Liquid pharmaceutical composition

Abstract

The technical problem of the present disclosure at least provides the following contents. Namely, provided is a technique with which it is possible to obtain high physiological activity when a liquid pharmaceutical composition containing an in-vivo functional substance that exhibits physiological activity in an injection subject is injected into the injection subject, the technical problem being solved by a liquid pharmaceutical composition containing an in-vivo functional substance, the osmotic pressure of the liquid pharmaceutical composition is greater than 400 mOsmol / kg and 1000 mOsmol / kg or less, and the liquid pharmaceutical composition is injected into an injection subject by an injector that injects the liquid pharmaceutical composition into the injection subject without passing through an injection needle.

Description

Technical Field

[0001] The present invention relates to a liquid pharmaceutical composition. Background Art

[0002] In recent years, as for medical devices for administering liquid medicine, attention has been paid to needle-free syringes from the viewpoints of countermeasures against needle phobia, pursuit of usability, and reduction of infectious waste. Generally speaking, needle-free syringes are known to be syringes that use compressed gas or spring force as a driving force to squeeze liquid medicine out of the nozzle tip and administer liquid medicine into a living body (Non-patent Document 1). In addition, a needle-free syringe that uses the combustion energy of ignition powder as injection energy has been developed (Patent Document 1).

[0003] The needle-free injector, which uses the combustion energy of the ignition powder as the ejection energy, can deliver the medicine to the nucleus and cytosol because the medicine is squeezed out instantly for administration. From the perspective of drug delivery systems (DDS), the usefulness of the needle-free injector has also attracted much attention, and has been studied in various drugs such as low molecules, peptides, proteins, antibodies, etc. with anticancer effects. In particular, in the field of vaccines and immunity, it has been reported that the needle-free injector can increase antibody titers and induce cellular immunity (non-patent literature 2).

[0004] In the field of drug development, it is well known that the treatment modality of nucleic acid drugs such as DNA and RNA generally requires the delivery of active ingredients to the cytosol of cells, and therefore, electroporation, viral vectors, and the use of lipid nanoparticles (LNP: Lipid NanoParticle) have been studied. In addition, the needle-free injector that uses the combustion energy of the ignition charge as the injection energy has also been studied as a method for delivering DNA and RNA to the cytosol to obtain high drug efficacy (Non-Patent Document 2).

[0005] On the other hand, viral vectors or lipid nanoparticles have the problem of side effects such as allergic reactions that may be induced in the subjects, and technologies that do not use viral vectors or lipid nanoparticles are sought. In addition, electroporation has clinical problems such as producing local toxicity and having to use complex machines. Moreover, in the needle-free injector in which the combustion energy of the ignition powder is used as the ejection energy, in order to further improve the delivery amount of nucleic acid pharmaceuticals to the cytosol, the expression efficiency of the gene, and the expression efficiency of the drug effect, these improvements are being promoted.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: Japanese Patent Application Publication No. 2012-61269

[0009] Non-patent literature

[0010] 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 Summary of the invention

[0011] Problems to be solved by the invention

[0012] The technical problem of the present disclosure is to provide at least the following: That is, to provide a technology capable of obtaining high physiological activity when a liquid pharmaceutical composition containing a biological functional substance that exhibits physiological activity in an injection target is injected into the injection target.

[0013] Solutions for solving problems

[0014] The present inventors have found that the above technical problems can be solved by setting the osmotic pressure of the liquid pharmaceutical composition to a predetermined range when injecting the liquid pharmaceutical composition into a subject using an injector that injects the liquid pharmaceutical composition into the subject without using an injection needle.

[0015] One embodiment of the present disclosure is a liquid pharmaceutical composition, wherein the liquid pharmaceutical composition contains a functional substance in vivo, the osmotic pressure of the liquid pharmaceutical composition is greater than 400mOsmol / kg and less than 1000mOsmol / kg, and the liquid pharmaceutical composition is injected into an injection object using an injector that injects the liquid pharmaceutical composition into the injection object without using an injection needle.

[0016] As a preferred embodiment, the liquid pharmaceutical composition comprises a pharmacologically acceptable agent.

[0017] Furthermore, as a preferred embodiment, the pharmacologically acceptable agent in the liquid pharmaceutical composition is an ionic agent.

[0018] Furthermore, as a preferred embodiment, the ionic reagent in the liquid pharmaceutical composition is a buffer solution using phosphoric acid.

[0019] In addition, as a preferred embodiment, the phosphate buffer in the liquid pharmaceutical composition is phosphate-buffered saline.

[0020] Furthermore, as a preferred embodiment, the ionic agent in the liquid pharmaceutical composition is sodium chloride.

[0021] Furthermore, as a preferred embodiment, the pharmacologically acceptable agent in the liquid pharmaceutical composition is a non-ionic agent.

[0022] Furthermore, as a preferred embodiment, the nonionic agent in the liquid pharmaceutical composition is glucose.

[0023] Furthermore, as a preferred embodiment, the liquid pharmaceutical composition is injected into the injection target by an injector that injects the liquid pharmaceutical composition into the injection target without using an injection needle, and is injected into the injection target by jet injection.

[0024] Furthermore, as a preferred embodiment, the in vivo functional substance in the liquid pharmaceutical composition is a nucleic acid.

[0025] Another embodiment of the present disclosure is an injector that injects a liquid medical composition into an injection object without using an injection needle, the injector comprising: a container that contains the liquid medical composition; a nozzle that is connected to the container and has an ejection port for ejecting the liquid medical composition toward the injection object; and a pressurizing portion that pressurizes the liquid medical composition contained in the container during operation, thereby ejecting the liquid medical composition from the ejection port toward the injection object, wherein the liquid medical composition contains a functional substance in vivo and has an osmotic pressure greater than 400 mOsmol / kg and less than 1000 mOsmol / kg.

[0026] In addition, another embodiment of the present disclosure is a method for injecting a liquid pharmaceutical composition into an injection object, comprising: injecting the liquid pharmaceutical composition into the injection object using an injector that injects the liquid pharmaceutical composition into the injection object without using an injection needle, wherein the liquid pharmaceutical composition contains a functional substance in vivo and has an osmotic pressure greater than 400mOsmol / kg and less than 1000mOsmol / kg.

[0027] Effects of the Invention

[0028] The present disclosure can achieve at least the following effects: That is, it can achieve the following effect: it can provide a technology that can obtain high physiological activity when a liquid pharmaceutical composition containing a biological functional substance that exerts physiological activity in an injection target is injected into the injection target. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a diagram showing a schematic structure of a syringe according to one embodiment of the present disclosure.

[0030] Figure 2 It is an image (photograph instead of a drawing) showing the results of Test Example 1 and Comparative Test Example 1 according to one embodiment of the present disclosure.

[0031] Figure 3It is an image (photograph in place of the drawing) showing the results of Test Example 2 and Comparative Test Example 2 according to one embodiment of the present disclosure.

[0032] Figure 4 It is an image (photograph instead of a drawing) showing the results of Test Example 3 and Comparative Test Example 3 according to one embodiment of the present disclosure.

[0033] Figure 5 It is an image (photograph in place of the drawing) showing the results of Test Example 4 of one embodiment of the present disclosure.

[0034] Figure 6 It is an image (photograph instead of a drawing) showing the results of Test Example 5 and Comparative Test Example 5 according to one embodiment of the present disclosure. DETAILED DESCRIPTION

[0035] 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 within the scope of the main purpose 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.

[0036] One embodiment of the present disclosure is a liquid pharmaceutical composition, wherein the liquid pharmaceutical composition contains a functional substance in vivo, the osmotic pressure of the liquid pharmaceutical composition is greater than 400mOsmol / kg and less than 1000mOsmol / kg, and the liquid pharmaceutical composition is injected into an injection object using an injector that injects the liquid pharmaceutical composition into the injection object without using an injection needle.

[0037] (Functional substances in living organisms)

[0038] The in vivo functional substance contained in the liquid pharmaceutical composition of this embodiment is a substance that exerts physiological activity in the injected object. The in vivo functional substance contained in the liquid pharmaceutical composition can be one or more. The in vivo functional substance can be a natural substance or an artificially synthesized in vivo functional substance.

[0039] Examples of the in vivo functional substance include nucleic acids, peptides, proteins, and the like.

[0040] When the in vivo functional substance is a nucleic acid, the nucleic acid may be DNA or RNA. In addition, the nucleic acid may be a nucleic acid containing a protein-coding portion or a nucleic acid not containing a protein-coding portion (non-coding nucleic acid).

[0041] Examples of the case where the nucleic acid includes a portion encoding a protein include the following.

[0042] For example, when the in vivo functional substance is a DNA containing a gene, it exhibits physiological activity as a template for mRNA synthesis (transcription) for producing a protein, and its physiological activity can be quantitatively evaluated by the amount of mRNA and protein produced.

[0043] Furthermore, when the in vivo functional substance is mRNA, it exhibits physiological activity as a template for protein synthesis (translation), and its physiological activity can be quantitatively evaluated by the amount of protein produced.

[0044] In addition, the physiological activity can also be evaluated based on the changes in the injection object caused by the increase or decrease of the mRNA and protein produced. For example, the injection object is an injection object (such as an individual (organism), etc.) that can induce cellular immunity, and the nucleic acid is a nucleic acid encoding a protein that induces cellular immunity. By inducing the increase of the protein of the cellular immunity, in the case of such a change that cellular immunity is induced in the injection object, the nucleic acid exerts a physiological activity such as inducing cellular immunity, and its physiological activity can be quantitatively evaluated by the induction amount of the cellular immunity of the injection object. In addition, it can also be evaluated by the induction amount of the cellular immunity of the injection object after the restimulation (resensitization) caused by the protein that induces the cellular immunity.

[0045] Examples of the case where the nucleic acid does not contain a portion encoding a protein include the following.

[0046] For example, when the injection object is an individual (organism) to which a vaccine can be administered, if the nucleic acid is a DNA containing a CpG motif (CpG DNA) or polyinosinic acid-polycytidylic acid (PolyI:C), it exerts a physiological activity as an adjuvant for enhancing the effect of the vaccine, and its physiological activity can be quantitatively evaluated by anti-tumor effect (the effect can be evaluated, for example, by a reduction in tumor size), maturity of immune cells, etc.

[0047] Furthermore, when the nucleic acid is a small interfering RNA (siRNA), it exhibits physiological activity of degrading mRNA and sequence-specifically inhibiting the expression of a target gene, and its physiological activity can be quantitatively evaluated based on the amount of mRNA or protein produced by the target gene.

[0048] Furthermore, when the nucleic acid is antisense DNA or antisense RNA, it hybridizes with RNA of the target gene and exerts physiological activities such as inhibiting splicing or inhibiting translation, and the physiological activity can be quantitatively evaluated based on the amount of mRNA or protein produced by the target gene.

[0049] The functional substance in the organism in this embodiment exerts physiological activity in the injection object when injected into the injection object and exists stably. In addition, if there is no adverse effect such as destroying the injection object, it can be in a free form or in a form fixed on a carrier such as nanoparticles, or it can be modified, including a solvent, and its scheme is not particularly limited.

[0050] In the examples described below, as the in vivo functional substance, there are listed: an example in which free plasmid DNA containing a GFP (Green Fluorescence Protein) gene is used and the GFP gene is used as a reporter gene; and an example in which mRNA encoding GFP as a reporter protein is used.

[0051] When the DNA comprises a gene, it can be designed in the form of an expression cassette or an expression vector comprising the gene. Moreover, for example, the gene can also be configured under the control of a promoter suitable for the injection object and the injection site of the DNA to be injected. That is, known genetic engineering methods can be used in any scheme. For example, in the embodiments described later, an embodiment using a vector CMV-DASHER-GFP (manufactured by ATUM Co., Ltd.) comprising a GFP gene as an expression vector is listed. The plasmid vector is well known and can be obtained by those skilled in the art. The subcloning of expression vectors and recombinant vectors can be carried out according to known methods.

[0052] When the functional substance in the body is a peptide or a protein, examples of the peptide or protein include: antigens (i.e., substances that cause the production of antibodies against the peptide or protein), antibodies, peptide vaccines, protein vaccines, peptide hormones, protein hormones, growth factors, cytokines, coagulation factors, serum albumin, digestive enzymes, anti-inflammatory peptides, anti-inflammatory proteins, antibiotics, etc.

[0053] In this embodiment, “high physiological activity can be obtained when a liquid pharmaceutical composition containing a biofunctional substance that exerts a physiological activity in a subject is injected into the subject” means that, when evaluating the physiological activity exerted by the biofunctional substance using a quantitative index,

[0054] Compared to the physiological activity (the physiological activity may be, for example, an amount indicating the physiological activity) exerted by the in vivo functional substance when a liquid pharmaceutical composition containing the in vivo functional substance and having an osmotic pressure not within the prescribed range described below is injected into an injection subject using an injector for injecting the liquid pharmaceutical composition into the injection subject without using an injection needle,

[0055] When a liquid pharmaceutical composition containing a biofunctional substance and having an osmotic pressure within a predetermined range described below is injected in the same manner as described above, the biofunctional substance exhibits a large physiological activity (the physiological activity may be, for example, an amount indicating the physiological activity).

[0056] That is, for example,

[0057] When a liquid pharmaceutical composition containing a biofunctional substance and having an osmotic pressure of 400 mOsmol / kg or less or greater than 1000 mOsmol / kg is injected into a subject using an injector that injects the liquid pharmaceutical composition into the subject without using an injection needle (this is referred to as scenario B), the physiological activity exerted by the biofunctional substance is referred to as “activity B”,

[0058] When the same conditions as in Scheme B are used except that the in vivo functional substance is contained and the osmotic pressure is greater than 400 mOsmol / kg and less than 1000 mOsmol / kg, and the liquid pharmaceutical composition is injected into the injection subject in the same manner as above (this is referred to as Scheme A), and the physiological activity exerted by the in vivo functional substance is referred to as “Activity A”,

[0059] Activity B < Activity A.

[0060] In addition, in the present embodiment, “high physiological activity can be obtained when a liquid pharmaceutical composition containing a biofunctional substance that exerts a physiological activity in a subject to be injected is injected into the subject to be injected” may mean that, when the physiological activity exerted by the biofunctional substance is evaluated by a quantitative index,

[0061] Compared with the physiological activity (the physiological activity may be, for example, the amount indicating the physiological activity) exerted by the biofunctional substance when a liquid pharmaceutical composition containing the biofunctional substance and having an osmotic pressure within the predetermined range described below is injected into an injection subject using a syringe with a needle,

[0062] When a liquid pharmaceutical composition containing a biological functional substance and having an osmotic pressure within a specified range described below is injected into an injection target in the same manner as described above, except that the liquid pharmaceutical composition is injected into the injection target without passing through an injection needle, the physiological activity exerted by the biological functional substance (the physiological activity may be, for example, an amount representing the physiological activity) is large.

[0063] That is, for example,

[0064] When a liquid pharmaceutical composition containing a biofunctional substance and having an osmotic pressure of more than 400 mOsmol / kg and less than 1000 mOsmol / kg is injected into an injection subject using a needle syringe (this is referred to as scenario C), the physiological activity exerted by the biofunctional substance is referred to as “activity C”,

[0065] When the same conditions as those of Scheme C are set except that a liquid pharmaceutical composition containing a biofunctional substance and having an osmotic pressure of more than 400 mOsmol / kg and less than 1000 mOsmol / kg is injected into a subject using an injector that injects the liquid pharmaceutical composition into the subject without an injection needle (this is referred to as Scheme A), and the physiological activity exerted by the biofunctional substance is referred to as “Activity A”,

[0066] Activity C < Activity A.

[0067] When the physiological activity of the in vivo functional substance is evaluated by a quantitative index (eg, expression level, antibody titer, disease effectiveness score, etc.), the quantitative index may be appropriately set according to the physiological activity of the in vivo functional substance.

[0068] As for the quantitative index, for example, when the in vivo functional substance is DNA and contains a gene, examples thereof include the amount of mRNA produced using it as a template, the amount of protein produced using the mRNA as a template (usually evaluated as the expression level of the gene), etc.

[0069] Furthermore, for example, when the in vivo functional substance is mRNA, examples thereof include the amount of protein produced using the mRNA as a template.

[0070] That is, for example, when the in vivo functional substance is DNA and contains a gene, and when the quantitative index is the expression level of the gene,

[0071] When a liquid pharmaceutical composition having an osmotic pressure of 400 mOsmol / kg or less or greater than 1000 mOsmol / kg is injected into a subject using an injector that injects the liquid pharmaceutical composition into the subject without using an injection needle (this is referred to as scenario B1), the expression level of the gene is referred to as “expression level B1”,

[0072] When the same conditions as in Scheme B1 are used except that the osmotic pressure is set to be greater than 400 mOsmol / kg and less than 1000 mOsmol / kg, and the liquid pharmaceutical composition is injected into the injection subject in the same manner as above (this is referred to as Scheme A1), and the expression level of the gene is referred to as “expression level A1”,

[0073] Expression level B1<expression level A1.

[0074] Alternatively, when a liquid pharmaceutical composition having an osmotic pressure of more than 400 mOsmol / kg and less than 1000 mOsmol / kg is injected into an injection subject using a needle syringe (this is referred to as protocol C1), the expression level of the gene may be referred to as “expression level C1”.

[0075] When the same conditions as those of Scheme C1 (Scheme A1) are used except that a liquid pharmaceutical composition having an osmotic pressure of greater than 400 mOsmol / kg and less than 1000 mOsmol / kg is injected into the injection subject using an injector that injects the liquid pharmaceutical composition into the injection subject without an injection needle, and the expression level of the gene is referred to as “expression level A1”,

[0076] Expression level C1<expression level A1.

[0077] When the in vivo functional substance is a peptide or a protein, the quantitative index may include, for example, the following.

[0078] For example, when the injection object is an injection object that can produce antibodies (such as an individual (organism)), and the peptide or protein is a substance (antigen, etc.) that causes antibodies to the peptide or protein to be produced in the injection object, for example, the antibody titer of the antibody can be listed.

[0079] In addition, for example, when the injection object is an injection object that can induce cellular immunity (for example, an individual (organism), etc.), and the peptide or protein is a substance that induces cellular immunity in the injection object, examples include: the amount of cytokine production, the number of cytokine-producing cells measured by ELISpot, etc.

[0080] In addition, for example, when the injection object is an injection object that can activate immune cells (for example, an individual (organism), etc.), and the peptide or protein is a substance that activates immune cells in the injection object, examples include: the production amount of CD80, the production amount of CD86, the production amount of CD40, the production amount of MHC molecules, the total number of immune cells, etc.

[0081] Furthermore, for example, when the injection target is a disease model (e.g., an individual (organism) etc.) and the peptide or protein is a substance that improves the disease in the injection target, examples include: a value of the effectiveness score for the disease etc.

[0082] As a method for evaluating the quantitative index, for example, after injecting the liquid pharmaceutical composition into an injection subject, the method can be appropriately selected according to the physiological activity exerted by the in vivo functional substance using all or part of the injection subject.

[0083] For example, as in the embodiments described later, when the injection object is an individual (organism), the functional substance in the organism is a DNA containing a gene, and the expression level of the gene in the vicinity of the injection site (i.e., a part of the injection object) is evaluated, after the liquid medical composition is injected into the injection object, the vicinity of the injection site is obtained and appropriately selected according to the physiological activity exerted by the functional substance in the organism.

[0084] In addition, as described in the examples below, samples can be prepared by known methods, and the physiological activity exerted by the functional substance in the organism can be evaluated by the amount of protein produced. In the examples described below, since the protein produced is GFP protein, the physiological activity is evaluated by fluorescence brightness as an indicator.

[0085] In addition, for example, when the injection target is an individual (organism), the functional substance in the organism is a nucleic acid encoding a protein that induces cellular immunity, and the induction amount of cellular immunity is to be evaluated, after the liquid pharmaceutical composition is injected into the injection target, the induction amount of cellular immunity can be evaluated using the individual (organism) itself (i.e., the entire injection target) and using a known method for evaluating the induction amount of cellular immunity.

[0086] (Osmotic pressure)

[0087] If the osmotic pressure of the liquid pharmaceutical composition of this embodiment is within a predetermined range, when the liquid pharmaceutical composition is injected into a subject using an injector that injects the liquid pharmaceutical composition into the subject without using an injection needle, higher physiological activity can be obtained compared to the case of injecting a conventional liquid pharmaceutical composition.

[0088] The lower limit of the prescribed range is, for example, greater than 400 mOsmol / kg, 436 mOsmol / kg or more, 460 mOsmol / kg or more, 584 mOsmol / kg or more, 605 mOsmol / kg or more, 624 mOsmol / kg or more, 872 mOsmol / kg or more, or 879 mOsmol / kg or more.

[0089] It is speculated that: when the injection target is a cell or an injection target containing a cell, the osmotic pressure is relatively large, and the difference between the osmotic pressure of the liquid pharmaceutical composition and the osmotic pressure in the cell easily generates physical stress on the cell membrane, and the substance permeability of the cell membrane is easily improved, and as a result, the amount of functional substances injected into the cell in the body is easily increased, and high physiological activity is easily obtained. On the contrary, it is speculated that: when the osmotic pressure is relatively small, the difference between the osmotic pressure of the liquid pharmaceutical composition and the osmotic pressure in the cell is difficult to increase (or no difference is generated), and it is difficult to generate physical stress on the cell membrane, and it is difficult to affect the substance permeability of the cell membrane. As a result, the amount of functional substances injected into the cell in the body is also difficult to increase, and it is difficult to obtain high physiological activity.

[0090] In addition, it is speculated that when the injection target is, for example, an individual (organism), the osmotic pressure is high, and the local retention of the liquid pharmaceutical composition injected into the injection target is likely to be improved, and as a result, high physiological activity is easily obtained in the injection target. On the contrary, it is speculated that when the osmotic pressure is low, the local retention of the liquid pharmaceutical composition injected into the injection target is likely to be reduced, and the liquid pharmaceutical composition is likely to be decomposed and dispersed, and as a result, it is difficult to obtain high physiological activity in the injection target.

[0091] In addition, the upper limit of the specified range is, for example, 1000 mOsmol / kg or less, 899 mOsmol / kg or less, 879 mOsmol / kg or less, 872 mOsmol / kg or less, 624 mOsmol / kg or less, 605 mOsmol / kg or less, 584 mOsmol / kg or less, 460 mOsmol / kg or less, or 436 mOsmol / kg or less.

[0092] The lower the osmotic pressure, the less likely it is that the load on the object of injection caused by the injection of the liquid pharmaceutical composition will be suppressed as much as possible. For example, when the object of injection is a cell or contains cells, cell damage or death can be suppressed as much as possible. In addition, for example, when the object of injection is a mammalian individual (organism), the occurrence of internal bleeding at the injection site caused by the injection of the liquid pharmaceutical composition can be suppressed as much as possible.

[0093] In view of the reason for adopting the upper limit and the reason for adopting the lower limit, the upper limit and the lower limit of the prescribed range may be a non-contradictory combination selected from the above-mentioned upper limit examples and lower limit examples.

[0094] For example, it may be greater than 400mOsmol / kg and less than 1000mOsmol / kg, greater than 400mOsmol / kg and less than 899mOsmol / kg, greater than 400mOsmol / kg and less than 879mOsmol / kg, greater than 400mOsmol / kg and less than 872mOsmol / kg, greater than 400mOsmol / kg and less than 624mOsmol / kg, greater than 400mOsmol / kg and less than 605mOsmol / kg, greater than 400mOsmol / kg and less than 584mOsmol / kg, greater than 400mOsmol / kg and less than 460mOsmol / kg, or greater than 400mOsmol / kg and less than 436mOsmol / kg.

[0095] In addition, for example, it can be 436mOsmol / kg to 1000mOsmol / kg, 436mOsmol / kg to 899mOsmol / kg, 436mOsmol / kg to 879mOsmol / kg, 436mOsmol / kg to 872mOsmol / kg, 436mOsmol / kg to 624mOsmol / kg, 436mOsmol / kg to 605mOsmol / kg, 436mOsmol / kg to 584mOsmol / kg, or 436mOsmol / kg to 460mOsmol / kg.

[0096] In addition, for example, it can be 460mOsmol / kg to 1000mOsmol / kg, 460mOsmol / kg to 899mOsmol / kg, 460mOsmol / kg to 879mOsmol / kg, 460mOsmol / kg to 872mOsmol / kg, 460mOsmol / kg to 624mOsmol / kg, 460mOsmol / kg to 605mOsmol / kg, or 460mOsmol / kg to 584mOsmol / kg.

[0097] In addition, for example, it can be 584mOsmol / kg to 1000mOsmol / kg, 584mOsmol / kg to 899mOsmol / kg, 584mOsmol / kg to 879mOsmol / kg, 584mOsmol / kg to 872mOsmol / kg, 584mOsmol / kg to 624mOsmol / kg, or 584mOsmol / kg to 605mOsmol / kg.

[0098] For example, it may be 605 mOsmol / kg to 1000 mOsmol / kg, 605 mOsmol / kg to 899 mOsmol / kg, 605 mOsmol / kg to 879 mOsmol / kg, 605 mOsmol / kg to 872 mOsmol / kg, or 605 mOsmol / kg to 624 mOsmol / kg.

[0099] For example, it may be 624 mOsmol / kg to 1000 mOsmol / kg, 624 mOsmol / kg to 899 mOsmol / kg, 624 mOsmol / kg to 879 mOsmol / kg, or 624 mOsmol / kg to 872 mOsmol / kg.

[0100] Furthermore, for example, it may be 872 mOsmol / kg to 1000 mOsmol / kg, 872 mOsmol / kg to 899 mOsmol / kg, or 872 mOsmol / kg to 879 mOsmol / kg.

[0101] Furthermore, for example, it may be 879 mOsmol / kg to 1000 mOsmol / kg, or 879 mOsmol / kg to 899 mOsmol / kg.

[0102] The osmotic pressure of the liquid pharmaceutical composition can be measured using, for example, Osmometer 3000 Basic manufactured by Gonotec.

[0103] (Injection object)

[0104] The injection object in this embodiment can be, for example, one or more selected from the group consisting of cells, cell sheets, cell clusters, tissues, organs (skin, organs, etc.), organ systems, individuals (organisms), etc., without limitation. It can be any type represented by an in vitro system, an in vivo system, or an ex vivo system. The cell cluster can be a cell cluster obtained by three-dimensional culture, and the organ (skin, organs, etc.) can be an organoid.

[0105] In addition, when the injection object is injected, it is also possible to inject into the lower level contained in the object. That is, for example, when an individual (organism) is used as the injection object, it is possible to inject into the tissue contained in the individual (organism), it is also possible to inject into the cells contained in the individual (organism), or it is also possible to inject into both. In addition, when a tissue is used as the injection object, it is possible to inject into the cells contained in the tissue, it is also possible to inject into the intercellular matrix contained in the tissue, or it is also possible to inject into both.

[0106] In addition, when the injection object in the present embodiment is one or more selected from the group consisting of cells, cell sheets, cell clusters, tissues, organs (skin, organs, etc.), and organ systems, it can be one or more selected from the group consisting of cells, cell sheets, cell clusters, tissues, organs (skin, organs, etc.), and organ systems in a state existing in an individual (organism), or it can be one or more selected from the group consisting of cells, cell sheets, cell clusters, tissues, organs (skin, organs, etc.), and organ systems in a state not existing in an individual (organism) (for example, a state of being removed and separated from an individual (organism), or a state of being produced outside an individual (organism)).

[0107] In addition, the injection object in this embodiment can also be selected from one or more of the group consisting of cells, cell sheets, cell clusters, tissues, organs (skin, organs, etc.), and organ systems derived from stem cells such as iPS cells (artificial pluripotent stem cells), which can be present in an individual (organism) or not in an individual (organism) (for example, a state of being removed and separated from an individual (organism), or a state of being made outside an individual (organism)). The cell cluster can be a cell cluster obtained by three-dimensional culture, and the organ (skin, organ, etc.) can be an organoid.

[0108] The individual (organism) is preferably a mammalian individual (organism). The mammal is not particularly limited, and humans can be listed, and mammals other than humans can be listed. Mammals other than humans include mice, rats, guinea pigs, hamsters, cows, goats, sheep, pigs, monkeys, dogs, cats, etc.

[0109] In addition, no matter which of the above-mentioned injection targets is used in this embodiment, when injected into cells, it can be injected into the cytoplasm of the cell, into the nucleus of the cell, or into both the cytoplasm and the nucleus of the cell.

[0110] In addition, the injection object in this embodiment is not particularly limited, and is preferably one or more selected from the group consisting of the intradermal, subcutaneous, and muscle layers below the skin of a mammalian individual (organism). In this case, the liquid pharmaceutical composition can be injected from an injector toward the skin surface and injected into the skin, and injected into the skin, and one or more selected from the group consisting of the intradermal, subcutaneous, and muscle layers below the skin.

[0111] (Liquid pharmaceutical composition)

[0112] The liquid pharmaceutical composition of the present embodiment may be mixed with a non-toxic carrier for pharmaceutical composition in addition to the biological functional substance. Examples thereof include substances described in the description of the pharmacologically acceptable reagents described later. In addition, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, isotonic agents, etc. may be appropriately added as needed.

[0113] The content of the in vivo functional substance relative to the total amount of the liquid pharmaceutical composition of this embodiment can be appropriately set based on the type of the in vivo functional substance, the injection object, the physiological activity exerted by the in vivo functional substance in the injection object injected with the in vivo functional substance, etc. In addition, when the liquid pharmaceutical composition contains a pharmacologically acceptable agent described later, the content of the pharmacologically acceptable agent can also be appropriately set in consideration.

[0114] In addition, the injection amount of the liquid pharmaceutical composition into the injection object can be appropriately set based on the content or type of the biological functional substance, the injection object, the physiological activity exerted by the biological functional substance in the injection object injected with the biological functional substance, etc. In addition, in the case where the liquid pharmaceutical composition contains a pharmacologically acceptable agent described later, the injection amount of the pharmacologically acceptable agent can also be appropriately set in consideration.

[0115] The pH of the liquid pharmaceutical composition of the present embodiment is not particularly limited as long as the functional substance in the body exerts physiological activity and stably exists in the injected object when the liquid pharmaceutical composition is injected into the injected object using an injector that injects the liquid pharmaceutical composition into the injected object without passing through an injection needle, and there is no adverse effect such as damaging the injected object.

[0116] The liquid pharmaceutical composition of this embodiment may be in the form of a vaccine.

[0117] (Pharmacologically acceptable agent)

[0118] The liquid pharmaceutical composition of this embodiment preferably contains a pharmacologically acceptable agent. As for the pharmacologically acceptable agent, the scheme is not particularly limited as long as the functional substance in the body exerts physiological activity and stably exists in the injected object when the liquid pharmaceutical composition is injected into the injected object using an injector that injects the liquid pharmaceutical composition into the injected object without an injection needle, and there is no adverse effect such as damaging the injected object.

[0119] The content of the pharmacologically acceptable agent relative to the total amount of the liquid pharmaceutical composition of this embodiment can be appropriately set based on the content or type of the biological functional substance, the injection target, the physiological activity exerted by the biological functional substance in the injection target, etc.

[0120] The pharmacologically acceptable agent may be an agent for adjusting the osmotic pressure of the liquid pharmaceutical composition. In addition, when the pharmacologically acceptable agent is a solution, it may be a solvent for the liquid pharmaceutical composition or a solvent for dissolving the functional substance in the organism. In addition, when the pharmacologically acceptable agent is a solid, it may be a solvent for the liquid pharmaceutical composition in the form of a pharmacologically acceptable liquid (e.g., water (e.g., water for injection, etc.)) or a solvent for dissolving the functional substance in the organism.

[0121] It should be noted that, in the present disclosure, “dissolve” may be “suspend” or “emulsify”, “solution” may be “suspension” or “emulsion”, and “solvent” may be “suspending agent” or “emulsifier”.

[0122] As the pharmacologically acceptable agent, ionic agents, nonionic agents, and the like are preferred from the viewpoint of compatibility with the injection subject.

[0123] Among them, ionic reagents are preferred from the viewpoint of versatility in commercial use.

[0124] Furthermore, from the viewpoint of storage stability such as freeze-drying, nonionic reagents are preferred.

[0125] Among the ionic reagents, known ionic buffers can be used as ionic buffers. 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.), citric acid buffer, tris-HCl buffer (Tris hydrochloric acid 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.

[0126] In addition, from the viewpoint of the actual use effect of the pharmaceutical, known ionic additives for pharmaceuticals can be used. For example, sodium chloride, sodium ascorbate, magnesium sulfate, sodium oleate, sodium acetyltryptophanate, amino acids, calcium citrate, monosodium succinate, sodium tartrate, aluminum hydroxide, phospholipids, sodium thioglycolate, sulfuric acid, sodium sulfate, hydrochloric acid, sodium hydrochloride, sodium hydroxide, pH adjusters (such as hydrochloric acid, sulfuric acid, sodium hydroxide, etc.), phosphoric acid, sodium phosphate, sodium dihydrogen phosphate, etc. can be used.

[0127] Furthermore, from the viewpoint of versatility, a buffer solution using sodium chloride or phosphoric acid is preferred.

[0128] As the nonionic agent, known nonionic additives for pharmaceuticals can be used, for example, glucose, mannose, mannitol, sorbitol, trehalose, sucrose, glycerol, polyethylene glycol, cholesterol, tocopherol, cyclodextrin, hydroxypropylmethylcellulose, etc. can be used.

[0129] Furthermore, from the viewpoint of the actual use effect of the pharmaceutical product, glucose, mannose, mannitol, sorbitol, trehalose, sucrose, glycerol, and polyethylene glycol are preferred.

[0130] Furthermore, from the viewpoint of versatility, glucose is preferred.

[0131] (Injector)

[0132] As the injector in this embodiment, an injector capable of injecting the liquid pharmaceutical composition into the injection object without using an injection needle can be used. That is, the liquid pharmaceutical composition can be injected into the injection object using an injector capable of injecting the liquid pharmaceutical composition into the injection object without using an injection needle.

[0133] In the injector of this embodiment, the "top side" refers to the side where the ejection port for ejecting the liquid pharmaceutical composition from the injector is arranged, and the "base side" refers to the side opposite to the top side in the injector, and these expressions do not limit the specific parts or positions.

[0134] In the injector in the present embodiment, in order to eject the liquid medical composition to the injection object, the driving unit imparts the injection energy. The "injection" achieved by the injector in the present embodiment is achieved by pressurizing the liquid medical composition contained in the containing part by the pressurizing part using the injection energy imparted by the driving unit, thereby causing the liquid medical composition to flow through the flow path of the containing part. As the injection energy, it can be the injection energy used in the previous injector, for example, the combustion energy of gunpowder, the generation energy of gas generating agent, the electrical energy of piezoelectric element, the mechanical energy of spring, etc., and in addition, the energy formed by appropriately combining these forms of energy can be used.

[0135] When the combustion energy of the gunpowder is used as the injection energy, the gunpowder includes, for example, a gunpowder containing zirconium and potassium perchlorate (ZPP), a gunpowder containing titanium hydride and potassium perchlorate (THPP), a gunpowder containing titanium and potassium perchlorate (TiPP), a gunpowder containing aluminum and potassium perchlorate (APP), a gunpowder containing aluminum and bismuth oxide (ABO), a gunpowder containing aluminum and molybdenum oxide (AMO), a gunpowder containing aluminum and copper oxide (ACO), a gunpowder containing aluminum and iron oxide (AFO), or a gunpowder containing a combination of multiple thereof. As a feature of these gunpowders, even if the combustion product thereof is gaseous at high temperature, it does not contain gaseous components at room temperature, so the combustion product condenses immediately after ignition. Thus, in the pressurization process for injecting the liquid pharmaceutical composition, the temperature of the combustion product during pressurization can be shifted to near room temperature in a short time after the pressure applied to the liquid pharmaceutical composition reaches the first peak injection force by the combustion of the ignition powder.

[0136] In addition, when the generation energy of the gas generating agent is used as the injection energy, a single-base smokeless gunpowder (for example, a single-base smokeless gunpowder containing 98 mass % of nitrocellulose, 0.8 mass % of diphenylamine, and 1.2 mass % of potassium sulfate) or various gas generating agents used in gas generators for air bags and gas generators for seat belt pretensioners can also be used as the gas generating agent.

[0137] In the injector of this embodiment, the pressurizing portion pressurizes the liquid medical composition contained in the containing portion during operation, thereby causing the liquid medical composition to be ejected from the ejection port toward the injection target.

[0138] The pressurization by the pressurizing unit is not particularly limited as long as it does not damage the system, such as damaging the housing unit, and the pressurization conditions of a normal injector can be adopted.

[0139] The pressure refers to the pressure inside the receiving part. There is no particular limitation on the measuring method, and for example, the measurement can be performed as described below. That is, as in the measuring method described in Japanese Patent Gazette No. 2005-21640, the measurement is performed by the following method: the injection force is dispersedly applied to the diaphragm of the force sensor arranged downstream of the nozzle, and the output from the force sensor is collected by a data acquisition device via a detection amplifier and stored as the injection force (N) according to time. The injection pressure is calculated by dividing the injection pressure measured in this way by the area of ​​the injection port 31a of the injector. The measured value of the internal pressure of the receiving part is the same as the injection pressure, and the injection pressure can be used as the pressure inside the receiving part.

[0140] The injection energy obtained by the driving unit is transmitted to the plunger via the piston, causing the plunger to slide in the container, thereby squeezing the liquid medical composition contained in the container along the flow path formed in the nozzle unit and finally ejected from the ejection port toward the injection target.

[0141] The container may contain the liquid pharmaceutical composition from the beginning, or may not contain the liquid pharmaceutical composition. When not contained, the liquid pharmaceutical composition is contained by sucking it into the container through a nozzle having an ejection port. In this way, by adopting a structure that requires a containing operation of the container, any desired liquid pharmaceutical composition can be injected into the injection object. Therefore, in the injector in this embodiment, the syringe part and the injector body are configured to be detachable.

[0142] In this embodiment, the liquid pharmaceutical composition may be injected into the injection target by using an injector that injects the liquid pharmaceutical composition into the injection target without using an injection needle, and may be injected into the injection target by jet injection.

[0143] Here, in the present disclosure, "jet injection" is an injection that is not performed via an injection needle, and the injection is characterized in that a liquid pharmaceutical composition containing a functional substance in a living body and having an osmotic pressure greater than 400mOsmol / kg and less than 1000mOsmol / kg is ejected from an ejection port toward an injection object, thereby forming a through hole that penetrates the inner and outer boundaries of the injection object, and generating a high-pressure ultrafine liquid flow that can pass through the through hole and inject the liquid pharmaceutical composition into the injection object. For example, in the case where the injection object is a mammalian individual (organism), the injection refers to an injection characterized by generating a high-pressure ultrafine liquid flow that can penetrate the skin of the mammalian individual (organism).

[0144] The injector in the present embodiment can be obtained as follows: in a previous injector that can be used to inject an injection (liquid pharmaceutical composition) into an injection object without an injection needle, the liquid pharmaceutical composition is contained in the containing portion. As such a previous injector, for example, an injector described in International Publication No. 2019 / 156238, an injector described in International Publication No. 2019 / 156239, an injector described in International Publication No. 2019 / 156237, an injector described in the specification of Japanese Patent No. 5989039, etc. can be cited. They are injectors that can perform the jet injection, but in addition, there are many commercially available injectors (needle-free syringes) that can perform the jet injection. For example, Straits (Pharmajet), Tropis (Pharmajet), Vitajet (Bioject Medical Technologies Inc.), Biojector2000 (Bioject Medical Technologies Inc.), BiojectZetajet (Bioject Medical Technologies Inc.), Glide (Glide Pharma), MediJectorVision (Antares), Sumaval DosePro (Zogenix), SQ Pen (Bespak), Injex (Equidyne), hyaluronic acid injection cartridge (BEAUTTO, Amazon Standard Identification Number (ASIN): B08NCHTRHZ), etc.

[0145] Hereinafter, with reference to the accompanying drawings, a syringe 1 (needle-free syringe) will be described as an example of an injector of the present embodiment. It should be noted that the configuration of the following embodiment is an example and is not limited to the configuration of the present embodiment. It should be noted that "top end side" and "base end side" are used as terms to indicate the relative positional relationship in the length direction of the syringe 1. The "top end side" indicates a position close to the top end of the syringe 1 described later, i.e., close to the ejection port 31a, and the "base end side" indicates a direction on the opposite side of the "top end side" in the length direction of the syringe 1, i.e., the direction on the drive unit 7 side. In addition, this example is an example in which the combustion energy of the gunpowder ignited by the ignition device is used as the ejection energy for pressurization, but the present embodiment is not limited to this.

[0146] (Structure of the syringe 1)

[0147] Figure 11 is a diagram showing a schematic structure of the syringe 1, and is also a cross-sectional view of the syringe 1 along its longitudinal direction. The syringe 1 is constructed as follows: a syringe assembly 10 is mounted on a housing (syringe housing) 2, and the syringe assembly 10 is formed by integrally assembling a subassembly consisting of a syringe barrel 3 and a plunger 4 and a subassembly consisting of a syringe body 6, a piston 5, and a drive unit 7.

[0148] As described above, the syringe assembly 10 is configured to be detachable from the housing 2. The liquid medical composition is filled in the housing 32 formed between the syringe barrel 3 and the plunger 4 included in the syringe assembly 10, and the syringe assembly 10 is a unit that is used up and thrown away each time the liquid medical composition is ejected. On the other hand, the housing 2 side includes a battery 9 that supplies power to the igniter 71 included in the drive unit 7 of the syringe assembly 10. The power from the battery 9 is supplied between the electrode on the housing 2 side and the electrode on the drive unit 7 side of the syringe assembly 10 through wiring by the user pressing the button 8 provided on the housing 2. It should be noted that the shape and position of the electrode on the housing 2 side and the electrode on the drive unit 7 side of the syringe assembly 10 are designed so that the two electrodes are automatically in contact when the syringe assembly 10 is mounted on the housing 2. In addition, the housing 2 is a unit that can be used repeatedly as long as there is power remaining in the battery 9 that can be supplied to the drive unit 7. It should be noted that, in the casing 2 , when the power of the battery 9 is exhausted, only the battery 9 may be replaced and the casing 2 may continue to be used.

[0149] Next, the details of the syringe assembly 10 will be described. First, when describing the subassembly including the syringe barrel 3 and the plunger 4, the syringe barrel 3 has a receiving portion 32 formed therein as a space capable of receiving the liquid medical composition. Figure 1 As shown, the plunger 4 is configured to slide freely along the inner wall surface extending in the axial direction of the syringe barrel 3, and the inner wall surface of the syringe barrel 3 and the plunger 4 define a accommodating portion 32. In addition, the syringe barrel 3 has a nozzle portion 31 connected to the accommodating portion 32, and an ejection port 31a is formed on the top side of the nozzle portion 31. The nozzle portion 31 is a flow path whose cross-sectional area gradually decreases from the accommodating portion 32 side toward the ejection port 31a side, and is used to guide the liquid medical composition filled in the accommodating portion 32 to the ejection port 31a. Figure 1 In the example shown, the shape of the tip end side of the plunger 4 is substantially identical to the shape of the nozzle portion 31 .

[0150] Next, a subassembly including the syringe body 6, the piston 5 and the drive unit 7 will be described. The piston 5 is made of metal, for example, and is constructed to slide in a through hole formed inside the syringe body 6 by pressurizing the combustion product (combustion gas) generated by the igniter 71 of the drive unit 7. The syringe body 6 is a roughly cylindrical member that accommodates the piston 5, and the piston 5 is able to slide freely along the inner wall surface extending in the axial direction of the syringe body 6. It should be noted that the piston 5 can also be made of resin. In this case, metal can also be used in parts that require heat resistance and pressure resistance. In addition, if Figure 1 As shown, the piston 5 is integrally connected to the plunger 4 .

[0151] Next, the driving unit 7 will be described. Figure 1 As shown, the driving unit 7 is fixed to the base end side with the through hole in the syringe body 6 as a reference. The driving unit 7 has an igniter 71 as an electric igniter. The igniter 71 is configured in a manner to penetrate into the through hole in the syringe body 6, and ignition powder is contained therein. As the ignition powder, various gunpowders can be used as described above. In addition, the ignition powder can be contained in a gunpowder cup formed of an appropriate thin-walled metal, for example.

[0152] Next, the operation of the syringe 1 constructed as described above will be described. Figure 1 As shown, in a state where the syringe assembly 10 is mounted on the housing 2, the liquid medical composition is sucked from the ejection port 31a of the nozzle portion 31. Thus, the liquid medical composition can be filled into the housing portion 32. From this state, for example, when the ejection port 31a of the syringe 1 is brought into contact with the injection object, when the user presses the button 8 provided on the housing 2, this is used as a trigger to supply operating power from the battery 9 to the igniter 71 of the drive portion 7, and the igniter 71 operates. When the igniter 71 operates, the ignition powder is ignited and burns, generating combustion products (flame, combustion gas, etc.). As a result, for example, the gunpowder cup of the igniter 71 is cracked, and the combustion gas of the ignition powder is released into the through hole of the syringe body 6. As a result, the pressure in the through hole of the syringe body 6 rises sharply, and the piston 5 is pressed toward the top side of the syringe body 6, and as a result, the piston 5 slides toward the top side along the inner wall surface of the through hole of the syringe body 6. As described above, since the plunger 4 is integrally connected to the piston 5, the plunger 4 also slides along the inner wall surface of the syringe barrel 3 in a manner linked with the piston 5. That is, by pressing the plunger 4 into the nozzle portion 31 located at the top end side of the syringe barrel 3, the volume of the container 32 containing the liquid medical composition is reduced, and the pressure is increased rapidly. As a result, the liquid medical composition filled in the container 32 is pressed into the nozzle portion 31 and ejected from the ejection port 31a at high pressure. Thus, the liquid medical composition can be injected into the injection object.

[0153] In addition, Figure 1 Although no additional explosive component is particularly arranged in the syringe body 6 shown, in order to adjust the pressure shift applied to the liquid medical composition via the piston 5, a gas generating agent that generates gas by burning the combustion product generated by the combustion of the explosive in the igniter 71 may be arranged in the igniter 71 and in the through hole of the syringe body 6. The configuration of arranging the gas generating agent in the igniter 71 is a well-known technology as disclosed in International Publication No. 01-031282 and Japanese Patent Application Publication No. 2003-25950. In addition, as an example of the gas generating agent, a single-base smokeless powder containing 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 generating agents used for gas generators for air bags and gas generators for seat belt pretensioners can also be used. By adjusting the size, shape, and especially the surface shape of the gas generating agent when it is arranged in the through hole, the combustion completion time of the gas generating agent can be changed, thereby, the pressure shift applied to the liquid medical composition can be set to the desired shift, that is, the shift that can make the liquid medical composition properly reach the injection object. In this embodiment, the driving part 7 also includes a gas generating agent used as needed. In this embodiment, the "pressurizing part" includes a plunger 4 and a piston 5.

[0154] Another embodiment of the present disclosure is an injector that injects a liquid pharmaceutical composition into an injection subject without using an injection needle.

[0155] The injector has:

[0156] a containing portion containing a liquid pharmaceutical composition;

[0157] a nozzle portion, connected to the containing portion, the nozzle portion having an ejection port for ejecting the liquid medical composition toward an injection object; and

[0158] a pressurizing unit that, when in operation, pressurizes the liquid medical composition contained in the containing unit, thereby ejecting the liquid medical composition from the ejection port toward the injection target,

[0159] The liquid pharmaceutical composition contains a biological functional substance and has an osmotic pressure greater than 400 mOsmol / kg and less than 1000 mOsmol / kg.

[0160] The injector of this embodiment is an injector described in the above embodiment that injects a liquid pharmaceutical composition into an injection target without using an injection needle, wherein the liquid pharmaceutical composition of the above embodiment is contained in the container. The description of this embodiment refers to the description of the above embodiment.

[0161] Another embodiment of the present disclosure is a method for injecting a liquid pharmaceutical composition into an injection object, comprising: injecting the liquid pharmaceutical composition into the injection object using an injector that injects the liquid pharmaceutical composition into the injection object without using an injection needle, wherein the liquid pharmaceutical composition contains a functional substance in vivo and has an osmotic pressure greater than 400mOsmol / kg and less than 1000mOsmol / kg.

[0162] The method of this embodiment is a method of injecting the liquid pharmaceutical composition of the embodiment into a subject using an injector for injecting the liquid pharmaceutical composition into a subject without using an injection needle. The description of this embodiment refers to the description of the above embodiment.

[0163] When a liquid pharmaceutical composition is injected into a subject by the method of this embodiment, as described in the embodiment, the physiological activity of the in vivo functional substance contained in the liquid pharmaceutical composition is exerted in the subject.

[0164] Therefore, the method of this embodiment can be as follows: it includes the step of injecting a liquid medicinal composition containing a functional substance in the body and having an osmotic pressure greater than 400mOsmol / kg and less than 1000mOsmol / kg into the injection object using an injector that injects the liquid medicinal composition into the injection object without passing through an injection needle, and the method enables the physiological activity of the functional substance in the body to be exerted in the injection object.

[0165] Example

[0166] Although Examples are described below, all Examples are not to be construed as Examples in a limiting sense.

[0167] The following animal experiments were conducted at the Animal Experimentation Facility of the School of Medicine, Osaka University, in accordance with the Osaka University Animal Experimentation Regulations established by the Osaka University Animal Experimentation Committee.

[0168] In addition, the osmotic pressure of the liquid pharmaceutical composition used in the following test examples was measured using Osmometer 3000 basic manufactured by Gonotec.

[0169] [Test Example 1]

[0170] In this test example, plasmid DNA containing the GFP gene was used as the in vivo functional substance, and phosphate buffered saline (PBS(-)) was used as the pharmacologically acceptable reagent (solvent). Figure 1The injector described in and having a nozzle diameter of 0.1 mm was used. 25 mg of ZPP was used as the ignition powder, and 40 mg of a single-base smokeless powder (containing 98% by mass of nitrocellulose, 0.8% by mass of diphenylamine, and 1.2% by mass of potassium sulfate) was used as the gas generating agent.

[0171] (Preparation of Liquid Pharmaceutical Composition)

[0172] PBS-tablet (manufactured by TAKARA BIO) (PBS (-)) was dissolved in water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.) to prepare 10 times PBS. Using the water for injection, the prepared 10 times PBS was diluted to twice the concentration when injected into 10-week-old male balb / c mice (CLEA Japan) as test animals. Then, it was mixed with CMV-DASHER-GFP (manufactured by ATUM) (solvent: TE) (1.0 mg / mL), which is a plasmid DNA containing the GFP gene, in a ratio of 1:1 to prepare a liquid pharmaceutical composition. That is, the final concentration of the plasmid DNA containing the GFP gene was 0.5 mg / mL.

[0173] Then, a liquid pharmaceutical composition is prepared so that the final osmotic pressure (the final ratio of PBS is indicated in parentheses) is 198.00 mOsmol / kg (0.5 times PBS), 346.00 mOsmol / kg (1 times PBS), 624.00 mOsmol / kg (2 times PBS), 899.00 mOsmol / kg (3 times PBS), 1154.00 mOsmol / kg (4 times PBS), or 1424.00 mOsmol / kg (5 times PBS).

[0174] (Injection into 10-week-old male balb / c mice)

[0175] After inhalation anesthesia with isoflurane was performed on 10-week-old male balb / c mice, hair on the back was trimmed using an animal electric clipper. Then, the back was disinfected with alcohol, and the liquid pharmaceutical composition was injected into multiple sites on the back using the injector. 20 μL of the liquid pharmaceutical composition was injected into each site.

[0176] (Evaluation of Gene Expression)

[0177] One day after the injection, the mouse was euthanized. Then, a skin piece of a thickness that can be observed under a microscope was collected from the back, attached to a Matsunami glass-bottomed culture dish, and observed using a fluorescence microscope BZ-X710 (manufactured by KEYENCE). A bright field image (exposure time: 1 / 350 s) and a fluorescent image of the GFP channel (exposure time: 1 / 20 s) were obtained, and a superimposed image (bright field image + fluorescent image) was obtained as needed.

[0178] (Analysis of Gene Expression)

[0179] The fluorescent image was analyzed using an analysis application (Hybrid Cell Counter) (manufactured by KEYENCE Co., Ltd.) The analysis was performed under the conditions where the brightness was set to 10 and the area designation was off.

[0180] [Comparative Test Example 1]

[0181] The same procedure as in Test Example 1 was carried out except that a syringe with a 30G needle (Dentronics Needle 30G manufactured by Misawa Medical Industry Co., Ltd.) was used as the injector.

[0182] It should be noted that the following cases were also tested: no treatment (no injection of the liquid pharmaceutical composition); Figure 1 The injector described in is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition; the needle syringe with a 30G needle (Dentronics needle 30G manufactured by MISAWA MEDICAL INDUSTRY Co., Ltd.) is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition.

[0183] The results are shown in Table 1 and Figure 2 It should be noted that the "Relative Brightness (1)" in the table is a value obtained by converting the brightness when 57.00 mOsmol / kg (TE) is used as 1.00, and the "Relative Brightness (2)" is a value obtained by converting the brightness when 346.00 mOsmol / kg (1×PBS) is used as 1.00.

[0184] Comparison between the conditions of Test Example 1 suggests that, for example, the luminosity when the osmotic pressure is greater than 400 mOsmol / kg and 1000 mOsmol / kg or less is greater than the luminosity when the osmotic pressure is 400 mOsmol / kg.

[0185] In addition, the luminance of Test Example 1 was greater than that of Comparative Test Example 1 at any osmotic pressure. From the results of relative luminance (2) (sometimes referred to as the gene expression increase rate), it can be seen that the luminance of Comparative Test Example 1 was greater than that of Experimental Example 1. Figure 1In the case of the injector described in, and in the case of osmotic pressure of 624.00mOsmol / kg (2 times PBS) and 899.00mOsmol / kg (3 times PBS), the value is more than 1 times. Furthermore, even in the comparison with the relative luminosity (2) (sometimes referred to as the gene expression increase rate) of Comparative Test Example 1 corresponding to these ranges, a larger value is shown in Test Example 1. From these results, it can be seen that the gene expression enhancement effect can be obtained as the osmotic pressure increases.

[0186] It should be noted that, interestingly, in Comparative Test Example 1, the result that the gene expression increase rate due to osmotic pressure exceeded 1 was also confirmed. This result suggests that even when a syringe with a needle is used, an increase in osmotic pressure is related to the result of increased gene expression. However, the gene expression increase rate in Test Example 1 showed a larger value than that in the case of Comparative Test Example 1. This shows that the gene expression enhancement effect due to the increase in osmotic pressure is greater when using Figure 1 This is particularly true in the case of the injector described in .

[0187] [Table 1]

[0188]

[0189] [Test Example 2]

[0190] In this test example, the same procedure as in Test Example 1 was performed except that sodium chloride (NaCl) dissolved in water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.) was used as a pharmacologically acceptable agent (solvent) and 11-week-old male balb / c mice (CLEA Japan) were used as test animals.

[0191] [Comparative Test Example 2]

[0192] The same procedure as in Test Example 2 was carried out except that a syringe with a 30G needle (Dentronics Needle 30G manufactured by Misawa Medical Industry Co., Ltd.) was used as the injector.

[0193] It should be noted that the following situations were also tested: Figure 1 The injector described in is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition; the needle syringe with a 30G needle (Dentronics needle 30G manufactured by MISAWA MEDICAL INDUSTRY Co., Ltd.) is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition.

[0194] The results are shown in Table 2 and Figure 3 It should be noted that the concentration in the top column of the table is the concentration of NaCl in the liquid pharmaceutical composition, "Relative luminosity (1)" is the value obtained by converting the luminosity when 52.00 mOsmol / kg (TE) is set to 1.00, and "Relative luminosity (2)" is the value obtained by converting the luminosity when 324.00 mOsmol / kg (150 mM) is set to 1.00.

[0195] Comparison between the conditions of Test Example 2 suggests that, for example, the luminosity when the osmotic pressure is greater than 400 mOsmol / kg and 1000 mOsmol / kg or less is greater than the luminosity when the osmotic pressure is 400 mOsmol / kg.

[0196] In addition, the luminance of Test Example 2 was greater than that of Comparative Test Example 2 regardless of the osmotic pressure. Furthermore, from the results of relative luminance (2) (sometimes referred to as the gene expression increase rate), when using Figure 1 In the case of the injector described in , and when the osmotic pressure is 460.00mOsmol / kg (225mM), 605.00mOsmol / kg (300mM), and 879.00mOsmol / kg (450mM), it is a value exceeding 1 times. Therefore, it can be seen that the gene expression enhancement effect obtained with the increase of osmotic pressure is not dependent on the type of salt used.

[0197] [Table 2]

[0198]

[0199] [Test Example 3]

[0200] In this Test Example, the same procedure as in Test Example 1 was performed except that glucose dissolved in water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.) was used as a pharmacologically acceptable agent (solvent) and 11-week-old male balb / c mice (CLEA Japan) were used as test animals.

[0201] [Comparative Test Example 3]

[0202] The same procedure as in Test Example 3 was carried out except that a syringe with a 30G needle (Dentronics Needle 30G manufactured by Misawa Medical Industry Co., Ltd.) was used as the injector.

[0203] It should be noted that the following situations were also tested: Figure 1The injector described in is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition; the needle syringe with a 30G needle (Dentronics needle 30G manufactured by MISAWA MEDICAL INDUSTRY Co., Ltd.) is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition.

[0204] The results are shown in Table 3 and Figure 4 It should be noted that the concentration in the top column of the table is the concentration of glucose in the liquid pharmaceutical composition, and the "Relative Brightness (1)" is the value obtained by converting the brightness when 53.00 mOsmol / kg (TE) is set to 1.00, and the "Relative Brightness (2)" is the value obtained by converting the brightness when 312.00 mOsmol / kg (54 mg / mL) is set to 1.00.

[0205] Comparison between the conditions of Test Example 3 suggests that, for example, the luminosity when the osmotic pressure is greater than 400 mOsmol / kg and 1000 mOsmol / kg or less is greater than the luminosity when the osmotic pressure is 400 mOsmol / kg.

[0206] In addition, the luminance of Test Example 3 was greater than that of Comparative Test Example 3 regardless of the osmotic pressure. Furthermore, from the results of relative luminance (2) (sometimes referred to as the gene expression increase rate), when using Figure 1 In the case of the injector described in , and when the osmotic pressure is 436.00mOsmol / kg (81mg / mL), 584.00mOsmol / kg (108mg / mL), 872.00mOsmol / kg (162mg / mL), it is a value exceeding 1 times. Therefore, it is shown that the gene expression enhancement effect obtained with the increase of osmotic pressure is not dependent on the type of solute used.

[0207] [Table 3]

[0208]

[0209] [Test Example 4]

[0210] In this test example, the same procedure as in Test Example 1 was performed except that a spring-loaded needleless syringe (BEAUTTO, hyaluronic acid syringe, Amazon Standard Identification Number (ASIN): B08NCHTRHZ) was used as an injector and 11-week-old male balb / c mice (CLEA Japan) were used as test animals.

[0211] [Comparative Test Example 4]

[0212] The same procedure as in Test Example 4 was carried out except that a syringe with a 30G needle (Dentronics Needle 30G manufactured by MISAWA MEDICAL INDUSTRY CO., LTD.) was used as the injector.

[0213] It should be noted that the following cases were also tested: a case without treatment (a case where the liquid pharmaceutical composition was not injected); a case where a spring-type needleless syringe (BEAUTTO, hyaluronic acid syringe, Amazon Standard Identification Number (ASIN): B08NCHTRHZ) was used as an injector, and TE (Tris-EDTA, pH: 8.0) was used as the liquid pharmaceutical composition; a case where a needle syringe with a 30G needle (Dentronics needle 30G manufactured by MISAWA MEDICAL INDUSTRY Co., Ltd.) was used as an injector, and TE (Tris-EDTA, pH: 8.0) was used as the liquid pharmaceutical composition.

[0214] The results are shown in Table 4 and Figure 5 It should be noted that the "Relative Brightness (1)" in the table is a value obtained by converting the brightness when 57.00 mOsmol / kg (TE) is used as 1.00, and the "Relative Brightness (2)" is a value obtained by converting the brightness when 346.00 mOsmol / kg (1×PBS) is used as 1.00.

[0215] When the conditions of Test Example 4 are compared with each other, it is suggested that there is a high possibility that the same tendency as that of Test Example 1 is exhibited.

[0216] In addition, since the optimal conditions for the injection of the spring-type needleless syringe were not studied, the gene expression was kept at a low level. However, the results of relative brightness (2) (sometimes referred to as the gene expression increase rate) showed that the value exceeded 1 times when the osmotic pressure was 624.00 mOsmol / kg (2 times PBS). Therefore, it was shown that the gene expression enhancement effect with the increase of osmotic pressure is not limited to the injector that uses the combustion energy of the ignition powder as the injection energy, but can also be obtained with the spring-type needleless syringe.

[0217] [Table 4]

[0218] Table 4

[0219]

[0220]

[0221] [Test Example 5]

[0222] In this experimental example, mRNA (unmodified) (manufactured by OZ Co., Ltd., Funakoshi Cat#.MRNA15-100) (solvent: TE) (0.2 mg / mL) encoding GFP as a reporter protein was used as the in vivo functional substance, 11-week-old male balb / c mice (CLEA Japan) were used as test animals, and the final concentration of mRNA was set to 0.1 mg / mL (2.0 μg per injection). Except for this, the same procedure as in Experiment 1 was performed.

[0223] [Comparative Test Example 5]

[0224] The same procedure as in Test Example 5 was carried out except that a syringe with a 30G needle (Dentronics Needle 30G manufactured by MISAWA MEDICAL INDUSTRY CO., LTD.) was used as the injector.

[0225] It should be noted that the following cases were also tested: no treatment (no injection of the liquid pharmaceutical composition); Figure 1 The injector described in is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition; the needle syringe with a 30G needle (Dentronics needle 30G manufactured by MISAWA MEDICAL INDUSTRY Co., Ltd.) is used as the injector, and TE (Tris-EDTA, pH: 8.0) is used as the liquid pharmaceutical composition.

[0226] The results are shown in Table 5 and Figure 6 It should be noted that the "Relative Brightness (1)" in the table is a value obtained by converting the brightness when 57.00 mOsmol / kg (TE) is used as 1.00, and the "Relative Brightness (2)" is a value obtained by converting the brightness when 346.00 mOsmol / kg (1×PBS) is used as 1.00.

[0227] When the conditions of Test Example 5 are compared with each other, it is suggested that there is a high possibility that the same tendency as that of Test Example 1 is exhibited.

[0228] In addition, from the results of relative luminosity (2) (sometimes called the rate of increase in gene expression), Figure 1In the case of the described injector, when the osmotic pressure was 624.00 mOsmol / kg (2 times PBS), the value was more than 1 times. Therefore, it was shown that the gene expression enhancement effect accompanying the increase of osmotic pressure can be obtained by nucleic acids in a broad sense including mRNA.

[0229] [Table 5]

[0230] Table 5

[0231]

[0232] Description of Reference Numerals

[0233] 1: syringe; 2: housing; 3: syringe barrel; 4: plunger; 5: piston; 6: syringe body; 7: drive unit; 8: button; 9: battery; 10: syringe assembly; 31: nozzle unit; 31a: ejection port; 32: storage unit; 71: igniter.

Claims

1. A liquid pharmaceutical composition, wherein: The liquid pharmaceutical composition contains a functional substance in vivo, The osmotic pressure of the liquid pharmaceutical composition is greater than 400 mOsmol / kg and less than 1000 mOsmol / kg, The liquid medical composition is injected into the injection target using an injector that injects the liquid medical composition into the injection target without using an injection needle.

2. The liquid pharmaceutical composition according to claim 1, wherein The liquid pharmaceutical composition comprises a pharmacologically acceptable agent.

3. The liquid pharmaceutical composition according to claim 2, wherein The pharmacologically acceptable agent is an ionic agent.

4. The liquid pharmaceutical composition according to claim 3, wherein The ionic reagent is a buffer using phosphoric acid.

5. The liquid pharmaceutical composition according to claim 4, wherein The buffer using phosphate is phosphate buffered saline.

6. The liquid pharmaceutical composition according to claim 3, wherein The ionic reagent is sodium chloride.

7. The liquid pharmaceutical composition according to claim 2, wherein The pharmacologically acceptable agent is a non-ionic agent.

8. The liquid pharmaceutical composition according to claim 7, wherein The non-ionic reagent is glucose.

9. The liquid pharmaceutical composition according to claim 1 or 2, wherein The liquid pharmaceutical composition is injected into the injection target by using an injector that injects the liquid pharmaceutical composition into the injection target without using an injection needle, and is injected into the injection target by jet injection.

10. The liquid pharmaceutical composition according to claim 1 or 2, wherein The functional substance in the organism is nucleic acid.

11. An injector for injecting a liquid pharmaceutical composition into an injection object without using an injection needle, The injector has: a containing portion containing a liquid pharmaceutical composition; a nozzle portion, connected to the containing portion, the nozzle portion having an ejection port for ejecting the liquid medical composition toward an injection object; and a pressurizing unit that, when in operation, pressurizes the liquid medical composition contained in the containing unit, thereby ejecting the liquid medical composition from the ejection port toward the injection target, The liquid pharmaceutical composition contains a biological functional substance and has an osmotic pressure greater than 400 mOsmol / kg and less than 1000 mOsmol / kg.

12. A method for injecting a liquid pharmaceutical composition into an injection subject, wherein: include: A liquid pharmaceutical composition is injected into a subject using an injector that injects the liquid pharmaceutical composition into the subject without using an injection needle. The liquid pharmaceutical composition contains a biological functional substance and has an osmotic pressure of more than 400 mOsmol / kg and less than 1000 mOsmol / kg.

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