External preparation comprising non-lamellar liquid crystal-forming lipid

Abstract

To provide an external preparation that is well retained on a living body surface and is capable of increasing drug permeability.SOLUTION: The present invention provides an external preparation comprising a non-lamellar liquid crystal-forming lipid and a drug.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an external preparation comprising a non-lamellar liquid crystal forming lipid. [Background technology]

[0002] Transdermal preparations, which are applied to the skin, have the advantages of easy administration and sustained release, and are increasingly being used for not only local administration but also systemic administration of various drugs. However, since the skin has the function of restricting the permeation of substances as a biological barrier, it is known that the drug absorption in transdermal preparations is basically low. Therefore, in order to promote the permeation of drugs through the skin, various percutaneous absorption enhancers are used in transdermal preparations, but the percutaneous absorption enhancement effect obtained is not necessarily sufficient.

[0003] Lyotropic liquid crystals such as liposomes have been reported to be useful as biomimetic drug delivery system (DDS) carriers since the concept of DDS was first proposed. In recent years, nonlamellar liquid crystals (NLLCs), a type of lyotropic liquid crystal, have been reported to have advantages over conventional DDS carriers, such as high drug content, ease of preparation, and high stability in polymeric drugs.

[0004] Various liquid crystal-forming compounds are used for various purposes in the fields of cosmetics and medicines. In recent years, lipid compounds capable of forming cubic liquid crystals that are highly stable even at low temperatures (below 6°C) have been developed, and the use of these liquid crystals in sustained-release preparations has also been reported (Patent Document 1). However, these lipid compounds have high viscosity and cannot be passed through thin injection needles (e.g., 30 gauge), making them difficult to use in injections. Therefore, lipid compounds with lower viscosity that stably form non-lamellar liquid crystals have been developed as bases for injections (Patent Document 2). Patent Document 3 discloses a skin topical preparation containing liquid crystals formed by this low-viscosity lipid compound, but its main dosage form is a lotion or emulsion, and its skin retention is not high. Patent Document 3 also does not describe the application of the topical preparation to tissues other than the skin. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2006 / 043705 [Patent Document 2] International Publication No. 2011 / 078383 [Patent Document 3] JP 2012-17318 A Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide an external preparation that is well retained on the surface of a living body and can increase drug permeability. [Means for solving the problem]

[0007] The inventors have conducted extensive research to solve the above problems, and as a result of producing preparations such as patches such as tapes, aerosols, and liquid crystal precursor preparations using non-lamellar liquid crystal forming lipids, they have found that good preparation retention and increased drug permeability can be achieved on the biological surface, and that such preparations are suitable for application not only to the skin but also to mucous membranes, thereby completing the present invention.

[0008] That is, the present invention includes the following. [1] An external preparation comprising a non-lamellar liquid crystal forming lipid and a drug. [2] The topical preparation according to the above item [1], which is an amphiphilic compound represented by the following general formula (I) or a salt thereof: [ka] (In the formula, X and Y each represent a hydrogen atom or together represent an oxygen atom; n represents an integer of 0 to 2; m represents 1 or 2; TIFF2025029135000002.tif696 represents a single bond or a double bond, and R represents a hydrophilic group having two or more hydroxyl groups. [3] The topical preparation according to the above item [2], wherein R in the formula represents a hydrophilic group from which one hydroxyl group has been removed from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol. [4] The topical preparation according to any one of the above [1] to [3], wherein the non-lamellar liquid crystal forming lipid is mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol or mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol. [5] The topical preparation described in [1] above, wherein the non-lamellar liquid crystal forming lipid is glyceryl monooleate or phytantriol. [6] The external preparation according to any one of the above [1] to [5], which is formulated in the form of a patch. [7] The external preparation described in [6] above, wherein the patch is a tape. [8] The topical preparation according to [6] or [7] above, containing 70 w / w% or more of an adhesive. [9] The topical preparation according to any one of the above [1] to [5], which is formulated in an aerosol dosage form.

[10] The topical preparation according to any one of the above [1] to [9], wherein the non-lamellar liquid crystal forming lipid does not form liquid crystals in the topical preparation.

[11] The external preparation according to any one of the above [1] to

[10] , for application to mucous membranes.

[12] The topical preparation according to any one of the above [1] to

[11] , further comprising a water-soluble polymer and / or an oil component.

[13] The topical preparation described in

[12] above, wherein the water-soluble polymer is hydroxypropyl cellulose.

[14] The topical preparation according to any one of the above [1] to

[13] , further comprising ethanol.

[15] The topical preparation according to any one of the above [1] to [5], which comprises fine particles containing the non-lamellar liquid crystal forming lipid and the drug.

[16] The external preparation according to any one of the above [1] to

[15] , for drug delivery into the brain. This specification includes the disclosure of Japanese Patent Application No. 2018-168365, which is the priority basis of this application. Effect of the Invention

[0009] According to the present invention, it is possible to provide an external preparation that is well retained on the surface of a living body and can increase the skin permeability of a drug, and also to provide an external preparation that can bring about high mucosal permeability of a drug when applied to a mucosa. [Brief description of the drawings]

[0010] [Figure 1] Figure 1 shows photographs of the polarizing microscope images: A: Formulation No. 1, B: Formulation No. 2, C: Formulation No. 3, D: Formulation No. 4, E: Formulation No. 5, F: Formulation No. 19, G: Formulation No. 25, H: Formulation No. 26. [Diagram 2] 2 shows the results of small angle X-ray diffraction: A: Formulation No. 1, B: Formulation No. 2, C: Formulation No. 3, D: Formulation No. 13, E: Formulation No. 25, F: Formulation No. 26, G: Formulation No. 27. [Diagram 3] 3 shows the results of small angle X-ray diffraction: A: Formulation No. 37, B: Formulation No. 45, C: Formulation No. 46, D: Formulation No. 47, E: Formulation No. 48, F: Formulation No. 49. [Figure 4] FIG. 4 is a schematic diagram of the structure of a vertical diffusion cell. [Diagram 5] FIG. 5 shows the skin permeation behavior of FL-Na from a liquid crystal precursor formulation. [Figure 6] 6 shows the results of small angle X-ray diffraction: A: Formulation No. 52 (water added), B: Formulation No. 53 (water added), C: Formulation No. 56 (water added), D: Formulation No. 57 (water added), E: Formulation No. 56 (water not added). [Figure 7] 7 shows the skin permeation behavior of FL-Na from a spray formulation. Each value is the mean ± standard error (SE). [Figure 8] 8 is a schematic diagram showing the spreading of an adhesive layer on a liner, with the arrows indicating the spreading direction of the adhesive layer. [Figure 9] Figure 9 is a photograph showing a phase image of the adhesive layer surface of a tape formulation. A: Formulation No. 73, observation field 1 μm x 1 μm. B: Formulation No. 63, observation field 1 μm x 1 μm. C: Formulation No. 73, observation field 0.5 μm x 0.5 μm. D: Formulation No. 63, observation field 0.5 μm x 0.5 μm. [Figure 10] FIG. 10 is a schematic diagram of the structure of a horizontal diffusion cell. [Figure 11] FIG. 11 shows the release profile of FL-Na from the tape preparation. [Figure 12] FIG. 12 shows the skin permeation behavior of FL-Na from a tape preparation. [Figure 13] 13 shows the results of small angle X-ray diffraction: A: Preparation No. 75, B: Preparation No. 76, C: Preparation No. 77. [Figure 14] 14 is a diagram showing the release profile of tranilast from each formulation 8 hours after application. Each value represents the mean ± standard error (SE). The open squares represent formulation No. 75, the filled squares represent formulation No. 76, and the open triangles represent formulation No. 78. [Figure 15] 15 is a graph showing the change in plasma tranilast concentration over 8 hours after intranasal administration. Each value represents the mean ± standard error (SE). Open squares represent formulation No. 75, filled squares represent formulation No. 76, open circles represent formulation No. 77, and open triangles represent formulation No. 78. [Figure 16] 16 is a graph showing the change in the tranilast concentration in the brain 8 hours after intranasal administration. Each value represents the mean ± standard error (SE). The open squares represent formulation No. 75, the filled squares represent formulation No. 76, the open circles represent formulation No. 77, and the open triangles represent formulation No. 78. [Figure 17] Figure 17 shows the tranilast concentrations in different regions of the brain 2, 4 and 8 hours after intranasal administration: A: midbrain, B: cortex, C: cerebellum, D: hippocampus. [Figure 18]Figure 18 shows the tranilast concentration in different regions of the brain 2, 4 and 8 hours after intranasal administration: A: spinal cord, B: olfactory bulb. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] The present invention will be described in detail below.

[0012] The present invention relates to an external preparation comprising a non-lamellar liquid crystal forming lipid and a drug. In the present invention, the external preparation refers to a medicine to be applied to a biological surface (such as the skin or mucosa) for the purpose of administering a drug. Such an external preparation may be a pharmaceutical composition. It is preferable that the external preparation according to the present invention is adhesive to a biological surface and is stably retained on the biological surface. When the external preparation according to the present invention is applied to a biological surface, the drug is released from the liquid crystal formed by the non-lamellar liquid crystal forming lipid in the external preparation, and the drug efficiently permeates the skin, mucosa, etc. and is absorbed (administered) into the body. The external preparation according to the present invention can significantly promote the permeation of the drug through the skin, mucosa, etc.

[0013] 1. Non-lamellar liquid crystal forming lipids In the present invention, lipids capable of forming non-lamellar liquid crystals (non-lamellar liquid crystal forming lipids) can be used as liquid crystal forming lipids. The non-lamellar liquid crystal forming lipids used in the present invention are preferably low molecular weight amphipathic compounds. Here, "low molecular weight" means having a molecular weight of about 20 to 10,000. The molecular weight of the non-lamellar liquid crystal forming lipids used in the present invention is preferably 50 to 5,000, more preferably 100 to 2,500, and even more preferably 200 to 1,000.

[0014] In one embodiment, an amphiphilic compound represented by the following general formula (I) or a salt thereof can be used as the non-lamellar liquid crystal forming lipid.

[0015] [ka] In the general formula (I), X and Y each represent a hydrogen atom or together represent an oxygen atom. In the general formula (I), n represents an integer of 0 to 2 (preferably 1 or 2), and m represents 1 or 2. In the amphiphilic compound represented by the general formula (I), the combination of n and m may be any of n=0, m=1; n=0, m=2; n=1, m=1; n=1, m=2; n=2, m=1; or n=2, m=2.

[0016] In the formula: TIFF2025029135000004.tif696 represents a single bond or a double bond.

[0017] R in the general formula (I) represents a hydrophilic group having two or more hydroxyl groups, and is not limited to the following, but examples thereof include hydrophilic groups obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol. R in the general formula (I) is more preferably a hydrophilic group obtained by removing one hydroxyl group (OH) from glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid, or xylose, and is particularly preferably a hydrophilic group obtained by removing one hydroxyl group (OH) from glycerol. The hydrophilic group obtained by removing one hydroxyl group (OH) from glyceric acid may be a group obtained by removing an OH (hydroxyl group) contained in a carboxyl group of glyceric acid.

[0018] In the present invention, the notation in general formula (I): TIFF2025029135000005.tif898 means that the amphiphilic compound is a geometric isomer of E form (cis form) or Z form (trans form), or a mixture thereof. The meaning of this notation is the same in the general formulas (II) and (III) described below.

[0019] An example of the amphiphilic compound represented by general formula (I) is the amphiphilic compound represented by the following general formula (II).

[0020] [ka] In general formula (II), X and Y each represent a hydrogen atom or together represent an oxygen atom; n represents an integer of 0 to 2 (0, 1 or 2); and m represents 1 or 2.

[0021] R in the general formula (II) represents a hydrophilic group obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol. A preferred example of R is a hydrophilic group obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid, and xylose, and more preferably a hydrophilic group obtained by removing one hydroxyl group (OH) from glycerol. The hydrophilic group obtained by removing one hydroxyl group (OH) from glyceric acid may be a group obtained by removing an OH (hydroxyl group) contained in a carboxyl group of glyceric acid.

[0022] Another example of the amphiphilic compound represented by general formula (I) is the amphiphilic compound represented by the following general formula (III).

[0023] [ka] In formula (III), X and Y each represent a hydrogen atom or together represent an oxygen atom; n represents an integer of 0 to 2 (preferably 1 or 2); and m represents 1 or 2.

[0024] R in the general formula (III) represents a hydrophilic group having two or more hydroxyl groups, and is not limited to the following, but examples thereof include hydrophilic groups obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol. A preferred example of R is a hydrophilic group obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid, and xylose, and more preferably a hydrophilic group obtained by removing one hydroxyl group (OH) from glycerol. The hydrophilic group obtained by removing one hydroxyl group (OH) from glyceric acid may be a group obtained by removing an OH (hydroxyl group) contained in a carboxyl group of glyceric acid.

[0025] Yet another example of the amphiphilic compound represented by general formula (I) is the amphiphilic compound represented by the following general formula (IV).

[0026] [ka] In formula (IV), X and Y each represent a hydrogen atom or together represent an oxygen atom; n represents an integer of 0 to 2 (preferably 1 or 2); and m represents 1 or 2.

[0027] R in the general formula (IV) represents a hydrophilic group having two or more hydroxyl groups, and is not limited to the following, for example, a hydrophilic group obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol. A preferred example of R is a hydrophilic group obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid, and xylose, and more preferably a hydrophilic group obtained by removing one hydroxyl group (OH) from glycerol. The hydrophilic group obtained by removing one hydroxyl group (OH) from glyceric acid may be a group obtained by removing an OH (hydroxyl group) contained in a carboxyl group of glyceric acid.

[0028] Preferred examples of the amphiphilic compound represented by the general formula (I) include Mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol, Mono-O-(5,9,13-trimethyltetradecanoyl)glycerol, Mono-O-(5,9,13-trimethyltetradeca-4,8,12-trienoyl)glycerol, Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol, Mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol, and Mono-O-(5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoyl)glycerol These include, but are not limited to:

[0029] More preferred examples include mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol and mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol.

[0030] The amphiphilic compound represented by the general formula (I) used in the present invention exhibits high stability under a wide range of environmental conditions. For example, the amphiphilic compound represented by the general formula (I) is characterized by having an isoprenoid chain as a hydrophobic group, and unlike an amphiphilic compound having a linear aliphatic chain such as oleic acid as a hydrophobic group, it has high resistance to hydrolysis and relatively high oxidation stability. The amphiphilic compound represented by the general formula (I) also has a wide temperature range in which it can form liquid crystals, a low Krafft temperature, and can stably form liquid crystals even at low temperatures (6°C or lower, preferably 0°C or lower).

[0031] Furthermore, the amphiphilic compound represented by general formula (I) used in the present invention itself exhibits low viscosity. Specifically, the amphiphilic compound represented by general formula (I) itself has a viscosity of preferably 15.0 Pa·s or less, more preferably 11.0 Pa·s or less, and even more preferably 6.0 Pa·s or less, measured at 25°C. This viscosity can be measured, for example, at a temperature of 25°C using a viscosity / viscoelasticity measuring device (Gemini II, Malvern Instruments).

[0032] The topical agent according to the present invention may contain a salt of an amphiphilic compound represented by general formula (I). The salt of an amphiphilic compound represented by general formula (I) according to the present invention may be any salt, including salts of alkali metals or alkaline earth metals such as sodium, potassium, calcium, magnesium, etc., and sodium salts and potassium salts are preferred. The salt of an amphiphilic compound represented by general formula (I) according to the present invention may be a pharma- ceutical acceptable salt or a cosmetic acceptable salt.

[0033] The present invention is not limited to topical preparations using an amphiphilic compound represented by general formula (I) or a salt thereof, and any other non-lamellar liquid crystal forming lipid may also be used.

[0034] The non-lamellar liquid crystal forming lipid used in the present invention may be, for example, a glycerin fatty acid monoester. The fatty acid constituting the glycerin fatty acid monoester is preferably a saturated or unsaturated fatty acid having 8 to 24 carbon atoms. Other examples of glycerin fatty acid monoester include, but are not limited to, glyceryl monooleate (GMO; also known as monoolein), glyceryl monoisostearate, and glyceryl monoelaidylate. Alternatively, a glycerin monoalkyl ether may be used as the non-lamellar liquid crystal forming lipid. Specific examples of the glycerin monoalkyl ether include, but are not limited to, glycerin monooleyl ether (also known as oleyl glyceryl), glycerin monoisostearyl ether (also known as isostearyl glyceryl), and glycerin monoelaidyl ether. Furthermore, phytantriol (PHY) and the like can also be used as another non-lamellar liquid crystal forming lipid.

[0035] The non-lamellar liquid crystal forming lipids used in the present invention include combinations of two or more lipids that form non-lamellar liquid crystals when mixed together, and lipids that form non-lamellar liquid crystals when combined with components such as oils. Such non-lamellar liquid crystal forming lipids are known to those skilled in the art.

[0036] The liquid crystal structure formed may be changed by adding one or more specific components such as oil to the non-lamellar liquid crystal forming lipid used in the present invention. Even if an external preparation contains a non-lamellar liquid crystal forming lipid and such a component and the liquid crystal structure changes compared to when the component is not contained, as long as the external preparation forms non-lamellar liquid crystals even after the change in the liquid crystal structure, the external preparation and the non-lamellar liquid crystal forming lipid used therein are included in the scope of "external preparation" and "non-lamellar liquid crystal forming lipid" in the present invention, respectively.

[0037] The non-lamellar liquid crystal forming lipid used in the present invention can form non-lamellar liquid crystals in an aqueous medium (aqueous phase). In the present invention, an aqueous medium containing a non-lamellar liquid crystal forming lipid may be referred to as a "non-lamellar liquid crystal forming lipid / aqueous system" or an "amphiphilic compound / aqueous system".

[0038] The non-lamellar liquid crystal forming lipid contained in the topical preparation according to the present invention forms non-lamellar liquid crystals in the topical preparation, or forms non-lamellar liquid crystals on the surface of a living body due to the presence of surrounding water when applied to the surface of a living body, or forms non-lamellar liquid crystals on the surface of a living body due to the evaporation of volatile components (solvents such as ethanol, propellants, etc.) when applied to the surface of a living body. The non-lamellar liquid crystal formed by the non-lamellar liquid crystal forming lipid contained in the topical preparation according to the present invention is preferably a type II (water-in-oil) liquid crystal in which hydrophobic groups are oriented outward, and more preferably a cubic liquid crystal, a reverse hexagonal liquid crystal, or a mixture thereof, but is not limited thereto.

[0039] The liquid crystal structure formed by non-lamellar liquid crystal forming lipids can be analyzed by standard methods such as observation with a polarizing microscope or small angle X-ray scattering (SAXS) measurement.

[0040] For example, to confirm the formation of liquid crystals, the presence of various liquid crystal structures may be examined by small angle X-ray scattering (SAXS). Usually, a non-lamellar liquid crystal forming lipid / water sample of a given concentration is first placed in, for example, a quartz X-ray capillary tube, and the capillary is then sealed with an oxygen burner and subjected to SAXS measurement.

[0041] The formation of liquid crystals can be confirmed by checking whether the SAXS measurement results show the following scattering peak ratios (peak intervals) that are specific to each liquid crystal structure. Pn3m Cubic LCD Ratio: √2:√3:√4:√6:√8:√9:√10: ,,,, Ia3d Cubic LCD Ratio: √3:√4:√7:√8:√10:√11: ,,,, Im3m Cubic LCD Ratio: √2:√4:√6:√8:√10:√12:√14: ,,,, Fd3m cubic LCD ratio: √3:√8:√11:√12:√16:√19:√24:√27: ,,,, Ratio specific to inverted hexagonal liquid crystal: 1:√3:2: ,,,, According to a method well known to those skilled in the art, the space group and lattice constant can be easily determined by calculating peak values ​​from SAXS data and then calculating the ratio of their reciprocals.

[0042] The amphiphilic compound represented by general formula (I) used in the topical agent of the present invention can be synthesized by referring to the description in the Examples below or according to the synthesis method described in International Publication WO2014 / 178256. Alternatively, the amphiphilic compound represented by general formula (III) can be synthesized, for example, according to the synthesis method described in International Publication WO2011 / 078383. Furthermore, the amphiphilic compound represented by general formula (IV) can be synthesized, for example, according to the synthesis method described in International Publication WO2006 / 043705.

[0043] It is preferable to confirm that the synthesized compound is the desired compound by a standard method such as NMR measurement.

[0044] Various other non-lamellar liquid crystal lipids are also commercially available. Glyceryl monooleate (GMO) and phytantriol (PHY) are commercially available from Tokyo Chemical Industry Co., Ltd., Kao Corporation, Riken Vitamin Co., Ltd. (all Japan), etc. Oleyl glyceryl is commercially available, for example, under the trade name NIKKOL Selakis Alcohol V from Nikko Chemicals Co., Ltd. (Japan). Isostearyl glyceryl is commercially available, for example, under the trade name Penetol GE-IS from Kao Corporation (Japan).

[0045] The topical preparation according to the present invention contains an effective amount of a non-lamellar liquid crystal forming lipid. The amount of the non-lamellar liquid crystal forming lipid contained in the topical preparation according to the present invention is not limited to the following, but is usually 0.1 w / w% or more, 0.5 w / w% or more, for example, 3 w / w% or more, 5 w / w% or more, 0.5 to 99.8 w / w%, 1 to 99.5 w / w%, 5 to 99 w / w%, 10 to 99.8 w / w%, 40 to 99.8 w / w%, based on the total weight of the topical preparation. / w%, 60 w / w% or more, 70 w / w% or more, 60 to 99.8 w / w%, 65 to 99.8 w / w%, 68 to 99 w / w%, 70 to 99.5 w / w%, 70 to 90 w / w%, 10 to 30 w / w%, 10 to 25 w / w%, 13 to 15 w / w%, 3 to 20 w / w%, 3 to 15 w / w%, 3 to 10 w / w%, or 5 to 9 w / w%.

[0046] The "total weight of the topical preparation" referred to here refers to the total weight of a composition (preferably a mixed or dispersed composition) used as a topical preparation, which contains at least a non-lamellar liquid crystal forming lipid and a drug, and does not include the weight of other components such as a support on which the composition is placed and a container in which the composition is contained.

[0047] 2. Drugs The drug incorporated in the topical preparation according to the present invention is released onto the surface of the body by applying the topical preparation to the surface of the body, and is absorbed into the body through the skin, mucous membrane, etc. The drug may be any substance (active ingredient) to be administered to the body. However, the drug is not the non-lamellar liquid crystal forming lipid itself. The drug may be an organic compound or an inorganic compound. The drug may be a water-soluble drug or a fat-soluble (lipophilic, water-insoluble or poorly water-soluble) drug. The drug may be a physiologically active substance. The drug may be, for example, a protein, a peptide, an amino acid, a nucleic acid, etc., but is not limited thereto. The drug may be, for example, an anticancer drug, an immunosuppressant, an analgesic (e.g., a non-opioid analgesic, an opioid analgesic such as morphine), an anti-inflammatory agent, an antiallergic agent (tranilast, etc.), a steroid drug (triamcinolone acetonide, etc.), an anti-obesity drug, an antidiabetic drug, an antibiotic, an antifungal agent, an antiviral agent, a vasodilator, an anesthetic, a smoking cessation aid (nicotine, etc.), an antipsychotic drug, an antihypertensive drug, a cardiac stimulant, a β-blocker, an anti-anemia drug, an antihyperlipidemic drug, a bronchodilator, a dementia drug, a brain / central nervous system disease drug for Alzheimer's disease, Parkinson's disease, a cerebrovascular disorder, or a brain tumor, a chronic obstructive pulmonary disease (COPD) drug, a glaucoma drug, a cataract drug, an age-related macular degeneration drug, an overactive bladder drug, an attention deficit / hyperactivity disorder drug, a hormone drug, a vaccine, and the like, but are not limited to these.

[0048] 3. Dosage form and composition of topical preparations The topical preparation according to the present invention may be for systemic administration or for local administration. The topical preparation according to the present invention may be formulated in any dosage form. The topical preparation according to the present invention may have the dosage form of, but is not limited to, a patch, for example, a tape (also called a plaster), a cataplasm, etc.; a spray, for example, an aerosol, a pump spray (manual or power spray), etc.; an ointment, a cream, an oral preparation, a nasal preparation, a suppository, a vaginal preparation, etc. A spray refers to a pharmaceutical preparation in a dosage form in which a drug is ejected by applying pressure by hand, power, a propellant (gas), or any other means. The topical preparation according to the present invention may contain fine particles (e.g., microparticles or nanoparticles) containing a non-lamellar liquid crystal forming lipid and a drug. The topical preparation according to the present invention may be a dispersion, and may contain, for example, a dispersion of the fine particles (e.g., microparticles or nanoparticles). The present invention also provides a formulation in any of these dosage forms containing the topical preparation according to the present invention.

[0049] The topical preparation according to the present invention may be for application to a biological surface, preferably to the skin (skin surface). Alternatively, the topical preparation according to the present invention may be for application to a mucosa (mucosal surface). The topical preparation according to the present invention can promote the permeation of a drug through the skin or mucosa. The topical preparation according to the present invention is suitable not only for application to the skin but also for application to the mucosa.

[0050] The topical preparation according to the present invention may contain an aqueous medium as another component. The aqueous medium may be, but is not limited to, water such as sterilized water, purified water, distilled water, ion-exchanged water, and ultrapure water; electrolyte aqueous solutions such as physiological saline, sodium chloride aqueous solution, calcium chloride aqueous solution, magnesium chloride aqueous solution, sodium sulfate aqueous solution, potassium sulfate aqueous solution, sodium carbonate aqueous solution, and sodium acetate aqueous solution; and buffer solutions such as phosphate buffer and Tris-HCl buffer. The aqueous medium is preferably physiologically acceptable water or aqueous solution. The aqueous medium may contain, for example, a component of the topical preparation, such as a drug, by dissolving, dispersing, or suspending.

[0051] The topical preparation according to the present invention may contain water-soluble polymers as other components. Examples of water-soluble polymers include, but are not limited to, hydroxypropyl cellulose (HPC), hydroxyethyl cellulose, polyvinylpyrrolidone, carbopol, carrageenan, chitosan, chondroitin acid salts, xanthan gum, hyaluronate (such as sodium hyaluronate), alginate (such as sodium alginate), gelatin, dextran, etc. Hydroxypropyl cellulose (HPC) includes, for example, five grades of HPC available from Nippon Soda Co., Ltd. (Japan): HPC-SSL (molecular weight about 40,000, viscosity 2-2.9 mPa·s), HPC-SL (molecular weight about 100,000, viscosity 3-5.9 mPa·s), HPC-L (molecular weight about 140,000, viscosity 6-10 mPa·s), HPC-M (molecular weight about 620,000, viscosity 150-400 mPa·s), and HPC-H (molecular weight about 910,000, viscosity 1000-4000 mPa·s). In one embodiment, the hydroxypropyl cellulose may have a molecular weight of 1,000,000 or less, or 800,000 or less, for example, 10,000-700,000 or 10,000-80,000.

[0052] The amount of the drug contained in the external preparation according to the present invention is not limited to the following, but may be typically 0.0001 w / w% or more, for example, 0.0001-10 w / w%, 0.0005-5 w / w%, 0.0005-1 w / w%, 0.001-5 w / w%, 0.001-1 w / w%, 0.001-0.1 w / w%, 0.001-0.05 w / w%, 0.001-0.01 w / w%, 0.01-5 w / w%, 0.01-1 w / w%, 0.01-0.1 w / w%, 0.05-1 w / w%, or 0.1-0.5 w / w%, based on the total weight of the external preparation. The meaning of "total weight of the external preparation" is as described above. In the present invention, w / w% means weight / weight% and is used interchangeably with mass / mass%.

[0053] The topical agent according to the present invention may contain oil as another component. The oil may be, but is not limited to, hydrocarbon oil, ester oil, vegetable oil, animal oil, and other oils and fats, higher alcohols such as behenyl alcohol and stearyl alcohol, higher fatty acids such as stearic acid and palmitic acid, and fat-soluble vitamins. Specific examples of oil may include, but are not limited to, squalene, squalane, isopropyl myristate, octyldodecyl myristate, castor oil, olive oil, tocopherol, and tocopherol acetate.

[0054] The topical preparation of the present invention may contain a surfactant as another component. Examples of surfactants used in the present invention include nonionic surfactants such as block copolymers of hydrophilic ethylene oxide and hydrophobic propylene oxide (polyoxyethylene polyoxypropylene glycol), polyoxyethylene alkyl ethers, polyoxyethylene alkyl esters, and polyoxyethylene hydrogenated castor oil. As nonionic surfactants, those having a molecular weight of 1000 or more (more preferably 5000 or more) are more preferred. Examples of block copolymers of ethylene oxide and propylene oxide include polyoxyethylene (200) polyoxypropylene (70) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol, polyoxyethylene (160) polyoxypropylene (30) glycol, and polyoxyethylene (120) polyoxypropylene (40) glycol. These block copolymers of ethylene oxide and propylene oxide are available from Pluronic. (R) , poloxamer (R) , Unilube (R) , Pronon (R) Particularly preferred examples of surfactants include polyoxyethylene (200) polyoxypropylene (70) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol (also known as Pluronic). (R) F127; Unilube 70DP-950B, Poloxamer (R)407) and the like, but are not limited to these. In the present invention, the non-lamellar liquid crystal forming lipids used in the present invention are not included in the scope of surfactants. The topical preparation of the present invention may contain one or more of such surfactants.

[0055] The topical preparation of the present invention may contain a surfactant in an amount of, based on the total weight of the topical preparation, typically 0.001 w / w% or more, for example, 0.01 w / w% or more, preferably 0.05 w / w% or more, more preferably 0.1 w / w% or more, for example, 0.01 to 10 w / w%, 0.1 to 5 w / w%, 0.3 w / w% or more, 0.3 to 2 w / w%, 0.3 to 1.5 w / w%, 0.3 to 1 w / w%, or 0.55 to 0.9 w / w%.

[0056] The topical preparation according to the present invention may contain ethanol (which means absolute ethanol unless otherwise specified in the present invention) as another component. Note that ethanol (absolute ethanol) is not an aqueous medium.

[0057] The topical preparation according to the present invention may contain, as other components, pharma- ceutically acceptable water-soluble organic compounds such as propylene glycol, glycerin, ethylene glycol, and butylene glycol.

[0058] The topical preparation according to the present invention may contain other pharma- ceutically acceptable additives as other components, including, but not limited to, carriers, excipients, stabilizers, buffers, preservatives, colorants, flavorings, pH adjusters, dispersants, etc.

[0059] In the topical preparation according to the present invention, the non-lamellar liquid crystal forming lipid may form liquid crystal (particularly non-lamellar liquid crystal), but may not. When the non-lamellar liquid crystal forming lipid does not form liquid crystal in the topical preparation, the topical preparation is also called a liquid crystal precursor preparation. In the topical preparation according to the present invention that does not contain an aqueous medium or does not contain a sufficient amount of the aqueous medium, the non-lamellar liquid crystal forming lipid does not form liquid crystal, so the topical preparation is a liquid crystal precursor preparation.

[0060] The topical agent according to the present invention can be applied to a living body surface, preferably to the skin or mucosa, to administer the drug contained in the topical agent to the living body. The present invention also provides a drug delivery method or drug administration method, which includes applying the topical agent according to the present invention to a living body surface, preferably to the skin or mucosa. The subject to which the topical agent according to the present invention is applied is not particularly limited, but is typically an animal, and preferably a mammal or a bird, including primates such as humans, livestock, pet animals such as dogs, cats, and rabbits, and laboratory animals. Preferred dosage forms of topical preparations are described in more detail below.

[0061] 4. Patches The topical preparation according to the present invention may be formulated in the form of a patch. The present invention also provides a patch containing the topical preparation according to the present invention. In the present invention, a patch refers to a pharmaceutical preparation that is attached to the skin or mucosa and is intended for transdermal or transmucosal absorption of a drug. The patch may be of local or systemic action. The patch has an adhesive layer. The adhesive layer of the patch is preferably a composition containing an adhesive (more preferably a fat-soluble adhesive), and preferably contains a non-lamellar liquid crystal forming lipid and a drug in addition to the adhesive. Patches include, but are not limited to, tapes (also called plasters) and cataplasms.

[0062] The tape preparation is a preparation having an adhesive layer using a pressure-sensitive adhesive as a base. The adhesive layer is preferably a composition containing a pressure-sensitive adhesive, a non-lamellar liquid crystal forming lipid, and a drug. In one embodiment, the tape preparation comprises an adhesive layer containing a pressure-sensitive adhesive, a non-lamellar liquid crystal forming lipid, and a drug, and a support. In a preferred embodiment, the tape preparation is a so-called matrix-type preparation comprising an adhesive layer containing a pressure-sensitive adhesive, a non-lamellar liquid crystal forming lipid, and a drug, a liner (release sheet), and a support. The pressure-sensitive adhesive used in the tape preparation is preferably a fat-soluble polymer. Examples of pressure-sensitive adhesives include, but are not limited to, acrylic, urethane, rubber, or silicone adhesives. Examples of acrylic adhesives include, but are not limited to, DURO-TAK(R) (Henkel), e.g. DURO-TAK (R) 387-2516 is an example.

[0063] The support may be of any shape that can be used for a patch, but is preferably a sheet-like substrate. For example, a known support can be used as the support. The support may be any material suitable for a patch support, such as a film such as a polymer film, a cloth such as a nonwoven fabric or a woven fabric, or paper. The support may be composed of, for example, polyester, polyethylene (polyethylene terephthalate, polyethylene naphthalate, polybutylene terephthalate, etc.), polyolefin (polyethylene, polypropylene, etc.), cellulose derivatives such as cellulose ester, polyurethane, polyamide, etc. The thickness of the support is not limited to the following, but is generally 5 μm to 500 μm, preferably 10 to 300 μm, for example 10 to 200 μm, 10 to 100 μm, 25 to 100 μm, 50 to 300 μm, or 60 to 200 μm.

[0064] As the release sheet, any one usable for a patch can be used, for example, a known release film can be used. The release sheet may be composed of, for example, a polymer film such as polyester (polyethylene terephthalate, polyethylene naphthalate, polybutylene terephthalate, etc.), polyolefin (polyethylene, polypropylene, etc.), polyvinyl chloride, polyvinylidene chloride, etc., a cellulose derivative such as cellulose ester, or paper, etc., or a laminate film using a plurality of these materials, etc., having a layer of fluororesin, silicone resin, etc. on the contact surface with the adhesive layer. The thickness of the release sheet is not limited to the following, but is generally 5 to 500 μm, preferably 10 to 300 μm, for example 10 to 200 μm, 25 to 100 μm, 50 to 300 μm, or 60 to 200 μm.

[0065] The thickness of the adhesive layer in the patch is not limited to the following, but is generally 5 μm to 1 mm, preferably 5 to 500 μm, for example 5 to 200 μm, 10 to 100 μm, or 20 to 50 μm.

[0066] The non-lamellar liquid crystal forming lipids and drugs are as described above.

[0067] In one embodiment, the amount of non-lamellar liquid crystal forming lipid in the adhesive layer of the patch, for example, the tape, is preferably an amount capable of forming liquid crystal. The non-lamellar liquid crystal forming lipid is as described above. More specifically, the amount of non-lamellar liquid crystal forming lipid in the adhesive layer is, but is not limited to, usually 0.1 w / w% or more, preferably 5 w / w% or more, for example, 1 to 20 w / w%, 1 to 10 w / w%, 5 to 20 w / w%, 5 to 10 w / w%, 10 to 30 w / w%, 10 to 25 w / w%, or 13 to 15 w / w% based on the total weight of the adhesive layer.

[0068] The amount of drug in the adhesive layer is not limited to the following, but is usually 0.0001 w / w% or more, preferably 0.0005 to 5 w / w%, more preferably 0.001 to 5 w / w%, for example 0.001 to 1 w / w%, 0.01 to 1 w / w%, 0.05 to 1 w / w%, or 0.1 to 0.5 w / w%, relative to the total weight of the adhesive layer.

[0069] The amount of adhesive (e.g., pressure-sensitive adhesive) in the adhesive layer is not limited to the following, but is typically 70 w / w% or more, preferably 75 w / w% or more, more preferably 75 to 90 w / w%, 75 to 85 w / w%, 80 to 95 w / w%, or 85 to 95 w / w%, 85 to 90 w / w%, for example 80 w / w% or 90 w / w%, relative to the total weight of the adhesive layer.

[0070] In one embodiment, the amount of the non-lamellar liquid crystal forming lipid and the adhesive in the adhesive layer of the patch, for example, the tape, may be, but is not limited to, 1 to 20 w / w% and 75 to 90 w / w%, respectively, based on the total weight of the adhesive layer. In another embodiment, the amount of the non-lamellar liquid crystal forming lipid and the adhesive in the adhesive layer of the patch, for example, the tape, may be, but is not limited to, 1 to 10 w / w% and 85 to 95 w / w%, respectively, based on the total weight of the adhesive layer. In another embodiment, the amount of the non-lamellar liquid crystal forming lipid and the adhesive in the adhesive layer of the patch, for example, the tape, may be, but is not limited to, 10 to 15 w / w% and 75 to 85 w / w%, respectively, based on the total weight of the adhesive layer. In another embodiment, the amounts of non-lamellar liquid crystal forming lipid and adhesive in the adhesive layer of a patch, for example a tape, may be, but are not limited to, 13 to 15 w / w% and 75 to 85 w / w%, respectively, relative to the total weight of the adhesive layer.

[0071] The adhesive layer of the patch, for example, the tape, preferably contains an aqueous medium (e.g., water) together with the non-lamellar liquid crystal forming lipid and the drug. By containing an aqueous medium, the non-lamellar liquid crystal forming lipid forms liquid crystal in the patch, and the drug is encapsulated in the liquid crystal. The aqueous medium is as described above. Alternatively, the patch itself may not contain an aqueous medium, and in that case, when the patch is applied to the surface of a living body, the non-lamellar liquid crystal forming lipid may form liquid crystal on the surface of the living body due to the presence of surrounding water, and the drug may be encapsulated in the liquid crystal. In one embodiment, the aqueous medium may be incorporated into the adhesive layer composition as a liquid containing a drug or other components. The amount of the aqueous medium in the adhesive layer is not limited to the following, but is usually 0.1 w / w% or more, preferably 0.5 w / w% or more, more preferably 1 w / w% or more, for example, 5 w / w% or more, 3 to 30 w / w%, 5 to 10 w / w%, 10 to 30 w / w%, 10 to 25 w / w%, or 13 to 15 w / w% based on the total weight of the adhesive layer. In one embodiment, the weight ratio of the non-lamellar liquid crystal forming lipid to the aqueous medium in the adhesive layer may be preferably 1:5 to 5:1, for example, 1:1 to 10:1, 1:1 to 5:1, 1:1 to 3:1, 1.5:1 to 10:1, 2:1 to 5:1, and in a preferred example, 2:1 to 3:1. The weight ratio of the non-lamellar liquid crystal forming lipid to the adhesive in the adhesive layer may be, for example, 1:2 to 1:20, and is preferably 1:2 to 1:15, 1:3 to 1:10, 1:3 to 1:8, or 1:5 to 1:7.

[0072] The adhesive layer may further contain other components, which are as described above for the topical agent according to the present invention.

[0073] The patch, for example, a tape, may be manufactured using a technique known to those skilled in the art. In one embodiment, a liquid crystal gel is prepared by uniformly mixing a non-lamellar liquid crystal forming lipid, a drug, and an aqueous medium, and optionally other components, and then an adhesive is mixed to prepare an adhesive layer composition, the adhesive layer composition is spread on a liner, the adhesive layer is dried, the adhesive layer is pressed and fixed to a support, and if necessary, cut into a certain size, thereby manufacturing a patch, for example, a tape. Alternatively, a patch, for example, a tape, may be manufactured by preparing a liquid crystal gel as described above, mixing an adhesive to prepare an adhesive layer composition, applying the adhesive layer composition onto a support, drying the adhesive layer, and then laminating a liner to the adhesive layer, and cutting into a certain size as necessary. A patch, for example, a tape, may be manufactured in a size and / or shape suitable for application to an affected area, or may be manufactured in a predetermined size and / or shape, and may be cut into a suitable size and / or shape when used.

[0074] In such patches, for example tapes, drugs are incorporated into liquid crystals formed by non-lamellar liquid crystal forming lipids, and when the patch is applied to a biological surface such as the skin or mucous membrane, it functions to increase the drug permeability through the skin or mucous membrane.

[0075] By applying (applying) the patch, for example the tape, of the present invention to the biological surface, preferably the skin or mucosa, of a subject (e.g., a mammal), the drug contained in the patch can be administered to the subject transdermally or transmucosally. The subject is as described above. The present invention also provides a method for administering or delivering such a drug.

[0076] 5. Aerosols The topical agent according to the present invention may be formulated in the form of an aerosol. The present invention also provides an aerosol containing the topical agent according to the present invention. In the present invention, the aerosol refers to a pharmaceutical preparation in a form in which a drug is ejected by the pressure of a propellant filled together with the drug in the same container. The topical agent according to the present invention formulated in the form of an aerosol comprises a composition containing a non-lamellar liquid crystal forming lipid and a drug, and a propellant. Examples of the propellant include liquefied gas and / or compressed gas. Examples of the liquefied gas include liquefied petroleum gas (LPG) and dimethyl ether (DME). Examples of the compressed gas include carbon dioxide, nitrogen, air, and the like. In the aerosol according to the present invention, the propellant is more preferably liquefied gas, and even more preferably LPG.

[0077] The aerosol agent according to the present invention is preferably a composition containing a non-lamellar liquid crystal forming lipid and a drug, and a propellant filled in a container. The composition containing a non-lamellar liquid crystal forming lipid and a drug is preferably liquid, and is preferably aqueous. In other words, the topical agent according to the present invention formulated in the form of an aerosol agent preferably contains an aqueous medium in addition to the non-lamellar liquid crystal forming lipid, the drug, and the propellant. By containing an aqueous medium, the non-lamellar liquid crystal forming lipid forms a liquid crystal, and the drug is encapsulated in the liquid crystal. Alternatively, the topical agent according to the present invention formulated in the form of an aerosol agent may not contain an aqueous medium, but in that case, the non-lamellar liquid crystal forming lipid in the topical agent forms a non-lamellar liquid crystal with the aqueous medium present at the application site (e.g., moisture in the body or external moisture such as added water), and the drug is encapsulated in the liquid crystal. When the aerosol agent according to the present invention is applied to the surface of a living body, the volatile components in the aerosol agent (solvents such as ethanol, propellants, etc.) evaporate, and non-lamellar liquid crystals can be formed on the surface of the living body. In the aerosol preparation according to the present invention, the composition containing the non-lamellar liquid crystal forming lipid and the drug and the propellant may be mixed in a container or may be separated into a plurality of phases. The non-lamellar liquid crystal forming lipid, the drug, and the aqueous medium are as described above.

[0078] In one embodiment, the amount of non-lamellar liquid crystal forming lipid used in an aerosol preparation is, but not limited to, usually 0.1 w / w% or more, preferably 0.5 w / w% or more, more preferably 1 w / w% or more, for example, 1 to 40 w / w%, 3 w / w% or more, 3 to 40 w / w%, 3 to 20 w / w%, 3 to 15 w / w%, 3 to 10 w / w%, 10 to 30 w / w%, or 5 to 9 w / w%, relative to the total weight of the aerosol raw materials filled in the container, typically the total weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug and the propellant (total weight of the topical preparation; the same applies below).

[0079] In one embodiment, the amount of drug used in the aerosol is, but is not limited to, usually 0.0001 w / w% or more, preferably 0.0005 to 5 w / w%, for example, 0.0005 to 1 w / w%, 0.001 to 5 w / w%, 0.001 to 10 w / w%, 0.001 to 1 w / w%, 0.001 to 0.1 w / w%, 0.001 to 0.05 w / w%, 0.001 to 0.01 w / w%, 0.01 to 0.1 w / w%, 0.1 to 3 w / w%, or 0.1 to 1 w / w%, based on the total weight of the aerosol raw material filled in the container, typically the total weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug, and the propellant.

[0080] In one embodiment, the amount of propellant used in the aerosol is, but is not limited to, usually 40 w / w% or more, preferably 50 w / w% or more, for example, 50 to 90 w / w%, 50 to 85 w / w%, 50 to 80 w / w%, 60 to 85 w / w%, 60 to 80 w / w%, 60 to 70 w / w%, or 65 to 75 w / w%, based on the total weight of the aerosol raw material filled in the container, typically the total weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug, and the propellant.

[0081] In one embodiment, the amount of aqueous medium used in the aerosol formulation is, but not limited to, usually 0.1 w / w% or more, preferably 0.5 w / w% or more, more preferably 1 w / w% or more, for example 3 w / w% or more, 5 w / w% or more, 0.1 to 30 w / w%, 1 to 30 w / w%, 3 to 30 w / w%, 3 to 20 w / w%, 3 to 15 w / w%, 1 to 10 w / w%, 3 to 10 w / w%, 5 to 10 w / w%, or 5 to 9 w / w%, based on the total weight of the aerosol raw material filled in the container, typically the sum of the weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug and the propellant.

[0082] In one embodiment, the weight ratio of the non-lamellar liquid crystal forming lipid to the aqueous medium may be preferably 1:5 to 5:1, 1:2 to 2:1, or 1:1.5 to 1.5:1, for example, 1:1.3 to 1.3:1, or 1:1.1 to 1.1:1, and in one preferred example, is 1: 1. Alternatively, the weight ratio of the non-lamellar liquid crystal forming lipid to the aqueous medium may be 1:1 to 10:1, or 1.5:1 to 5:1, for example, 1:1, 1.5:1, 2:1, 3:1, 4:1, or 5:1.

[0083] The topical preparation according to the present invention formulated in the form of an aerosol may further contain a surfactant, but may not. In one embodiment, the composition comprising the non-lamellar liquid crystal forming lipid and the drug contains a surfactant. The surfactant is as described above. A preferred example of the surfactant used in the aerosol is polyoxyethylene (196) polyoxypropylene (67) glycol (also known as Pluronic). (R) Examples of surfactants include, but are not limited to, F127. In one embodiment, the amount of surfactant used in the aerosol agent is, but is not limited to, usually 0.01 w / w% or more, preferably 0.05 w / w% or more, more preferably 0.1 w / w% or more, for example, 0.05 w / w% to 15 w / w%, 0.1 to 5 w / w%, 0.3 w / w% or more, 0.3 to 10 w / w%, 0.3 to 2 w / w%, 0.3 to 1.5 w / w%, 0.3 to 1 w / w%, or 0.55 to 0.9 w / w%, based on the total weight of the aerosol raw material filled in the container, typically the total weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug and the propellant.

[0084] In one embodiment, the weight ratio of non-lamellar liquid crystal forming lipid to surfactant in the aerosol may be preferably 3:1 to 20:1, or 5:1 to 20:1, for example, 3:1 to 11:1, 7:1 to 17:1, or 8:1 to 15:1, 8:1 to 13:1, 9:1 to 11:1, and in one preferred example, is 10:1.

[0085] The topical preparation according to the present invention formulated in the form of an aerosol preferably further contains ethanol. The composition containing the non-lamellar liquid crystal forming lipid and the drug contains ethanol. By adding ethanol, the skin permeability of the drug is further increased.

[0086] In one embodiment, the amount of ethanol used in the aerosol formulation may be, but is not limited to, 1 w / w% or more, preferably 3 w / w% or more, for example, 5 w / w% or more, 7 w / w% or more, 1 to 30 w / w%, 1 to 20 w / w%, 5 to 60 w / w%, 5 to 30 w / w%, 5 to 25 w / w%, 7 to 20 w / w%, 10 to 20 w / w%, 13 to 50 w / w%, 13 to 20 w / w%, or 15 to 18 w / w%, based on the total weight of the aerosol raw material filled in the container, typically the sum of the weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug and the propellant.

[0087] In one embodiment, the amount of ethanol relative to the total weight (excluding the propellant) of the composition comprising the non-lamellar liquid crystal forming lipid and the drug may be, for example, but is not limited to, 20 to 60 w / w%, or 30 to 50 w / w%.

[0088] In one embodiment, the topical preparation according to the present invention formulated in the form of an aerosol contains a drug, a non-lamellar liquid crystal forming lipid (e.g., an amphiphilic compound represented by general formula (I) or a glycerol fatty acid monoester, preferably mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol or mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol), and a surfactant (preferably Pluronic(R) F127), ethanol, and a propellant (preferably LPG). In this case, the amount of ethanol relative to the total weight of the aerosol raw material filled in the container, typically the total weight of the composition containing non-lamellar liquid crystal forming lipids and a drug, and the propellant, may be as described above, but is preferably 5 to 25 w / w%, 13 to 20 w / w%, or 15 to 18 w / w%. In this case, the amount of the propellant relative to the total weight of the aerosol raw material filled in the container, typically the total weight of the composition containing non-lamellar liquid crystal forming lipids and a drug, and the propellant, is preferably 50 to 85 w / w%, 60 to 80 w / w%, or 60 to 70 w / w%. In one embodiment, the amount of ethanol relative to the total weight of the aerosol raw material filled in the container, typically the total weight of the composition containing non-lamellar liquid crystal forming lipids and a drug, and the propellant, may be 5 to 25 w / w%, and the amount of the propellant may be 60 to 80 w / w%. In one embodiment, the amount of ethanol relative to the total weight of the aerosol raw material filled in the container, typically the sum of the weight of the composition containing the non-lamellar liquid crystal forming lipid and the drug and the propellant, may be 15 to 18 w / w%, and the amount of the propellant may be 60 to 70 w / w%.

[0089] The topical preparation according to the present invention formulated in the form of an aerosol may further contain other components. Such other components are basically contained in a composition containing a non-lamellar liquid crystal forming lipid and a drug. The other components are as described above with respect to the topical preparation according to the present invention.

[0090] The aerosol preparation may be produced using a technique known to those skilled in the art. In one embodiment, the non-lamellar liquid crystal forming lipid, the drug, the aqueous medium, and optionally the surfactant and other components are mixed uniformly, and if necessary, ethanol is added and mixed to prepare a composition, which is then placed in a container as an aerosol concentrate, and then a propellant is filled into the container using a gas filling valve or the like, to produce the aerosol preparation. In another embodiment, the non-lamellar liquid crystal forming lipid, the drug, and optionally the surfactant and other components are mixed uniformly, and if necessary, ethanol is added and mixed to prepare a composition, which is then used as an aerosol concentrate.

[0091] By spraying such an aerosol agent, liquid crystals encapsulating drugs are stably formed and attached to the application site on the biological surface, resulting in excellent drug permeability.

[0092] By applying (spraying) the aerosol according to the present invention to a biological surface, preferably the skin or mucosa, of a subject (e.g., a mammal), the drug contained in the aerosol can be administered to the subject transdermally or transmucosally. The subject is as described above. The present invention also provides a method for administering or delivering such a drug.

[0093] 6. Liquid Crystal Precursor Preparation The topical preparation according to the present invention may be one in which the non-lamellar liquid crystal-forming lipid does not form liquid crystals (non-lamellar liquid crystals). If the topical preparation according to the present invention does not contain an aqueous medium or does not contain an aqueous medium in an amount sufficient for the non-lamellar liquid crystal-forming lipid to form liquid crystals, the non-lamellar liquid crystal-forming lipid will not form liquid crystals in the topical preparation. However, when such a topical preparation is applied to a biological surface in the presence of water, the non-lamellar liquid crystal-forming lipid will form liquid crystals on the biological surface and stably adhere to the biological surface. The present invention also provides a topical preparation according to the present invention in which the non-lamellar liquid crystal-forming lipid does not form liquid crystals, i.e., a liquid crystal precursor preparation.

[0094] The topical preparation according to the present invention, which is a liquid crystal precursor preparation, is preferably a composition that contains a non-lamellar liquid crystal forming lipid and a drug, while not containing an aqueous medium or not containing a sufficient amount of an aqueous medium. The non-lamellar liquid crystal forming lipid, the drug, and the aqueous medium are as described above.

[0095] In one embodiment, the amount of non-lamellar liquid crystal forming lipid used in the liquid crystal precursor formulation is not limited to the following, and may be 10 w / w% or more, preferably 30 w / w% or more or 50 w / w% or more, more preferably 60 w / w% or more, for example 70 w / w% or more, 50 to 99.8 w / w% or more, 60 to 99.8 w / w% or more, 65 to 99.8 w / w%, 65 to 99.5 w / w%, 68 to 99 w / w%, 70 to 99.5 w / w%, or 50 to 90 w / w% or 70 to 90 w / w%, relative to the total weight of the liquid crystal precursor formulation (external preparation).

[0096] In one embodiment, the amount of drug used in the liquid crystal precursor formulation is, but is not limited to, typically 0.0001 w / w% or more, preferably 0.0001 to 10 w / w%, for example, 0.0005 to 5 w / w%, 0.0005 to 1 w / w%, 0.001 to 5 w / w%, 0.001 to 1 w / w%, or 0.001 to 0.1 w / w%, relative to the total weight of the liquid crystal precursor formulation (external preparation).

[0097] The topical preparation according to the present invention, which is a liquid crystal precursor preparation, preferably further comprises a water-soluble polymer. The water-soluble polymer is as described above. Preferred examples of the water-soluble polymer include, but are not limited to, hydroxypropyl cellulose (HPC), such as HPC-SSL, HPC-SL, HPC-L, HPC-M, and HPC-H.

[0098] In one embodiment, the amount of the water-soluble polymer in the liquid crystal precursor formulation is not limited to the following, but may be 0.01 w / w% or more, preferably 0.1 w / w% or more, for example, 0.1 to 10 w / w%, 1 to 5 w / w%, or 0.5 to 2 w / w%, relative to the total weight of the liquid crystal precursor formulation (external preparation).

[0099] The topical preparation according to the present invention, which is a liquid crystal precursor preparation, may or may not further contain ethanol. In one embodiment, the amount of ethanol in the liquid crystal precursor preparation may be, but is not limited to, 1 w / w% or more, preferably 4 w / w% or more, for example, 5 to 40 w / w%, 5 to 30 w / w%, or 10 to 30 w / w%, based on the total weight of the liquid crystal precursor preparation (topical preparation).

[0100] The topical preparation according to the present invention, which is a liquid crystal precursor preparation, may or may not further contain an oil. The oil is as described above. Preferred examples of the oil include, but are not limited to, squalene, squalane, isopropyl myristate, tocopherol, and the like. In one embodiment, the amount of the oil in the liquid crystal precursor preparation may be, but is not limited to, 0.01 w / w% or more, for example, 0.01 to 60 w / w%, 0.1 to 40 w / w%, 1 to 30 w / w%, 1 to 10 w / w%, or 1 to 8 w / w%, based on the total weight of the liquid crystal precursor preparation (topical preparation).

[0101] In one embodiment, the topical preparation of the present invention, which is a liquid crystal precursor preparation, may contain a drug, a non-lamellar liquid crystal forming lipid (e.g., an amphiphilic compound represented by general formula (I) or a glycerin fatty acid monoester, preferably mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol or mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol), a water-soluble polymer (preferably hydroxypropylcellulose), and ethanol.

[0102] The topical preparation according to the present invention, which is a liquid crystal precursor preparation, may further contain other ingredients. The other ingredients are as described above for the topical preparation according to the present invention.

[0103] The liquid crystal precursor formulation may be produced using a technique known to those skilled in the art. The liquid crystal precursor formulation can be produced by thoroughly mixing the above-mentioned raw materials. In one embodiment, the liquid crystal precursor formulation can be produced by thoroughly mixing the non-lamellar liquid crystal forming lipid with a water-soluble polymer, ethanol, and optionally an oil, and adding a drug thereto and thoroughly mixing. Alternatively, the liquid crystal precursor formulation can be produced by thoroughly mixing the drug, ethanol, and optionally a water-soluble polymer and an oil, and adding a non-lamellar liquid crystal forming lipid thereto and mixing. Alternatively, the liquid crystal precursor formulation can be produced by simply thoroughly mixing the non-lamellar liquid crystal forming lipid, the drug, the water-soluble polymer, the oil, and the like.

[0104] When such a liquid crystal precursor formulation is applied to the surface of a living body, in the presence of water it forms liquid crystals encapsulating a drug, which adheres stably to the application site and provides excellent drug permeability.

[0105] The liquid crystal precursor formulation of the present invention can be applied to the skin, but is more preferably applied to the mucosa. By applying the liquid crystal precursor formulation of the present invention to the mucosa, the non-lamellar liquid crystal forming lipids come into contact with water derived from the living body, and the non-lamellar liquid crystal forming lipids self-form liquid crystals (non-lamellar liquid crystals) on the mucosal surface. This allows the liquid crystals encapsulating the drug to stably adhere to the mucosa, resulting in excellent drug mucosal permeability. Therefore, the present invention provides not only a formulation for application to the skin, but also a formulation for application to the mucosa. The topical agent of the present invention, which is a liquid crystal precursor formulation, is particularly preferred for application to the mucosa.

[0106] The liquid crystal precursor formulation according to the present invention can be applied (e.g., sprayed, coated, dropped, etc.) to a biological surface, preferably a mucosa or skin, of a subject (e.g., a mammal), to administer a drug contained in the liquid crystal precursor formulation to the subject transmucosally or transdermally. When the liquid crystal precursor formulation according to the present invention is applied to the skin, it is preferable to apply the formulation to skin that has been previously wetted with an aqueous medium, or to add an aqueous medium to the formulation after application of the formulation to the skin. The subject and the aqueous medium are as described above. The present invention also provides a method for administering or delivering such a drug.

[0107] 7. External preparations containing fine particles The present invention also provides an external preparation comprising microparticles comprising the non-lamellar liquid crystal forming lipid and the drug. The external preparation according to the present invention may be a dispersion such as a dispersion liquid (emulsion). The external preparation according to the present invention may comprise a dispersion of microparticles comprising a non-lamellar liquid crystal forming lipid and a drug, for example, a dispersion liquid (microparticle dispersion liquid). The dispersion, for example, the dispersion liquid, preferably comprises the non-lamellar liquid crystal forming lipid and the drug in a dispersion medium, for example, an aqueous medium such as water, and further comprises a dispersing agent such as a surfactant. The dispersion, for example, the dispersion liquid, may further comprise a solvent such as ethanol and / or an oil component as the case may be.

[0108] In the present invention, the term "microparticles" refers to particles having an average particle size of less than 1 mm. The "microparticles" according to the present invention may be microparticles or nanoparticles. In the present invention, the term "microparticles" refers to particles having an average particle size of 1 μm or more and less than 1 mm. In the present invention, the term "nanoparticles" refers to particles having an average particle size of 1 nm or more and less than 1 μm. In the present invention, the term "dispersion" refers to a dispersion medium containing microparticles in a dispersed state in any dispersion medium. In the present invention, the term "microparticle dispersion" refers to a liquid medium containing microparticles in a dispersed state (e.g., an aqueous medium such as water or a physiologically acceptable aqueous solution such as saline). The terms "microparticle dispersion" and "nanoparticle dispersion" refer to liquid media containing microparticles or nanoparticles in a dispersed state (e.g., an aqueous medium such as water or a physiologically acceptable aqueous solution such as saline). The microparticles according to the present invention, for example, microparticles or nanoparticles, are mainly composed of non-lamellar liquid crystal forming lipids and can contain a drug inside. The microparticles, for example, microparticles or nanoparticles, can be prepared by dispersing a liquid crystal phase. In one embodiment, the microparticles, e.g., microparticles or nanoparticles, can be prepared as a dispersion (emulsion) obtained by dispersing a suspension containing a non-lamellar liquid crystal forming lipid, a drug, an aqueous medium, a dispersing agent such as a surfactant, and optionally other components, by high pressure dispersion, ultrasonic treatment, or the like.

[0109] The non-lamellar liquid crystal forming lipid used in the preparation of microparticles can be any of the non-lamellar liquid crystal forming lipids described above.In one embodiment, the non-lamellar liquid crystal forming lipid can be an amphiphilic compound represented by general formula (I), for example, mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol.In another embodiment, the non-lamellar liquid crystal forming lipid can be a glycerol fatty acid monoester, for example, glyceryl monooleate.

[0110] The microparticles can include any drug as described above, but in one embodiment can be a fat-soluble (lipophilic) drug.

[0111] The "fine particles" according to the present invention may have an average particle diameter of preferably 1 nm or more and less than 1 mm, for example, 1 nm to 500 μm, 10 nm to 500 μm, 50 nm to 500 μm, 10 nm to 1 μm, or 50 nm to 50 μm, although not limited thereto. The nanoparticles according to the present invention may have an average particle diameter of preferably 1 nm to 500 nm, for example, 50 nm to 500 nm, 100 nm to 400 nm, or 100 nm to 300 nm, although not limited thereto.

[0112] The microparticle dispersions described above, e.g., microparticle dispersions such as nanoparticle dispersions, can contain, in addition to the non-lamellar liquid crystal forming lipid and the drug, any other component described above. In one embodiment, the microparticle dispersions, e.g., microparticle dispersions such as nanoparticle dispersions, can contain any of the surfactants described above, e.g., polyoxyethylene(196) polyoxypropylene(67) glycol (also known as Pluronic (R) It is also preferred to include a block copolymer of ethylene oxide and propylene oxide, such as ethylene oxide copolymer F127. In one embodiment, the particulate dispersion, e.g., the nanoparticle dispersion, may include a solvent, such as ethanol. In one embodiment, the particulate dispersion, e.g., the nanoparticle dispersion, may include a surfactant and ethanol. In one embodiment, the particulate dispersion, e.g., the nanoparticle dispersion, may include an oil, as described above, a water-soluble organic compound, as described above, and / or other pharma- ceutical acceptable additives.

[0113] In one embodiment, the weight ratio of non-lamellar liquid crystal forming lipid to dispersing agent in a microparticle dispersion, for example a nanoparticle dispersion, is not limited to the following, but is preferably non-lamellar liquid crystal forming lipid:dispersing agent=1:1 to 100:1, for example 3:1 to 50:1, 3:1 to 10:1, 5:1 to 40:1, 10:1 to 30:1, 10:1 to 25:1, or 15:1 to 25:1.

[0114] In one embodiment, when a solvent such as ethanol is used in a microparticle dispersion, for example a nanoparticle dispersion, the weight ratio of the solvent such as ethanol to the non-lamellar liquid crystal forming lipid is not limited to the following, but may be preferably non-lamellar liquid crystal forming lipid:solvent (e.g., ethanol)=1:10 to 10:1, for example 1:1 to 10:1, 1.5:1 to 5:1, 2:1 to 10:1, or 5:1 to 10:1.

[0115] In one embodiment, the amounts of the non-lamellar liquid crystal forming lipid, the drug, the dispersing agent (such as a surfactant), and the solvent (such as ethanol) may be, for example, 1 to 40 w / w%, 0.001 to 10 w / w%, 0.05 to 15 w / w%, and 1 to 30 w / w%, respectively, based on the total weight of the microparticle dispersion, such as a nanoparticle dispersion. In another embodiment, the amounts of the non-lamellar liquid crystal forming lipid, the drug, the dispersing agent (such as a surfactant), and the solvent (such as ethanol) may be, for example, 10 to 30 w / w%, 0.1 to 3 w / w%, 0.3 to 10 w / w%, and 1 to 20 w / w%, respectively, based on the total weight of the microparticle dispersion, such as a nanoparticle dispersion.

[0116] In one embodiment, the amounts of the non-lamellar liquid crystal forming lipid, the drug, and the dispersing agent (such as a surfactant) may be, for example, 1 to 40 w / w%, 0.001 to 10 w / w%, and 0.05 to 15 w / w%, respectively, based on the total weight of the microparticle dispersion, such as a nanoparticle dispersion. In another embodiment, the amounts of the non-lamellar liquid crystal forming lipid, the drug, and the dispersing agent (such as a surfactant) may be, for example, 10 to 30 w / w%, 0.1 to 3 w / w%, and 0.3 to 10 w / w%, respectively, based on the total weight of the microparticle dispersion, such as a nanoparticle dispersion.

[0117] The topical preparation according to the present invention containing the above-mentioned microparticles or microparticle dispersion may be in any of the above-mentioned dosage forms. In one embodiment, the topical preparation according to the present invention containing the microparticles or microparticle dispersion may be for application to a biological surface of a subject (e.g., a mammal), preferably to the skin or mucosa. In one embodiment, the topical preparation according to the present invention containing the microparticles or microparticle dispersion may be for application to a mucosa (mucosal surface), for example, a nasal preparation. In another embodiment, the topical preparation according to the present invention containing the microparticle dispersion or microparticle dispersion may be a spray such as an aerosol.

[0118] 8. Brain Delivery The topical preparation according to the present invention can be advantageously used for drug delivery into the brain. In one embodiment, the topical preparation according to the present invention for drug delivery into the brain may be a topical preparation containing the above-mentioned microparticles or microparticle dispersion. In one embodiment, the topical preparation according to the present invention for application to mucosa (mucosal surface), for example, the topical preparation according to the present invention which is a nasal preparation, is particularly suitable for drug delivery into the brain. Such topical preparation according to the present invention may be a topical preparation according to the present invention containing the above-mentioned microparticles or microparticle dispersion, or may be a patch, aerosol, or liquid crystal precursor preparation. The topical preparation according to the present invention can efficiently deliver drugs into the brain by transdermal or transmucosal administration, for example, intranasal administration. The topical preparation according to the present invention can greatly improve the efficiency of drug delivery into the brain (particularly including the olfactory bulb, cortex, brainstem, cerebellum, midbrain, and / or hippocampus).

[0119] Therefore, the present invention also provides a method for delivering a drug into the brain, comprising administering the topical agent of the present invention to any of the above subjects, such as humans, transdermally or transmucosally, for example, intranasally.Transdermal administration may be performed using, but is not limited to, a patch, a spray (such as an aerosol), an ointment, or a cream.Transmucosal administration may be performed using a patch, a spray (such as an aerosol), a nasal agent, an oral agent, a suppository, or a vaginal agent.

[0120] The dosage of the external preparation according to the present invention for drug delivery into the brain can be appropriately determined by those skilled in the art depending on the drug. For example, the dosage of the external preparation according to the present invention may be 1 ng to 10 g, for example 10 ng to 100 mg, per 1 kg of subject body weight. For example, the dosage of the external preparation according to the present invention containing a microparticle dispersion may be preferably 1 μL to 500 μL, for example 10 μL to 100 μL, in terms of the amount of the microparticle dispersion, per 1 kg of subject body weight.

[0121] The drug contained in the external preparation for drug delivery into the brain may be any drug that is desired to be delivered into the brain, and may be an organic or inorganic compound, or may be a protein, peptide, amino acid, nucleic acid, etc. The drug may be selected from the above-mentioned drugs. Examples of the drug include, but are not limited to, drugs for treating brain and central nervous system diseases such as Alzheimer's disease, Parkinson's disease, cerebrovascular disorders, and brain tumors, antipsychotic drugs, and anesthetics. EXAMPLES

[0122] The present invention will be described in more detail below with reference to examples, although the technical scope of the present invention is not limited to these examples.

[0123] [Example 1] Synthesis of mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol [ka]

[0124] To a solution of 0.65 g (7.1 mmol) of glycerol and 0.59 g (4.3 mmol) of potassium carbonate in dry N,N-dimethylformamide (3.5 mL), 1.0 g (3.5 mmol) of methyl 5,9,13-trimethyltetradec-4-enoate (methyl tetrahydrofarnesyl acetate) was slowly added dropwise at 80° C. After stirring at 100° C. for 18 hours, 1 M hydrochloric acid was added to the reaction solution, and the solution was extracted with ether. The extract was washed successively with saturated sodium bicarbonate water and saturated saline, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography (ethyl acetate / hexane mixture) to obtain the title compound as a colorless transparent liquid.

[0125] Regarding the obtained compound, 1 The results of H-NMR measurement and viscosity measurement are as follows.

[0126] 1 H-NMR spectrum (300 MHz, CDCl 3 ,TMS)δ:0.80-0.90(m,9H),1.00-1.70(m,15H),1.97(td,J=7.8,17.0Hz,2H),2.13(t,J=6.1Hz,1H,OH), 2.25-2.45(m,4H),2.55(d,J=5.2Hz,1H,OH),3.50-4.00(m,3H),4.10-4.25(m,2H),5.08(t,J=6.7Hz,1H) Viscosity: 0.48Pa·s (shear rate 92 1 / s) The synthesized mono-O-(5,9,13-trimethyltetradec-4-enoyl)glycerol is also referred to as C17MGE, or C17 glycerol ester.

[0127] [Example 2] Synthesis of mono-O-(5,9,13-trimethyltetradecanoyl)glycerol [ka]

[0128] 70g (0.53mol) of 2,2-dimethyl-1,3-dioxolane-4-methanol and 36.7g (266mmol) of potassium carbonate were added to 50.3g (177mmol) of methyl 5,9,13-trimethyltetradecanoate, and the mixture was stirred at 85°C for 3 hours under reduced pressure of 200-250mmHg. During this time, the methanol generated by the reaction was distilled off. The resulting reaction solution was concentrated under reduced pressure (50°C → 210°C, 1.4kPa → 0.38kPa), and then purified by silica gel column chromatography (hexane / ethyl acetate) to obtain 43.0g (63% yield) of methyl 5,9,13-trimethyltetradecanoate (2,2-dimethyl-1,3-dioxolane-4-yl).

[0129] To a solution of 32.7 g (85.0 mmol) of (2,2-dimethyl-1,3-dioxolan-4-yl)methyl 5,9,13-trimethyltetradecanoate in tetrahydrofuran (340 mL), 85 mL of 3 M hydrochloric acid was added at room temperature, and the mixture was stirred at the same temperature for 5 hours. This reaction solution was added to ethyl acetate (300 mL) and saturated sodium bicarbonate water (400 mL) and separated. The organic layer obtained was washed with saturated saline and then dried over magnesium sulfate. The residue obtained by filtration and concentration was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain 28.7 g (yield 98%) of the title compound as a colorless, transparent liquid. Regarding the obtained compound, 1 The results of H-NMR are as follows:

[0130] 1 H-NMR spectrum (270 MHz, CDCl 3 ,TMS)δ:0.7-0.9(m,12H),0.95-1.45(m,16H),1.45-1.75(m,3H),2.34 (t, J=7.4Hz, 2H),3.60(dd,J=5.8,11.5Hz,1H),3.70(dd,J=4.0,11.5Hz,1H),3.94(m,1H),4.15(dd,J=5.9,11.7Hz,1H),4.21(dd,J=4.7,11.7Hz,1H) The synthesized mono-O-(5,9,13-trimethyltetradecanoyl)glycerol is also referred to as saturated C17 glycerol ester.

[0131] [Example 3] Synthesis of mono-O-(5,9,13-trimethyltetradeca-4,8,12-trienoyl)glycerol [ka]

[0132] Under reduced pressure of 200-250 mmHg, 13.9 g (50.0 mmol) of methyl 5,9,13-trimethyltetradeca-4,8,12-trienoate (methyl farnesyl acetate) was gradually added dropwise to a solution of 9.2 g (0.10 mol) of glycerol and 0.28 g (2.0 mmol) of potassium carbonate in dry N,N-dimethylformamide (20 mL) at 85°C, and the mixture was stirred at the same temperature for 3 hours. During this time, methanol generated by the reaction was distilled off. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 150 mL), washed with water, saturated sodium bicarbonate water, and saturated saline (twice), and then dried over magnesium sulfate. The residue obtained by filtration and concentration was purified by silica gel column chromatography (hexane / ethyl acetate = 100:0-0:100) to obtain 8.22 g (49% yield) of the title compound as a colorless, transparent liquid. Regarding the obtained compound, 1 The results of H-NMR measurement and viscosity measurement are as follows.

[0133] 1 H-NMR spectrum (270 MHz, CDCl 3 ,TMS)δ:1.5-1.8(m,12H),1.9-2.1(m,8H),2.1(brs,1H,OH),2.25-2.45(m,4H),2.56(brs,1H,OH),3.59(dd,J=5.6,11.2Hz, 1H),3.68(dd,J=3.6,11.2Hz,1H),3.92(m,1H),4.14(dd,J=6.0,11.6Hz,1H),4.21(dd,J=4.8,11.6Hz,1H),5.02-5.16(m,3H) Viscosity: 0.26 Pa·s (shear rate 92 1 / s) The synthetic mono-O-(5,9,13-trimethyltetradeca-4,8,12-trienoyl)glycerol is also called farnesyl glyceryl acetate.

[0134] [Example 4] Synthesis of mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol [ka]

[0135] Under a reduced pressure of 60-70 mmHg and a nitrogen stream, 28.2 g (80.0 mmol) of methyl 5,9,13,17-tetramethyloctadec-4-enoate was gradually added dropwise to a solution of 23.5 g (255 mmol) of glycerol and 0.55 g (4.0 mmol) of potassium carbonate in dry N,N-dimethylformamide (48 mL) at 80°C, and the mixture was stirred at the same temperature for 3 hours. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 200 mL), washed with water, saturated sodium bicarbonate water, and saturated saline (twice), and then dried over magnesium sulfate. The residue obtained by filtration and concentration was purified by silica gel column chromatography (hexane / ethyl acetate = 100:0-30:70) to obtain 13.3 g (40% yield) of the title compound as a pale yellow transparent liquid. The compound obtained was as follows: 1 The results of H-NMR measurement are as follows.

[0136] 1 H-NMR spectrum (300 MHz, CDCl 3 ,TMS)δ:0.80-0.95(m,12H),1.00-1.70(m,22H),1.85-2.15(m,2H),2.15-2.55(m,4H),3 .53-3.78(m,3H),3.80-4.00(m,1H),4.10-4.25(m,2H),5.08(dd,J=6.9Hz,J=6.9Hz,1H) The synthesized mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol is also referred to as C22MGE, or C22 glycerol ester.

[0137] [Example 5] Synthesis of mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol [ka]

[0138] Under a nitrogen atmosphere, 2.5 g of 5% palladium on carbon was added to a solution of 20.6 g (50.0 mmol) of mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol in ethyl acetate (62 mL). After replacing the nitrogen in the system with hydrogen, the mixture was stirred at room temperature under normal pressure hydrogen atmosphere for 42 hours. After replacing the hydrogen in the system with nitrogen, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (ethyl acetate) to obtain 20.2 g (yield 98%) of the title compound as a colorless transparent liquid. Regarding the obtained compound, 1 The results of H-NMR measurement are as follows.

[0139] 1 H-NMR spectrum (300 MHz, CDCl 3 ,TMS)δ:0.7-0.9(m,15H),0.95-1.75(m,26H),2.13(t,J=6.0Hz,OH),2.34(t,J=7.7Hz,2H),2.56(d,J= 5.1Hz,OH),3.55-3.75(m,2H),3.94(m,1H),4.15(dd,J=6.0,11.7Hz,1H),4.20(dd,J=4.7,11.7Hz,1H) The synthesized mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol is also referred to as saturated C22 glycerol ester.

[0140] [Example 6] Synthesis of mono-O-(5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoyl)glycerol (1) Synthesis of methyl 5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoate (methyl geranylgeranyl acetate) Under a nitrogen atmosphere, a solution of 58.1 g (200 mmol) of 3,7,11,15-tetramethylhexadeca-1,6,10,14-tetraen-3-ol (geranylinalool) and 19 mL (0.15 mol) of trimethyl orthoacetate was added dropwise at 135°C over 8 hours. After stirring for 6 hours at the same temperature, a solution of 5.3 mL (42 mmol) of trimethyl orthoacetate and 0.5 mL (4 mmol) of n-hexanoic acid was added dropwise and stirred for another 2 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (3:1, 300 mL), washed with saturated sodium bicarbonate water (twice) and saturated saline, and then dried over magnesium sulfate. After filtration and concentration, 67.24 g of methyl 5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoate (methyl geranylgeranylacetate) was obtained as a crude liquid product, which was used as it was in the next reaction.

[0141] (2) Synthesis of mono-O-(5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoyl)glycerol [ka]

[0142] Under reduced pressure of 200-250 mmHg, 13.9 g (40.0 mmol) of methyl 5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoate (methyl geranylgeranylacetate) was gradually added dropwise to a solution of 7.4 g (80 mmol) of glycerol and 5.5 g (40 mmol) of potassium carbonate in dry N,N-dimethylformamide (16 mL) at 85°C, and the mixture was stirred at the same temperature for 6 hours. During this time, the methanol generated by the reaction was distilled off. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 200 mL), washed with water, saturated sodium bicarbonate water, and saturated saline (twice), and then dried over magnesium sulfate. The residue obtained by filtration and concentration was purified by silica gel column chromatography (hexane / ethyl acetate = 100:0-0:100) to obtain 5.44 g (yield 33%) of the title compound as a transparent liquid. Regarding the obtained compound, 1 The results of H-NMR measurement and viscosity measurement are as follows.

[0143] 1 H-NMR spectrum (270 MHz, CDCl 3 ,TMS)δ:1.55-1.72(m,15H),1.9-2.2(m,13H),2.27-2.45(m,4H),2.53(brs,1H,OH),3.59(dd,J=5.4,11.4Hz,1H),3 .68(dd,J=3,11.4Hz,1H),3.92(m,1H),4.15(dd,J=6.0,11.6Hz,1H),4.21(dd,J=4.8,11.6Hz,1H),5.05-5.15(m,4H) Viscosity: 0.37 Pa·s (shear rate 92 1 / s) The synthesized mono-O-(5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoyl)glycerol is also called geranylgeranyl glyceryl acetate.

[0144] [Example 7] Preparation of liquid crystal precursor formulation 1. Reagents Rhodamine B (RB) and triamcinolone acetonide (TA) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), and hydroxypropyl cellulose (HPC) was purchased from Nippon Soda Co., Ltd. (Tokyo, Japan). Table 1 shows the structural formula and physicochemical parameters of TA. ClogP, a hydrophobicity index, was measured using Chem Draw Ultra 10.0. (R) (PerkinElmer Informatics, Cambridge, MA, USA).

[0145] [Table 1]

[0146] It is known that the viscosity of HPC increases according to the average number of substituted hydroxyl groups (hydroxypropoxy groups) per cellulose (degree of substitution), with a degree of substitution of 0.2-0.4% being called low and 53.4-77.5% being called high. Five grades of HPC were used: SSL, SL, and L with low degrees of substitution, and M and H with high degrees of substitution. The viscosities of the five types of HPC (2% aqueous solution at 20°C) were 2-2.9 mPa·s for SSL, 3-5.9 mPa·s for SL, 6-10 mPa·s for L, 150-400 mPa·s for M, and 1000-4000 mPa·s for H.

[0147] 2. Preparation of Liquid Crystal Precursor Formulation (1) Formulations containing RB or TA as the drug were prepared as follows. First, HPC was added little by little to a vial containing ethanol. The vial was then placed in a hot bath set at 60°C and stirred overnight to completely dissolve the HPC in the vial, obtaining an ethanol solution containing HPC.

[0148] C17MGE and the HPC-containing ethanol solution obtained above were mixed in a weight ratio of 7:3 and thoroughly stirred for 1 hour. In addition, C17MGE and ethanol were added to a vial, mixed, and thoroughly stirred to prepare a solution without HPC.

[0149] RB was added to the solution prepared as above to a final concentration of 0.001% to prepare RB-containing formulations No. 1 to 6, and TA was added to a final concentration of 0.1% to prepare TA-containing formulations No. 7 to 12. These formulations could also be prepared by adding the drug to the HPC-containing ethanol solution or ethanol in advance and then mixing with C17MGE.

[0150] Furthermore, preparations Nos. 13 to 18 containing C17MGE but not ethanol were prepared by adding a drug and HPC to C17MGE and stirring in the absence of ethanol.

[0151] The thus prepared formulations Nos. 1 to 18 are liquid crystal precursor formulations containing no water.

[0152] Furthermore, as a comparative control, preparations No. 19 to 23 not containing C17MGE were prepared by mixing and stirring water, which is used instead of C17MGE, with ethanol, drug, and HPC. Also, an aqueous solution No. 24 of TA was prepared. The composition ratios (weight ratios) of the prepared preparations Nos. 1 to 24 are shown in Table 2.

[0153] [Table 2] TIFF2025029135000017.tif77170

[0154] 3. Preparation of liquid crystal precursor formulation (2) Formulations Nos. 25 to 36 containing RB or TA were prepared in the same manner as in 2 above, except that glyceryl monooleate (GMO) was used instead of C17MGE in 2 above, in accordance with the composition ratios (weight ratios) in Table 3. In addition, formulations Nos. 37 and 38 containing GMO but not ethanol were prepared by adding the drug and HPC to GMO in the absence of ethanol, followed by heating and stirring.

[0155] [Table 3]

[0156] 4. Preparation of liquid crystal precursor formulation (3) Using oil (squalene) + C17MGE, C22MGE, or oil (IPM or tocopherol) + C22MGE, RB or TA-containing formulations No. 39 to 49 and 79 to 81 were prepared in the same manner as in 2 above, according to the composition ratios (weight ratios) in Table 4.

[0157] [Table 4]

[0158] [Example 8] Preparation characteristic test 1. Spray test Formulations No. 1 to 5, 13, 19 to 23, and 46 prepared in Example 7 were placed in a 5 mL spray vial (No. 2, Maruemu Co., Ltd., Japan) and sprayed once vertically downward from a distance of 3 cm onto a Kimwipe moistened with water. The spray vial used can spray approximately 60 μL with one manual push of purified water. The diameter of the area (spray area) where the formulation was sprayed on the Kimwipe was measured. If the formulation could be sprayed and the diameter of the spray area was 1 cm or more, it was judged to be a mist-like spray, and if it was less than 1 cm, it was judged to be a rod-like spray. If the formulation was not sprayed from the nozzle of the spray vial, it was judged to be unsprayable.

[0159] As a result, when the above-mentioned spray vial was used, all the formulations could be sprayed. Formulations Nos. 1, 19, and 20 were sprayed in a mist form, while Formulations Nos. 2 to 5, 13, 21 to 23, and 46 were sprayed in a rod-like form.

[0160] 2. Liquid crystal structure formation test Using the same 5 mL spray vial as in the spray test, formulations No. 1 to 5, 19, 25, and 26 prepared in Example 2 were sprayed once vertically downward from a distance of 15 cm from the bottom of a glass bottom dish (Matsunami Glass Industry Co., Ltd., Japan) onto 150 μL of purified water dropped into the dish. To determine whether or not liquid crystals were formed, the formulations in contact with the water in the dish were observed in polarizing microscope mode using a digital microscope VHX-5000 (Keyence Corporation, Japan).

[0161] Figure 1 shows the results of observation using a polarizing microscope. Polarized images showing a liquid crystal structure were confirmed for all formulations No. 1 to 5, 25, and 26 containing C17MGE or GMO (Figures 1A to E, G, and H). On the other hand, no polarized images showing a liquid crystal structure were observed for formulation No. 19, which contained HPC but did not contain C17MGE (Figure 1F). It was shown that the formulations containing C17MGE formed a liquid crystal structure in the sprayed water. In addition, the formulations using HPC with a lower degree of substitution had clearer polarized images.

[0162] Furthermore, when the time from spraying to liquid crystal formation (appearance of polarized image) was compared for C17MGE and GMO, under conditions without HPC, it was within 5 seconds for C17MGE formulation No. 1 and within 10 seconds for GMO formulation No. 25, and under conditions with HPC of the same grade SSL, it was within 20 seconds for C17MGE formulation No. 2 and within 90 seconds for GMO formulation No. 26, showing that the liquid crystal formation of the formulations containing C17MGE (Nos. 1 and 2) was significantly faster. Both formulations containing C17MGE and GMO adhered to the mucosal surface quickly after spraying, but the formulation containing C17MGE was more likely to adhere to the mucosal surface more quickly after spraying than the formulation containing GMO, and is thought to be more likely to achieve the release and absorption promotion effect of the active ingredient.

[0163] 3. Small-angle X-ray scattering diffraction When the above liquid crystal structure formation test was performed using formulations No. 1 to 3, 13, 45 to 49 containing C17MGE or C22MGE, and formulations No. 25 to 27 and 37 containing GMO, the formulations sprayed into water became gel-like substances. Small angle X-ray scattering (SAXS) diffraction measurements were performed on these gel-like substances using a small angle X-ray scattering (SAXS) device (Rigaku Corporation, Nano-Viewer) to determine whether they had a non-lamellar liquid crystal structure.

[0164] In the small angle X-ray scattering diffraction of the gel-like substances containing formulations No. 1, 13, 37, and 45 to 47 (Figures 2A and D and Figures 3A to D), at least three scattering peaks were observed, and the peak ratio showed a ratio of 1:√3:2, which is characteristic of reversed hexagonal liquid crystals. Therefore, it was confirmed that these formulations form reversed hexagonal liquid crystals.

[0165] In addition, in the small-angle X-ray scattering diffraction of the gel-like substances containing formulations No. 2, 3, 25 to 27 (Figure 2B, C, and E to G), at least six scattering peaks were observed, and the peak ratios were √2:√3:√4:√8, which are characteristic of cubic liquid crystals belonging to the crystallographic space group Pn3m, and 1:√3:2, which are characteristic of reversed hexagonal liquid crystals. Therefore, it was confirmed that when these formulations were used, cubic liquid crystals belonging to the crystallographic space group Pn3m and reversed hexagonal liquid crystals were mixed to form the mixture.

[0166] In the small-angle X-ray scattering diffraction of the gel-like substances containing formulations No. 48 and 49 (Fig. 3E, F), at least eight scattering peaks were observed, and the peak ratios showed the ratio √3:√8:√11:√12:√16:√19:√24:√27, which is characteristic of reversed cubic liquid crystals belonging to the crystallographic space group Fd3m. Therefore, it was confirmed that these formulations form reversed cubic liquid crystals belonging to the crystallographic space group Fd3m.

[0167] [Example 9] Preparation retention test on mucous membrane WBN / ILA-Ht male hairless rats (body weight 200-250g, 8 weeks old) were intraperitoneally administered a triple anesthesia mixture (medetomidine hydrochloride 0.15mg / kg, midazolam 2mg / kg, butorphanol tartrate 2.5mg / kg) and the back skin was excised. The stratum corneum (horny layer) of the excised back skin (3cm x 3cm) was removed by tape stripping, and the resulting stratum corneum-exfoliated skin (exposed epidermis) was used as a mucosa model.

[0168] Formulations No. 1-6, 13-17, 19-23, 25-28, 30, 39-41, 45, and 46 prepared in Example 7 were sprayed once vertically downward from a distance of 3 cm using a spray vial on a mucosa model fixed on a plate covered with aluminum foil. Immediately after spraying, the area of ​​the formulation attached to the mucosa model (= area immediately after spraying) was measured. 15 seconds after spraying, water at 37°C was continuously flowed on the mucosa model tilted at an angle of 45° from the horizontal at a flow rate of 200 mL / min for 1 hour, and then the area of ​​the formulation attached to the mucosa model (= area after water flow test) was measured. The area immediately after spraying and the area after water flow test were measured using a digital microscope VHX-5000 (Keyence Corporation, Japan) in stereomicroscope mode equipped with a manual XY measurement system VH-M100.

[0169] The reduction rate of the preparation-adhered area after the water flow test was calculated according to the following formula. Reduction rate (%) = (area after water flow test - area immediately after spraying) / area immediately after spraying x 100 When this reduction rate (%) was ≦−50%, it was determined that the preparation sample had detached from the mucosa model.

[0170] The water flow test was performed for each formulation at least three times, and the formulation retention of each formulation was judged by classifying it into four groups according to the following criteria. s: No peeled specimen a: The number of peeled specimens is 40% or less of the total number of specimens. b: The number of peeled specimens is between 40% and 70% of the total number of specimens. c: The number of peeled specimens is 70% or more of the total number of specimens.

[0171] For comparison, the commercially available oral mucoadhesive formulation Aftach was used instead of the formulation prepared in Example 7. (R) (Teijin Pharma Limited, Japan) 1 tablet, or oral mucosal ointment formulation Kenalog (R) (Bristol-Myers Squibb, USA) 60 mg was attached or applied to the mucosa model, and a water flow test was similarly performed. The results are shown in Table 5.

[0172] [Table 5]

[0173] As shown in Table 5, the formulations using C17MGE or C22MGE were well retained even after 1 hour of water flow testing, demonstrating high mucoadhesion. In particular, the formulations containing HPC and C17MGE or C22MGE showed extremely high mucoadhesion (retention), and had higher mucoadhesion than the commercially available formulations. Note that the ethanol solution containing HPC without liquid crystal-forming lipids was completely peeled off in the water flow test.

[0174] [Example 10] Preparation retention test on skin A water flow test was performed in the same manner as in Example 9 using Formulation No. 2 and Formulation No. 46, except that the skin excised from the back of a rat before tape stripping was used as the skin model instead of the mucosal model (stratum corneum-denuded skin).

[0175] As a result, both formulations No. 2 and No. 46 peeled off from the skin when the water flow test was started, but the peeled off gel reattached to the skin and became fixed. It is believed that the reattachment to the skin surface was promoted by the entire formulation coming into contact with water. This result suggests that the formulation of the present invention is prone to peeling off on skin with little moisture (oily surface) until it forms liquid crystals and exhibits bioadhesion, but can exhibit high bioadhesion on skin with sufficient moisture.

[0176] [Example 11] In vitro release test The vertical diffusion cell shown in Figure 4 (cell volume: 6.0 mL, effective diffusion area: 1.77 cm2 6.0 mL of 20% ethanol aqueous solution was added to the receiver cell of a dialysis membrane (cellulose tube 24 / 32, Wako Pure Chemical Industries, Ltd., Japan). (R) (Teijin Pharma Limited, Japan) 200 μL was added, after which the formulation was applied and the in vitro release test of the formulation was started.

[0177] The formulations were formulations Nos. 7 to 12, 18, 24, 31 to 36, 38, 79, 80, and 81 (each 200 mg) prepared in Example 7, each containing 0.2 mg of TA as an active ingredient, and Kenalog. (R) Ointment formulation (200 mg) and Aftach (R) One tablet of the formulation was used. (R) The ointment preparation was applied after wiping off the artificial saliva on the dialysis membrane according to the usage instructions.

[0178] To measure the amount of TA released from the formulation, 500μL of the aqueous solution in the receiver cell was sampled over time and replenished with the same amount of 20% ethanol aqueous solution each time. The inside of the cell was kept at 37℃, and the receiver cell was constantly stirred with a magnetic stirrer. During the test, the vertical diffusion cell was placed in an environment maintained at a high humidity of 90% or more using a humidifier.

[0179] Acetonitrile was added to each sample liquid collected over time at a ratio of 1:1 (v / v), stirred, and centrifuged (21,500×g, 5 min, 4°C) to recover the supernatant. The supernatant was measured by high performance liquid chromatography (HPLC) to determine the TA concentration. The HPLC system and conditions used for this measurement are shown in Tables 6 and 7.

[0180] [Table 6]

[0181] [Table 7]

[0182] Table 8 shows the effective diffusion area of ​​the cell (1.77 cm2) calculated from the measured TA concentration. 2 ) based on the cumulative TA release amount per unit effective release area 4 hours after application of the formulation (μg / cm 2 , the average of three or more tests), and Kenalog (R) The relative release amounts compared to the ointment formulation are shown.

[0183] [Table 8]

[0184] Formulations containing C17MGE, GMO, or C22MGE are commercially available Kenalog (R) The TA release rate was almost the same as that of the ointment formulation. Furthermore, among the formulations containing C17MGE, the formulations containing HPC (No. 8-12, 18) tended to have higher TA release rate than the formulation not containing HPC (No. 7).

[0185] [Example 12] In vitro mucosal permeability test The same vertical diffusion cell as in Example 11 (cell volume: 6.0 mL, effective diffusion area: 1.77 cm 2 6.0 mL of phosphate buffered saline (PBS) at pH 6.75 was added to the receiver cell of the 10-well plate (FIG. 4), and hamster oral mucosa (Syrian type (male, 8 weeks old), Sankyo Labo Service Co., Ltd.) was placed in place of the dialysis membrane used in Example 11. Artificial saliva Salivate was placed on the oral mucosa (donor cell side). (R) (Teijin Pharma Limited, Japan) 200 μL was added, after which the formulation was applied and the measurement was started.

[0186] The formulations were formulations No. 7, 8, 24, 31 to 33, 38, and 80 (200 mg each) prepared in Example 7, each containing 0.2 mg of TA as an active ingredient, and Kenalog. (R) Ointment formulation (200 mg) and Aftach (R) One tablet of the formulation was used. (R) The ointment preparation was applied after wiping off the artificial saliva on the oral mucosa according to the usage instructions.

[0187] To measure the amount of TA permeating through the mucosa from the formulation, 500μL of the aqueous solution in the receiver cell was sampled over time and the same amount of PBS was added each time. The inside of the cell was kept at 37℃, and the receiver cell was constantly stirred with a magnetic stirrer. The vertical diffusion cell was placed in an environment maintained at a high humidity of 90% or more using a humidifier, and the test was performed.

[0188] Acetonitrile was added to each sample liquid collected over time at a ratio of 1:1 (v / v), stirred, and centrifuged (21,500×g, 5 min, 4°C) to recover the supernatant. The supernatant was measured by high performance liquid chromatography (HPLC) to determine the TA concentration. The HPLC system and conditions used for this measurement are shown in Tables 6 and 7.

[0189] Figure 5 shows the mucosal permeation behavior of TA from each evaluation formulation. The vertical axis of the graph is the effective diffusion area of ​​the cell (1.77 cm2) calculated from the measured TA concentration. 2 Cumulative TA mucosal permeation amount per unit effective permeation area (μg / cm 2 ) (average of three or more tests).

[0190] Table 9 shows the cumulative amount of TA permeated per mucosa per effective permeation area per 4 and 8 hours after application of the formulation, and the (R) The relative permeation amount compared with the ointment formulation is shown.

[0191] [Table 9]

[0192] All of the formulations 7, 8, 31-33, 38, and 80 showed high mucosal permeability. In particular, formulations No. 8 and 80, which contain C17MGE or C22MGE and HPC, showed high mucosal permeability compared to the commercially available mucosal formulation (Aftach). (R) , Kenalog (R) ) it showed significantly higher mucosal permeability and significantly increased the mucosal absorption of the active ingredient.

[0193] Furthermore, the relative permeation amount after 4 hours in Preparations Nos. 8 and 80 was greater than the relative permeation amount after 8 hours, demonstrating that the drug could be delivered more effectively in a short period of time.

[0194] These results demonstrate that the formulation containing the liquid crystal-forming lipid of the present invention and a water-soluble polymer such as HPC not only has high mucosal and skin adhesion, but also brings about high drug mucosal permeability and significantly enhances drug mucosal absorption.

[0195] [Example 13] Preparation of spray C17MGE, a liquid crystal forming lipid, and Pluronic F-127 were mixed and mixed for 5 minutes with a vortex mixer. Then, a 790 μg / g aqueous solution of fluorescein sodium (FL-Na) was added, and the mixture was homogenized (8000 rpm, 5 minutes) with a high-speed homogenizer (Polytron PT-3100, KINEMATICA, Switzerland) to prepare a liquid crystal gel (Formulation No. 58). The composition of Formulation No. 58 was liquid crystal forming lipid: Pluronic F-127: FL-Na aqueous solution = 1:0.1:1 (weight ratio). The concentration of FL-Na in Formulation No. 58 was 376 μg / g.

[0196] Ethanol was added to Formulation No. 58 in a ratio of Formulation No. 58:ethanol = 1:0.5 or 1:1 (weight ratio), and the mixture was mixed in a vortex mixer for 5 minutes. The resulting solution was filled into a manual 5 mL spray vial (No. 2, Maruemu Co., Ltd., Japan) to prepare pump sprays No. 50 and No. 51, respectively.

[0197] In addition, formulation No. 50 or formulation No. 51 (solution) was added to an aerosol container (Daizo Co., Ltd., Japan), and a liquefied petroleum gas (LPG) filling valve was attached to fill the container with LPG, a propellant, to prepare aerosol formulations No. 52 and No. 53. The composition of formulation No. 52 was formulation No. 50:LPG = 1.5:4 (weight ratio). The composition of formulation No. 53 was formulation No. 51:LPG = 1:2 (weight ratio).

[0198] In addition, spray preparations Nos. 54 to 57 were prepared with the same compositions and in the same manner as preparations Nos. 50 to 53, except that C22MGE was used instead of C17MGE.

[0199] The FL-Na concentration in formulations No. 50 to 57 is set to be 376 μg / g, the same as formulation No. 58, when all the ethanol and LPG have evaporated after spraying.

[0200] As comparative controls, formulation No. 59 (FL-Na concentration 376 μg / g) was prepared by mixing an FL-Na aqueous solution and ethanol in a weight ratio of 1:1, and formulation No. 60 (FL-Na concentration 376 μg / g) was prepared as a 1 mM FL-Na aqueous solution. The composition ratios (weight ratios) of Preparations Nos. 50 to 60 are shown in Table 10.

[0201] [Table 10]

[0202] [Example 14] Testing the properties of sprays 1. Spray test Aerosol preparations Nos. 52, 53, 56 and 57 prepared in Example 13 were shaken five times immediately before spraying, and then sprayed vertically downward onto a glass surface from a distance of 10 cm.

[0203] For these aerosols, the amount sprayed per second was calculated in 1-second increments over a period of 1 to 5 seconds. The amount sprayed per second (g / sec) was approximately 0.723 g / sec for Formulation No. 52, 0.868 g / sec for Formulation No. 53, 0.726 g / sec for Formulation No. 56, and 0.711 g / sec for Formulation No. 57.

[0204] Formulations No. 52, 53, 56, and 57 were all sprayed as a very fine mist, which caused the ethanol to evaporate instantly, and even when the glass surface was tilted at a 45 degree angle 60 seconds after spraying for 1 second, the sprayed formulations remained on the glass surface without running off.

[0205] On the other hand, when 0.19 g, 0.29 g, 0.19 g, and 0.24 g of pump spray formulations No. 50, 51, 54, and 55, which did not contain LPG, were sprayed, when the glass surface was tilted at 45 degrees after 60 seconds, all four sprayed formulations ran off the glass surface.

[0206] The aerosol preparation of the present invention did not run off from the application site even immediately after spraying, demonstrating that the preparation could be applied extremely effectively.

[0207] 2. Measurement of particle size distribution The spray of the present invention can form nanoparticles immediately after spraying.

[0208] Spray agents No. 50 to 57 were each sprayed into a beaker for 1 second, and the resulting compositions were diluted approximately 1000-fold with distilled water to prepare measurement samples. Using a Zetasizer Nano-ZS (manufactured by Malvern Instruments), the particle size distribution and the zeta potential, which indicates the particle surface charge, were measured by dynamic light scattering.

[0209] The average particle size (nm) (Z-Average), PdI (polydispersity index), and zeta potential (mV) obtained as the average of three measurements are shown in Table 11. All of these showed that the emulsion was in a good condition.

[0210] [Table 11]

[0211] 3. Small-angle X-ray diffraction Aerosol Nos. 52, 53, 56, and 57 were each sprayed into a centrifuge tube for 3 seconds, and then 5 mL of purified water was added to obtain a composition, which was then sealed in a marked tube. Small-angle X-ray scattering diffraction measurements were performed using a small-angle X-ray scattering (SAXS) device (Rigaku Corporation, Nano-Viewer) to determine the non-lamellar liquid crystal structure.

[0212] In the small angle X-ray scattering diffraction of the compositions obtained by spraying formulations No. 52, 56, and 57 (Figures 6A, C, and D), at least three scattering peaks were observed, and the peak ratio showed a ratio of 1:√3:2, which is characteristic of inverted hexagonal liquid crystals. Therefore, it was confirmed that these samples formed inverted hexagonal liquid crystals.

[0213] In addition, in the small angle X-ray scattering diffraction of the composition obtained by spraying formulation No. 53 (Figure 6B), at least eight scattering peaks were observed, and the peak ratios were √2:√3:√4:√8, which are characteristic of cubic liquid crystals belonging to the crystallographic space group Pn3m, and 1:√3:2, which are characteristic of reverse hexagonal liquid crystals. Therefore, it was confirmed that this sample was formed by mixing cubic liquid crystals belonging to the crystallographic space group Pn3m and reverse hexagonal liquid crystals.

[0214] Furthermore, formulation No. 56 was sprayed onto a petri dish for 1 second, and then left for about 3 minutes to obtain a white sample, which was then embedded directly into a pinhole slit and subjected to similar small-angle X-ray scattering diffraction measurements to determine the non-lamellar liquid crystal structure. As a result, at least three scattering peaks were observed, and the peak ratio showed a ratio of 1:√3:2, which is characteristic of inverted hexagonal liquid crystals (Figure 6E). Therefore, it was confirmed that this sample formed an inverted hexagonal liquid crystal.

[0215] These aerosols have been shown to be capable of forming non-lamellar liquid crystalline structures and do so with or without the addition of additional water after nebulization.

[0216] [Example 15] In vitro skin permeability test A triple anesthesia mixture (medetomidine hydrochloride 0.15mg / kg, midazolam 2mg / kg, butorphanol tartrate 2.5mg / kg) was administered intraperitoneally to male WBN / ILA-Ht hairless rats (body weight 200-250g, 8 weeks old). After shaving the abdominal skin, a total of four pieces (2cm x 2cm each) were excised from the left and right sides across the midline, and the subcutaneous fat and blood on the dermis side of each piece were carefully removed with scissors to prepare the rat abdominal skin.

[0217] The same vertical diffusion cell as in Example 11 (cell volume: 6.0 mL, effective diffusion area: 1.77 cm 2 6.0 mL of phosphate buffer (PB) at pH 7.4 was added to the receiver cell of the 10-well plate (FIG. 4), and the above-mentioned rat abdominal skin was placed in place of the dialysis membrane used in Example 11. 1.0 mL of PB was added onto the stratum corneum (donor cell side) and hydrated for 1 hour, after which the formulation was applied and measurement was started.

[0218] As the formulations, formulations Nos. 50 to 53 and 56 to 60 prepared in Example 13 were used. The dosage amounts of the formulations were as follows: Formulation No. 50, 0.218 g (containing 54 μg of FL-Na), No. 51, 0.290 g (containing 54 μg of FL-Na), No. 52, 0.723 g (containing 49 μg of FL-Na) sprayed for 1 second, No. 53, 0.868 g (containing 54 μg of FL-Na) sprayed for 1 second, No. 56, 0.726 g (containing 49 μg of FL-Na) sprayed for 1 second, No. 57, 0.711 g (containing 44 μg of FL-Na) sprayed for 1 second, No. 58, 0.145 g (containing 54 μg of FL-Na), No. 59, 1 mL (containing 376 μg of FL-Na), and No. 60, 1 mL (containing 376 μg of FL-Na).

[0219] To measure the amount of FL-Na permeated through the skin from the formulation, 500μL of the aqueous solution in the receiver cell was sampled over time and the same amount of PB was added each time. The inside of the cell was kept at 32℃, and the receiver cell was constantly stirred with a magnetic stirrer.

[0220] Each sample solution collected over time was centrifuged (21,500 × g, 5 min, 4°C), and the supernatant was measured using a fluorescence spectrophotometer (RF-5300PC: Shimadzu Corporation, Japan) (excitation wavelength: 485 nm, fluorescence wavelength: 535 nm) to determine the FL-Na concentration in the sample solution.

[0221] Figure 7 shows the skin permeation behavior of FL-Na from each formulation. The vertical axis of the graph shows the effective diffusion area of ​​the cell (1.77 cm2) calculated from the measured FL-Na concentration. 2Cumulative FL-Na skin permeation amount per unit effective permeation area (μg / cm 2 ) (average of three or four tests).

[0222] Table 12 shows the cumulative FL-Na skin permeation amount per unit effective permeation area 4 hours and 8 hours after application of the formulation, and the relative permeation amount compared to Formulation No. 60 (control).

[0223] [Table 12]

[0224] Sprays No. 50-53, 56, and 57 all showed significantly higher skin permeability than FL-Na aqueous solution (formulation No. 60), liquid crystal gel (formulation No. 58), and 50% ethanol aqueous solution (formulation No. 59). The cumulative FL-Na skin permeation amount 4 hours after application of pump sprays No. 50 and 51 was 27 times and 184 times higher than that of formulation No. 60, respectively. Furthermore, the cumulative FL-Na skin permeation amount 4 hours after application of aerosols No. 52, 53, 56, and 57 was 104 times, 345 times, 33 times, and 138 times higher than that of formulation No. 60, respectively. The cumulative FL-Na skin permeation amount 8 hours after application also showed a similar trend and was high.

[0225] As shown in Example 14, aerosol preparations Nos. 52, 53, 56, and 57 do not run off from the application site immediately after spraying, and when taken into consideration that the preparations can be applied more effectively than pump spray preparations Nos. 50 and 51 which do not contain LPG, it has been shown that they can be used as even more useful skin permeation enhancing preparations.

[0226] [Example 16] Preparation of tape preparation Liquid crystal forming lipids C17MGE, phytantriol (PHY, Tokyo Chemical Industry Co., Ltd., Japan), or C22MGE and an FL-Na aqueous solution in which FL-Na was dissolved in a phosphate buffer solution (PB) of pH 7.4 were filled in a gas-tight syringe (MS-GAN025, Ito Seisakusho Co., Ltd., Japan) at a weight ratio of 1:1, 2:1, or 3:1, respectively, and mixed uniformly to obtain a liquid crystal gel. The FL-Na concentration of the FL-Na aqueous solution was set so that the FL-Na concentration of the final adhesive layer described below was 10 mM. Since PHY is semi-solid at room temperature, it was used after melting it with a hot stirrer (100 °C, 30 minutes).

[0227] Liquid crystal gel with acrylic adhesive DURO-TAK (R) (387-2516, Henkel, Germany) was added to the adhesive layer at a weight ratio of 80% and mixed using a magnetic stirrer (500 rpm, 5 minutes). This mixture was placed on a silicone-treated polyethylene terephthalate (PET) film liner (film binder). (R) The adhesive layer was placed on the left end (0%) of a No. 75E-0010 BD, Fujimori Kogyo Co., Ltd., Japan, and spread from the left end (0%) to the right end (100%) using a No. 510 Baker-type film applicator (Yasuda Seiki Seisakusho Co., Ltd., Japan) set to a coating thickness of 1 mil (25.4 μm) (Figure 8). The spread adhesive layer was dried for 30 minutes in a room at a temperature of 20 ± 2°C and a humidity of 20 ± 5%, and then further dried for 30 minutes in an incubator at a temperature of 32°C and a humidity of 20 ± 2%. After drying, a 75 μm-thick PET film support (film bina) was applied to the dried adhesive layer using an SN-printing rubber roller No. 3 (Taniguchi Matsuoudo Co., Ltd., Japan). (R) Tape formulations (Formulation Nos. 61 to 69) were prepared by pressing a tape prepared from a 100% glycerin-based sucrose solution (Fujimori Kogyo Co., Ltd., Japan) onto a 100% glycerin-based sucrose solution. Table 13 shows the final composition ratios (weight ratios) in the adhesive layer of Preparations Nos. 61 to 69.

[0228] [Table 13]

[0229] In addition, in accordance with the composition ratio (weight ratio) shown in Table 14, liquid crystal forming lipid (C17MGE) and the above acrylic adhesive (pressure-sensitive adhesive) DURO-TAK in weight ratios of 90%, 70%, and 20% were mixed by the same method as above. (R) and the above acrylic adhesive DURO-TAK (90% by weight) without the incorporation of liquid crystal-forming lipids. (R) A tape formulation No. 73 containing the above was prepared.

[0230] [Table 14]

[0231] [Example 17] Characteristics test of tape preparation 1. Image Analysis In Example 16, after the adhesive layer was dried, tape preparations Nos. 61 to 73 before the PET film backing was pressed against the tape preparation were subjected to normal photography and image analysis using a fluorescent microscope.

[0232] Conventional photographs were obtained by photographing each entire tape preparation from 20 cm above using a digital camera (D5300, Nikon Corporation, Japan).

[0233] Fluorescence microscopy images were obtained by photographing three points from the center of the left edge of the tape preparation, namely, 25% (point 1 in Figure 8), 50% (point 2 in Figure 8), and 75% (point 3 in Figure 8) using a fluorescence microscope (BZ-X700, Keyence Corporation, Japan). The conditions for photographing the fluorescence microscope were: objective lens CFI Plan Apo λ 2x, fluorescence filter GFP (OP-87763 BZ-X filter), excitation wavelength 470 / 40 nm, absorption wavelength 525 / 50 nm, dichroic mirror wavelength 495 nm, and gain +6 dB. The exposure time was 1 / 175 s for preparations containing liquid crystal-forming lipids and 1 / 5 s for preparations not containing lipids.

[0234] DURO-TAK at 80 and 90% by weight (R)The adhesive layers of the tape preparations Nos. 61 to 70, which contained 70% by weight of DURO-TAK, were all uniformly spread. (R) The adhesive layer of tape formulation No. 71, which contained 20% by weight of DURO-TAK, was slightly less uniform than formulations Nos. 61 to 70. (R) and DURO-TAK (90% by weight) without the liquid crystal forming lipid. (R) The adhesive layer of the tape preparation No. 73 containing the above was not uniformly spread in any case.

[0235] From these results, it can be seen that DURO-TAK at a certain weight ratio or higher (R) It was shown that a uniform tape preparation can be prepared by mixing the liquid crystal forming lipid of the present invention with the glycerin (preferably 70% or more, more preferably 80% or more).

[0236] 2. Thickness of adhesive layer The overall thickness of the tape formulation was measured using a hand clipper (Synex Gauge, Techlock Co., Ltd., Japan), and the thickness of the adhesive layer of the tape formulation was calculated by subtracting the thickness of the backing (20 μm) and liner (80 μm) from the measured value.

[0237] The thickness of the adhesive layer at three points from the center of the left end (Figure 8) of Formulations No. 63, 66, and 69, namely 25% (point 1 in Figure 8), 50% (point 2 in Figure 8), and 75% (point in Figure 8), was all 15 ± 5 μm (average value of six measurements). Therefore, it was confirmed that the thickness of the adhesive layer of the above-mentioned tape preparation was uniform.

[0238] 3. Analysis of Phase Images The shape of the tape preparation was observed using a scanning probe microscope (SPM-9700HT, Shimadzu Corporation, Japan) in observation fields of 1 μm x 1 μm and 0.5 μm x 0.5 μm, and then phase observation was performed.

[0239] Shape observation of Formulations No. 63 and 73 revealed smooth surface shapes in both cases. Furthermore, phase observation of these formulations revealed that, as shown in Figure 9, no phase image was observed in Formulation No. 73, whereas a characteristic phase image was observed in Formulation No. 63.

[0240] These results show that the physical properties of the adhesive layer surface differ depending on whether or not liquid crystal-forming lipids are included. (R) It was revealed that the tape formulation containing the above had surface properties characteristic of a structure with a certain degree of regularity.

[0241] [Example 18] In vitro release test The horizontal diffusion cell shown in Figure 10 (cell volume: 3.0 mL, effective diffusion area: 0.95 cm 2 3.0 mL of PB was added to a receiver cell (Kobayashi Glass Co., Ltd.), and the preparation was attached to the donor cell side to start the in vitro release test of the preparation. Preparations Nos. 61 to 69 and 73 were used.

[0242] To measure the amount of FL-Na released from the formulation, 500 μL of the aqueous solution in the receiver cell was sampled over time and the same amount of PB was added each time. The inside of the cell was kept at 32°C and the receiver cell was constantly stirred with a magnetic stirrer.

[0243] Each sample solution collected over time was centrifuged (21,500 × g, 5 min, 4°C), and the supernatant was measured using a fluorescence spectrophotometer (RF-5300PC: Shimadzu Corporation, Japan) (excitation wavelength: 485 nm, fluorescence wavelength: 535 nm) to determine the FL-Na concentration in the sample solution.

[0244] The release behavior of FL-Na from each preparation is shown in Figure 11. The vertical axis of the graph shows the average cumulative FL-Na release rate (%) (average of three or four test values) based on the extraction test, calculated from the measured FL-Na concentration.

[0245] The cumulative FL-Na release rate (%) 1 hour and 4 hours after application of the formulation is shown in Table 15. The cumulative FL-Na release rate (%) was calculated according to the following formula. Cumulative FL-Na release rate (%) = Cumulative FL-Na release amount ÷ FL-Na application amount x 100

[0246] [Table 15]

[0247] Formulations No. 61 to 69 all released FL-Na rapidly over time. In particular, formulations No. 62, 63, 65, and 66 showed a high cumulative release rate 1 hour after application, and reached a release rate of nearly 100% 4 hours after application. In particular, formulations No. 62 and 63 reached a release rate of nearly 100% 1 hour after application, and released FL-Na extremely rapidly.

[0248] On the other hand, from the tape preparation No. 73 that does not contain liquid crystal forming lipid, FL-Na was gradually released, reaching a release rate of about 25% 4 hours after application, after which no further release of FL-Na was observed. This shows that the encapsulated drug cannot be released efficiently unless the liquid crystal forming lipid is added.

[0249] [Example 19] In vitro skin permeability test In accordance with the method described in Example 15, an in vitro skin permeability test was carried out by applying formulations No. 61 to No. 69 to the stratum corneum (donor cell side) of rat abdominal skin set in a vertical diffusion cell (FIG. 4). In addition, a 10 mM FL-Na aqueous solution No. 74, in which FL-Na was dissolved in PB, was used as a control.

[0250] The skin permeation behavior of FL-Na from each formulation is shown in Figure 12. The vertical axis of the graph shows the average cumulative FL-Na skin permeation rate (%) (average of three or four test values) based on the extraction test, calculated from the measured FL-Na concentration.

[0251] The cumulative FL-Na skin permeation rate (%) 1 hour and 4 hours after application of the formulation is shown in Table 16. The cumulative FL-Na release rate (%) was calculated according to the following formula. Cumulative FL-Na release rate (%) = Cumulative FL-Na permeation amount ÷ FL-Na applied amount x 100

[0252] [Table 16]

[0253] The skin permeability of all of the tape preparations No. 61 to 69 was significantly improved compared to the FL-Na aqueous solution No. 74. The skin permeability of the preparations No. 61 to 63 containing C17MGE was very high, even compared to the tape preparations No. 64 to 66 containing PHY. Considering that there was no significant difference in the in vitro release between the tape preparations containing C17MGE or C22MGE and the tape preparations containing PHY in Example 18, the high skin permeability shown by the tape preparations containing C17MGE or C22MGE was a surprising result. Furthermore, the skin permeability of the preparations No. 62 to 63 was improved more than that expected from the content ratio of liquid crystal-forming lipids compared to the preparation No. 61 4 hours after application and thereafter (FIG. 12).

[0254] [Example 20] Preparation of emulsion According to the blending ratio (weight ratio) shown in Table 17, liquid crystal forming lipids C17MGE or glyceryl monooleate (GMO, Rikemal XO-100, NOF Corp.), a drug tranilast (Tokyo Chemical Industry Co., Ltd., Japan), and ethanol (only No. 76) were mixed and dissolved in a water bath at 80°C. (R) F127 (Unilube (R)After adding an aqueous solution of Tranilast (70DP-950B, NOF Corp., or Aldrich P2443) in purified water, the mixture was stirred with a spoon or a vortex mixer to form a suspension. The suspension was then ultrasonicated for 5 minutes at 20% amplitude using an ultrasonic homogenizer (Sonics Vibra-Cell VCX-750, Sonics & Materials, Inc.) to prepare white emulsions No. 75 to 77 containing fine particles. Each of these emulsions was prepared in an amount of 10 g. Although tranilast is known as an antiallergic agent, its therapeutic effect on neurological diseases has also been investigated (US 2011 / 0112187 A1).

[0255] In addition, in an emulsion with 20% lipid, C17MGE was added at 1% of Pluronic (R) The GMOs could be dispersed with F127, but not with the 5% Pluronic. (R) Dispersed in F127 (No. 77). As a control, formulation No. 78 was prepared by adding 0.5% tranilast to saline (Table 17).

[0256] [Table 17]

[0257] [Example 21] Evaluation of emulsion properties The particle size distribution and small angle X-ray scattering diffraction of Emulsions No. 75 to 77 prepared in Example 20, and the viscosity and encapsulation efficiency of tranilast of Emulsions No. 75 and 76 were measured.

[0258] The particle size distribution was measured by dynamic light scattering using a Zetasizer Nano-ZS (Malvern). Measurement samples were prepared by diluting each emulsion 1000 times with distilled water. For each measurement sample, the average particle size (nm) (Z-Average), PdI (polydispersity index), and zeta potential (mV) obtained as the average of three measurements are shown in Table 18.

[0259] All emulsions were stable throughout the experiment with no visible aggregates, as evidenced by the appropriate mean particle size, PdI, and zeta potential.

[0260] For small-angle X-ray scattering diffraction, each emulsion was sealed in a mark tube and measured using a small-angle X-ray scattering (SAXS) device (Rigaku Corporation, Nano-Viewer).

[0261] In the small-angle X-ray scattering diffraction of emulsion No. 75 (Figure 13A), at least three scattering peaks were observed, and the peak ratio was 1:√3:2, which is characteristic of inverted hexagonal liquid crystals. Therefore, this emulsion was shown to be a liquid crystal emulsion (hexasome) in which inverted hexagonal liquid crystal particles were dispersed in the aqueous phase.

[0262] In addition, at least four scattering peaks were observed in the small-angle X-ray scattering diffraction of emulsion No. 76 (Figure 13B), and the peak ratio was √2:√3:√4:√8, which is characteristic of cubic liquid crystals belonging to the crystallographic space group Pn3m. Therefore, this emulsion was shown to be a liquid crystal emulsion (cubosome) in which cubic liquid crystal particles belonging to the crystallographic space group Pn3m were dispersed in an aqueous phase.

[0263] In addition, at least three scattering peaks were observed in the small-angle X-ray scattering diffraction of emulsion No. 77 (Figure 13C), and the peak ratio was √2:√4:√6, which is characteristic of cubic liquid crystals belonging to the crystallographic space group Im3m. Therefore, this emulsion was shown to be a liquid crystal emulsion (cubosome) in which cubic liquid crystal particles belonging to the crystallographic space group Im3m were dispersed in an aqueous phase.

[0264] The viscosity was measured using a viscometer (RE215H; cone rotor 0.8°xR24, Toki Sangyo Co., Ltd.). The viscosity (mPa s) of each emulsion measured at a temperature of 25°C and a rotation speed of 50 rpm is shown in Table 18. All of these viscosities were within the sprayable range.

[0265] To calculate the encapsulation efficiency, each emulsion was centrifuged (21,500×g, 15 min, 4°C), and the resulting supernatant was taken out and diluted 10-fold with acetonitrile, and tranilast was quantified using liquid chromatography-tandem mass spectrometry (LC-MS / MS). The encapsulation efficiency was calculated based on the following formula:

[0266]

number

[0267] In the above formula, %EE, TL total , T.L. free indicate the encapsulation efficiency, the total tranilast concentration in the emulsion, and the tranilast concentration in the supernatant, respectively.

[0268] As shown in Table 18, the encapsulation efficiency of each emulsion was high, indicating a high ability to incorporate tranilast into the liquid crystal structure inside the microparticles.

[0269] [Table 18]

[0270] [Example 22] In vitro release test The vertical diffusion cell shown in Figure 4 (cell volume: 6.0 mL, effective diffusion area: 1.77 cm 2 6.0 mL of phosphate buffered saline (PBS; pH 7.4) was added to the receiver cell of a dialysis device (Kobayashi Glass Co., Ltd.), and a pre-hydrated dialysis membrane (molecular weight cutoff = 12,000-14,000 Da, Sanko Junyaku Co., Ltd., Japan) was set in the receiver cell. The formulation was applied onto the dialysis membrane (donor cell side) to start the in vitro release test of the formulation. 1 mL each of emulsion formulations No. 75 and 76, and comparative formulation No. 78 were used.

[0271] To measure the amount of tranilast released from the formulation, 500 μL of the aqueous solution in the receiver cell was sampled over time and replenished with the same amount of PBS each time. The inside of the cell was kept at 32°C and the inside of the receiver cell was constantly stirred with a magnetic stirrer.

[0272] Acetonitrile was added to each sample solution at a ratio of 1:1 (v / v), stirred, and centrifuged (21,500×g, 5 min, 4°C) to recover the supernatant. 10 μL of the supernatant was injected into the LC / MS / MS system to quantify tranilast.

[0273] The LC / MS / MS system used in this measurement consisted of a system controller (CBM-20A, Shimadzu Corporation), a pump (LC-20AD, Shimadzu Corporation), an autosampler (SIL-20ACHT, Shimadzu Corporation), a column oven (CTO-20A, Shimadzu Corporation), a mass spectrometer (4000QTRAP, AB Sciex Corporation), and analytical software (Analyst® version 1.4.2, Shimadzu Corporation).

[0274] The measurement conditions for LC / MS / MS were as follows: the column (Shodex ODP2HPG-2A 2.0 mm x 10 mm, Showa Denko K.K.) was kept at 40°C. The mobile phase was acetonitrile: 5 mM ammonium acetate aqueous solution containing 0.05% formic acid = 80:20. The flow rate was maintained at 0.2 mL / min. Mass spectrometry quantification was performed in multiple reaction monitoring (MRM) mode, monitoring transition ions from m / z 328.0 to m / z 191.2 with a collision energy of 36 eV.

[0275] Figure 14 shows the release profile of tranilast from each formulation 8 hours after application. The vertical axis of the graph shows the cumulative release amount of tranilast (μmol / cm 2 ) (average of four test values).

[0276] The release behavior of tranilast from each formulation correlated well with the Higuchi equation (Higuchi T., J.Pharm.Sci., 52, 1145-1148(1963)). The release rate of tranilast (μmol / cm ) calculated from FIG. 2 / h 0.5 ) was high at 0.17 for formulation No. 76, and was 0.05 for both No. 75 and No. 78. Furthermore, the cumulative tranilast release rates (%) 8 hours after application of the formulations calculated from Figure 14 were 1.82, 3.67, and 2.35 for No. 75, No. 76, and No. 78, respectively. Formulations No. 75 and No. 76, which have tranilast encapsulated in the liquid crystal structure, showed reliable release properties.

[0277] [Example 23] Pharmacokinetic evaluation by intranasal administration Pharmacokinetic evaluation of formulations No. 75 to 78 by intranasal administration was performed using Sprague-Dawley rats (male, 7 weeks old, body weight 230g ± 10g). First, the rats were given general anesthesia by intraperitoneal administration of a triple-mix anesthesia (medetomidine hydrochloride 0.375mg / kg, butorphanol tartrate 2.5mg / kg, midazolam 2mg / kg). The tip of a micropipette was inserted 0.5cm into the nostril of the rat in a supine position, and 10μL of the formulation was dripped into the nostril for intranasal administration.

[0278] Approximately 200 μL of blood was collected from the jugular vein of each rat at the designated time points (0.17, 0.5, 1, 2, 4, and 8 hours after administration), directly transferred to a heparinized tube, and immediately centrifuged (21,500 × g, 10 min, 4 ° C) to obtain plasma. Each time blood was collected, the rats were injected with the same amount of saline via the tail vein. In some rats, 2, 4, or 8 hours after administration, blood was collected, and then the rats were anesthetized by intraperitoneal administration of a triple-mix anesthesia, and the rats were perfused with cold PBS and the whole brains of the rats were removed. The whole brains were dissected into specific regions (olfactory bulb, cortex, brainstem, cerebellum, midbrain, and hippocampus) on ice. Spinal cords were also collected from the sacrificed rats. The collected brain samples were weighed, then cut into pieces using scissors, 0.5 mL of acetonitrile was added, and the samples were homogenized at 12,000 rpm for 5 min at 4°C using a homogenizer (Polytron PT1200E, KINEMATICA, Switzerland). The brain homogenate was centrifuged (21,500×g, 5 min, 4°C) and the supernatant was collected. The supernatants from the plasma and brain homogenates were kept at -30°C until analysis.

[0279] Acetonitrile was added to 50 μL of the supernatant obtained from the plasma or brain homogenate at a ratio of 1:1 (v / v), and the mixture was stirred and then centrifuged (21,500×g, 5 minutes, 4° C.) to recover the supernatant. In the same manner as in Example 22, 10 μL of the obtained supernatant was injected into the LC / MS / MS system to quantify tranilast.

[0280] The changes in tranilast concentration in plasma and brain over 8 hours after intranasal administration of each formulation are shown in Figures 15 and 16. The vertical axis of the graph shows the average tranilast concentration (ng / mL or ng / g) (average of 3 to 5 test values).

[0281] Based on the results shown in Figures 15 and 16, the pharmacokinetic parameters of tranilast in plasma and brain 8 hours after intranasal administration of each formulation were determined as follows: time to maximum concentration (Tmax), maximum concentration (Cmax), and area under the drug concentration-time curve (AUC 0-8 These values ​​are shown in Table 19.

[0282] [Table 19]

[0283] The Tmax in plasma was 0.17 hours after the first blood draw after intranasal administration for both Emulsions No. 75-77 and Comparative Preparation No. 78, demonstrating rapid systemic absorption. The Tmax in brain was 2 hours after the first blood draw after intranasal administration for Comparative Preparation No. 78, whereas it was 8 hours or more after intranasal administration for all Emulsions No. 75-77.

[0284] In plasma, there was no significant difference in Cmax between emulsions No. 75 and 76 using C17MGE and the control formulation No. 78, but the AUC 0-8 was more than three times higher than that of No. 78. In the brain, the Cmax of No. 75 and No. 76 was approximately eight times higher than that of No. 78, and the AUC 0-8 On the other hand, the Cmax and AUC of emulsion No. 77 using GMO were approximately 15 times higher than those of the control formulation No. 78. 0-8 Although both were lower than those of the control, Cmax and AUC 0-8 Both were about five times higher.

[0285] As described above, in emulsions No. 75 and 76 using C17MGE, high concentrations of tranilast were detected in both the plasma and brain, exceeding the concentration shown in emulsion No. 77 using GMO. It is also noteworthy that the tranilast concentration in the brain was significantly increased in emulsions No. 75 and 76 compared to No. 78. Therefore, it was revealed that emulsions using C17MGE are superior in drug delivery to the brain by intranasal administration.

[0286] The tranilast concentrations in different regions of the brain 2, 4, and 8 hours after intranasal administration of each formulation are shown in Figures 17 and 18. The vertical axis of the graphs shows the average tranilast concentration (ng / g) (average of 3 to 5 test values).

[0287] Emulsions No. 75-77 and control formulation No. 78 showed tranilast uptake in all brain regions starting 2 hours after initial extraction. Among all brain regions, the tranilast concentrations from these formulations in the olfactory bulb and spinal cord were generally higher than those in other brain regions (Figures 17 and 18). Since the olfactory bulb is adjacent to the nasal cavity and the spinal cord is the entrance to the brain for the systemic pathway, these results suggest that tranilast is transported into the brain via both the olfactory pathway and the systemic pathway.

[0288] The tranilast concentrations in all brain regions from emulsions 75 and 76 using C17MGE were generally higher over 8 hours after intranasal administration than emulsion 77 using GMO and comparison formulation 78, and were significantly higher 8 hours after intranasal administration (Figures 17 and 18). This indicates that higher concentrations of tranilast accumulated throughout the brain regions for a longer period of time in emulsions 75 and 76 than in emulsion 77 and comparison formulation 78. [Industrial Applicability]

[0289] According to the present invention, it is possible to provide an external preparation having excellent drug absorbability by the living body. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A liquid composition comprising a non-lamellar liquid crystal-forming lipid, a drug, and an aqueous medium.

2. The liquid composition according to claim 1, further comprising a solvent.

3. The liquid composition according to claim 2, wherein the solvent is ethanol.

4. The liquid composition according to claim 1, comprising a non-lamellar liquid crystal-forming lipid, a drug, and an aqueous medium, which are uniformly mixed.

5. The liquid composition according to any one of claims 1 to 4, wherein the non-lamellar liquid crystal-forming lipid is an amphiphilic compound represented by the following general formula (I) or a salt thereof. 【Chemical 1】 (In the formula, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, m represents 1 or 2, 【Chem.】 represents a single bond or a double bond, and R represents a hydrophilic group having two or more hydroxyl groups)

6. The liquid composition according to claim 5, wherein R in the formula represents a hydrophilic group from which one hydroxyl group has been removed from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol.

7. The liquid composition according to any one of claims 1 to 6, wherein the non-lamellar liquid crystal-forming lipid is mono-O-(5,9,13,17-tetramethyloctadeca-4-enoyl)glycerol or mono-O-(5,9,13-trimethyltetradeca-4-enoyl)glycerol.

8. The liquid composition according to any one of claims 1 to 4, wherein the non-lamellar liquid crystal-forming lipid is phytantriol.

9. The liquid composition according to any one of claims 1 to 8, further comprising a surfactant.

10. The liquid composition according to any one of claims 1 to 9, wherein the drug is a water-soluble drug.

11. An external preparation comprising the liquid composition according to any one of claims 1 to 10.

12. The external preparation according to claim 11, for skin application.

13. The external preparation according to claim 11 or 12, which is a spray.

14. The external preparation according to any one of claims 11 to 13, which is an aerosol further comprising a propellant.

15. The external preparation according to claim 11 or 12, which is a patch.

16. The external preparation according to claim 15, which is a tape preparation comprising the liquid composition and an adhesive layer containing an adhesive.

17. The external preparation according to claim 16, comprising 70 w / w% or more of an adhesive based on the total weight of the adhesive layer.

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