Pharmaceutical composition and particles

Abstract

The present invention provides a pharmaceutical composition which contains a population of cells, wherein the cells comprise cells that accumulate in an inflamed site and particles that are introduced into the cells and contain a bioabsorbable polymer and a drug, and the drug is cyclosporin or minoxidil.

Description

Pharmaceutical compositions and particles

[0001] The present invention relates to a pharmaceutical composition, a particle, and the like.

[0002] When using small molecular weight compounds as pharmaceuticals, their toxicity must be evaluated. If the toxic concentration of a candidate compound is close to its effective concentration, it is difficult to use the compound as a pharmaceutical. For this reason, only a very limited number of small molecular weight compounds are used as pharmaceuticals.

[0003] Recently, cell preparations have been used as pharmaceuticals. For example, Patent Document 1 discloses a preparation containing statin-encapsulated nanoparticles in which a statin is encapsulated in nanoparticles containing a bioabsorbable polymer, the nanoparticles having a number-average particle size of less than 1000 nm, used to enhance stem cell function, as well as stem cells that have taken up and contained the statin-encapsulated nanoparticles. Furthermore, Patent Documents 2 and 3 disclose enhancing stem cell function by incorporating statin-encapsulated nanoparticles in stem cells. Non-Patent Document 1 discloses PLGA-PEG nanoparticles loaded with docetaxel.

[0004] International Publication No. 2016 / 076227 International Publication No. 2017 / 191808 Japanese Patent Application Publication No. 2019-196342 J Thoracic Oncol., Vol.13No.10S, 2018, p.S455-S456

[0005] Patent Documents 1 to 3 all merely disclose formulations used to enhance stem cell function. However, if the effects of drugs such as low-molecular-weight compounds can be exerted at lower concentrations than usual, even drugs whose toxicity and effective concentrations are close to each other may be usable as pharmaceuticals. For this reason, there is a demand for drug administration methods that can reduce the effective concentration of drugs.

[0006] However, the administration method of such drugs and the pharmaceutical composition to be used in said administration method have not yet been established.

[0007] Therefore, an object of the present invention is to provide a pharmaceutical composition that can reduce the dosage of the drug per dosage unit compared to when the drug is used alone, and particles, etc. used in the pharmaceutical composition.

[0008] As a result of intensive research to solve the above-mentioned problems, the inventors discovered that the above-mentioned problems can be solved by using a population of cells into which particles containing a bioabsorbable polymer and a drug have been introduced into cells that accumulate at the site of inflammation, and thus completed the present invention.

[0009] The present invention includes the following embodiments. [1] A pharmaceutical composition comprising a population of cells, wherein the cells are cells that accumulate at an inflammatory site and into which particles containing a bioabsorbable polymer and a drug have been introduced, and the drug is cyclosporine or minoxidil. [2] The pharmaceutical composition according to [1], for use in a method comprising administering cyclosporine or minoxidil at a dose less than the dose per dosage unit of cyclosporine or minoxidil. [3] The pharmaceutical composition according to [1] or [2], wherein the cells comprise at least one of immune cells and mesenchymal stem cells. [4] The pharmaceutical composition according to [1] or [2], for suppressing an immune response, wherein the cells comprise peripheral blood mononuclear cells and the drug is cyclosporine. [5] The pharmaceutical composition according to [1] or [2], wherein the concentration of cyclosporine contained in the cells is 1 ng / 10 ５ [6] The pharmaceutical composition according to claim 4, wherein the cells are mesenchymal stem cells or more. [6] The pharmaceutical composition according to [1] or [2], for promoting angiogenesis, wherein the cells comprise mesenchymal stem cells and the drug is minoxidil. [7] The pharmaceutical composition according to claim 4, wherein the cells comprise mesenchymal stem cells or more ...8] The pharmaceutical composition according to claim 4, wherein the cells comprise mesenchymal stem cells and the drug is minoxidil. [9] The pharmaceutical composition according to claim 4, wherein the cells comprise mesenchymal stem cells or more. ５The pharmaceutical composition according to claim 6, which is a cell or more. [8] A pharmaceutical composition for promoting angiogenesis, comprising a population of mesenchymal stem cells and minoxidil. [9] A pharmaceutical composition comprising a population of cells, wherein the cells are cells that accumulate at an inflammatory site, into which particles comprising a bioabsorbable polymer and a drug have been introduced, for use in a method comprising administering the drug at a dose smaller than the dose per unit of the drug.

[10] Particles for use in a method comprising administering the drug at a dose smaller than the dose per unit of the drug, the particles comprising a bioabsorbable polymer and the drug, and being introduced into cells that accumulate at an inflammatory site.

[11] A method for administering a drug, comprising: encapsulating a drug in a bioabsorbable polymer to prepare drug-encapsulated particles; introducing the drug-encapsulated particles into cells that accumulate at an inflammatory site; and administering the cells.

[12] The method according to

[11] , comprising administering the drug at a dose less than the dose per dosage unit of the drug.

[13] The method according to

[11] or

[12] , wherein the cells accumulate at an inflammatory site.

[14] The method according to any one of

[11] to

[13] , wherein the drug is cyclosporine or minoxidil.

[15] The method according to any one of

[11] to

[14] , wherein the cells comprise at least one of immune cells and mesenchymal stem cells.

[16] The method according to any one of

[11] to

[15] , wherein the cells comprise peripheral blood mononuclear cells, and the drug is cyclosporine.

[17] The method according to any one of

[11] to

[15] , wherein the cells comprise mesenchymal stem cells, and the drug is minoxidil.

[0010] The present invention may include the following. [A1] A method for treating or preventing a disease, comprising administering to a subject in need thereof the pharmaceutical composition of any one of [1] to

[10] . [B1] The pharmaceutical composition of any one of [1] to

[10] for use in suppressing an immune response or promoting angiogenesis. [C1] Use of the pharmaceutical composition of any one of [1] to

[10] in the manufacture of a medicament for suppressing an immune response or promoting angiogenesis. [D1] Use of the pharmaceutical composition of any one of [1] to

[10] for suppressing an immune response or promoting angiogenesis. In [A1] to [D1], the pharmaceutical composition of any one of [1] to

[10] may be a population of cells of any one of [1] to

[10] . [A2] A method for treating or preventing a disease, comprising administering to a subject in need thereof the population of cells of any one of [1] to

[10] . [B3] The population of cells of any one of [1] to

[10] for use in suppressing an immune response or promoting angiogenesis. [C4] The population of cells according to any one of [1] to

[10] in the manufacture of a medicament for suppressing an immune response or promoting angiogenesis. [D2] Use of the population of cells according to any one of [1] to

[10] for suppressing an immune response or promoting angiogenesis.

[0011] According to the present invention, it is possible to provide a pharmaceutical composition that can reduce the dosage of a drug per dosage unit compared to when the drug is used alone, and particles and the like used in the pharmaceutical composition.

[0012] Results of immune response in Example 1. Results of observation under a fluorescence microscope under each condition in Example 2. Arrows indicate blood vessel-like structures. Note that in each figure, mcM means μM, and hAP means that mesenchymal stem cells into which minoxidil-encapsulated PLGA particles were introduced were used.

[0013] Hereinafter, an embodiment of the present invention (hereinafter referred to as "the present embodiment") will be described in detail. However, the present invention is not limited to the following embodiment, and various modifications are possible without departing from the gist of the present invention.

[0014] [Pharmaceutical Composition] In one aspect of this embodiment, the pharmaceutical composition comprises a population of cells, wherein the cells are cells that accumulate at an inflammatory site and into which particles comprising a bioabsorbable polymer and a drug have been introduced. Examples of the drug include, but are not limited to, cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, and irinotecan. In another aspect of this embodiment, the pharmaceutical composition comprises a population of cells, wherein the cells are cells that accumulate at an inflammatory site and into which particles comprising a bioabsorbable polymer and a drug have been introduced, and the pharmaceutical composition is for use in a method comprising administering the drug at a dose lower than the dose per dosage unit of the drug. In yet another aspect of this embodiment, the pharmaceutical composition may be a population of cells that accumulate at an inflammatory site and into which particles comprising a bioabsorbable polymer and a drug have been introduced. Furthermore, the population of cells may be a population of cells that accumulate at an inflammatory site. In this specification, a description of a pharmaceutical composition comprising a population of cells may be understood as a description of a population of cells.

[0015] As will be shown in the examples below, in this embodiment, a drug is encapsulated in particles containing a bioabsorbable polymer, the particles are introduced into cells that accumulate at the site of inflammation, and the drug is administered using the cells, thereby achieving the effects of the drug with a smaller dose than when the drug is administered alone.

[0016] This is presumably because, with such a pharmaceutical composition, the cells accumulate at the affected area due to their ability to accumulate at the site of inflammation, and the bioabsorbable polymer of the particles contained in the accumulated cells decomposes, resulting in the sustained release of the drug, thereby simultaneously exerting the effects of the cells and the drug, although this presumption is not intended to limit the scope of the present invention.

[0017] (Cells) The pharmaceutical composition of this embodiment includes cells that accumulate at inflammatory sites and into which particles containing a bioabsorbable polymer and a drug have been introduced (hereinafter also referred to as "cells of this embodiment"). Note that the pharmaceutical composition of this embodiment may also include cells other than the cells of this embodiment. Examples of such cells include cells that have not been introduced with particles containing a bioabsorbable polymer and a drug and that accumulate at inflammatory sites, and cells that have been introduced with particles containing a bioabsorbable polymer and a drug and that do not accumulate at inflammatory sites.

[0018] The cells that accumulate at the inflammatory site are not particularly limited as long as they have the ability to accumulate at the inflammatory site, and examples thereof include immune cells and mesenchymal stem cells.

[0019] The immune cells are not particularly limited as long as they are cells involved in immune responses, and may be, for example, T cells, B cells, natural killer (NK) cells, macrophages, monocytes, dendritic cells, and granulocytes (eosinophils, neutrophils, basophils, mast cells, etc.), or a combination of one or more of these. The immune cells may also be a combination of multiple types of cells such as mononuclear cells, particularly peripheral blood mononuclear cells.

[0020] The tissue from which mesenchymal stem cells are derived is not particularly limited, as long as they are somatic stem cells derived from mesenchyme and have the ability to self-renew and differentiate. Examples of mesenchymal stem cells include mesenchymal stem cells derived from adipose tissue, bone marrow, dental pulp, blood (peripheral blood, umbilical cord blood, etc.), placenta, umbilical cord, synovium, periosteum, perichondrium, muscle, ligament, tendon, meniscus, or skin. Among these, adipose-derived mesenchymal stem cells are preferably used. Adipose-derived mesenchymal stem cells can be obtained from adipose tissue, which can be easily obtained from subcutaneous fat using minimally invasive techniques such as liposuction. Adipose-derived mesenchymal stem cells can be abundantly collected by extraction and separation from the adipose tissue obtained as described above using a Celution System (Cytori, Inc.) or the like.

[0021] The cells that accumulate at the inflammatory site may be derived from a non-human animal or a human, preferably a human. Non-human animals include, but are not limited to, mice, rats, guinea pigs, hamsters, rabbits, monkeys, cows, minipigs, pigs, sheep, goats, dogs, and cats.

[0022] Furthermore, the cells that accumulate at the site of inflammation may be derived from cells of the individual to whom the pharmaceutical composition of this embodiment or the cells of this embodiment is to be administered (autologous cells), or may be derived from cells other than that individual (allogeneic cells). Furthermore, the cells that accumulate at the site of inflammation may be xenogeneic or allogeneic cells.

[0023] The cells that accumulate at the site of inflammation may be cells collected from a living body or cells obtained by culturing such cells, or may be cells obtained by inducing differentiation from pluripotent stem cells, such as embryonic stem cells (ES cells) and iPS cells.

[0024] The method for introducing particles containing a bioabsorbable polymer and a drug (hereinafter also referred to as "particles of this embodiment" in this specification) into cells is not particularly limited, but includes a method of co-culturing the particles of this embodiment with cells. By co-culturing the particles of this embodiment with cells that accumulate at the inflammatory site, the cells take up the particles of this embodiment into the cells by endocytosis or membrane permeation, and the cells of this embodiment can be easily obtained.

[0025] The conditions for co-culturing the particles of this embodiment with cells that accumulate at inflammatory sites are not particularly limited, and may be, for example, normal culture conditions for culturing cells that accumulate at inflammatory sites or culture in an appropriate buffer. The culture time is also not particularly limited, but from the viewpoint of ensuring that the cells take up the particles of this embodiment, it is preferably 5 minutes to 2 hours, 10 minutes to 1 hour, or 15 to 45 minutes. After co-culturing the particles of this embodiment with the cells, the cell population may be washed as appropriate to remove particles of this embodiment that have not been taken up by the cells.

[0026] The pharmaceutical composition of this embodiment may be in the form of a cell suspension containing the cells of this embodiment, or in the form of a cell sheet or sheet-like cell culture obtained by culturing the cells of this embodiment to form a sheet-like tissue. The cell sheet may be a composite cell sheet having a layer structure of the cells of this embodiment and feeder cells.

[0027] (Particles) The particles of this embodiment are particles containing a bioabsorbable polymer and a drug, and can also be described as drug-encapsulated particles in which a drug is encapsulated in a particle containing a bioabsorbable polymer.

[0028] The bioabsorbable polymer is not particularly limited as long as it is a bioabsorbable or biodegradable polymeric material, and examples thereof include polymers and copolymers of hydroxy acids such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and poly(lactic-co-glycolic acid) (PLGA); polymers and copolymers such as PEG, polyanhydrides, poly(ortho)esters, polyesters, polyurethanes, poly(butyric acid), poly(valeric acid), poly(caprolactone), poly(hydroxyalkanoic acid), and poly(lactide-co-caprolactone); and natural polymers such as proteins such as albumin, collagen, gelatin, and prolamins, alginates, cellulose derivatives, and polyhydroxyalkanoates. Among these, preferred bioabsorbable polymers are polymers and copolymers of hydroxy acids such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and poly(lactic-co-glycolic acid) (PLGA), with poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and poly(lactic-co-glycolic acid) (PLGA) being more preferred. The bioabsorbable polymer preferably includes at least one selected from the group consisting of poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and poly(lactic-co-glycolic acid) (PLGA). The term "bioabsorbable polymer" may also be referred to as "biodegradable polymer."

[0029] The particles of this embodiment may contain components other than the bioabsorbable polymer and the drug. Such components include water, distilled water for injection, saline, glucose solution, isotonic solution (e.g., solutions of sodium chloride, potassium chloride, glycerin, mannitol, sorbitol, boric acid, borax, propylene glycol, etc.), other aqueous solvents, and aqueous or oily bases. The drug contained in the particles of this embodiment will be described later.

[0030] The particle size of the particles of this embodiment is not particularly limited as long as it is within a range that can be taken up by cells, and may be changed appropriately depending on the type of cell. The particle size of the particles of this embodiment may be, for example, 1000 nm or less, preferably 50 nm to 400 nm, and more preferably 100 nm to 200 nm. Therefore, the particles of this embodiment can also be called nanoparticles. The particle size of the particles of this embodiment can be measured by a light scattering method using a particle suspension.

[0031] The method for forming the particles of this embodiment is not particularly limited, but may be, for example, by encapsulating other components, including a drug, in a bioabsorbable polymer. Specifically, the particles may be formed by, for example, spherical crystallization. The spherical crystallization method is a method that allows spherical crystal particles to be designed and their physical properties directly controlled and processed by controlling the crystal generation and growth process in the final process of compound synthesis. Examples of spherical crystallization methods include emulsion solvent diffusion (ESD).

[0032] The emulsion solvent diffusion method can be carried out, for example, by the following method: A bioabsorbable polymer, a drug, and optionally other components are dissolved in a good solvent that can dissolve the bioabsorbable polymer to obtain a mixed solution. The mixed solution is then added dropwise to a poor solvent that does not dissolve the bioabsorbable polymer while stirring, thereby forming polymer nanoparticles in the form of spherical crystalline particles.

[0033] (Drug) In the pharmaceutical composition of this embodiment, the drug is contained in the particles introduced into the cells. Therefore, in the pharmaceutical composition of this embodiment, the drug is mainly contained in the particles within the cells, but may also be contained in other forms. For example, in the pharmaceutical composition of this embodiment, the drug may be contained in the region outside the particles within the cells, or may be contained in the region outside the cells.

[0034] The drug encapsulated in the particles of this embodiment is not particularly limited and can be selected depending on the intended use of the pharmaceutical composition of this embodiment. The drug encapsulated in the particles of this embodiment may be, for example, a lipophilic drug. When a lipophilic drug is used, after the drug is released from the particles of this embodiment, the drug can suitably permeate the cell membrane of the cells containing the particles of this embodiment and the cell membranes of other cells in the affected area, which tends to more effectively and reliably achieve the effects of this embodiment. In one aspect of the pharmaceutical composition of this embodiment, the drug is cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, irinotecan, or the like. The drug may be cyclosporine, minoxidil, paclitaxel, cisplatin, or irinotecan, or may be cyclosporine, minoxidil, cisplatin, or irinotecan, or may be cyclosporine or minoxidil.

[0035] The amount of drug encapsulated per particle of the present embodiment is not particularly limited and can be adjusted appropriately depending on the type of drug and the type of cells into which the particles of the present embodiment are introduced. The amount of drug encapsulated per particle can be controlled by adjusting the conditions in the above-mentioned method for forming particles of the present embodiment. For example, when forming particles of the present embodiment using an emulsion solvent diffusion method, the amount of drug encapsulated per particle can be controlled by adjusting the amount of drug added together with the bioabsorbable polymer, adjusting the stirring speed of the poor solvent, or adjusting the time from when the mixed solution containing the bioabsorbable polymer and drug is dropped into the poor solvent until the particles are removed.

[0036] The concentration of the drug encapsulated in the particles of this embodiment may be adjusted appropriately depending on the type of drug, the mixing ratio of the particles of this embodiment to the cells, etc. For example, the drug concentration in the suspension of the particles of this embodiment before being introduced into the cells may be, for example, 1 to 500 μg / mL or 10 to 100 μg / mL.

[0037] (Other Components) The pharmaceutical composition of this embodiment may contain, in addition to the cell population described above, a pharmaceutically acceptable carrier and / or additive. The blending ratio of the carrier or additive may be appropriately set based on the range commonly used in the pharmaceutical field.

[0038] The carrier is not particularly limited, and examples thereof include water, distilled water for injection, physiological saline, glucose solution, isotonic solution (e.g., solutions of sodium chloride, potassium chloride, glycerin, mannitol, sorbitol, boric acid, borax, propylene glycol, etc.), other aqueous solvents, and aqueous or oily bases, etc. These carriers may be used alone or in combination of two or more.

[0039] Examples of additives include, but are not limited to, stabilizers, solubilizers, suspending agents, emulsifiers, soothing agents, buffers, preservatives, antiseptics, pH adjusters, colorants, and absorption enhancers. Examples of stabilizers include, but are not limited to, albumin, globulin, gelatin, mannitol, glucose, dextran, ethylene glycol, propylene glycol, ascorbic acid, sodium bisulfite, sodium thiosulfate, sodium EDTA, sodium citrate, and dibutylhydroxytoluene. Examples of solubilizing agents include, but are not limited to, alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants (e.g., Polysorbate 80 (registered trademark), HCO-50, and the like). Examples of suspending agents include, but are not limited to, glycerin monostearate, aluminum monostearate, methylcellulose, carboxymethylcellulose, hydroxymethylcellulose, and sodium lauryl sulfate. Examples of emulsifiers include, but are not limited to, gum arabic, sodium alginate, and tragacanth. Examples of soothing agents include, but are not limited to, benzyl alcohol, chlorobutanol, and sorbitol. Examples of buffers include, but are not limited to, phosphate buffer, acetate buffer, borate buffer, carbonate buffer, citrate buffer, and Tris buffer. Examples of preservatives include, but are not limited to, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, chlorobutanol, benzyl alcohol, benzalkonium chloride, sodium dehydroacetate, sodium edetate, boric acid, and borax. Examples of antiseptics include, but are not limited to, benzalkonium chloride, parahydroxybenzoic acid, and chlorobutanol. Examples of pH adjusters include, but are not limited to, hydrochloric acid, sodium hydroxide, phosphoric acid, and acetic acid. These additives may be used alone or in combination of two or more.

[0040] (Administration) In this embodiment, a drug is encapsulated in particles containing a bioabsorbable polymer, the particles are introduced into cells that accumulate at the site of inflammation, and the drug is administered using the cells. This allows the drug to exert its effects at a lower dose than when the drug is administered alone. Therefore, the pharmaceutical composition of this embodiment is preferably used in a method that includes administering the drug at a dose lower than the dose per dosage unit of the drug. Here, the drug may be cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, irinotecan, or the like.

[0041] The pharmaceutical composition of this embodiment contains a predetermined amount of a drug per dosage unit. This predetermined amount is preferably smaller than the dosage per dosage unit when the drug is administered in a conventional manner, for example, when the drug is administered alone. Drugs targeted by the pharmaceutical composition of this embodiment include, for example, cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, and irinotecan, which are commercially available drugs. Furthermore, the dosage unit of each of these drugs is described in the package insert or the like in each country. A dosage of the pharmaceutical composition of this embodiment that is lower than the dosage per dosage unit of cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, irinotecan, or the like means that the dosage per dosage unit of cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, irinotecan, or the like when administered as the pharmaceutical composition of this embodiment is lower than the dosage per dosage unit of cyclosporine, minoxidil, docetaxel, paclitaxel, cisplatin, irinotecan, or the like when administered as a medical drug approved in each country. The predetermined amount may be, for example, 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, 8% or less, 5% or less, 3% or less, 2% or less, 1% or less, 0.8% or less, 0.5% or less, 0.3% or less, 0.2% or less, or 0.1% or less compared to the dose per dosage unit when the drug is administered in a conventional manner, for example, the drug is administered alone. The predetermined amount may be, for example, 0.1% or more, 0.2% or more, 0.3% or more, 0.5% or more, 1% or more, 2% or more, 3% or more, 5% or more, 8% or more, 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the dose per dosage unit when the drug is administered in a conventional manner, for example, the drug is administered alone. The predetermined amount may be, for example, within a range of 0.1 to 90% of the dose per dosage unit when the drug is administered in a normal manner, for example, when the drug is administered alone, or within a range in which the upper and / or lower limits of the range are arbitrarily replaced with the above-mentioned values.

[0042] The dosage and administration interval of the pharmaceutical composition of this embodiment can be appropriately selected depending on the type of drug, the subject to be administered, the administration route, the disease, and the age, weight, and symptoms of the subject. The pharmaceutical composition of this embodiment may be administered multiple times (e.g., 2 to 10 times), and the interval in this case is not particularly limited, and may be, for example, twice a day, once a day, twice a week, once a week, or once every two weeks. The pharmaceutical composition of this embodiment may also be administered periodically until the treatment of the disease is completed, and the interval in this case is also not particularly limited, and may be the interval described above. In particular, the administration interval of the pharmaceutical composition of this embodiment is not particularly limited, and may be, for example, once a week. The number of cells contained in one dosage unit of the pharmaceutical composition of this embodiment is not particularly limited, and may be, for example, 1 x 10 ３ ~1 x 10 １２ The number of cells contained in one dosage unit may be in the range of, for example, 1 x 10 4 The number of cells contained in one dosage unit may be 1 x 10 ３ pieces ~ 1×10 １２ Range of pieces, or 1 x 10 ４ pieces ~ 1×10 １２ In the range of 1 x 10 １１ It may be 1×10 or less, 10 The number of cells may be 1 or less. The above number of cells may be administered multiple times as a single dose, or this dose may be administered in multiple divided doses. The pharmaceutical composition of this embodiment may also be administered together with one or more other drugs. The content of the drug contained in the cells of the pharmaceutical composition of this embodiment is not particularly limited and is set depending on each drug, but may be, for example, 0.1 ng / 10 ５ The content may be 0.1 ng / 10 cells or more. ５ Cell ~1000ng / 10 ５ The lower limit of the content may be 0.3 ng / 10 ５ cells or more, and ５ cells or more, and ５ cells or more, and ５The content may be 500 ng / 10 cells or more within the range, or between the range and the lower limit. ５ It may be less than 100 ng / 10 ５ It may be less than 50 ng / 10 ５ Although not particularly limited, for example, when the drug is cyclosporine, the concentration may be 1 ng / 10 ５ The content may be 1 ng / 10 ５ Cell ~1000ng / 10 ５ The range may be 5 ng / 10 ５ Cell ~1000ng / 10 ５ The range may be 1 ng / 10 ５ cells ~100ng / 10 ５ The range may be 5 ng / 10 ６ cells ~100ng / 10 ５ The range may be 5 ng / 10 ６ Cell ~50ng / 10 ５ In addition, when the drug is minoxidil, the concentration may be in the range of 0.3 ng / 10 ５ The content may be 0.3 ng / 10 ５ Cell ~1000ng / 10 ５ The range may be 0.3 ng / 10 ５ cells ~100ng / 10 ５ The range may be 1 ng / 10 ５ Cell ~1000ng / 10 ５ The range may be 1 ng / 10 ５ cells ~100ng / 10 ５ The range may be 1 ng / 10 ５ Cell ~50ng / 10 ５ The administration interval, the dose, and the concentration of the drug described below may be appropriately selected and combined from among the modes described as possible in this specification.

[0043] The pharmaceutical composition and cells of this embodiment can be administered by direct application to tissue, but are not limited thereto. Examples of administration routes include intravascular administration, particularly intravenous administration, as well as intramuscular, intrathecal, intraperitoneal, intestinal, rectal, vaginal, intraocular, intracerebral, subcutaneous, nasal, sublingual, oral administration, inhalation, transdermal administration, implantation, spraying onto the surface of an organ, and direct (localized) administration to the affected area by application of a sheet or the like. Administration may be carried out, for example, as an injection or infusion, or by surgical implantation of a cell sheet or the like. Furthermore, when the cells of this embodiment are in the form of a sheet, the cell sheet of this embodiment may be fixed to the target tissue with a fastening means such as sutures or staples when applied to the tissue.

[0044] (Uses) The pharmaceutical composition of this embodiment can be used for various purposes depending on the type of drug encapsulated in the particles of this embodiment. For example, the pharmaceutical composition of this embodiment can be used for the treatment or prevention of a disease.

[0045] In one embodiment, when the drug is cyclosporine, i.e., when the particles of this embodiment contain cyclosporine, the pharmaceutical composition of this embodiment may be used to suppress immune responses. In this case, the pharmaceutical composition of this embodiment may be used to treat or prevent autoimmune diseases, inflammatory diseases, post-transplant rejection, etc. Specifically, the pharmaceutical composition of this embodiment may be used to suppress rejection in organ transplants such as kidney transplants, liver transplants, heart transplants, lung transplants, pancreas transplants, and small intestine transplants, suppress rejection and graft-versus-host disease in bone marrow transplants, suppress immune responses associated with cell transplants, and treat or prevent Behçet's disease and other non-infectious uveitis, psoriasis vulgaris, pustular psoriasis, psoriatic erythroderma, psoriatic arthritis, aplastic anemia, pure red cell aplasia, nephrotic syndrome, generalized myasthenia gravis, atopic dermatitis, systemic lupus erythematosus, multiple sclerosis, systemic sclerosis, autoimmune hepatitis, and dermatomyositis. Furthermore, the pharmaceutical composition of this embodiment may be used for the treatment or prevention of diseases known to be treatable or preventable with cyclosporine, or for diseases for which treatment or prevention with cyclosporine has been approved. In this case, the cells into which the particles of this embodiment are introduced preferably include peripheral blood mononuclear cells.

[0046] In the Examples described below, it was shown that immune cells (preferably peripheral blood mononuclear cells) transfected with cyclosporine-encapsulated PLGA particles exhibit a stronger immunosuppressive effect at lower concentrations than untreated immune cells. Therefore, the present specification provides a pharmaceutical composition for suppressing an immune response, comprising a population of immune cells and cyclosporine; the use of cyclosporine for enhancing the immunosuppressive effect of immune cells; and the use of cyclosporine in the manufacture of a pharmaceutical composition for suppressing an immune response. In this pharmaceutical composition, the concentration of cyclosporine may be 1 nM or more, 5 nM or more, 10 nM or more, 20 nM or more, 30 nM or more, 50 nM or more, 70 nM or more, 80 nM or more, or 90 nM or more. Examples of immune cells include those described above in the (Cells) section, and it is preferable to use mononuclear cells, particularly peripheral blood mononuclear cells.

[0047] In one embodiment, when the drug is minoxidil, i.e., when the particles of this embodiment contain minoxidil, the pharmaceutical composition of this embodiment may be used to promote angiogenesis. In this case, the pharmaceutical composition of this embodiment may be used for the treatment or prevention of diseases in which angiogenesis may be involved, alopecia, and wound healing. Specific examples of diseases in which angiogenesis may be involved include ischemic diseases, and examples of ischemic diseases include cardiovascular diseases or ischemic heart diseases (e.g., coronary artery disease, coronary artery thrombosis, myocardial infarction, angina pectoris, acute coronary syndrome, atrial fibrillation, sudden ischemic death, and transient ischemic attack), peripheral ischemic diseases or peripheral arterial diseases (e.g., peripheral occlusive arterial disease, lower limb arterial ischemic disease, distal limb ischemic disease, vascular embolism, venous thrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism, and wound healing in diabetic patients), and ischemic cerebral diseases (e.g., cerebral contusion, Parkinson's disease, multiple sclerosis, and cerebral infarction). Furthermore, the pharmaceutical composition of this embodiment may be used for the treatment or prevention of diseases known to be treatable or preventable with minoxidil, or for diseases for which minoxidil is known to be useful for treatment or prevention, or for diseases approved for treatment or prevention with minoxidil. In this case, the cells into which the particles of this embodiment are introduced preferably include mesenchymal stem cells. Examples of mesenchymal stem cells include the mesenchymal stem cells described above in the section (Cells), and it is particularly preferred to use adipose-derived mesenchymal stem cells.

[0048] In the Examples described below, it was shown that mesenchymal stem cells have a low angiogenesis-promoting effect when administered in small amounts to endothelial cells, but that angiogenesis is promoted when combined with minoxidil. Therefore, the present specification provides a pharmaceutical composition for promoting angiogenesis, comprising a population of mesenchymal stem cells and minoxidil; the use of minoxidil to enhance the angiogenesis-promoting effect of mesenchymal stem cells; and the use of minoxidil in the manufacture of a pharmaceutical composition for promoting angiogenesis. In this pharmaceutical composition, the concentration of minoxidil may be 1 nM or more, 5 nM or more, 10 nM or more, 20 nM or more, 30 nM or more, 50 nM or more, 70 nM or more, 80 nM or more, or 90 nM or more. Examples of mesenchymal stem cells include the mesenchymal stem cells described above in the (Cells) section, and adipose-derived mesenchymal stem cells are preferably used.

[0049] In one embodiment, when the drug is docetaxel, paclitaxel, cisplatin, or irinotecan, i.e., when the particles of this embodiment contain docetaxel, paclitaxel, cisplatin, or irinotecan, the pharmaceutical composition of this embodiment may be used to promote anti-tumor activity. In this case, the pharmaceutical composition of this embodiment may be used to treat or prevent diseases and cancers in which anti-tumor activity may be involved. Specifically, it may be used to treat or prevent breast cancer, prostate cancer, pancreatic cancer, gastric cancer, lung cancer, colon cancer, rectal cancer, colorectal cancer, small intestine cancer, esophageal cancer, brain tumor, schwannoma, liver cancer, kidney cancer, bile duct cancer, endometrial cancer, cervical cancer, uterine sarcoma, ovarian cancer, bladder cancer, urethral cancer, skin cancer, penile cancer, testicular tumor, and Kaposi's sarcoma. Furthermore, the pharmaceutical composition of this embodiment may be used for the treatment or prevention of diseases known to be treatable with docetaxel, paclitaxel, cisplatin, or irinotecan, or for diseases known to be treatable or preventable with docetaxel, paclitaxel, cisplatin, or irinotecan, or for the treatment or prevention of diseases approved for treatment or prevention with docetaxel, paclitaxel, cisplatin, or irinotecan. In this case, the cells into which the particles of this embodiment have been introduced preferably include mesenchymal stem cells.

[0050] The subject to which the pharmaceutical composition and the cell of this embodiment are administered may be a mammal. Mammals are not particularly limited, but include, for example, humans, non-human primates, domestic animals, laboratory animals, and livestock, with humans being particularly preferred.

[0051] [Cells] The cells of this embodiment are cells that accumulate at the site of inflammation, into which particles containing a bioabsorbable polymer and a drug have been introduced. The cells of this embodiment can be used as a pharmaceutical composition, optionally in the form of a cell population containing other cells. The cells of this embodiment can be used for the treatment or prevention of the above-mentioned diseases. The cells of this embodiment may be used in various forms, such as a cell suspension, a cell sheet, or a sheet-like cell culture. The cells of this embodiment are preferably used in a method that includes administering a drug at a dose smaller than the dose per dosage unit of the drug.

[0052] One aspect of this embodiment is the cell of this embodiment for use in a method comprising administering a drug at a dose less than the dose per dosage unit of the drug. One aspect of this embodiment is the use of the cell of this embodiment in a method comprising administering a drug at a dose less than the dose per dosage unit of the drug. One aspect of this embodiment is the cell of this embodiment for use in treating or preventing the disease. One aspect of this embodiment is the use of the cell of this embodiment in the method for treating or preventing the disease. One aspect of this embodiment is the use of the cell of this embodiment in the manufacture of the pharmaceutical composition of this embodiment.

[0053] [Particles] The particles of this embodiment are particles containing a bioabsorbable polymer and a drug, and are drug-encapsulated particles that are introduced into cells that accumulate at an inflammatory site. The particles of this embodiment are suitable for use in a method that includes administering a drug at a dose smaller than the dose per dosage unit of the drug.

[0054] An aspect of this embodiment is the particle of this embodiment for use in a method comprising administering a drug at a dose less than the dose per dosage unit of the drug. An aspect of this embodiment is the use of the particle of this embodiment in a method comprising administering a drug at a dose less than the dose per dosage unit of the drug. An aspect of this embodiment is the particle of this embodiment for use in treating or preventing the above-mentioned diseases. An aspect of this embodiment is the use of the particle of this embodiment in a method for treating or preventing the above-mentioned diseases. An aspect of this embodiment is the use of the particle of this embodiment in the manufacture of the pharmaceutical composition of this embodiment.

[0055] [Drug Administration Method] One aspect of this embodiment is a drug administration method that includes encapsulating a drug in a bioabsorbable polymer to prepare drug-encapsulated particles, introducing the drug-encapsulated particles into cells that accumulate at the site of inflammation, and administering the cells. This drug administration method of this embodiment allows for a reduction in the amount of drug administered per dosage unit compared to administering the drug by a conventional method, for example, administering the drug alone.

[0056] In the drug administration method of this embodiment, a drug is first encapsulated in a bioabsorbable polymer to prepare drug-encapsulated particles. Here, other components besides the drug may also be encapsulated in the drug-encapsulated particles. The bioabsorbable polymer, drug, other components, and method for encapsulating the drug in the bioabsorbable polymer are as described above. After the drug-encapsulated particles are prepared, the amount of drug encapsulated per drug-encapsulated particle may be measured.

[0057] Next, the drug-encapsulated particles are introduced into cells that accumulate at the site of inflammation. The cells that accumulate at the site of inflammation and the method of introduction are as described above. In the drug administration method of this embodiment, the cells that accumulate at the site of inflammation are preferably obtained from the subject to whom the drug is to be administered.

[0058] The cells into which the drug-encapsulated particles have been introduced are then administered to a subject. The administration subject, cell administration method, and dosage are as described above. The dosage per cell administration unit is preferably determined by measuring the amount and / or concentration of the drug contained in the administration product, and adjusting the dosage to a predetermined amount that is less than the dosage per administration unit when the drug is administered by a conventional method, for example, when the drug is administered alone. The predetermined amount is as described above. The amount and / or concentration of the drug contained in the administration product containing the cell population can be measured, for example, by a method combining liquid chromatography and mass spectrometry (e.g., LC / MS / MS).

[0059] The drug administration method of this embodiment may include selecting a drug depending on the disease of the subject to be administered the drug, before producing drug-encapsulated particles.

[0060] [Method of Treatment or Prevention] The pharmaceutical composition of this embodiment can be used for the treatment or prevention of the above-mentioned diseases. The pharmaceutical composition of this embodiment is preferably used for the treatment of the above-mentioned diseases. One aspect of this embodiment is a method of treating or preventing a disease, comprising administering a therapeutically effective amount of the pharmaceutical composition of this embodiment to a patient in need thereof. Another aspect of this embodiment is a method of treating or preventing a disease, comprising administering a therapeutically effective amount of the cells of this embodiment to a patient in need thereof. Another aspect of this embodiment is a method of treating or preventing a disease, comprising administering a drug to a patient in need thereof by the drug administration method of this embodiment. Here, the disease may be any of the above-mentioned diseases, and the drug contained in the pharmaceutical composition of this embodiment or the cells of this embodiment may be selected depending on the disease, and the drug to be administered by the drug administration method of this embodiment may be selected.

[0061] The present invention will be described in more detail below using examples and comparative examples, but the present invention is not limited to the following examples.

[0062] Example 1 (In vitro immunosuppression evaluation test using human peripheral blood mononuclear cells transfected with cyclosporine-encapsulated PLGA particles) Treatment of mononuclear cells with magnetic beads bearing CD3 / CD28 on their surface induces immune stimulation and promotes the proliferation of T cells contained in the mononuclear cells. The stimulated T cells were allowed to incorporate bromodeoxyuridine (BrdU) for a certain period of time, and T cell proliferation was evaluated by Cell ELISA using an anti-BrdU antibody.

[0063] A polymer solution was prepared by dissolving 50 mg of lactic acid-co-glycolic acid copolymer (PLGA) and 2.5 mg of cyclosporine in a mixture of 1 mL of acetone and 0.5 mL of ethanol. The polymer solution was added dropwise to 50 mL of 4 wt % polyvinyl alcohol (PVA) solution at room temperature while stirring at 400 rpm, yielding a suspension of PLGA particles encapsulating cyclosporine. After distilling off the acetone and ethanol, the mixture was centrifuged, and the precipitate was then collected and resuspended in distilled water. This centrifugation and resuspension procedure in distilled water was repeated three times. The final suspension was dispensed and stored at -80°C.

[0064] Next, a suspension of human peripheral blood mononuclear cells was prepared. A frozen tube of normal human peripheral blood mononuclear cells (Fujifilm Wako Pure Chemical Industries, Ltd. / 551-37651) was removed and thawed by immersing in a preheated incubator at 37°C. The thawed cell suspension was added to 10 mL of RPMI 1640 medium, mixed, and then centrifuged at 500 g for 10 minutes, and the supernatant was removed. The pelleted cells were washed again with RPMI 1640 medium and then suspended in RPMI 1640 medium to achieve an appropriate cell concentration.

[0065] The frozen cyclosporine-encapsulated PLGA particles (cyclosporine concentration 25 μg / mL) were thawed immediately before the test, and a mononuclear cell suspension (concentration 4×10 ５ cells / mL) at a volume ratio of 1:10. ２ The PLGA particles were taken up by the cells by treatment in the presence of PBS for 30 minutes, and then washed twice with PBS to remove PLGA particles that were not taken up by the cells.

[0066] Mononuclear cells transfected with cyclosporine-encapsulated PLGA particles and mononuclear cells not treated with PLGA particles (untreated mononuclear cells) were mixed at ratios of 0:100, 25:75, 50:50, and 100:0, respectively, to prepare four cell populations. Using a portion of the mononuclear cells that had incorporated cyclosporine-encapsulated PLGA nanoparticles, the cyclosporine concentration in the cells was measured by LC / MS / MS, and the amount of cyclosporine contained per cell was calculated. Based on the calculation results, the cyclosporine concentrations for each well were calculated to be 0 ng / mL, 10 ng / mL, 20 ng / mL, and 40 ng / mL, respectively. The cyclosporine content per cell was 0 ng (no cells), 1 ng / 1 x 10 ng, and 1 ng / 1 x 10 ng. ４ Cells, 2ng / 2x10 ４ Cells, 4ng / 4x10 ４ It was a cell.

[0067] These cell populations were seeded into each well of a 96-well plate, treated with CD3 / CD28 magnetic beads, and cultured for 24 hours, with the total number of cells in each well adjusted to be equal.

[0068] As controls, wells were prepared in which untreated mononuclear cells were added with 1000 ng / mL, 100 ng / mL, 10 ng / mL, or 0 ng / mL of cyclosporine, and wells in which untreated mononuclear cells were not treated with CD3 / CD28.

[0069] After 24 hours of culture, BrdU was added and the cells were cultured for an additional 6 hours. The cells were then fixed and lysed using the reagents in the BrdU Cell Proliferation Assay Kit (Merck / 2750). The DNA was then single-stranded and immobilized, and subjected to Cell ELISA using an anti-BrdU antibody. Two wells were tested for each treatment condition, and the average was used for analysis. The above test was performed three times independently, and the average and standard deviation were calculated to examine the results. The results are shown in Figure 1.

[0070] 1, it was found that in the control medium containing cyclosporine, the immunosuppressive effect of cyclosporine increased as the cyclosporine concentration increased to 10 ng / mL, 100 ng / mL, and 1000 ng / mL. Furthermore, it was found that peripheral blood mononuclear cells transfected with cyclosporine-encapsulated PLGA particles exhibited immunosuppressive effects even at lower cyclosporine concentrations compared to the control medium containing cyclosporine alone. In particular, a high immunosuppressive effect was observed even at a low cyclosporine concentration of 40 ng / mL.

[0071] Furthermore, under conditions containing 50% untreated mononuclear cells and 50% mononuclear cells transfected with cyclosporine-encapsulated PLGA particles, the immune activation value was predicted to be the average of the immune activation value obtained under conditions containing only untreated mononuclear cells and the immune activation value obtained under conditions containing only mononuclear cells transfected with cyclosporine-encapsulated PLGA particles, but the actual value was lower than the predicted value. The percentage difference between this predicted value and the actual value was calculated to be 40%. A similar calculation was performed under conditions containing 75% untreated mononuclear cells and 25% mononuclear cells transfected with cyclosporine-encapsulated PLGA particles, and the percentage difference between the theoretical value and the actual value was 30%.

[0072] Thus, when peripheral blood mononuclear cells transfected with cyclosporine-encapsulated PLGA particles were present in the presence of untreated mononuclear cells, the cyclosporine-encapsulated PLGA particle-transfected peripheral blood mononuclear cells exhibited an immunosuppressive effect greater than that of the peripheral blood mononuclear cells themselves, suggesting that the peripheral blood mononuclear cells transfected with cyclosporine-encapsulated PLGA particles also exert an immunosuppressive effect on surrounding untreated mononuclear cells via the release of cyclosporine.Furthermore, it was shown that a lower concentration of cyclosporine than that when cyclosporine is added alone exerts a sufficient effect.

[0073] Example 2 (In vitro angiogenesis evaluation test using mesenchymal stem cells transfected with minoxidil-encapsulated PLGA particles) When human umbilical vein endothelial cells (endothelial cells) are co-cultured with mesenchymal stem cells, the venous endothelial cells form a blood vessel-like structure. The formation of this blood vessel-like structure is achieved by co-culturing an excess amount of mesenchymal stem cells relative to the endothelial cells (e.g., three times the amount of endothelial cells). However, when the ratio of endothelial cells to mesenchymal stem cells is set to equal, the formation of the blood vessel-like structure is hardly observed. Below, the test was conducted under co-culture conditions where the ratio of endothelial cells to mesenchymal stem cells was set to equal.

[0074] By a method similar to that of Example 1, minoxidil-encapsulated PLGA particles were prepared by encapsulating minoxidil in PLGA.

[0075] Next, a suspension of mesenchymal stem cells was prepared. Human mesenchymal stem cells (ATCC / PCS-500-011 (normal human adipose-derived mesenchymal stem cells)) were expanded and cultured in a medium made by mixing equal volumes of KBM ADSC-2 (Kohjin Bio / 16030030) and 10% FBS-containing DMEM medium (the medium was changed every two days). When the cells had proliferated to an appropriate number, the medium was removed and the adherent cells were washed with PBS. TrypLE (Gibco / 12605010) was added and spread over the front of the adherent cells, after which the liquid was removed and the cells were incubated at 37°C, 5% CO ２ After confirming that the cells had detached, a cell suspension was obtained using an appropriate amount of medium.

[0076] A suspension of PLGA particles encapsulating minoxidil (PLGA particle concentration 10 mg / mL) and a suspension of mesenchymal stem cells (concentration 1 × 10 ５ cells / mL) were mixed at a 1:1 (volume ratio) and incubated at 37°C, 5% CO ２ The PLGA particles were taken up by the cells by treating them in the presence of HCl for 30 minutes. Four conditions were prepared, with the final PLGA particle concentrations being 5 mg / mL, 1.25 mg / mL, 0.5 mg / mL, and 0 mg / mL. The PLGA particles that were not taken up by the cells were then washed twice with PBS to remove them. The minoxidil content per cell was 6 ng / 1.5 x 10 ５Cells, 1.5ng / 1.5x10 ５ Cells, 0.6ng / 1.5x10 ５ Cells, 0ng / 1.5x10 ５ It was a cell.

[0077] Endothelial cells with fluorescently labeled cell membranes and mesenchymal stem cells were mixed in equal numbers and seeded into each well of a 96-well plate. The mixture was incubated at 37°C and 5% CO ２ After 48 hours of culture, the presence or absence of formation of blood vessel-like structures was observed and evaluated using a fluorescence microscope.

[0078] As a control, wells were prepared in which 100 μM, 25 μM, 10 μM, and 0 μM minoxidil was added under conditions in which equal amounts of mesenchymal stem cells not treated with PLGA particles (untreated mesenchymal stem cells) and endothelial cells were mixed.

[0079] The results of fluorescence microscopy under each condition are shown in Figure 2. No blood vessel-like structures were observed when vascular endothelial cells were cultured alone, or when vascular endothelial cells were cultured with 100 μM minoxidil. Furthermore, when vascular endothelial cells were cultured with three times the amount of mesenchymal stem cells, blood vessel-like structures were observed, but when the same amount of mesenchymal stem cells was co-cultured, almost no blood vessel-like structures were observed (Figure 2(A)).

[0080] Furthermore, when vascular endothelial cells and mesenchymal stem cells were co-cultured in equal amounts, the addition of 100 μM or 25 μM minoxidil promoted the formation of blood vessel-like structures in a concentration-dependent manner, while the addition of 10 μM minoxidil almost eliminated this effect (Figure 2(B)). Furthermore, under co-culture conditions in which an equal amount of mesenchymal stem cells into which minoxidil-encapsulated PLGA particles had been introduced was added instead of untreated mesenchymal stem cells, the formation of blood vessel-like structures was observed, and differences in the efficiency of blood vessel structure formation were observed depending on the concentration of minoxidil-encapsulated PLGA particles incorporated (Figure 2(B)).

[0081] The concentration of minoxidil in mesenchymal stem cells transfected with minoxidil-encapsulated PLGA particles was measured, and the concentration of minoxidil in the test system was calculated to be 50 nM when 5 mg / mL PLGA particles were incorporated. This indicates that the use of mesenchymal stem cells transfected with minoxidil-encapsulated PLGA particles provides sufficient effects at lower concentrations than when minoxidil is added alone.

Claims

1. A pharmaceutical composition comprising a population of cells, the cells being cells that accumulate at an inflammatory site into which particles containing a bioabsorbable polymer and a drug have been introduced, the drug being cyclosporine or minoxidil.

2. The pharmaceutical composition according to claim 1 for use in a method comprising administering cyclosporine or minoxidil in a dosage amount less than the dosage amount per dosage unit of cyclosporine or minoxidil.

3. The pharmaceutical composition according to claim 1 or 2, wherein the cells comprise at least one of immune cells and mesenchymal stem cells.

4. A pharmaceutical composition for suppressing an immune response according to claim 1 or 2, wherein the cells comprise peripheral blood mononuclear cells and the drug is cyclosporine.

5. The content of cyclosporine contained in the cells is 1 ng / 10 ５ The pharmaceutical composition of claim 4 which is a cell or more.

6. A pharmaceutical composition for suppressing an immune response, comprising a population of peripheral blood mononuclear cells and cyclosporine.

7. A pharmaceutical composition for promoting angiogenesis according to claim 1 or 2, wherein the cells comprise mesenchymal stem cells and the drug is minoxidil.

8. The content of minoxidil in the cells is 0.3 ng / 10 ５ The pharmaceutical composition of claim 7 which is a cell or more.

9. A pharmaceutical composition for promoting angiogenesis, comprising a population of mesenchymal stem cells and minoxidil.

10. A pharmaceutical composition for use in a method comprising: a population of cells, the cells being cells that accumulate at an inflammatory site into which particles comprising a bioabsorbable polymer and a drug have been introduced; and administering the drug at a dose less than the dose per dosage unit of the drug.

11. A particle for use in a method comprising administering a drug at a dose less than the dose per dosage unit of the drug, the particle comprising a bioabsorbable polymer and the drug, and being introduced into cells that accumulate at a site of inflammation.

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