WO2013161778A1 - 薬剤ナノ粒子を分散した水分散液の製造法およびその利用 - Google Patents
薬剤ナノ粒子を分散した水分散液の製造法およびその利用 Download PDFInfo
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- WO2013161778A1 WO2013161778A1 PCT/JP2013/061819 JP2013061819W WO2013161778A1 WO 2013161778 A1 WO2013161778 A1 WO 2013161778A1 JP 2013061819 W JP2013061819 W JP 2013061819W WO 2013161778 A1 WO2013161778 A1 WO 2013161778A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/27—Esters, e.g. nitroglycerine, selenocyanates of carbamic or thiocarbamic acids, meprobamate, carbachol, neostigmine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
- A61K51/082—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins the peptide being a RGD-containing peptide
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0048—Eye, e.g. artificial tears
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Definitions
- the present invention relates to a method for producing an aqueous dispersion in which drug nanoparticles are dispersed and use thereof.
- drug nanocrystals drug nanocrystals
- particles of drugs particularly poorly water-soluble drugs
- Drugs are expected to have improved penetration into cells and / or tissues due to their nanosize.
- the bulk powder of the drug is sparingly water-soluble, the water-solubility of the drug may be significantly improved by nanocrystallization.
- drug nanocrystals are rapidly studied in the scientific and pharmaceutical fields as an excellent next-generation pharmaceutical form.
- Non-Patent Document 1 a technique in which drug bulk powder is refined in a top-down manner by a homogenizer, laser ablation, milling, etc.
- Patent Documents 1 and 2 Techniques such as precipitation method, vapor phase deposition method, emulsion method, spray drying method, freezing method, etc. for producing drug nanocrystals from an aggregate of drug molecules in a bottom-up manner
- Non-Patent Document 1 Techniques such as precipitation method, vapor phase deposition method, emulsion method, spray drying method, freezing method, etc. for producing drug nanocrystals from an aggregate of drug molecules in a bottom-up manner
- the present invention has been completed to solve the above problems, and more specifically, to provide an aqueous composition in which nanoparticles are dispersed, which can be stably used as an aqueous dispersion preparation. .
- the present inventors have made extensive studies and, as a result, prepared using a mixed solution of an organic solvent and water containing an active ingredient as a drug and an ointment base.
- the drug nanoparticles it was found that the above-mentioned nanocrystals of the poorly water-soluble drug can be dispersed in water, and in the obtained nanoparticle aqueous dispersion, the crystal growth of the nanocrystals was suppressed.
- the original characteristics of the ointment base contained in nanoparticles (improving the retention of the drug in the affected area, maintaining the long-term sustained release of the drug, etc.)
- the present invention has been completed by finding that it is maintained.
- the method of the present invention comprises a first solution using an organic solvent as a solvent and a second solution using water as a solvent.
- a step of freeze-drying a sample obtained by freezing the mixed solution with the step, wherein the mixed solution contains an active ingredient and an ointment base, and a step of dispersing the freeze-dried sample in water It is characterized by that.
- the nanoparticles containing the active ingredient are uniformly dispersed, and no crystal growth occurs even after long-term storage. .
- the method of the present invention may further include a step of confirming that crystal growth of particles dispersed in the aqueous dispersion has not occurred.
- the step of confirming may be a step of confirming that no precipitation has occurred in the aqueous dispersion.
- the method of the present invention can also be a method for suppressing the growth of nanocrystals of the active ingredient. This is presumably because the surface of the active ingredient was coated with the ointment base by rapidly freezing and freeze-drying the mixture of the organic solvent and water containing the active ingredient and the ointment base.
- the surface of the active ingredient is successfully coated with the ointment base, and as a result, the growth of nanocrystals of the active ingredient is suppressed.
- the accuracy of the coating may not be constant.
- the method of the present invention may further include, as necessary, a step of selecting a dispersion in which nanoparticle crystal growth has not occurred from the obtained plurality of aqueous dispersions. It may be a step of selecting an aqueous dispersion in which no precipitation has occurred from a plurality of aqueous dispersions, or a step of collecting a suspension fraction (a step of removing the generated precipitate).
- the nanoparticle aqueous dispersion of the present invention is characterized by being manufactured according to the method described above.
- the composition of the present invention is characterized by containing the above aqueous dispersion of nanoparticles.
- the composition of the present invention may be an eye drop or an external preparation for skin as a substitute for an ointment.
- the composition of the present invention contains a nano-sized drug, it may be used to deliver the active ingredient of the drug to the deep tissue, for example, to treat a disease in the deep eye Is preferably used for this purpose.
- the crystal growth of nanocrystals can be suppressed without impairing the characteristics of the ointment base that imparts sustained release and retention to an ointment (ointment). Furthermore, if the present invention is used, a poorly water-soluble drug can be successfully dispersed in water without causing precipitation over a long period of time.
- the present invention it becomes possible to locally apply the drug at the nano level as an aqueous dispersion preparation, and the inconvenience (inconvenience or discomfort) of the ointment due to the wide application around the affected area is reduced.
- the present invention provides a method for producing a nanoparticle aqueous dispersion.
- the present invention provides a method for producing an aqueous dispersion of drug nanoparticles in which a poorly water-soluble or water-insoluble drug (or an active ingredient thereof) is successfully dispersed.
- a mixture of an organic solvent and water containing an ointment base together with a target active ingredient for example, a poorly water-soluble or water-insoluble drug
- a target active ingredient for example, a poorly water-soluble or water-insoluble drug
- the production method of the present invention freezes a sample obtained by freezing a mixed solution of a first solution using an organic solvent as a solvent and a second solution using water as a solvent in order to produce a nanoparticle aqueous dispersion.
- the active ingredient (poorly water-soluble or water-insoluble drug) suitably used in the present invention is not particularly limited, and for example, steroids (dexamethasone and fluorometholone, and triamcinolone, which are used in suspension type eye drops described later, Inhibitors that specifically act on triamcinolone acetonide, betamethasone, hydrocortisone, prednisolone, cortisone, prednisolone acetate, methylprednisolone, methylprednisolone succinate, betamethasone valerate, cortisol succinate) and calpain (calcium-dependent cysteine protease) Calpain inhibitors that function as neuroprotective agents by inhibiting cell apoptosis (calpain inhibitor I, calpeptin, calpain inhibitor II, calpain inhibitor -III, Calpain inhibitor IV, Calpain inhibitor IV-2, Calpain inhibitor V, Calpain inhibitor VI, Calpain inhibitor VII, Cal
- the method of the present invention further includes a step of confirming that nanoparticle crystal growth has not occurred, and a dispersion liquid in which nanoparticle crystal growth has not occurred. More preferably, the method further includes the step of selecting.
- Non-Patent Documents 1 and 2 nanoparticles of a drug incorporated in a matrix are obtained by freezing and then lyophilizing a mixture of t-butyl alcohol and water in which the drug and matrix are dissolved.
- Non-Patent Documents 1 and 2 neither describe nor suggest that drug nanoparticles are produced using an ointment base.
- mannitol is used as a matrix in Non-Patent Documents 1 and 2 as a carrier for facilitating crystallization during lyophilization, but mannitol is not only an ointment base but is common in terms of structure between the two. There is no point.
- Non-Patent Document 1 states that when dispersed in a solvent, drug nanocrystals can easily form or dissolve aggregates, and the formation of aggregates can be suppressed by adding a surfactant. It is described that surfactants partially dissolve the drug, leading to the formation of non-optimal crystal size. These descriptions indicate that the crystal growth of the drug in water such as Ostwald ripening is difficult, and that it is difficult to redisperse the drug nanoparticles in water. However, Non-Patent Documents 1 and 2 do not disclose or suggest a technique for overcoming these disadvantages.
- Non-Patent Documents 1 and 2 are difficult to re-disperse in water, and crystal growth of drug nanoparticles in an aqueous dispersion cannot be suppressed. .
- Patent Document 1 an emulsifier is contained in an aqueous phase, a non-polymeric hydrophobic organic compound insoluble in water is contained in an organic phase together with a drug, and the aqueous phase and the organic phase are mixed using a high-pressure homogenizer. Emulsion is prepared, and drug nanoparticles are obtained by removing the contained organic solvent under vacuum. In Patent Document 2, drug nanoparticles are obtained by milling a drug having a fatty acid ester adsorbed on the surface in a dispersion medium.
- Patent Documents 1 and 2 neither describe nor suggest that drug nanoparticles are produced using an ointment base or that lyophilization is used in the production of drug nanoparticles.
- the crystal growth of the dispersed particles preferably does not occur for at least one week, and more preferably does not occur for one month or more.
- the crystal growth of the dispersed particles is It is preferable that it does not occur, it is more preferable that it does not occur for at least one week even in a high temperature environment, and it is even more preferable that it does not occur for one month or more. That is, the step of confirming is preferably performed one week or more after the step of dispersing, and more preferably performed after one month, two months, three months, six months, twelve months or more. In some cases, the process may be performed in the harsh environment.
- Examples of the organic solvent used in the first solution include cyclohexane, benzene, 1,4-dioxane, ⁇ , ⁇ , ⁇ -trifluorotoluene, t-butyl alcohol, t-amyl alcohol, ethyl isothiocyanate, 4- Xylene, 2-xylene, 2-chloro- ⁇ , ⁇ , ⁇ -trifluorotoluene, 4-chlorotoluene, cyclohexanol, ethanol, acetone, methanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, 2-methyl-1-propanol, 2-methyl-2-propanol, chloroform, dibromomethane, butyl chloride, dichloromethane, dimethoxymethane, tetrahydrofuran, diethyl ether, ethylene glycol, dimethyl ether, ethylene glycol diethyl ether
- the second solution may be water or water containing a dispersant as required.
- the crystal growth of the nanoparticles may not be successfully suppressed in the obtained dispersion, but as described above, the nanoparticles
- a dispersion can be produced.
- the mixed solution of the first solution and the second solution preferably has a mixing ratio of 10,000: 1 to 1: 10000. More preferably, the ratio is 10: 1 to 1:10.
- the step of confirming may be a step of confirming that no precipitation has occurred in the aqueous dispersion
- the step of selecting may be a step of selecting an aqueous dispersion in which no precipitation has occurred, or a suspension. It may be a step of collecting the turbid liquid fraction (step of removing the generated precipitate).
- the ointment base used in the method of the present invention is not particularly limited as long as it is a known base used in producing ointments.
- lanolin petrolatum, beeswax, phenol / zinc flower liniment, cocoa butter, witepsol
- Suitable examples include glycerogelatin, liquid paraffin, hard fat, macrogol, hydrocarbon gel ointment base (for example, a mixture of liquid paraffin and polyethylene such as plastibase (Taisho Toyama Pharmaceutical Co., Ltd.)), lanolin or its Derivatives are more preferred.
- Lanolin which is a sheep oil produced as a by-product when separating wool from sheep wool, is a mixture of aliphatic alcohols, cholesterols, tempen alcohols, fatty acids, and the like. Lanolin is paste-like at room temperature, has excellent water retention, and can be mixed with twice the amount of water of its own weight, but is essentially insoluble in water. Alcohol (lauric alcohol) or acid (lauric fatty acid) obtained by hydrolysis and fractional purification of such lanolin is also a lanolin derivative.
- a product obtained by subjecting lanolin to a chemical reaction such as acetylation, alkoxylation, sulfonation, hydrogenation, transesterification and reduction is also a lanolin derivative, and a metal salt of lauric fatty acid is a lanolin derivative.
- the lanolin or a derivative thereof used in the method of the present invention may be a mixture of lanolin derivatives, and is most preferably purified lanolin.
- an ointment base comprising an aqueous ointment base such as macrogol ointment, an emulsion ointment base, or a mixed base thereof may be used.
- polysorbate 80 Teweens
- Emulsifiers such as triethanolamine, gum arabic, tragacanth, sodium lauryl sulfate, polyoxyl 40 stearate, sorbitan monofatty acid ester (Spans), and other surfactants may be blended.
- the method of the present invention may further include a step of preparing the first solution by dissolving the active ingredient and the ointment base in an organic solvent, and a step of preparing the second solution by dissolving the dispersant in water.
- a step of preparing a mixed solution of the first solution and the second solution and a step of freezing the mixed solution of the first solution and the second solution may be further included.
- quick freezing is desired, for example, in an environment where the mixed solution freezes, for example, in an environment of 0 ° C.
- a refrigerator use of a freezer, use of a deep freezer, in a cooling solvent environment, in a liquid helium environment It is preferably carried out in a dry nitrogen environment, in a liquid nitrogen environment, etc., but most preferably in a liquid nitrogen environment.
- nanoparticle is intended to be a particle having a particle size of nanometer, and the nanoparticle in the present invention preferably has an average particle size of about 500 nm, The size may be less than 220 nm. Nanoparticles having such a particle size can be obtained by filtering using a 0.22 ⁇ m filter.
- the average particle size of the nanoparticles in the present invention is more preferably about 10 to 200 nm, more preferably about 20 to 180 nm, still more preferably about 25 to 150 nm, and most preferably about 25 to 100 nm. .
- the method of the present invention preferably further includes a step of filtering the dispersion obtained in the step of dispersing. This step is preferably used for removing particles exceeding the nanosize from the aqueous dispersion, but may be used as a step for confirming that no crystal growth of the nanoparticles occurs.
- the solubility of a drug can be improved with an increase in specific surface area due to nano-ization.
- a poorly water-soluble drug it becomes possible to disperse the poorly water-soluble drug necessary for administration to a living body.
- the “active ingredient” is not limited to a specific drug, as long as it is a drug used in an aspect where the effect of the nanoparticle aqueous dispersion of the present invention is expected.
- prodrugs as disclosed in WO 2010/053101 are also preferably used as active ingredients applied to the present invention.
- the method for producing a pharmaceutical nanoparticle aqueous dispersion according to the present invention includes at least a step of freeze-drying a sample obtained by freezing a mixed solution of the first solution and the second solution, and using the freeze-dried sample in water. It can be said that the process of dispersing may be included. That is, it should be noted that methods in which the drug (active ingredient), the ointment base, and the organic solvent are different from the examples described later are also included in the technical scope of the present invention.
- an object of the present invention is to provide an aqueous dispersion of drug nanoparticles including a step of freeze-drying a sample obtained by freezing a mixed solution of a first solution and a second solution, and a step of dispersing the freeze-dried sample in water. It is to provide a manufacturing method, and not to individual drugs (active ingredients), ointment bases and organic solvents specifically described herein.
- the present invention also provides a nanoparticle aqueous dispersion produced by the method described above.
- particles having a particle size of nanometer size, in which an ointment base is contained together with a target active ingredient (a poorly water-soluble or water-insoluble drug) are dispersed in water.
- a target active ingredient a poorly water-soluble or water-insoluble drug
- the ointment base contained in the nanoparticles dispersed in the aqueous nanoparticle dispersion of the present invention is not particularly limited as long as it is described above.
- the nanocrystal aqueous dispersion prepared according to the conventional method is a heterogeneous system having a spread in particle distribution, which is equivalent to a mode in which non-nanocrystals are suspended, and the advantages as nanocrystals are lost. Yes.
- nanoparticle crystal growth does not occur. This is considered to be because part or all of the surface of the active ingredient was coated with the ointment base. That is, in the nanoparticles dispersed in the aqueous nanoparticle dispersion of the present invention, the surface of the active ingredient (poorly water-soluble or water-insoluble drug) is coated with the ointment base.
- nanoparticles dispersed in the aqueous nanoparticle dispersion of the present invention are stored in a high-temperature environment (40 ° C. or higher, 50 ° C. or higher, or 60 ° C. or higher), at least one week or longer, Preferably no crystal growth occurs over a period of 1 month or longer, more preferably 2 months, 3 months, 6 months, 12 months or longer. That is, in the nanoparticle aqueous dispersion of the present invention, no precipitation system is formed over a period of at least 1 week, preferably 1 month or more, more preferably 2 months, 3 months, 6 months, 12 months or more.
- the nanoparticles dispersed in the aqueous nanoparticle dispersion of the present invention preferably have an average particle size of about 500 nm, but may have a size of less than 220 nm. Nanoparticles having such a particle size can be obtained by filtering using a 0.22 ⁇ m filter.
- the average particle size of the nanoparticles in the present invention is more preferably about 10 to 200 nm, more preferably about 20 to 180 nm, still more preferably about 25 to 150 nm, and most preferably about 25 to 100 nm. .
- the particle size of the nanoparticles in the present invention is within the above-mentioned numerical range, the migration of the drug contained in the nanoparticles is remarkably increased, and the efficiency of delivering the active ingredient of the drug to a deep tissue region is greatly improved. Is expected to do.
- the penetration of a drug into the deep part of the eye increases, so that it is possible to treat an intractable disease in the deep part of the eye (such as the retina), where drug treatment has been difficult.
- the maximum pharmacological effect by the minimum administration of the drug can be expected by increasing the transferability of the drug.
- the present invention provides a composition containing a nanoparticle aqueous dispersion.
- the composition of the present invention can be used for various applications due to the properties of the nanoparticle aqueous dispersion.
- Ointment substitute is a semisolid external preparation in which an active ingredient is dispersed in a base such as petrolatum, and is applied to the affected area to treat skin diseases and the like.
- Bases used in ointments include hydrophobic bases (oil bases) such as petrolatum, hydrophilic bases (emulsifying bases, water-soluble bases, suspending bases). Agent), hydrophobic base, paste (liniment), mud (pasta), plaster, lotion, spray, etc., and the usage form of ointment is basically an application type such as cream, gel, lotion, etc. .
- Ointments can retain active ingredients on the skin for a long time by adhering to the skin due to the nature of the ointment base, and are used in skin external preparations, oral ointments, eye ointments and the like.
- the ointment has the advantage of long-term and stable sustained release of the drug because the state applied to the diseased part is maintained for a long time.
- it is not easy to clean the ointment.
- ointments for eye diseases eye ointments
- eye ointments are inconvenient due to suspension due to the application of cream, and have low patient compliance.
- a target diseased part is limited to skin, eyes, an oral cavity, etc., and a patient cannot apply easily to the diseased part in a body.
- the application range is as wide as several mm 2 to several cm 2 , the drug is often applied to healthy tissues other than the diseased part.
- the nanoparticles dispersed in the aqueous nanoparticle dispersion of the present invention contain an active ingredient (poorly water-soluble or water-insoluble drug) and an ointment base, and the surface of the active ingredient Part or all is coated with an ointment base.
- the nanoparticle aqueous dispersion not only the crystal growth of the nanocrystals is suppressed, but also the nanoparticle aqueous dispersion has the original characteristics of the ointment base contained in the nanoparticle (the drug on the affected area). Such as improving the retention and maintaining the long-term sustained release of the drug).
- the composition of the present invention can provide drug nanoparticles in an aqueous dispersion (that is, a liquid preparation), it realizes a dosage form (oral administration, injection, eye drops, etc.) that has been difficult with conventional ointment preparations. To do.
- the composition of the present invention is used as an ointment substitute, and more particularly, the composition of the present embodiment is an eye drop.
- the composition of the present invention was provided in the form of an aqueous dispersion, and droplet administration was realized. Thereby, since it is not necessary to apply thickly unlike the conventional ointment formulation, there is no sticky discomfort, and poor visibility caused by application like the conventional ointment formulation does not occur.
- suspension type eye drops in which a poorly water-soluble drug is suspended / dispersed in water are commercially available.
- Steroid is mentioned as a typical example of the chemical
- steroids are poorly water-soluble, they form a large particle mass of several micrometers or more in water and are deposited on the bottom of the eye drop container.
- Such a suspension-type drug has a large particle size, and therefore has low drug solubility and transferability into the eye (5% or less).
- the composition of the present invention contains a nano-level drug, the penetration of the drug into cells / tissues is increased, and deep eye inflammation and posterior ocular retina, which are difficult to achieve with conventional ophthalmic solution administration.
- composition of the present invention is expected to have long-term retention and sustained release derived from the ointment base.
- the composition of this embodiment is an external preparation for skin.
- the preparation is provided in nano size, and the permeability to tissues and cells is improved.
- the ointment base exerts its function, whereby the retention of the drug in the affected area can be improved and the long-term sustained release of the drug can be maintained.
- the drugs used in the present invention are mainly poorly water-soluble drugs and are nanosized.
- the preparation of the composition of the present invention is provided in a nano size, and the permeability to tissues and cells is improved.
- the composition of the present invention is an extremely effective dosage form for DDS (drug delivery system) that can be expected to have the maximum effect due to the minimum administration of the drug, and is expected to have excellent pharmacological effects and improved patient compliance. Is done. That is, the composition of the present invention is used in an aspect where it is necessary to deliver an active ingredient of a drug from an applied site (tissue surface) to a deep tissue part.
- the composition of the present invention When used as the above-mentioned eye drops, the composition of the present invention is used for treating diseases in the deep part of the eye (such as the retina).
- diseases in the deep part of the eye such as the retina.
- the composition of the present invention provides a new dosage form, it can be expected to be used in many medical fields and have outstanding economic effects.
- the present invention further provides a method of inhibiting nanocrystal growth. Suppressing the crystal growth of drug nanocrystals is an extremely important issue in the formulation of nanocrystal preparations that will be developed in the future.
- the present invention provides a method for inhibiting crystal growth by coating the surface of drug nanocrystals using an ointment base.
- the drug nanocrystal to which the present invention is applied is provided in a new formulation form that retains the characteristics (sustained release and retention) of the coated ointment formulation.
- the method of the present invention includes a step of lyophilizing a sample obtained by freezing a mixed solution of a first solution containing an organic solvent as a solvent and a second solution containing water as a solvent. And an ointment base.
- a target active ingredient for example, a poorly water-soluble or water-insoluble drug
- Drug nanocrystals are obtained by freezing a mixture of organic solvent and water (containing the drug) and then lyophilizing the frozen sample, but by using the method of the present invention, Crystal growth is suppressed. This is because the surface of the active ingredient is coated with the ointment base by rapidly freezing and lyophilizing the mixture of the first solution and the second solution (containing the drug and the ointment base). This is probably because Thus, if the present invention is used, growth of nanocrystals can be suppressed.
- this method refer to the description of “1: Method for producing nanoparticle aqueous dispersion” as appropriate.
- the method of the present invention preferably further includes a step of confirming that no crystal growth of the nanoparticles has occurred, and a step of dispersing the lyophilized sample in water, More preferably, the method further includes a step of selecting a dispersion in which nanoparticle crystal growth has not occurred.
- a uniform dispersion system is maintained because crystal growth of nanoparticles does not occur, and as a result, a precipitation system is not formed.
- the step of confirming may be a step of confirming that precipitation has not occurred in the aqueous dispersion, and the step of selecting selects an aqueous dispersion in which precipitation has not occurred from a plurality of aqueous dispersions. It may be a step of selecting or a step of collecting a suspension fraction (a step of removing a generated precipitate).
- Dexamethasone, lanolin, t-butyl alcohol, polyvinylpyrrolidone, polysorbate 80 were obtained from Wako Pure Chemical Industries, Ltd., polyoxyethylene (200) polyoxypropylene glycol (70) was obtained from Nippon Oil and Fatty, and hydroxypropylmethylcellulose. Was obtained from Shin-Etsu Chemical.
- the obtained dispersion was passed through a filter having a pore size of 0.22 ⁇ m to obtain an aqueous dispersion of dexamethasone nanoparticles. Further, according to the same procedure, an aqueous dispersion of fluorometholone nanoparticles was obtained.
- FIG. 1 shows an electron microscope image of dexamethasone nanocrystals in the aqueous dispersion of dexamethasone nanoparticles.
- an aqueous dispersion of dexamethasone was prepared by ultrasonic suspension according to a conventional procedure, and dexamethasone nanocrystals in the aqueous dispersion were observed with an electron microscope (FIG. 2).
- the prior art procedure is as follows: lanolin (60 mg), polyoxyethylene (200) polyoxypropylene glycol (70) (60 mg), polyvinylpyrrolidone (20 mg), hydroxypropyl methylcellulose (3 mg), and polysorbate 80. (5 ul) is mixed with 10 mL of purified water to prepare a mixed solution. Subsequently, dexamethasone (30 mg) is mixed into this mixed solution, and ultrasonic irradiation is performed with an ultrasonic generator (BRASON 2510) for 60 minutes. An aqueous dispersion of dexamethasone by suspension was prepared.
- the target crystal sample was isolated by vacuum filtration using an isopore membrane filter (MILLIPORE, filter code: VMTP) having a pore size of 0.05 ⁇ m. Collected on a membrane filter.
- An isopore membrane filter carrying a crystal sample is attached to a scanning electron microscope (SEM) (manufactured by JEOL: JSM-6510LA) using a conductive carbon tape, and sputtering (manufactured by JEOL: JFC). -1600) and applying a platinum coating to the surface of the crystal sample, the crystal sample was observed with an SEM.
- SEM scanning electron microscope
- a nanocrystal group of about 100 nm was observed, and it was found that good nanoparticles could be obtained according to the method of the present invention.
- a crystal habit of several to several tens of micrometers was formed, and it was found that good nano-level particles could not be obtained according to the procedure of the prior art.
- the method of the present invention is a bottom-up method in which nanocrystals (or nanoparticles coated with a base material) are produced by a crystallization process based on a collection of molecules present in a solution by a rapid freezing process. is there.
- the conventional manufacturing method is a method in which a particle lump is crushed in a top-down manner by applying an ultrasonic impact to a large particle lump.
- ultrasonic generator used in the conventional production method, there is a limit to miniaturization of the particle diameter, and usually a group of particles having an average particle diameter of the order of ⁇ m or more is produced.
- ultrasonic treatment is also performed in the production process using the same apparatus, which is used to loosen a lump of nanoparticles produced by freeze-drying method and re-disperse in water by ultrasonic impact. Yes, not for crushing particles.
- FIG. 3 shows an electron microscopic image of the fluorometholone nanocrystals in the fluorometron nanoparticle aqueous dispersion.
- a group of nanocrystals of about 100 nm was observed, and it was found that good nanoparticles can be obtained according to the method of the present invention.
- the fluorometholone aqueous dispersion A using lanolin that is, the nanoparticle dispersion of the present invention
- lanolin that is, the nanoparticle dispersion of the present invention
- a fluorometholone aqueous dispersion B was prepared. A severe test is performed in which the dispersions A and B are allowed to stand in an oil bath at 60 ° C. for 48 hours, and the degree of crystal growth occurring before and after the severe test is observed with an electron microscope and The particle size distribution was measured by dynamic light scattering method and zeta potential measurement.
- dispersion A there was almost no change in the particle size, particle size distribution, and zeta potential before and after the severe test, but in dispersion B, nanoparticle crystal growth and particle size distribution spread after the severe test.
- the particle dispersion was unstable due to a significant decrease in zeta potential.
- An electron microscope image is shown in FIG. 5, and the particle size distribution and zeta potential are shown in FIG.
- the crystal growth inhibitory effect of the nanoparticles in the nanoparticle dispersion of the present invention was demonstrated.
- calpeptin aqueous dispersion C using lanolin namely, the nanoparticle dispersion of the present invention
- calpeptin aqueous dispersion D not using lanolin were prepared using calpeptin which is a calpain inhibitor.
- Nanoparticles with a particle size of about 100 nm in which crystal growth was suppressed were observed in Calpeptin aqueous dispersion C, but the crystal growth was not suppressed in Carpeptin aqueous dispersion D, and the average size of the fibers was less than 4 ⁇ m.
- a drug was formed. The results of these electron microscope observations, the particle size distribution measurement by the dynamic light scattering method, and the zeta potential measurement are shown in FIGS.
- Table 1 shows the results of the particle size distribution and zeta potential of the nanocrystals in the obtained aqueous dispersion of nanoparticles obtained by the dynamic light scattering method.
- the dispersed particles are small in size, the particles are uniformly dispersed and light scattering is reduced, and the zeta potential measurement shows that the negatively charged nanoparticles are stably dispersed. It was.
- FIG. 9 shows an electron microscope image of the nanocrystals in the obtained nanoparticle aqueous dispersion
- Table 1 shows the results of particle size distribution and zeta potential measurement by the dynamic light scattering method.
- FIG. 10 shows an electron microscopic image of the nanocrystal in the obtained nanoparticle aqueous dispersion.
- (a) to (d) are nanocrystal images when calpain inhibitor I, calpeptin, cyclosporin A, and 7-ethyl-10-hydroxycamptothecin are used as drugs.
- Table 2 shows the results of particle size distribution and zeta potential of the nanocrystals in the aqueous nanoparticle dispersion obtained in various drugs by the dynamic light scattering method.
- ALLN and SN38 in the table indicate calpain inhibitor I and 7-ethyl-10-hydroxycamptothecin, respectively.
- the dispersed particles are small in size, the particles are uniformly dispersed and light scattering is reduced, and the zeta potential measurement shows that the negatively charged nanoparticles are stably dispersed. It was. Further, it has been found that the use of the present invention makes it possible to form not only low molecular weight compounds but also peptides (calpeptin) or oil droplets (clofibrate) drugs into nanoparticles.
- the nanoparticle aqueous dispersion of the present invention can release the target compound percutaneously into the body.
- IRBP retina-specific antigen S antigen and interphotoreceptor reretinoid binding ⁇ protein
- mice were drunk anesthetized and subcutaneously injected with 200 ⁇ L of a 5 mg / mL peptide solution in which complete Freund's adjuvant (CFA) containing tuberculosis killed bacteria and a synthetic peptide (N-terminal 15 amino acids of IRBP) were mixed thoroughly. .
- CFA complete Freund's adjuvant
- tuberculosis killed bacteria and a synthetic peptide (N-terminal 15 amino acids of IRBP) were mixed thoroughly.
- CFA complete Freund's adjuvant
- IRBP synthetic peptide
- the eye drop of the present invention can be expected to have an excellent anti-inflammatory effect against posterior ocular retinal uveitis.
- the concentration of fluorometholone in the aqueous humor was determined to be about 235 ng / mL, which is a system using commercially available eye drops reported in the literature (New Ophthalmology 7, 1051-53, 1990). Intraocular transfer was about 7 to 8 times higher than about 30 ng / mL. Thus, it was shown that the eye drop of the present invention has higher tissue permeability (intraocular transferability) than conventional eye drops.
- Eye drops were prepared using the aqueous nanoparticle dispersion of the present invention, and the therapeutic effect of retinal degenerative diseases was verified. Instillation was started twice a day from the 5th day of birth to mice (C3H / HeNCrlCrlj), which used calpain inhibitors (calpain inhibitor I and calpeptin) as drugs, and showed congenital retinal degeneration. As controls, suspension-type calpain inhibitor ophthalmic solutions having a coarse particle size (that is, not nanoparticles) and ophthalmic solutions containing only a base not containing calpain inhibitors were used.
- the C3H / HeNCrlCrlj mouse is a mouse having a genetic mutation in phosphodiesterase (PDE) 6B, in which retinal degeneration begins immediately after birth and significant retinal pigment epithelial cells and photoreceptor cells fall into apoptosis at 2 weeks of age. It is considered a model animal for retinitis pigmentosa.
- PDE phosphodiesterase
- assessing the survival of photoreceptor cells assessing the cell density and layer structure of the outer granule layer where the nucleus of the photoreceptor cell is present is an assessment of the neuroprotective state and has already been established as an established method. It has been reported.
- the mouse was euthanized, the eyeball was removed, and then the eyeball tissue was chemically fixed with formalin.
- the fixed eyeball was immersed in an OCT compound and frozen and solidified, and then thinned with a cryostud.
- the obtained sections were stained with hematoxylin and eosin (HE staining), and the structure of the retina was observed with an optical microscope.
- FIG. 1 The results are shown in FIG.
- (a) is the retina of a normal mouse
- (b) to (h) are the retina of a retinal degeneration mouse
- the instilled solution is as follows: (b) Calpain inhibitor I nanoparticle water (C) Calpain inhibitor I suspension; (d) No instillation (control); (e) Base alone (control) (f) Calpeptin nanoparticle aqueous dispersion; (g) Calpeptin suspension; (h ) No instillation (control).
- N indicates the optic nerve head.
- the eye drop group of the suspension (FIGS. 14 (c) and (g)) and the eye drop group of the suspension (FIGS. 14 (c) and (g)) were compared, the cell density was overwhelming. It is clear that the eye drop group of the nanoparticle aqueous dispersion is higher, has no vacuoles, and apoptosis is suppressed. Thus, it was demonstrated that the eye drop of the present invention has a therapeutic effect on retinal degenerative diseases.
- the present invention can be the following embodiments: [1] It includes a step of freeze-drying a sample obtained by freezing a mixture of the first solution and the second solution, and a step of dispersing the freeze-dried sample in water, wherein the mixture comprises an active ingredient and an ointment base
- the ointment base is lanolin, petrolatum, beeswax, phenol / zinc flower liniment, cocoa butter, witepsol, glycerogelatin, liquid paraffin, hard fat, macrogol, hydrocarbon gel ointment base, or a derivative thereof.
- One method. [3] The method of 1 or 2, wherein the active ingredient is a poorly water-soluble or water-insoluble drug.
- the method of 1 to 3 further comprising the step of confirming that no crystal growth of nanoparticles containing the active ingredient has occurred in the dispersion obtained by the step of dispersing.
- the method of 4, wherein the step of confirming is a step of filtering the obtained dispersion.
- composition according to 15 or 16 for delivering an active ingredient of a drug from an applied site to a deep tissue site.
- the 17 compositions for treating a disease in a deep eye part.
- the disease is selected from the group consisting of retinal dystrophy, retinitis pigmentosa, glaucoma, age-related macular degeneration, diabetic retinopathy, and retinal and neurodegenerative diseases that occur secondary to various diseases.
- Composition [20] The composition of 15 to 19, which is used as a sustained release preparation.
- the ointment base is lanolin, petrolatum, beeswax, phenol / zinc flower liniment, cocoa butter, witepsol, glycerogelatin, liquid paraffin, hard fat, macrogol, hydrocarbon gel ointment base, or a derivative thereof. There are 21 methods.
- the present invention can provide a technology for suppressing the crystal growth of drug nanoparticles in water, it will play an extremely important role in the field of pharmaceutical preparations using drug nanoparticles, which will be intensified in the future. .
Abstract
Description
本発明は、ナノ粒子水分散液を製造する方法を提供する。特に、本発明は、難水溶性または非水溶性の薬剤(またはその有効成分)を首尾よく分散した薬剤ナノ粒子水分散液を製造する方法を提供する。本発明の製造方法は、軟膏基剤が目的の有効成分(例えば、難水溶性または非水溶性の薬剤)とともに含まれた、有機溶媒と水との混合液を凍結し、次いで凍結サンプルを凍結乾燥し、さらに凍結乾燥サンプルを水に再分散させることによって行われる。すなわち、本発明の製造方法は、ナノ粒子水分散液を製造するために、溶媒に有機溶媒を用いた第一溶液と溶媒に水を用いた第二溶液との混合液を凍結したサンプルを凍結乾燥する工程、および、凍結乾燥したサンプルを水に分散させる工程、を包含し、上記混合液は有効成分および軟膏基剤を含有している。
本発明はまた、上述した方法によって製造されたナノ粒子水分散液を提供する。本発明のナノ粒子水分散液は、軟膏基剤が目的の有効成分(難水溶性または非水溶性の薬剤)とともに含まれた、ナノメートルのサイズの粒径を有する粒子が水に分散している。本発明のナノ粒子水分散液に分散しているナノ粒子に含まれる軟膏基剤は、上述したものであれば特に限定されない。
本発明は、ナノ粒子水分散液を含有している組成物を提供する。本発明の組成物は、上記ナノ粒子水分散液の性質に起因して種々の用途に利用され得る。
軟膏(または軟膏剤)は、ワセリン等の基剤の中に有効成分を分散させた、半固形の外用剤であり、患部に塗布されることによって皮膚疾患等の治療に用いられる。軟膏に用いられる基剤(軟膏基剤ともいう。)としては、ワセリン等の疎水性基剤(油脂性基剤)、親水性基剤(乳剤性基剤、水溶性基剤、懸濁性基剤)、疎水性基剤、糊膏(リニメント)、泥膏(パスタ)、硬膏、ローション、スプレー等が挙げられ、軟膏の使用形態は、基本的にクリーム、ゲル、ローション等の塗布型である。軟膏は、軟膏基剤の性質によって、皮膚に付着することによって有効成分を長く皮膚に留めることができ、皮膚外用剤や、口腔用軟膏、眼軟膏等に使用されている。
本発明に用いられる薬剤は、主に難水溶性薬剤であり、ナノサイズ化されている。上述したように、本発明の組成物は製剤がナノサイズで提供され、組織や細胞への浸透性が向上する。これに起因して、本発明の組成物は、薬剤の最小投与による最大効果を見込めるDDS(ドラッグデリバリーシステム)に極めて有効な剤形であり、優れた薬理効果や患者へのコンプライアンスの向上が期待される。すなわち、本発明の組成物は、適用した部位(組織表面)から組織深遠部へ薬剤の有効成分を送達することが必要な局面に利用される。上述した点眼剤として用いられた場合、本発明の組成物は、眼深遠部(網膜等)における疾患を処置するために用いられる。このように、本発明の組成物は、新たな剤形を提供するので、多くの医療分野での活用や抜群の経済効果を期待し得る。
本発明はさらに、ナノ結晶の成長を抑制する方法を提供する。薬剤ナノ結晶の結晶成長を抑制することは、今後開発が熾烈になるナノ結晶製剤の製剤化において極めて重要な課題である。本発明は、軟膏基剤を用いて、薬剤ナノ結晶の表面をコーティングすることによって結晶成長を抑制する方法を提供する。本発明が適用された薬剤ナノ結晶は、コーティングされた軟膏製剤の特徴点(徐放性、滞留性)を保持した、新たな製剤形態にて提供される。
デキサメタゾン(30mg)、軟膏基剤であるラノリン(60mg)、およびポリオキシエチレン(200)ポリオキシプロピレングリコール(70)(60mg)を10mLのt-ブチルアルコールに溶解して溶液A(第一溶液)を調製した。また、ポリビニルピロリドン(20mg)、ヒドロキシプロピルメチルセルロース(3mg)、およびポリソルベート80(5ul)を10mLの精製水に溶解して溶液B(第二溶液)を調製した。
得られたデキサメタゾンのナノ粒子水分散液におけるデキサメタゾンのナノ結晶の電子顕微鏡像を図1に示す。コントロールとして、従来技術の手順に従って超音波懸濁によってデキサメタゾンの水分散液を作製し、水分散液中のデキサメタゾンのナノ結晶を電子顕微鏡にて観察した(図2)。
上述したデキサメタゾンのナノ粒子水分散液(A)、従来技術によって作製したデキサメタゾンの水分散液(B)、およびフルオロメトロンのナノ粒子水分散液(C)を、別々のバイアルに入れて観察した(図4)。正立したバイアル(左図)を倒し、背後に配置した文字をバイアルの側方より観察した。AおよびCでは、バイアル中の分散液を介して背後の文字を確認することができたことから、分散した粒子の粒径が小さいこと、および粒子が均一に分散したために光散乱が低下していることがわかった。しかし、Bでは、バイアル中の分散液を介して背後の文字を確認することができなかったことから、分散した粒子の粒径が大きいこと、および粒子の分散が不均一なために強く光散乱が生じたことがわかった。
本発明のナノ粒子水分散液において結晶の成長が抑制されているか否かを検証するために、ラノリンを使用したフルオロメトロン水分散液A(すなわち本発明のナノ粒子分散液)とラノリンを使用しないフルオロメトロン水分散液Bを作製した。分散液AおよびBを60℃のオイルバスに48時間静置するという過酷試験を行い、過酷試験の前と後との間でどの程度の結晶成長が生じているかを、電子顕微鏡観察、ならびに動的光散乱法による粒度分布測定およびゼータ電位測定によって調べた。その結果、分散液Aでは、過酷試験の前後で粒子径、粒度分布およびゼータ電位にほとんど変化が見られなかったが、分散液Bでは、過酷試験の後にナノ粒子の結晶成長、粒度分布の広がりおよびゼータ電位の大幅な低下による粒子分散の不安定化が見られた。電子顕微鏡像を図5に、粒度分布およびゼータ電位を図6に示す。このように、本発明のナノ粒子分散液におけるナノ粒子の結晶成長抑制効果が実証された。
軟膏基剤としてラノリンを用いたナノ粒子水分散液の作製手順と同様の手順に従って、軟膏基剤として(a)ワセリン、(b)蜜蝋、(c)ウイテプゾール、または(d)カカオ脂を用いた、フルオロメトロンのナノ粒子水分散液を得た。得られたナノ粒子水分散液におけるナノ結晶の電子顕微鏡像を図8に示す。図1と同様に、200nm程度のナノ結晶群が観察され、本発明の方法に従えば良好なナノ粒子を得ることができることがわかった。さらに、得られたナノ粒子水分散液におけるナノ結晶の、動的光散乱法による粒度分布およびゼータ電位の結果を、表1に示す。分散した粒子の粒径が小さいこと、粒子が均一に分散したために光散乱が低下していること、およびゼータ電位測定によって表面がネガティブにチャージしたナノ粒子が安定的に分散していることがわかった。
ステロイド(デキサメタゾンまたはフルオロメトロン)のナノ粒子水分散液の作製手順と同様の手順に従って、薬剤としてステロイド以外のものを用いてナノ粒子水分散液を得た。得られたナノ粒子水分散液におけるナノ結晶の電子顕微鏡像を図10に示す。図中、(a)~(d)はそれぞれ、薬剤としてカルパインインヒビターI、カルペプチン、シクロスポリンA、7-エチル-10-ヒドロキシカンプトテシンを用いた場合のナノ結晶像である。図1と同様に、50~400nm程度のナノ結晶群が観察され、本発明の方法に従えば良好なナノ粒子を得ることができることがわかった。さらに、種々の薬剤において得られたナノ粒子水分散液におけるナノ結晶の、動的光散乱法による粒度分布およびゼータ電位の結果を、表2に示す。表中のALLNおよびSN38はそれぞれカルパインインヒビターIおよび7-エチル-10-ヒドロキシカンプトテシンを示す。分散した粒子の粒径が小さいこと、粒子が均一に分散したために光散乱が低下していること、およびゼータ電位測定によって表面がネガティブにチャージしたナノ粒子が安定的に分散していることがわかった。また、本発明を用いれば、低分子化合物だけでなくペプチド(カルペプチン)または油滴状(クロフィブラート)の薬剤をナノ粒子化することができることがわかった。
本発明のナノ粒子水分散液を用いることによって化合物(ナノ結晶)が目的の組織へ長期的に滞留することができるか否かを検証するために、ラノリンを使用した水分散液E(すなわち本発明のナノ粒子分散液)とラノリンを使用しない水分散液Fを作製した。蛍光色素としてフルオレセイン(CAS 2321-07-5, Aldrich社)を使用した各水分散液の作製手順は以下のとおりである:
(1)ラノリン(60mg)、ユニルーブ(60mg)および蛍光色素フルオレセイン(1mg)をt-ブチルアルコール(10mL)に溶解させた溶液A(第一溶液)、ならびにPVP(20mg)、Tween 80(5μL)およびヒドロキシプロピルメチルセルロース(HPMC)(3mg)を溶解させた溶液B(第二溶液)を用いて、上述した手順に従って、ラノリンを含むフルオレセイン水分散液Eを作製した;
(2)ユニルーブ(60mg)および蛍光色素フルオレセイン(1mg)をt-ブチルアルコール(10mL)に溶解させた溶液A(第一溶液)、ならびにPVP(20mg)、Tween 80(5μL)およびヒドロキシプロピルメチルセルロース(HPMC)(3mg)を溶解させた溶液B(第二溶液)を用いて、上述した手順に従って、ラノリンを含まないフルオレセイン水分散液Fを作製した。
本発明のナノ粒子水分散液を用いることによって化合物(ナノ結晶)が眼球だけでなく皮膚へも長期的に滞留することができるか否かを、上記水分散液E(すなわち本発明のナノ粒子分散液)と上記水分散液Fを用いて検証した。
本発明のナノ粒子水分散液を用いて点眼剤を作製し、眼深部疾患である網膜ぶどう膜炎に対する抗炎症効果を検証した。網膜ぶどう膜炎の抗炎症効果の評価にヒト内因性ぶどう膜炎の動物モデルである実験的自己免疫性網膜ぶどう膜炎(EAU:experimental autoimmune uveoretinitis)を用いた。
本発明のナノ粒子水分散液を用いて点眼剤を作製し、ウサギに点眼した薬物の眼内移行性を評価した。フルオロメトロンを含有しない点眼基剤のみのコントロール液を施した系(1眼)、市販の点眼剤(フルオロメトロン濃度0.1重量%)を施した系(1眼)、本発明の点眼剤(フルオロメトロン濃度0.1重量%)を施した系(1眼)、点眼を施さない系(1眼)について点眼を行った。
本発明のナノ粒子水分散液を用いて点眼剤を作製し、網膜変性疾患の治療効果を検証した。薬剤にカルパインインヒビター(カルパインインヒビターIおよびカルペプチン)を用い、先天的に網膜変性を示すマウス(C3H/HeNCrlCrlj)に対して生後5日目から1日2回点眼を開始した。コントロールには、粒子サイズが粗大な(すなわちナノ粒子でない)懸濁型のカルパインインヒビターの点眼液、およびカルパインインヒビターを含まない基剤のみの点眼液を用いた。C3H/HeNCrlCrljマウスは、ホスホジエステラーゼ(PDE)6Bに遺伝子変異を有するマウスであり、生後すぐに網膜変性が始まり、生後2週目にはかなりの網膜色素上皮細胞および視細胞がアポトーシスに陥る、ヒトにおける網膜色素変性のモデル動物と考えられている。視細胞生存の評価の上で、視細胞の核が存在する外顆粒層の細胞密度や層構造の状態を評価することは、神経保護の状態を評価することになり、確立された方法として既に報告されている。
[1]第一溶液と第二溶液との混合液を凍結したサンプルを凍結乾燥する工程、および、凍結乾燥したサンプルを水に分散させる工程、を包含し、上記混合液は有効成分および軟膏基剤を含有し、第一溶液の溶媒は有機溶媒であり、第二溶液の溶媒は水である、有効成分のナノ粒子水分散液を製造する方法。
[2]前記軟膏基剤が、ラノリン、ワセリン、蜜蝋、フェノール・亜鉛華リニメント、カカオ脂、ウイテプゾール、グリセロゼラチン、流動パラフィン、ハードファット、マクロゴール、ヒドロカーボンゲル軟膏基剤、またはこれらの誘導体である、1の方法。
[3]前記有効成分が難水溶性または非水溶性の薬剤である、1または2の方法。
[4]前記分散させる工程によって得られた分散液において、有効成分を含むナノ粒子の結晶成長が生じていないことを確認する工程をさらに包含する、1~3の方法。
[5]前記確認する工程が、前記得られた分散液を濾過する工程である、4の方法。
[6]前記分散させる工程によって得られた分散液において、沈殿が生じていないことを確認する工程をさらに包含する、1~5の方法。
[7]前記確認する工程を、前記分散させる工程の1週間以上後に行う、4~6の方法。
[8]前記分散させる工程によって得られた分散液から、有効成分を含むナノ粒子の結晶成長が生じていない分散液を選択する工程をさらに包含する、4~7の方法。
[9]前記分散させる工程によって得られた分散液から、沈殿が生じていない分散液を選択する工程をさらに包含する、6または7の方法。
[10]第一溶液と第二溶液との混合液を凍結する工程をさらに包含する、1~9の方法。
[11]前記混合液を凍結する工程が液体窒素環境下にて行われる、10の方法。
[12]第一溶液と第二溶液との混合液を作製する工程をさらに包含する、1~11の方法。
[13]有効成分および軟膏基剤を有機溶媒に溶解して第一溶液を作製する工程、および/または、分散剤を水に溶解して第二溶液を作製する工程をさらに包含する、1~12の方法。
[14]1~13の方法によって製造された、ナノ粒子水分散液。
[15]14のナノ粒子水分散液を含有している、組成物。
[16]点眼剤または皮膚外用剤である、15の組成物。
[17]適用した部位から組織深遠部へ薬剤の有効成分を送達するための、15または16の組成物。
[18]眼深遠部における疾患を処置するための、17の組成物。
[19]前記疾患が、網膜ジストロフィー、網膜色素変性、緑内障、加齢黄斑変性、糖尿病網膜症、ならびに種々の疾患に続発して発生する網膜変性および神経変性疾患からなる群より選択される、18の組成物。
[20]徐放性製剤として用いられる、15~19の組成物。
[21]第一溶液と第二溶液との混合液を凍結したサンプルを凍結乾燥する工程を包含し、上記混合液は有効成分および軟膏基剤を含有し、第一溶液の溶媒は有機溶媒であり、第二溶液の溶媒は水である、有効成分のナノ結晶の成長を抑制する方法。
[22]前記軟膏基剤が、ラノリン、ワセリン、蜜蝋、フェノール・亜鉛華リニメント、カカオ脂、ウイテプゾール、グリセロゼラチン、流動パラフィン、ハードファット、マクロゴール、ヒドロカーボンゲル軟膏基剤、またはこれらの誘導体である、21の方法。
[23]前記有効成分が難水溶性または非水溶性の薬剤である、21または22の方法。
[24]前記凍結乾燥する工程によって得られたサンプルを水に分散させる工程をさらに包含する、21~23の方法。
[25]前記分散させる工程によって得られた分散液において、有効成分を含むナノ粒子の結晶成長が生じていないことを確認する工程をさらに包含する、24の方法。
[26]前記確認する工程が、前記得られた分散液を濾過する工程である、25の方法。
[27]前記分散させる工程によって得られた分散液において、沈殿が生じていないことを確認する工程をさらに包含する、22~24の方法。
[28]前記確認する工程を、前記分散させる工程の1週間以上後に行う、25~27の方法。
[29]前記分散させる工程によって得られた分散液から、有効成分を含むナノ粒子の結晶成長が生じていない分散液を選択する工程をさらに包含する、25~28の方法。
[30]第一溶液と第二溶液との混合液を凍結する工程をさらに包含する、21~29の方法。
[31]前記混合液を凍結する工程が液体窒素環境下にて行われる、30の方法。
[32]第一溶液と第二溶液との混合液を作製する工程をさらに包含する、21~31の方法。
[33]有効成分および軟膏基剤を有機溶媒に溶解して第一溶液を作製する工程、および/または、分散剤を水に溶解して第二溶液を作製する工程をさらに包含する、21~32の方法。
Claims (12)
- 有効成分および軟膏基剤を含有する、有機溶媒を溶媒とする第一溶液と水を溶媒とする第二溶液との混合液を凍結したサンプルを凍結乾燥する工程、および
凍結乾燥したサンプルを水に分散させる工程
を包含する、有効成分を含むナノ粒子を分散した水分散液を製造する方法。 - 前記軟膏基剤が、ラノリン、ワセリン、蜜蝋、フェノール・亜鉛華リニメント、カカオ脂、ウイテプゾール、グリセロゼラチン、流動パラフィン、ハードファット、マクロゴール、ヒドロカーボンゲル軟膏基剤、またはこれらの誘導体である、請求項1に記載の方法。
- 前記分散させる工程によって得られた分散液において、有効成分を含むナノ粒子の結晶成長が生じていないことを確認する工程をさらに包含する、請求項1または2に記載の方法。
- 前記確認する工程が、前記得られた分散液を濾過する工程である、請求項3に記載の方法。
- 前記分散させる工程によって得られた分散液から、有効成分を含むナノ粒子の結晶成長が生じていない分散液を選択する工程をさらに包含する、請求項3または4に記載の方法。
- 請求項1~5のいずれか一項に記載の方法によって製造された、水分散液。
- 請求項6に記載の水分散液を含有している、組成物。
- 点眼剤または皮膚外用剤である、請求項7に記載の組成物。
- 適用した部位から組織深遠部へ薬剤の有効成分を送達するための、請求項7または8に記載の組成物。
- 眼深遠部における疾患を処置するための、請求項9に記載の組成物。
- 前記疾患が、網膜ジストロフィー、網膜色素変性、緑内障、加齢黄斑変性、糖尿病網膜症、ならびに種々の疾患に続発して発生する網膜変性および神経変性疾患からなる群より選択される、請求項10に記載の組成物。
- 徐放性製剤として用いられる、請求項7~11のいずれか1項に記載の組成物。
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US11147811B2 (en) | 2016-03-10 | 2021-10-19 | Sumitomo Dainippon Pharma Co., Ltd. | Composition comprising fine particle and process thereof |
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WO2017120207A1 (en) * | 2016-01-05 | 2017-07-13 | Nanotech Industrial Solutions, Inc. | Water based nanoparticle dispersion |
WO2018162271A1 (en) * | 2017-03-05 | 2018-09-13 | Rita Dobmeyer | Nanoparticles for the treatment of macular degeneration |
CN110759928B (zh) * | 2018-07-27 | 2022-04-01 | 四川大学 | 利用可逆分解法制备喜树碱类药物纳米晶体 |
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