JP2019099478A - Powder particulate formulation having non-lamellar liquid crystal reconstruction capability - Google Patents

Abstract

To provide a powder formulation capable of stably reconstructing a non-lamellar liquid crystal.SOLUTION: A powder particulate formulation having non-lamellar liquid crystal reconstruction capability contains an amphiphilic compound represented by a general formula (I) in the figure. (In the formula, X and Y each represent a hydrogen atom or together represent an oxygen atom; n represents an integer from 0 to 2; m represents 1 or 2; the bond in the figure represent a single bond or double bond; and R represents a hydrophilic group having two or more hydroxy groups.)SELECTED DRAWING: None

Description

  The present invention relates to a powder microparticle preparation having non-lamellar liquid crystal reconstitution ability.

  Lyotropic liquid crystals such as liposomes have been reported to be useful as biomimetic drug delivery system (DDS) carriers in many ways from the time when the DDS concept was proposed. In recent years, non-lamellar liquid crystal (NLLC), which is a type of lyotropic liquid crystal, has advantages such as high drug content, ease of preparation, and high stability in macromolecular drugs as compared to conventional DDS carriers. It has been reported. However, since non-lamellar liquid crystals are non-closed vesicles in which cylindrical pore structures are connected, there is also a problem that the drug for enclosing is released in a relatively short time. In addition, the liquid fine particle preparation containing non-lamellar liquid crystal is liable to cause aggregation and creaming, to be unstable easily, and also has a problem that the enclosed drug tends to leak out during storage. Therefore, development of a more improved preparation using non-lamellar liquid crystals is desired.

  Various liquid crystal-forming lipids are used in various fields such as cosmetics and pharmaceuticals. In recent years, a lipid compound capable of forming a cubic liquid crystal exhibiting high stability even at low temperatures (less than 6 ° C.) has been developed, and the use of that liquid crystal in a sustained release preparation has also been reported (Patent Document 1). However, since these liquid crystal compounds have high viscosity and can not pass through a thin injection needle (for example, 30 gauge), they are difficult to use for injections. Therefore, a lower viscosity lipid compound that stably forms cubic liquid crystal has been developed as a base for injections (Patent Document 2).

  Patent Document 3 reports a dry powder formulation capable of reconstituting cubic liquid crystals. However, the liquid crystal composition reconstituted with this preparation is still desired to be improved in terms of stability and the like.

WO 2006/043705 International Publication No. 2011/078383 U.S. Patent Application Publication No. 2002/016 040

  An object of the present invention is to provide a powder formulation capable of stably reconstituting non-lamellar liquid crystals.

  MEANS TO SOLVE THE PROBLEM As a result of repeating earnestly examining in order to solve the said subject, the present inventors discover that the powder microparticle preparation which can reconfigure | reconstruct the non-lamellar liquid crystal stably in an aqueous medium can be manufactured using predetermined liquid crystal formation lipid. The present invention has been completed.

（式中、Ｘ及びＹはそれぞれ水素原子を表すか又は一緒になって酸素原子を表し、ｎは０〜２の整数を表し、ｍは１又は２を表し、

は一重結合又は二重結合を表し、Ｒは２つ以上の水酸基を有する親水性基を表す）
［２］ 前記式中のＲがグリセロール、エリスリトール、ペンタエリスリトール、ジグリセロール、グリセリン酸、トリグリセロール、キシロース、ソルビトール、アスコルビン酸、グルコース、ガラクトース、マンノース、ジペンタエリスリトール、マルトース、マンニトール、及びキシリトールからなる群から選択されるいずれか１つから１つの水酸基が除かれた親水性基を表す、［１］に記載の粉末微粒子製剤。
［３］ 前記式中のＲがグリセロールから１つの水酸基が除かれた親水性基を表す、［１］又は［２］に記載の粉末微粒子製剤。
［４］ 前記両親媒性化合物が、
モノＯ−（５，９，１３−トリメチルテトラデカ−４−エノイル）グリセロール、
モノＯ−（５，９，１３−トリメチルテトラデカノイル）グリセロール、
モノＯ−（５，９，１３−トリメチルテトラデカ−４，８，１２−トリエノイル）グリセロール、
モノＯ−（５，９，１３，１７−テトラメチルオクタデカ−４−エノイル）グリセロール、
モノＯ−（５，９，１３，１７−テトラメチルオクタデカノイル）グリセロール、及び
モノＯ−（５，９，１３，１７−テトラメチルオクタデカ−４，８，１２，１６−テトラエノイル）グリセロール
からなる群から選択される、［１］〜［３］のいずれかに記載の粉末微粒子製剤。
［５］ 界面活性剤をさらに含む、［１］〜［４］のいずれかに記載の粉末微粒子製剤。
［６］ 界面活性剤が非イオン性界面活性剤である、［５］に記載の粉末微粒子製剤。
［７］ 界面活性剤がポリオキシエチレン（１９６）ポリオキシプロピレン（６７）グリコールである、［５］又は［６］に記載の粉末微粒子製剤。
［８］ 水溶性炭水化物をさらに含む、［１］〜［７］のいずれかに記載の粉末微粒子製剤。
［９］ 水溶性炭水化物がトレハロース又はマンニトールである、［８］に記載の粉末微粒子製剤。
［１０］ 薬理活性物質をさらに含む、［１］〜［９］のいずれかに記載の粉末微粒子製剤。
［１１］ 下記一般式（Ｉ）で表される両親媒性化合物と界面活性剤を含むエマルションを、乾燥及び粉末化することを含む、非ラメラ液晶再構成能を有する粉末微粒子製剤の製造方法。

（式中、Ｘ及びＹはそれぞれ水素原子を表すか又は一緒になって酸素原子を表し、ｎは０〜２の整数を表し、ｍは１又は２を表し、

は一重結合又は二重結合を表し、Ｒは２つ以上の水酸基を有する親水性基を表す）
［１２］ 界面活性剤が非イオン性界面活性剤である、［１１］に記載の方法。
［１３］ 界面活性剤がポリオキシエチレン（１９６）ポリオキシプロピレン（６７）グリコールである、［１１］又は［１２］に記載の方法。
［１４］ 前記エマルションが水溶性炭水化物をさらに含む、［１１］〜［１３］に記載の方法。
［１５］ 水溶性炭水化物がトレハロース又はマンニトールである、［１４］に記載の方法。
［１６］ 前記エマルションが薬理活性物質をさらに含む、［１１］〜［１５］のいずれかに記載の方法。
［１７］ 前記両親媒性化合物と界面活性剤を含む水性媒体を、超音波分散法又は高圧分散法で均質化することにより、前記エマルションを調製することを含む、［１１］〜［１６］のいずれかに記載の方法。
［１８］ 前記エマルションを噴射凍結乾燥法により乾燥及び粉末化する、［１１］〜［１７］のいずれかに記載の方法。 That is, the present invention includes the following.
[1] A powder microparticle preparation having non-lamellar liquid crystal reconstitution ability, containing an amphiphilic compound represented by the following general formula (I).

(Wherein, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, and m represents 1 or 2,

Represents a single bond or a double bond, and R represents a hydrophilic group having two or more hydroxyl groups)
[2] R in the above formula consists of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol The powder microparticle preparation according to [1], which represents a hydrophilic group from which one hydroxyl group has been removed from any one selected from the group.
[3] The powder microparticle preparation according to [1] or [2], wherein R in the above formula represents a hydrophilic group in which one hydroxyl group is removed from glycerol.
[4] The amphiphilic compound is
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 The powder microparticle preparation according to any one of [1] to [3], which is selected from the group consisting of
[5] The powder microparticle preparation according to any one of [1] to [4], which further contains a surfactant.
[6] The powder microparticle preparation according to [5], wherein the surfactant is a nonionic surfactant.
[7] The powder microparticle preparation according to [5] or [6], wherein the surfactant is polyoxyethylene (196) polyoxypropylene (67) glycol.
[8] The powder microparticle preparation according to any one of [1] to [7], further comprising a water-soluble carbohydrate.
[9] The powder microparticle preparation according to [8], wherein the water-soluble carbohydrate is trehalose or mannitol.
[10] The powder microparticle preparation according to any one of [1] to [9], further comprising a pharmacologically active substance.
[11] A method for producing a powder microparticle preparation having non-lamellar liquid crystal reconstitution ability, comprising drying and pulverizing an emulsion containing an amphiphilic compound represented by the following general formula (I) and a surfactant.

(Wherein, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, and m represents 1 or 2,

Represents a single bond or a double bond, and R represents a hydrophilic group having two or more hydroxyl groups)
[12] The method according to [11], wherein the surfactant is a nonionic surfactant.
[13] The method according to [11] or [12], wherein the surfactant is polyoxyethylene (196) polyoxypropylene (67) glycol.
[14] The method according to [11] to [13], wherein the emulsion further comprises a water soluble carbohydrate.
[15] The method according to [14], wherein the water soluble carbohydrate is trehalose or mannitol.
[16] The method according to any one of [11] to [15], wherein the emulsion further comprises a pharmacologically active substance.
[17] The method according to [11] to [16], which comprises preparing the emulsion by homogenizing an aqueous medium containing the amphiphilic compound and a surfactant by an ultrasonic dispersion method or a high pressure dispersion method. The method described in either.
[18] The method according to any one of [11] to [17], wherein the emulsion is dried and powdered by a spray lyophilization method.

  According to the present invention, a powder microparticle preparation capable of stably reconstituting non-lamellar liquid crystal can be produced.

FIG. 1 is a photograph showing an SEM observation image of microparticles in a powder microparticle preparation prepared by the DV method using mannitol as an excipient. White bars in the figure represent 100 μm. FIG. 2 is a photograph showing an SEM observation image of microparticles in a powder microparticle preparation prepared by the DV method using trehalose as an excipient. White bars in the figure represent 100 μm. FIG. 3 is a photograph showing an SEM observation image of microparticles in the powder microparticle preparation prepared by the DS method. The white bars in the figure are 30 μm (FIG. 3A), 100 μm (FIG. 3B), 100 μm (FIG. 3C), 50 μm (FIG. 3D), 50 μm (FIG. 3E), 50 μm (FIG. 3F), 10 μm (FIG. 3G) Represent. FIG. 4 shows the results of small angle X-ray diffraction (SAXS) of the emulsion before pulverization. A: Emulsion No. 2b, B: Emulsion No. 5, C: Emulsion No. 1b. FIG. 5 shows the results of small angle X-ray diffraction (SAXS) of the emulsion before pulverization. A: Emulsion No. 6, B: Emulsion No. 7, C: Emulsion No. 8b. FIG. 6 shows the results of small angle X-ray diffraction (SAXS) of an emulsion reconstituted from a powder microparticle preparation. A: Powdered fine particle preparation No. 15, B: Powdered particulate preparation No. 18, C: powder particulate formulation No. 13. FIG. 7 shows the results of small angle X-ray diffraction (SAXS) of an emulsion reconstituted from a powder microparticle preparation. A: Powdered fine particle preparation No. 19, B: Powdered particulate preparation No. 20, C: powder particulate formulation No. 22. FIG. 8 is a diagram showing the analysis results of the powder microparticle preparation by X-ray diffraction method. FIG. 9 is a diagram showing the analysis results of the powder microparticle preparation by X-ray diffraction method.

  Hereinafter, the present invention will be described in detail.

1. Powdered Particulate Formulation The present invention provides a powdered particulate formulation comprising an amphiphilic compound. The powder microparticle preparation of the present invention has non-lamellar liquid crystal reconstitution ability. In the present invention, “powder microparticle preparation” is a powdery microparticle composition, and can be used for medicines, agricultural chemicals, cosmetics, foods and the like. The powder microparticle preparation may be a dry powder (dry powder). In the present invention, "fine particles" mean particles (nanoparticles) having an average particle size of less than 1 mm.

  In a preferred embodiment, the powder microparticle preparation of the present invention contains an amphiphilic compound and a surfactant. The powder microparticle formulation of the present invention may further comprise an excipient such as a water soluble carbohydrate. The powder microparticle preparation of the present invention may further contain a pharmacologically active substance. The powder microparticle preparation of the present invention may contain other substances such as pharmaceutically acceptable additives (eg, carriers, lubricants, disintegrants, wetting agents, stabilizers, buffers, flavoring agents, preservatives, coloring agents). May be further included.

2. Amphiphilic Compound The present invention relates to a powder microparticle preparation (for example, dry powder preparation), which preferably uses a liquid crystal-forming lipid having an isoprenoid type fatty chain as an amphiphilic compound.
In the present invention, an amphiphilic compound represented by the following general formula (I) can be used as such a liquid crystal-forming lipid.

は一重結合又は二重結合を表す。 In the general formula (I), X and Y each represent a hydrogen atom or together represent an oxygen atom. In 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 is 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.
In the ceremony:

Represents a single bond or a double bond.

  R in the general formula (I) represents a hydrophilic group having two or more hydroxyl groups, and is not limited to, for example, glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose And hydrophilic groups from which one hydroxyl group (OH) has been removed from any one selected from the group consisting of sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol and xylitol . More preferably, R in general formula (I) is a hydrophilic group in which one hydroxyl group (OH) has been removed from glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid or xylose, and glycerol Particularly preferred is a hydrophilic group from which a hydroxyl group (OH) has been removed. The hydrophilic group in which one hydroxyl group (OH) is removed from glyceric acid may be a group in which OH (hydroxy group) contained in the carboxyl group of glyceric acid is removed.

は当該両親媒性化合物が幾何異性体のＥ体（シス体）若しくはＺ体（トランス体）又はそれらの混合物であることを意味する。この表記の意味は後述の一般式（ＩＩ）及び（ＩＩＩ）においても同様である。 In the present invention, the notation in the general formula (I):

Means that the amphiphilic compound is a geometric isomer E (cis) or Z (trans) or a mixture thereof. The meaning of this notation is the same as in the general formulas (II) and (III) described later.

  Examples of the amphiphilic compound represented by the general formula (I) include amphiphilic compounds (polyunsaturated fatty acid esters) represented by the following general formula (II).

In the 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 (preferably, 1 or 2), and m is 1 or 2 Represents
R in the general formula (II) comprises glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, and xylitol It represents a hydrophilic group in which one hydroxyl group (OH) has been removed from any one selected from the group. Preferred examples of R are hydrophilic groups in which one hydroxyl group (OH) has been removed from any one selected from the group consisting of glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid and xylose, more preferably Is a hydrophilic group in which one hydroxyl group (OH) has been removed from glycerol. The hydrophilic group in which one hydroxyl group (OH) is removed from glyceric acid may be a group in which OH (hydroxy group) contained in the carboxyl group of glyceric acid is removed.

  As another example of the amphiphilic compound represented by the general formula (I), an amphiphilic compound represented by the following general formula (III) can be mentioned.

In general 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 is 1 or 2 Represents
R in the general formula (III) represents a hydrophilic group having two or more hydroxyl groups, and is not limited to, for example, glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose And hydrophilic groups from which one hydroxyl group (OH) has been removed from any one selected from the group consisting of sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol and xylitol . Preferred examples of R are hydrophilic groups in which one hydroxyl group (OH) has been removed from any one selected from the group consisting of glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid and xylose, more preferably Is a hydrophilic group in which one hydroxyl group (OH) has been removed from glycerol. The hydrophilic group in which one hydroxyl group (OH) is removed from glyceric acid may be a group in which OH (hydroxy group) contained in the carboxyl group of glyceric acid is removed.

  As still another example of the amphiphilic compound represented by the general formula (I), amphiphilic compounds represented by the following general formula (IV) can be mentioned.

In the general 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 is 1 or 2 Represents
R in the general formula (IV) represents a hydrophilic group having two or more hydroxyl groups, and is not limited to, for example, glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglycerol, xylose And hydrophilic groups in which one hydroxyl group (OH) has been removed from any one selected from the group consisting of sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol and xylitol. Preferred examples of R are hydrophilic groups in which one hydroxyl group (OH) has been removed from any one selected from the group consisting of glycerol, pentaerythritol, erythritol, diglycerol, glyceric acid and xylose, more preferably Is a hydrophilic group in which one hydroxyl group (OH) has been removed from glycerol. The hydrophilic group in which one hydroxyl group (OH) is removed from glyceric acid may be a group in which OH (hydroxy group) contained in the carboxyl group of glyceric acid is removed.

As particularly preferred examples of amphiphilic compounds represented by the general formula (I),
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 Examples include, but are not limited to.

3. Properties of amphiphilic compounds The amphiphilic compounds (lipids) used in the present invention are liquid crystal compounds and can form non-lamellar liquid crystals in an aqueous medium. In the present specification, an aqueous medium containing an amphiphilic compound may be referred to as "amphiphilic compound / water system".

  The non-lamellar liquid crystal formed by the amphiphilic compound used in the present invention is preferably a liquid crystal of type II (water-in-oil) having a hydrophobic group oriented to the outside, and specifically, cubic liquid crystal or More preferably, it is a reverse hexagonal liquid crystal.

  The amphiphilic compounds according to the invention exhibit high stability under a wide range of environmental conditions. For example, the amphiphilic compound according to the present invention is characterized by having an isoprenoid chain as a hydrophobic group, and unlike an amphiphilic compound having a linear fatty chain such as oleic acid as a hydrophobic group, it is resistant to hydrolysis. And the oxidation stability is also relatively high. The amphiphilic compound according to the present invention can also stably form liquid crystal even at low temperatures (6 ° C. or less, preferably 0 ° C. or less) because the temperature range in which the liquid crystal can be obtained is wide and the kraft temperature is low. .

  The analysis of the liquid crystal structure formed by the amphiphilic compound can be performed by a conventional method such as observation with a polarizing microscope or small-angle X-ray scattering (SAXS) measurement.

  For example, in order to confirm the liquid crystal formation, it may be examined to have various liquid crystal structures by X-ray small angle scattering (SAXS) method. First, after an amphiphilic compound / water-based sample of a predetermined concentration is placed in, for example, an X-ray capillary tube made of quartz, the capillary may be sealed with an oxygen burner and subjected to SAXS measurement.

  As a result of the SAXS measurement, the formation of liquid crystal can be confirmed by confirming whether or not the following ratio of scattering peaks (peak interval) specific to each liquid crystal structure is exhibited.

Pn 3m cubic liquid crystal ratio:

Ia3d cubic liquid crystal ratio:

Im3m cubic liquid crystal ratio:

Ratios specific to reverse hexagonal liquid crystals:

  In addition, it is possible to easily determine the space group and the lattice constant by calculating peak values from SAXS data and calculating the ratio of their reciprocals according to a method known to those skilled in the art.

  The amphiphilic compound used in the powder microparticle preparation of the present invention itself exhibits low viscosity. Specifically, the amphiphilic compound used in the powder microparticle preparation of the present invention preferably has a compound itself of 15.0 Pa · s or less, more preferably 11.0 Pa · s or less, as measured at 25 ° C. Preferably, it has a viscosity of 6.0 Pa · s or less. This viscosity can be measured, for example, at a temperature of 25 ° C. using a viscosity / viscoelasticity measuring apparatus (Gemini II, Malvern Co., Ltd.).

  The above amphiphilic compound used for the powder microparticle preparation of the present invention can be synthesized according to the synthetic method described in the description of the examples described later or in WO 2014/178256. Alternatively, the amphiphilic compound represented by the general formula (III) can be synthesized, for example, according to the synthesis method described in International Publication WO 2011/078383. Furthermore, the amphiphilic compound represented by the general formula (IV) can be synthesized, for example, according to the synthesis method described in International Publication WO 2006/043705.

  About the compound synthesize | combined, it is preferable to confirm that the target compound was obtained by usual methods, such as NMR measurement.

4. Surfactant The powder microparticle preparation of the present invention can contain a surfactant in addition to the amphiphilic compound described above. When the powder microparticle preparation of the present invention is produced, for example, by drying and pulverizing an emulsion, it preferably contains the above amphiphilic compound and a surfactant. As an example of the surfactant used in the present invention, a block copolymer of hydrophilic ethylene oxide and hydrophobic propylene oxide (polyoxyethylene polyoxypropylene glycol), polyoxyethylene alkyl ether, polyoxyethylene alkyl ester, polyoxy Nonionic surfactants such as ethylene hydrogenated castor oil may be mentioned. The nonionic surfactant is more preferably one having a molecular weight of 1,000 or more (more preferably 5,000 or more). As a block copolymer of ethylene oxide and propylene oxide, polyoxyethylene (200) polyoxypropylene (70) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol, polyoxyethylene (160) polyoxypropylene ( 30) glycol, polyoxyethylene (120) polyoxypropylene (40) glycol and the like. These block copolymers of ethylene oxide and propylene oxide are commercially available under various names such as Pluronic ^(R) , Poloxamer ^(R) , Unilube ^(R) , Pronone ^(R) and the like. Particularly preferred examples of the surfactant include polyoxyethylene (200) polyoxypropylene (70) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol (alias: Pluronic ^(R) F 127; Unilobe 70 DP-950 B, Poloxamers ^(R) 407) and the like, but not limited thereto. In the present invention, the above amphiphilic compound used in the present invention is not included in the range of the surfactant. The powder microparticle preparation of the present invention may contain one or more of such surfactants.

5. Excipient
The powder microparticle preparation of the present invention may also contain an excipient in addition to the above amphiphilic compound, or the above amphiphilic compound and surfactant. The excipient is preferably pharmaceutically, or a pesticide, cosmetic or food product acceptable. The excipient may, for example, be a water soluble carbohydrate. The water soluble carbohydrate is preferably a sugar alcohol, monosaccharide, disaccharide or polysaccharide. Examples of sugar alcohols include mannitol, sorbitol, maltitol, xylitol, erythritol, palatinose and the like. Examples of monosaccharides include glucose, galactose, mannose, fructose and the like. Examples of disaccharides include trehalose, lactose, sucrose, maltose, cellobiose and the like. Examples of polysaccharides include pectin. In one embodiment, the water soluble carbohydrate is a carbohydrate having 4 or more, preferably 5 or more, more preferably 6 or more, for example 4 to 15, 6 to 10, or 6 to 8 hydroxyl groups, Good. Preferred examples of water soluble carbohydrates are mannitol or trehalose. The powder microparticle preparation of the present invention may contain one or more of such water soluble carbohydrates. Alternatively, the powder microparticle preparation of the present invention may contain other excipients such as starch and lactic acid.

6. Pharmacologically Active Substance The powder microparticle preparation of the present invention may further contain one or more pharmacologically active substances. The pharmacologically active substance may be any drug or active ingredient to be formulated into pharmaceuticals, cosmetics, functional foods and the like. The pharmacologically active substance includes, for example, protein, peptide, amino acid, nucleic acid, anticancer drug, immunosuppressant, analgesic, antiinflammatory drug, antiallergic drug, steroid, antiobesity drug, antidiabetic drug, antibiotic, antifungal agent Agents, antiviral agents, vasodilators, anesthetics, antipsychotics, mucoprotective agents, antihypertensive agents, cardiotonics, antianemic agents, antihyperlipidemic agents, bronchodilators, Alzheimer's disease drugs, chronic obstructive It may be a lung disease (COPD) therapeutic drug, a glaucoma therapeutic drug, a cataract therapeutic drug, an age-related macular degeneration therapeutic drug, a hormone drug, a vaccine, and the like, but is not limited thereto.

  When the powder microparticle preparation is reconstituted into a liquid crystal (emulsion) by incorporating the pharmacologically active substance into the powder microparticle preparation of the present invention, the pharmacologically active substance is incorporated into the liquid crystal, and as a result, the solubility of the pharmacologically active substance Absorbency can be improved. When applied to a target application site such as a diseased site, the pharmacologically active substance is less likely to flow out from the site, resulting in high fixability.

7. Preparation of Powder Microparticle Preparation The powder microparticle preparation of the present invention comprises the amphiphilic compound represented by the above general formula (I), and optionally an excipient such as surfactant, water-soluble carbohydrate and / or pharmacology It can be produced by producing dry fine particles containing other components such as an active substance. In a preferred embodiment, the powder microparticle preparation of the present invention comprises an amphiphilic compound represented by the above general formula (I) and a surfactant, and optionally an excipient such as a water soluble carbohydrate and / or a pharmacologically active substance And the like, can be produced by drying and pulverizing an emulsion containing other components such as The present invention relates to dried microparticles comprising an amphiphilic compound represented by the general formula (I), and optionally, other components such as surfactants, excipients such as water-soluble carbohydrates and / or pharmacologically active substances. The present invention relates to a method for producing a powder microparticle preparation, which comprises producing. In a preferred embodiment, the invention relates to amphiphilic compounds of the general formula (I) and surfactants, and optionally other ingredients such as excipients such as water-soluble carbohydrates and / or pharmacologically active substances. There is also provided a method of producing a powder microparticle preparation having non-lamellar liquid crystal reconstituting ability, comprising drying and pulverizing an emulsion containing the same.

  In the method for producing the powder microparticle preparation of the present invention, typically, the amphiphilic compound represented by the above general formula (I) and a surfactant, and optionally an excipient (for example, water soluble carbohydrate) and An emulsion (emulsion) can be prepared by homogenizing the aqueous medium containing the other ingredients such as and / or the pharmacologically active substance. Homogenization can be carried out using any dispersion method, and mechanical dispersion (mechanical emulsification) using a homogenizer or the like, membrane emulsification, etc. can be used. As a mechanical dispersion method, for example, an ultrasonic dispersion method, a high pressure dispersion method, a stirring dispersion method or the like can be used. In the ultrasonic dispersion method, particles in the liquid can be crushed and emulsified by applying ultrasonic vibration to the liquid using, for example, an ultrasonic homogenizer. In the high pressure dispersion method, particles in the liquid can be crushed and emulsified by, for example, using a high pressure homogenizer to load the liquid with high pressure and disrupting the liquid vigorously. In the stirring and dispersing method, particles in the liquid can be crushed and emulsified by stirring, stirring, or shearing the liquid using, for example, a high-speed rotary homogenizer or a high-speed shear stirrer. In one embodiment of the present invention, the above emulsion is preferably prepared using an ultrasonic dispersion method or a high pressure dispersion method.

  In the method for producing a powder microparticle preparation according to the present invention, drying and pulverization of the emulsion may be performed simultaneously, may be dried after pulverization, or may be powdered after drying. The powder microparticle preparation of the present invention can be produced, for example, using any method that can be used to dry and pulverize a liquid composition, such as a lyophilization method, a spray drying method, a spray lyophilization method, and a hot air drying method. In a particularly preferred embodiment, the powder microparticle preparation of the present invention can dry and pulverize the above-mentioned emulsion by spray freeze-drying method. "Spray freeze-drying method" refers to a method of producing freeze-dried microparticles by freeze-drying microdroplets generated by spraying a liquid. In the present invention, “spray” refers to any technique for finely atomizing and spraying a liquid (for example, in the form of a mist), and is not limited by the type of equipment used therein. Spraying in the spray lyophilization method and the like can be performed using any spray means, for example, a spray vial or a single fluid nozzle that sprays (sprays) the liquid alone using a pump or the like, or a gas (compressed air, not Although it can carry out using a two fluid nozzle etc. which mix active gas, steam, etc. with a liquid, and spray it (spray), it is not limited to these methods. In the spray freeze-drying method, for example, the powder microparticle composition is produced by spraying the above-mentioned emulsion against liquid nitrogen and drying the obtained suspension with a freeze-dryer such as a vacuum freeze-dryer. Can. The obtained powder microparticle composition can be used as the powder microparticle preparation of the present invention or as an active ingredient of the powder microparticle preparation. By using the spray freeze-drying method, it is possible to produce a powder microparticle preparation at a low temperature, and the particle size of the microparticles contained in the powder microparticle preparation can be made smaller, and a high recovery rate (typically Almost 100%) can be achieved. In one embodiment, the emulsion can be dried and powdered by spray lyophilization using a two-fluid nozzle.

  In one embodiment of the present invention, the emulsion obtained by the ultrasonic dispersion method may be spray freeze dried using a spray vial or a single fluid nozzle. In another embodiment of the present invention, the emulsion obtained by the high pressure dispersion method may be spray freeze dried using a two fluid nozzle. However, the production of the powder microparticle preparation of the present invention is not limited to these methods, and various dispersion methods can be used in combination with drying and powdering techniques.

  The aqueous medium used in the present invention is not particularly limited, and water such as sterile water, purified water, distilled water, ion exchanged water, ultrapure water, etc .; physiological saline, sodium chloride aqueous solution, calcium chloride aqueous solution, An aqueous electrolyte solution such as an aqueous solution of magnesium chloride, an aqueous solution of sodium sulfate, an aqueous solution of potassium sulfate, an aqueous solution of sodium carbonate, an aqueous solution of sodium acetate, etc .; The aqueous medium used in the present invention may be added with a pharmaceutically acceptable water-soluble organic compound such as ethanol, glycerin, ethylene glycol, butylene glycol and the like. The aqueous medium used to prepare the emulsion of the present invention is preferably physiologically acceptable water or an aqueous solution.

  The amphiphilic compound represented by the general formula (I) can be obtained by mixing the mixture solution used in the above emulsion or the preparation of the above emulsion (i.e., the amphiphilic compound represented by the general formula (I) and surfactant, and 1 to 30%, preferably 1 to 20%, more preferably 5 to 15%, still more preferably 7 to 13% in an aqueous medium containing an excipient and / or other components such as pharmacologically active substances). It is preferred to use in amounts.

  The surfactant is a mixture liquid used in the above emulsion or in the preparation of the above emulsion (i.e. the amphiphilic compound represented by the general formula (I) and the surfactant, and optionally the excipient and / or the pharmacologically active substance And the like, preferably in an amount of 0.1 to 10%, preferably 0.1 to 5%, more preferably 0.5 to 3%, in an aqueous medium containing other components such as

  An excipient, for example, a water-soluble carbohydrate, is used in the above emulsion, or a mixture used for preparing the above emulsion (ie, amphiphilic compound represented by the general formula (I) and surfactant, and optionally an excipient and And / or 1 to 30%, preferably 1 to 20%, more preferably 5 to 15%, still more preferably 7 to 13% in an aqueous medium containing other components such as pharmacologically active substances Value) is preferred.

Also, the above-mentioned emulsion, or a mixed solution used for preparing the above-mentioned emulsion (i.e. amphiphilic compound and surfactant represented by the general formula (I), and optionally other components such as excipient and / or pharmacologically active substance etc. The aqueous medium) preferably contains, for example, 50 to 99%, preferably 60 to 90%, more preferably 70 to 80% of water.
In the present specification,% means weight / weight% unless otherwise stated.

  Specifically, the powder microparticle preparation of the present invention can be produced, for example, as follows. First, the amphiphilic compound represented by the above general formula (I) can be added to the aqueous medium and mixed. The surfactant may be dissolved in the aqueous medium prior to the addition of the amphiphilic compound, or may be added simultaneously with or after the amphiphilic compound. The excipient may be dissolved in the aqueous medium prior to the addition of the amphiphilic compound, or may be added simultaneously with or after the amphiphilic compound. Optionally, other components such as pharmacologically active substances may be further mixed into the obtained mixed solution. Alternatively, the pharmacologically active substance may be dissolved in the aqueous medium prior to the addition of the amphiphilic compound, or may be added to the aqueous medium simultaneously with or after the amphiphilic compound.

  An emulsion can be prepared by homogenizing (dispersing and emulsifying) the mixture thus obtained using a dispersing means such as a high-pressure homogenizer or an ultrasonic homogenizer. This emulsion is a non-lamellar liquid crystal composition containing a non-lamellar liquid crystal structure composed of an amphiphilic compound represented by the general formula (I). Subsequently, dried and powdered fine particles can be suitably produced by, for example, spray freeze-drying the prepared emulsion (non-lamellar liquid crystal composition). Spray lyophilization of the emulsion is carried out, for example, by injecting the emulsion into liquid nitrogen using a commercially available injection device such as a spray vial or a one-fluid nozzle or a two-fluid nozzle, and thereafter evaporating and drying the liquid nitrogen. , Can be done efficiently.

  The size and surface condition of the particles contained in the powder microparticle preparation of the present invention can be controlled by the operation of powdering. For example, when the powder microparticle preparation of the present invention is produced by a spray freeze-drying method using a two-fluid nozzle, the particle diameter of the particles contained in the preparation can be smaller and the particles can be more porous. In addition, by the spray freeze-drying method using a two-fluid nozzle, the particle surface roughness in the powder microparticle preparation of the present invention can be increased, the interparticle adhesion can be reduced, and the flowability of particles can be improved.

  However, the method for producing the powder microparticle preparation of the present invention is not limited to the method based on drying and pulverizing the emulsion. In another embodiment, the powder microparticle preparation of the present invention comprises, for example, adhesion, introduction or encapsulation of an amphiphilic compound represented by the general formula (I) to powder microparticles prepared using an excipient, and then dried It may be manufactured by

  The powder microparticle preparation of the present invention comprises the amphiphilic compound represented by the above general formula (I), and optionally, other components such as surfactants, excipients such as water-soluble carbohydrates and / or pharmacologically active substances Other than pharmaceutically acceptable additives (eg, carriers, lubricants, disintegrants, wetting agents, stabilizers, buffers, flavoring agents, preservatives, coloring agents), etc. The substance may be further added to produce.

  The above-mentioned method for producing the powder microparticle preparation is characterized in that glyceryl monooleate (GMO) or phytantriol (alias: 3, 7, 11, 15) is used instead of the amphiphilic compound represented by the above general formula (I). It may be applied to other liquid crystal forming lipids such as tetramethylhexadecane-1,2,3-triol). Also when other liquid crystal-forming lipids are used, a powder microparticle preparation having non-lamellar liquid crystal reconstituting ability can be produced.

  The powder microparticle preparation of the present invention thus obtained has the ability to reconstitute non-lamellar liquid crystals. In the present invention, "non-lamellar liquid crystal reconfigurability" refers to the ability to form non-lamellar liquid crystals when the powder microparticle preparation of the present invention is dispersed in an aqueous medium. In order to disperse the powder microparticle preparation of the present invention in an aqueous medium, the powder microparticle preparation of the present invention may be added to an aqueous medium. By contacting the powder microparticle preparation of the present invention with an aqueous medium, non-lamellar liquid crystals are spontaneously formed. In order to obtain a more uniform composition, the dispersion may be promoted by stirring or the like for the aqueous medium to which the powder microparticle preparation of the present invention is added to prepare an emulsion. For the purpose of determination of non-lamellar liquid crystal reconstitution ability, the powder microparticle preparation (for example, 50 mg) of the present invention may be mixed with water (for example, 0.2 mL) and stirred with a vortex mixer. For example, after the powder microparticle preparation of the present invention is dispersed in water, the non-lamellar liquid crystal is formed by the liquid crystal structure analysis method as described above, whether the powder microparticle preparation of the present invention can reconstitute the non-lamellar liquid crystal. You can check by checking if it is. In one embodiment, the powder microparticle preparation of the present invention is preferably capable of reconstituting, in an aqueous medium, the same type of non-lamellar liquid crystal as the pre-powdered emulsion. More preferably, the powder microparticle preparation of the present invention can form a reverse hexagonal liquid crystal in an aqueous medium. In a preferred embodiment, an emulsion (liquid crystal emulsion) reconstituted from the powder microparticle preparation of the present invention can form a liquid crystal in which the size of the unit cell does not change significantly as compared to the liquid crystal in the pre-powdering emulsion. That indicates that the powder microparticle preparation of the present invention has the ability to stably maintain the liquid crystal structure of the emulsion before pulverization.

  Emulsions reconstituted from the powder microparticle formulations of the invention usually comprise particles having an average particle size of less than 10 μm, typically from 1 nm to 800 nm. The emulsion preferably has an average particle size of 50 nm to 800 nm, more preferably 50 nm to 600 nm, even more preferably 50 nm to 400 nm, for example 100 nm to 300 nm, or 50 nm to 250 nm. The average particle diameter here is Z average (Z-Average).

  Emulsions reconstituted from the powder microparticle formulations of the invention preferably exhibit a low polydispersity index (PDI). PDI is not limited to the following, but is typically 0.01 or more, preferably 0.5 or less, more preferably 0.4 or less, still more preferably 0.3 or less, for example 0.2 It may be the following.

  The zeta potential is used as an indicator of the dispersion stability of the emulsion. Generally, the larger the absolute value of the zeta potential, the higher the dispersion stability of the particles in the aqueous medium. Since the powder microparticle preparation of the present invention is mainly intended to be administered without being dissolved in an aqueous medium, or to be dissolved in an aqueous medium immediately before administration, it is necessary to have an extremely large zeta potential There is no. However, the powder microparticle preparation of the present invention exhibits a zeta potential of preferably -10 mV or less (e.g. -50mV to -10mV) after reconstitution in an aqueous medium, which means that the preparation was in contact with the aqueous medium In this case, it means that it is possible to produce a liquid crystal composition in a sufficiently redispersed state.

  A liquid crystal emulsion in which fine particles of non-lamellar liquid crystal containing a pharmacologically active substance are dispersed is prepared by dispersing the powder microparticle preparation of the present invention containing the pharmacologically active substance in an aqueous medium to reconstitute non-lamellar liquid crystal. it can. Since such non-lamellar liquid crystals release the pharmacologically active substance slowly, the reconstituted emulsion (non-lamellar liquid crystal composition) can be advantageously used as a sustained release preparation. The emulsion (non-lamellar liquid crystal composition) reconstituted from the powder microparticle preparation of the present invention has a skin / mucosal permeability promoting effect on the pharmacologically active substance to be contained.

  The powder microparticle preparation of the present invention can be used as one component of medicines, agricultural chemicals, cosmetics, foods and the like. Therefore, medicines, agricultural chemicals, cosmetics, foods and the like are also provided, which contain the powder microparticle preparation of the present invention.

  The powder microparticle preparation of the present invention can be directly administered to the application site (for example, affected area) of the living body in the dry state. In this case, the powder microparticle preparation is dispersed in water (such as body fluid) present at the application site or water (such as sprayed water) applied to the application site, whereby the non-lamellar liquid crystal composition is reconstituted at the application site. Be done. Alternatively, the powder microparticle preparation of the present invention may be redispersed in an aqueous medium just before administration, and then administered to the application site (for example, the affected area) of a living body. However, the administration method of the powder microparticle preparation of the present invention is not particularly limited as long as the non-lamellar liquid crystal composition is reconstituted at the time of administration, after administration or immediately before administration.

  The powder microparticle preparation of the present invention can be used to manufacture a preparation of any dosage form intended to reconstitute a non-lamellar liquid crystal composition at the time of administration, after administration or immediately before administration. The powder microparticle preparation of the present invention is, for example, granules, sprays (sprays), inhalants, capsules, tablets (solids), semisolids, ointments, solutions, patches, injections, implants, eye drops It can be formulated into a dosage form such as an agent, suppository, vaginal suppository, etc., but not limited thereto. The powder microparticle formulation of the present invention formulated into any dosage form can also be administered according to the above-described administration method. In one embodiment, certain dosage forms (eg, enteric capsules and patches) can also be used to deliver the powder microparticle formulations of the present invention to the intended site of application.

  Examples of the application site of the powder microparticle preparation of the present invention include, in addition to skin, mucous membranes such as oral cavity, lung, eye, abdominal cavity, intestinal tract, etc., intradermal / subcutaneous, muscle, vein (blood vessel), etc. .

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

5,9,13-trimethyltetradeca-4 at 80 ° C. in 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 -enoate (methyl tetrahydrofarnesyl acetate) was slowly added dropwise. After stirring at 100 ° C. for 18 hours, to the reaction mixture was added 1 M hydrochloric acid, and the mixture was extracted with ether. The extract was washed successively with saturated aqueous sodium bicarbonate solution and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The obtained residue was purified by silica gel column chromatography (ethyl acetate / hexane mixed solution) to give the title compound as a colorless transparent liquid.
The results of ¹ H-NMR measurement and viscosity measurement of the obtained compound are as follows.

¹ H-NMR spectrum (300 MHz, CDCl ₃ , TMS) δ: 0.80 to 0.90 (m, 9 H), 1.00 to 1.70 (m, 15 H), 1.97 (td, J = 7) .8, 17.0 Hz, 2 H), 2.13 (t, J = 6.1 Hz, 1 H, OH), 2.25-2.45 (m, 4 H), 2.55 (d, J = 5. 2 Hz, 1 H, OH), 3.50-4.00 (m, 3 H), 4. 10-4. 25 (m, 2 H), 5.08 (t, J = 6.7 Hz, 1 H) Viscosity: 0 .48 Pa · s (shear rate 92 1 / s)
The synthesized mono O- (5,9,13-trimethyltetradeca-4-enoyl) glycerol is also referred to as C17 glycerol ester.

Example 2 Synthesis of mono O- (5,9,13-trimethyltetradecanoyl) glycerol

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

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

¹ H-NMR spectrum (270 MHz, CDCl ₃ , TMS) δ: 0.7-0.9 (m, 12 H), 0.95 to 1.45 (m, 16 H), 1.45 to 1.75 (m) , 3H), 2.34 (t, J = 7.4 Hz, 2 H), 3.60 (dd, J = 5.8, 11.5 Hz, 1 H), 3.70 (dd, J = 4.0, 11.5 Hz, 1 H), 3.94 (m, 1 H), 4. 15 (dd, J = 5.9, 11.7 Hz, 1 H), 4.21 (dd, J = 4.7, 11.7 Hz) , 1H)
The synthesized mono O- (5,9,13-trimethyltetradecanoyl) glycerol is also referred to as saturated C17 glycerin ester.

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

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

¹ H-NMR spectrum (270 MHz, CDCl ₃ , TMS) δ: 1.5-1.8 (m, 12 H), 1.9-2.1 (m, 8 H), 2.1 (brs, 1 H, OH) ), 2.25 to 2.45 (m, 4 H), 2.56 (brs, 1 H, OH), 3.59 (dd, J = 5.6, 11.2 Hz, 1 H), 3.68 (dd , J = 3.6, 11.2 Hz, 1 H), 3.92 (m, 1 H), 4. 14 (dd, J = 6.0, 11.6 Hz, 1 H), 4.21 (dd, J = 4.8, 11.6 Hz, 1 H), 5.02-5.16 (m, 3 H)
Viscosity: 0.26 Pa · s (shear rate 92 1 / s)
The synthesized mono O- (5,9,13-trimethyltetradeca-4,8,12-trienoyl) glycerol is also referred to as farnesyl glyceryl acetate.

Example 4 Synthesis of Mono O- (5,9,13,17-Tetramethyloctadec-4-enoyl) glycerol

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) under reduced pressure of 60 to 70 mmHg and nitrogen flow at 5, 9, 13 at 80 ° C. Then, 28.2 g (80.0 mmol) of methyl 17-tetramethyloctadec-4-enoate were gradually added dropwise and 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 aqueous sodium bicarbonate solution and saturated brine (twice), and then dried over magnesium sulfate. The residue obtained by filtration and concentration is purified by silica gel column chromatography (hexane / ethyl acetate = 100: 0 to 30:70) to give 13.3 g (yield 40%) of the title compound slightly yellow Obtained as a clear liquid. The results of ¹ H-NMR measurement of the obtained compound are as follows.

¹ H-NMR spectrum (300 MHz, CDCl ₃ , TMS) δ: 0.80 to 0.95 (m, 12 H), 1.00 to 1.70 (m, 22 H), 1.85 to 2.15 (m) , 2H), 2.15 to 2.55 (m, 4H), 3.53 to 3.78 (m, 3H), 3.80 to 4.00 (m, 1 H), 4.10 to 4.25. (M, 2H), 5.08 (dd, J = 6.9 Hz, J = 6.9 Hz, 1 H)
The synthesized mono O- (5,9,13,17-tetramethyloctadeca-4-enoyl) glycerol is also referred to as C22 glycerin ester.

Example 5 Synthesis of Mono-O- (5,9,13,17-Tetramethyloctadecanoyl) glycerol

Under nitrogen atmosphere, 2.5 g of 5% palladium carbon in 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) Added. After replacing the nitrogen in the system with hydrogen, the system was stirred at room temperature for 42 hours under a hydrogen atmosphere. After replacing hydrogen in the system with nitrogen, 5% palladium carbon was filtered off. The filtrate was purified by silica gel column chromatography (ethyl acetate) to give 20.2 g (yield 98%) of the title compound as a colorless, transparent liquid. The results of ¹ H-NMR measurement of the obtained compound are as follows.

¹ H-NMR spectrum (300 MHz, CDCl ₃ , TMS) δ: 0.7-0.9 (m, 15 H), 0.95-1.75 (m, 26 H), 2.13 (t, J = 6) .0 Hz, OH), 2.34 (t, J = 7.7 Hz, 2 H), 2.56 (d, J = 5.1 Hz, OH), 3.55-3.75 (m, 2 H), 3 .94 (m, 1 H), 4. 15 (dd, J = 6.0, 11.7 Hz, 1 H), 4. 20 (dd, J = 4.7, 11.7 Hz, 1 H)
The synthesized mono O- (5,9,13,17-tetramethyloctadecanoyl) glycerol is also referred to as saturated C22 glycerin ester.

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) 3,7,11,15-tetramethylhexadeca under nitrogen atmosphere In a solution of 58.1 g (200 mmol) of 1,6,10,14-tetraen-3-ol (geranyl linalool), 19 mL (0.15 mol) of trimethyl orthoacetate at 53 ° C., 53 mL (0.42 mol) of trimethyl orthoacetate And a solution of 5.0 mL (40 mmol) of n-hexanoic acid were added dropwise over 8 hours. After stirring at the same temperature for 6 hours, a solution of 5.3 mL (42 mmol) of trimethyl orthoacetate and 0.5 mL (4 mmol) of n-hexanoic acid was further added dropwise, and the mixture was further stirred at the same temperature for 2 hours. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (3: 1, 300 mL), washed with saturated aqueous sodium bicarbonate solution (twice) and saturated brine, 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 geranylgeranyl acetate) was obtained as a crude liquid. This crude product was used as it was in the next reaction.

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

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 a pressure of 200-250 mm Hg and 5,9,13,17-tetramethyl at 85 ° C. 13.9 g (40.0 mmol) of methyl octadeca-4,8,12,16-tetraenoate (methyl geranylgeranyl acetate) was gradually added dropwise and stirred at the same temperature for 6 hours. During this time, methanol generated in 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 aqueous sodium bicarbonate solution and saturated brine (twice), and then dried over magnesium sulfate. The residue obtained by filtration and concentration is purified by silica gel column chromatography (hexane / ethyl acetate = 100: 0 to 0: 100) to obtain 5.44 g (yield 33%) of the title compound as a clear liquid. Got as. The results of ¹ H-NMR measurement and viscosity measurement of the obtained compound are as follows.

¹ H-NMR spectrum (270 MHz, CDCl ₃ , TMS) δ: 1.55-1.72 (m, 15 H), 1.9-2.2 (m, 13 H), 2.27-2.45 (m , 4H), 2.53 (brs, 1 H, OH), 3.59 (dd, J = 5.4, 11.4 Hz, 1 H), 3.68 (dd, J = 3, 11.4 Hz, 1 H) , 3.92 (m, 1H), 4.15 (dd, J = 6.0, 11.6 Hz, 1 H), 4.21 (dd, J = 4.8, 11.6 Hz, 1 H), 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 referred to as geranylgeranyl acetate glyceryl.

[Example 7] Preparation of emulsion According to the compounding ratio shown in Table 1, mannitol (D (-)-mannitol, Wako Pure Chemical Industries, Ltd.) or trehalose (D (+)-trehalose dihydrate), Tokyo, as an excipient Kasei Kogyo Co., Ltd.) and Pluronic ^(R) F127 (Unirube ^(R) 70 DP-950 B, NOF Corporation, or Aldrich P2443) as a surfactant were dissolved in water (purified water or water for injection). The resulting aqueous solution was added to the lipid and then stirred with a kettle or a vortex mixer to form a suspension. As the lipid, the compound synthesized in Examples 1 to 6, GMO (glyceryl monooleate, Riken Vitamin Co., Ltd.), or phytantriol (Tokyo Kasei Kogyo Co., Ltd.) was used. Furthermore, the white emulsion (No. 1a, 2a) containing microparticles is dispersed by dispersing this suspension with an ultrasonic homogenizer (Sonics Vibra-Cell VCX-750, manufactured by Sonics & Materials, Inc., output 150 W, 2 minutes). , 8a and 9-11) were prepared (Table 1). In addition, white emulsions (No. 1b, 2b, 3 to 7, and 8b) containing fine particles are prepared by dispersing the above suspension with a high pressure homogenizer (Starbast minimo, manufactured by Sugino Machine, pressure 150 MPa). (Table 1). The emulsions were prepared in amounts of 20-25 g each.

  The above emulsion could also be prepared by dispersing with a high-speed rotary homogenizer (POLYTRON PT3100, manufactured by KINEMATICA AG, rotation speed 15,000 rpm, about 5 minutes).

Example 8 Preparation of Powdered Particulate Formulation The emulsions (Nos. 1a, 2a, and 8a, 9 to 11) prepared in Example 7 were prepared using a glass spray vial (spray vial No. 2, manufactured by Maruem). Into a stainless steel pan, it was jetted against liquid nitrogen stirred by a stirrer tip. The obtained lipid suspension in liquid nitrogen was transferred to a stainless steel container and dried for 24 hours using a vacuum lyophilizer (FZ-6, manufactured by Labconco) to obtain a powder microparticle preparation (No. 12, 14, 21, 23, 25 and 26; Table 2). In addition, the preparation method of the powder microparticle preparation by the spray lyophilization method using a spray vial is called DV method here.

  Furthermore, in place of the glass spray vial, powder fine particles are prepared using a spray drier (Spray drier SD-1000, EYELA) equipped with a two-fluid air atomizing nozzle (1/4 J series, Spraying Systems Japan Co., Ltd.) The formulation was prepared. Specifically, the emulsions prepared in Example 7 (No. 1b, 2b, 3 to 7, 8b, and 7) using the liquid transfer pump, the spray compressor, and the two-fluid air atomizing nozzle of the spray dryer. 9) was injected at a liquid transfer rate of 5 mL / min and a spray pressure of 100 kPa against liquid nitrogen stirred in a stainless steel pan with a stirrer tip. The obtained lipid suspension in liquid nitrogen was transferred to a stainless steel container and dried for 24 hours using a vacuum lyophilizer (FZ-6, manufactured by Labconco) to obtain a powder microparticle preparation (No. 13, 15-20, 22 and 24; Table 2). In addition, the preparation method of the powder microparticle preparation by the spray freeze-drying method using a two fluid nozzle is called DS method.

Prepared using a powder microparticle formulation by DV method; as a control, instead of the emulsion, an aqueous solution of mannitol or trehalose (Table 1 i.e., do not contain lipids and Pluronic ^(R) F127, aqueous solution vehicle alone) (No. 27 and 28; Table 2).

Example 9 SEM Observation of Powder Microparticle Preparation In order to confirm the particle size and shape of the microparticles contained in the powder microparticles preparation, powder microparticles preparation No. 1 prepared in Example 8 12, 14, 15, and 17 to 28 are attached to the sample stage of a scanning electron microscope (SEM S-3000N, Hitachi High-Technologies, Japan) and coated by gold evaporation, and then an observation image (300x to 3000x) is obtained. It acquired (figure 1-3).

  FIG. 1 shows a control preparation No. 1 in which mannitol was used. No. 27 (FIG. 1A), and powder microparticle preparations No. 1 and 2 prepared by the DV method using mannitol as an excipient. 12 (FIG. 1B), no. No. 21 (FIG. 1C), and no. The SEM observation image of 25 (FIG. 1D) was shown.

  FIG. 2 shows the control formulation No. 1 using trehalose. No. 28 (FIG. 2A), and powder particulate formulation No. 1 prepared by DV method using trehalose as an excipient. 14 (FIG. 2B), no. No. 23 (FIG. 2C), and no. The SEM observation image of 26 (FIG. 2D) was shown.

  FIG. 3 shows powder particulate formulation No. 1 prepared by the DS method. 15 (FIG. 3A), no. 17 (FIG. 3B), no. 18 (FIG. 3C), no. 19 (FIG. 3D), no. 20 (FIG. 3E), no. 22 (Fig. 3F), and No. The SEM observation image of 24 (FIG. 3G) was shown.

  Both the DV method and the DS method succeeded in micronizing the preparation containing the lipid and the excipient. As shown in the SEM photograph, since the particle size of the powder microparticle preparation is smaller than that of the DV method in the DS method, the particle diameter can be controlled by the spray method.

  Thus, it was confirmed that the powder microparticle preparation of the present invention can be prepared by different spray lyophilization methods including the DV method and the DS method.

  The particle size of the powder microparticle preparation prepared by the DS method is smaller than the particle size of the powder microparticle preparation by the DV method, and the voids in the particles of the powder microparticle preparation by the DS method are more frequently found than the powder microparticle preparation by the DV method The From this, it is considered that the powder microparticle preparation obtained by the DS method has better solubility.

Example 10 Analysis of Liquid Crystal Structure of Reconstituted Emulsion In order to confirm that the powder microparticle preparation of the present invention reconstitutes a non-lamellar liquid crystal structure in water, first, the powder microparticle preparation prepared in Example 8 (No 13, 15, 18 to 20, and 22) 50 mg was mixed with 0.2 mL of water and redispersed in water to prepare an emulsion by stirring with a vortex mixer. The resulting emulsion was subjected to small-angle X-ray scattering (SAXS) structural analysis using a NANO Viewer nanoscale X-ray structure evaluation apparatus (manufactured by Rigaku). Specifically, the emulsion was introduced into the capillary under atmospheric pressure, and was measured for 1 hour (the sample itself was under atmospheric pressure) in the apparatus under reduced pressure. The emulsions (Nos. 1b, 2b, and 5 to 7 and 8b) prepared in Example 7 were also subjected to small-angle X-ray scattering (SAXS) structural analysis using a NANO Viewer nanoscale x-ray structure evaluation apparatus.

  Emulsion No. The intensity distributions of SAXS obtained for 1b, 2b and 5 to 7 and 8b are shown in FIGS. The intensity distributions of SAXS obtained for emulsions reconstituted from 13, 15, 18-20, and 22 are shown in FIGS.

  In any of the scattering intensity distributions (FIGS. 4 to 7) obtained from each emulsion, at least three scattering peaks are observed, and the ratio of those peaks is a ratio 1 :: 3: 2 specific to inverse hexagonal liquid crystals. showed that. From this result, in each of the emulsion prepared in Example 7 and the emulsion reconstituted from the powder fine particle preparation prepared in Example 8, a liquid crystal emulsion (hexasome) in which fine particles of reverse hexagonal liquid crystal (H2) are dispersed in the aqueous phase It was shown to be.

  That is, it was revealed that the powder microparticle preparation of the present invention reconstitutes a non-lamellar liquid crystal structure in water as fine particles of the same liquid crystal structure as the emulsion before powderization.

Furthermore, the observation position (scattering peak position) of the first scattering peak of each emulsion was compared between before pulverization and after reconstitution from the powder microparticle preparation. The results are shown in Table 3. Powdered fine particle preparation No. 1 of the present invention containing a lipid having an isoprenoid type fatty chain In the emulsions reconstituted from 13, 15, 18 to 20, the emulsion No. 1 before pulverization was used. The difference in the observation position of the first scattering peak between 1b, 2b and 5 to 7 is ± 4 nm ⁻¹ respectively, and the size of the unit cell of the reconstituted liquid crystal is almost the same as before pulverization there were. On the other hand, powder fine particle preparation No. 1 in which GMO was used as a lipid. In the emulsion reconstituted from No. 22, the emulsion No. 1 before pulverization was used. The difference between the observed positions of the first scattering peak and 8b was 17 nm ⁻¹ , and the size of the unit cell of the reconstituted liquid crystal significantly changed before pulverization. This means that when reconstituting the powder microparticle preparation using GMO into an emulsion, a large change occurs in the liquid crystal structure that the original emulsion preparation had, and the change is a powder using GMO For example, it is considered to affect the drug retention and drug release of the microparticle preparation. Thus, it has been shown that the powder fine particle preparation using the amphiphilic compound represented by the above general formula (I) is less likely to cause a change in the liquid crystal structure by micronizing the emulsion.

[Example 11] Analysis of particle size distribution of emulsion, zeta potential Emulsion No. 1 emulsion prepared in Example 7 No. 1a, 1b, 2a, 3 to 7 and powder microparticle preparation No. 1 prepared in Example 8. 12 to 14 and 16 to 20 were each re-dispersed in water by the same method as in Example 10, the particles were prepared by dynamic light scattering method using Zetasizer Nano-ZS (manufactured by Malvern) The zeta potential, which indicates the size distribution and particle surface charge, was measured. The measurement sample was prepared by diluting each emulsion 1000 times with distilled water.

  The average particle size (nm) (Z-Average), PdI (polydispersity index), and zeta potential (mV) obtained as an average value of three measurements are shown in Table 4 for each emulsion.

  The particle size, PDI and zeta potential of the emulsion before pulverization and the emulsion obtained by redispersing the powder microparticle preparation with distilled water, good particle size, PDI, and zeta potential (correlation with dispersion stability of particles Show).

[Example 12] X-ray diffraction analysis of powder The powder crystal structure of each of 15 and 18 to 24 was evaluated using an X-ray diffractometer (Rigaku, Mini Flex II).
As shown in FIG. 21 (FIG. 8A), no. 22 (FIG. 8B), no. No. 23 (FIG. 8C), no. Measurement data of 24 (FIG. 8D) are shown. The powder of the DV formulation and the DS formulation was shown to be in an amorphous-like state when either mannitol or trehalose was used as an excipient. This contributes to the improvement of the separation ability at the time of redispersion of the powder.
Furthermore, in FIG. No. 15 (FIG. 9A), no. 18 (FIG. 9B), no. 19 (FIG. 9C), no. Measurement data of 20 (FIG. 9D) are shown. These powder microparticle preparations were also shown to be in an amorphous-like state.

Example 13 Flowability of Powder Powder flow was evaluated for 13, 15, 19, and 20. Gently fill each powder microparticle preparation (about 0.2 g fine scale) into a dry 10 mL glass measuring cylinder (scale: 0.1 mL steps) using a funnel, and carefully check the top of the powder layer as necessary. After conditioning, the sparse packing density (ρ _bulk ) was measured from the mass and bulk volume. Next, the container filled with the powder was repeatedly dropped at a constant speed from a fixed height, and the tap filling density (ρ _taped ) was measured from the mass and the bulk volume after tapping 100 times. The Hausner ratio was calculated according to the following equation.
Hausner's ratio = _{t tapped} / _{bul bulk}
The Hausner ratio obtained as an average of three measurements is shown in Table 5 for each powder microparticle preparation.

  These Hausner ratios showed that the powder microparticle preparation of the present invention had a constant flowability. Furthermore, considering that the powder microparticle preparation of the present invention is not sticky even though it itself contains a liquid lipid, the powder microparticle preparation of the present invention can be handled in the same manner as general powders, and its operation It was shown that there was no problem with sex.

  The powder microparticle preparation according to the present invention can reconstitute non-lamellar liquid crystal compositions useful for drug delivery, and can be used for various dosage forms. In the powder microparticle preparation according to the present invention, control of drug release, improvement of drug retention rate, improvement of drug stability and the like are easily possible.

Claims (18)

A powder microparticle preparation having non-lamellar liquid crystal reconstitution ability, containing an amphiphilic compound represented by the following general formula (I).

(Wherein, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, and m represents 1 or 2,

Represents a single bond or a double bond, and R represents a hydrophilic group having two or more hydroxyl groups)   R in the above formula is 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 The powder microparticle preparation according to claim 1, which represents a hydrophilic group from which any one hydroxyl group has been removed.   The powder microparticle preparation according to claim 1 or 2, wherein R in the formula represents a hydrophilic group in which one hydroxyl group is removed from glycerol. The amphiphilic compound is
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 The powder microparticle preparation according to any one of claims 1 to 3, which is selected from the group consisting of   The powder microparticle preparation according to any one of claims 1 to 4, further comprising a surfactant.   The powder microparticle preparation according to claim 5, wherein the surfactant is a nonionic surfactant.   The powder microparticle preparation according to claim 5 or 6, wherein the surfactant is polyoxyethylene (196) polyoxypropylene (67) glycol.   The powder microparticle preparation according to any one of claims 1 to 7, further comprising a water soluble carbohydrate.   The powder microparticle preparation according to claim 8, wherein the water-soluble carbohydrate is trehalose or mannitol.   The powder microparticle preparation according to any one of claims 1 to 9, further comprising a pharmacologically active substance. A method for producing a powder microparticle preparation having non-lamellar liquid crystal reconstitution ability, comprising drying and pulverizing an emulsion containing an amphiphilic compound represented by the following general formula (I) and a surfactant.

(Wherein, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, and m represents 1 or 2,

Represents a single bond or a double bond, and R represents a hydrophilic group having two or more hydroxyl groups)   The method according to claim 11, wherein the surfactant is a nonionic surfactant.   The method according to claim 11 or 12, wherein the surfactant is polyoxyethylene (196) polyoxypropylene (67) glycol.   14. The method of any one of claims 11-13, wherein the emulsion further comprises a water soluble carbohydrate.   15. The method of claim 14, wherein the water soluble carbohydrate is trehalose or mannitol.   The method according to any one of claims 11 to 15, wherein the emulsion further comprises a pharmacologically active substance.   The method according to any one of claims 11 to 16, comprising preparing the emulsion by homogenizing an aqueous medium containing the amphiphilic compound and a surfactant by ultrasonic dispersion method or high pressure dispersion method. Method described.   The method according to any one of claims 11 to 17, wherein the emulsion is dried and powdered by spray lyophilization.

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