Non-lamellar liquid crystal-forming composition and use thereof

Abstract

The present invention provides a novel non-lamellar liquid crystal-forming composition.　According to an aspect of the present invention, a non-lamellar liquid crystal-forming composition comprises a fatty acid ester and a phospholipid, wherein the fatty acid ester has a hydrophilic group having one hydroxyl group or a salt thereof, and does not have any of structures represented by general formulae (A)-(D). (In the formulae, each m is independently 1 or 2, a, b, c, and d each independently denote a bonding site to a hydrophilic group, n is 0, 1, or 2, and the wavy lines each independently denote an E-isomer or a Z-isomer.)

Description

Non-lamellar liquid crystal forming composition and its use

[0001] The present invention relates to a non-lamellar liquid crystal forming composition and its use.

[0002] Lyotropic liquid crystals such as liposomes have been widely reported 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 higher drug loading and ease of preparation.

[0003] Various liquid crystal-forming compounds are used for various purposes in the fields of cosmetics, pharmaceuticals, etc. For example, a sustained-release lipid pre-concentrate is known that contains a sorbitan unsaturated fatty acid ester having two or more —OH groups in the polar head group, a phospholipid, and a specific liquid crystal hardener, and that exists as a lipid liquid phase in the absence of an aqueous fluid and forms liquid crystals in the presence of an aqueous fluid (see, for example, Patent Document 1).

[0004] Another known example is a pre-formulation containing a low-viscosity non-liquid crystalline mixture of an ester of a sugar or sugar derivative, a phospholipid, and a biocompatible, oxygen-containing, low-viscosity organic solvent, which forms or is capable of forming a non-lamellar liquid crystalline phase structure when contacted with an aqueous fluid, and which does not further contain a specific liquid crystalline curing agent (see, for example, Patent Document 2).

[0005] Special table No. 2014-527545 Publication No. 2018-502091

[0006] An object of the present invention is to provide a novel non-lamellar liquid crystal forming composition. Another object of the present invention is to provide a novel compound or a salt thereof that can provide a non-lamellar liquid crystal forming composition.

[0007] The present inventors conducted extensive research to solve the above problems and discovered a non-lamellar liquid crystal-forming composition containing a specific fatty acid ester and a phospholipid. Furthermore, the present inventors succeeded in synthesizing a new compound that can be used to prepare the non-lamellar liquid crystal-forming composition. Thus, the present inventors have completed the present invention.

[0008] That is, the present invention includes the following: [1] A non-lamellar liquid crystal forming composition comprising a fatty acid ester and a phospholipid, wherein the fatty acid ester has a hydrophilic group having one hydroxyl group or a salt thereof, and the fatty acid ester is a fatty acid ester having none of the structures represented by the following general formulas (A) to (D): (In general formulas (A) to (D), m each independently represents 1 or 2, and a, b, c, and d each independently represent a bonding site with a hydrophilic group, each independently represent a single bond or a double bond, n in general formula (A) is 0, 1 or 2, and the wavy lines in general formulas (B), (C) and (D) each independently represent an E-form or a Z-form.) [2] The composition according to [1], wherein the number of carbon atoms in the fatty acid of the fatty acid ester is 6 to 24. [3] The composition according to [1] or [2], wherein the weight ratio of the fatty acid ester to the phospholipid is 90:10 to 20:80. [4] The composition according to any one of [1] to [3], wherein the fatty acid ester is a fatty acid ester of at least one selected from saturated or unsaturated straight-chain fatty acids and derivatives thereof and at least one selected from glycols and derivatives thereof.

[0009] [5] The composition according to [4], wherein the straight-chain fatty acid has an unsaturation degree of 0 to 6. [6] The composition according to [4] or [5], wherein the straight-chain fatty acid is a straight-chain fatty acid represented by the following general formula (I): (In the general formula (I), R is —C ｐ H ｑq is an integer from 2p-4 to 2p when p is 5 or less, an integer from 2p-6 to 2p when p is 6 or 7, an integer from 2p-8 to 2p when p is 8 or 9, an integer from 2p-10 to 2p when p is 10 or 11, and an integer from 2p-12 to 2p when p is 12 or more.) [7] The composition according to any one of [4] to [6], wherein the linear fatty acid is at least one fatty acid selected from caprylic acid, lauric acid, myristic acid, palmitic acid, oleic acid, linoleic acid, and behenic acid. [8] The composition according to any one of [4] to [7], wherein the glycol is at least one glycol selected from propylene glycol, ethylene glycol, butylene glycol, 3-methyl-1,3-butanediol, diethylene glycol, and isosorbide. [9] The composition according to any one of [1] to [8], wherein the phospholipid is at least one phospholipid selected from phosphatidylcholine and phosphatidylethanolamine, and salts thereof.

[10] The composition according to any one of [1] to [9], wherein the phospholipid is at least one phospholipid selected from soybean phosphatidylcholine, egg yolk phosphatidylcholine, dimyristoylphosphatidylcholine, dioleylphosphatidylcholine, and dioleylphosphatidylethanolamine, and salts thereof.

[11] The composition according to any one of [1] to

[10] , comprising at least one of an oil and an organic solvent.

[12] The composition according to any one of [1] to

[11] , which is a non-lamellar liquid crystal composition further comprising an aqueous medium.

[13] The composition according to

[12] , which is a non-lamellar liquid crystal emulsion composition further comprising a surfactant.

[14] The composition according to any one of [1] to

[11] , which is a liquid crystal precursor composition capable of forming non-lamellar liquid crystals in the presence of an aqueous medium.

[0010]

[15] A pharmaceutical preparation comprising the composition according to any one of [1] to

[14] .

[16] The pharmaceutical preparation according to

[15] , for preventing adhesion of biological tissues.

[17] The pharmaceutical preparation according to

[15] , wherein the composition is a sustained-release preparation further comprising a drug.

[18] The pharmaceutical preparation according to

[17] , wherein the drug is a gonadotropin-releasing hormone (GnRH) agonist.

[19] The pharmaceutical preparation according to

[18] , wherein the GnRH agonist is leuprolide or a salt thereof.

[20] The pharmaceutical preparation according to any one of

[15] to

[19] , which is in the form of a spray, aerosol, injection, or depot preparation.

[21] A compound represented by formula (X') or a salt thereof. (In the general formula (X′), R １ is a structure derived from propylene glycol, butylene glycol, isoprene glycol, diethylene glycol or isosorbide.)

[22] A compound represented by formula (X), (Y), or (Z) or a salt thereof. This specification includes the disclosure of Japanese Patent Application No. 2021-122489, from which this application claims priority.

[0011] The present invention can provide a new composition capable of forming a non-lamellar liquid crystal.

[0012] Figure 1 shows the release rate (in vitro release data) of the drug (leuprolide acetate) from precursor formulations No. 32, No. 33, and No. 35 formulated with leuprolide acetate in Example 5. Figure 2 shows the drug (leuprolide acetate) kinetic data from precursor formulations No. 34 and No. 35 formulated with leuprolide acetate in Example 6. Figure 3 shows a photograph of the administration site of a rat to which precursor formulation No. 35 in Example 6 was subcutaneously administered.

[0013] The present invention will be described in detail below.

[0014] One aspect of the present invention relates to a non-lamellar liquid crystal forming composition comprising a fatty acid ester and a phospholipid, wherein the fatty acid ester has a hydrophilic group having one hydroxyl group or a salt thereof, and the fatty acid ester is a fatty acid ester that does not have any of the structures represented by the following general formulas (A) to (D): The non-lamellar liquid crystal forming composition according to the present invention has excellent biocompatibility due to the inclusion of a phospholipid, and such a non-lamellar liquid crystal forming composition is highly safe and therefore acceptable for in vivo application.

[0015] The fatty acid ester used in the present invention has a hydrophilic group having one hydroxyl group or a salt thereof. The fatty acid ester used in the present invention is a fatty acid ester that does not have any of the structures represented by the following general formulas (A) to (D). A fatty acid ester having a hydrophilic group is also referred to as an amphiphilic compound. The fatty acid ester may be used alone or in combination of two or more.

[0016]

[0017] (In general formulas (A) to (D), m each independently represents 1 or 2, and a, b, c, and d each independently represent a bonding site with a hydrophilic group, each independently represents a single bond or a double bond, n in general formula (A) is 0, 1 or 2, and the wavy lines in general formulas (B), (C) and (D) each independently represent an E-form or a Z-form.

[0018] The structures represented by the general formulas (A) to (D) are so-called isoprenoid fatty chains. One of the characteristics of the fatty acid ester used in the present invention is that it does not have a so-called isoprenoid fatty chain. The bonding site with the hydrophilic group means, in other words, a bonding hand with the hydrophilic group.

[0019] The fatty acid ester has a hydrophilic group having one hydroxyl group or a salt thereof. The hydrophilic group is usually a hydrophilic group composed of a carbonyl (C═O) structure derived from the fatty acid that constitutes the fatty acid ester and a structure derived from a compound (e.g., glycol) that forms an ester bond with the fatty acid. The hydrophilic group has one hydroxyl group or a salt thereof, and examples of the salt of the hydroxyl group include metal salts of the hydroxyl group (e.g., alkali metal salts, alkaline earth metal salts, transition metal salts). Preferred examples of the salt of the hydroxyl group include alkali metal salts of the hydroxyl group and alkaline earth metal salts of the hydroxyl group, and particularly preferred examples include sodium salts of the hydroxyl group (—ONa), potassium salts of the hydroxyl group (—OK), and calcium salts of the hydroxyl group (—OCa). １／２ ), magnesium salt of hydroxyl group (-OMg １／２ ) are examples. The salt of the hydroxyl group may be any other pharmaceutically acceptable salt. The present inventors have found that using a fatty acid ester having a hydrophilic group with one hydroxyl group or a salt thereof as the fatty acid ester can reduce the viscosity of the composition compared to using a fatty acid ester in which the hydrophilic group has multiple hydroxyl groups or salts thereof. A composition with low viscosity is useful, for example, when used as an injection, because it allows for the use of a thinner injection needle. Furthermore, compositions with low viscosity tend to be easier to prepare and more stable over a wide temperature range than compositions with high viscosity. Furthermore, compositions containing fatty acid esters in a wide range of weight ratios are preferred because they can be formed into liquid compositions.

[0020] The number of carbon atoms of the fatty acid of the fatty acid ester, in other words, the number of carbon atoms of the fatty acid constituting the fatty acid ester (the fatty acid that is the raw material for the fatty acid ester), is preferably 6 to 24, more preferably 8 to 22, and even more preferably 8 to 18. A carbon atom number within the above range is preferred because a non-lamellar liquid crystal-forming composition can be easily obtained. The fatty acid may be a branched fatty acid. As the branched fatty acid, saturated or unsaturated branched fatty acids can be used. Examples of branched fatty acids include 2-ethylhexanoic acid, isononanoic acid, isodecanoic acid, isotridecanoic acid, isomyristic acid, isopalmitic acid, isostearic acid, and isoeicosanoic acid.

[0021] In one preferred embodiment, the fatty acid is a saturated or unsaturated straight-chain fatty acid. The fatty acid may be a synthetic fatty acid, a semi-synthetic fatty acid, or a natural fatty acid, but a natural fatty acid is one preferred embodiment. In another preferred embodiment, the fatty acid ester is a fatty acid ester of at least one selected from fatty acids and their derivatives and at least one selected from glycols and their derivatives.

[0022] That is, one preferred embodiment is a fatty acid ester of at least one selected from saturated or unsaturated straight-chain fatty acids and their derivatives and at least one selected from glycols and their derivatives. There are no particular limitations on the method for producing the fatty acid ester, and for example, it can be obtained by reacting at least one selected from saturated or unsaturated straight-chain fatty acids and their derivatives with at least one selected from glycols and their derivatives under known conditions, reacting the carboxyl group derived from the fatty acid with the hydroxyl group derived from the glycol to form an ester bond. As a specific example, the fatty acid ester can be produced by the method described in the Examples below.

[0023] When the straight-chain fatty acid is an unsaturated fatty acid, its degree of unsaturation, i.e., the number of carbon-carbon double bonds, is usually 1 or more, preferably 1 to 6, more preferably 1 to 4, and particularly preferably 1 or 2. In other words, the straight-chain fatty acid is preferably a straight-chain fatty acid having a degree of unsaturation of 0 to 6, more preferably a straight-chain fatty acid having a degree of unsaturation of 0 to 4, and particularly preferably a straight-chain fatty acid having a degree of unsaturation of 0 to 2. Furthermore, when the straight-chain fatty acid is an unsaturated fatty acid having a degree of unsaturation of 1 or 2, a preferred embodiment from the viewpoint of the stability of the composition is one in which the number of carbon atoms is 18 or less, and when the straight-chain fatty acid is a saturated fatty acid, a preferred embodiment from the viewpoint of the stability of the composition is one in which the number of carbon atoms is 16 or less.

[0024] In one preferred embodiment, the straight-chain fatty acid is a straight-chain fatty acid represented by the following general formula (I): That is, in one preferred embodiment, the straight-chain fatty acid has a methyl group at one end.

[0025] (In the general formula (I), R is —C ｐ H ｑ -, p is an integer of 4 to 22, q is an integer of 2p-4 to 2p when p is 5 or less, an integer of 2p-6 to 2p when p is 6 or 7, an integer of 2p-8 to 2p when p is 8 or 9, an integer of 2p-10 to 2p when p is 10 or 11, and an integer of 2p-12 to 2p when p is 12 or more.

[0026] p is more preferably an integer of 6 to 20, and even more preferably an integer of 6 to 16. In one preferred embodiment, q is 2p-4 to 2p, regardless of the value of p. When q is 2p-12, -C ｐ H ｑ The linear hydrocarbon group represented by - has a degree of unsaturation of 6 and when q is 2p-10, it is represented by -C ｐ H ｑ The linear hydrocarbon group represented by - has a degree of unsaturation of 5 and when q is 2p-8, it is -C ｐ H ｑ The linear hydrocarbon group represented by - has an unsaturation degree of 4 and when q is 2p-6, it is -C ｐ H ｑ The linear hydrocarbon group represented by - has a degree of unsaturation of 3 and when q is 2p-4, it is represented by -C ｐ H ｑ The linear hydrocarbon group represented by - has a degree of unsaturation of 2 and when q is 2p-2, it is represented by -C ｐ H ｑ The linear hydrocarbon group represented by - has a degree of unsaturation of 1 and when q is 2p, it is -C ｐ H ｑThe linear hydrocarbon group represented by - is a saturated linear hydrocarbon group (linear alkylene group, degree of unsaturation: 0). In addition, the above-mentioned 2p-4 to 2p mean 2p-4, 2p-2, or 2p, 2p-6 to 2p mean 2p-6, 2p-4, 2p-2, or 2p, 2p-8 to 2p mean 2p-8, 2p-6, 2p-4, 2p-2, or 2p, 2p-10 to 2p mean 2p-10, 2p-8, 2p-6, 2p-4, 2p-2, or 2p, and 2p-12 to 2p mean 2p-12, 2p-10, 2p-8, 2p-6, 2p-4, 2p-2, or 2p.

[0027] Examples of the fatty acids include butanoic acid, pentanoic acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, lauric acid, myristic acid, pentadecylic acid, palmitic acid, palmitoleic acid, margaric acid, stearic acid, oleic acid, vaccenic acid, linoleic acid, (9,12,15)-linolenic acid, (6,9,12)-linolenic acid, eleostearic acid, arachidic acid, 8,11-eicosadienoic acid, mead acid, arachidonic acid, behenic acid, eicosapentaenoic acid, docosahexaenoic acid, lignoceric acid, and nervonic acid. The fatty acids may be used alone or in combination. Straight-chain fatty acids are preferred, and at least one fatty acid selected from caprylic acid, lauric acid, myristic acid, palmitic acid, oleic acid, linoleic acid, and behenic acid is particularly preferred. The fatty acid may be used alone or in combination of two or more. Note that commercially available fatty acids may be mixtures of multiple fatty acids, and such fatty acids may also be used without any problems.

[0028] The glycol refers to a compound consisting of a chain or ring of carbon, oxygen, and hydrogen, in which two hydroxyl groups are bonded to two different carbon atoms. The glycol is preferably at least one glycol selected from propylene glycol, ethylene glycol, butylene glycol, 3-methyl-1,3-butanediol, diethylene glycol, and isosorbide.

[0029] When the two hydroxyl groups possessed by a glycol are unequal, for example, when one of the hydroxyl groups possessed by a glycol is a primary hydroxyl group and the other is a secondary hydroxyl group, the fatty acid ester will have isomers with different hydroxyl groups bonded to the fatty acid. The fatty acid ester used in the present invention may be one of the isomers or a mixture containing both isomers. Generally, when a glycol has a primary hydroxyl group and a secondary hydroxyl group, it is believed that the fatty acid ester synthesized will contain a higher proportion of fatty acid esters in which the primary hydroxyl group is bonded to the fatty acid than fatty acid esters in which the secondary hydroxyl group is bonded to the fatty acid. In this example, the chemical formula of the synthesized fatty acid ester is shown, but this merely shows a representative structure and does not deny the existence of isomers in which different hydroxyl groups possessed by a glycol are bonded to a fatty acid.

[0030] The two hydroxyl groups of the glycol are unequal, for example, when propylene glycol, 1,3-butylene glycol, 3-methyl-1,3-butanediol, isosorbide, or the like is used as the glycol.

[0031] Examples of derivatives of saturated or unsaturated straight-chain fatty acids include alkyl esters of the aforementioned saturated or unsaturated straight-chain fatty acids (esters of saturated or unsaturated straight-chain fatty acids and monoalcohols), and salts of saturated or unsaturated straight-chain fatty acids (e.g., metal salts). Examples of glycol derivatives include ethers of the aforementioned glycols and monools (e.g., monoethers of glycols in which one of the hydroxyl groups of glycol is etherified with a monool), and salts of glycols (e.g., metal salts). The saturated or unsaturated straight-chain fatty acid derivatives may be used alone or in combination of two or more. Commercially available saturated or unsaturated straight-chain fatty acid derivatives may be mixtures of multiple components (mixtures of multiple types of saturated or unsaturated straight-chain fatty acid derivatives), but such saturated or unsaturated straight-chain fatty acid derivatives can also be used without any problems.

[0032] Preferable examples of fatty acid esters used in the present invention include propylene glycol monocaprylate, propylene glycol monooleate, propylene glycol monomyristate, propylene glycol monopalmitate, propylene glycol monolinoleate, propylene glycol monobehenate, ethylene glycol monocaprylate, ethylene glycol monooleate, ethylene glycol monomyristate, ethylene glycol monopalmitate, ethylene glycol monolinoleate, ethylene glycol monobehenate, 1,3-butylene glycol monocaprylate, 1,3-butylene glycol monooleate, 1,3-butylene glycol monomyristate, 1,3-butylene glycol monopalmitate, 1,3-butylene glycol monolinoleate, 1,3-butylene glycol monobehenate, 3-methyl-1,3-butanediol monocaprylate, Examples of the fatty acid ester include 3-methyl-1,3-butanediol monooleate, 3-methyl-1,3-butanediol monomyristate, 3-methyl-1,3-butanediol monopalmitate, 3-methyl-1,3-butanediol monolinoleate, 3-methyl-1,3-butanediol monobehenate, diethylene glycol monocaprylate, diethylene glycol monooleate, diethylene glycol monomyristate, diethylene glycol monopalmitate, diethylene glycol monolinoleate, diethylene glycol monobehenate, isosorbide monocaprylate, isosorbide monooleate, isosorbide monomyristate, isosorbide monopalmitate, isosorbide monolinoleate, and isosorbide monobehenate. Other examples include, but are not limited to, salts of hydroxyl groups in the hydrophilic groups of the aforementioned fatty acid esters. The salt of the hydroxyl group is preferably a sodium salt of the hydroxyl group (-ONa), a potassium salt of the hydroxyl group (-OK), or a calcium salt of the hydroxyl group (-OCa １／２ ), magnesium salt of hydroxyl group (-OMg １／２ ), and other pharmaceutically acceptable salts.

[0033] Particularly preferred examples of the fatty acid ester used in the present invention include propylene glycol monooleate, propylene glycol monolinoleate, ethylene glycol monooleate, ethylene glycol monolinoleate, 1,3-butylene glycol monooleate, 1,3-butylene glycol monolinoleate, 3-methyl-1,3-butanediol monooleate, 3-methyl-1,3-butanediol monolinoleate, diethylene glycol monooleate, diethylene glycol monolinoleate, isosorbide monooleate, and isosorbide monolinoleate. Other examples include, but are not limited to, salts of the hydroxyl groups of the hydrophilic groups of the aforementioned fatty acid esters. Preferred examples of the salts of the hydroxyl groups include sodium salts of the hydroxyl groups (-ONa), potassium salts of the hydroxyl groups (-OK), and calcium salts of the hydroxyl groups (-OCa). １／２ ), magnesium salt of hydroxyl group (-OMg １／２ ), and other pharmaceutically acceptable salts.

[0034] The non-lamellar liquid crystal forming composition according to the present invention may contain one or more of the above-mentioned fatty acid esters. When the non-lamellar liquid crystal forming composition according to the present invention contains a plurality of the above-mentioned fatty acid esters, the weight ratio of these fatty acid esters is not particularly limited.

[0035] Among the fatty acid esters used in the present invention, the compound represented by formula (X') is a novel compound, and one example of this embodiment is a compound represented by formula (X') or a salt thereof. (In the general formula (X′), R １ is a structure derived from propylene glycol, butylene glycol, isoprene glycol, diethylene glycol, or isosorbide.

[0036] The compound represented by formula (X') is a fatty acid ester of linoleic acid and propylene glycol, butylene glycol, isoprene glycol (3-methyl-1,3-butanediol), diethylene glycol, or isosorbide. That is, the structure derived from propylene glycol, butylene glycol, isoprene glycol, diethylene glycol, or isosorbide means a structure in which a hydrogen atom constituting one hydroxyl group is removed from propylene glycol, butylene glycol, isoprene glycol, diethylene glycol, or isosorbide. As an example, R １ is a structure derived from diethylene glycol, the structure is —O—CH ２ -CH ２ -O-CH ２ -CH ２ It is —OH.

[0037] Among the fatty acid esters used in the present invention, the compounds represented by formula (X), (Y), or (Z) are novel compounds, and one example of the present embodiment includes a compound represented by formula (X), (Y), or (Z) or a salt thereof.

[0038]

[0039] The compound represented by formula (X) is propylene glycol monolinoleate (propylene glycol monolinoleate (C18:2)), the compound represented by formula (Y) is 1,3-butylene glycol monooleate (butylene glycol monooleate (C18:1)), and the compound represented by formula (Z) is isosorbide monooleate.

[0040] Salts of the compounds represented by formula (X'), (X), (Y), or (Z) include salts of hydroxyl groups in the molecule, specifically alkali metal salts of hydroxyl groups, and alkaline earth metal salts of hydroxyl groups. Particularly preferred examples include sodium salts of hydroxyl groups (-ONa), potassium salts of hydroxyl groups (-OK), and calcium salts of hydroxyl groups (-OCa). １／２ ), magnesium salt of hydroxyl group (-OMg １／２ The salt of the hydroxyl group may be any other pharmaceutically acceptable salt.

[0041] In the present invention, the aforementioned fatty acid ester can be used in combination with a phospholipid to produce a non-lamellar liquid crystal-forming composition. That is, the present invention provides a composition, particularly a non-lamellar liquid crystal-forming composition, comprising the aforementioned fatty acid ester and a phospholipid.

[0042] The aforementioned fatty acid esters can be used in combination with phospholipids. With some exceptions, the aforementioned fatty acid esters alone were unable to form nonlamellar liquid crystals. However, after extensive research, the inventors discovered that nonlamellar liquid crystals can be formed by combining the aforementioned fatty acid esters with phospholipids. Furthermore, if the aforementioned fatty acid esters are toxic, the phospholipids can reduce the toxicity of the aforementioned fatty acid esters, improving the biocompatibility of the nonlamellar liquid crystal-forming composition and increasing safety when applied to the body. In the present invention, "biocompatibility" refers to the property of causing little or no adverse reactions (side effects) in the body when applied to the body. "Toxicity" in the present invention includes, but is not limited to, systemic toxicity such as hepatotoxicity, foreign body reactions such as abscess formation, and local toxicity resulting in tissue damage such as bleeding and discoloration. Hepatotoxicity can be confirmed by the occurrence of at least one symptom indicative of liver damage, such as the occurrence of ascites, liver enlargement, adhesions around the liver (especially adhesions in undamaged and uninflamed areas), and liver pallor. The non-lamellar liquid crystal forming composition according to the present invention, which comprises a fatty acid ester and a phospholipid, is acceptable for internal administration (preferably parenteral administration such as intraperitoneal administration, intramuscular administration, or subcutaneous administration). In the present invention, "acceptable for internal administration" means that when administered to a living body (typically, when administered parenterally into the body such as intraperitoneal administration, intramuscular administration, or subcutaneous administration), no toxicity occurs or the toxicity occurs at a pharmaceutically acceptable level. However, the non-lamellar liquid crystal forming composition according to the present invention is not limited to compositions intended for internal administration.

[0043] The phospholipids used in the present invention include, but are not limited to, one or more phospholipids selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerin, phosphatidic acid, and sphingomyelin, and salts thereof, or phospholipid preparations or fractions containing the same. Examples of phosphatidylcholine include, but are not limited to, dioleylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), soybean phosphatidylcholine (SPC; also known as soybean lecithin), and egg yolk phosphatidylcholine (EPC; also known as egg yolk lecithin). Examples of phosphatidylethanolamine include, but are not limited to, dioleylphosphatidylethanolamine (DOPE). An example of a phosphatidylglycerin is dioleylphosphatidylglycerin, and an example of a salt of phosphatidylglycerin is sodium dioleylphosphatidylglycerin (DOPG-Na), but these are not limited thereto. The phospholipids used in the present invention may be synthetic or naturally derived. In one embodiment, the phospholipid used in the present invention may be a phosphatidylcholine, such as soybean phosphatidylcholine or egg yolk phosphatidylcholine. In another embodiment, the phospholipid used in the present invention may be a phosphatidylcholine, phosphatidylethanolamine, or phosphatidylglycerin, or a salt thereof. In one embodiment, the phospholipid used in the present invention may be selected from the group consisting of soybean phosphatidylcholine (SPC), egg yolk phosphatidylcholine (EPC), dimyristoylphosphatidylcholine (DMPC), dioleylphosphatidylcholine (DOPC), and dioleylphosphatidylethanolamine (DOPE). In a preferred embodiment, the phospholipid may be at least one phospholipid selected from phosphatidylcholine and phosphatidylethanolamine, and salts thereof.In another preferred embodiment, the phospholipid may be at least one phospholipid selected from soybean phosphatidylcholine, egg yolk phosphatidylcholine, dimyristoylphosphatidylcholine, dioleylphosphatidylcholine, and dioleylphosphatidylethanolamine, and salts thereof. The non-lamellar liquid crystal forming composition according to the present invention may contain one or more phospholipids.

[0044] For the purpose of improving the biocompatibility of the amphiphilic compound and increasing the safety of the non-lamellar liquid crystal forming composition, the weight ratio of the fatty acid ester to the phospholipid contained in the non-lamellar liquid crystal forming composition according to the present invention is, but is not limited to, preferably 90:10 to 10:90, more preferably 90:10 to 20:80, and particularly preferably 80:20 to 20:80. Other examples of the ratio include fatty acid ester:phospholipid = 80:20 to 30:70, 70:30 to 10:90, 70:30 to 20:80, 70:30 to 30:70, 60:40 to 10:90, 60:40 to 20:80, 60:40 to 30:70, 60:40 to 40:60, 45:55 to 10:90, and 45:55. The ratio may be 50:50 to 20:80, 45:55 to 30:70, 45:55 to 35:65, 45:55 to 55:45, 50:50 to 20:80, 50:50 to 30:70, 40:60 to 10:90, 40:60 to 20:80, 40:60 to 30:70, 35:65 to 20:80, or 35:65 to 25:75.

[0045] The weight ratio of the fatty acid ester to the phospholipid is calculated using the total weight of the fatty acid esters when multiple types of fatty acid esters are used, and the total weight of the phospholipids when multiple types of phospholipids are used. In this specification, the terms "weight" and "mass" are used interchangeably.

[0046] In an alternative embodiment, the non-lamellar liquid crystal-forming composition of the present invention may not contain a phospholipid, as long as it has sufficiently high biocompatibility (i.e., safety) and non-lamellar liquid crystal-forming properties, depending on the application, dosage form, composition, etc. Therefore, the present invention also relates to a composition, particularly a non-lamellar liquid crystal-forming composition, containing the fatty acid ester. Note that in the alternative embodiment, fatty acid esters having a hydrophilic group with one hydroxyl group or a salt thereof, derived from glycols without an ether bond, tend to be difficult to form non-lamellar liquid crystals without the use of phospholipids and may not be usable. On the other hand, fatty acid esters having a hydrophilic group with one hydroxyl group or a salt thereof, derived from glycols with an ether bond, such as diethylene glycol monooleate, tend to be suitable for use in the alternative embodiment. Such non-lamellar liquid crystal-forming compositions can also be used as sustained-release formulations, for example, when containing a drug.

[0047] In the present invention, the term "non-lamellar liquid crystal forming composition" means a composition that forms a non-lamellar liquid crystal structure (non-lamellar liquid crystal composition), or a composition that does not form a non-lamellar liquid crystal structure by itself but has the ability to form a non-lamellar liquid crystal structure in the presence of water (i.e., upon contact with an aqueous medium) (liquid crystal precursor composition).

[0048] When the non-lamellar liquid crystal-forming composition according to the present invention is a liquid crystal precursor composition, it does not contain an aqueous medium or does not contain an amount of aqueous medium sufficient to form a non-lamellar liquid crystal structure. That is, in one embodiment, the non-lamellar liquid crystal-forming composition according to the present invention is a liquid crystal precursor composition capable of forming non-lamellar liquid crystals in the presence of an aqueous medium. In a more specific embodiment, the non-lamellar liquid crystal-forming composition according to the present invention is a liquid crystal precursor composition that does not contain an aqueous medium and is capable of forming non-lamellar liquid crystals in the presence of an aqueous medium. When the non-lamellar liquid crystal-forming composition according to the present invention is a non-lamellar liquid crystal composition, it contains an aqueous medium, preferably an amount of aqueous medium sufficient to form a non-lamellar liquid crystal structure. The aqueous medium is not particularly limited and may be sterilized water, purified water, distilled water, ion-exchanged water, ultrapure water, water for injection, physiological saline, phosphate buffer solution, or the like. The non-lamellar liquid crystal-forming composition according to the present invention may be a liquid crystal emulsion (more specifically, a non-lamellar liquid crystal emulsion composition). The non-lamellar liquid crystal-forming composition according to the present invention, which is a liquid crystal emulsion, preferably further contains a surfactant. A non-lamellar liquid crystal emulsion composition is also referred to as a dispersant. The non-lamellar liquid crystal emulsion composition according to the present invention, which contains the above-mentioned amphiphilic compound and phospholipid, also exhibits high stability.

[0049] An example of a surfactant used in the non-lamellar liquid crystal forming composition according to the present invention is P80 (polyoxyethylene sorbitan monooleate (20E.O.)). Other examples of surfactants 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. Nonionic surfactants having a molecular weight of 1,000 or more (more preferably 5,000 or more) are 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 commercially available under various names, such as Pluronic (registered trademark), Poloxamer (registered trademark), Unilube (registered trademark), and Pronon (registered trademark). Particularly preferred examples of nonionic surfactants include polyoxyethylene (200) polyoxypropylene (70) glycol and polyoxyethylene (196) polyoxypropylene (67) glycol (also known as Pluronic (registered trademark) F127; Unilube 70DP-950B, Poloxamer (registered trademark) 407). In the present invention, the fatty acid ester acts as an amphiphilic compound due to the presence of a hydrophilic group, but is not included in the scope of the surfactant. The non-lamellar liquid crystal forming composition according to the present invention may contain one or more of such surfactants.

[0050] The non-lamellar liquid crystal forming composition according to the present invention may contain at least one of an oil component and an organic solvent.

[0051] Examples of oils that can be used in the non-lamellar liquid crystal forming composition of the present invention include, but are not limited to, vegetable oils such as sesame oil, soybean oil, corn oil, coconut oil, safflower oil, perilla oil, olive oil, castor oil, and cottonseed oil, animal oils such as egg yolk oil, fish oil, and lanolin, medium-chain triglycerides (MCT), triglycerides, mineral oils such as liquid paraffin, hydrocarbon oils such as squalene and squalane, ester oils such as isopropyl myristate (IPM), cholesterol, tocopherol, tocopherol acetate, glyceryl dioleate (GDO), gelling hydrocarbons, tetrahydrofarnesyl methyl acetate, hexahydrogeranylgeranyl methyl acetate, etc. Preferably, the oil is pharmaceutically acceptable.

[0052] The total amount of the fatty acid ester, phospholipid, and oil contained in the non-lamellar liquid crystal-forming composition according to the present invention is not particularly limited. When the non-lamellar liquid crystal-forming composition according to the present invention is a liquid crystal precursor composition, the total amount of the fatty acid ester, phospholipid, and oil may be, for example, 30% or more of the total amount of the composition, typically 60 to 100%, preferably 65 to 95%, for example, 75 to 95%, 75 to 93%, or 80 to 95%. When the non-lamellar liquid crystal-forming composition according to the present invention is a liquid crystal emulsion, the total amount of the fatty acid ester, phospholipid, and oil may be, for example, 0.01 to 40%, preferably about 1 to 30%, for example, 20 to 30%, 20 to 23%, or 25 to 30%, depending on the application of the liquid crystal emulsion.

[0053] In this specification, the percentage (%) of a component in the non-lamellar liquid crystal forming composition means % by weight and can be expressed in units of w / w%.

[0054] Examples of organic solvents used in the non-lamellar liquid crystal forming composition of the present invention include, but are not limited to, alcohols such as ethanol, propylene glycol, and isopropanol; ethers such as diethyl ether and polyethylene glycol; dimethyl sulfoxide (DMSO), N-methylpyrrolidone (NMP); and dimethylacetamide (DMA). The organic solvent is preferably pharmaceutically acceptable. In the non-lamellar liquid crystal forming composition of the present invention, a protic organic solvent or an aprotic organic solvent may be used alone, or a protic organic solvent and an aprotic organic solvent may be used in combination. Examples of protic organic solvents include alcohols such as ethanol and propylene glycol, and polyethylene glycol. Examples of aprotic organic solvents include ethers such as diethyl ether, dimethyl sulfoxide (DMSO), N-methylpyrrolidone (NMP), and dimethylacetamide (DMA).

[0055] When the non-lamellar liquid crystal forming composition according to the present invention contains an oil, the ratio of the total weight of the fatty acid ester and the phospholipid contained in the non-lamellar liquid crystal forming composition to the weight of the oil is preferably 30:70 to 99:1, more preferably 40:60 to 98:2, and particularly preferably 45:55 to 97:3. Furthermore, when the non-lamellar liquid crystal forming composition according to the present invention contains an organic solvent, the ratio of the total weight of the fatty acid ester, the phospholipid, and any oil contained in the non-lamellar liquid crystal forming composition to the weight of the organic solvent is preferably 80:20 to 99:1, more preferably 85:15 to 97:3, and particularly preferably 90:10 to 95:5.

[0056] The non-lamellar liquid crystal forming composition according to the present invention may contain a water-soluble polymer, such as, but not limited to, hydroxypropyl cellulose (HPC), hydroxyethyl cellulose, polyvinylpyrrolidone, carbopol, carrageenan, chitosan, chondroitinate, xanthan gum, hyaluronate (such as sodium hyaluronate), alginate (such as sodium alginate), gelatin, and dextran. Examples of hydroxypropyl cellulose (HPC) include five grades of HPC commercially available from Nippon Soda Co., Ltd. (Japan): HPC-SSL (molecular weight: about 40,000, viscosity: 2 to 2.9 mPa·s), HPC-SL (molecular weight: about 100,000, viscosity: 3 to 5.9 mPa·s), HPC-L (molecular weight: about 140,000, viscosity: 6 to 10 mPa·s), HPC-M (molecular weight: about 620,000, viscosity: 150 to 400 mPa·s), and HPC-H (molecular weight: about 910,000, viscosity: 1000 to 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 to 700,000 or 10,000 to 80,000.

[0057] The non-lamellar liquid crystal forming composition of the present invention may contain an antioxidant, such as ascorbic acid or sodium sulfite, but is not limited to these.

[0058] The non-lamellar liquid crystal forming composition of the present invention can form a gel containing water or in the presence of water. This gel formation means that non-lamellar liquid crystals (liquid crystal gels) are formed. The non-lamellar liquid crystals can retain and sustainably release substances such as drugs. The present invention also relates to a gel (gel-like composition) containing the non-lamellar liquid crystal forming composition of the present invention.

[0059] The non-lamellar liquid crystal is a liquid crystal structure that is not a lamellar liquid crystal, and specifically may be, for example, a cubic liquid crystal, a reversed hexagonal liquid crystal (HII), a reversed micellar cubic phase (Fd3m), or a sponge phase (L3). The non-lamellar liquid crystal may include two or more types of non-lamellar liquid crystal phases.

[0060] The cubic liquid crystal may be a cubic liquid crystal belonging to the crystallographic space group Ia3d (hereinafter referred to as Ia3d cubic liquid crystal), a cubic liquid crystal belonging to the crystallographic space group Pn3m (hereinafter referred to as Pn3m cubic liquid crystal), or a cubic liquid crystal belonging to the crystallographic space group Im3m (hereinafter referred to as Im3m cubic liquid crystal).

[0061] The liquid crystal structure can be analyzed by a conventional method, for example, by small angle X-ray scattering (SAXS) measurement using the following method.

[0062] When the measurement sample is an emulsion, for example, the sample may be placed in an X-ray capillary tube made of soda glass or quartz, the capillary may be sealed with an oxygen burner, and then subjected to SAXS measurement. SAXS measurement can be performed using a commercially available instrument, for example, a NANO Viewer nanoscale X-ray structure evaluation instrument (manufactured by Rigaku).

[0063] By checking whether the SAXS measurement results show the following scattering peak ratio (peak interval) specific to each liquid crystal structure, it is possible to confirm whether the non-lamellar liquid crystal forming composition of the present invention has formed or will form in the presence of water. Ratios for Pn3m cubic liquid crystals: √2:√3:√4:√6:√8:√9:√10: ,,,, Ratios for Ia3d cubic liquid crystals: √3:√4:√7:√8:√10:√11: ,,,, Ratios for Im3m cubic liquid crystals: √2:√4:√6:√8:√10:√12:√14: ,,,, Ratios for Fd3m cubic liquid crystals: √3:√8:√11:√12:√16:√19:√24:√27: ,,,, Ratios specific to inverted hexagonal liquid crystals: 1:√3:2: ,,,, Furthermore, the space group and lattice constants can be easily determined by calculating peak values ​​from SAXS intensity distribution data and then calculating the ratio of their reciprocals according to methods well known to those skilled in the art.

[0064] On the other hand, in the SAXS measurement of the sponge phase (L3 phase), a broad scattering peak is observed.

[0065] The scattering vector value q1 [nm －１ By analyzing the scattering vector values ​​of peaks in the measurement sample, including

[0049] , it is possible to determine not only the type of liquid crystal phase but also its interplanar spacing and lattice constant. Based on the results of such analysis, the size of the liquid crystal phase and unit lattice can be changed by changing the composition of the non-lamellar liquid crystal-forming composition, thereby controlling the sustained release (sustained release rate) of a drug from a composition or formulation (e.g., precursor formulation, emulsion) containing the non-lamellar liquid crystal-forming composition and a drug. For example, by changing the size of the liquid crystal phase and unit lattice depending on the fatty acid ester, phospholipid, and other components, and their ratios (weight ratios), a corresponding sustained release rate can be obtained. Typically, the sustained release rate increases in the following order (from lowest to highest): reverse micellar cubic phase (Fd3m), reverse hexagonal liquid crystal (HII), and cubic liquid crystal.

[0066] The non-lamellar liquid crystal forming composition according to the present invention can be used as a medical substrate for application to the body. The non-lamellar liquid crystal forming composition according to the present invention has an anti-adhesion effect on biological tissues, and can therefore be used to prevent adhesion of biological tissues. Therefore, the non-lamellar liquid crystal forming composition according to the present invention may be used for preventing adhesion of biological tissues. The non-lamellar liquid crystal forming composition according to the present invention can be applied (administered) in vivo as an agent for preventing adhesion of biological tissues.

[0067] The non-lamellar liquid crystal forming composition of the present invention can prevent adhesion of biological tissues by applying it to biological tissues that are prone to adhesion. In the present invention, the "adhesion prevention effect" refers to the effect of preventing adhesion of tissues that are prone to adhesion to other tissues or organs, making them difficult to separate, and completely or at a low level of adhesion. Therefore, the non-lamellar liquid crystal forming composition of the present invention can be used to prevent adhesion of biological tissues.

[0068] The adhesion prevention effect of the non-lamellar liquid crystal forming composition of the present invention is achieved by the formation of a coating on the surface of the tissue to which the non-lamellar liquid crystal forming composition is applied, due to the formation of non-lamellar liquid crystals by the amphiphilic compound (fatty acid ester). The formed coating prevents contact between the tissue and other tissues or organs, thereby reducing adhesion.

[0069] The adhesion prevention effect of the nonlamellar liquid crystal-forming composition according to the present invention can be confirmed, for example, by applying the nonlamellar liquid crystal-forming composition to the tissue incision in an animal model after abdominal opening, closing the abdomen, and then observing the progress. Specifically, a rat is subjected to a midline abdominal incision (e.g., approximately 30 mm), and an approximately 20 mm incision is made in the left and right upper abdominal parietal peritoneum. After complete hemostasis, the peritoneal incision is closed with continuous sutures (e.g., using 5-0 silk suture). A nonlamellar liquid crystal-forming composition sample is then applied to either the left or right peritoneal incision so as to cover the sutured incision. When the nonlamellar liquid crystal-forming composition is a liquid crystal precursor composition, an aqueous medium (e.g., water for injection) may be further added to the sample application site by spraying, etc., to induce liquid crystal formation. No medium is applied to the other peritoneal incision. The abdominal wall is then closed in two layers. After a certain period of time (e.g., 7 days) following this surgery, the abdomen is opened and the presence of adhesions at the sutured incision is evaluated.

[0070] The application of the non-lamellar liquid crystal forming composition may be carried out in a manner appropriate for the dosage form. The amount of the non-lamellar liquid crystal forming composition applied in the above evaluation is typically preferably an amount equivalent to 5 to 50 mg of the amphiphilic compound.

[0071] Adhesion evaluation can be performed, for example, by assigning an evaluation score regarding adhesion strength as follows.

[0072] Grade 0: No adhesions Grade 1: Adhesion that can be separated with light traction (no tissue damage) Grade 2: Adhesion that can be separated with strong traction (no tissue damage) Grade 3: Adhesion that involves tissue damage when separated with strong traction If the evaluation score is lower in the incision site to which the non-lamellar liquid crystal forming composition sample was applied compared to the incision site to which the non-lamellar liquid crystal forming composition sample was not applied on the same animal, it can be determined that an adhesion prevention effect is observed.

[0073] Furthermore, for example, the adhesion range rate of each incision site is calculated as the ratio (%) of the adhesion length to the approximately 20 mm incision suture site, and based on this, the ratio of the adhesion range of the incision site to which the non-lamellar liquid crystal forming composition sample was applied to the incision site to which the sample was not applied (adhesion range rate on the sample-applied side / adhesion range rate on the non-applied side × 100) can be judged as A if it is 40% or less, B+ if it is 41 to 60%, B- if it is 61 to 80%, and C if it is 81% or more. In this case, a rating of A or B+ is preferable in terms of adhesion prevention effect.

[0074] In the present invention, "adhesion prevention" can be determined by the reduction in the frequency and / or severity of adhesions at the application site by application (treatment) of the non-lamellar liquid crystal forming composition compared to an untreated control.

[0075] The present invention also provides a method for preventing adhesions in biological tissues, comprising applying the non-lamellar liquid crystal-forming composition of the present invention to biological tissues. More specifically, the present invention also provides a method for preventing tissue adhesions in an affected area, comprising applying an effective amount of the non-lamellar liquid crystal-forming composition of the present invention to an affected area of ​​a patient, specifically a site at risk of adhesions, specifically a site where tissue repair is expected to occur (e.g., an inflamed or damaged site within the body). Specific examples of such sites at risk of adhesions include sites of exogenous or endogenous inflammation within the body, wound sites such as surgical incisions, and sites where the tissue surface has been damaged by artificial manipulation such as touching during surgery. In the present invention, the term "damaged site" refers to a portion of a tissue or organ damaged by surgery, trauma, disease, or the like. Examples of tissues or organs to which the adhesion inhibitor can be applied include, but are not limited to, the peritoneum, small intestine, large intestine, rectum, stomach, duodenum, cecum, liver, uterus, fallopian tubes, lymphatic vessels, heart, pericardium, lungs, brain, ovaries, and tendons. In a typical example, the non-lamellar liquid crystal forming composition of the present invention is applied to an incision site, the area around an incision site, or the entire organ having an incision site during surgery. The non-lamellar liquid crystal forming composition of the present invention may also be applied to an internal site that comes into contact with a wound site, an inflamed site, or the like.

[0076] Application to affected areas such as injury sites (e.g., wound sites) and inflammation sites can be performed using a method appropriate for the dosage form. For example, the adhesion inhibitor can be sprayed onto affected areas such as injury sites (e.g., wound sites) and inflammation sites using a gas-propelled aerosol container. Furthermore, in the case of a pump spray, the non-lamellar liquid crystal-forming composition can be sprayed onto affected areas such as injury sites (e.g., wound sites) and inflammation sites using a non-gas-propelled spray container, such as a general-purpose manual type. In the case of endoscopic surgery or laparoscopic surgery, the non-lamellar liquid crystal-forming composition can also be sprayed onto affected areas such as injury sites (e.g., wound sites) using a spray nozzle or the like used in endoscopic surgery or laparoscopic surgery. In the present invention, "spraying" refers to applying pressure to eject (atomize and / or jet) the target substance in the form of droplets, mist, fine particles, foam, or the like. When the non-lamellar liquid crystal-forming composition is a liniment, an appropriate amount can be applied to affected areas such as injury sites (e.g., wound sites) and inflammation sites. When the non-lamellar liquid crystal forming composition is an injection, the non-lamellar liquid crystal forming composition may be injected into an affected area such as an injured site (for example, a wound site) or an inflamed site.

[0077] The non-lamellar liquid crystal forming composition according to the present invention is preferably applied to an affected area such as an injured area (e.g., a wound area) or an inflamed area in an amount sufficient to sufficiently cover the affected area such as an injured area (e.g., a wound area) or an inflamed area. In a preferred embodiment, the specific application amount of the non-lamellar liquid crystal forming composition according to the present invention is 10 mg to 100 g, more preferably 50 mg to 50 g, and particularly preferably 0.1 g to 10 g, for humans.

[0078] When the non-lamellar liquid crystal-forming composition of the present invention contains a sufficient amount of aqueous medium (e.g., when it is a liquid crystal emulsion), the non-lamellar liquid crystal-forming composition can form non-lamellar liquid crystals on the surface of tissue to which it is applied. When the non-lamellar liquid crystal-forming composition of the present invention does not contain a sufficient amount of aqueous medium (e.g., when it is a liquid crystal precursor composition), non-lamellar liquid crystals are formed by moisture in the body. However, to promote film formation, it is preferable to apply an aqueous medium in addition to the non-lamellar liquid crystal-forming composition to an affected area such as an injury site (e.g., a wound site) or an inflammation site. The aqueous medium may be, for example, water such as sterilized water, purified water, distilled water, ion-exchanged water, ultrapure water, or water for injection, or a physiologically acceptable aqueous solution. Examples of physiologically acceptable aqueous solutions include physiological saline; electrolyte solutions such as 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; buffer solutions such as phosphate buffer and Tris-HCl buffer; aqueous solutions containing sugar molecules such as glucose, sucrose, maltose, and hyaluronic acid; and aqueous solutions containing water-soluble polymers such as polyethylene glycol and polyvinyl alcohol. A preferred example of a physiologically acceptable aqueous solution is a hyaluronic acid aqueous solution containing hyaluronic acid or a salt thereof (such as sodium hyaluronate).

[0079] It is preferable, but not limited to, to apply an aqueous medium onto the non-lamellar liquid crystal-forming composition after applying the non-lamellar liquid crystal-forming composition, which is a liquid crystal precursor composition. The aqueous medium can be applied by the same application method as for the non-lamellar liquid crystal-forming composition, for example, by spraying, coating, or injection. After applying the aqueous medium to the tissue or organ in this manner, it is preferable to leave it as is for a predetermined time (for example, but not limited to, 1 to 30 minutes, preferably 5 to 10 minutes) to promote film formation.

[0080] The subject (patient) to whom the adhesion prevention method using the non-lamellar liquid crystal forming composition of the present invention is applied is typically a mammal such as a human, livestock, pet animal, laboratory animal, etc. Particularly preferred is a subject whose tissue (organ) has been or is expected to be damaged by surgery, trauma, disease, etc. Surgical operations include not only open surgery, but also endoscopic surgery and laparoscopic surgery.

[0081] The non-lamellar liquid crystal forming composition of the present invention has significantly reduced or no toxicity, and therefore the adhesion prevention method according to the present invention is highly safe for patients undergoing treatment.

[0082] The non-lamellar liquid crystal forming composition of the present invention may contain a drug in addition to the above-mentioned components, such as the fatty acid ester and phospholipid. In the present invention, the drug refers to any substance (active ingredient) to be administered to a living body so that it can be retained in the non-lamellar liquid crystal structure and sustainedly (controlled) released by inclusion in the non-lamellar liquid crystal forming composition. However, the drug is not the fatty acid ester itself. The drug may be an organic compound or an inorganic compound. The drug may be a water-soluble drug or a lipid-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 to these. The drug may be, for example, a gonadotropin-releasing hormone (GnRH) agonist. The gonadotropin-releasing hormone (GnRH) agonist may be, for example, but is not limited to, leuprolide or a salt thereof. The salt of leuprolide may be any pharmaceutically acceptable salt, including, but not limited to, carboxylic acid salts such as acetate (i.e., leuprolide acetate). Leuprolide acetate is also known by other names such as leuprorelin acetate. Such non-lamellar liquid crystal forming compositions can be used for sustained release of drugs.

[0083] The present invention also provides a pharmaceutical formulation comprising the non-lamellar liquid crystal-forming composition of the present invention. The pharmaceutical formulation of the present invention comprises a non-lamellar liquid crystal-forming composition comprising the fatty acid ester and a phospholipid, the biocompatibility of which is improved by the phospholipid. In one embodiment, the pharmaceutical formulation of the present invention comprises a non-lamellar liquid crystal-forming composition comprising the fatty acid ester, a phospholipid, and a drug. The "pharmaceutical formulation" of the present invention may be a pharmaceutical composition. The pharmaceutical formulation or pharmaceutical composition of the present invention may further comprise other substances such as pharmaceutically acceptable additives (e.g., carriers, excipients, lubricants, disintegrants, humectants, buffers, flavoring agents, preservatives, colorants, fragrances, propellants, etc.), as long as the pharmaceutical formulation or pharmaceutical composition can maintain its ability to form non-lamellar liquid crystals.

[0084] The pharmaceutical preparation according to the present invention may be formulated into any dosage form, such as a spray, an aerosol, an injection, or a depot.

[0085] The pharmaceutical preparation of the present invention may be used to prevent adhesion of biological tissues. The pharmaceutical preparation of the present invention may also be a sustained-release preparation such as a depot preparation, which further contains the above-mentioned drug.

[0086] The present invention also provides a method for sustained delivery of a drug into a living organism (body) or living cells or tissues, comprising applying a pharmaceutical formulation containing the drug of the present invention or a non-lamellar liquid crystal-forming composition containing the drug of the present invention to a living organism such as a subject (patient), for example, the body (particularly, a living tissue within the body) or body surface, or to living cells or tissues. When using a non-lamellar liquid crystal-forming composition that is a liquid crystal precursor composition or a pharmaceutical formulation containing the same, an aqueous medium may be applied over the non-lamellar liquid crystal-forming composition or pharmaceutical formulation after application, but this is not limited thereto; liquid crystal formation due to water in the body may also be utilized. Here, application to a living organism (such as the body or body surface) or living cells or tissues is preferably carried out parenterally (e.g., intravenously, intra-arterially, intraperitoneally, intramuscularly, subcutaneously, or intradermally). The method of the present invention enables sustained delivery of a drug into the body or living cells or tissues with high safety.

[0087] The present invention will be described in more detail below using examples, although the technical scope of the present invention is not limited to these examples.

[0088] Example 1 Synthesis of amphiphilic compounds (fatty acid esters) (1) Synthesis of propylene glycol monooleate

[0089]

[0090] Under reduced pressure of 8-9 kPa, 46.6 g of ethyl oleate (150 mmol, Olive Ethyl Oleate NIKKOL EOO, Nikko Chemicals Co., Ltd.; typical composition of the main fatty acids in olive oil: oleic acid 75%, palmitic acid 12%, linoleic acid 7%, stearic acid 3%, palmitoleic acid 1%) was added dropwise to a solution of 45.6 g of propylene glycol (600 mmol) and 290 mg of potassium carbonate (2.10 mmol) in dry N,N-dimethylformamide (105 mL) at 85°C over 1 hour, followed by stirring at the same temperature for 2 hours. Further, 83 mg of potassium carbonate (0.60 mmol) was added, followed by stirring at the same temperature for 1 hour. The reaction solution was cooled to 65°C and neutralized with 283 μL of formic acid (7.5 mmol). The pressure was then reduced to 1-2 kPa, and the N,N-dimethylformamide was distilled off. The resulting solution was diluted with an ethyl acetate / hexane mixed solvent (1:1, 140 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 (mobile phase: hexane / ethyl acetate = 100:0 to 40:60) to obtain 41.5 g (yield 81%) of the title compound as a pale yellow, transparent liquid with low viscosity. The obtained compound was １ The results of H-NMR measurement are as follows:

[0091] １ H-NMR spectrum (300 MHz, CDCl ３ ​, TMS) δ: 0.88 (m, 3H), 1.2-1.4 (m, 23H), 1.64 (m, 2H), 1.95-2.15 (m, 4H), 2.26-2. 38 (m, 2H), 3.55-3.70 (m, 0.7H), 3.9-4.15 (m, 2.0H), 4.99 (m, 0.3H), 5.35 (m, 2H)

[0092] Propylene glycol monooleate is also called propylene glycol monooleate (C18:1).

[0093] (2) Synthesis of propylene glycol monomyristate

[0094]

[0095] A solution of 2.70 g (10.0 mmol) of isopropyl myristate, 2.43 g (31.9 mmol) of propylene glycol, and 25 mg (0.18 mmol) of potassium carbonate in dry N,N-dimethylformamide (7.0 mL) was stirred at 85°C for 8 hours under reduced pressure of 8 to 9 kPa. The reaction solution was cooled to room temperature and then neutralized with 19 μL (0.50 mmol) of formic acid. The resulting solution was diluted with an ethyl acetate / hexane mixed solvent (1:1, 35 mL), washed with water, saturated sodium bicarbonate water, and saturated saline (twice), and then dried over magnesium sulfate. After filtration and concentration, 2.80 g (98% yield) of the title compound was obtained as a colorless, transparent liquid with low viscosity. The resulting compound was analyzed as follows: １ The results of H-NMR measurement are as follows:

[0096] １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.88 (t, J=6.7Hz, 3H), 1.2-1.4 (m, 23H), 1.63 (m, 2H), 2.26-2.38 (m, 2H), 3.56-3.72 (m, 0.7H), 3.9-4.2 (m, 1.9H), 5.0 (m, 0.4H)

[0097] Propylene glycol monomyristate is also called propylene glycol monomyristate (C14:0).

[0098] (3) Synthesis of propylene glycol monopalmitate ​

[0099]

[0100] The same procedure as in Example 1(2) was carried out except that 2.70 g (10.0 mmol) of isopropyl myristate in Example 1(2) was replaced with 2.99 g (10.0 mmol) of isopropyl palmitate to obtain 3.12 g (yield 99%) of the title compound as a waxy solid. １ The results of H-NMR measurement are as follows:

[0101] １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.88 (t, J=6.6Hz, 3H), 1.2-1.4 (m, 27H), 1.62 (m, 2H), 2.26-2.38 (m, 2H), 3.56-3.72 (m, 0.7H), 3.9-4.2 (m, 1.9H), 5.0 (m, 0.4H)

[0102] Propylene glycol monopalmitate is also called propylene glycol monopalmitate (C16:0).

[0103] (4) Synthesis of propylene glycol monolinoleate

[0104]

[0105] A solution of 3.09 g (10.0 mmol) of ethyl linoleate, 2.43 g (31.9 mmol) of propylene glycol, and 25 mg (0.18 mmol) of potassium carbonate in dry N,N-dimethylformamide (7.0 mL) was stirred at 85°C for 2 hours under reduced pressure of 8 to 9 kPa. The reaction solution was cooled to room temperature and neutralized with 19 μL (0.50 mmol) of formic acid. The resulting solution was diluted with an ethyl acetate / hexane mixed solvent (1:1, 35 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 (mobile phase: hexane / ethyl acetate = 100:0 to 40:60) to obtain 2.45 g (72% yield) of the title compound as a colorless, transparent liquid with low viscosity. The resulting compound was characterized as follows: １ ​The results of H-NMR measurement are as follows:

[0106] １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.89 (m, 3H), 1.2-1.4 (m, 17H), 1.63 (m, 2H), 1.95-2.10 (m, 4H), 2.30-2.38 (m, 2H) , 2.77 (m, 2H), 3.55-3.70 (m, 0.7H), 3.89-4.14 (m, 2.0H), 5.0 (m, 0.3H), 5.27-5.44 (m, 4H)

[0107] Propylene glycol monolinoleate is also called propylene glycol monolinoleate (C18:2).

[0108] (5) Synthesis of propylene glycol monobehenate

[0109]

[0110] Under a nitrogen atmosphere, 1.3 mL (15 mmol) of oxalyl chloride was added dropwise to a solution of 3.41 g (10.0 mmol) of behenic acid in dry methylene chloride (30 mL) at room temperature. The mixture was stirred at the same temperature for 2 hours and then concentrated under reduced pressure. A solution of the resulting residue in dry methylene chloride (20 mL) was added to a solution of 1.14 g (15.0 mmol) of propylene glycol and 1.61 mL (20.0 mmol) of pyridine in dry N,N-dimethylformamide (20 mL) at 0°C under a nitrogen atmosphere, and the mixture was immediately stirred overnight at room temperature. The resulting reaction solution was diluted with an ethyl acetate / hexane mixed solvent (1:1, 60 mL), washed with water, 1 M hydrochloric acid, saturated aqueous sodium bicarbonate, and saturated saline, and then dried over magnesium sulfate. After filtration and concentration, 3.82 g (yield 96%) of the title compound was obtained as a white powder. Regarding the resulting compound, １ The results of H-NMR measurement are as follows:

[0111] １ H-NMR spectrum (300 MHz, CDCl ３ ​​, TMS) δ: 0.88 (t, J=6.6Hz, 3H), 1.2-1.4 (m, 39H), 1.63 (m, 2H), 2.26-2.38 (m, 2H), 3.58-3.72 (m, 0.4H), 3.89-4.19 (m, 2.4H), 5.0 (m, 0.2H)

[0112] Propylene glycol monobehenate is also called propylene glycol monobehenate (C22:0).

[0113] (6) Synthesis of ethylene glycol monooleate

[0114]

[0115] The same procedure as in Example 1(4) was carried out except that 3.09 g (10.0 mmol) of ethyl linoleate and 2.43 g (31.9 mmol) of propylene glycol in Example 1(4) were replaced with 3.11 g (10.0 mmol) of ethyl oleate and 1.98 g (31.9 mmol) of ethylene glycol, to obtain 2.88 g (yield 88%) of the title compound as a colorless, transparent liquid. １ The results of H-NMR measurement are as follows:

[0116] １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.88 (m, 3H), 1.2-1.4 (m, 20H), 1.63 (m, 2H), 1.95-2.1 (m, 4H), 2.35 (t, J = 7.6Hz, 2H), 3.83 (m, 2H), 4.21 (m, 2H), 5.34 (m, 2H)

[0117] Ethylene glycol monooleate is also called ethylene glycol monooleate (C18:1).

[0118] (7) Synthesis of 1,3-butylene glycol monooleate

[0119]

[0120] ​The same procedure as in Example 1(4) was carried out, except that 3.09 g (10.0 mmol) of ethyl linoleate and 2.43 g (31.9 mmol) of propylene glycol in Example 1(4) were replaced with 3.11 g (10.0 mmol) of ethyl oleate and 2.87 g (31.9 mmol) of 1,3-butylene glycol, to obtain 2.90 g (yield 82%) of the title compound as a colorless, transparent liquid with low viscosity. １ The results of H-NMR measurement are as follows:

[0121] １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.88 (m, 3H), 1.2-1.4 (m, 23H), 1.62 (m, 2H), 1.6-1.9 (m, 2H), 1.95-2.1 (m, 4H), 2.26-2.34 (m, 2H), 3.5-3. 7 (m, 0.5H), 3.86 (m, 0.75H), 4.12 (m, 0.75H), 4.35 (ddd, J=5.3, 8.3, 13.6Hz, 0.75H), 5.12 (m, 0.25H), 5.34 (m, 2H)

[0122] 1,3-butylene glycol monooleate is also called butylene glycol monooleate (C18:1).

[0123] (8) Synthesis of 3-methyl-1,3-butanediol monooleate

[0124]

[0125] The same procedure as in Example 1(4) was carried out, except that 3.09 g (10.0 mmol) of ethyl linoleate and 2.43 g (31.9 mmol) of propylene glycol in Example 1(4) were replaced with 3.11 g (10.0 mmol) of ethyl oleate and 3.32 g (31.9 mmol) of 3-methyl-1,3-butanediol, to obtain 2.31 g (yield 63%) of the title compound as a colorless, transparent liquid with low viscosity. １ The results of H-NMR measurement are as follows:

[0126] １ H-NMR spectrum (300 MHz, CDCl​​３ , TMS) δ: 0.88 (m, 3H), 1.2-1.4 (m, 26H), 1.61 (m, 2H), 1.84 (t, J=6.9Hz, 2H), 1.95-2.1 (m, 4H), 2.30 (t, J = 7.5Hz, 2H), 4.25 (t, J = 6.9Hz, 2H), 5.34 (m, 2H)

[0127] 3-Methyl-1,3-butanediol monooleate is also called isoprene glycol monooleate (C18:1).

[0128] (9) Synthesis of diethylene glycol monooleate

[0129]

[0130] The same procedure as in Example 1(4) above (0.08 equivalents of potassium carbonate was added) was carried out, except that 3.09 g (10.0 mmol) of ethyl linoleate and 2.43 g (31.9 mmol) of propylene glycol in Example 1(4) were replaced with 3.11 g (10.0 mmol) of ethyl oleate and 3.39 g (31.9 mmol) of diethylene glycol, to obtain 2.57 g (yield 69%) of the title compound as a pale yellow, transparent liquid with low viscosity. １ The results of H-NMR measurement are as follows:

[0131] １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.88 (m, 3H), 1.2-1.4 (m, 20H), 1.63 (m, 2H), 1.95-2.1 (m, 4H), 2.28- 2.38 (m, 2H), 3.61 (m, 2H), 3.71 (m, 2H), 3.74 (m, 2H), 4.25 (m, 2H), 5.34 (m, 2H)

[0132] Diethylene glycol monooleate is also called diethylene glycol monooleate (C18:1).

[0133] ​Example 2 Preparation of Precursor Formulation, Gel Formation Test, and Analysis of Liquid Crystal Structure According to the blending ratios shown in Tables 1 and 2 below, the amphiphilic compound (fatty acid ester) in which the hydrophilic group has one hydroxyl group was the amphiphilic compound synthesized in Example 1 or monocaprylic acid (C8:0) propylene glycol (propylene glycol monocaprylate) (NIKKOL SEFSOL-218, Nikko Chemicals Co., Ltd.), and as phospholipids, soybean phosphatidylcholine (soybean lecithin, LIPOID S100, Lipoid Co., Ltd.; abbreviation SPC is used below and in the tables), egg yolk phosphatidylcholine (purified egg yolk lecithin, PL-100M, Kewpie Corporation; abbreviation EPC is used below and in the tables), or dioleylphosphatidylethanolamine (COATSOME ME-8181, NOF Corporation. The abbreviation DOPE is used below and in the tables), sesame oil (triglyceride, Japanese Pharmacopoeia Sesame Oil, Kaneda Co., Ltd.) as an oil component, and EtOH (ethanol) were mixed. The resulting mixtures were dissolved in a water bath at 40°C or less to prepare precursor formulations No. 1 to 23 shown in Tables 1 and 2.

[0134] A gel formation test was conducted on precursor formulations No. 1 to 23. A portion of each precursor formulation (approximately 100 to 300 mg) was added to an excess amount of water for injection (approximately 0.5 to 2 mL) in a vial, and the mixture was mixed at room temperature (25°C) using a spoon and / or a vortex mixer. As a result, for all precursor formulations No. 1 to 23, a gel composition that was colorless, transparent, or cloudy in appearance was obtained, which separated into the excess water for injection (aqueous medium).

[0135] The gel compositions obtained from the precursor preparations Nos. 1 to 23 were embedded directly into a pinhole slit, and the non-lamellar liquid crystal structure was analyzed by small-angle X-ray scattering diffraction measurement using a small-angle X-ray scattering (SAXS) device (Nano-Viewer, manufactured by Rigaku Corporation).

[0136] The obtained liquid crystal phase and the scattering vector value q1 [nm －１] are shown in Tables 1 and 2. HII means reverse hexagonal liquid crystal, Pn3m means reverse cubic liquid crystal belonging to the crystallographic space group Pn3m, Im3m means reverse cubic liquid crystal belonging to the crystallographic space group Im3m, and Fd3m means reverse cubic liquid crystal belonging to the crystallographic space group Fd3m. Some gel compositions had two different liquid crystal phases.

[0137] Among the amphiphilic compounds used, in which the hydrophilic group has one hydroxyl group, diethylene glycol monooleate alone formed a gel composition when mixed with water (liquid crystal phase: H1I, q1: 1.35 nm). －１ The amphiphilic compounds other than diethylene glycol monooleate did not form gel compositions by themselves when mixed with water.

[0138] As shown in Tables 1 and 2, it was suggested that the precursor formulation was capable of forming non-lamellar liquid crystals over a wide range of weight ratios of amphiphilic compound with one hydroxyl group in the hydrophilic group to phospholipid. Furthermore, the precursor formulation was also capable of forming non-lamellar liquid crystals over a wide range of weight ratios of amphiphilic compound + phospholipid (total amount of amphiphilic compound and phospholipid):oil. Various combinations of amphiphilic compound, phospholipid, and oil resulted in the formation of a variety of liquid crystal structures, such as reversed cubic liquid crystals and reversed hexagonal liquid crystals belonging to the crystallographic space groups Pn3m, Im3m, or Fd3m, and the q1 value due to the lattice constant varied over a very wide range of values.

[0139]

[0140]

[0141] Example 3: Preparation of Emulsion (Dispersant) and Analysis of Liquid Crystal Structure and Particle Size Distribution. Monooleic acid (C18:1) propylene glycol (synthesized in Example 1) as an amphiphilic compound with one hydroxyl group in the hydrophilic group, SPC as a phospholipid, sesame oil as an oil, and EtOH were mixed in the proportions shown in Table 3 below to obtain an oily solution. Meanwhile, Pluronic F127 (Unilube® 70DP-950B, NOF Corporation) as a surfactant and water for injection (Otsuka Distilled Water) were mixed to obtain an aqueous Pluronic solution. The oily solution and the aqueous Pluronic solution thus prepared were each completely dissolved in a water bath at 40°C or less, then mixed together at room temperature and stirred with a spoon or stirrer tip to form a suspension. This suspension was then dispersed using a high-pressure homogenizer (Starbust Minimo, manufactured by Sugino Machine) to prepare white emulsions containing fine particles (Formulations No. 24 and 25). These emulsions were prepared in 8 g quantities each.

[0142] The emulsions of Formulation Nos. 24 and 25 thus obtained were subjected to structural analysis by small-angle X-ray scattering (SAXS) using a NANO Viewer nanoscale X-ray structural evaluation system (manufactured by Rigaku). Each emulsion was introduced into a capillary at atmospheric pressure and measured in a reduced pressure apparatus (the sample itself was at atmospheric pressure). At least three scattering peaks were observed in the scattering intensity distribution obtained from the emulsion of Formulation No. 24. The peak ratio was 1:√3:2 (q1:1.22 nm), which is characteristic of inverted hexagonal liquid crystals. －１ ), indicating that this emulsion is a liquid crystal emulsion (hexasome) in which reverse hexagonal liquid crystal particles are dispersed in an aqueous phase. A broad scattering peak was observed in the scattering intensity distribution obtained from the emulsion of Formulation No. 25, and these emulsions were considered to be liquid crystal emulsions in which sponge phase (L3 phase) particles are dispersed in an aqueous phase.

[0143] Furthermore, the particle size distribution of the emulsions of Formulations No. 24 and 25 was measured by dynamic light scattering using a Zetasizer Nano-ZS (Malvern Instruments). Measurement samples were prepared by diluting each emulsion 200 times with distilled water immediately after preparation (within 4 days at room temperature). Table 3 shows the average particle size (nm) (Z-Average) and PDI (polydispersity index) obtained from each measurement sample. After 3 months at room temperature, each emulsion was visually stable.

[0144] Liquid crystal emulsions having stable non-lamellar liquid crystals over time could be prepared in a wide range of amphiphilic compound:phospholipid ratios and amphiphilic compound + phospholipid (total amount of amphiphilic compound and phospholipid):oil ratios shown in Table 3.

[0145]

[0146] Example 4 Preparation, property confirmation, gel formation test, and viscoelasticity test of precursor formulations using amphipathic compounds with different numbers of hydroxyl groups in the hydrophilic group The properties of precursor formulations containing phospholipids were compared for amphipathic compounds with hydrophilic groups having one to three hydroxyl groups.

[0147] The amphiphilic compound used was propylene glycol monooleate (C18:1) (synthesized in Example 1) as the amphiphilic compound having one hydroxyl group in the hydrophilic group, glyceryl monooleate (Rikemal XO-100, NOF Corporation) as the amphiphilic compound having two hydroxyl groups in the hydrophilic group, and purified sorbitan monooleate (sorbitan monooleate NIKKOL SO-10V (Nikko Chemicals Co., Ltd.) obtained by removing low-polarity components such as oleic acid and sorbitan dioleate by silica gel column purification) as the amphiphilic compound having three hydroxyl groups in the hydrophilic group. Note that propylene glycol monooleate (C18:1) is a light yellow, transparent liquid with low viscosity, whereas glyceryl monooleate is a waxy solid and purified sorbitan monooleate is a highly viscous fluid.

[0148] According to the compounding ratios shown in Table 4 below, 2.2 g of each of precursor preparations Nos. 26 to 31 was prepared by mixing and dissolving in the same manner as in Example 2.

[0149] The precursor formulations No. 26 and 27, which used propylene glycol monooleate (C18:1), could be prepared in a shorter time than the precursor formulations No. 28 and 30, which had a different amphipathic compound but an amphipathic compound ratio of 60:40, or the precursor formulations No. 29 and 31, which had a different amphipathic compound but an amphipathic compound ratio of 40:60. The preparation time for the precursor formulations No. 26 and 27 was within one hour, while the preparation time for the precursor formulations No. 28 to 31 was approximately two hours. The precursor formulations No. 28 to 31, which used glyceryl monooleate or purified sorbitan monooleate, could not be uniformly dissolved without the use of an organic solvent (e.g., EtOH), and therefore could not be prepared. The precursor formulations No. 29 to 31, which used propylene glycol monooleate (C18:1), could not be uniformly dissolved without the use of an organic solvent (e.g., EtOH). The precursor preparation of No. 26 could be uniformly dissolved without using an organic solvent, making it possible to prepare a precursor preparation (see Preparation No. 3 in Example 2).

[0150] Furthermore, in the precursor formulation No. 28, which used glyceryl monooleate and had an amphiphilic compound:phospholipid ratio of 60:40, glyceryl monooleate began to precipitate at around 20°C and solidified when stored in the refrigerator.

[0151] A gel formation test was performed on precursor formulations No. 26 to 31 in the same manner as in Example 2. As a result, precursor formulations No. 26 to 28, 30, and 31 yielded gel compositions that separated in excess water for injection (aqueous medium) and were colorless, transparent, or cloudy in appearance (the small amount of EtOH had little effect on the non-lamellar liquid crystal structure, and the non-lamellar liquid crystal structures obtained from precursor formulations No. 2, 3, and 26 and those from precursor formulations No. 5 and 27 were almost identical). On the other hand, precursor formulation No. 29, which used glyceryl monooleate and had an amphiphilic compound:phospholipid ratio of 40:60, did not yield a gel composition, and the entire solution in the vial became emulsion-like.

[0152] For precursor formulations No. 26 to 31, shear viscosity measurements were performed using a viscosity / viscoelasticity measuring device (MCR302, Anton Paar; cone plate φ50, cone angle 1°, temperature 23 to 25°C). Table 4 shows the viscosities [Pa s] at shear rates of 1 (1 / s) and 100 (1 / s).

[0153] When comparing precursor formulations No. 26, 28, and 30, which had an amphiphilic compound:phospholipid ratio of 60:40, and precursor formulations No. 27, 29, and 31, which had an amphiphilic compound:phospholipid ratio of 40:60, the precursor formulations using propylene glycol monooleate (C18:1) (precursor formulations No. 26 and 27, respectively) had the lowest viscosity at both shear rates of 1 (1 / s) and 100 (1 / s). Even precursor formulation No. 27, which had a more viscous amphiphilic compound:phospholipid ratio of 40:60, had a lower viscosity than precursor formulations No. 28 to 31, which were made with other lipids.

[0154] These results demonstrate that precursor formulations containing an amphiphilic compound with one hydroxyl group in the hydrophilic group and a phospholipid (wherein the fatty acid of the amphiphilic compound is oleic acid) are easier to prepare and more stable, more likely to form non-lamellar liquid crystal structures, and have significantly lower viscosity than precursor formulations containing an amphiphilic compound with two or three hydroxyl groups in the hydrophilic group and a phospholipid (wherein the fatty acid of the amphiphilic compound is oleic acid). A lower viscosity of the composition is advantageous, for example, because it allows for thinner needles when used as an injection.

[0155]

[0156] Example 5 Preparation of Precursor Formulations Formulated with Leuprolide Acetate and In Vitro Release Testing Propylene glycol monooleate (C18:1) as an amphiphilic compound having one hydroxyl group as the hydrophilic group, SPC as a phospholipid, sesame oil as an oil, and EtOH were mixed according to the blending ratios shown in Table 5 below, and then dissolved in a water bath at 40°C or lower. Next, a dimethyl sulfoxide (DMSO) solution of leuprolide acetate (L0249, Tokyo Chemical Industry Co., Ltd.; hereinafter and in the tables, abbreviated as LA) was added and dissolved uniformly, thereby obtaining precursor formulations Nos. 32 to 35 each containing leuprolide acetate.

[0157] An in vitro release test was conducted on precursor formulations No. 32, 33, and 35 (100 mg) containing leuprolide acetate. A dialysis tube with a floating rack attached to the top was placed in a 25 mL vial containing 20 mL of PBS solution at pH 7.4 containing 0.02% P80 (polyoxyethylene sorbitan monooleate (20E.O.)) so that the precursor formulation was fully immersed in the PBS solution. The tube was then left to stand at room temperature (25°C). Subsequently, 500 μL of the PBS solution in the vial was sampled at 0, 1, 3, 6, 12, 24, 48, 72, 120, and 168 hours after the start of the test over a 7-day period. It was visually observed that precursor formulations No. 32, 33, and 35, when added to the PBS solution, all formed gel compositions.

[0158] The quantification of leuprolide acetate in the sample collected from the PBS solution was carried out by LC / MS / MS analysis using a previously prepared calibration curve under the following analytical conditions:

[0159] Analytical column: Shodex ODP2HP-2B 2.0 mm I.D. x 50 mm (Showa Denko K.K.) Mobile phase: water (containing 0.1% formic acid):acetonitrile = 70:30 Flow rate: 0.1 mL / min, column temperature: 40°C, injection volume: 10 μL Precursor ion: 605.3 m / z, product ion: 249.0 m / z

[0160] Figure 1 shows in vitro release data from precursor formulations Nos. 32, 33, and 35 containing leuprolide acetate. In Figure 1, the horizontal axis represents time [days], and the vertical axis represents the release rate [%] of leuprolide acetate into PBS solution at each time point relative to the amount of leuprolide acetate contained in the precursor formulation at the start of the test (mean value of n = 3). Precursor formulations Nos. 32, 33, and 35 did not exhibit an initial burst release, demonstrating sustained release of leuprolide acetate. Among these, precursor formulations Nos. 33 and 35 exhibited fast sustained release rates, while precursor formulation No. 32 exhibited a slow sustained release rate.

[0161] The weight ratio of lipid to oil in the precursor formulations Nos. 32, 33, and 35 was the same as that of the precursor formulations Nos. 8, 10, and 12, respectively, and the non-lamellar liquid crystal structure obtained by addition to an aqueous medium was also similar. The liquid crystal phase and unit lattice a [nm] of the gel compositions obtained by adding the precursor formulations Nos. 8, 10, and 12 to water were 7.8 nm for HII, 11 nm for HII, and 20 nm + 11 nm for Pn3m + HII, respectively (from Table 1, the unit lattice a is calculated from the liquid crystal structure and the scattering vector values ​​of peaks including q1). Generally, the more hydrophilic the liquid crystal phase, the faster the sustained release rate (Pn3m > HII > Fd3m), and the larger the unit lattice a, the faster the sustained release rate. Therefore, a sustained release rate in accordance with this rule was obtained.

[0162]

[0163] Example 6 In Vivo Pharmacokinetics Test of Precursor Formulations Containing Leuprolide Acetate Eight-week-old male Wistar rats, whose jugular vein had been cannulated in advance, were given general anesthesia using a triple anesthesia mixture (medetomidine hydrochloride + midazolam + butorphanol tartrate), and after shaving their backs, 100 mg of each of precursor formulations No. 34 and 35 (prepared in Example 5) containing leuprolide acetate was subcutaneously administered to the right back using a syringe equipped with a 23 G needle (Terumo syringe, 1 mL).

[0164] Next, 100 μL of blood was collected from the cannulation tube at 0, 1, 3, and 6 hours and 1, 3, 5, 7, and 10 days after the start of the test. Each blood sample was centrifuged (15,000 rpm, 5 minutes, 4°C) to obtain plasma samples. 50 μL of acetonitrile was added to each of the obtained plasma samples, which were then stirred in a vortex mixer and further centrifuged (15,000 rpm, 5 minutes, 4°C). The resulting supernatants were used as measurement samples.

[0165] The quantification of leuprolide acetate in the measurement sample was carried out by LC / MS / MS analysis in the same manner as in Example 5.

[0166] Figure 2 shows in vitro pharmacokinetic data for precursor formulations No. 34 and 35 containing leuprolide acetate. In Figure 2, the horizontal axis represents time [days], and the vertical axis represents the blood concentration of leuprolide acetate [ng / mL] (mean value of n=5). The AUC (area under the blood concentration-time curve) [ng·h / mL] was calculated from Figure 2 and found to be 2170 for precursor formulation No. 34 and 2152 for precursor formulation No. 35.

[0167] The blood concentration of leuprolide acetate disappeared within 7 days for precursor formulation No. 34, whereas it disappeared within 3 days for precursor formulation No. 35. The weight ratios of lipid and oil in precursor formulations No. 34 and 35 were the same as those in precursor formulations No. 3 and 12, respectively, and the non-lamellar liquid crystal structures obtained by addition to aqueous media were also similar. The liquid crystal phase and unit cell a [nm] of the gel compositions obtained by adding precursor formulations No. 3 and 12 to water were 5.9 nm for HII and 20 nm + 11 nm for Pn3m + HII, respectively (Table 1 shows that unit cell a is calculated from the liquid crystal structure and scattering vector values ​​of peaks including q1). As with the in vitro release test of Example 5, a sustained release rate was obtained that conformed to the rule that the larger the unit cell a, the faster the sustained release rate.

[0168] After 28 days of administration, rats administered with precursor formulation No. 35 were euthanized by exsanguination under inhalation anesthesia with sevoflurane (Mylan Pharmaceuticals), and the administration site, including the skin and subcutaneous tissue, was excised. After simple removal of subcutaneous fat, the administration site was visually observed.

[0169] As shown in the photograph of the administration site in Figure 3, no formulation remained at the administration site, and no findings of inflammation caused by the formulation were macroscopically observed in the surrounding tissue.

[0170] Example 7: Synthesis of amphiphilic compounds (fatty acid esters)

[0171] 2.43 g (31.9 mmol) of propylene glycol in Example 1(4) was mixed with 2.87 g (31.9 mmol) of 1,3-butylene glycol, 3.32 g (31.9 mmol) of 3-methyl-1,3-butanediol, 3.39 g (31.9 mmol) of diethylene glycol, or 4.66 g of isosorbide (31.9 mmol, Sanko Chemical Industry Co., Ltd., purity 98.0% or more, moisture 1.0% or less), or 3.09 g (10.0 mmol) of ethyl linoleate in Example 1(4) and 2.43 g (31.9 mmol) of propylene glycol. The procedure was the same as in Example 1 (4) above, except that 3.11 g (10.0 mmol) of ethyl oleate and 4.66 g (31.9 mmol) of isosorbide were replaced with 3.11 g (10.0 mmol) of ethyl oleate and 4.66 g (31.9 mmol) of isosorbide (when isosorbide was used, 0.3 equivalents of potassium carbonate was used and the reaction was carried out at 95°C), to obtain 1,3-butylene glycol monolinoleate, 3-methyl-1,3-butanediol monolinoleate, diethylene glycol monolinoleate, isosorbide monolinoleate, or isosorbide monooleate as a colorless to pale yellow, transparent liquid. The yield, yield, and １ The H-NMR measurement data was as follows:

[0172] (1) 1,3-butylene glycol monolinoleate

[0173] Yield: 2.67 g (76%) １ H-NMR spectrum (300 MHz, CDCl ３, TMS) δ: 0.89 (m, 3H), 1.2-1.4 (m, 17H), 1.62 (m, 2H), 1.6-1.9 (m, 2H), 1.95-2.1 (m, 4H), 2.30-2.37 (m, 2H), 2.77 (m, 2H), 3.5 -3.7 (m, 0.5H), 3.85 (m, 0.75H), 4.12 (m, 0.75H), 4.35 (ddd, J=5.3, 8.3, 13.6Hz, 0.75H), 5.13 (m, 0.25H), 5.26-5.44 (m, 4H)

[0174] (2) 3-methyl-1,3-butanediol monolinoleate

[0175] Yield: 2.41 g (66%) １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.89 (m, 3H), 1.2-1.4 (m, 20H), 1.62 (m, 2H), 1.84 (t, J=6.9Hz, 2H), 1.95-2.1 (m, 4H), 2.30 (t, J=7.5Hz, 2H), 2.77 (m, 2H), 4.25 (t, J=6.9Hz, 2H), 5.27-5.43 (m, 4H)

[0176] (3) Diethylene glycol monolinoleate

[0177] Yield 2.48 g (67%) １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.89 (m, 3H), 1.2-1.4 (m, 14H), 1.63 (m, 2H), 1.95-2.1 (m, 4H), 2.34 (t, J = 7.6Hz, 2H), 2.77 (m, 2H), 3.61 (m, 2H), 3.70 (m, 2H), 3.74 (m, 2H), 4.25 (m, 2H), 5.27-5.44 (m, 4H)

[0178] (4) Isosorbide monolinoleate

[0179] Yield: 2.31 g (56%) １ H-NMR spectrum (300 MHz, CDCl ３, TMS) δ: 0.89 (m, 3H), 1.2-1.4 (m, 14H), 1.64 (m, 2H), 1.95-2.1 (m, 4H), 2.3-2.4 (m, 2H), 2.77 (m, 2H), 3.55 (dd, J=6.0, 9.5Hz, 0.6H), 3.75 (ddd, J=2.1.5.2, 9.8Hz, 0.4H), 3.8-3.95 (m, 1.8 H), 3.99 (m, 1.2H), 4.2-4.35 (m, 1H), 4.38 (d, J = 4.6Hz, 0.4H), 4.44 (d, J = 4.3Hz, 0.6H), 4.60 ( t, J = 4.9Hz, 0.6H), 4.83 (t, J = 5.0Hz, 0.4H), 5.12 (m, 0.4H), 5.21 (s, 0.6H), 5.26-5.45 (m, 4H)

[0180] (5) Isosorbide monooleate

[0181] Yield 2.38 g (58%) １ H-NMR spectrum (300 MHz, CDCl ３ , TMS) δ: 0.88 (m, 3H), 1.2-1.4 (m, 20H), 1.64 (m, 2H), 1.95-2.15 (m, 4H), 2.25-2.4 (m, 2H) , 3.54 (dd, J=6.0, 9.5Hz, 0.6H), 3.74 (ddd, J=2.1.5.2, 9.8Hz, 0.4H), 3.8-3.95 (m, 1.8H) , 3.99 (m, 1.2H), 4.2-4.35 (m, 1H), 4.37 (d, J = 4.6Hz, 0.4H), 4.44 (d, J = 4.3Hz, 0.6H), 4.6 0 (t, J = 4.9Hz, 0.6H), 4.82 (t, J = 5.0Hz, 0.4H), 5.12 (m, 0.4H), 5.20 (s, 0.6H), 5.34 (m, 2H)

[0182] Example 8 Preparation of Precursor Formulations, Gel Formation Test, and Analysis of Liquid Crystal Structure The amphiphilic compound synthesized in Example 7, SPC (LIPOID S100, Lipoid) as a phospholipid, and EtOH (ethanol) were mixed according to the blending ratios shown in Table 6 below. The resulting mixture was dissolved in a water bath at 40°C or less to prepare precursor formulations No. 36 to 40 shown in Table 6.

[0183] It should be noted that 1,3-butylene glycol monolinoleate, 3-methyl-1,3-butanediol monolinoleate, and isosorbide monolinoleate did not form a gel composition by themselves when mixed with water.

[0184] A gel formation test was conducted on precursor formulations No. 36 to 40. A portion of each precursor formulation (approximately 100 to 300 mg) was added to an excess amount of water for injection (approximately 0.5 to 2 mL) in a vial, and the mixture was mixed at room temperature (25°C) using a spoon and / or a vortex mixer. As a result, for all precursor formulations No. 36 to 40, gel compositions that were colorless, transparent, or opaque in appearance were obtained that separated into the excess water for injection (aqueous medium), and these were thought to exhibit a non-lamellar liquid crystal structure consisting of reverse hexagonal liquid crystals or cubic liquid crystals.

[0185]

[0186] The upper and / or lower limits of the numerical ranges described herein can be arbitrarily combined to define a preferred range. For example, the upper and lower limits of the numerical ranges can be arbitrarily combined to define a preferred range, the upper limits of the numerical ranges can be arbitrarily combined to define a preferred range, and the lower limits of the numerical ranges can be arbitrarily combined to define a preferred range.

[0187] Although the present embodiment has been described in detail above, the specific configuration is not limited to this embodiment, and even if there are design changes within the scope that do not deviate from the gist of this disclosure, they are included in this disclosure.

[0188] Furthermore, all publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A non-lamellar liquid crystal-forming composition comprising a fatty acid ester and a phospholipid, wherein the fatty acid ester has a hydrophilic group having one hydroxyl group or a salt thereof, and the fatty acid ester is a fatty acid ester that does not have any of the structures represented by the following general formulas (A) to (D). 【Chemical 1】 (In general formulas (A) to (D), m is independently 1 or 2, a, b, c, and d each independently represent a bonding site with the hydrophilic group, 【Chemical Formula 2】 each independently represents a single bond or a double bond, n in general formula (A) is 0, 1, or 2, and the wavy lines in general formulas (B), (C), and (D) each independently represent the E isomer or the Z isomer.)

2. The composition according to claim 1, wherein the fatty acid of the fatty acid ester has 6 to 24 carbon atoms.

3. The composition according to claim 1, wherein the weight ratio of the fatty acid ester to the phospholipid is 90:10 to 20:

80.

4. The composition according to claim 1, wherein the fatty acid ester is a fatty acid ester of at least one selected from saturated or unsaturated straight-chain fatty acids and their derivatives and at least one selected from glycols and their derivatives.

5. The composition according to claim 4, wherein the straight-chain fatty acid is a straight-chain fatty acid having an unsaturation degree of 0 to 6.

6. The composition according to claim 4, wherein the straight-chain fatty acid is a straight-chain fatty acid represented by the following general formula (I). 【Chemical Formula 3】 (In general formula (I), R is a linear hydrocarbon group represented by -C ｐ H ｑ -, p is an integer from 4 to 22, q is an integer from 2p - 4 to 2p when p is 5 or less, q is an integer from 2p - 6 to 2p when p is 6 or 7, q is an integer from 2p - 8 to 2p when p is 8 or 9, q is an integer from 2p - 10 to 2p when p is 10 or 11, and q is an integer from 2p - 12 to 2p when p is 12 or more.)

7. The composition according to claim 4, wherein the straight-chain fatty acid is at least one fatty acid selected from caprylic acid, lauric acid, myristic acid, palmitic acid, oleic acid, linoleic acid, and behenic acid.

8. The composition according to claim 4, wherein the glycol is at least one glycol selected from propylene glycol, ethylene glycol, butylene glycol, 3-methyl-1,3-butanediol, diethylene glycol, and isosorbide.

9. The composition according to claim 1 or 4, wherein the phospholipid is at least one phospholipid selected from phosphatidylcholine, phosphatidylethanolamine, and their salts.

10. The composition according to claim 1 or 4, wherein the phospholipid is at least one phospholipid selected from soybean phosphatidylcholine, egg yolk phosphatidylcholine, dimyristoyl phosphatidylcholine, dioleyl phosphatidylcholine, dioleyl phosphatidylethanolamine, and salts thereof.

11. The composition according to claim 1 or 4, comprising at least one of an oil component and an organic solvent.

12. The composition according to claim 1 or 4, which is a non-lamellar liquid crystal composition further comprising an aqueous medium.

13. The composition according to claim 12, which is a non-lamellar liquid crystal emulsion composition further comprising a surfactant.

14. The composition according to claim 1 or 4, which is a liquid crystal precursor composition capable of forming a non-lamellar liquid crystal in the presence of an aqueous medium.

15. A pharmaceutical preparation comprising the composition according to claim 1 or 4.

16. The pharmaceutical preparation according to claim 15, for preventing adhesion of biological tissues.

17. The pharmaceutical preparation according to claim 15, which is a sustained-release preparation, wherein the composition further comprises a drug.

18. The pharmaceutical preparation according to claim 17, wherein the drug is a gonadotropin-releasing hormone (GnRH) agonist.

19. The pharmaceutical preparation according to claim 18, wherein the GnRH agonist is leuprorelin or a salt thereof.

20. The pharmaceutical preparation according to claim 15, which is a spray, an aerosol, an injection, or a depot preparation.

21. A compound represented by formula (X') or a salt thereof. 【Chemical Formula 4】 (In general formula (X'), R １ is a structure derived from propylene glycol, butylene glycol, isoprene glycol, diethylene glycol or isosorbide.)

22. A compound represented by formula (X), (Y), or (Z) or a salt thereof. 【Chemical Formula 5】

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