WO2005089928A1 - Kit for extemporaneous preparation of coated fine particles - Google Patents

Abstract

A kit for the extemporaneous preparation of coated fine particles which comprises a fluid (fluid A) containing core fine particles dispersed therein and a polar organic solvent having a coating layer component constituting a coating layer for covering said core fine particles dissolved therein, and a solvent (fluid B) being able to mix with the fluid A and not containing the polar organic solvent or containing the polar organic solvent in a proportion than lower that in the fluid A, and a means for mixing the fluid A and the fluid B.

Description

 Specification

 Preparation kit for use of coated microparticles

 Technical field

 The present invention relates to a kit for preparing coated microparticles before use.

 Background art

 [0002] Conventionally, many techniques have been disclosed for a method for producing coated fine particles in pharmaceuticals, foods, agricultural chemicals, veterinary drugs, and the like. The coating of the fine particles (the fine particles to be coated) with the coating layer is, for example, to suppress the influence of the external factor force, selectively receive the effect of the external factor, and use the trigger as a trigger to change the fine particles. The purpose is to provide a function to fine particles. As one of the methods, a method of coating fine particles with a lipid membrane in a liquid has been reported (see Patent Document 1). In this method, the fine particles are coated with a lipid membrane by reducing the proportion of the polar organic solvent in the aqueous solution containing the polar organic solvent in which the fine particles are dispersed and the lipid is dissolved, and the coating is performed in the liquid. For example, coated microparticles suitable for intravenous injection microparticles and the like are produced with excellent efficiency.

 [0003] On the other hand, in the case of fine particle preparations produced by a method without coating, kits for preparation before use are known (for example, see Patent Documents 2 and 3).

 Patent Document 1: WO 02/28367 pamphlet

 Patent Document 2: JP-A-2000-247868

 Patent Document 3: Patent No. 2688235

 Disclosure of the invention

 Problems to be solved by the invention

[0004] It is an object of the present invention to provide a kit for preparation of coated microparticles in which core microparticles are coated with a coating layer.

 Means for solving the problem

[0005] The present invention relates to the following (1)-(16).

(1) The core fine particles are dispersed and the coating constituting the coating layer for coating the core fine particles is formed. Liquid containing a polar organic solvent in which the coating layer components are dissolved (liquid,

 A solvent that can be mixed with Liquid A and contains no polar organic solvent or contains a lower percentage of polar organic solvent than Liquid A (Liquid B),

 And a kit for preparing coated microparticles including a device provided with a means for mixing liquid A and liquid B.

 (2) Apparatus having means for mixing liquid A and liquid B means for storing liquid A, means for storing liquid B, in-line mixing means, and means for storing the liquid A (1) when using the coated fine particles according to (1) above, which is a device provided with a liquid sending means to the inlet of the line mixing means and a liquid sending means from the means for containing the liquid B to the inlet of the line mixing means. Preparation kit.

 (3) An instrument having a means for mixing the liquid A and the liquid B A syringe for storing the liquid A, a syringe for storing the liquid B, and a syringe for storing the liquid A and the liquid B The preparation kit for use of coated microparticles according to the above (1), which is a device provided with in-line mixing means having two inlets to which syringes can be connected respectively.

 (4) The device provided with a means for mixing the liquid A and the liquid B is a multi-chamber syringe having chambers for respectively storing the liquid A and the liquid B in parallel, and a means for discharging the locator in-line mixing of the syringe. A kit for preparing coated microparticles according to the above (1), which is a syringe.

(5) In-line mixing means The kit for preparing coated fine particles according to any one of (2) to (4), which is an in-line mixing means connected or built in with a static mixer.

(6) Instrument power with means for mixing liquid A and liquid B Means for containing liquid A, means for containing liquid B, and means for containing liquid B The kit for preparing a coated fine particle according to the above (1), wherein the kit comprises means for adding the liquid B to the liquid A in the liquid.

 (7) An instrument equipped with a means for mixing the liquid A and the liquid BA syringe for storing the liquid A and a syringe for storing the liquid B, and the two syringes are connected to each other to transfer the liquid B to the liquid A. The kit for preparing coated microparticles according to the above (1), which can be dried.

(8) The device provided with a means for mixing the liquid A and the liquid B is a multi-chamber syringe having chambers for accommodating the liquid A and the liquid B, respectively. Can be The kit for preparing coated microparticles according to (1) above, which is a possible syringe.

 (9) Device equipped with means for mixing liquid A and liquid B Force preparation at the time of use of coated fine particles according to any one of the above (2) to (8), which is a device equipped with a transfer needle or a connector for mixed injection at the outlet Kit

(10) The kit for use in preparing coated microparticles according to any one of (1) to (9) above, wherein the coating layer component is at least one substance selected from lipids, surfactants and polymers.

 (11) The kit according to any one of the above (1) to (10), wherein the coating layer is a lipid membrane.

 (12) The coated microparticle according to any one of (1) to (11), wherein the core microparticle is a microparticle having a drug, a lipid assembly, a ribosome, an emulsion microparticle, a polymer, a metal colloid, or a microparticle preparation as a constituent. Preparation kit before use.

 (13) The above-mentioned (1)-(11), wherein the core fine particle is a fine particle comprising a complex composed of a combination of two or more drugs, lipid aggregates, ribosomes, emulsion fine particles, polymers, metal colloids, or fine particle preparations. A kit for preparing coated microparticles according to any one of the above,

 (14) The kit for preparing coated microparticles according to any one of (1) to (11) above, wherein the core microparticles are microparticles comprising a complex of a drug and ribosome.

 (15) The kit for preparing the coated fine particles according to any one of (1) to (14) above, wherein the polar organic solvent is at least one selected from alcohols, glycols, and polyalkylene glycols.

 (16) Coated microparticles which can be prepared by the kit for preparing coated microparticles according to any one of (1) to (15) above.

 The invention's effect

 [0006] According to the present invention, a kit for preparing coated microparticles in which core microparticles are coated with a coating layer is provided.

 Brief Description of Drawings

FIG. 1 is a schematic view of one example of a kit of the present invention and its use (1).

 FIG. 2 is a schematic view of another example of the kit of the present invention and its usage (2).

FIG. 3 is a schematic view of another example of the kit of the present invention and its use (3). FIG. 4 is a schematic view of another example of the kit of the present invention and its use (4).

FIG. 5 is a schematic view of another example of the kit of the present invention and its usage (5).

FIG. 6 is a schematic view of another example of the kit of the present invention and its usage (6).

FIG. 7 is a schematic view of another example of the kit of the present invention and its usage (7).

FIG. 8 is a schematic view of another example of the kit of the present invention and its usage (8).

FIG. 9 is a schematic view of a conventional apparatus for producing coated fine particles.

 [FIG. 10] A schematic diagram of an example of the kit of the present invention and a method of using the same in the case where a kit for each use for preparing solution A and core fine particles at the time of use is included.

Explanation of symbols

1,2 containers

3,4 pump

5 Three-way joint

6 static mixer

7,8 infusion

9 Three-way joint

10 static mixer

11, 12 syringe

13 Three-way joint

14 static mixer

15 multi-chamber syringe

16 static mixer

17 containers

18 syringe

19 Transfer needle

20,21 infusion bottle

22 Mixed injection tube

Syringe with 23 female connectors

Syringe with 24 male connectors 25 series multi-chamber syringe

 26,27 containers

 28 pump

 29 containers

 30,31,32,33,34 syringe and transfer money 移

 35 syringe

 36 three-way fittings, static mixers and channels

 Liquid A A liquid containing a polar organic solvent in which core fine particles are dispersed and a coating layer component forming a coating layer for coating the core fine particles is dissolved.

 Liquid B Solvent that can be mixed with Liquid A and contains no polar organic solvent or contains a lower proportion of polar organic solvent than Liquid A

 liquid. Suspension obtained by mixing liquid A and liquid B

 BEST MODE FOR CARRYING OUT THE INVENTION

[0009] The kit for preparing coated fine particles of the present invention at the time of use comprises a liquid containing a polar organic solvent in which core fine particles are dispersed and a coating layer component constituting a coating layer for coating the core fine particles is dissolved. (Liquid A), a liquid that can be mixed with Liquid A, contains no polar organic solvent or contains a lower percentage of polar organic solvent than Liquid A (Liquid B), and has means for mixing Liquid A and Liquid B It is a kit that includes the equipment.

 [0010] The coated fine particles according to the present invention are fine particles formed by coating at least core fine particles and a coating layer, and a coating layer component constituting the coating layer covers the outside of the core fine particles. The fine particles are produced by mixing the liquid A and the liquid B with the above-mentioned device and coating the core fine particles with the coating layer component, and are obtained in the form of a suspension (liquid C).

[0011] The core fine particles in the present invention are, for example, fine particles comprising a drug, a lipid assembly, a ribosome, an emulsion particle, a polymer, a metal colloid, a fine particle preparation and the like, preferably at least a drug. And fine particles containing as a component. Further, the core fine particles according to the present invention may be a drug, a drug, a lipid assembly, a ribosome, an emulsion particle, a polymer, a metal colloid, or a complex obtained by combining two or more fine particle preparations as a component. Lipid aggregates, ribosomes, margillon particles, polymers, metal colloids, fine particles A complex obtained by combining a preparation or the like with another compound (e.g., a sugar, a lipid, an inorganic compound, or the like) may be used as a component, and fine particles containing a complex of a drug and a ribosome are preferable. .

 [0012] The drug includes a drug which takes the form of fine particles in the solvent of liquid A, a drug which forms a complex with other components constituting the core fine particles and takes the form of fine particles in the solvent of liquid A, and the like. Examples thereof include substances having pharmacological activity among proteins, peptides, nucleic acids, low molecular weight compounds, saccharides, high molecular weight compounds, lipid compounds, metal compounds, and the like, preferably nucleic acids, and more preferably nucleic acids. , Gene, DNA, RNA, oligonucleotide, plasmid and siRNA force.

 [0013] Examples of the protein or peptide include bradycune, angiotensin, oxytocin, vasopressin, adrenocorticotropin, calcitonin, insulin, glucagon, cholecystokun, β-endorphin, melanocyte inhibitor, melanocyte stimulating hormone, gastrin antagonist, neuron. Tensin, somatostatin, bunolecin, cyclosporine, enkephalin, transferrin, arginine-glycine-aspartic acid (RGD) peptide, thyroid hormone, growth hormone, gonadotropin, luteinizing hormone, asparaginase, anoreginase, pericase, canoleboxypeptidase, Speroxide dismutase, Itosori plasminogen activator, Streptokinase , Interleukin, interferon, muramyl dipeptide, thymopoietin, granulocyte colony stimulating factor, granulocyte macrophage colony stimulating factor, erythropoietin, trombopoetin, trypsin inhibitor, lysozyme, epidermal growth factor (EGF), insulin-like growth factor, nerve growth factor , Platelet-derived growth factor, transforming growth factor, endothelial cell growth factor, fibroblast (fibroblast) growth factor, glial cell growth factor, thymosin, specific antibody (for example, anti-EGF receptor antibody, etc.) And the like.

 [0014] Examples of the nucleic acid include oligonucleotides such as antisense oligonucleotides and sense oligonucleotides, genes, DNAs, RNAs, plasmids, siRNAs and the like. The nucleic acid is a phosphate moiety or an ester moiety in the nucleic acid structure. And the like, including derivatives substituted with another atom such as a sulfur atom. In addition, siRNA is short double-stranded

RNA. [0015] Examples of low molecular weight compounds include epsilon-aminocaproic acid, arginine hydrochloride, potassium L-asparaginate, tranexamic acid, bleomycin sulfate, vincristine sulfate, cefazolin sodium, cephalothin sodium, citicoline, cytarabine, gentamicin sulfate, Vancomycin hydrochloride, kanamycin sulfate, amikacin sulfate and the like.

 Examples of the saccharide include sodium chondroitin sulfate, sodium sodium phosphate, dextran fluorescein, and the like.

 Examples of the polymer compound include sodium polyethylene sulfonate, dibutyl ether-maleic anhydride copolymer (DIVEMA), styrene maleic anhydride copolymer-neocarzinostatin conjugate (SMANCS), and the like.

 Examples of the lipid conjugate include vitamin D, vitamin E and the like.

 Examples of the metal compound include cisbratin and the like.

 [0018] The lipid aggregate or ribosome is composed of, for example, a lipid and / or a surfactant. The lipid may be any of simple lipids, complex lipids, and derived lipids, such as phospholipids and glycemic lipids. Glycolipids, glycosphingolipids, sphingoids, sterols and the like are preferred, and phospholipids are preferred. Examples of the lipid include a surfactant (synonymous with the surfactant described below), a polymer (synonymous with the polymer described below, specifically, dextran or the like), a polyoxyethylene derivative (specifically, polyethylene). Glycol) and the like, and preferably a polyethylene glycolated phospholipid. Examples of the surfactant include a nonionic surfactant, an anionic surfactant, a cationic surfactant, an amphoteric surfactant and the like.

[0019] Examples of the phospholipid include phosphatidylcholine (specifically, soybean phosphatidylcholine, egg yolk phosphatidylcholine (EPC), distearoylphosphatidylcholine, dipalmitylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoylphosphatidylcholine, etc.), phosphatidyl Ethanolamine (specifically, distearoylphosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, etc.), glycerol phospholipids (specifically, phosphatidylserine, phosphatidic acid , Phosphatidylglycerol, phosphatidylinositol, lysophosphatidylcholine, etc.), sphingophospholipids (specifically sphingomyelin, sera Midophosphoethanolamine, ceramide phosphoglycerol, ceramide phosphoglycerol phosphoric acid, etc.), glycerol phosphonolipid, sphingo phosphonolipid, natural lecithin (specifically, egg yolk lecithin, soy lecithin, etc.), hydrogenated phospholipid (Specifically, natural or synthetic phospholipids such as hydrogenated phosphatidylcholine).

 [0020] Examples of the glycerol glycolipid include sulfoxyribosyl glyceride, diglycosyl diglyceride, digaratatosyl diglyceride, galactosyl diglyceride, glycosyl diglyceride and the like.

 Examples of glycosphingolipids include galactosyl celeb mouth side, ratatosyl cerebroside, and gandarioside.

 Examples of the sphingoides include sphingan, icosasus fingan, sphingocin, derivatives thereof, and the like when f is used. As the derivative, for example,-フ such as sphingan, icosasphingan, and sphingocin is represented by -NHCO (CH) CH (where x represents an integer of 0 to 18,

 2 2 3

 Among them, those converted to 6, 12, or 18 are preferable.

Examples of the sterols include, for example, cholesterol, dihydrocholesterol, lanosterol, β-sitosterol, campesterolone, stigmasterol, brassicasterol, ergocasterol, fucosterol, 3β- [Ν -(Ν'Ν'-Dimethylaminoethyl) potassium cholesterol (DC-Choi).

[0022] Other lipids include, for example, N- [l- (2,3-dioleoylpropyl)]-Ν, Ν, Ν-trimethylammonium chloride (DOTAP), N- [1 -(2,3-Dioleylpropyl)]-, Ν-dimethylamine (DODAP), N- [1- (2,3-Dioleyloxypropyl)]-Ν, Ν, Ν-trimethyl Ammonium chloride (DOTMA), 2,3-Dioleyloxy-N- [2- (sperminecarboxamido) ethyl] -Ν, Ν-dimethyl-1-propanami-dimethyltrifluoroacetic acid (DOSPA), N- [1- ( 2,3-ditetradecyloxypropyl)]-Ν, Ν-dimethyl-N-hydroxyethylammonium bromide (DMRIE), N- [l- (2,3-dioleyloxypropyl) ] Also included are lipids such as -Ν, Ν-dimethyl-N-hydroxyethylammonium bromide (DORIE).

Examples of the nonionic surfactant include polyoxyethylene sorbitan monooleate (specifically, polysorbate 80 and the like) and polyoxyethylene polyoxypropylene glycol (specifically, pull mouth nick F68 and the like) , Sorbitan fatty acids (specifically, sorbitan monolau Rate, sorbitan monooleate, etc.), polyoxyethylene derivatives (specifically, polyoxyethylene hydrogenated castor oil 60, polyoxyethylene lauryl alcohol, etc.), glycerin fatty acid esters and the like.

 [0024] Examples of the a-ionic surfactant include acyl sarcosine, sodium alkyl sulfate, alkyl benzene sulfonate, and sodium fatty acid having 7 to 22 carbon atoms. Specific examples include sodium dodecyl sulfate, sodium lauryl sulfate, sodium cholate, sodium deoxycholate, sodium taurodeoxycholate and the like.

 [0025] Examples of the cationic surfactant include an alkylamine salt, an acylamine salt, a quaternary ammonium salt, and an amine derivative. Specifically, benzalco-dimethyl salt, acylaminoethyl getylamine salt, N-alkyl polyalkyl polyamine salt, fatty acid polyethylene polyamide, cetyl trimethyl ammonium bromide, dodecyl trimethyl ammonium bromide, alkyl polyoxyethylene Amine, N-alkylaminopropylamine, fatty acid triethanolamine ester and the like.

 Examples of the amphoteric surfactant include 3-[(3-cholamidopropyl) dimethylammono] -1-propanesulfonic acid, N-tetradecyl-Ν, Ν-dimethyl-3-ammono-1 -Propanesulfonic acid and the like.

 [0027] In the ribosome, these lipids and surfactants are used alone or in combination, and preferably used in combination. When used in combination, the combination may be, for example, a combination of two or more components selected from the group consisting of hydrogenated soybean phosphatidylcholine, polyethylene glycol disulfide phospholipid and cholesterol, distearyl phosphatidyl choline, polyethylene glycolated phospholipid and cholesterol. Examples include a combination of two or more selected components, a combination of EPC and DOTAP, a combination of EPC, DOTAP and polyethylene glycolated phospholipid, a combination of EPC, DOTAP, cholesterol and polyethylene glycolated phospholipid, and the like.

 [0028] Further, the ribosome may contain a film stabilizer such as sterols such as cholesterol, for example, an antioxidant such as tocopherol if necessary.

[0029] Examples of the lipid aggregate include spherical micelles, spherical reverse micelles, sausage-like micelles, sausage-like reverse micelles, plate-like micelles, plate-like reverse micelles, hexagonal I, hexagonal II, and the like. And an aggregate comprising two or more molecules of lipid and lipid.

 Examples of the emulsion particles include oil-in-water (0 / W) emulsions and water-in-oil-in-water emulsions such as fat emulsions, emulsions comprising a nonionic surfactant and soybean oil, lipid emulsions, and lipid nanospheres. (W / 0 / W) emulsion particles.

 Examples of the polymer include natural polymers such as albumin, dextran, chitosan, dextran sulfate, and DNA, such as poly-L-lysine, polyethyleneimine, polyaspartic acid, styrene maleic acid copolymer, and isopropylacrylamide. Synthetic polymers such as acrylpyrrolidone copolymer, polyethylene glycol-modified dendrimer, polylactic acid, polylactic acid polydalicholate, polyethylene glycol iridani polylactic acid, and salts thereof.

 Here, the salt in the polymer includes, for example, a metal salt, an ammonium salt, an acid addition salt, an organic amine addition salt, an amino acid addition salt and the like. Examples of the metal salt include an alkali metal salt such as a lithium salt, a sodium salt, and a potassium salt, an alkaline earth metal salt such as a magnesium salt and a calcium salt, an aluminum salt, and a zinc salt. Examples thereof include salts such as ammonium and tetramethylammonium.Examples of the acid addition salts include inorganic salts such as hydrochloride, sulfate, nitrate and phosphate, and acetates. Organic acid salts such as maleate, fumarate, citrate and the like can be mentioned. Examples of the organic amine addition salts include addition salts such as morpholine and piperidine. And addition salts of phenylalanine, aspartic acid, glutamic acid, lysine, etc.

[0033] Examples of the metal colloid include metal colloids including gold, silver, platinum, copper, rhodium, silica, calcium, aluminum, iron, indium, cadmium, barium, lead, and the like.

 [0034] Examples of the fine particle preparation include microspheres, microcapsules, nanocrystals, lipid nanoparticles, polymer micelles, and the like.

When the core fine particle is a core fine particle containing a drug, preferably, the core fine particle contains a charged substance having a charge electrostatically opposite to that of the drug, and more preferably, a charged substance having an electrostatic charge opposite to that of the drug. It contains a lipid having an electrically opposite charge (cationic lipid or aionic lipid described below). Here, the charges electrostatically opposite to the drug include charges in the drug molecule, charges that generate an electrostatic attraction to intramolecular polarization, and the like, surface polarization, and the like. [0036] The charged substance contained in the core fine particles is classified into a cationic substance having a cationic property and an aionic substance having an ionic property. Even for amphoteric substances that have both groups, the relative negativity changes depending on the pH, binding with other substances, etc., so they are classified as cationic or ion-based depending on the occasion. Can be done. These charged substances may be used as constituents of the core fine particles or may be used after being added to constituents of the core fine particles.

 [0037] Examples of the cationic substance include those exemplified in the definition of the core fine particles described above. [Specifically, cationic lipids, cationic surfactants (as defined above), cationic polymers] Etc.], proteins or peptides capable of forming a complex at a pH below the isoelectric point.

 [0038] Examples of the cationic lipid include DOTAP, DODAP, DOTMA, DOSPA, DMRIE, DORIE, and DC-Choi.

 [0039] Examples of the cationic polymer include poly-L-lysine, polyethyleneimine, polyphenate (polyfect), chitosan and the like.

 [0040] The protein or peptide capable of forming a complex at a pH below the isoelectric point is a protein or peptide capable of forming a complex at a pH below the isoelectric point of the substance. Is not particularly limited. Examples include albumin, orosomucoid, globulin, fibrinogen, pepsin, ribonuclease T1 and the like.

 [0041] As the a-on substance, for example, an a-on substance among those exemplified in the definition of the core fine particles described above [specifically, an a-on lipid, an a-on surfactant (see above) Synonymous with), anionic polymer, etc.], proteins or peptides, nucleic acids, etc., capable of forming a complex at a pH above the isoelectric point.

 [0042] Examples of the a-on lipid include phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidic acid and the like.

Examples of the aionic polymer include polyaspartic acid, styrene-maleic acid copolymer, isopropylacrylamide-acrylpyrrolidone copolymer, polyethylene glycol-modified dendrimer, polylactic acid, polylactic acid polydaricholic acid, and polyethylene glycol i. Polylactic acid, dextran sulfate, dextran sodium sulfate, chondroitin sulfate, chondroitin Examples thereof include sodium sulfate, hyaluronic acid, chondroitin, deltaman sulfate, heparan sulfate, heparin, ketalan sulfate, dextranfluorescein acid and the like.

 [0044] A protein or peptide capable of forming a complex at a pH above the isoelectric point is a protein or peptide capable of forming a complex at a pH above the isoelectric point of the substance. Is not particularly limited. Examples include albumin, orosomucoid, globulin, fibrinogen, histone, protamine, ribonuclease, lysozyme and the like.

 [0045] Examples of the nucleic acid as the ion-containing substance include DNA, RNA, plasmid, siRNA, oligonucleotide, and the like. There may be.

The core fine particles can be produced as a commercially available product, or can be produced by a known method or a method similar thereto. For example, a known ribosome preparation method can be applied to the production of core microparticles containing ribosome, which is one of the core microparticles, as a component. Known methods for preparing ribosomes include, for example, the ribosome preparation method of Bangham et al. ["Journal of" Molecular Biology "(J. Mol. Biol.)", 1965, Vol. 13, p. 238-252], ethanol injection method [see "Journal of Cell Biol.", 1975, Vol. 66, p. 621-634], French press method [" FEBS Lett. ", 1979, Vol. 99, pp. 210-214], freeze-thaw method [" Arch. Biochem. Biophys. .) ", 1981, vol. 212, p. 186-194], reverse-phase evaporation [" Proceedings of the National Academy 'ob' Science 'United' States 'obb' U.S.A. (Proc. Natl. Acad. Sci. USA) ", 1978, Vol. 75, p. 4194-4198], pH gradient method (for example, Japanese Patent No. 2572554, Patent 2659136 Patent reference Publication), and the like. As a solution for suspending the ribosome in the production of the ribosome, for example, water, acid, alkali, various buffers, physiological saline, amino acid infusion and the like can be used. In the production of ribosomes, for example, antioxidants such as citric acid, ascorbic acid, cysteine, ethylenediaminetetraacetic acid (EDTA) and the like, for example, isotonic agents such as glycerin, glucose, sodium salt and the like are added. Is also possible. Alternatively, ribosomes can also be produced by dissolving fats or the like in an organic solvent such as ethanol, distilling off the solvent, adding physiological saline or the like, shaking, and forming ribosomes. it can.

[0047] Further, for example, a nonionic surfactant (as defined above), a cationic surfactant (as defined above), an ionic surfactant (as defined above), a polymer, a polyoxyethylene derivative and the like Ribosome surface modification can also be performed arbitrarily, and these surface-modified ribosomes are also used as a component of the core fine particles in the present invention [DD Lasic, Martin. Martin], edited by "Stealth 'Ribosomes". (Stealth Liposomes) "s United States," CRL Press Inc. (CRC Press Inc), 1995, pp. 93-102]. Examples of the polymer include dextran, pullulan, mannan, amylopectin, hydroxyethyl starch and the like. Examples of the polyoxyethylene derivative include polysorbate 80, pull-mouth nick F68, polyoxyethylene hydrogenated castor oil 60, polyoxyethylene lauryl alcohol, 1,2-distearoyl- _sn -glycerol-3-phosphoethanolamine-N -[Methoxy (polyethylene glycol) -2000] (PEG-DSPE).

 [0048] The average particle size of the ribosome can be freely selected as desired. Methods for adjusting the average particle size include, for example, an etastrusion method and a method of mechanically pulverizing large multi-membrane ribosomes (MLV) (specifically, using mantongoulin, microfluidizer, etc.) [Muller (RH Muller), Beta (S. Benita), Baume. Bohm), eds., "Emarzion 'and' Nano Suspensions' Fo ~~ · The 'Formulation' Ob Poir -Sorabu Nore Drags (See Emulsion ana Nanosuspensions for Tonne Formulation of Poorly Soluble Drugs, Scientific Publishers Stuttgart, Germany, 1998, pp. 267-294).

[0049] Further, a complex (for example, for example, a combination of two or more selected from drugs, lipid aggregates, ribosomes, emulsion particles, polymers, metal colloids, and fine particle preparations) constituting the core fine particles Complexes of ribosomes or lipid aggregates containing cationic lipids with nucleic acids, complexes of nucleic acids with polymers containing cationic polymers such as poly-L-lysine, and ionic properties of phosphatidic acid etc. Complex of lipid-containing ribosome or lipid aggregate and protein, complex of polymer containing anionic polymer such as styrene maleic acid and protein, ribosome or lipid aggregate containing cationic lipid and protein Complex with a protein, a polymer containing a cationic polymer such as poly-L-lysine, and a protein For example, a complex may be used.The production method may be, for example, a method in which a drug and a lipid assembly, ribosome, macromolecule, etc. are mixed in water. A conversion step and the like can be added. It is also possible to form a complex in various solvents such as acetone and ether. For example, a nucleic acid and a lipid can be dissolved in an organic solvent such as ethanol, the solvent can be distilled off, and then a physiological saline solution or the like can be added and shaken to form a nucleic acid complex. As another method for forming a complex, for example, a water-ionic substance and a polyethylene glycolated phospholipid (specifically, polyethylene glycol

-Phosphatidylethanolamine (more specifically, 1,2-distearoyl-sn-glycerol-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] (PEG-DSPE), etc.), poly Ribosomes are prepared using oxyethylene hydrogenated castor oil 60, CREMOPHORE (CREMOPHOR EL), etc., and then, for example, nucleic acid is added, and further, for example, a ionic polymer is added, and the multiplex complex is added. It is also possible.

 The size of the core fine particles is preferably several nm to several hundred μm, more preferably 10 nm to 5 μm, more preferably 50 nm to 300 nm. Most preferably, it is between 50 nm and 200 nm.

 [0051] Examples of the coating layer component include the lipids, surfactants, and polymers described in the definition of the core fine particles, and preferably the lipids and surfactants described in the definition of the core fine particles. One or more substances selected from surfactants may be mentioned, and more preferably, lipids and surfactants selected to form a lipid film for the coating layer, and one or more substances selected from surfactants, more preferably, Phospholipids.

[0052] Examples of the lipid used when the coating layer is a lipid membrane include, for example, synthetic lipids. Examples of the synthetic lipid include a fluorinated phosphatidylcholine, a fluorinated surfactant, a dialkylammonium bromide, and the like, and these may be used alone or in combination with other fats and the like. When the coating layer is a lipid membrane, it is preferable to use a water-soluble polymer derivative as one of the components of the coating layer. Examples of the water-soluble polymer derivative include, for example, polyethylene glycol dilipid [specifically, polyethylene dalicol-phosphatidylethanolamine (more specifically, 1,2-distearoyl-sn-glycero-3-phosphoethanola]. Min-N- [Methoxy (polyethylene glycol) -2000] (PEG-DSPE), polyoxyethylene hydrogenated castor oil 60, Cremophor

 (CREMOPHOR EL), polyethylene glycol sorbitan fatty acid esters (specifically, polyoxyethylene sorbitan monooleate, etc.), polyethylene glycol fatty acid esters, polyglycerin lipids (specifically, polyglycerin-phosphatidylethanolamine, etc.) ), Polyglycerin fatty acid esters and the like, and preferably polyethylene glycol iridani lipid.

[0053] The solvent in liquid A is preferably a solvent in which the core fine particles do not dissolve and the components of the coating layer dissolve. In liquid C, which is a mixture of liquid A and liquid B, the core fine particles do not dissolve, Layer components do not dissolve or aggregate. Liquid A is a liquid containing a polar organic solvent, Liquid B is a liquid containing a solvent other than the polar organic solvent, but Liquid A is a solvent containing a solvent other than the polar organic solvent except for the polar organic solvent in Liquid B. If the ratio is lower than the ratio, the liquid B may also contain the polar organic solvent if it is lower than the ratio of the polar organic solvent in the liquid A.

 In the present invention, “dispersing core fine particles” means that the core fine particles are suspended, emulsified or emulsified, preferably in a suspended state, and most of the core fine particles are dispersed, The state includes a state in which the remaining part is dissolved or a state in which a part is precipitated, but it is preferable that almost all or all of the core fine particles are dispersed. In addition, the term “dissolution of the coating layer component” includes a state in which most of the coating layer component is dissolved and the remaining portion is dispersed, but it is necessary that almost all or all of the coating layer component is dissolved. I like it.

[0055] Examples of the polar organic solvent in the liquid A and the liquid B include alcohols such as methanol, ethanol, n-propanol, 2-propanol, n-butanol, 2-butanol and tert-butanol, glycerin, ethylene glycol, and the like. Glycols such as propylene glycol and the like, polyalkylene glycols such as polyethylene glycol and the like are preferable, and ethanol is preferable. Examples of the solvent other than the polar organic solvent in the liquid A and the liquid B include water, liquid carbon dioxide, liquid hydrocarbon, halogenated carbon, halogenated hydrocarbon, and the like, and preferably water. Can be Further, the liquid A and the liquid B may contain ions, buffer components and the like. [0056] The combination of the polar organic solvent and the solvent other than the polar organic solvent is preferably a combination that can be mixed with each other. The selection can be made in consideration of the solubility of the components of the coating layer. On the other hand, it is preferable that the core fine particles have low solubility in any of the solvents in the liquids AC, and the polar organic solvent and the solvent other than the polar organic solvent also have low solubility in deviation. The coating layer component, which is preferred, preferably has low solubility in the solvent in liquids B and C, and preferably has high solubility in the solvent in liquid A. It is preferable that the solubility in a solvent other than the polar organic solvent, in which the solubility in a polar organic solvent is preferably high, is low.

 [0057] The proportion of the polar organic solvent in the solvent in the liquid A is such that the core fine particles are present without being dissolved and the coating layer component covering the core fine particles is dissolved! The force varies depending on the type of the solvent, the core fine particles, and the components of the coating layer, but is not particularly limited. The force is preferably 30 vol% or more, and more preferably 60 to 90 vol%. The ratio of the polar organic solvent in the solvent in the liquid C is not particularly limited as long as the core fine particles can be covered with the covering layer component in which the core fine particles are dissolved, but is preferably 50 vol% or less. , More preferably 30 to 50 vol%.

 [0058] The combination of the core fine particle and the coating layer component in the present invention is not particularly limited, but the core fine particle is a fine particle comprising a complex of a drug and a ribosome, and the coating layer component is lipid and / or surfactant. The combination that is the agent is preferred. The core microparticles are microparticles comprising a liposome or a complex of a drug and a ribosome as a component, the coating layer component is a lipid and / or a surfactant, and the coating layer is a lipid membrane. The force is also classified as ribosome in a narrow sense, and the core fine particles are other than fine particles composed of ribosome or a complex of drug and ribosome, and the coating layer component is lipid and / or surfactant, and the coating layer is lipid membrane. Is classified as a ribosome in a broad sense. In the present invention, the coated fine particles are more preferably ribosomes in a narrow sense.

[0059] The ratio of the core fine particles to liquid A and liquid C used in the kit for preparing coated fine particles of the present invention at the time of use is particularly limited as long as the core fine particles can be coated with the coating layer component. Although not particularly limited, 1 μg / mL-lg / mL is preferred, and 0.1-500 mg / mL is more preferred. Further, the ratio of the coating layer components (eg, lipids and the like) used to liquid A and liquid C is not particularly limited as long as the core fine particles can be coated, but is preferably 1 μg / mL-lg / mL, and 0.1%. — 400 mg / mL is more preferred. The ratio of the coating layer component to the core fine particles is preferably from 1: 0.1 to 1: 1000 by weight, more preferably from 1: 1 to 1:10.

 The liquid A in the kit for preparing coated fine particles of the present invention at the time of use may be prepared at the time of use. As a method for preparing the liquid A at the time of use, for example, a liquid (liquid D) in which core fine particles are dispersed is prepared, and a liquid (liquid E) in which the coating layer component is dissolved in a solvent containing a polar organic solvent is prepared. And a method of mixing the liquid D and the liquid E.More preferably, a liquid in which the core fine particles are dispersed (liquid D), a liquid in which the coating layer component is dissolved in a solvent containing a polar organic solvent (liquid E), And a method of preparing using a ready-to-use preparation kit having a means for mixing solution D and solution E. Here, each solvent in solution D and solution E is used as solution A after mixing. Such a solvent may be used as long as the core fine particles are dispersed and the coating layer components are dissolved.

 [0061] Further, the core fine particles in the liquid D are also prepared at the time of use, and examples of the method for preparing the core fine particles include the method for producing the core fine particles described in the description of the core fine particles. More preferably, there is a method for producing a complex in which two or more selected from drugs, lipid aggregates, ribosomes, emulsion particles, polymers, metal colloids, fine particle preparations and the like constituting the core fine particles are combined. It is preferable to prepare each using a preparation kit for each use.

 The kit for preparing the coated fine particles of the present invention at the time of use may include the kit for preparing the liquid A at the time of use and, if desired, the kit at the time of preparing the core fine particles.

[0063] Examples of the device of the present invention provided with a means for mixing the liquid A and the liquid B include a means for containing the liquid A, a means for containing the liquid B, an in-line mixing means, and a means for mixing the liquid A. A device having a means for feeding the liquid A to the inlet of the in-line mixing means and a means for feeding the liquid B to the inlet of the in-line mixing means. Means for containing liquid B, means for containing liquid B, and means for containing liquid B. Equipment including means for adding liquid B to liquid A in means for containing liquid A. Can be The device may include a mixing means that may include a flow path. Ma The outlet of the device may be provided with a transfusion needle or a co-infusion connector.

 Examples of the in-line mixing means include a T-tube, a Y-tube, a three-way joint, a three-way cock, an in-line mixer, an in-line mixer assembly, and the like. Like,.

 [0065] Examples of the means for storing the liquid A and the means for storing the liquid B include vials, infusion bags, and syringes, each of which may include a mixing means.

 [0066] Examples of the liquid sending means include a gear pump, a tube pump, a syringe pump, a suction pump, a pressurizing pump, a plunger type pump, and the like, and include a manually pressed syringe and a natural fall utilizing a height difference. In this case, the liquid sending means may also function as a flow path. The flow path may be such a flow path as a metal pipe or a resin tube, for example.

 Hereinafter, a kit for preparing coated fine particles of the present invention at the time of use and a method for producing coated fine particles using the kit will be described with reference to the drawings. However, the kit for preparing coated fine particles of the present invention at the time of use is not limited to those shown in these figures.

 FIG. 1 is a schematic view of the kit of the present invention, which is an instrument having a means for mixing liquid A and liquid B, means for storing liquid A, means for containing liquid B, in-line mixing means, and liquid A Is a device provided with a liquid sending means to the inlet of the in-line mixing means and a means for feeding the liquid B to the inlet of the in-line mixing means. Examples and their usage are given. The kit is

 I. Liquid A,

 液. Liquid B, and

 III. Container containing liquid A (1), container containing liquid B (2), pumps (3) and (4), three-way joint (5), static mixer (6), and channel force.

 To prepare coated microparticles using the kit, solution A in (1) is sent to (5) by (3), and solution B in (2) is converted to (5) by (4). Send liquid. The liquid A and the liquid B are mixed in (5) and (6), and the coated fine particles are obtained as a suspension (liquid C).

[0068] FIG. 2 is a schematic view of the kit of the present invention, which is an instrument having a means for mixing liquid A and liquid B. A means for storing liquid A, a means for storing liquid B, an in-line mixing means, A means for feeding the liquid A from the means for containing the liquid A to the inlet of the in-line mixing means and the liquid B; Means for accommodating the container Another example of a kit which is a device provided with a liquid feeding means to an inlet of the in-line mixing means, and a usage thereof are shown. The kit is

 I. Liquid A,

 液. Liquid B, and

 III. Infusion bottle containing liquid A (7), infusion bottle containing liquid B (8), three-way joint (9), static mixer (10), and flow path force.

 To prepare coated microparticles using the kit, solution A in (7) is sent to (9) by height difference, and solution B in (8) is sent to (9) by height difference I do. The liquid A and the liquid B are mixed in (9) and (10), and the coated fine particles are obtained as a suspension (liquid C).

[0069] FIG. 3 shows a kit of the present invention, which is also an instrument having a means for mixing liquid A and liquid B. A means for storing liquid A, a means for storing liquid B, an in-line mixing means, A kit which is a device provided with a liquid sending means from the means for containing the liquid A to the inlet of the in-line mixing means and a means for holding the liquid B to the inlet of the in-line mixing means. Other examples and uses thereof are shown, in which the device comprises, specifically, a syringe for containing liquid A, a syringe for containing liquid B, and a syringe for containing liquid A. This is an instrument equipped with in-line mixing means having two inlets to which syringes for storing liquid B can be connected. The kit is

 I. Liquid A,

 液. Liquid B, and

 III. A syringe containing liquid A (11), a syringe containing liquid B (12), a three-way joint (13), a static mixer (14), and a flow path are also provided.

 To prepare coated microparticles using the kit, liquid A in (11) is sent to (13) by pressing the plunger, and liquid B in (12) is pressed by pressing the plunger ( Send to 13). The liquid A and the liquid B are mixed in (13) and (14), and the coated fine particles are obtained as a suspension (liquid C).

[0070] FIG. 4 shows the kit of the present invention, which is also an instrument having means for mixing liquid A and liquid B. means for storing liquid A, means for storing liquid B, means for in-line mixing, A means for feeding the liquid A from the means for containing the liquid A to the inlet of the in-line mixing means and the liquid B; Means for containment Another example of a kit which is a device provided with a means for feeding liquid to the inlet of the in-line mixing means and its use are shown, and the devices are specifically liquid A and liquid This is a multi-chamber syringe having chambers for accommodating B in parallel, and a syringe serving as a discharge-loca in-line mixing means for the syringe. The kit is

 I. Liquid A,

 液. Liquid B, and

 III. Multi-chamber syringe (15) and static mixer (16)

 To prepare coated microparticles using the kit, liquid A in one chamber of (15) and liquid B in another chamber of (15) are sent to (16) by pressing a plunger. Liquid. Liquid A and liquid at (16)

B is mixed to obtain coated fine particles as a suspension (liquid C).

[0071] FIG. 5 is a diagram showing a device of the kit of the present invention, which includes a means for mixing liquid A and liquid B, means for storing liquid A, means for containing liquid B, and liquid B. A kit that is provided with a means for mixing liquid B into liquid A in the means for storing liquid A from the means for storing

An example and its use are shown. The kit is

 I. Liquid A,

 液. Liquid B, and

 III. Vessel containing liquid A (17), syringe containing liquid B (18) and infusion needle (19) Is sent to (17) through (19) by pressing the plunger. Liquid B sent to liquid A in (17) is added, and the coated fine particles are obtained as a suspension.

[0072] FIG. 6 shows a kit of the present invention, which also includes a device for mixing liquid A and liquid B, a device for storing liquid A, a device for storing liquid B, and a device for storing liquid B. Means for accommodating a container Another example of a kit which is a device provided with a unit for kneading a liquid B into a liquid A in a unit for accommodating the liquid A, and the usage thereof are shown. The kit is

 I. Liquid A,

 液. Liquid B, and

III. Infusion bottle containing solution A (20), infusion bottle containing solution B (21) and mixed injection tube (22).

 To prepare coated microparticles using the kit, solution B in (21) is sent to (20) via (22) at a height difference. Liquid B sent to liquid A in (20) is added, and the coated fine particles are obtained as a suspension.

 [0073] FIG. 7 shows a kit of the present invention, which is an instrument having a means for mixing liquid A and liquid B. A means for containing liquid A, a means for containing liquid B, and a liquid B This shows other examples of the kit which is a device provided with a means for converting the solution B from the means for containing the solution A to the solution A in the means for containing the solution A, and the usage thereof. Specifically, a syringe for containing the liquid A and a syringe for containing the liquid B. In the two syringes, it is possible to add the liquid B to the liquid A by connecting the two syringes to each other. . The kit is

 I. Liquid A,

 液. Liquid B, and

 III. Syringe (23) containing liquid A with female connector at outlet and syringe (24) containing liquid B with male connector at outlet.

 To prepare coated microparticles using the kit, connect the female connector of (23) and the male connector of (24), and send the liquid B in (24) to (23) by pressing the plunger. I do. The liquid B sent to the liquid A in (23) is added, and the coated fine particles are obtained as a suspension.

[0074] FIG. 8 is a schematic view of the kit of the present invention, which is an instrument having a means for mixing liquid A and liquid B. means for storing liquid A, means for containing liquid B, and liquid B. This shows other examples of the kit which is a device provided with a means for converting the solution B from the means for containing the solution A to the solution A in the means for containing the solution A, and the usage thereof. More specifically, this is a multi-chamber syringe that has chambers for accommodating liquid A and liquid B, respectively, and is capable of pumping liquid B to liquid A. The kit is

 I. Liquid A,

 液. Liquid B, and

 III. In-line multi-chamber syringe (25) Power is also increased.

To prepare coated microparticles using the kit, press the plunger (25) to move the indoor gasket (a in the figure) to the bypass (b in the figure), and then press the plunger Thus, the liquid B in the plunger-side chamber is sent to the outlet-side chamber through the bypass. The sent liquid B is added to the liquid A in the outlet side chamber, and the coated fine particles are obtained as a suspension.

 [0075] On the other hand, Fig. 9 shows a coated fine particle producing apparatus according to a conventional technique (see Patent Document 1), in which a container (26) containing liquid A, a container (27) containing liquid B, and a pump ( 28) represents an apparatus for producing coated fine particles provided with 28). In order to prepare coated fine particles using the production apparatus, the liquid B in (27) is sent to (26) by (28). The liquid A in (26) and the liquid B sent are mixed to obtain coated fine particles as a suspension.

 [0076] The syringe used in the present invention may be specially prepared, but a commercially available syringe can also be used and is not particularly limited. Specifically, one-component syringes (sold by Terumo, Nipro, etc.), two-component syringes (sold by Vetter (Germany), Betaton's Dickinson (US), etc.), two-component mixed types Syringes (commercially available from TAH Industries (USA), Plas-Pak Industries (USA), etc.) and the like, and liquid A and / or liquid B may be pre-filled as a prefilled syringe. The mixing ratio of liquid A and liquid B can be arbitrarily changed by combining syringes having different inner diameters.

 [0077] Next, the preparation method of coated microparticles in the case where the kit for preparing coated microparticles of the present invention at the time of use includes a kit for preparing solution A and core microparticles at the time of use is described below. In the case where the microparticles are a complex of a drug and ribosome and the coating layer component is a lipid, a kit for preparing coated microparticles at the time of use will be specifically described as an example. However, the time preparation kit for preparing the solution A and the core fine particles at the time of use in the time of use preparation kit of the coated fine particles of the present invention is not limited thereto.

 • Composition of fresh preparation kit (Figure 10)

 Raw material for core fine particles; a. Ribosome or raw material for ribosome

 b. Drug (preferably, aqueous solution of drug)

 Solvent for liquid D; a. Water

 b. Polar organic solvent

Liquid E; a liquid containing a polar organic solvent in which lipid is dissolved (preferably, an aqueous solution containing the same or different polar organic solvent as solvent b of liquid D, more preferably an aqueous solution containing the same polar organic solvent ) Liquid B; water

 Equipment; a. A container for containing ribosomes or ribosome raw materials, and for preparing liquid A at the time of use (eg, vial) (29)

 b. Syringe containing drug (preferably, drug aqueous solution) and needle for liquid transfer (30) c Syringe containing solvent a for solution D of liquid D and needle for liquid transfer (31)

 d. Syringe containing solvent b of liquid D and needle for liquid transfer (32)

 e. Syringe containing liquid E and needle for liquid transfer (33)

 f. Syringe for containing liquid A and needle for liquid transfer (34)

 g.Syringe containing liquid B (35)

 h. Three-way fittings, static mixers and channels (36)

Method for Preparing Coated Fine Particles

 Add the solvent a of solution D in (31) to the ribosome or raw material of ribosome in (29) and shake, then add the drug in (30) to (29) and shake to mix. Then, core fine particles are prepared. Similarly, the solution A is prepared by putting the solvent b of the solution D in (32) and the solution F in (33) in this order in (29) and shaking to mix. The obtained liquid A is transferred to (34) and connected to (36) as shown in FIG. Similarly, the syringe (35) containing liquid B is connected to (36) as shown in Fig. 10, and the plungers of the syringes are pressed to send liquid A and liquid B to (36), respectively. Liquid A and liquid B are mixed to obtain coated fine particles as a suspension (liquid C).

[0079] The size of the coated fine particles of the present invention is preferably an average particle size of 100 mm or less, more preferably 200 mm or less, and more specifically, for example, an injectable size. Is preferred.

 [0080] The coated fine particles of the present invention can be used for stabilizing a drug against biological components such as blood components, gastrointestinal fluid, etc., reducing side effects, increasing drug accumulation on a target organ such as a tumor, orally or transmucosally. Can be used as a preparation for the purpose of improving the absorption of a drug at the same time. For the purpose, the coated fine particles of the present invention are preferably coated fine particles containing a drug firmly and capable of encapsulating the drug in a biological component for a long time.

When the coated fine particles of the present invention are used as a preparation, the coated fine particle suspension prepared by the above method can be used as it is, for example, in the form of an injection or the like. The suspension power can be used after removing the solvent by, for example, filtration or centrifugation.

 [0082] In the case of an injection, the suspension of the coated fine particles of the present invention or the coated fine particles from which the solvent has been removed is combined with, for example, water, acid, alkali, various buffers, physiological saline, and amino acid infusion. And the like are preferably mixed to prepare an injection. Also, an injection can be prepared by adding an antioxidant such as citric acid, ascorbic acid, cysteine, and EDTA, and an isotonic agent such as glycerin, glucose, sodium salt and the like. In addition, for example, a cryopreservative such as glycerin can be added and stored frozen.

 [0083] In addition, the suspension or the injection may be used for infusion or infusion using, for example, a physiological saline bottle, a double bag kit (Nipro), a drug vial storage type full kit (Nipro), or the like. It is also possible to mix with various solutions. In this case, it is preferable that the device provided with a means for mixing the liquid A and the liquid B is a device provided with a transfer needle or a co-injection connector at the outlet.

Next, the present invention will be specifically described with reference to examples. However, the present invention is not limited to these examples.

 Example 1

 Using the following kit, solution A and solution B, liquid A was mixed at 5.2 mL / h and solution B at ll. LmL / h. After mixing, the ethanol concentration in solution C was about 20 vol%. The liquid was sent. At the outlet of liquid C of the assembled kit, a reservoir containing 18.8 mL of distilled water was placed in advance, and the obtained suspension of the coated fine particles was dropped. After sending the solution for 23 minutes, the reservoir was covered and inverted mixed, and the resulting mixture was subjected to ultracentrifugation (1 hour, 110,000 X g, 25 ° C). The precipitate after ultracentrifugation was resuspended by adding phosphate buffered saline (PBS) so that the total lipid concentration after dilution was 30 mg / mL, to obtain a preparation.

 Yat;

The kit shown in FIG. 3 was used. Use a three-way fitting for HPLC piping (manufactured by Shimadzu Corporation) for (13), and use a 2 cm static mixer [14 mm) in a fluorine resin tube with a length of 20 mm and an inner diameter of 1.5 mm. )), And a 20 mL syringe (manufactured by Terumo) loaded in a syringe pump (manufactured by Nikkiso) was used in (11) and (12). Liquid A; Dextran Fluororesinonic (FD) (Molecular Probes; manufactured by Molecular Probes; the same applies hereinafter) 30 mg, DOTAP (Avanti Polar Lipids); the same applies hereinafter 9) Distilled water (9 mL) was added to 180 mg and PEG-DSPE (72 mg), and the mixture was shaken with a vortex mixer. The obtained suspension was passed through a 0.4 m polycarbonate membrane filter 10 times and a 0.1 μm polycarbonate membrane filter 10 times at room temperature to obtain a suspension of core fine particles. 750 L of the obtained suspension of core fine particles and 1 mL of ethanol were mixed, and 480 mg of EPC and 100 mg of PEG-DSPE were mixed with 250 mL of ethanol, and 250 μL of a solution obtained by stirring was mixed.

 Liquid Β;

 Distilled water

 Example 2

 [0086] Using the same kit, liquid Α and liquid と as in Example 1, the concentration of ethanol in liquid C after mixing liquid A at 5.2 mL / h and liquid B at 5.6 mL / h was approximately 30 vol%. Respectively. At the outlet of liquid C of the assembled kit, a reservoir containing 20.9 mL of distilled water in advance was placed so that the suspension of the obtained coated fine particles was dropped. After feeding the solution for 23 minutes, mixing, ultracentrifugation, and resuspension were performed in the same manner as in Example 1 to obtain a preparation.

 Example 3

 [0087] Using the same kit as in Example 1, solution A and solution B, the concentration of ethanol in solution C after mixing solution A at 5.2 mL / h and solution B at 2.9 mL / h was approximately 40 vol%. Respectively. At the outlet of liquid C of the assembled kit, a reservoir containing 21.9 mL of distilled water in advance was placed so that the obtained suspension of coated fine particles was dropped. After feeding the solution for 23 minutes, mixing, ultracentrifugation, and resuspension were performed in the same manner as in Example 1 to obtain a preparation.

 Example 4

Using the same kit as in Example 1, solution A and solution B, the concentration of ethanol in solution C after mixing solution A at 5.2 mL / h and solution B at 0.7 mL / h was approximately 55 vol%. Respectively. At the outlet of liquid C of the assembled kit, a reservoir containing 22.7 mL of distilled water in advance was placed so that the obtained suspension of coated fine particles was dropped. After feeding the solution for 23 minutes, mixing, ultracentrifugation, and resuspension were performed in the same manner as in Example 1 to obtain a preparation. Example 5

 Using the same kit as in Example 1, solution A and solution B, the ethanol concentration in solution C after mixing solution A at 26.0 mL / h and solution B at 28.0 mL / h is approximately 30 vol%. Respectively. At the outlet of liquid C of the assembled kit, a reservoir containing 20.9 mL of distilled water in advance was placed so that the obtained suspension of coated fine particles was dropped. After feeding for 4.6 minutes, the mixture was mixed, ultracentrifuged and resuspended in the same manner as in Example 1 to obtain a preparation.

 Example 6

 [0090] Using the same kit as in Example 1, solution A and solution B, the concentration of ethanol in solution C after mixing solution A at 1.0 mL / h and solution B at 1.1 mL / h is approximately 30 vol%. Respectively. At the outlet of liquid C of the assembled kit, a reservoir containing 20.9 mL of distilled water in advance was placed so that the suspension of the obtained coated fine particles was dropped. After feeding for 115 minutes, the mixture was mixed, ultracentrifuged and resuspended in the same manner as in Example 1 to obtain a preparation.

 Example 7

 [0091] Using the same kit as in Example 1 except for the static mixer and the same liquid A and liquid B as in Example 1, liquid A was mixed at 5.2 mL / h, and liquid B was mixed at 5.6 mL / h. Each solution was fed such that the ethanol concentration in Solution C became approximately 30 vol%. A reservoir containing 20.9 mL of distilled water in advance was placed at the outlet of the liquid C of the kit which had been set up so that the suspension of the obtained coated fine particles was dropped. After feeding the solution for 23 minutes, the mixture was mixed, ultracentrifuged and resuspended in the same manner as in Example 1 to obtain a preparation.

 Example 8

 [0092] Using the following kit, solution A and solution B, each solution was fed at 5.2 mL / h. At the outlet of liquid C of the assembled kit, a reservoir containing 21.0 mL of distilled water in advance was placed so that the obtained suspension of the coated fine particles was dropped. After feeding for 23 minutes, the mixture was mixed, ultracentrifuged and resuspended in the same manner as in Example 1 to obtain a preparation.

 Yat;

The kit shown in FIG. 4 was used. In (15), a two-liquid mixed syringe (TAHplus 9mL Micro dispenser) loaded into a syringe pump (manufactured by Nikkiso Co., Ltd.) (Ispencer 1): US-T.H. Co., Ltd.) and (16) a static mixer (spiral mixer 190-212: US-T.H. Co., Ltd.) were used.

Liquid A;

 9 mL of distilled water was added to 30 mg of FD, 180 mg of DOTAP and 72 mg of PEG-DSPE, and the mixture was shaken with a vortex mixer. The resulting suspension was passed through a 0.4 m polycarbonate membrane filter 10 times and a 0.1 μm polycarbonate membrane filter 10 times at room temperature to obtain a suspension of core fine particles. 1000 L of the suspension of the obtained core microparticles and 1.333 mL of ethanol were mixed, and 480 mg of EPC and 100 mg of PEG-DSPE were mixed with 333 μL of a stirred solution of 2 mL of ethanol.

Liquid Β;

 Distilled water

<Comparative Example 1>

 Using the following coated particle manufacturing equipment, liquid Α and liquid 入 れ, put liquid A into (26), put liquid B into the following syringe pump, and put liquid B into liquid A at ImL / min for 23 minutes (23 mL) I got calo. The obtained suspension of coated fine particles was subjected to ultracentrifugation (1 hour, 110,000 X g, 25 ° C). The precipitate after ultracentrifugation was diluted with PBS, and the total lipid concentration after dilution was adjusted to 30 mg / mL to obtain a preparation.

Coated particle manufacturing equipment;

 The coated fine particle manufacturing apparatus shown in FIG. 9 was used. (27) and (28) were used as a 20 mL syringe (manufactured by Terumo) loaded in a syringe pump (Terfusion syringe pump STC-525, manufactured by Terumo). Put a rotor and framed it with a stirrer.

Liquid A;

9 mL of distilled water was added to 30 mg of FD, 180 mg of DOTAP and 72 mg of PEG-DSPE, and the mixture was shaken with a vortex mixer. The resulting suspension was passed through a 0.4 m polycarbonate membrane filter 10 times and a 0.1 μm polycarbonate membrane filter 10 times at room temperature. 750 L of the obtained suspension was placed in a glass container having a volume of about 30 mL, and a rotor was put therein and mixed with 1 mL of ethanol while stirring with a stirrer to obtain a suspension of core fine particles. Obtained To the suspension of the obtained core fine particles, ethanol (5 mL) was added to 1200 mg of EPC and 250 mg of PEG-DSPE, and 250 μL of a solution obtained by stirring was mixed with kneader.

 Liquid Β;

 Distilled water

[0094] <Test Example 1>

 For each of the preparations obtained in Examples 18 and Comparative Example 1, the average particle diameter of the coated fine particles was measured using a dynamic light scattering (DLS) measuring device (A model ELS-800, Otsuka Electronics). The results are shown in Table 1.

[Table 1] Average particle size (nm)

 Example 1

 Example 2

 Example 3

 Example 4

 Example 5

 Example 6

 Example 7

 Example 8

Comparative Example 1

[0096] <Test Example 2>

 For each of the preparations obtained in Examples 18 and Comparative Example 1, the encapsulation rate of FD in the coated fine particles was determined as follows.

30 L of each preparation was taken, 2970 L of purified water was added to each, and the mixture was stirred and ultracentrifuged to obtain a supernatant. Dilute each of the above preparations and each of the above supernatants 1000-fold, add 50 μL of 10 w / v% Triton X-100 (TritonX-100) and 400 μL of PBS to each of the diluted solutions, and vortex mixer. With stirring. 100 L of the solution was placed in a 96-well microplate, and the fluorescence intensity at an excitation wavelength of 485 nm and a fluorescence wavelength of 530 nm was measured using a fluorescent plate reader (ARVOsx-4, manufactured by Wallac). On the other hand, the fluorescence intensity of each FD aqueous solution at 1, 0.5, and 0.25 g / mL was measured, and a calibration curve was obtained. The FD concentration in each preparation and the FD concentration in each supernatant after ultracentrifugation were determined, and the encapsulation rate of FD in the coated fine particles was calculated by the following equation (1). Table 2 shows the results.

[Equation 1] Inclusion rate (!) = (Α ₁ -Β ₁ Χ100) / Α _α Χ100 (1)

 : FD concentration in drug product (g / mL)

= FD concentration in test solution prepared from drug product ( _g / mL) X1000X10

: FD concentration in the supernatant after ultracentrifugation _(w g / raL)

= FD concentration in test solution prepared from supernatant after ultracentrifugation ( _wg / inL) X 1000X10

[Table 2] Inclusion rate (%)

 Example 1

 Example 2

 Example 3

 Example 4

 Example 5

 Example 6

 Example 7

 Example 8

Comparative Example 1

[0099] <Test Example 3>

 The recovery rate for the preparation amount of FD and EPC in each preparation obtained in Example 18 and Comparative Example 1 was determined as follows.

 Each preparation was diluted 1000-fold, and 50 μL of each diluted solution was added with 50 μL of 10% w / v Triton X-100 and 400 μL of PBS, respectively, and stirred with a vortex mixer. Transfer 100 μL of the mixture to a 96-well microplate and use a fluorescence plate reader (Wallac, ARVOsx-4) to measure the fluorescence intensity at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. Was measured. On the other hand, the fluorescence intensity of each FD aqueous solution at 1, 0.5 and 0.25 g / mL was measured, and a calibration curve was obtained. The FD concentration in each preparation was determined from the calibration curve force. On the other hand, the EPC concentration in each preparation was determined by measuring using a phospholipid C-Test Co. (Wako Pure Chemical Industries, Ltd.). The FD recovery rate and EPC recovery rate for each formulation were calculated by the following formula (2). Table 3 shows the results.

[0100] [number 2] FD recovery in drug product (%) = A ₂ / B ₂ X 100 _(}

EPC recovery in drug product (»= C ₂ ZD ₂ X 100

A 2: FD concentration in drug product (μg / mL)

= Concentration of FD in test solution prepared from drug product (g / niL) X1000X10 B 2: Theoretical FD concentration in drug product with recovery rate of 100% (/ i _g / mL)

C ₂ : EPC concentration in drug product (mg / mL)

D ₂ : Theoretical EPC concentration in drug product with 100% recovery ( _mg / mL)

[0101] [Table 3]

FD recovery rate (%) EPC recovery rate (%)

 Example 1 4 5.6 4 3.2

 Example 2 6 2.9 6 2.6

 Example 3 6 4.6 5 5.8

 Example 4 5 6.5 6 2.8

 Example 5 6 1.8 6 4.2

 Example 6 5 8. 6 5 4.6

 Example 7 5 8.5 5 5 3.8

 Example 8 6 9.2 79.0

 Comparative Example 1 5 8. 7 6 4. 7

[0102] <Test Example 4>

 The stability of each of the preparations obtained in Examples 18 and Comparative Example 1 in fetal bovine serum (FBS) was examined.

 Each preparation Uko FBS 2970 L was mixed with syrup. Immediately after mixing and after standing at 37 ° C for 3 hours, 500 L of each sample was taken and subjected to gel filtration chromatography (Gel Permeation Chromatography, GPC) to collect the fractions (100 drops, 10 tubes). Each fraction is shaken and stirred with a vortex mixer to make a sample.

10 w / v% TritonX-100 (50 μL) and PBS (400 μL) were added, and the mixture was stirred with a vortex mixer. 100 L of the solution was placed in a 96-well microplate, and the fluorescence intensity at an excitation wavelength of 485 nm and an emission wavelength of 530 nm was measured using a fluorescent plate reader (Wallac, ARVOsx-4). . The FD recovery rate after 0 hour and 3 hours was calculated by the following equation (3). Table 4 shows the results.

[0103] [Number 3]

FD recovery rate after 0 hour (%) = 8 /: 6 ₃ ><1 0 0

FD recovery rate after 3 hours (%) = Ji _{3/0 3 X 1 0 0} '

 4 = ^ (each fraction * light intensity coated fine particle fraction. After time) (each fraction volume) >> coated fine particle fraction. After hours) )}

)} = ∑ {[Each Fraction Yodo Mitsutaka full picture. Time _ί (Each fraction volume

)}

 c = i (each nonction light aae painting);

A = ∑ {(Each fraction fluorescence intensity _Total fraction time _ί (Each fraction volume) _m , _m) )}

[0104] [Table 4]

 0 hours later 3 hours later

 Recovery rate (%) Recovery rate (%)

 Example 17 99.5 76.3

 Example 2 7 7 .3 7 4 .2

 Example 3 7 2 .0 70.7

 Example 4 7 1. 9 6 8.5

 Example 5 7 2 .3 6 7 .9

 Example 6 7 5 .4 7 1.7

 Example 7 7 2 .1 7 2 .0

 Example 8 7 2 .6 7 1.5

 Comparative Example 16 6. 8 6 3.6

 [0105] As is clear from Tables 1 and 2, the coated microparticles in each of the preparations obtained in Examples 18 to 18 and Comparative Example 1 had the same average particle diameter and the same encapsulation rate, and apparently As a result, coated fine particles of similar quality were obtained. In addition, as can be seen from Table 3, the recovery rates of FD and EPC were the same, and coated fine particles were obtained with the same efficiency. On the other hand, as can be seen from Table 4, in each of the preparations obtained in Examples 18 to 18, the recovery of FD in PBS was higher than in the preparation obtained in Comparative Example 1. Good stability at That is, despite the fact that the coated fine particles can be easily prepared by the kit of the present invention, the coated fine particles having the same size and the drug encapsulation rate as those of the conventional method can be made to contain the drug more firmly with the same efficiency. It was also possible to manufacture it in a state where it was left out.

Industrial applicability According to the present invention, a kit for preparing coated microparticles in which core microparticles are coated with a coating layer is provided.

Claims

The scope of the claims  [1] A liquid containing a polar organic solvent in which core fine particles are dispersed and a coating layer component constituting a coating layer for coating the core fine particles is dissolved (liquid, liquid  A solvent that can be mixed with Liquid A and contains no polar organic solvent or contains a lower percentage of polar organic solvent than Liquid A (Liquid B),  And a preparation unit for preparing coated microparticles including a device provided with means for mixing liquid A and liquid B.  [2] An apparatus provided with a means for mixing the liquid A and the liquid B is a means for storing the liquid A, a means for storing the liquid B, an in-line mixing means, and a means for storing the liquid A. 2. The kit for preparing coated microparticles according to claim 1, wherein the kit is provided with a liquid sending means to an inlet of the in-line mixing means and a liquid sending means from a means for containing the liquid B to an inlet of the in-line mixing means. .  [3] An instrument having a means for mixing the liquid A and the liquid B A syringe for storing the liquid A, a syringe for storing the liquid B, and a syringe for storing the liquid A and the liquid B 2. The kit for preparing coated microparticles according to claim 1, wherein the kit is provided with in-line mixing means having two inlets to which syringes can be respectively connected.  [4] The device provided with a means for mixing the liquid A and the liquid B is a multi-chamber syringe having chambers for storing the liquid A and the liquid B in parallel, respectively. 2. The kit for preparing coated microparticles according to claim 1, wherein the kit is a syringe.  [5] The in-line mixing means according to any one of claims 2 to 4, wherein the kit is a in-line mixing means connected or built in with a static mixer.  [6] a device provided with a means for mixing liquid A and liquid B, means for containing liquid A, means for containing liquid B, and means for containing liquid B 2. The kit for preparing a coated fine particle according to claim 1, wherein the kit comprises means for adding the liquid B to the liquid A in the means. [7] An instrument having a means for mixing the liquid A and the liquid B. A syringe for storing the liquid A and a syringe for storing the liquid B. The two syringes are connected to connect the liquid B to the liquid A. 2. The kit for preparing coated microparticles according to claim 1, which can be dried. [8] An apparatus provided with a means for mixing the liquid A and the liquid B is a multi-chamber syringe having chambers for respectively accommodating the liquid A and the liquid B, and the syringe is configured to transfer the liquid B to the liquid A. 2. The kit for preparing coated microparticles according to claim 1, wherein the kit is a syringe that can be dried.  [9] The kit for preparing coated microparticles according to any one of claims 2 to 8, wherein the device provided with a means for mixing the liquid A and the liquid B is a device provided with a transfer needle or a co-injection connector at an outlet. .  [10] The kit for preparing coated microparticles according to any one of claims 19 to 19, wherein the coating layer component is at least one substance selected from lipids, surfactants and polymers.  [11] The kit for use in preparing coated microparticles according to any one of claims 110, wherein the coating layer is a lipid membrane.  [12] The use of the coated microparticle according to any one of claims 11 to 11, wherein the core microparticle is a microparticle comprising a drug, a lipid assembly, a ribosome, an emulsion microparticle, a polymer, a metal colloid or a microparticle preparation. Preparation kit. 13. The microparticle of claim 11, wherein the core microparticle is a microparticle comprising a complex composed of two or more drugs, lipid aggregates, ribosomes, emulsion microparticles, polymers, metal colloids, or microparticle preparations. 2. A kit for preparing the coated fine particles according to the item 1 above. 14. The kit for preparing coated microparticles according to claim 11, wherein the core microparticles are microparticles comprising a complex of a drug and a ribosome. [15] The kit for preparing coated microparticles according to any one of claims 114, wherein the polar organic solvent is at least one selected from alcohols, glycols and polyalkylene glycols.  [16] Coated microparticles which can be prepared by the kit for preparing coated microparticles according to claim 11 when used.