a sharp needle tip for piercing the skin is formed at the top end of the needle tube 42.

[0116] Here, from the viewpoint of suppressing tissue damage of the injection target, the injection needle 4 (more specifically, the needle tube 42 inserted into the injection target) is preferably thinner. Figure 2 As shown in the enlarged view A1 of , the inner diameter of the needle tube 42 is set to d1, and the outer diameter of the needle tube 42 is set to d2. At this time, the inner diameter d1 is preferably less than 0.41 mm, and the outer diameter d2 is preferably less than 0.72 mm. However, the range of the inner diameter and outer diameter of the needle tube 42 is not limited to the above range. In addition, the length of the needle tube 42 can be appropriately set according to the injection object, for example, a value selected from the range of more than 1 mm and less than 180 mm can be used.

[0117] The material of the needle tube 42 is not particularly limited, and for example, stainless steel can be cited. In addition, as a metal material other than stainless steel, aluminum, aluminum alloy, titanium, titanium alloy, etc. can be used. In addition, the needle tube 42 can also be formed of a resin material.

[0118] As described above, since the internal space of the base 41 is connected to the internal space of the needle tube 42, a flow path 4c from the inlet hole 4a to the outlet hole 4b is formed in the injection needle 4 through the internal space of the base 41 and the internal space of the needle tube 42. The solution flowing into the injection needle 4 from the inlet hole 4a flows through the flow path 4c to the outlet hole 4b, and is ejected from the outlet hole 4b toward the injection object.

[0119] A sealant may be used between the nozzle portion 112 of the container 11 and the base portion 41 of the injection needle 4 to improve the liquid tightness. Examples of the sealant include O-rings, seals, sealing tapes, and liquid sealants.

[0120] [Fixed fixture]

[0121] The fixing jig 5 is a member for fixing the injection needle 4 to the nozzle portion 112 of the jet injector 1. The fixing jig 5 includes a cylindrical peripheral wall portion 51 and a cover wall portion 52 that blocks the top end of the peripheral wall portion 51. The container holder 12 is embedded in the peripheral wall portion 51. In addition, a threaded portion 51a is formed on the inner peripheral surface of the peripheral wall portion 51, and the container holder 12 is coupled to the fixing jig 5 by screwing the threaded portion 12c of the container holder 12 and the threaded portion 51a of the fixing jig 5. A through hole 52a is formed in the cover wall portion 52, which passes through the cover wall portion 52 from the base end side to the top end side. The base body 411 of the injection needle 4 is embedded in the through hole 52a. At this time, the cover wall portion 52 presses the flange portion 412 of the injection needle 4 from the top end side, thereby preventing the injection needle 4 from falling off from the nozzle portion 112 of the jet injector 1. In this way, the injection needle 4 is fixed to the nozzle portion 112 by the fixing jig 5.

[0122] The material of the fixing jig 5 is not particularly limited, and similarly to the container holder 12, the fixing jig 5 may be made of metal materials such as stainless steel, copper, aluminum, iron, titanium, and titanium alloy. Alternatively, the fixing jig 5 may be made of resin materials such as polycarbonate, polypropylene, and polyethylene.

[0123] [Injector Action]

[0124] The injection of a solution into an injection object by using the syringe 100 is performed by operating the syringe 100 while the needle tube 42 of the injection needle 4 is inserted into the injection object. The operation of the syringe 100 is described below. It should be noted that the solution containing the biological functional substance can be contained in the storage space 11a of the container 11 from the beginning, or the solution can be sucked from the outside of the syringe 100 through the outlet hole 4b of the injection needle 4 to be contained in the storage space 11a. In the case where the storage part and the injection part are integrally formed, by containing the solution containing the biological functional substance in the storage space 11a of the container 11 from the beginning, it is possible to reduce leakage of the drug solution and dead volume.

[0125] When the driver 13 of the jet injector 1 is operated by the user's operation of the switch 21, a driving current is supplied to the driver 13 (more specifically, the igniter 1311), and the driver 13 is operated, the combustion product of the explosive is released from the igniter 1311 to the combustion chamber 15. As a result, the gas generating agent 10 disposed in the combustion chamber 15 is burned by the combustion product of the explosive, and gas is generated in the combustion chamber 15. As described above, the base end surface of the piston 132 is exposed to the combustion chamber 15. Therefore, when the driver 13 is operated, the piston 132, which receives the combustion energy (pressure) of the explosive and the gas generating agent at the base end surface, slides toward the top end side of the housing 14. As a result, the plunger 133 is pressed into the top end side of the storage space 11a by the piston 132, and the solution in the storage space 11a is pressurized. As a result, the solution passes through the flow path 11b from the storage space 11a and is ejected from the ejection hole 11c formed in the nozzle portion 112.

[0126] The solution ejected from the ejection hole 11c of the nozzle 112 flows into the injection needle 4 through the inlet hole 4a of the injection needle 4 attached to the nozzle 112. The solution flows through the flow path 4c and is ejected from the outlet hole 4b to the injection target. As described above, the operation of the syringe 100 is completed.

[0127] [Modifications]

[0128] Figure 3 FIG. 1 is an overall view of a syringe 100A according to a modified example of the present embodiment. Figure 3 , the appearance of the syringe 100A is shown. Figure 4 is a cross-sectional view of the syringe 100A. Figure 3 As shown in FIG. 1 , a power cable 6 for supplying a driving current to a driver 13 of the jet injector 1 is provided in the housing 2 of the injector 100A. Figure 4 As shown, the driver 13 of the jet injector 1 of the modified example has a cylindrical body 134 that accommodates an ignition device 131 and a piston 132. The ignition device 131 is arranged on the base end side of the body 134, and the piston 132 is arranged on the top end side. In the internal space of the body 134, a combustion chamber 15 is formed between the ignition device 131 and the piston 132. The injector of the present disclosure can be implemented in a structure like the injector 100A.

[0129] [Solution ejection speed]

[0130] The injector of this embodiment can eject the solution containing the functional substance in vivo at an ejection speed greater than 83.3 μL / s. The ejection speed is preferably 200 μL / s or more, more preferably 250 μL / s or more, further preferably 300 μL / s or more, further preferably 350 μL / s or more, further preferably 400 μL / s or more, and most preferably 500 μL / s or more. In addition, the upper limit is not particularly limited, and 5000 μL / s or less can be cited. That is, the preferred range of the injection speed includes greater than 83.3 μL / s and less than 5000 μL / s, greater than 200 μL / s and less than 5000 μL / s, greater than 250 μL / s and less than 5000 μL / s, greater than 300 μL / s and less than 5000 μL / s, greater than 350 μL / s and less than 5000 μL / s, greater than 400 μL / s and less than 5000 μL / s, and greater than 500 μL / s and less than 5000 μL / s.

[0131] If the injection speed is greater than 83.3 μL / s, high physiological activity can be obtained when the solution is injected into the injection object. The detailed mechanism is not clear, but it is speculated that if shear stress is applied to the surrounding tissues / cells, endocytosis is hyperactive and the delivery of functional substances in the body to the cells is improved. Based on the above mechanism, it is believed that the higher the injection speed within the above range, the more preferred.

[0132] When the ejection speed is equal to or lower than the above upper limit, damage to cells and tissues can be suppressed.

[0133] In the present disclosure, the injection speed is the speed of the solution when it is injected from the injection hole, and can be calculated, for example, by dividing the injected volume by the time required from the start to the end of the injection. The time required from the start to the end of the injection can be measured, for example, by photographing the situation of the solution being injected from the injection hole with a photographing device such as a high-speed camera.

[0134] The injection speed can be adjusted to be within the above range by adjusting the injection energy according to the shape and material of the injection portion and the volume and viscosity of the solution.

[0135] <Injection object>

[0136] The biological species of the injection target that the injector of this embodiment can inject are not particularly limited, and mammals can be listed, for example, humans, mice, rats, guinea pigs, hamsters, cattle, goats, sheep, pigs, monkeys, dogs, cats, etc. In addition, mammals other than humans can also be listed.

[0137] The injection object can be set to the inside of the organism, but is not limited to this. The inside of the organism refers to a part that is not the outer surface of the organism, which can be the surface part of an organ inside the organism, the inside of an organ, or a space such as a body cavity. Specifically, it is preferably selected from the group consisting of a joint cavity, spleen, eyeball, skin, muscle, bone, cartilage, bone marrow, ligament, brain, spinal cord, lung, liver, heart, kidney, pancreas, gallbladder, digestive tract, bladder, reproductive organ, lymph node, lymphatic network and blood vessel, a tumor at any of these, and a mucous membrane and connective tissue attached to any of these. As reproductive organs, testicles, ovaries, etc. can be listed. Among them, it is preferably selected from the group consisting of a joint cavity, spleen, testicles and liver.

[0138] When the injection target is one or more selected from the group consisting of spleen, testis and liver, the lower limit of the injection speed of the injector of this embodiment can be 1 μL / s or more, and can be 10 μL / s or more. By injecting the solution into the injection target at the injection speed within the above range, sufficient physiological activity can be obtained.

[0139] The injection amount of the solution into the injection target can be appropriately set based on the content and type of the in vivo functional substance, the injection target, the physiological activity of the in vivo functional substance in the injection target injected with the in vivo functional substance, and the like.

[0140] The injection amount of the solution into the injection object can be, for example, 10 μL or more, 50 μL or more, or 100 μL or more. In addition, it can be 50 mL or less, 10 mL or less, or 5 mL or less. That is, it can be 10 μL to 50 mL, 50 μL to 10 mL, or 100 μL to 5 mL.

[0141] The time required for injecting the solution into the injection object is not particularly limited as long as it is the time for injecting the solution of the injection amount at the injection speed, and examples thereof include: 0.005 seconds or more, 0.01 seconds or more, or 0.1 seconds or more. In addition, examples thereof include: 500 seconds or less, 100 seconds or less, or 10 seconds or less. That is, examples thereof include: 0.005 to 500 seconds, 0.01 to 100 seconds, 0.1 to 10 seconds, etc.

[0142] Example

[0143] Hereinafter, the present disclosure will be specifically described with reference to examples. However, the present disclosure is not limited to the schemes of the following examples.

[0144] (animal)

[0145] SD rats (8 weeks old, male, SLC) or ICR mice (8 weeks old, male, SLC) were used.

[0146] All surgical procedures and syringe or device administration were performed using a gas anesthesia device (Nacrobit-E, Natsume Seisakusho) under isoflurane (Pfizer) anesthesia maintenance.

[0147] All animal experiments were approved by the Animal Experiment Review Committee of the Natural Products Research Institute Co., Ltd. (Kawasaki City) or the National Institute of Animal Reproduction (Kasumigaura City), where the experiments were conducted.

[0148] (Injection of solution)

[0149] The administration to the knee joint of the rat was performed as follows: the left and right knee joints of the rat were shaved, and under isoflurane anesthesia, the needle tip was inserted to a depth of 5 mm with the target being 5 mm below the median patella and the upper end of the tibia, and the plasmid pGL4 (hereinafter sometimes referred to as pLuc) for luciferase (Luc) expression was administered. pGL4 was diluted to 1 mg / mL in advance using PBS.

[0150] When administering to the spleen, testicles, or liver of mice, the left flank, the periphery of the scrotum, or the central part of the abdomen was shaved under triple anesthesia, and the administration organ was exposed by laparotomy under anesthesia, and pLuc was injected. After confirming that there was no bleeding at the administration site, the muscles and skin at the incision site were ligated and sutured, and the abdomen was closed.

[0151] (Injector)

[0152] As the injector, an insulin syringe with a 30G injection needle (BD Company), a Hamilton syringe (Luer lock type, syringe capacity 50-50000 μL, Hamilton Company) equipped with a 30G injection needle (disposable needle, NO.30 Dentronics injection needle, 0.3×12 mm, Dentronics Company), or a jet injector (pNFI, refer to Figure 1 ). In manual administration with a 30G-Hamilton syringe, administration is performed by slowly pressing the plunger for about 1 second (about 30 μL / s). In administration with a syringe pump of a 30G-Hamilton syringe, the administration speed program of the syringe pump is used (0.25 to 83.3 μL / s). On the other hand, in the case of a jet syringe, ZPP is used as a detonator, and GG (containing 98% by mass of nitrocellulose, 0.8% by mass of diphenylamine, and 1.2% by mass of potassium sulfate) is used as a gas generator, and these are used to administer the solution. The ejection speed of the jet syringe is measured by the test described below. In addition, as Figure 2As shown, the injection needle 4 is fixed to the tip of the nozzle portion 112 of the jet injector by a metal fixing clamp 5. Hereinafter, for example, a jet injector equipped with 35 mg of ZPP and 40 mg of GG is referred to as "pNFI-35 / 40", and a jet injector further equipped with a 30G injection needle is referred to as "30G-pNFI-35 / 40".

[0153] (Collection of Organ Specimens)

[0154] Dissection and organ specimen collection were performed under isoflurane anesthesia after confirming death by exsanguination of the carotid artery after cervical dislocation of the animals. In addition, three mixed anesthesia (volume per unit weight of mouse animal, medetomidine: 0.3 mg / kg, midazolam: 4 mg / kg, butorphanol: 5 mg / kg) was intraperitoneally administered as needed, and the same measures as under isoflurane anesthesia were performed.

[0155] For samples administered to the knee joint, the skin and excess muscle were removed with scissors, the lower limbs were removed from the hip joint, and one by one was collected into a 15 mL tube and frozen for storage. On the day of the Luc activity assay, the frozen specimens were thawed, and the knee joints, skin, knee ligaments, and synovium were collected. For the collection sites of the joint specimens, tissue analysis of the rat joints was performed in advance to identify the collection sites suitable for measuring Luc activity. Specifically, excess muscle was removed from the lower limbs, and the lower end of the femur and the upper end of the tibia were physically crushed with pliers, and the part where the bone marrow components were removed was used as the collection site. The collection site includes at least cartilage, synovium, cruciate ligament, joint capsule, etc.

[0156] For samples administered to the spleen or testis, the entire organ was collected as one specimen and stored frozen at -80° C. In addition, the epididymal caput and epididymal cauda attached to the testis were carefully separated from the testis and stored frozen.

[0157] Regarding the liver, a 1 cm square (about 3 mm thick) was collected centered on the administration site and stored frozen at -80°C.

[0158] (Luc activity assay)

[0159] The Luc activity was determined using a Luc-assay kit (Promega). The attached passive lysis buffer was added to 1 mL of the rat knee joint specimen, 1 mL of the mouse spleen specimen and mouse testis specimen, 0.3 mL each of the mouse epididymal head and epididymal tail specimens, and 0.5 mL of the mouse liver, and chopped with scissors. Then, it was frozen in a freezer, and then thawed at room temperature, and centrifuged at 10000×g, 10 minutes, and 16°C. According to the procedure, 20 μL of the supernatant and 100 μL of the substrate solution were mixed in a Lumitube (Kikkoman), and the relative fluorescence unit (RFU) was quickly measured using Lumiester C-110 (Kikkoman). The RFU measured value was multiplied by the correction value, and the RFU per unit of the total amount of passive lysis buffer added was calculated, which was used as the total RFU per unit tissue specimen.

[0160] [Test Example 1]

[0161] 30 μL of pLuc at a concentration of 1 mg / mL was administered into the knee joint of rats using a 2.5 mL Hamilton syringe with a 30G needle (administration speed 15 μL / s or 0.5 μL / s, each n=2), 30G-pNFI-25 / 40 (n=4) or 30G-pNFI-35 / 40 (n=4). Hereinafter, they are recorded as Syr group, 25 / 40Device group and 35 / 40Device group. The lower limbs were collected 24 hours after administration (day 1) and 10 days after administration (day 10), and the Luc activity in the knee joint was measured. The results of comparison of the mean + SD are shown in Figure 5 In the Syr group, Luc activity (total RFU) was 14150 and 34350 on the first day of "15 μL / s", 11550 and 11800 on the first day of "0.5 μL / s", 400 and 400 on the tenth day of "15 μL / s", and 1450 and 400 on the tenth day of "0. μL / s", which were the same level, so the samples at each collection time point were combined as the Syr group (n=4).

[0162] The Luc activity of the 25 / 40Device group on day 1 was 31.1 times that of the Syr group, and the Luc activity of the 35 / 40Device group on day 1 was 42.3 times that of the Syr group ( Figure 5 ). The Luc activity of the 25 / 40Device group on the 10th day was 32.8 times that of the Syr group, and the Luc activity of the 35 / 40Device group on the 10th day was 184.3 times that of the Syr group ( Figure 5). In all groups, a decrease in Luc activity was observed on day 10 compared to day 1, but the decrease in the 35 / 40Device group was less than that in the Syr group and the 25 / 40Device group. No significant difference was confirmed between the 25 / 40Device group and the 35 / 40Device group on day 1 and day 10, but a tendency was observed that the Luc activity of the 35 / 40Device group was higher.

[0163] [Test Example 2]

[0164] 30 μL of pLuc at a concentration of 1 mg / mL was administered into the knee joint of rats using a 25 mL Hamilton syringe with a 30G needle (dosage rate 0.25 μL / s, n=4 or 83.3 μL / s, n=8) or 30G-pNFI-35 / 40 (n=8). The following are recorded as the 0.25 μL / s administration rate group, the 83.3 μL / s administration rate group, and the 30G-pNFI-35 / 40 group. It should be noted that the ejection rate of the 30G-pNFI-35 / 40 group was 692.8 μL / s as shown in Test Example 4 described later. The lower limbs were collected 24 hours after administration, and the Luc activity in the knee joint was measured. The results of comparing the mean value + SD are shown in Figure 6 .

[0165] In terms of Luc activity, the 83.3 μL / s group and the 30G-pNFI-35 / 40 group were 3.9 times and 105 times higher than those of the 0.25 μL / s group and 105 times higher, respectively. The Luc activity of the 30G-pNFI-35 / 40 group was significantly higher than that of the 0.25 μL / s group and the 83.3 μL / s group.

[0166] [Test Example 3]

[0167] 30 μL of 1 mg / mL pLuc was administered to the knee joint of rats using 30G-pNFI-35 / 40 (n=6), 30G-pNFI-55 / 40 (n=6), or 30G-pNFI-75 / 40 (n=6). Hereinafter, they are respectively recorded as 35 / 40 group, 55 / 40 group, and 75 / 40 group. The lower limbs were collected 24 hours after administration, and the Luc activity in the knee joint was measured. The results of comparison of the mean + SD are shown in Figure 7 .

[0168] No significant differences were observed among the groups in the intra-articular Luc activities of the 35 / 40 group, the 55 / 40 group, and the 75 / 40 group.

[0169] [Test Example 4]

[0170] The injection speed was measured using 30G-pNFI-25 / 40, 30G-pNFI-35 / 40, 30G-pNFI-55 / 40, 30G-pNFI-75 / 40 or pNFI-35 / 40. The tip of the injector was immersed in PBS and 30 μL of 1% malachite green aqueous solution was injected. The time required from the start to the end of injection was measured by high-speed camera analysis, and 30 μL was divided by this time to calculate the injection speed. The results are shown in Figure 8 .

[0171] The injection speed of 30G-pNFI with needle was significantly higher than the maximum speed of the infusion pump of 83.3 μL / s under all conditions, reaching 398.4 μL / s or more. In addition, the injection speed increased depending on the dosage of ZPP.

[0172] [Test Example 5]

[0173] pLuc was administered to the spleen of mice at a concentration of 1 mg / mL using an insulin syringe with a 30G injection needle (dosage volume 30 μL, dosing speed about 30 μL / s, n=4) or 30G-pNFI-35 / 40 (dosage volume 3 μL: n=4, 10 μL: n=4, 30 μL: n=3). Spleens were collected 24 hours after administration and Luc activity was measured. The results of comparison of mean values ​​+ SD are shown in Fig. 9 .

[0174] When 30G-pNFI-35 / 40 was used, Luc activity increased in a volume-dependent manner. In addition, when the same amount of pLuc was administered using 30G-pNFI-35 / 40, Luc activity increased approximately 3 times compared to when an insulin syringe with a 30G injection needle was used.

[0175] [Test Example 6]

[0176] 30 μL of pLuc at a concentration of 0.3 mg / mL, 1 mg / mL, or 3 mg / mL was administered to the spleen of mice using a Hamilton syringe with a 30G needle (administration rate 0.25 μL / s, n=4) or 30G-pNFI-35 / 40 (n=4). Spleens were harvested 24 hours after administration and Luc activity was measured. The results of comparison of mean values ​​+ SD are shown in Fig.10 .

[0177] In the case of 30G-pNFI-35 / 40, the Luc activity of the 1 mg / mL administration group was 3.6 times that of the 0.3 mg / mL administration group, and the Luc activity of the 3 mg / mL administration group was about the same as that of the 1 mg / mL administration group. In addition, the same tendency was confirmed when a Hamilton syringe with a 30G needle (30G-Syr / Pump) was used. When 30G-pNFI-35 / 40 and a Hamilton syringe with a 30G needle were compared at the same pLuc concentration, 30G-pNFI-35 / 40 showed a Luc activity about 50 times higher than that of the Hamilton syringe with a 30G needle at any pLuc concentration.

[0178] [Test Example 7]

[0179] 40 μL of pLuc at a concentration of 1 mg / mL was administered to the spleen of mice using a Hamilton syringe with a 30G needle (administration speed 1 μL / s or 10 μL / s, n=4 for each), 30G-pNFI-25 / 40 (n=4) or 30G-pNFI-35 / 40 (n=4). Spleens were harvested 24 hours after administration and Luc activity was measured. The results of comparison of mean values ​​+ SD are shown in Fig.11 .

[0180] In the case of using 30G-pNFI, Luc activity increased in a manner dependent on the dosing rate (ZPP powder amount). In addition, in the case of a Hamilton syringe with a 30G needle (30G-Syr / pump), Luc activity increased in a manner dependent on the dosing rate.

[0181] [Test Example 8]

[0182] 30 μL of pLuc at a concentration of 1 mg / mL was administered to the testes of mice using a Hamilton syringe with a 30G needle (administration speed 1 μL / s or 10 μL / s, n=4 for each), 30G-pNFI-25 / 40 (n=4) or 30G-pNFI-35 / 40 (n=4). 24 hours after administration, the testes, epididymal caput and epididymal cauda were collected and the Luc activity was measured. The results of the comparison of the mean + SD of the Luc activity of the testes, epididymal caput and epididymal cauda are shown in Fig.12 , Fig.13 , Fig.14 .

[0183] The administration rate was 1 μL / s or 10 μL / s using a Hamilton syringe with a 30G needle, and the RFU values ​​in the testis were 3.0 ± 1.2 × 10 5 , 8.2±1.9×10 5In the case of using 30G-pNFI-25 / 40 and 30G-pNFI-35 / 40, the RFU values ​​in the testis were 6.1 ± 3.7 × 10 7 , 7.3±3.6×10 7 , to the same extent.

[0184] [Test Example 9]

[0185] 10 μL of pLuc at a concentration of 1 mg / mL was administered to the lateral left lobe of the liver of mice using a Hamilton syringe with a 30G needle (administration speed 1 μL / s or 10 μL / s) or 30G-pNFI-25 / 40. The liver was harvested 24 hours after administration and the Luc activity was measured. The results of comparison of the mean + SD (n = 4) are shown in Fig.15 .

[0186] The Luc activity increased in the order of administration using a Hamilton syringe with a 30G needle at an administration rate of 1 μL / s, administration using a Hamilton syringe with a 30G needle at an administration rate of 10 μL / s, and administration using 30G-pNFI-25 / 40. From the above results, it is believed that the Luc activity increases depending on the administration rate.

Claims

1. An injector for injecting a solution containing a functional substance in a living body into an injection target, the injector comprising: A containing part, containing the solution containing the functional substance in the organism; a pressurizing unit, which pressurizes the solution containing the functional substance in the living body when in operation; and an ejection portion, the ejection portion being insertable into the injection object, the ejection portion having an inlet hole and an outlet hole, wherein when the solution containing the in vivo functional substance is pressurized by the pressurizing portion, the inlet hole enables the solution containing the in vivo functional substance to flow in from the containing portion, and the outlet hole enables the inflow of the solution containing the in vivo functional substance to the injection object, The ejection speed of the solution containing the functional substance in the organism is greater than 83.3 μL / s.

2. The injector according to claim 1, wherein: The injection speed of the solution containing the in vivo functional substance is 200 μL / s or more.

3. The injector according to claim 1 or 2, wherein: The inner diameter of the injection portion is less than 0.41 mm.

4. The injector according to any one of claims 1 to 3, wherein: The outer diameter of the emitting portion is less than 0.72 mm.

5. The injector according to any one of claims 1 to 4, wherein: The ejection portion is an injection needle.

6. The injector according to any one of claims 1 to 5, wherein: The in vivo functional substance is one or more selected from the group consisting of nucleic acids, peptides, proteins, and low molecular weight compounds.

7. The injector according to any one of claims 1 to 6, wherein: The injection target is one or more selected from the group consisting of joint cavity, spleen, eyeball, skin, muscle, bone, cartilage, bone marrow, ligament, brain, spinal cord, lung, liver, heart, kidney, pancreas, gallbladder, digestive tract, bladder, reproductive organ, lymph node, lymphatic network and blood vessel, tumor in any of these places, and mucous membrane and connective tissue attached to any of these places. 8 . An injection method, comprising injecting the solution containing the in vivo functional substance into the injection target using the injector according to claim 1 .

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