Non-lamellar liquid crystal-forming composition with high safety

Abstract

The present application provides a non-lamellar liquid crystal forming composition with high safety. The present application relates to a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by the following general formula (I) and a phospholipid, the phospholipid improving the biocompatibility of the non-lamellar liquid crystal forming composition; in the formula, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2, m represents 1 or 2, and R represents a hydrophilic group having one or more hydroxyl groups.

Description

TECHNICAL FIELD

[0001] The present application relates to a non-lamellar liquid crystal-forming composition with high safety. BACKGROUND

[0002] Liposomes and other lyotropic liquid crystals have been reported to be useful as biomimetic drug delivery system (DDS) carriers since the period when the concept of DDS was advocated. In recent years, non-lamellar liquid crystals (NLLCs), which are one type of lyotropic liquid crystals, have been reported to have advantages such as high drug content and ease of preparation compared to existing DDS carriers.

[0003] Various liquid crystal-forming compounds are used for various purposes in the fields of cosmetics, pharmaceuticals, and the like. In recent years, amphiphilic compounds having isoprenoid-type fatty chains that can form cubic liquid crystals exhibiting high stability even at low temperatures (below 6°C) have been developed (Patent Literature 1). Amphiphilic compounds having isoprenoid-type fatty chains that are low in viscosity and can form non-lamellar liquid crystals and be used as bases for injectable agents have also been developed (Patent Literature 2). Patent Literature 3 reports that amphiphilic compounds having isoprenoid-type fatty chains that can form non-lamellar liquid crystals can be used as antiblocking agents.

[0004] On the other hand, glycerol monooleate (GMO) and phytantriol (PHY), which are liquid crystal-forming compounds (self-organizing lipids / SOL) used for food additives and skin and hair cosmetics, are considered to be safe in terms of external application to humans and oral ingestion, but have been pointed out to have serious problems related to safety such as cytotoxicity and hemolyticity in terms of in vivo use (for example, Non-Patent Literature 1 and Non-Patent Literature 2).

[0005] However, a general technique capable of improving safety in in vivo application while maintaining the effectiveness of pharmaceutical preparations containing liquid crystal-forming compounds has not yet been established.

[0006] Prior Art Documents

[0007] Patent Literature

[0008] Patent Literature 1: International Publication No. 2006 / 043705

[0009] Patent Literature 2: International Publication No. 2011 / 078383

[0010] Patent Literature 3: International Publication No. 2014 / 178256

[0011] Non-Patent Literature

[0012] Non-patent literature 1: Hinton T.M., et al., Toxicology Research, Vol. 3, (2014) p. 11-22

[0013] Non-patent literature 2: Wibroe P.P., et al., Nanomedicine: Nanotechnology, Biology and Medicine, Vol. 11 (2015) p. 1909-1914 SUMMARY

[0014] PROBLEMS TO BE SOLVED BY THE INVENTION

[0015] The present application has an object to provide a non-lamellar liquid crystal forming composition with high safety. The present application has an object to provide a non-lamellar liquid crystal forming composition using a novel amphiphilic compound.

[0016] TECHNICAL SOLUTION FOR SOLVING THE PROBLEMS

[0017] In order to solve the above problems, the present inventors have conducted intensive studies, and as a result, have found that a non-lamellar liquid crystal forming composition with high safety allowing in vivo application, and improved biocompatibility can be produced by adding a phospholipid to an amphiphilic compound having a specific isoprenoid-type aliphatic chain. The present inventors have also successfully synthesized a novel amphiphilic compound having an isoprenoid-type aliphatic chain, which can be used for producing a non-lamellar liquid crystal forming composition. Thus, the present inventors have completed the present application.

[0018] That is, the present application includes the following.

[0019] [1] A non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by the following general formula (I) and a phospholipid that improves the biocompatibility of the non-lamellar liquid crystal forming composition,

[0020] [Chemical Formula 1]

[0021]

[0022] In the formula, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2, m represents 1 or 2, represents a single bond or a double bond, and R represents a hydrophilic group having one or more hydroxyl groups.

[0023] [2] The composition according to the above [1], wherein the amphiphilic compound is represented by the following general formula (III) or (IV):

[0024] [Chemical Formula 2]

[0025]

[0026] wherein X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2, and m represents 1 or 2.

[0027] [3] The composition according to any one of the above [1] or [2], wherein n = 2 and m = 2 in the general formula.

[0028] [4] The composition according to any one of the above [1] to [3], wherein the weight ratio of the amphiphilic compound to the phospholipid is 80:20 to 20:80, or 70:30 to 30:70.

[0029] [5] The composition according to the above [4], wherein the weight ratio of the amphiphilic compound to the phospholipid is 50:50 to 30:70, or 45:55 to 30:70.

[0030] [6] The composition according to any one of the above [1] to [5], wherein the phospholipid is phosphatidylcholine or phosphatidylethanolamine.

[0031] [7] The composition according to any one of the above [1] to [6], wherein the phospholipid is selected from the group consisting of soybean phosphatidylcholine, egg yolk phosphatidylcholine, dimyristoyl phosphatidylcholine, dioleoyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine.

[0032] [8] The composition according to any one of the above [1] to [7], wherein R in the general formula (I) represents a hydrophilic group obtained by removing one hydroxyl group from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, and glycol.

[0033] [9] The composition according to the above [1] or [2], wherein the amphiphilic compound is selected from the group consisting of:

[0034] mono-O-(5,9,13,17-tetramethyl octadeca-4-enoyl) glycerol;

[0035] mono-O-(5,9,13,17-tetramethyl octadecanoyl) glycerol;

[0036] mono-O-(5,9,13,17-tetramethyl octadeca-4,8,12,16-tetraenoyl) glycerol;

[0037] mono-O-(5,9,13,17-tetramethyl octadecanoyl) erythritol;

[0038] mono-O-(5,9,13,17-tetramethyl octadeca-4-enoyl) pentaerythritol;

[0039] Mono-O-(5,9,13,17-tetramethylheptadecanoyl) pentaerythritol;

[0040] Mono-O-(5,9,13,17-tetramethylheptadecan-4-enoyl) diglycerol;

[0041] Mono-O-(5,9,13,17-tetramethylheptadecan-4-enoyl) sorbitan;

[0042] Mono-O-(5,9,13,17-tetramethylheptadecanoyl) sorbitan;

[0043] Mono-O-(5,9,13,17-tetramethylheptadecan-4-enoyl) isosorbide;

[0044] Mono-O-(5,9,13,17-tetramethylheptadecanoyl) isosorbide;

[0045] Mono-O-(5,9,13-trimethyltetradecan-4-enoyl) glycerol;

[0046] Mono-O-(5,9,13-trimethyltetradecan-4-enoyl) sorbitan;

[0047] Mono-O-(5,9,13-trimethyltetradecanoyl) sorbitan;

[0048] Mono-O-(5,9,13-trimethyltetradecan-4-enoyl) propylene glycol;

[0049] Mono-O-(5,9,13,17-tetramethylheptadecan-4-enoyl) propylene glycol;

[0050] Mono-O-(5,9,13,17-tetramethylheptadecanoyl) propylene glycol;

[0051] Mono-O-(4,8,12,16-tetramethylheptadecan-3-enoyl) sorbitan;

[0052] Mono-O-(4,8,12,16-tetramethylheptadecan-3-enoyl) isosorbide;

[0053] Mono-O-(4,8,12,16-tetramethylheptadecan-3-enoyl) propylene glycol;

[0054] Mono-O-(5,9,13,17-tetramethylheptadecan-4-enoyl) ethylene glycol;

[0055] Mono-O-(5,9,13,17-tetramethylheptadecan-4-enoyl) 1,3-butanediol; and,

[0056] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) 3-methyl-1,3-butanediol.

[0057]

[10] The composition according to any one of the above [1] to [9], wherein the improvement of biocompatibility is manifested by a decrease or disappearance of toxicity compared to a control composition not containing a phospholipid.

[0058]

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

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

[0059]

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

[11] , wherein the composition is a non-lamellar liquid crystal composition further containing an aqueous medium.

[0060]

[13] The composition according to the above

[12] , wherein the composition is a non-lamellar liquid crystal emulsion composition further containing a surfactant.

[0061]

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

[11] , wherein the composition is a liquid crystal precursor composition not containing an aqueous medium and capable of forming a non-lamellar liquid crystal in the presence of an aqueous medium.

[0062]

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

[14] .

[0063]

[16] The pharmaceutical preparation according to the above

[15] , which is used for preventing adhesion of biological tissues.

[0064]

[17] The pharmaceutical preparation according to the above

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

[0065]

[18] The pharmaceutical preparation according to the above

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

[0066]

[19] The pharmaceutical preparation according to the above

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

[0067]

[20] The pharmaceutical preparation according to any one of the above

[15] to

[19] , which is a spray, an aerosol, an injection, or a long-acting agent.

[0068]

[21] An amphiphilic compound represented by the following general formula (I’):

[0069] [Chemical 3]

[0070]

[0071] In the formula, X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2, m represents 1 or 2, represents a single bond or a double bond, and R represents a hydrophilic group obtained by removing one hydroxyl group from any one selected from the group consisting of sorbitan, isosorbide, and a diol.

[0072]

[22] The compound or salt thereof according to the above

[21] , wherein, in the general formula (I'), n = 2.

[0073]

[23] The compound or salt thereof according to the above

[21] or

[22] , wherein the amphiphilic compound is selected from the group consisting of:

[0074] mono O-(5,9,13,17-tetramethyl octadeca-4-enoyl) sorbitan;

[0075] mono O-(5,9,13,17-tetramethyl octadecanoyl) sorbitan;

[0076] mono O-(5,9,13-trimethyl tetradecan-4-enoyl) sorbitan;

[0077] mono O-(5,9,13-trimethyl tetradecanoyl) sorbitan;

[0078] mono O-(5,9,13,17-tetramethyl octadeca-4-enoyl) isosorbide;

[0079] mono O-(5,9,13,17-tetramethyl octadecanoyl) isosorbide;

[0080] mono O-(5,9,13-trimethyl tetradecan-4-enoyl) propylene glycol;

[0081] mono O-(5,9,13,17-tetramethyl octadeca-4-enoyl) propylene glycol;

[0082] mono O-(5,9,13,17-tetramethyl octadecanoyl) propylene glycol;

[0083] mono O-(4,8,12,16-tetramethyl heptadeca-3-enoyl) sorbitan;

[0084] mono O-(4,8,12,16-tetramethyl heptadeca-3-enoyl) isosorbide;

[0085] mono O-(4,8,12,16-tetramethyl heptadeca-3-enoyl) propylene glycol;

[0086] mono O-(5,9,13,17-tetramethyl octadeca-4-enoyl) ethylene glycol;

[0087] mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) 1,3-butanediol; and

[0088] mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) 3-methyl-1,3-butanediol.

[0089] This specification includes the disclosure of Japanese Patent Application Nos. 2020-011237 and 2020-211758, which are the basis for priority of this application.

[0090] Effects of the Invention

[0091] The present application can provide a non-lamellar liquid crystal-forming composition with high safety. The present application can also provide a non-lamellar liquid crystal-forming composition using a novel amphiphilic compound. BRIEF DESCRIPTION OF DRAWINGS

[0093] Figure 1 is a photograph showing the appearance of an abscess produced due to administration of a reagent. Figure 1 A to C: precursor formulations of No. 1 formulation. Figure 1 D: precursor formulation of No. 71 formulation. A is an abscess produced along the incision and sutured wound, B is a part of the abscess taken out from the inside, C is the site of abscess production after the abscess (part) is taken out, and D is an abscess produced along the incision and sutured wound. The arrow indicates the abscess (part).

[0094] Figure 2 shows the release rate of leuprolide acetate from each precursor formulation. As a phospholipid, No. 121 precursor formulation contains DMPC, No. 122 precursor formulation contains DOPC, No. 123 precursor formulation contains DOPE, and No. 124 precursor formulation contains DOPG-Na. No. 125 aqueous solution contains neither C17 glyceride nor phospholipid.

[0095] Figure 3 is a photograph showing the results of subcutaneous implantation test using a precursor formulation containing DMPC (A) or DOPC (B) as a phospholipid. Figure 3 A) or DOPC ( Figure 3 B) as a phospholipid.

[0096] Figure 4 shows the release rate of FD-4 from each precursor formulation. The values represent the mean ± S.D. (n = 3).

[0097] Figure 5 shows the release rate of leuprolide acetate from each precursor formulation. The values represent the mean ± S.D. (n = 3).

[0098] Figure 6is a photograph showing the result of a subcutaneous implant test using an emulsion or a precursor formulation containing a isoprenoid-type lipid and a phospholipid.

[0099] Figure 7 is a photograph showing the result of a subcutaneous implant test using a precursor formulation containing a isoprenoid-type lipid and a phospholipid. DETAILED DESCRIPTION

[0100] The present application will be described in detail below.

[0101] The present application relates to a composition containing an amphiphilic compound having a specific isoprenoid-type fatty chain, particularly a non-lamellar liquid crystal-forming composition. In one embodiment, the present application relates to a non-lamellar liquid crystal-forming composition containing an amphiphilic compound having a specific isoprenoid-type fatty chain and a phospholipid. The non-lamellar liquid crystal-forming composition according to the present application is particularly preferably a non-lamellar liquid crystal-forming composition having improved biocompatibility by a phospholipid. Such a non-lamellar liquid crystal-forming composition is high in safety, and thus allows application in vivo.

[0102] In the present application, an amphiphilic compound (isoprenoid-type lipid) having an isoprenoid-type fatty chain represented by the following general formula (I) can be combined with a phospholipid to produce a non-lamellar liquid crystal-forming composition. The present application provides a composition containing an amphiphilic compound represented by the general formula (I) and a phospholipid, particularly a non-lamellar liquid crystal-forming composition.

[0103] [Chemical Formula 4]

[0104]

[0105] In one embodiment, in the general formula (I), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2 (n = 1 or 2), and m represents 1 or 2 (m = 1 or 2). The combination of n and m can be any one of n = 1, m = 1; n = 1, m = 2; n = 2, m = 1; or n = 2, m = 2; preferably the combination of n = 2, m = 1 and n = 2, m = 2, and more preferably the combination of n = 2, m = 2. In one embodiment, in the general formula (I), X and Y together represent an oxygen atom, and n = 2, m = 2 or n = 2, m = 1.

[0106] In another embodiment, in the general formula (I) and the following general formulae (II) to (IV), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, and m represents 1 or 2.

[0107] In addition, in the present application, in the chemical formula of the amphiphilic compound, represents a single bond or a double bond.

[0108] R in General Formula (I) represents a hydrophilic group having 1 or 2 or more (for example, 2, 3, 4, or 5) hydroxyl groups. R in General Formula (I) can also be a residue after removing 1 hydroxyl group (OH) from a polyol. R in General Formula (I) can also have an ether bond and / or a cyclic structure. R in General Formula (I) can be exemplified by, but not limited to, a hydrophilic group after removing 1 hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglyceride, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, xylitol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and other butanediols, and isopropylidene glycol). In addition, isopropylidene glycol can also be referred to as 3-methyl-1,3-butanediol. R in General Formula (I) is more preferably a hydrophilic group after removing 1 hydroxyl group (OH) from glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, or a diol (for example, propylene glycol, ethylene glycol, 1,3-butanediol and other butanediols, or isopropylidene glycol). In one embodiment, R in General Formula (I) can be, for example, a hydrophilic group having 1 hydroxyl group after removing 1 hydroxyl group (OH) from a diol having no ether bond, and such an amphiphilic compound represented by General Formula (I) is not a self-organizing lipid (SOL).

[0109] In addition, with respect to the present application, in the chemical formula representing the amphiphilic compound, means that the amphiphilic compound is an E isomer (cis isomer) or a Z isomer (trans isomer) of geometric isomers or a mixture thereof.

[0110] As an example of the amphiphilic compound represented by General Formula (I), an amphiphilic compound represented by the following General Formula (II) can be exemplified.

[0111] [Chemical 5]

[0112]

[0113] In General Formula (II), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2 (n = 1 or 2), and m represents 1 or 2 (m = 1 or 2). The combination of n and m can be any one of n = 1, m = 1; n = 1, m = 2; n = 2, m = 1; or n = 2, m = 2; preferably the combination of n = 2, m = 1 and n = 2, m = 2, and more preferably the combination of n = 2, m = 2. In one embodiment, in General Formula (II), X and Y together represent an oxygen atom, and n = 2, m = 2 or n = 2, m = 1.

[0114] R in General Formula (II) represents a hydrophilic group having 1 or more hydroxyl groups. R in General Formula (II) can be the same as R in General Formula (I). R in General Formula (II) can be, for example, but not limited to, a hydrophilic group obtained by removing 1 hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglyceride, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, xylitol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and other butanediols, and isopentyl glycol). R in General Formula (II) is more preferably a hydrophilic group obtained by removing 1 hydroxyl group (OH) from glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, or a diol (for example, propylene glycol, ethylene glycol, 1,3-butanediol and other butanediols, or isopentyl glycol and the like). In one embodiment, R in General Formula (II) can be, for example, a hydrophilic group having 1 hydroxyl group obtained by removing 1 hydroxyl group (OH) from a diol having no ether bond.

[0115] As another example of the amphiphilic compound represented by General Formula (I), an amphiphilic compound represented by General Formula (III) below can be given.

[0116] [Chemical 6]

[0117]

[0118] In General Formula (III), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2 (n = 1 or 2), and m represents 1 or 2 (m = 1 or 2). The combination of n and m can be any one of n = 1, m = 1; n = 1, m = 2; n = 2, m = 1; or n = 2, m = 2; preferably the combination of n = 2, m = 1 and n = 2, m = 2, and more preferably the combination of n = 2, m = 2. In one embodiment, in General Formula (III), X and Y together represent an oxygen atom, and n = 2, m = 2 or n = 2, m = 1.

[0119] R in General Formula (III) represents a hydrophilic group having 1 or more hydroxyl groups. R in General Formula (III) can be the same as R in General Formula (I). R in General Formula (III) can be, for example, but not limited to, a hydrophilic group obtained by removing 1 hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglyceride, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, xylitol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and other butanediols, and isopentyl glycol). R in General Formula (III) is more preferably a hydrophilic group obtained by removing 1 hydroxyl group (OH) from glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, or a diol (for example, propylene glycol, ethylene glycol, 1,3-butanediol and other butanediols, or isopentyl glycol and the like). In one embodiment, R in General Formula (III) can be, for example, a hydrophilic group having 1 hydroxyl group obtained by removing 1 hydroxyl group (OH) from a diol having no ether bond.

[0120] As still another example of the amphiphilic compound represented by General Formula (I), an amphiphilic compound represented by General Formula (IV) below can be given.

[0121] [Chemical 7]

[0122]

[0123] In General Formula (IV), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2 (n = 1 or 2), and m represents 1 or 2 (m = 1 or 2). The combination of n and m can be any one of n = 1, m = 1; n = 1, m = 2; n = 2, m = 1; or n = 2, m = 2; preferably the combination of n = 2, m = 1 and n = 2, m = 2, and more preferably the combination of n = 2, m = 2. In one embodiment, in General Formula (IV), X and Y together represent an oxygen atom, and n = 2, m = 2 or n = 2, m = 1.

[0124] R in general formula (IV) represents a hydrophilic group having one or more hydroxyl groups. R in general formula (IV) can be the same as R in general formula (I). R in general formula (IV) can be, for example, but not limited to, a hydrophilic group obtained by removing one hydroxyl group (OH) from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, glyceric acid, triglyceride, xylose, sorbitol, ascorbic acid, glucose, galactose, mannose, dipentaerythritol, maltose, mannitol, xylitol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and the like butanediol, and isopentyl glycol). R in general formula (IV) is more preferably a hydrophilic group obtained by removing one hydroxyl group (OH) from glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, or a diol (for example, propylene glycol, ethylene glycol, 1,3-butanediol and the like butanediol, or isopentyl glycol and the like). In one embodiment, R in general formula (IV) can be, for example, a hydrophilic group having one hydroxyl group obtained by removing one hydroxyl group (OH) from a diol having no ether bond.

[0125] As a preferred example of the amphiphilic compound represented by general formula (I) used in the present application, the following compounds can be given, but are not limited thereto:

[0126] mono-O-(5,9,13,17-tetramethyl octadeca-4-eneacyl) glycerol;

[0127] mono-O-(5,9,13,17-tetramethyl octadecanoyl) glycerol;

[0128] mono-O-(5,9,13,17-tetramethyl octadeca-4,8,12,16-tetraeneacyl) glycerol;

[0129] mono-O-(5,9,13,17-tetramethyl octadecanoyl) erythritol;

[0130] mono-O-(5,9,13,17-tetramethyl octadeca-4-eneacyl) pentaerythritol;

[0131] mono-O-(5,9,13,17-tetramethyl octadecanoyl) pentaerythritol;

[0132] mono-O-(5,9,13,17-tetramethyl octadeca-4-eneacyl) diglycerol;

[0133] mono-O-(5,9,13,17-tetramethyl octadeca-4-eneacyl) sorbitan;

[0134] mono-O-(5,9,13,17-tetramethyl octadecanoyl) sorbitan;

[0135] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) glycerol;

[0136] Mono-O-(5,9,13,17-tetramethyloctadecanoyl) glycerol;

[0137] Mono-O-(5,9,13-trimethyltetradec-4-enoyl) glycerol;

[0138] Mono-O-(5,9,13-trimethyltetradec-4-enoyl) sorbitol;

[0139] Mono-O-(5,9,13-trimethyltetradecanoyl) sorbitol;

[0140] Mono-O-(5,9,13-trimethyltetradec-4-enoyl) propylene glycol;

[0141] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) propylene glycol;

[0142] Mono-O-(5,9,13,17-tetramethyloctadecanoyl) propylene glycol;

[0143] Mono-O-(4,8,12,16-tetramethylheptadec-3-enoyl) sorbitol;

[0144] Mono-O-(4,8,12,16-tetramethylheptadec-3-enoyl) isosorbide;

[0145] Mono-O-(4,8,12,16-tetramethylheptadec-3-enoyl) propylene glycol;

[0146] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) ethylene glycol;

[0147] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) 1,3-butanediol; and,

[0148] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) 3-methyl-1,3-butanediol.

[0149] As another example of the amphiphilic compound represented by General Formula (I), the following compounds can also be given, but are not limited thereto:

[0150] Mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl) sorbitol;

[0151] Mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl) isosorbide;

[0152] mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol;

[0153] mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol;

[0154] mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol.

[0155] In the preferred embodiment, the amphiphilic compound represented by the general formula (I) used in the present application can be a compound that exhibits low viscosity by itself. Specifically, as for the viscosity possessed by the amphiphilic compound represented by the general formula (I) by itself, the measured value at 25°C is preferably 15.0 Pa-s or less, more preferably 11.0 Pa-s or less, and further preferably 6.0 Pa-s or less. This viscosity can be measured, for example, using a viscoelasticity measuring device (Gemini II, Malvern Instruments Ltd.) at a temperature of 25°C.

[0156] The non-lamellar liquid crystal-forming composition according to the present application can also contain one or more than two kinds of the amphiphilic compound having a fatty chain of isoprenoid type represented by the general formula (I). When the non-lamellar liquid crystal-forming composition according to the present application contains a plurality of kinds of the amphiphilic compound having a fatty chain of isoprenoid type, the weight ratio of these amphiphilic compounds is not particularly limited. However, in one embodiment, the weight ratio of mono-O-(5,9,13-trimethyltetradecanoyl) sorbitol and mono-O-(5,9,13-trimethyltetradecanoyl) isosorbide contained in the non-lamellar liquid crystal-forming composition according to the present application can be in the range of 50:50 to 99:1; preferably in the range of 60:40 to 90:10, 70:30 to 90:10, or 80:20 to 90:10, for example, in the range of 8:2 or 85:15. In one embodiment, the weight ratio of mono-O-(5,9,13,17-tetramethyl octadecanoyl) sorbitol and mono-O-(5,9,13,17-tetramethyl octadecanoyl) isosorbide contained in the non-lamellar liquid crystal-forming composition according to the present application can be in the range of 50:50 to 99:1; preferably in the range of 60:40 to 90:10, 70:30 to 90:10, or 80:20 to 90:10, for example, in the range of 8:2 or 85:15. In one embodiment, the weight ratio of mono-O-(5,9,13,17-tetramethyl octadecanoyl) sorbitol and mono-O-(5,9,13,17-tetramethyl octadecanoyl) isosorbide contained in the non-lamellar liquid crystal-forming composition according to the present application can be in the range of 50:50 to 99:1; preferably in the range of 60:40 to 90:10, 70:30 to 90:10, or 80:20 to 90:10, for example, in the range of 8:2 or 85:15. In one embodiment, the weight ratio of mono-O-(5,9,13,17-tetramethyl octadecanoyl) sorbitol and mono-O-(5,9,13,17-tetramethyl octadecanoyl) isosorbide contained in the non-lamellar liquid crystal-forming composition according to the present application can be in the range of 50:50 to 99:1; preferably in the range of 60:40 to 90:10, 70:30 to 90:10, or 80:20 to 90:10, for example, in the range of 8:2 or 85:15. In one embodiment, the weight ratio of 2-O-(5,9,13,17-tetramethyl octadecanoyl) isosorbide and 5-O-(5,9,13,17-tetramethyl octadecanoyl) isosorbide contained in the non-lamellar liquid crystal-forming composition according to the present application can be in the range of 50:50 to 99:1; preferably in the range of 50:50 to 70:30 or 55:45 to 65:35, for example, in the range of 60:40.In one embodiment, the weight ratio of mono-O-(4,8,12,16-tetramethylheptadec-3-enoyl) sorbitan and mono-O-(4,8,12,16-tetramethylheptadec-3-enoyl) isosorbide contained in the non-lamellar liquid crystal forming composition according to the present application can be in the ratio of 50:50 to 99:1; preferably in the ratio of 60:40 to 90:10, 70:30 to 90:10, or 80:20 to 90:10, for example, 8:2 or 85:15. In one embodiment, the weight ratio of mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl) sorbitan and mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl) isosorbide contained in the non-lamellar liquid crystal forming composition can be in the ratio of 50:50 to 99:1; preferably in the ratio of 60:40 to 90:10, 70:30 to 90:10, or 80:20 to 90:10, for example, 8:2 or 85:15.

[0157] The present application also provides a newly discovered amphiphilic compound represented by the following general formula (I') within the scope of the amphiphilic compounds represented by general formula (I). The amphiphilic compound represented by general formula (I') can also be combined with phospholipids and is preferably used for the production of a non-lamellar liquid crystal forming composition.

[0158] [Chemical Formula 8]

[0159]

[0160] In one embodiment, in general formula (I'), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents 1 or 2, preferably n = 2, and m represents 1 or 2.

[0161] represents a single bond or a double bond, and R represents a hydrophilic group from which one hydroxyl group is removed from sorbitan, isosorbide, or a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol, etc. butanediol, isopropylidene glycol).

[0162] In general formula (I'), the combination of n and m can be any one of n = 1, m = 1; n = 1, m = 2; n = 2, m = 1; or n = 2, m = 2. In one embodiment, in general formula (I'), X and Y together represent an oxygen atom, and n = 2, m = 2, or n = 2, m = 1.

[0163] In another embodiment, in general formula (I'), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2, and m represents 1 or 2,

[0164] represents a single or double bond, R can represent a hydrophilic group after removing one hydroxyl group from: sorbitan, isosorbide, or a diol (e.g., propylene glycol, etc.). The combination of n and m can be any one of n = 0, m = 1; n = 0, m = 2; n = 1, m = 1; n = 1, m = 2; n = 2, m = 1; or n = 2, m = 2. In one embodiment, in general formula (I'), X and Y together represent an oxygen atom, and n = 2, m = 2, or n = 2, m = 1.

[0165] The amphiphilic compound represented by general formula (I') is a compound represented by the above general formulae (II), (III), or (IV), X and Y each represent a hydrogen atom or together represent an oxygen atom, n represents an integer of 0 to 2 (preferably, n = 1 or 2), m represents 1 or 2, and R can represent a hydrophilic group after removing one hydroxyl group from: sorbitan, isosorbide, or a diol (e.g., propylene glycol, etc.).

[0166] As preferred examples of the amphiphilic compound represented by general formula (I'), the following compounds can be given, but are not limited thereto:

[0167] mono-O-(5, 9, 13, 17-tetramethyl octadeca-4-enoyl) sorbitan;

[0168] mono-O-(5, 9, 13, 17-tetramethyl octadecanoyl) sorbitan;

[0169] mono-O-(5, 9, 13-trimethyl tetradecanoyl) sorbitan;

[0170] mono-O-(5, 9, 13-trimethyl tetradecanoyl) sorbitan;

[0171] mono-O-(5, 9, 13, 17-tetramethyl octadeca-4-enoyl) isosorbide;

[0172] mono-O-(5, 9, 13, 17-tetramethyl octadecanoyl) isosorbide;

[0173] mono-O-(5, 9, 13-trimethyl tetradecanoyl) propylene glycol;

[0174] mono-O-(5, 9, 13, 17-tetramethyl octadeca-4-enoyl) propylene glycol;

[0175] mono-O-(5, 9, 13, 17-tetramethyl octadecanoyl) propylene glycol;

[0176] mono-O-(4, 8, 12, 16-tetramethyl heptadeca-3-enoyl) sorbitan;

[0177] Mono O-(4, 8, 12, 16-tetramethylheptadec-3-enoyl) erythritol;

[0178] Mono O-(4, 8, 12, 16-tetramethylheptadec-3-enoyl) erythritol;

[0179] Mono O-(5, 9, 13, 17-tetramethyl octadec-4-enoyl) 1, 3-butanediol; and,

[0180] Mono O-(5, 9, 13, 17-tetramethyl octadec-4-enoyl) 1, 3-butanediol; and,

[0181] Mono O-(5, 9, 13, 17-tetramethyl octadec-4-enoyl) 1, 3-butanediol; and,

[0182] The non-lamellar liquid crystal-forming composition according to the present application contains an amphiphilic compound represented by the general formula (I). The amphiphilic compound represented by the general formula (I) includes an amphiphilic compound represented by the general formula (I'). The non-lamellar liquid crystal-forming composition according to the present application can also contain the above-mentioned amphiphilic compound represented by the general formula (I) or (I') in the form of a salt. The salt of the amphiphilic compound represented by the general formula (I) or the general formula (I') according to the present application can be any salt, and examples thereof include, but are not limited to, salts of alkali metals such as sodium, potassium, calcium, magnesium, and the like, or salts of alkaline earth metals, and the like. The salt of the amphiphilic compound represented by the general formula (I) or the general formula (I') according to the present application can be a pharmaceutically acceptable salt. The non-lamellar liquid crystal-forming composition according to the present application containing the amphiphilic compound represented by the general formula (I) or the general formula (I') in the form of a salt according to the present application is also included in the scope of the non-lamellar liquid crystal-forming composition containing the amphiphilic compound represented by the general formula (I) or the general formula (I') according to the present application.

[0183] The amphiphilic compound represented by the general formula (I) in the present application can be used in combination with a phospholipid. When the amphiphilic compound of the general formula (I) has toxicity, the phospholipid can reduce the toxicity of the amphiphilic compound, improve the biocompatibility of the non-lamellar liquid crystal-forming composition, and improve the safety when the composition is used in vivo. That is, the non-lamellar liquid crystal-forming composition containing the amphiphilic compound represented by the general formula (I) and the phospholipid according to the present application is preferably a non-lamellar liquid crystal-forming composition whose biocompatibility is improved by the phospholipid, as compared with a control composition not containing the phospholipid. In the present application, "biocompatibility" means a property that is not likely to cause or does not cause harmful reactions (side effects) in vivo when used in vivo. The improvement of the biocompatibility in the non-lamellar liquid crystal-forming composition containing the amphiphilic compound represented by the general formula (I) and the phospholipid according to the present application can be confirmed by the reduction or disappearance of toxicity when the composition is used in vivo, as compared with a control composition not containing the phospholipid. In the present application, the "control composition not containing the phospholipid" means a non-lamellar liquid crystal-forming composition having the same composition except that it does not contain the phospholipid. As the "toxicity" related to the present application, the following toxicities can be given, but are not limited to: systemic toxicity such as hepatotoxicity; local toxicity such as generation of foreign body reactions and bleeding, and tissue disorders such as discoloration; and the like. The hepatotoxicity can be confirmed, for example, by the generation of at least one symptom indicating liver disorder selected from the following symptoms: ascites, hepatomegaly, adhesion around the liver (particularly, adhesion at a non-injured and non-inflamed site), liver whitening, and the like. In one embodiment, in a small experimental animal represented by a mouse and a rat, the non-lamellar liquid crystal-forming composition of the present application has reduced or disappeared toxicity as compared with a control composition not containing the phospholipid, so that the mortality after the non-lamellar liquid crystal-forming composition is used in vivo is reduced. The non-lamellar liquid crystal-forming composition containing the amphiphilic compound represented by the general formula (I) and the phospholipid according to the present application allows in vivo use (preferably, non-oral administration such as intraperitoneal administration, intramuscular administration, subcutaneous administration, and the like). In the present application, "allows in vivo use" means that no toxicity is generated, or only a low level of toxicity of a pharmaceutically acceptable degree is generated, when administered to a living body (typically, when non-oral administration such as intraperitoneal administration, intramuscular administration, subcutaneous administration, and the like is performed in vivo). However, the non-lamellar liquid crystal-forming composition according to the present application is not limited to the purpose of in vivo use.

[0184] As the phospholipid used in the present application, one or more or more phospholipids selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, phosphatidic acid, and sphingomyelin, and salts thereof, or a phospholipid preparation or component containing the same, and the like can be exemplified, but are not limited thereto. As examples of phosphatidylcholine, dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), soybean phosphatidylcholine (SPC, also called soybean phospholipid), and egg yolk phosphatidylcholine (EPC, also called egg yolk lecithin), and the like can be exemplified, but are not limited thereto. As examples of phosphatidylethanolamine, dioleoylphosphatidylethanolamine (DOPE), and the like can be exemplified, but are not limited thereto. As examples of phosphatidylglycerol, dioleoylphosphatidylglycerol can be exemplified, and as examples of salts of phosphatidylglycerol, dioleoylphosphatidylglycerol sodium (DOPG-Na) can be exemplified, but are not limited thereto. The phospholipid used in the present application can be either a synthetic substance or a natural substance. In one embodiment, the phospholipid used in the present application can be phosphatidylcholine, for example, can be soybean phosphatidylcholine or egg yolk phosphatidylcholine. In another embodiment, the phospholipid used in the present application can be phosphatidylcholine, phosphatidylethanolamine, or phosphatidylglycerol, or a salt thereof. In one embodiment, the phospholipid used in the present application can be selected from the group consisting of soybean phosphatidylcholine (SPC) and egg yolk phosphatidylcholine (EPC), dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), and dioleoylphosphatidylethanolamine (DOPE). The non-lamellar liquid crystal-forming composition according to the present application can also contain one or more or more phospholipids.

[0185] In the aim of improving the biocompatibility of the amphiphilic compound and improving the safety of the non-lamellar liquid crystal-forming composition, the weight ratio of the isoprenoid-type amphiphilic compound represented by the general formula (I) and the phospholipid contained in the non-lamellar liquid crystal-forming composition according to the present application is preferably the amphiphilic compound: phospholipid = 90:10 to 10:90, for example, can be the amphiphilic compound: phospholipid = 80:20 to 20:80, 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, 45:55 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, but is not limited to the above weight ratios.

[0186] In one embodiment, in a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by General Formula (I) [in the formula, n = 2 and m = 2, R represents a hydrophilic group from which one hydroxyl group is removed from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and the like butanediol, and isopropylidene glycol)], and a phospholipid, the weight ratio is preferably the amphiphilic compound: phospholipid = 90:10 to 10:90, for example, it can be the amphiphilic compound: phospholipid = 80:20 to 20:80, 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, 45:55 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.

[0187] In one embodiment, in a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by General Formula (I) [in the formula, n = 1 and m = 2, R represents a hydrophilic group from which one hydroxyl group is removed from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and the like butanediol, and isopropylidene glycol)], and a phospholipid, the weight ratio is preferably the amphiphilic compound: phospholipid = 90:10 to 10:90, for example, it can be the amphiphilic compound: phospholipid = 80:20 to 20:80, 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, 45:55 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.

[0188] In one embodiment, in a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by General Formula (I) [in the formula, n = 2 and m = 1, R represents a hydrophilic group after removing 1 hydroxyl group from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and the like butanediol, and isopropylidene glycol)], and a phospholipid, the weight ratio is preferably the amphiphilic compound: phospholipid = 90:10 to 10:90, for example, it can be the amphiphilic compound: phospholipid = 80:20 to 20:80, 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, 45:55 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.

[0189] In one embodiment, in a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by General Formula (I) [in the formula, n = 2 and m = 1, R represents a hydrophilic group after removing 1 hydroxyl group from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, sorbitan, isosorbide, and a diol (for example, but not limited to, propylene glycol, ethylene glycol, diethylene glycol, 1,3-butanediol and the like butanediol, and isopropylidene glycol)], and a phospholipid, the weight ratio is preferably the amphiphilic compound: phospholipid = 90:10 to 10:90, for example, it can be the amphiphilic compound: phospholipid = 80:20 to 20:80, 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, 45:55 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.

[0190] In addition, regarding the weight ratio of the above amphiphilic compound and phospholipid, in the case where a plurality of isoprenoid-type amphiphilic compounds represented by the general formula (I) is used, the total amount (weight) of the plurality of isoprenoid-type amphiphilic compounds is calculated; in the case where a plurality of phospholipids is used, the total amount (weight) of the plurality of phospholipids is calculated. Note that in the present specification, the terms "weight" and "mass" are used interchangeably.

[0191] In alternative embodiments, the non-lamellar liquid crystal-forming composition according to the present application can be free of phospholipid, as long as it has sufficiently high biocompatibility (i.e., safety) depending on the use, dosage form, composition, etc. Thus, the present application also relates to a composition, particularly a non-lamellar liquid crystal-forming composition, containing an amphiphilic compound represented by the general formula (I). Such a non-lamellar liquid crystal-forming composition, for example, in the case where it contains a drug, can also be used as a sustained-release preparation.

[0192] In the present application, the "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, although it does not itself form a non-lamellar liquid crystal structure, has the ability to form a non-lamellar liquid crystal structure in the presence of water (i.e., by contact with an aqueous medium) (liquid crystal precursor composition).

[0193] When the non-lamellar liquid crystal-forming composition according to the present application is a liquid crystal precursor composition, it is free of an aqueous medium, or free of an aqueous medium in an amount sufficient to form a non-lamellar liquid crystal structure. When the non-lamellar liquid crystal-forming composition according to the present application is a non-lamellar liquid crystal composition, it contains an aqueous medium, preferably an aqueous medium in an amount sufficient to form a non-lamellar liquid crystal structure. The aqueous medium is not particularly limited and can be sterilized water, purified water, distilled water, ion-exchanged water, ultrapure water, water for injection, physiological saline, phosphate buffer, etc. The non-lamellar liquid crystal-forming composition according to the present application can also 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 application as a liquid crystal emulsion preferably further contains a surfactant. In addition, the non-lamellar liquid crystal emulsion composition is also referred to as a dispersion. The above non-lamellar liquid crystal emulsion composition according to the present application containing an amphiphilic compound and a phospholipid also exhibits high safety.

[0194] As an example of the surfactant used in the non-lamellar liquid crystal-forming composition according to the present application, P80 [polyoxyethylene sorbitan monooleate (20 E.O.)] can be given. As another example of the surfactant, nonionic surfactants represented by a block copolymer of a hydrophilic ethylene oxide and a hydrophobic propylene oxide (polyoxyethylene polyoxypropylene glycol), a polyoxyethylene alkyl ether, a polyoxyethylene alkyl ester, and a polyoxyethylene hardened castor oil can be given. As the nonionic surfactant, a nonionic surfactant having a molecular weight of 1000 or more (more preferably 5000 or more) is preferred. As the block copolymer of ethylene oxide and propylene oxide, polyoxyethylene (200) polyoxypropylene (70) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol, polyoxyethylene (160) polyoxypropylene (30) glycol, polyoxyethylene (120) polyoxypropylene (40) glycol, and the like can be given. These block copolymers of ethylene oxide and propylene oxide are commercially available under various names such as Pluronic (R) , Poloxamer (R) , Unilube (R) , propofol (R) , and the like. As a particularly preferred example of the nonionic surfactant, polyoxyethylene (200) polyoxypropylene (70) glycol, polyoxyethylene (196) polyoxypropylene (67) glycol (alias: Pluronic (R) F127, Unilube 70DP-950B, Poloxamer (R) 407), and the like can be given. Note that, in the present application, the amphiphilic compound represented by General Formulae (I) and (I’) used in the present application is not included in the range of the surfactant. The non-lamellar liquid crystal-forming composition according to the present application can also contain one or two or more of such surfactants.

[0195] The non-lamellar liquid crystal-forming composition according to the present application can also contain at least one of an oil and an organic solvent.

[0196] As an example of the oil used in the non-lamellar liquid crystal-forming composition according to the present application, but not limited to, vegetable oils such as sesame oil, soybean oil, corn oil, coconut oil, safflower oil, perilla oil, olive oil, castor oil, cottonseed oil, and the like; animal oils such as egg yolk oil, fish oil, lanolin, and the like; mineral oils such as medium-chain triglyceride (MCT), triglyceride, liquid paraffin, and the like; hydrocarbon oils such as squalene, squalane, and the like; ester oils such as isopropyl myristate (IPM) and the like; cholesterol; tocopherol; tocopherol acetate; glyceryl dioleate (GDO); gelled hydrocarbon; tetrahydrofarnesyl methyl acetate; hexahydrogeranylgeranyl methyl acetate, and the like can be given. The oil is preferably a pharmaceutically acceptable oil.

[0197] The total amount of the isoprenoid-type amphiphilic compound represented by the general formula (I), the phospholipid, and the oil contained in the non-lamellar liquid crystal-forming composition according to the present application can be, but is not limited to, 30% or more, typically 60 to 100%, preferably 65 to 95%, for example, 75 to 95%, 75 to 93%, or 80 to 95% or so, of the entire amount of the composition in the case of a liquid crystal precursor composition; and 0.01 to 40%, preferably 1 to 30% or so, for example, 20 to 30%, 20 to 23%, or 25 to 30% or so, of the entire amount of the composition in the case of a liquid crystal emulsion composition, depending on the purpose or the like.

[0198] Note that the related proportions (%) of the components in the non-lamellar liquid crystal-forming composition in the present specification mean weight %, which can be expressed in the unit of w / w %.

[0199] As examples of the organic solvent used in the non-lamellar liquid crystal-forming composition according to the present application, there can be mentioned, but are not limited to, alcohols such as ethanol, propylene glycol, and isopropyl alcohol; ethers such as diethyl ether and polyethylene glycol; dimethyl sulfoxide (DMSO); N-methylpyrrolidone (NMP); dimethylacetamide (DMA); and the like. The organic solvent is preferably a pharmaceutically acceptable organic solvent. In the non-lamellar liquid crystal-forming composition according to the present application, a protic organic solvent or an aprotic organic solvent can be used alone or a protic organic solvent and an aprotic organic solvent can be used in combination. As the protic organic solvent, there can be mentioned alcohols such as ethanol and propylene glycol; and polyethylene glycol; and the like. As the aprotic organic solvent, there can be mentioned ethers such as diethyl ether; dimethyl sulfoxide (DMSO); N-methylpyrrolidone (NMP); dimethylacetamide (DMA); and the like.

[0200] The non-lamellar liquid crystal-forming composition according to the present application can also contain a water-soluble polymer. As the water-soluble polymer, there can be mentioned, but not limited to, hydroxypropyl cellulose (HPC), hydroxyethyl cellulose, polyvinylpyrrolidone, polycarboxyvinyl, carrageenan, chitosan, chondroitin sulfate, xanthan gum, hyaluronic acid salt (sodium hyaluronate, etc.), alginic acid salt (sodium alginate, etc.), gelatin, dextran, etc. As the hydroxypropyl cellulose (HPC), there can be mentioned, for example, the following five grades of HPC marketed by 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 molecular weight of the hydroxypropyl cellulose can be 1000,000 or less, or 800,000 or less, for example, 10,000 to 700,000 or 10,000 to 80,000.

[0201] The non-lamellar liquid crystal-forming composition according to the present application can also contain an antioxidant. As the antioxidant, there can be mentioned, but not limited to, ascorbic acid, sodium sulfite, etc.

[0202] The non-lamellar liquid crystal-forming composition according to the present application can contain water, or can form a gel in the presence of water. By "form a gel" is meant to form a non-lamellar liquid crystal (liquid crystal gel). The non-lamellar liquid crystal can hold a substance such as a drug inside, and perform a sustained release. The present application also relates to a gel (gel-like composition) containing the non-lamellar liquid crystal-forming composition according to the present application.

[0203] The non-lamellar liquid crystal is a liquid crystal structure other than a lamellar liquid crystal, and specifically, for example, can be a cubic liquid crystal, a reverse hexagonal liquid crystal (HII), a reverse micellar cubic phase (Fd3m), or a sponge phase (L3). The non-lamellar liquid crystal can contain two or more non-lamellar liquid crystal phases.

[0204] The cubic liquid crystal can be a cubic liquid crystal belonging to crystal space group Ia3d (hereinafter referred to as "Ia3d cubic liquid crystal"), a cubic liquid crystal belonging to crystal space group Pn3m (hereinafter referred to as "Pn3m cubic liquid crystal"), or a cubic liquid crystal belonging to crystal space group Im3m (hereinafter referred to as "Im3m cubic liquid crystal").

[0205] The analysis of the liquid crystal structure can be performed by a conventional method, and for example, can be performed by small-angle X-ray scattering (SAXS) measurement using the following method.

[0206] When the sample is determined to be an emulsion, for example, the sample can be placed in an X-ray capillary made of sodium glass or quartz, and the like, and the capillary is sealed with an oxygen torch, and used for SAXS measurement. The SAXS measurement can be performed using a commercially available instrument, and for example, the measurement can be performed using a NANO Viewer nanometer-scale X-ray structure evaluation device (manufactured by Rigaku Corporation).

[0207] After the SAXS measurement is performed, it is confirmed whether or not the specific following scattering peak ratios (peak intervals) are exhibited in each liquid crystal structure, and thereby it can be confirmed whether or not the non-lamellar liquid crystal-forming composition according to the present application is formed, or the liquid crystal structure formed in the presence of water.

[0208] The ratio of the Pn3m cubic liquid crystal:

[0209] √2:√3:√4:√6:√8:√9:√10:,,,

[0210] The ratio of the Ia3d cubic liquid crystal:

[0211] √3:√4:√7:√8:√10:√11:,,,

[0212] The ratio of the Im3m cubic liquid crystal:

[0213] √2:√4:√6:√8:√10:√12:√14:,,,

[0214] The ratio of the Fd3m cubic liquid crystal:

[0215] √3:√8:√11:√12:√16:√19:√24:√27:,,,

[0216] The ratio specific to the inverse hexagonal liquid crystal:

[0217] 1:√3:2:,,,

[0218] In addition, the peak values can be calculated from the intensity distribution data of SAXS according to a method known to those skilled in the art, and the ratio of the reciprocals of these values can be calculated, and thereby the space group and the lattice constant can be easily determined.

[0219] On the other hand, in the SAXS measurement of the sponge phase (L3 phase), a broad scattering peak is observed.

[0220] The analysis of the scattering vector values of the peaks possessed by the sample to be measured, starting from the scattering vector value q1 [nm"1] of the peak located on the side of the smallest angle, makes it possible to not only determine the type of liquid crystal phase, but also to calculate the interplanar spacing and the lattice constant. Based on the results of the analysis, the composition of the non-lamellar liquid crystal-forming composition is changed, thereby changing the size of the liquid crystal phase and the unit cell, and thus the drug release properties (release rate) of the composition and preparation (e.g., a precursor preparation, an emulsion) containing the non-lamellar liquid crystal-forming composition and the drug can be controlled. For example, the size of the liquid crystal phase and the unit cell is changed according to the amphiphilic compound represented by the general formula (I), the phospholipid, and other components, and their proportions (weight ratio), thereby obtaining the corresponding release rates. When the amphiphilic compounds represented by the general formula (I) having similar R and the same carbon chain length (the same values of n and m), or similar carbon chain length (the same value of m only) and the same R are combined with the same phospholipid, and used in the non-lamellar liquid crystal-forming composition at the same degree of weight ratio relative to the phospholipid, they exhibit the same or similar liquid crystal phase, and exhibit the same drug release properties (release rate, etc.). In general, the release rate is faster in the order of reverse micellar cubic phase (Fd3m), reverse hexagonal liquid crystal (HII), and cubic liquid crystal (from low).

[0221] The non-lamellar liquid crystal-forming composition according to the present application can be used as a medical substrate for in vivo application. The non-lamellar liquid crystal-forming composition according to the present application can be used for preventing biological tissue adhesion because it has an effect of preventing biological tissue adhesion. Therefore, the non-lamellar liquid crystal-forming composition according to the present application can be used for preventing biological tissue adhesion. The non-lamellar liquid crystal-forming composition according to the present application can be used (applied) in vivo as a biological tissue adhesion preventing agent.

[0222] The non-lamellar liquid crystal-forming composition according to the present application can prevent biological tissue adhesion by being applied to biological tissue where adhesion is likely to occur. In the present application, the "anti-adhesion effect" refers to an effect of preventing the adhesion of a tissue having the potential for adhesion to other tissues or organs, thereby making it difficult to peel off, and completely or at a low level inhibiting the adhesion. Therefore, the non-lamellar liquid crystal-forming composition according to the present application can be used for preventing biological tissue adhesion.

[0223] The anti-adhesion effect of the non-lamellar liquid crystal-forming composition according to the present application can be achieved by the formation of a non-lamellar liquid crystal on the surface of the tissue of the application target by the amphiphilic compound contained in the non-lamellar liquid crystal-forming composition, thereby forming an envelope. The envelope formed can prevent the contact of the tissue with other tissues or organs, thereby reducing the adhesion.

[0224] The anti-adhesion effect of the non-lamellar liquid crystal-forming composition according to the present application can be confirmed, for example, by applying the non-lamellar liquid crystal-forming composition to the incision of the tissue of an open abdominal animal model, and observing the progress after closing the abdomen. Specifically, the abdomen of a rat is incised in the midline (e.g., about 30 mm) to perform an open abdominal surgery, and the left and right upper abdominal wall parietal peritoneum is incised about 20 mm. After complete hemostasis, the peritoneal incision is continuously sutured and closed (e.g., using 5-0 silk thread). Thereafter, to either one of the left and right peritoneal incisions, a sample of the non-lamellar liquid crystal-forming composition is applied to cover the incision suture. When the non-lamellar liquid crystal-forming composition is a liquid crystal precursor composition, in order to induce the formation of liquid crystals, an aqueous medium (sterile water or the like) can also be added to the application site in the form of a spray or the like. To the other peritoneal incision, nothing is applied. Then, the abdominal wall is closed in two layers. After a certain period of time from the surgery (e.g., 7 days), the abdomen is opened, and whether or not adhesion is observed at the incision suture is evaluated.

[0225] The application of the non-lamellar liquid crystal-forming composition is performed according to a method corresponding to the dosage form thereof. Preferably, the amount of the non-lamellar liquid crystal-forming composition applied in the evaluation follows the above-mentioned conversion amount of the amphiphilic compound, and is typically an amount of 5 to 50 mg.

[0226] The adhesion evaluation can be performed, for example, by giving a score related to the adhesion strength as follows.

[0227] • 0: no adhesion

[0228] • 1: slightly pulling adhesion (without tissue damage)

[0229] • 2: strongly pulling adhesion (without tissue damage)

[0230] • 3: strongly pulling adhesion causing tissue damage

[0231] In the incision to which the sample of the non-lamellar liquid crystal-forming composition is applied, if the score is lower than that of the non-application incision of the same animal, it can be judged that the anti-adhesion effect is confirmed.

[0232] In addition, for example, the adhesion range ratio of each incision is calculated as the proportion (%) of the adhesion length with respect to the incision suture of about 20 mm, and based on this, the proportion of the adhesion range of the incision to which the sample of the non-lamellar liquid crystal-forming composition is applied with respect to the non-application incision (adhesion range ratio of the sample application side / adhesion range ratio of the non-application side x 100) can be judged as follows: 40% or less is A; 41% to 60% is B+; 61% to 80% is B-; and 81% or more is C. In this case, as the anti-adhesion effect, the judgment of A or B+ is preferred.

[0233] In the present application, "prevention of adhesion" can be judged by the following method: by applying (treating) the non-lamellar liquid crystal-forming composition, the frequency and / or degree of adhesion at the application site is reduced as compared with a control group which is not treated.

[0234] Note that it has been confirmed in Patent Literature 3 and the like that many of the amphiphilic compounds represented by General Formula (I) can exert an adhesion-preventing effect.

[0235] The present application also provides a method for preventing adhesion of biological tissue, comprising applying the non-lamellar liquid crystal-forming composition according to the present application to the biological tissue. More specifically, the present application also provides a method for preventing adhesion of a diseased tissue, comprising applying an effective amount of the non-lamellar liquid crystal-forming composition according to the present application to a diseased site of a patient, specifically a site where adhesion is likely to occur, specifically a site where tissue repair is expected to occur (e.g., an inflammation site or a damage site in the body). As specific examples of the above-mentioned site where adhesion is likely to occur, there can be mentioned an inflammation site in the body, whether exogenous or endogenous, a wound site such as an incision site during surgery, a site where the surface of tissue has been damaged by human handling such as touching during surgery, and the like. In the present application, the "damage site" refers to a portion of tissue or organ which has been damaged by surgery, trauma, disease, or the like. As examples of the tissue or organ to which the non-lamellar liquid crystal-forming composition is applied, there can be mentioned, but not limited to, peritoneum, small intestine, large intestine, rectum, stomach, duodenum, cecum, liver, uterus, oviduct, lymphatic vessel, heart, pericardium, lung, brain, ovary, tendon, and the like. As a typical example, the non-lamellar liquid crystal-forming composition according to the present application is applied to an incision site, the periphery of the incision site, or the entire organ having the incision site at the time of surgery. The non-lamellar liquid crystal-forming composition according to the present application can also be applied to a site in the body which comes into contact with a wound site, an inflammation site, or the like.

[0236] The non-lamellar liquid crystal-forming composition can be applied to the affected area, such as the site of injury (e.g., a wound site) and the site of inflammation, using a method corresponding to the dosage form. For example, the non-lamellar liquid crystal-forming composition can be sprayed on the affected area, such as the site of injury (e.g., a wound site) and the site of inflammation, using a gas-jet aerosol container. Alternatively, if the container is a pump spray, the non-lamellar liquid crystal-forming composition can be sprayed on the affected area, such as the site of injury (e.g., a wound site) and the site of inflammation, using a non-gas-jet spray container, such as a conventional hand-held pump spray. In the case of endoscopic surgery or laparoscopic surgery, the non-lamellar liquid crystal-forming composition can be sprayed on the affected area, such as the site of injury (e.g., a wound site), using a nozzle or the like used in endoscopic surgery or laparoscopic surgery. In the present application, "spraying" means that the target substance is ejected (sprayed and / or sprayed) in the form of droplets, mist, fine particles, or foam, or the like, by applying pressure. If the non-lamellar liquid crystal-forming composition is a coating agent, the composition can be applied to the affected area, such as the site of injury (e.g., a wound site) and the site of inflammation, in an appropriate amount. If the non-lamellar liquid crystal-forming composition is an injection, the composition can be injected into the affected area, such as the site of injury (e.g., a wound site) and the site of inflammation.

[0237] The non-lamellar liquid crystal-forming composition according to the present application is preferably applied to the affected area, such as the site of injury (e.g., a wound site) and the site of inflammation, in an amount sufficient to cover the affected area. Preferably, in one embodiment, if the application subject is a human, the non-lamellar liquid crystal-forming composition according to the present application is applied in an amount of 10 mg to 100 g or 50 mg to 50 g (more preferably 0.1 g to 10 g).

[0238] When the non-lamellar liquid crystal-forming composition according to the present application contains a sufficient amount of an aqueous medium (e.g., is a liquid crystal emulsion), the non-lamellar liquid crystal-forming composition can form a non-lamellar liquid crystal on the surface of the tissue to which it is applied. When the non-lamellar liquid crystal-forming composition according to the present application does not contain a sufficient amount of an aqueous medium (e.g., is a liquid crystal precursor composition), although a non-lamellar liquid crystal can be formed by moisture in the body, in order to promote the formation of the envelope, it is preferable to apply an aqueous medium to the affected site, such as the site of an injury (e.g., a wound site) and the site of inflammation, in addition to the non-lamellar liquid crystal-forming composition. The aqueous medium can be, for example, sterilized water, purified water, distilled water, ion-exchanged water, ultrapure water, water for injection, and the like, and can also be a physiologically acceptable aqueous solution. As the physiologically acceptable aqueous solution, for example, there can be mentioned physiological saline; aqueous electrolyte solutions such as aqueous sodium chloride solution, aqueous calcium chloride solution, aqueous magnesium chloride solution, aqueous sodium sulfate solution, aqueous potassium sulfate solution, aqueous sodium carbonate solution, and aqueous sodium acetate solution; buffer solutions such as phosphate buffer and Tris hydrochloride buffer; aqueous solutions containing sugar molecules such as glucose, sucrose, maltose, and hyaluronic acid; aqueous solutions containing water-soluble polymers such as polyethylene glycol and polyvinyl alcohol; and the like. As a preferable example of the physiologically acceptable aqueous solution, there can be mentioned an aqueous hyaluronic acid solution containing hyaluronic acid or a salt thereof (sodium hyaluronate, etc.).

[0239] After the application of the non-lamellar liquid crystal-forming composition as a liquid crystal precursor composition, it is preferable to apply an aqueous medium to the non-lamellar liquid crystal-forming composition, but it is not limited thereto. The aqueous medium can be applied by the same application method as the non-lamellar liquid crystal-forming composition, such as spraying, coating, or injection, and the like. After the application of the aqueous medium to the tissue or organ in this way, it is preferable to stand for a prescribed period of time (e.g., 1 to 30 minutes, preferably 5 to 10 minutes, but it is not limited thereto) in order to promote the formation of the envelope.

[0240] The subject (patient) to which the adhesion-preventing method using the non-lamellar liquid crystal-forming composition according to the present application is applied is typically a mammal such as a human, a domestic animal, a pet, and an experimental animal. It is particularly preferable to be a subject in which a tissue (organ) has been damaged or is expected to be damaged due to a surgical operation, an external injury, a disease, and the like. The surgical operation includes not only an open surgery but also an endoscopic surgery and a laparoscopic surgery, and the like.

[0241] The non-lamellar liquid crystal-forming composition according to the present application has significantly reduced or eliminated toxicity, and thus the adhesion-preventing method according to the present application has high safety for the patient to be treated.

[0242] The non-lamellar liquid crystal forming composition according to the present application can contain a drug in addition to the amphiphilic compound represented by the general formula (I) and the above-mentioned components such as phospholipid, as necessary. In the present application, the drug refers to any substance (effective ingredient) to be administered to a living body, for the purpose of maintaining and releasing (controlled release) in a non-lamellar liquid crystal structure by containing the drug in the non-lamellar liquid crystal forming composition. However, the drug is not the amphiphilic compound represented by the general formula (I) itself. The drug can be either an organic compound or an inorganic compound. The drug can be either a water-soluble drug or a fat-soluble (oleophilic, water-insoluble or water-poor soluble) drug. The drug can be a physiologically active substance. The drug can be, for example, a protein, a peptide, an amino acid, a nucleic acid, etc., but is not limited thereto. The drug can be, for example, a gonadotropin-releasing hormone (GnRH) agonist, but is not limited thereto. The gonadotropin-releasing hormone (GnRH) agonist can be, for example, leuprolide or a salt thereof, but is not limited thereto. The salt of leuprolide can be any pharmaceutically acceptable salt, and a carboxylic acid salt represented by leuprolide acetate (i.e., leuprolide acetate) can be mentioned, but is not limited thereto. In addition, leuprolide acetate is sometimes called by other names such as leuprolide acetate. Such a non-lamellar liquid crystal forming composition can be used for the sustained release of a drug.

[0243] The present application also provides a pharmaceutical preparation containing the non-lamellar liquid crystal forming composition according to the present application. The pharmaceutical preparation according to the present application preferably contains a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by the general formula (I) and a phospholipid, the phospholipid improving the biocompatibility of the non-lamellar liquid crystal forming composition. In one embodiment, the pharmaceutical preparation according to the present application contains a non-lamellar liquid crystal forming composition containing an amphiphilic compound represented by the general formula (I), a phospholipid and a drug. The "pharmaceutical preparation" in the present application can be a pharmaceutical composition. The pharmaceutical preparation or pharmaceutical composition in the present application can contain a pharmaceutically acceptable additive (e.g., a carrier, an excipient, a lubricant, a disintegrant, a humectant, a buffer, a flavoring agent, a preservative, a coloring agent, an odorant, a propellant, etc.) and other substances, as long as the non-lamellar liquid crystal forming ability is maintained.

[0244] The pharmaceutical preparation according to the present application can be a preparation in any dosage form, and can be, for example, a spray, an aerosol, an injection or a long-acting agent, etc.

[0245] The pharmaceutical preparation according to the present application can be used for preventing the adhesion of biological tissues. The pharmaceutical preparation according to the present application can be a sustained release preparation such as a long-acting agent, etc., further containing the above-mentioned drug.

[0246] The present application also provides a method for sustained-release delivery of a drug in a living body (in vivo) or in a living cell or a living tissue, which comprises: applying a drug preparation containing a drug according to the present application, or a non-lamellar liquid crystal-forming composition containing a drug according to the present application, to a living body such as a subject (patient) or the like, for example, in vivo (particularly, in a biological tissue in vivo) or on the surface of the body, or to a living cell or a living tissue. When a non-lamellar liquid crystal-forming composition or a drug preparation containing the same is used as a liquid crystal precursor composition, after the application, an aqueous medium can be applied to the non-lamellar liquid crystal-forming composition or the drug preparation, but is not limited thereto, and a liquid crystal can also be formed using moisture in the living body. Among them, the application to a living body (in vivo or on the surface of the body) or a living cell or a living tissue is preferably performed by non-oral administration (for example, intravenous, intra-arterial, intraperitoneal, intramuscular, subcutaneous, intradermal, or the like). By the method of the present application, a drug can be sustained-release delivered in vivo or in a living cell or a living tissue with high safety.

[0247] [Examples]

[0248] The present application is more specifically described below using examples. However, the technical scope of the present application is not limited to these examples.

[0249] [Example 1] Synthesis of amphiphilic compound (1)

[0250] (1) Synthesis of mono O-(5,9,13-trimethyltetradec-4-enoyl) sorbitol

[0251] [Chemical Formula 9]

[0252]

[0253] After 14.1 g (50.0 mmol) of 5,9,13-trimethyltetradec-4-enoic acid methyl ester and 10.9 g (60.0 mmol) of 90% by weight aqueous sorbitol (Sorbitol M-90, Sanoh Chemical Industry Co., Ltd., containing 90% of solid content and 10% of moisture, content in solid content: sorbitan 79-84%, isosorbide 15-18%) were added to a reaction vessel at room temperature, stirring was performed at 120°C, 8 kPa for 1 hour. After the pressure was released with nitrogen, 0.27 g (5.0 mmol) of sodium methoxide and 0.01 g of sodium hypophosphite monohydrate were added, and stirring was performed at 160°C, 8 kPa for 1 hour. After the pressure was released with nitrogen, 0.27 g (5.0 mmol) of sodium methoxide was added, and stirring was performed at 160°C, 8 kPa for 1 hour. After cooling to 60°C, 50 mL of ethyl acetate and 50 mL of 0.5 M hydrochloric acid were added while stirring. To the resulting reaction solution, 150 mL of ethyl acetate was added, and extraction was performed. The extracted solution was washed with saturated sodium bicarbonate water, saturated brine, dried with magnesium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to obtain 8.96 g (yield 43%) of a component containing the title compound as a light brown transparent liquid. The resulting component contained mono-O-(5,9,13-trimethyltetradec-4-enoyl)sorbitan and mono-O-(5,9,13-trimethyltetradec-4-enoyl)isosorbide in a ratio of about 8:2 (weight ratio) (determined by GC-MS in ion mode EI+by area value of TIC, and calculated from the area value). The resulting component also contained a small amount of a diester derivative from sorbitan (presumed by GC-MS determination and TLC analysis). Regarding the resulting component, 1 The measurement results of H-NMR are shown below.

[0254] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 9H), 0.9-1.7 (m, 15H), 1.8-2.0 (m, 2H), 2.2-2.5 (m, 4H), 3.5-4.9 (m, 6.1H), 5.04 (m, 1H), 5.0-5.2 (m, 0.6H)

[0255] Mono-O-(5,9,13-trimethyltetradec-4-enoyl)sorbitan is also referred to as C17 sorbitan ester.

[0256] The resulting component was used as a mono-O-(5,9,13-trimethyltetradec-4-enoyl)sorbitan component (C17 sorbitan ester component) in the examples described later.

[0257] (2) Synthesis of mono O-(5, 9, 13-trimethyltetradecanoyl) sorbitan

[0258] [Chem. 10]

[0259]

[0260] To a solution of 4.14 g (10.0 mmol) of mono O-(5, 9, 13-trimethyltetradec-4-enoyl) sorbitan in ethyl acetate (12 mL) was added 0.48 g of 5% palladium on carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred for 2 days under an ordinary pressure hydrogen gas atmosphere at room temperature. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (mobile phase: ethyl acetate) to thereby obtain 4.02 g of a component containing the title compound as a colorless transparent liquid (yield 97%). The obtained component contained mono O-(5, 9, 13-trimethyltetradecanoyl) sorbitan and mono O-(5, 9, 13-trimethyltetradecanoyl) isosorbitol in a ratio of about 8:2 (weight ratio) (determined by GC-MS in the ion mode EI+by the area value of TIC, and calculated from the area value). The obtained component also contained a small amount of a diester derivative from sorbitan (presumed by GC-MS measurement and TLC analysis). As for the obtained component, 1 The measurement results of H-NMR are shown below.

[0261] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 12H), 0.9-1.7 (m, 19H), 2.2-2.4 (m, 2H), 3.5-4.9 (m, 6.4H), 5.0-5.2 (m, 0.6H)

[0262] Mono O-(5, 9, 13-trimethyltetradecanoyl) sorbitan is also called saturated C17 sorbitan ester.

[0263] (3) Synthesis of mono O-(5, 9, 13, 17-tetramethyloctadec-4-enoyl) sorbitan

[0264] [Chem. 11]

[0265]

[0266] After 70.5 g (200 mmol) of 5, 9, 13, 17-tetramethyloctadeca-4- enoic acid methyl ester and 54.7 g (300 mmol) of 90 wt% aqueous sorbitol (Sorbitol M-90, Sanoh Chemical Industry Co., Ltd., containing 90% solid content and 10% moisture, content in solid content: sorbitan 79-84%, isosorbide 15-18%) were added to a reaction vessel at room temperature, stirring was performed at 120°C, 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 2.2 g (40 mmol) of sodium methoxide and 0.04 g of sodium hypophosphite monohydrate were added, and stirring was performed at 160°C, 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 1.1 g (20 mmol) of sodium methoxide was added, and stirring was performed at 160°C, 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 1.1 g (20 mmol) of sodium methoxide was further added, and stirring was performed at 160°C, 8 kPa for 1.5 hours. After cooling to 60°C, 200 mL of ethyl acetate and 200 mL of 0.5 M hydrochloric acid were added while stirring. To the resulting reaction solution, 600 mL of ethyl acetate was added, and extraction was performed. The extracted solution was washed with saturated sodium bicarbonate water, saturated brine, dried with magnesium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to obtain 36.1 g (yield 37%) of a component containing the title compound as a light brown transparent liquid. The resulting component contained mono-O- (5, 9, 13, 17-tetramethyloctadeca-4-enoyl) sorbitan and mono-O- (5, 9, 13, 17-tetramethyloctadeca-4-enoyl) isosorbide in a ratio of about 8:2 (weight ratio) (determined by GC-MS in ion mode EI+by area value of TIC, and calculated from the area value). The resulting component also contained a small amount of a diester derivative from sorbitan (presumed by GC-MS determination and TLC analysis). Regarding the resulting component, 1 The measurement results of H-NMR are shown below.

[0267] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 12H), 0.9-1.8 (m, 22H), 1.85-2.0 (m, 2H), 2.0-2.5 (m, 4H), 3.5-4.9 (m, 6.4H), 5.04 (m, 1H), 5.0-5.2 (m, 0.6H)

[0268] Mono-O- (5, 9, 13, 17-tetramethyloctadeca-4-enoyl) sorbitan is also called C22 sorbitan ester.

[0269] The obtained component was used as a mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) sorbitan component (C22 sorbitan ester component) in the Examples described later.

[0270] In addition, 13.00 g of the C22 sorbitan ester component was subjected to silica gel column purification (mobile phase: ethyl acetate / hexane mixture) to remove low-polarity components such as diester derivatives from sorbitan from the C22 sorbitan ester component and mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) isosorbide, whereby 6.78 g of a purified component was obtained as a slightly yellow transparent liquid. The purity thereof was 99% or more of mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) sorbitan (area value of TIC determined by GC-MS in ion mode EI+, calculated from the area value). In addition, it was almost free of low-polarity components such as diester derivatives from sorbitan (presumed by GC-MS determination and TLC analysis). Regarding the obtained compound, 1 The measurement results of H-NMR are shown below.

[0271] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.8-0.9 (m, 12H), 0.9-1.7 (m, 22H), 1.9-2.0 (m, 2H), 2.2-2.5 (m, 4H), 2.93 (br s, OH), 3.6-4.5 (m, 7.5H), 5.06 (br d, J=5.0 Hz, IH), 4.9-5.2 (m, 0.5H)

[0272] The purified component obtained as above was called high-purity mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) sorbitan, or high-purity C22 sorbitan ester.

[0273] (4) Synthesis of mono-O-(5,9,13,17-tetramethyloctadecanoyl) sorbitan

[0274] [Chemical Formula 12]

[0275]

[0276] To a solution of 5.82 g (12.0 mmol) of the C22 sorbitan ester component in ethyl acetate (17.4 mL) was added 0.70 g of 5% palladium on carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred for 2 days at room temperature under an ordinary pressure hydrogen gas atmosphere. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (mobile phase: ethyl acetate) to thereby obtain a component containing the title compound as a colorless transparent liquid 5.69 g (yield 97%). The obtained component contained mono-O- (5, 9, 13, 17-tetramethyloctadecanoyl) sorbitan and mono-O- (5, 9, 13, 17-tetramethyloctadecanoyl) isosorbitol in a ratio of about 8:2 (weight ratio) (determined by GC-MS in the ion mode EI+by the area value of TIC, and calculated from the area value). The obtained component also contained a small amount of a diester derivative from sorbitan (presumed by GC-MS measurement and TLC analysis). As for the obtained component, 1 The measurement results of H-NMR are shown below.

[0277] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 15H), 0.9-1.7 (m, 26H), 2.2-2.4 (m, 2H), 3.5-4.9 (m, 6.4H), 5.0-5.2 (m, 0.6H)

[0278] Mono-O- (5, 9, 13, 17-tetramethyloctadecanoyl) sorbitan is also called saturated C22 sorbitan ester.

[0279] The obtained component was used as a mono-O- (5, 9, 13, 17-tetramethyloctadecanoyl) sorbitan component (saturated C22 sorbitan ester component) in the examples described later.

[0280] (5) Synthesis of mono-O- (5, 9, 13, 17-tetramethyloctadec-4-enoyl) isosorbitol

[0281] [Chemical Formula 13]

[0282]

[0283] At room temperature, 35.3 g (100 mmol) of 5,9,13,17-tetramethyloctadeca-4- enoic acid methyl ester and 29.2 g (200 mmol) of isosorbide (Sanwa Chemical Co., Ltd., purity 98.0% or more, moisture 1.0% or less) were added to a reaction vessel. After warming to 120°C while stirring at 8 kPa, the pressure was released with nitrogen, and 1.1 g (20 mmol) of sodium methoxide and 0.02 g of sodium hypophosphite monohydrate were added. After warming to 180°C under a nitrogen atmosphere, the pressure was released to 8 kPa and stirring was performed for 1 hour. The pressure was released with nitrogen, and 0.54 g (10 mmol) of sodium methoxide was added, and stirring was performed again at 180°C, 8 kPa for 1 hour. The pressure was released with nitrogen, and 0.54 g (10 mmol) of sodium methoxide was added again, and stirring was performed again at 180°C, 8 kPa for 1 hour. After cooling to 60°C, 100 mL of ethyl acetate and 100 mL of 0.5 M hydrochloric acid were added while stirring. To the resulting reaction solution, 300 mL of ethyl acetate was added, and extraction was performed. The extracted solution was washed with saturated sodium bicarbonate water, saturated brine, dried with magnesium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to obtain 17.9 g (yield 38%) of the title compound as a light brown transparent liquid.

[0284] This compound was also purified by silica gel column chromatography (ethyl acetate / hexane mixture) to separate 2-O-(5,9,13,17-tetramethyloctadeca-4-enoyl) isosorbide and 5-O-(5,9,13,17-tetramethyloctadeca-4-enoyl) isosorbide in a ratio of 60:40 (weight ratio). Regarding the two compounds obtained, 1 The measurement results of H-NMR are shown below.

[0285] <2-O-(5,9,13,17-tetramethyloctadeca-4-enoyl) isosorbide>

[0286] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 12H), 0.9-1.7 (m, 22H), 1.85-2.0 (m, 2H), 2.2-2.4 (m, 4H), 2.56 (d, J=7.2 Hz, OH), 3.55 (dd, J=6.0, 9.5 Hz, 1H), 3.87 (dd, J=6.0, 9.5 Hz, 1H), 3.99 (m, 2H), 4.29 (m, 1H), 4.44 (d, J=4.3 Hz, 1H), 4.60 (t, J=4.9 Hz, 1H), 5.04 (m, 1H), 5.21 (s, 1H)

[0287] <5-O-(5,9,13,17-tetramethyloctadec-4-enoyl) isosorbide>

[0288] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 12H), 0.9-1.7 (m, 22H), 1.83 (br s, OH), 1.85-2.0 (m, 2H), 2.2-2.4 (m, 4H), 2.2-2.4 (m, 4H), 3.74 (ddd, J = 2.1.5.2, 9.8 Hz, IH), 3.82-3.93 (m, 3H), 4.31 (s, IH), 4.38 (d, J = 4.6 Hz, IH), 4.83 (t, J = 5.0 Hz, IH), 5.07 (m, IH), 5.13 (m, IH)

[0289] Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) isosorbide is also called C22 isosorbide ester.

[0290] (6) Synthesis of Mono-O-(5,9,13,17-tetramethyloctadecanoyl) isosorbide

[0291] [Chemical 14]

[0292]

[0293] To a solution of 4.66 g (10.0 mmol) of Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) isosorbide in ethyl acetate (14 mL) was added 0.56 g of 5% palladium on carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred for 2 days at room temperature under a normal pressure hydrogen gas atmosphere. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (mobile phase: ethyl acetate) to thereby obtain the title compound 4.65 g (yield 99%) as a colorless transparent liquid.

[0294] Mono-O-(5,9,13,17-tetramethyloctadecanoyl) isosorbide is also called saturated C22 isosorbide ester.

[0295] (7) Synthesis of Mono-O-(5,9,13-trimethyltetradec-4-enoyl) glycerol

[0296] [Chemical 15]

[0297]

[0298] To a solution of 0.65 g (7.1 mmol) of glycerol and 0.59 g (4.3 mmol) of potassium carbonate in dry N,N-dimethylformamide (3.5 mL) was slowly added 1.0 g (3.5 mmol) of 5,9,13-trimethyltetradec-4-enoic acid methyl ester (tetrahydrofarnesyl acetic acid methyl ester) dropwise at 80°C. After stirring at 100°C for 18 hours, 1 M hydrochloric acid was added to the reaction solution, and extraction was performed with ether. The extract was washed successively with saturated aqueous sodium bicarbonate, saturated aqueous sodium chloride, and dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to thereby obtain the title compound as a colorless transparent liquid. Regarding the obtained compound, 1 The results of the H-NMR measurement and the viscosity measurement are shown below.

[0299] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.80-0.90 (m, 9H), 1.00-1.70 (m, 15H), 1.97 (td, J = 7.8, 17.0 Hz, 2H), 2.13 (t, J = 6.1 Hz, IH, OH), 2.25-2.45 (m, 4H), 2.55 (d, J = 5.2 Hz, IH, OH), 3.50-4.00 (m, 3H), 4.10-4.25 (m, 2H), 5.08 (t, J = 6.7 Hz, IH)

[0300] Viscosity: 0.48 Pa-s (shear rate 92 1 / s)

[0301] Mono 0-(5,9,13-trimethyltetradec-4-enoyl) glycerol is also called C17 glyceride.

[0302] (8) Synthesis of mono 0-(5,9,13,17-tetramethyloctadec-4-enoyl) glycerol

[0303] [Chemical 16]

[0304]

[0305] To a solution of 23.5 g (255 mmol) of glycerol and 0.55 g (4.0 mmol) of potassium carbonate in dry N,N-dimethylformamide (48 mL) was added 28.2 g (80.0 mmol) of 5,9,13,17-tetramethyloctadeca-4- enoic acid methyl ester slowly dropwise at 80°C under reduced pressure of 60 to 70 mmHg and a stream of nitrogen, and stirred for 3 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 200 mL), washed with water, saturated sodium hydrogen carbonate aqueous solution, saturated brine (2 times), and 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 30:70), to thereby obtain the title compound 13.3 g (yield 40%) as a slightly yellow transparent liquid. The obtained compound was identified by1H-NMR. 1 The measurement results of1H-NMR are shown below.

[0306] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 1.00-1.70 (m, 22H), 1.85-2.15 (m, 2H), 2.15-2.55 (m, 4H), 3.53-3.78 (m, 3H), 3.80-4.00 (m, IH), 4.10-4.25 (m, 2H), 5.08 (dd, J = 6.9 Hz, J = 6.9 Hz, IH)

[0307] Mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol is also called C22 glyceride.

[0308] (9) Synthesis of Mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol

[0309] [Chemical 17]

[0310]

[0311] To a solution of 20.6 g (50.0 mmol) of mono-O-(5,9,13,17-tetramethyloctadeca-4- enoyl)glycerol in ethyl acetate (62 mL) was added 2.5 g of 5% palladium carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred for 42 hours at room temperature under an ordinary pressure hydrogen gas atmosphere. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium carbon was filtered off. The filtrate was purified by silica gel column chromatography (ethyl acetate), to thereby obtain the title compound 20.2 g (yield 98%) as a colorless transparent liquid. The obtained compound was identified by1H-NMR. 1 The measurement results of1H-NMR are shown below.

[0312] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.7-0.9 (m, 15H), 0.95-1.75 (m, 26H), 2.13 (t, J = 6.0 Hz, OH), 2.34 (t, J = 7.7 Hz, 2H), 2.56 (d, J = 5.1 Hz, OH), 3.55-3.75 (m, 2H), 3.94 (m, IH), 4.15 (dd, J = 6.0, 11.7 Hz, IH), 4.20 (dd, J = 4.7, 11.7 Hz, IH)

[0313] Mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol is also called saturated C22 glyceride.

[0314] (10) Synthesis of Mono-O-(5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoyl)glycerol

[0315] [Chemical 18]

[0316]

[0317] To a solution of 58.1 g (200 mmol) of 3,7,11,15-tetramethylhexadeca-1,6,10,14-tetraen-3-ol (geranyl linalool) and 19 mL (0.15 mol) of trimethyl orthoacetate, 53 mL (0.42 mol) of trimethyl orthoacetate and 5.0 mL (40 mmol) of n-hexanoic acid were added dropwise at 135°C for 8 hours under a nitrogen atmosphere. After stirring at the same temperature for 6 hours, a solution of 5.3 mL (42 mmol) of trimethyl orthoacetate and 0.5 mL (4 mmol) of n-hexanoic acid was added dropwise and stirred at the same temperature for 2 hours. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (3:1, 300 mL), washed with saturated sodium bicarbonate water (2 times), saturated brine, and dried over magnesium sulfate. After filtration, concentration was performed, whereby 67.24 g of methyl 5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoate (methyl geranyl geranate) was obtained as a crude product liquid. This crude product was directly used in the following reaction.

[0318] To a solution of 7.4 g (80 mmol) of glycerol and 5.5 g (40 mmol) of potassium carbonate in dry N,N-dimethylformamide (16 mL) was added 13.9 g (40.0 mmol) of 5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoic acid methyl ester (methyl geranylgeranoate) dropwise at 85°C under reduced pressure of 200 to 250 mmHg, and stirred for 6 hours at the same temperature. During this time, the methanol produced by the reaction was distilled off. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 200 mL), washed with water, saturated sodium bicarbonate aqueous solution, saturated brine (2 times), and dried over magnesium sulfate. The residue obtained by concentration after filtration was purified by silica gel column chromatography (mobile phase: hexane / ethyl acetate = 100:0 to 0:100), whereby 5.44 g (yield 33%) of the title compound was obtained as a transparent liquid. The obtained compound, 1 The results of H-NMR measurement and viscosity measurement are shown below.

[0319] 1 H-NMR spectrum (270 MHz, CDC13, TMS) δ: 1.55-1.72 (m, 15H), 1.9-2.2 (m, 13H), 2.27-2.45 (m, 4H), 2.53 (br s, IH, OH), 3.59 (dd, J = 5.4, 11.4 Hz, IH), 3.68 (dd, J = 3, 11.4 Hz, IH), 3.92 (m, IH), 4.15 (dd, J = 6.0, 11.6 Hz, IH), 4.21 (dd, J = 4.8, 11.6 Hz, IH), 5.05-5.15 (m, 4H)

[0320] Viscosity: 0.37 Pa-s (shear rate 92 1 / s)

[0321] Mono 0-(5,9,13,17-tetramethyloctadeca-4,8,12,16-tetraenoyl) glycerol is also called geranylgeranyl acetin.

[0322] (11) Synthesis of mono 0-(5,9,13,17-tetramethyloctadecanoyl) erythritol

[0323] [Chemical Formula 19]

[0324]

[0325] To 10 g of 5, 9, 13, 17-tetramethyloctadecanoic acid, 20 ml of dichloromethane was added 1 drop of pyridine under nitrogen atmosphere, and 5.2 g of thionyl chloride was added dropwise at room temperature. After the completion of the dropwise addition, it was refluxed for 1 hour, and concentrated under reduced pressure to obtain 10.5 g of 5, 9, 13, 17-tetramethyloctadecanoic acid chloride.

[0326] To 2.56 g of erythritol, 2.21 g of pyridine, 70 ml of dry DMF was added and dissolved by heating. After cooling to room temperature, a solution of 5 g of 5, 9, 13, 17-tetramethyloctadecanoic acid chloride obtained above dissolved in 10 ml of dichloromethane was added dropwise, and stirred at room temperature for 1 hour after the dropwise addition. To the resulting reaction solution, 100 ml of dichloromethane was added, washed with saturated brine 3 times, and dried over anhydrous sodium sulfate. After filtration and concentration under reduced pressure, it was purified by silica gel column chromatography to obtain 2.83 g of the title compound as a transparent semisolid. HPLC analysis showed that the title compound contained 91.6% of 1-O- (5, 9, 13, 17-tetramethyloctadecanoyl) erythritol and 8.4% of 2-O- (5, 9, 13, 17-tetramethyloctadecanoyl) erythritol. In addition, with respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0327] 1 H-NMR spectrum (270 MHz, CDC13, TMS) δ: 0.8-0.9 (m, 15H), 1.0-1.7 (m, 26H), 2.11 (br. s, IH), 2.33 (t, J = 7.9 Hz, 2H), 2.66 (br. s, IH), 2.75 (br. s, IH), 3.6-3.9 (m, 4H), 4.29-4.36 (m, 2H)

[0328] Mono-O- (5, 9, 13, 17-tetramethyloctadecanoyl) erythritol is also called saturated C22 erythritol ester.

[0329] (12) Synthesis of mono-O- (5, 9, 13, 17-tetramethyloctadec-4-enoyl) pentaerythritol

[0330] [Chemical Formula 20]

[0331]

[0332] To a solution of 157 g (1.15 mol) of pentaerythritol and 1.58 g (1.15 mmol) of potassium carbonate in dry N,N-dimethylformamide (700 mL) was slowly added 250 g (0.71 mol) of 5,9,13,17-tetramethyloctadeca-4-enoic acid methyl ester at 78-83°C under reduced pressure of 60-70 mmHg and nitrogen flow. After stirring at the same temperature for 10 hours, formic acid was added at 75°C to adjust the pH to 4. The resulting solution was concentrated under reduced pressure, diluted with methyl tert-butyl ether (1.5 L), and the resulting insolubles were filtered off. The resulting filtrate was washed with 10% aqueous sodium bicarbonate twice, treated with activated carbon (8 g), and decolorized. The resulting filtrate was concentrated, and the residue was purified by silica gel column chromatography (mobile phase: hexane / ethyl acetate mixture) to obtain the title compound.

[0333] For the resulting compound, H-NMR measurement, infrared absorption (IR) spectrum measurement by infrared spectrophotometry, and viscosity measurement were performed. The measurement results are shown below. 1 H-NMR measurement, infrared absorption (IR) spectrum measurement by infrared spectrophotometry, and viscosity measurement were performed. The measurement results are shown below.

[0334] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 1.00-1.70 (m, 22H), 1.90-2.05 (m, 2H), 2.25-2.45 (m, 4H), 3.64 (s, 6H), 4.24 (s, 2H), 5.07 (br s, IH)

[0335] IR spectrum (NaCl film method): 3387, 2926, 2866, 1739, 1461, 1378, 1267, 1139, 1051

[0336] Viscosity: 1.7 Pa-s

[0337] Mono O-(5,9,13,17-tetramethyloctadecanoyl) pentaerythritol is also referred to as C22 pentaerythritol ester.

[0338] (13) Synthesis of mono O-(5,9,13,17-tetramethyloctadecanoyl) pentaerythritol

[0339] [Chemical Formula 21]

[0340]

[0341] A mixture of 3.81 g of pentaerythritol, 2.21 g of pyridine, and 120 mL of dry DMF was heated and dissolved. After cooling to room temperature, 5 g of 5, 9, 13, 17-tetramethyloctadecanoic acid chloride obtained in the synthesis of mono O- (5, 9, 13, 17-tetramethyloctadecanoyl) erythritol (11) was dissolved in 5 mL of dichloromethane and added dropwise. After the addition, it was stirred at room temperature for 1 hour. To the resulting reaction solution, 100 mL of dichloromethane was added, washed with saturated brine three times, and dried over anhydrous sodium sulfate. After filtration and concentration under reduced pressure, it was purified by silica gel column chromatography, whereby 2.50 g of mono O- (5, 9, 13, 17-tetramethyloctadecanoyl) pentaerythritol having the following physical properties was obtained. The purity of the product was 99.5% or more by HPLC analysis. In addition, the results of NMR measurement were as follows.

[0342] 1 H-NMR spectrum (270 MHz, CDC13, TMS) δ: 0.8-0.9 (m, 15H), 1.0-1.7 (m, 26H), 2.34 (t, J=7.4 Hz, 2H), 3.06 (br s, 3H), 3.63 (s, 6H), 4.17 (s, 2H)

[0343] Mono O- (5, 9, 13, 17-tetramethyloctadecanoyl) pentaerythritol is also referred to as saturated C22 pentaerythritol ester.

[0344] (14) Synthesis of mono O- (5, 9, 13, 17-tetramethyloctadec-4-enoyl) diglycerol

[0345] [Chemical Formula 22]

[0346]

[0347] To a solution of 259 g (1.56 mol) of diglycerol and 1.58 g (1.15 mmol) of potassium carbonate in dry N, N-dimethylformamide (700 mL), 199 g (0.564 mol) of 5, 9, 13, 17-tetramethyloctadec-4-enoic acid methyl ester was added dropwise at 78-83°C under reduced pressure of 60-70 mmHg and a stream of nitrogen. After stirring at the same temperature for 10 hours, formic acid was added at 75°C to adjust the pH to 4. The resulting solution was concentrated under reduced pressure, diluted with methyl tert-butyl ether (1.5 L), and the resulting insolubles were filtered off. The filtrate was washed with 10% aqueous sodium bicarbonate twice, treated with activated carbon (8 g), and decolorized. After filtration, the resulting residue was concentrated, dissolved in ethanol, and filtered through a cellulose powder. The concentrated residue was purified by silica gel column chromatography (mobile phase: hexane / ethyl acetate mixture), whereby the title compound was obtained as a transparent viscous liquid. With respect to the obtained compound, 1The results of the H-NMR measurement are shown below.

[0348] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.90 (m, 12H), 1.00-1.70 (m, 22H), 1.97 (ddd, J=6.9, 7.8, 17.4 Hz, 2H), 2.20-2.45 (m, 4H), 3.50-4.10 (m, 8H), 4.10-4.25 (m, 2H), 5.08 (dd, J=6.6, 6.6 Hz, IH)

[0349] Mono 0-(5,9,13,17-tetramethyloctadec-4-enoyl) diglycerol is also called C22 diglyceride.

[0350] (15) Synthesis of Mono 0-(3,7,11,15-tetramethylhexadec-2-enoyl) glycerol

[0351] [Chemical 23]

[0352]

[0353] To a solution of 0.57 g (6.2 mmol) of glycerol and 0.85 g (6.2 mmol) of potassium carbonate in dry N,N-dimethylformamide (3 mL), 1.0 g (3.1 mmol) of 3,7,11,15-tetramethylhexadec-2-enoic acid methyl ester was slowly dropped at 80°C. After stirring at 100°C for 12 hours, 1 M hydrochloric acid was added to the reaction solution, and extracted with ether. The extracted solution was washed with saturated sodium bicarbonate water, saturated brine in this order, dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to obtain 459 mg (yield 35%) of the title compound as a colorless sticky substance. With respect to the obtained compound, 1 The results of the H-NMR measurement are shown below.

[0354] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 1.00-1.80 (m, 19H), 1.90-2.20 (m, 5H), 3.50-4.00 (m, 3H), 4.10-4.30 (m, 2H), 5.71 (br s, IH)

[0355] Mono 0-(3,7,11,15-tetramethylhexadec-2-enoyl) glycerol is also called C20 glyceride, or phytoglycerol.

[0356] (16) Synthesis of mono O-(3,7,11,15-tetramethylhexadecanoyl) glycerol

[0357] [Chemical 24]

[0358]

[0359] To a solution of 2.52 g (6.10 mmol) of C20 glyceride in ethyl acetate (15 mL) was added 0.26 g of 5% palladium on carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred for 1 day at room temperature under an ordinary pressure hydrogen gas atmosphere. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (mobile phase: ethyl acetate) to thereby obtain the title compound 2.50 g (yield 99%) as a colorless transparent liquid. With respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0360] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.8-0.9 (m, 12H), 0.9-1.0 (m, 3H), 1.0-2.0 (m, 22H), 2.1-2.4 (m, 2H), 3.4-4.0 (m, 3H), 4.05-4.25 (m, 2H)

[0361] Mono O-(3,7,11,15-tetramethylhexadecanoyl) glycerol is also called saturated C20 glyceride, or phytanic acid glyceride.

[0362] [Example 2] Preparation of precursor formulation, gel formation test, and analysis of liquid crystal structure (1)

[0363] An amphiphilic compound having a isoprenoid-type aliphatic chain having a carbon number of 22 (isoprenoid-type lipid) synthesized in Example 1 as a self-organizing lipid (hereinafter also referred to as SOL, and abbreviated as SOL in the table), soybean phosphatidylcholine (LIPOID S100, Lipoid Co.; also abbreviated as SPC hereinafter and in the table) as a phospholipid, oil, and alcohol were mixed in the mixing ratio shown in Table 1 below. Note that the self-organizing lipid (SOL) is a lipid that forms a gel when mixed with water alone. Also, "EtOH" in the table is ethanol, and "PG" is propylene glycol (also the same in Tables 1 to 5, 8 to 12). In some of the formulations, P80 [polyoxyethylene sorbitan monooleate (20E.O.), NIKKOL TO-10MV, Nikko Chemicals Co.] as a surfactant and HPC (hydroxypropyl cellulose, HPC-SSL, Wako Pure Chemical Industries, Ltd.) as a water-soluble polymer were added. The resulting mixture was dissolved in a water bath at 40°C or lower, whereby Preparations 1 to 61 shown in Table 1 were prepared.

[0364] In addition, as a phospholipid, EPC (phosphatidylcholine from egg yolk, 163-21181, Fuji Photo Film Co. and Wako Pure Chemical Industries, Ltd.), DMPC (dimyristoyl phosphatidylcholine, COATSOME MC-4040, NOF Corporation), or DPPC (dipalmitoyl phosphatidylcholine, COATSOME MC-6060, NOF Corporation) was used instead of SPC, and Preparations 62 to 64 were prepared in the same manner as described above in the mixing ratio shown in Table 2.

[0365] In addition, an amphiphilic compound having a isoprenoid-type aliphatic chain having a carbon number of 17 synthesized in Example 1, or glycerol monooleate (Rikemar XO-100, NOF Corporation) having a straight-chain oleic acid as an aliphatic chain was used instead of the amphiphilic compound having a isoprenoid-type aliphatic chain having a carbon number of 22; or, no amphiphilic compound having a isoprenoid-type aliphatic chain was used, and Preparations 65 to 75 were prepared in the same manner as described above in the mixing ratio shown in Table 3.

[0366] The precursor preparations Nos. 1 to 75 thus obtained were subjected to a gel formation test. To a vial was added an excess of an aqueous medium [0.5 to 2 mL or so of water for injection for Nos. 1 to 61 and Nos. 64 to 75; 0.5 to 2 mL or so of a phosphate buffer physiological saline (PBS) having a pH of 7.4 for Nos. 62 and 63] and a portion (100 to 300 mg or so) of each of the precursor preparations, and mixed at room temperature (25°C) using a spatula and / or a vortex mixer. As a result, in all of the precursor preparations Nos. 1 to 75, a gel-like composition which was separated out in the excess of the aqueous medium and which was colorless transparent to white turbid in appearance was obtained.

[0367] The gel-like compositions obtained from the precursor preparations Nos. 1 to 70 were directly filled into a needle hole slit, and subjected to small angle X-ray scattering diffraction measurement using a small angle X-ray scattering (SAXS) device (Rigaku Corporation, Nano-Viewer) to analyze the non-lamellar liquid crystal structure.

[0368] The liquid crystal phase obtained and the scattering vector value ql [nm"1] of the peak on the side of the smallest angle are shown in Tables 1 to 3. Note that HII means a reverse hexagonal liquid crystal, Pn3m means a reverse cubic liquid crystal belonging to the crystal space group Pn3m, Im3m means a reverse cubic liquid crystal belonging to the crystal space group Im3m, and Fd3m means a reverse cubic liquid crystal belonging to the crystal space group Fd3m. Among the gel-like compositions, there are both gel compositions having two sizes (scattering vector values ql) in the same liquid crystal phase and gel compositions having two different liquid crystal phases.

[0369] As shown in Tables 1 and 2, when the SOL synthesized in Example 1, i.e., the amphiphilic compound having a isoprenoid-type aliphatic chain having 22 carbon atoms, was used, the precursor preparation containing the SOL was able to form a non-lamellar liquid crystal in a wide range of ratios of SOL:phospholipid (weight ratio) of 100:0 to at least 30:70, regardless of the kind of the amphiphilic compound, the phospholipid, and the oil. In addition, the precursor preparation containing the SOL was able to form a non-lamellar liquid crystal in a wide range of ratios of SOL + phospholipid (total amount of SOL and phospholipid):oil (weight ratio) of 100:0 to at least 20:80. Furthermore, even when P80 and HPC were added as additives, the precursor preparation containing the SOL formed a liquid crystal gel well. Obviously, various mixtures of the amphiphilic compound, the phospholipid, and the oil exhibited a variety of liquid crystal structures such as a reverse cubic liquid crystal belonging to the crystal space group Pn3m, Im3m, or Fd3m, and a reverse hexagonal liquid crystal, and the value of ql caused by the lattice constant could take a very wide range of values.

[0370] Note that when SPC was used as the phospholipid and sesame oil (triglyceride) was used as the oil, no liquid crystal gel was formed in the absence of SOL at a phospholipid: oil ratio of 85: 15 or 75:25 (by weight) (not shown in the table), and therefore when sesame oil (triglyceride) is used at these ratios, if the phospholipid is to be mixed to form a liquid crystal gel, it is necessary to add SOL or an amphiphilic compound having an isoprenoid-type fatty chain (isoprenoid lipid).

[0371] [Table 1]

[0372]

[0373]

[0374]

[0375]

[0376] * The oil used is shown after the weight ratio.

[0377] [Table 2]

[0378]

[0379] * The phospholipid used is shown after the weight ratio.

[0380] [Table 3]

[0381]

[0382]

[0383] * The oil used is shown after the weight ratio.

[0384] [Example 3] Preparation of emulsions (dispersions) and analysis of liquid crystal structure (1)

[0385] An oily solution was obtained by mixing, in the mixing ratios shown in Table 4 below, an amphiphilic compound having an isoprenoid-type fatty chain with 22 carbon atoms synthesized in Example 1 as a self-organizing lipid (abbreviated as SOL in the table), soybean phosphatidylcholine (LIPOID S100, Lipoid Co.; abbreviated as SPC in the table) as a phospholipid, an oil, and an alcohol. On the other hand, Pluronic F127 (Unilube (R)70DP-950B, Nippon Oil) and distilled water (Otsuki distilled water) to obtain a Pluronic aqueous solution. In some of the preparations, P80 as a surfactant was added to the above oily solution, or ascorbic acid and sodium sulfite as antioxidants were added to the Pluronic aqueous solution. After the oily solution and the Pluronic aqueous solution thus prepared were completely dissolved in a water bath at 30°C or less, respectively, they were mixed together at room temperature with stirring by a medicine spoon or a stirring rod to obtain a suspension. The suspension was further dispersed by a high-pressure homogenizer (StarBurst mini mo, manufactured by SUGINO MACHINE Co., Ltd.) to prepare white emulsions containing microparticles (Preparations Nos. 76 to 105). These emulsions were prepared in amounts of 7 to 30 g, respectively.

[0386] In addition, instead of the amphiphilic compound having a isoprenoid-type aliphatic chain of 22 carbon atoms, an amphiphilic compound having a isoprenoid-type aliphatic chain of 17 or 20 carbon atoms synthesized in Example 1, sorbitan monooleate having a straight-chain oleic acid as an aliphatic chain (NIKKOL SO-10V, Nikko Chemicals Co., Ltd.), or purified sorbitan monooleate (a substance obtained by removing oleic acid, sorbitan dioleate, and the like, which are low-polarity components, from NIKKOL SO-10V by silica gel column purification) was used; or, instead of SOL, emulsions of Nos. 106 to 116 were prepared in the same manner as described above in accordance with the mixing ratios shown in Table 5.

[0387] The emulsions of the preparations No. 76 to No. 116 thus obtained were subjected to structural analysis by small-angle X-ray scattering (SAXS) using a NANO Viewer nanometer-scale X-ray structure evaluation apparatus (manufactured by Rigaku Corporation). Each emulsion was introduced into a capillary under atmospheric pressure, and the measurement was performed in a state of reduced pressure in the apparatus (the sample itself was under atmospheric pressure). In the scattering intensity distribution obtained from the emulsions of the preparations No. 76, No. 77, No. 81 to No. 86, No. 88, No. 89, No. 92 to No. 103, No. 105 to No. 109, and No. 115, at least two scattering peaks were observed. The peak ratio showed a ratio of 1 :V3:2 characteristic of inverse hexagonal liquid crystal, and thus it was indicated that these emulsions were liquid crystal emulsions (hexosomes) in which microparticles of inverse hexagonal liquid crystal were dispersed in an aqueous phase. Among them, in the scattering intensity distribution obtained from the emulsions of the preparations No. 77, No. 83 to No. 86, No. 88, No. 89, No. 92, No. 103, and No. 105, both the scattering peak characteristic of inverse hexagonal liquid crystal and a broad scattering peak were observed, and thus it was considered that these emulsions were liquid crystal emulsions containing a sponge phase (L3 phase). Among them, in the scattering intensity distribution obtained from the emulsions of the preparations No. 79, No. 87, No. 90, No. 91, No. 104, and No. 110 to No. 114, a broad scattering peak was observed, and thus it was considered that these emulsions were liquid crystal emulsions in which microparticles of a sponge phase (L3 phase) were dispersed in an aqueous phase. In the scattering intensity distribution obtained from the emulsion of the preparation No. 78, at least three scattering peaks were observed, and the peak ratio showed a ratio of V2:V4:V6 characteristic of cubic liquid crystal belonging to crystal space group Im3m, and thus it was indicated that these emulsions were liquid crystal emulsions in which microparticles of cubic liquid crystal belonging to crystal space group Im3m were dispersed in an aqueous phase. In addition, in the scattering intensity distribution obtained from the emulsions of the preparations No. 80 and No. 116, at least six scattering peaks were observed, and the peak ratio showed a ratio of V3:V8:V11 :V12:V16:V19 characteristic of cubic liquid crystal belonging to crystal space group Fd3m, and thus it was indicated that these emulsions were liquid crystal emulsions in which microparticles of cubic liquid crystal belonging to crystal space group Fd3m were dispersed in an aqueous phase.

[0388] When the amphiphilic compound having a carbon number of 22 of the isoprenoid-type aliphatic chain synthesized in Example 1 was used as SOL, liquid crystal emulsions having a non-lamellar liquid crystal could be prepared in a wide range of SOL: phospholipid ratios and SOL + phospholipid: oil ratios shown in Table 4, as in the case of the liquid crystal gels and the flowable materials obtained from the precursor preparations shown in Example 2. In addition, even when a certain amount of P80 and an antioxidant were added as additives, the liquid crystal emulsions could be favorably prepared.

[0389] [Table 4]

[0390]

[0391] * The oil used is described at the back of the weight ratio.

[0392] [Table 5]

[0393]

[0394] The oil used is described below in terms of weight ratio.

[0395] [Example 4] Safety evaluation in vivo (1)

[0396] The precursors of the preparations No. 1 to 6, No. 8 to 10, No. 12 to 16, No. 19, No. 21 to 24, No. 27, No. 31, No. 34, No. 38, No. 50, No. 52 to 54, No. 57, No. 59, No. 61, No. 65 to 67, No. 69, and No. 71 prepared in Example 2, and the emulsions of the preparations No. 76 to 85, No. 88, No. 89, No. 91 to 93, No. 102, No. 103, No. 106, No. 108, and No. 109 prepared in Example 3 were evaluated for safety in vivo by intraperitoneal administration as described below. The evaluation was performed using 8- to 10-week-old female Wistar rats.

[0397] Note that the precursors of the preparations form a non-lamellar liquid crystal after intraperitoneal administration, and thus become a bulk gel state. Therefore, when a foreign body reaction is induced, the foreign body reaction-related phenomena (abscesses having white pus, etc.) can be observed at a size that can be directly visually recognized. Note that the abscesses generated by the present application are cystic tissues formed by the accumulation of leukocytes, etc. caused by a non-infectious sterile foreign body reaction. On the other hand, the emulsions are in a solution state containing non-lamellar liquid crystal microparticles, and thus can spread throughout the entire abdominal cavity. Therefore, when a tissue disorder is induced, the tissue disorder in the entire abdominal cavity can be observed. In addition, since the emulsions are microparticles, the absorption transfer to the blood through the peritoneum, etc. is faster than that of the bulk gel state, and thus systemic toxicity (mainly liver toxicity) can be easily and clearly observed.

[0398] (1) Safety evaluation of the precursors of the preparations

[0399] First, the rats were subjected to general anesthesia using pentobarbital, and placed in a supine position. An abdominal incision of about 30 mm was made in the middle of the abdomen, and an abdominal incision of about 20 mm was made in the upper abdominal wall. The peritoneum was completely hemostatic. The peritoneal incision was closed by continuous suture using 5-0 silk thread for 6 stitches. About 23 mg of the precursor of the preparation (using a pipette with a 30 μL scale) was added dropwise and spread along the incision. After the sample application site was sprayed with water for injection (Otsuka distilled water) using a manual simple spray bottle (Spray Vial No. 2, manufactured by Maruemu Corporation) (5 sprays, about 180 mg), the abdominal wall was immediately closed by double-layer suture, and the operation was completed.

[0400] Seven days after the surgery, general anesthesia was performed again, and laparotomy was performed. The abdominal cavity was observed to evaluate the safety of the precursor preparation applied.

[0401] Regardless of the number of carbon atoms, whether it was an isoprenoid-type aliphatic chain or a straight-chain aliphatic chain, abscesses in the form of a gel-enclosing abscess were confirmed in the precursor preparations of Formulations No. 1, 2, 9, 13, 14, 22, 52, 53, 65, 66, and 71, which did not contain phospholipids or contained a small amount of phospholipids. The abscesses were leukocyte infiltrations unrelated to infection, and were white abscesses resulting from a foreign body reaction against the precursor preparation applied. Examples of photographs showing the abscesses produced are shown in Figure 1 Figure 1 A to C: precursor preparation of Formulation No. 1 was applied; Figure 1 D: precursor preparation of Formulation No. 71 was applied. Figure 1 The arrows in A to D indicate abscesses.

[0402] On the other hand, in the precursor preparations of Formulations No. 3 to 6, 8, 10, 12, 15, 16, 19, 21, 23, 24, 27, 31, 34, 38, 50, 54, 57, 59, 61, 67, and 69, which contained a certain amount or more of phospholipids, no abdominal cavity conditions related to side effects, including abscesses, were confirmed.

[0403] In the precursor preparation of Formulation No. 65, which contained C17 glycerides and did not contain phospholipids, not only the above-mentioned abscesses but also slightly enlarged liver and adhesion of the liver to the omentum were observed.

[0404] The above results show that in the precursor preparations produced in Example 2, regardless of whether or not they contained oil, surfactant, and water-soluble polymer, as long as they contained a certain amount or more of phospholipids, no foreign body reaction or side effects caused by a foreign body reaction occurred. In particular, when C22 sorbitan ester or saturated C22 sorbitan ester was used, formulations containing at least 30% phospholipids in the SOL:phospholipid ratio completely did not produce side effects, including foreign body reactions.

[0405] Thus, it was confirmed that the addition of phospholipids to the amphiphilic compounds having isoprenoid-type aliphatic chains with a carbon number of 17 and 22 synthesized in Example 1 improved the safety of the precursor preparations thereof. In addition, in the case of formulations using amphiphilic compounds having isoprenoid-type aliphatic chains with a carbon number of 22, even if the amount used was higher, side effects were not easily produced.

[0406] (2) Safety evaluation of emulsions

[0407] ​First, the rats were anesthetized with pentobarbital, and placed in a supine position. An amount of the emulsion prepared in Example 3 was administered intraperitoneally using a syringe (TERUMO syringe 1 mL) having a 26G needle. Alternatively, if the emulsion was highly viscous and difficult to pass through the 26G needle, a needleless syringe (TERUMO syringe 1 mL) was used to administer an amount of the emulsion intraperitoneally from an approximately 1 cm incision made in the midline of the abdomen. After administration, the abdominal wall was closed with double-layer sutures, and the operation was completed.

[0408] Seven days after the operation, general anesthesia was again performed, and the abdomen was opened. The intraperitoneal condition was observed to evaluate the safety of the administered emulsion.

[0409] As a result, in the emulsions of Formulations No. 76, 78, 80, and 81, which are amphiphilic compounds having isoprenoid-type fatty chains having 22 carbon atoms, and which do not contain phospholipids or contain a small amount of phospholipids, side effects were observed. For example, in the emulsion of Formulation No. 76, which contains C22 glycerides and does not contain phospholipids, when the administration amount was 360 μL, a small amount of ascites was observed in 1 of 3 cases; when the administration amount was 720 μL, 1 of 3 cases died after 3 days, and the remaining 2 cases showed slight hepatomegaly. In addition, in the emulsions of Formulations No. 78, 80, and 81, which contain C22 pentaerythritol esters or C22 sorbitan esters as amphiphilic lipids having isoprenoid-type fatty chains having 22 carbon atoms, and which do not contain phospholipids or contain a small amount of phospholipids, the same degree of slight hepatomegaly and / or adhesion around the liver as in the emulsion of Formulation No. 76 using C22 glycerides was observed.

[0410] In the emulsions of Formulations No. 108 and No. 109 containing C20 glycerides or saturated C20 glycerides having isoprenoid-type fatty chains with 20 carbon atoms and not containing phospholipids, side effect-related results were also observed. For example, in the emulsion of Formulation No. 108 containing C20 glycerides and not containing phospholipids, 3 out of 3 cases died after 1 day at an administration amount of 220 μL. In the emulsion of Formulation No. 109 containing saturated C20 glycerides and not containing phospholipids, 1 out of 3 cases died after 3 days at an administration amount of 220 μL, and the remaining 2 cases observed various side effect-related results such as ascites, hepatomegaly, perihilar adhesion, and whitening of the liver surface. This indicates that the safety of the emulsion using C20 glycerides or saturated C20 glycerides is significantly lower even if phospholipids are mixed. On the other hand, in the emulsions of Formulations No. 77, No. 79, No. 82 to No. 85, No. 88, No. 89, No. 91 to No. 93, No. 102, and No. 103 containing amphiphilic compounds having isoprenoid-type fatty chains with 22 carbon atoms and containing phospholipids in an amount of at least 720 μL, no side effect-related results were observed in the entire abdominal cavity. For example, the SOL:phospholipid ratio in the emulsions of Formulations No. 82 and No. 102 was 70:30. Note that the total weight of the lipids (SOL and phospholipids) and oil contained in 720 μL of the emulsion was 0.18 g in Formulations No. 77, No. 82 to No. 85, No. 93, and No. 103; 0.15 g in Formulations No. 88, No. 89, No. 91, and No. 92; and 0.14 g in Formulations No. 79 and No. 102, which correspond to 60 g, 50 g, 48 g (assuming a rat weight of 150 g and a human weight of 50 kg), respectively, in terms of the administration amount to a human.

[0411] In the emulsion of Formulation No. 106 containing C17 glycerides having isoprenoid-type fatty chains with 17 carbon atoms and not containing phospholipids, side effect-related results were observed from an administration amount of 72 μL, and at an administration amount of 220 μL, 3 out of 3 cases observed hepatomegaly and perihilar adhesion, and 1 case observed a small amount of ascites; at an administration amount of 720 μL, 3 out of 3 cases died after 1 day. Thus, it was found that the emulsion using C17 glycerides had a stronger side effect on the liver than the emulsion using amphiphilic compounds having isoprenoid-type fatty chains with 22 carbon atoms when no phospholipids were contained.

[0412] The above results indicate that the safety of the emulsions prepared in Example 3 is improved as long as they contain a certain amount or more of phospholipids, regardless of whether they contain oil, surfactants, or antioxidants. This trend is particularly evident in emulsions using amphiphilic compounds with isoprene-type fatty chains having 22 carbon atoms, where they remain extremely safe even at high application rates. Emulsions using amphiphilic lipids with isoprene-type fatty chains having 20 carbon atoms do not offer any safety advantage in vivo compared to emulsions using amphiphilic compounds with isoprene-type fatty chains having 17 or 22 carbon atoms.

[0413] Furthermore, formulations that do not contain phospholipids but contain additives selected from the group consisting of oils, surfactants, antioxidants and water-soluble polymers (HPCs) exemplified in this embodiment do not show improved safety compared to formulations without these additives.

[0414] Reports (e.g., non-patent literature 2) indicate that when glyceryl monooleate is used as SOL, if a citrate fatty acid glyceride (e.g., Grindsted, manufactured by Danisco A / S, Denmark), known as a negatively charged surfactant, is added... (R) [Citrim LR10, BC-FS SG, etc. are commercially available], which would improve safety. However, when safety evaluations were conducted on emulsions containing C17 glycerides in formulation 106, replacing the C17 glycerides with a mixture of C17 glycerides:citric acid fatty acid glycerides at a ratio of 80:20, 60:40, or 40:60, the same side effects as those in the emulsion of formulation 106 were observed. Furthermore, when evaluating the safety of a prodrug formulation with a C17 glycerides:citric acid fatty acid glycerides ratio of 60:40, the same side effects as those in the phospholipid-free prodrug formulation 65 were observed. Therefore, it is shown that adding citrate fatty acid glycerides does not improve the safety of the formulations using amphiphilic lipids with isoprene-like fatty acid chains involved in this invention.

[0415] The above results indicate that, regardless of dosage form (e.g., prodrugs, emulsions), formulations containing a certain amount of phospholipids and amphiphilic compounds with isoprene-type fatty acid chains have high safety profiles. In particular, formulations containing amphiphilic compounds with isoprene-type fatty acid chains with 22 carbon atoms, while having a higher risk of side effects when phospholipids are absent, have been shown to be extremely safe as long as they contain a certain amount of phospholipids, and can be safely used in vivo even at high dosages.

[0416] [Example 5] Particle size and safety evaluation of emulsion (1)

[0417] The particle size distribution of the emulsions of Formulations No. 77, 79, 81 to 105, 107, and 110 to 116 containing phospholipids, which were prepared in Example 3, was measured by dynamic light scattering using a Zetasizer Nano-ZS (manufactured by Malvern Instruments Ltd.). The measurement sample was prepared by diluting each emulsion, which was prepared immediately after preparation (within 2 days at room temperature) or after a certain period at room temperature, 200-fold with distilled water. Table 6 shows the average particle size (nm) (Z-Average) obtained from each measurement sample, the elapsed period of the emulsion at room temperature from the day of preparation, and the change amount (nm) of the average particle size (average particle size of the emulsion after a certain period at room temperature - average particle size of the emulsion immediately after preparation).

[0418] With respect to the stability with time at room temperature, the emulsions of Formulations No. 83, 84, 86 to 88, 92, 94, 95, and 98 to 100 of the amphiphilic compound having an isoprenoid-type aliphatic chain having 22 carbon atoms, which were synthesized in Example 1, had almost no change in the average particle size after a long period of time. In addition, the appearance of these emulsions also had almost no change.

[0419] On the other hand, the emulsions of No. 110 to 114 using sorbitan monooleate (NIKKOL SO-10V) having a straight-chain oleic acid as the aliphatic chain or purified sorbitan monooleate had a large increase in the average particle size. In addition, the appearance of these emulsions became white and thick, and some samples apparently produced flocculation. Thus, the formulations containing sorbitan monooleate were not stable regardless of the purity of the sorbitan monooleate.

[0420] Therefore, the emulsion containing the amphiphilic compound having an isoprenoid-type aliphatic chain having 22 carbon atoms and phospholipids was apparently much more excellent in stability with time than the emulsion using the amphiphilic compound having a straight-chain oleic acid as the aliphatic chain and phospholipids.

[0421] [Table 6]

[0422]

[0423]

[0424] [Example 6] Confirmation of Spray Performance

[0425] Each emulsion of No. 77, 79, 81 to 84, 87 to 90, 92 to 99, and 101 to 105 preparations containing the amphiphilic compound having a isoprenoid-type aliphatic chain with 22 carbon atoms and phospholipid prepared in Example 3 was filled into a manual spray vial (Spray Vial No. 2, manufactured by Maruemu Co.) in an amount of 1 to 5 mL, whereby a pump spray was prepared.

[0426] Each emulsion of any of the above preparations was sprayed in a mist or stick shape, and sprayed once at a distance of about 2 cm from the test surface, and the range of emulsion adhesion was substantially circular.

[0427] [Example 7] Evaluation of the effectiveness of the anti-adhesion effect

[0428] The anti-adhesion effect of the emulsions of No. 31 and 72 to 75 precursor preparations, and No. 83 to 97, 99 to 101, 104, 105, 115, and 116 were evaluated using 10-week-old female Wistar rats.

[0429] First, the rats were subjected to general anesthesia using pentobarbital, and an abdominal incision of about 30 mm was made in the middle of the abdomen in a supine position. An incision of about 20 mm was made in the left and right upper abdominal wall parietal peritoneum, and hemostasis was performed. The left and right peritoneal incision portions were closed by continuous suture with 5-0 silk thread for 6 stitches.

[0430] Next, about 23 mg (using a pipette set to a 30 μL scale) of the precursor preparation was added dropwise and spread over the right peritoneal incision portion, or about 72 mg of the emulsion was added dropwise and spread, and the suture portion was covered (sample application side). When the precursor preparation was used, water for injection (Otsu distilled water) was sprayed (5 sprays, about 180 mg) onto the sample application site using a manual spray vial (Spray Vial No. 2, manufactured by Maruemu Co.) to induce the formation of liquid crystals. No sample was applied to the left peritoneal incision portion (non-application side / control group). The sample was applied to the right peritoneal incision portion, and the abdominal wall was closed (laparotomy) immediately after the suture portion was closed, and the operation was completed.

[0431] Seven days after the operation, general anesthesia was performed again, and the abdominal incision was performed. The adhesion of the incision and suture portions of the sample application side and the non-application side was evaluated according to the following definitions of adhesion strength score and adhesion range rate. Note that in this evaluation, no adverse effects were observed in any of the above preparations.

[0432] Adhesion strength:

[0433] • 0 No adhesion

[0434] • Grade 1 slight pulling can peel off adhesion (without tissue damage)

[0435] • Grade 2 strong pulling can peel off adhesion (without tissue damage)

[0436] • Grade 3 strong pulling to peel off adhesion causes tissue damage

[0437] Adhesion range rate:

[0438] Ratio (%) of adhesion length to about 20 mm of incision suture part

[0439] Determination:

[0440] Ratio of adhesion range of application sample incision part to non-application incision part (adhesion range rate of sample application side / adhesion range rate of non-application side x 100) is: 40% or less is A; 41% to 60% is B+; 61% to 80% is B-; 81% or more is C.

[0441] The evaluation results of the anti-adhesion effect are shown in Table 7.

[0442] The precursor preparations No. 31 containing both sorbitan ester and phospholipid as SOL and emulsions No. 83 to 97, 99 to 101, 104, and 105 exhibited high adhesion prevention (reduction) effect on the sample application side compared to the non-application side. On the other hand, the precursor preparations No. 72 to 75 and emulsions No. 115 and 116 containing no SOL exhibited low adhesion prevention (reduction) effect.

[0443] Therefore, the precursor preparations and emulsions containing both SOL and phospholipid had no side effects that could be confirmed in appearance in the abdominal cavity and apparently had adhesion prevention (reduction) effect.

[0444] [Table 7]

[0445]

[0446] In addition, using New Zealand white rabbits (female, 13 weeks old at the time of surgery) instead of Wistar rats, an evaluation test was performed on the anti-adhesion effect of the above non-lamellar liquid crystal preparation containing SOL and phospholipid. After the abdominal cavity of the rabbit was opened under anesthesia, the peritoneum and right internal oblique muscle on the right abdominal wall (side wall resection site) were excised, and the 6-lobed cecum was wiped with gauze (cecum wiping site), and dried under a incandescent lamp for about 60 minutes, to produce a cecum and peritoneum adhesion model. In the test group, 800 μL of the non-lamellar liquid crystal preparation was applied to each of the side wall resection site and the cecum wiping site, and the abdominal cavity was closed. As a control, in the physiological saline group, 5 mL of physiological saline was applied to each of the side wall resection site and the cecum wiping site, and the abdominal cavity was closed. The general state of each rabbit was observed once a day during the experiment period (on the day before and after the surgery), and the body weight was measured on the day of the surgery and the day of the autopsy. Seven days after the surgery, the rabbits were euthanized, the abdominal cavity was opened, and the preparation application site and the organs in the abdominal cavity were observed with the naked eye, and the adhesion at the preparation application site was evaluated. As in the case of using rats, an anti-adhesion effect was exhibited, and, for example, No. 88 preparation exhibited a higher anti-adhesion effect than the control physiological saline group. In the case of applying any of the preparations, no significant abnormalities were confirmed in the body weight progression of each animal, and no systemic toxicity was confirmed. In addition, no abnormal results (inflammation, hypertrophy, organ adhesion, etc.) were observed at the preparation application site and the organs in the abdominal cavity (liver, spleen, etc.), and no accumulation of ascites and pleural effusion was confirmed.

[0447] [Example 8] In vitro release test of leuprolide acetate (1)

[0448] In the same manner as in Example 2, C17 glyceride having an isoprenoid-type aliphatic chain with a carbon number of 17 (hereinafter referred to as SOL) synthesized in Example 1, surfactant P80 [polyoxyethylene sorbitan monooleate (20 E.O.)], and ethanol were mixed at the mixing ratio shown in Table 8 below, and stirred at 60°C for 5 minutes. To the resulting solution, DMPC (dimyristoyl phosphatidylcholine, COATSOME MC-4040, NOF Corporation), DOPC (dioleoyl phosphatidylcholine, COATSOME MC-8181, NOF Corporation), DOPE (dioleoyl phosphatidylethanolamine, COATSOME ME-8181, NOF Corporation), or DOPG-Na (dioleoyl phosphatidylglycerol sodium, COATSOME MG-8181LS, NOF Corporation) was added as a phospholipid, and stirred at 60°C for 1 hour, to thereby prepare Nos. 117 to 120 of the precursor preparation.

[0449] The precursor preparations No. 117 to 120 were added to a large excess of PBS at pH 7.4, and gellike compositions were obtained in all cases. Small-angle X-ray scattering measurements were performed on each of the gellike compositions in the same manner as in Example 2, and it was confirmed that all of the gellike compositions had a non-lamellar liquid crystal structure. The liquid crystal phase obtained and the value of the scattering vector q1[nm"1] of the peak on the side of the smallest angle are shown in Table 8.

[0450] [Table 8]

[0451]

[0452] * The phospholipids used are described below in terms of weight ratio.

[0453] Next, the precursor preparations No. 121 to 124 containing leuprolide acetate (prepared from No. 117 to 120, respectively) were prepared as follows. First, 3.75 mg of leuprolide acetate (L0249, Tokyo Chemical Industry Co., Ltd.) was added to a dialysis tube (Pur-A-Lyzer MINI 12000, Sigma-Aldrich), and after dissolution with 5 mg of dimethyl sulfoxide, 91.25 mg of the precursor preparations No. 117 to 120 were added. Each of the mixtures obtained was stirred with a Pellet Pestle for 2 minutes, and 100 mg of the precursor preparations No. 121 to 124 containing leuprolide acetate were thus obtained. In addition, as a comparative control, 3.75 mg of leuprolide acetate was added to a dialysis tube, and dissolved with 96.25 mg of PBS at pH 7.4, and 100 mg of an aqueous solution No. 125 containing leuprolide acetate was thus obtained. TM MINI 12000, Sigma-Aldrich), and after dissolution with 5 mg of dimethyl sulfoxide, 91.25 mg of the precursor preparations No. 117 to 120 were added. Each of the mixtures obtained was stirred with a Pellet Pestle for 2 minutes, and 100 mg of the precursor preparations No. 121 to 124 containing leuprolide acetate were thus obtained. In addition, as a comparative control, 3.75 mg of leuprolide acetate was added to a dialysis tube, and dissolved with 96.25 mg of PBS at pH 7.4, and 100 mg of an aqueous solution No. 125 containing leuprolide acetate was thus obtained.

[0454] In vitro release tests were performed on the precursor preparations No. 121 to 124 containing leuprolide acetate (100 mg) and the aqueous solution No. 125 (100 mg). For a vial (size 25 mL) filled with 20 mL of a PBS solution at pH 7.4 containing 0.02% P80, a dialysis tube was attached to the upper portion with a float, so that the precursor preparation placed inside was sufficiently immersed in the PBS solution, and left standing at room temperature (25°C). Next, from the PBS solution in the vial, 500 μL was collected at 0, 1, 3, 6, 12, 24, 48, 72, 120, and 168 hours after the start of the test over a period of 7 days. Note that for the precursor preparations No. 121 to 124 added to the PBS solution described above, a change to gellike compositions was observed by visual observation.

[0455] As for the quantification of leuprolide acetate in the samples collected from the PBS solution, analysis by LC / MS / MS was performed using a calibration curve drawn in advance. The analysis conditions were as follows.

[0456] • Analysis column: Shodex ODP2HP-2B 2.0 mm I.D. x 50 mm (Showa Denko K.K.)

[0457] • Mobile phase: water (containing 0.1% formic acid) / acetonitrile = 70:30

[0458] • Flow rate: 0.1 mL; column temperature: 40°C; injection volume: 10 μL

[0459] • Precursor ion: 605.3 m / z; product ion: 249.0 m / z

[0460] Figure 2 In vitro release data from precursor preparations No. 121 to 124 containing leuprolide acetate and No. 125 aqueous solution are shown. Figure 2 In the case of No. 125 aqueous solution, all of the leuprolide acetate was rapidly released within 1 or 2 days, whereas in the case of No. 121 to 124 precursor preparations, no initial burst was observed, indicating sustained release of leuprolide acetate. In this regard, the No. 122 precursor preparation showed slightly faster release, the No. 124 precursor preparation showed slightly slower release, and so on, indicating that the release rate of leuprolide acetate can be controlled by changing the type of phospholipid. This result indicates that the preparations of the present application, represented by No. 121 to 124 precursor preparations, can be used as long-acting agents.

[0461] [Example 9] Subcutaneous implantation test (1)

[0462] Three types of mixed anesthesia (metomidate hydrochloride + midazolam + butorphanol tartrate) were used to anesthetize 7- to 8-week-old male Wistar rats, and after the back was shaved, 100 μL of the precursor preparations (No. 117 and 118, containing C17 glyceride and DMPC or DOPC as phospholipid) prepared in the same manner as in Example 8 were subcutaneously administered to the back using a syringe (TERUMO syringe 1 mL) having a 23G needle.

[0463] After 2 days from the administration, general anesthesia was performed again, and the skin containing the subcutaneous administration site of the preparation was removed. After the subcutaneous fat was simply removed, the administration site of the preparation was observed macroscopically.

[0464] Figure 3 Photographs of the observed administration sites of the preparations are shown in Figure 3 A: No. 117 precursor preparation; Figure 3B: Preparation of No. 118 precursor). None of the applied preparations were discolored in the site of the preparation application, and existed as a gel-like composition. In addition, no results caused by inflammation due to the preparation were observed in the surrounding tissue of the site of the preparation application in all rats to which the preparation was applied, and no abnormal results such as bleeding, capsule formation, discoloration, etc. were observed.

[0465] [Example 10] Synthesis of amphiphilic compound (2)

[0466] (1) Synthesis of mono O- (5, 9, 13-trimethyltetradec-4-enoyl) propylene glycol

[0467] [Formula 25]

[0468]

[0469] To a solution of 24.3 g (317 mmol) of propylene glycol and 0.30 g (2.2 mmol) of potassium carbonate in dry N, N-dimethylformamide (70 mL), 28.2 g (100 mmol) of 5, 9, 13-trimethyltetradec-4-enoic acid methyl ester was slowly added dropwise at 85°C under reduced pressure of 60 to 70 mmHg and a stream of nitrogen, and stirred for 5 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 160 mL), washed with water, saturated sodium bicarbonate aqueous solution, saturated brine (2 times), and 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 70:30), whereby 26.1 g of the title compound was obtained as a light yellow transparent liquid (yield 80%). With respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0470] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.75-0.90 (m, 9H), 0.95-1.70 (m, 18H), 1.97 (td, J = 7.6, 18 Hz, 2H), 2.07 (br s, OH), 2.25-2.42 (m, 4H), 3.55-3.70 (m, 0.7H), 3.85-4.15 (m, 1.95H), 4.98 (m, 0.35H), 5.08 (t, J = 6.2 Hz, IH)

[0471] Mono O- (5, 9, 13-trimethyltetradec-4-enoyl) propylene glycol is also called C17 propylene glycol ester.

[0472] (2) Synthesis of mono O- (5, 9, 13, 17-tetramethyloctadec-4-enoyl) propylene glycol

[0473] [Chem. 26]

[0474]

[0475] To a solution of 20.6 g (271 mmol) of propylene glycol and 0.211 g (1.53 mmol) of potassium carbonate in dry N,N-dimethylformamide (60 mL) was added 30.0 g (85.0 mmol) of methyl 5,9,13,17-tetramethyloctadeca-4-enoate slowly dropwise at 85°C under reduced pressure of 60 to 70 mmHg and a stream of nitrogen, and stirred for 3 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 120 mL), washed with water, saturated sodium bicarbonate aqueous solution, saturated brine (2 times), and dried over magnesium sulfate. The residue obtained by concentration after filtration was purified by silica gel column chromatography (mobile phase: hexane / ethyl acetate = 100:0 to 70:30), whereby 21.6 g of the title compound was obtained as a light yellow transparent liquid (yield 63%). With respect to the obtained compound, 1 The measurement results of1H-NMR are shown below.

[0476] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 0.95-1.70 (m, 25H), 1.96 (td, J = 7.5, 18 Hz, 2H), 2.1 (br s, OH), 2.25-2.42 (m, 4H), 3.55-3.70 (m, 0.7H), 3.85-4.15 (m, 1.95H), 4.98 (m, 0.35H), 5.08 (t, J = 6.8 Hz, IH)

[0477] Mono 0-(5,9,13,17-tetramethyloctadecanoyl) propylene glycol is also called C22 propylene glycol ester.

[0478] (3) Synthesis of Mono 0-(5,9,13,17-tetramethyloctadecanoyl) propylene glycol

[0479] [Chem. 27]

[0480]

[0481] To a solution of 3.96 g (10.0 mmol) of mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)propanediol in ethyl acetate (12 mL) was added 0.48 g of 5% palladium on carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, the system was stirred for 48 hours at room temperature under an ordinary pressure hydrogen gas atmosphere. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (ethyl acetate) to thereby obtain the title compound 3.94 g (yield 99%) as a colorless transparent liquid. The obtained compound was identified by1H-NMR. 1 The measurement results of1H-NMR are shown below.

[0482] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.75-0.9 (m, 15H), 0.95-1.7 (m, 29H), 2.28 (br s, OH), 2.30 (m, 2H), 3.54-3.67 (m, 0.62H), 3.88-4.1 (m, 2.07H), 4.97 (m, 0.31H)

[0483] Mono-O-(5,9,13,17-tetramethyloctadecanoyl)propanediol is also called saturated C22 propanediol ester.

[0484] [Example 11] Preparation of precursor formulations, gel formation test, and analysis of liquid crystal structure (2)

[0485] The isoprenoid-type lipid, i.e., the amphiphilic compound, synthesized in Example 1 or 10, SPC, DMPC, DPPC, DOPC, DOPE, DOPG-Na (all of which are the same products as those used in Example 2 or 8), or EPC (purified egg yolk lecithin, PL-100M, Nippon Emulsion Co., Ltd.), oil, and, as the case required, an alcohol were mixed in the mixing ratios shown in Table 9 below. The resulting mixture was dissolved in a water bath at 40°C or lower to thereby prepare precursor formulations Nos. 126 to 152 shown in Table 9. Of the isoprenoid-type lipids used, C17 propanediol ester and C22 propanediol ester are low-viscosity liquids, and precursor formulations (e.g., No. 147) can be prepared even without mixing an alcohol.

[0486] Note that C17 propanediol ester, C22 propanediol ester, and saturated C22 propanediol ester are not self-organizing lipids (SOLs) but isoprenoid-type lipids because they do not form gels when mixed with water alone.

[0487] Gel formation test was performed for the precursor preparations No. 126 to 152. To a vial, an excess of water for injection (0.5 to 2 mL or so) was added with a portion (100 to 300 mg or so) of each of the precursor preparations, and mixing operation was performed at room temperature (25°C) using a spatula and / or a vortex mixer. As a result, in all of the precursor preparations No. 126 to 152, a gel-like composition which was colorless transparent to white turbid in appearance was obtained which was separated in the excess of water for injection (aqueous medium).

[0488] With respect to the gel-like compositions obtained from the precursor preparations No. 126 to 152, non-lamellar liquid crystal structure analysis was performed by small-angle X-ray scattering diffraction using the same method as in Example 2, and the liquid crystal phase and the scattering vector value ql [nm"1] of the peak on the side of the smallest angle were determined. As shown in Table 9, not only the isoprenoid lipid of the self-organizing lipid (SOL), but also the precursor preparations containing the combination of the isoprenoid lipid and the phospholipid formed a non-lamellar liquid crystal in the presence of water when using the isoprenoid lipid other than the SOL.

[0489] Note that, when the same precursor preparations as above were prepared using a pharmaceutical additive propylene glycol monostearate (NIKKOL PMS-1CV, Kanto Chemical Co., Inc.) having a straight-chain stearic acid as a fatty chain instead of the above isoprenoid lipid, no liquid crystal gel was formed in the entire range of the propylene glycol monostearate: phospholipid ratio (weight ratio) of 100:0 to 0:100.

[0490] [Table 9]

[0491]

[0492] * The phospholipids used are described below in the weight ratio.

[0493] [Example 12] Preparation of emulsions (dispersions) and analysis of liquid crystal structure (2)

[0494] Using the C17 glyceride and the C22 propylene glycol ester synthesized in Examples 1 and 10, respectively, white emulsions 153 and 154 (8 g each) containing microparticles were prepared in the mixing ratios shown in Table 10 below using the same method as in Example 3.

[0495] For the emulsions of Formulations No. 153 and 154, structural analysis was performed using small angle X-ray scattering (SAXS) using the same method as in Example 3. In the scattering intensity distribution obtained from the emulsion according to the emulsion of Formulation No. 153, a broad scattering peak was observed, and it was thus considered that the emulsion was a liquid crystal emulsion in which microparticles of a sponge phase (L3 phase) were dispersed in an aqueous phase. In the scattering intensity distribution obtained from the emulsion according to the emulsion of Formulation No. 154, at least three scattering peaks were observed (the scattering vector value q1 of the peak on the side of the smallest angle was 1.08 nm"1). The ratio of the peaks showed a ratio specific to a reverse hexagonal liquid crystal of 1 :V3 : 2, and thus it was indicated that the emulsion was a liquid crystal emulsion in which microparticles of a reverse hexagonal liquid crystal were dispersed in an aqueous phase (hexosomes).

[0496] [Table 10]

[0497]

[0498] * The oils used are described below in terms of weight ratio.

[0499] [Example 13] Evaluation of safety in vivo (2)

[0500] For the precursor formulations of Formulations No. 127, 128, 130, 131, 134, 136, 137, 139, 143, 145, 146, and 152 prepared in Example 11, and the emulsions of Formulations No. 153 and 154 prepared in Example 12, evaluation of safety in vivo was performed by intraperitoneal administration using the same method as in Example 4.

[0501] (1) Results of evaluation of safety of precursor formulations

[0502] In the precursor formulations of Formulations No. 127 and 139, which had a small amount of phospholipid, abscesses were confirmed to have been generated in the form of a gel in which the surroundings were formed. On the other hand, in the precursor formulations of Formulations No. 128, 130, 131, 134, 136, 137, 143, 145, 146, and 152, which had a larger amount of phospholipid, no intraperitoneal conditions related to side effects, including abscesses as a foreign body reaction, were confirmed.

[0503] The components of the precursor formulation of Formulation No. 127, in which abscesses were generated, corresponded to components in which 62.5% of the amount of phospholipid contained in Formulation No. 15, in which no abscesses were generated in Example 4, was replaced with oil [isoprenoid lipid (SOL in Table 1): phospholipid: oil = 60: 15: 25]; in addition, the isoprenoid lipid (SOL in Table 1): phospholipid ratio was 80:20, which was the same as Formulation No. 14, in which abscesses were generated in Example 4. Thus, this result again indicated that, in order to ensure safety in vivo, not oil, but a certain amount or more of phospholipid was required relative to isoprenoid lipid.

[0504] In the intraperitoneal administration requiring higher safety than the subcutaneous implantation (Example 9), no foreign body reaction and other side effects occurred regardless of the kind of phospholipid. This result is crucial especially in the safety evaluation of the 131st, 134th, 136th, and 137th preparations in which the SPC of the 55th preparation was replaced with other various phospholipids (EPC, DMPC, DOPC, or DOPE).

[0505] It was also shown that, as the R of the general formula (I) is a hydrophilic group of the isoprenoid lipid having one hydroxyl group, not only in the case of the isoprenoid lipid from isosorbide but also in the case of the isoprenoid lipid from propylene glycol, no foreign body reaction and other side effects occurred as long as it was combined with an amount of phospholipid or more (see the 143rd, 145th, 146th, and 152nd preparations).

[0506] As to the isoprenoid lipid from propylene glycol ester, like the lipid from glycerol ester, the isoprenoid lipid having a carbon atom number of 22 is less likely to cause side effects than the isoprenoid lipid having a carbon atom number of 17 (the 139th preparation compared with the 145th preparation).

[0507] (2) Safety evaluation results of emulsions

[0508] The emulsion of the 153rd preparation using C17 glycerol ester and having a phospholipid ratio of up to 60% in the isoprenoid lipid:phospholipid ratio was administered to rats at an administration amount of 43 μL, and as a result, no results related to side effects were observed in the entire peritoneal cavity. Note that assuming that the rat weighs 150 g and the human weighs 50 kg, the administration amount per unit weight is equal, and thus the total weight of the isoprenoid lipid, phospholipid, and oil contained in the administration amount of 43 μL is converted to an administration amount for a human of 3.6 g. Among them, as to the 153rd preparation, slight hepatomegaly occurred at a larger administration amount of 360 μL (same as 30 g), and the degree of hepatomegaly increased at an administration amount of 720 μL (same as 60 g). Thus, it was shown that the emulsion containing the amphiphilic compound having an isoprenoid fatty chain having a carbon atom number of 17 and containing an amount of phospholipid or more can be safely administered in the peritoneal cavity at a smaller administration amount than the emulsion containing the amphiphilic compound having an isoprenoid fatty chain having a carbon atom number of 22 and containing an amount of phospholipid or more.

[0509] The emulsion of the No. 154 preparation using C22 propylene glycol ester, like other emulsions containing an amphiphilic compound having a isoprenoid-type aliphatic chain with the number of carbon atoms of 22 and containing an amount of phospholipid above the level shown in Example 4 (e.g., Nos. 77, 79, 82 to 85, 88, 89, 91 to 93, 102, and 103 preparations), also did not show results related to side effects in the entire abdominal cavity at an administration amount of 720 μL (the value converted to the human administration amount is 60 g).

[0510] [Example 14] In vitro release test of FD-4

[0511] The C22 glyceride having a isoprenoid-type aliphatic chain with the number of carbon atoms of 22 synthesized in Example 1, the surfactant P80 [polyoxyethylene sorbitan monooleate (20 E.O.)], and ethanol were mixed in the mixing ratio shown in Table 11 below, and stirred at 60°C for 5 minutes, in the same manner as in Example 2. To the obtained solution, DMPC, DPPC, or DOPC (all are the same products as those used in Example 2 or 8) was added as a phospholipid, and stirred at 60°C for 1 hour, whereby the precursor preparations of Nos. 155 to 158 were prepared.

[0512] The precursor preparations of Nos. 155 to 158 were added to a large excess of PBS at pH 7.4, and gelled compositions were obtained in all cases. Small-angle X-ray scattering measurement was performed on each gelled composition in the same manner as in Example 2, whereby it was confirmed that all the gelled compositions had a non-lamellar liquid crystal structure. The liquid crystal phase obtained and the scattering vector value q1 [nm"1] of the peak on the side of the smallest angle are shown in Table 11.

[0513] [Table 11]

[0514]

[0515] * The phospholipid used is shown in the weight ratio below.

[0516] Next, to the precursor preparations of Nos. 155 to 158, FD-4 (fluorescein isothiocyanate-dextran, average molecular weight 4,000, Sigma Aldrich, product number 46944) was added so as to make the preparation 15.6 mg / mL, and stirred for 3 minutes with a PelletPestle, whereby the precursor preparations of Nos. 159 to 162 containing FD-4 were obtained. In addition, as a comparative control, only the C22 glyceride was added and FD-4 was dissolved so as to make the preparation 15.6 mg / mL, whereby the precursor preparation of No. 163 of the C22 glyceride containing FD-4 and not containing a phospholipid was obtained.

[0517] Release test in vitro was performed for the precursors 159 to 163 containing FD-4. A dialysis tube (Pur-A-Lyzer MINI 12000, Sigma-Aldrich) was hydrated in advance with PBS, and 100 μL of the preparation was added thereto. For a vial (25 mL in size) filled with 20 mL of a PBS solution containing 0.01% sodium azide at pH 7.4, a dialysis tube was attached to the upper portion with a float to allow the precursor preparation placed inside to be sufficiently immersed in the PBS solution, and left to stand at room temperature (25°C). Subsequently, 500 μL was collected from the PBS solution in the vial at a time (an equal amount of PBS was added each time) over time from the start of the test until 48 hours later. It should be noted that for the precursors 159 to 163 added to the PBS solution, a change to a gel-like composition was visually observed. TM MINI 12000, Sigma-Aldrich) was hydrated in advance with PBS, and 100 μL of the preparation was added thereto. For a vial (25 mL in size) filled with 20 mL of a PBS solution containing 0.01% sodium azide at pH 7.4, a dialysis tube was attached to the upper portion with a float to allow the precursor preparation placed inside to be sufficiently immersed in the PBS solution, and left to stand at room temperature (25°C). Subsequently, 500 μL was collected from the PBS solution in the vial at a time (an equal amount of PBS was added each time) over time from the start of the test until 48 hours later. It should be noted that for the precursors 159 to 163 added to the PBS solution, a change to a gel-like composition was visually observed.

[0518] Quantification of FD-4 in the samples collected from the PBS solution was performed using a fluorescence spectrophotometer (RF-5300PC, Shimadzu Corporation, Japan) (excitation wavelength: 490 nm; fluorescence wavelength: 515 nm).

[0519] Figure 4 A graph showing the release in vitro from the precursors 159 to 163 containing FD-4 is shown. Figure 4 In the graph, the horizontal axis represents time [hours], and the vertical axis represents the release rate [%] of the amount of FD-4 released into the PBS solution at each time point relative to the amount of FD-4 contained in the precursor preparation at the start of the test (average value of n = 3). The release rate of FD-4 48 hours after the start of the test was 1% for the precursor 159, 35% for the precursor 160, 24% for the precursor 161, 39% for the precursor 162, and 1% for the precursor 163.

[0520] None of the precursors containing FD-4 showed an initial burst, but rather, the release of FD-4 started slowly. The precursors 163 not containing phospholipid and the precursor 159 containing a small amount of phospholipid released FD-4 slowly, while the precursors 160 to 162 containing a large amount of phospholipid released FD-4 more quickly.

[0521] These results indicate that the precursors in which various phospholipids were added to the C22 glyceride having a isoprenoid-type aliphatic chain having 22 carbon atoms as the isoprenoid-type lipid can control the release of the drug depending on the kind and amount of the phospholipid.

[0522] [Example 15] Release test in vitro of leuprolide acetate (2)

[0523] The same as in Example 2, C17 glyceride having a C17 isoprenoid fatty chain with a carbon number of 17, C22 propylene glycol ester [R in General Formula (I) has one hydroxyl group] having a C22 isoprenoid fatty chain with a carbon number of 22, C22 glyceride [R in General Formula (I) has two hydroxyl groups], C22 sorbitan ester component [R in General Formula (I) has three hydroxyl groups], or saturated C22 sorbitan ester component [R in General Formula (I) has three hydroxyl groups] as isoprenoid lipids, soybean phosphatidylcholine (LIPOID S100, Lipoid Co.; abbreviated as SPC in the table) as a phospholipid, surfactant P80 [polyoxyethylene sorbitan monooleate (20E.O.)], and ethanol were used in the mixing ratios shown in Table 12 below, and thus, No. 164 to No. 169 precursor preparations were prepared.

[0524] When No. 164 to No. 169 precursor preparations were added to PBS at pH 7.4 in a large excess amount, each resulted in a gel-like composition. Small-angle X-ray scattering measurement was performed on each of the gel-like compositions in the same manner as in Example 2, and thus, it was confirmed that all of the gel-like compositions had a non-lamellar liquid crystal structure. The liquid crystal phase obtained and the scattering vector value ql [nm"1] of the peak on the side of the smallest angle are shown in Table 12.

[0525] [Table 12]

[0526]

[0527]

[0528] Next, using the same method as in Example 8, No. 170 to No. 175 precursor preparations (100 mg) containing leuprolide acetate were prepared by adding each of No. 164 to No. 169 precursor preparations to a dimethyl sulfoxide solution of leuprolide acetate, and an in vitro release test of leuprolide acetate was performed for 7 days. Note that for No. 170 to No. 175 precursor preparations added to a PBS solution, each was visually observed to change to a gel-like composition.

[0529] Figure 5 In vitro release data from No. 170 to No. 175 precursor preparations containing leuprolide acetate are shown. Figure 5 In the graph, the horizontal axis represents time [days], and the vertical axis represents the release rate [%] of the amount of leuprolide acetate released into a PBS solution at each time point with respect to the amount of leuprolide acetate contained in the precursor preparation at the start of the test (average value of n = 3). The release rate of leuprolide acetate 7 days after the start of the test was 62% for No. 170, 7% for No. 171, 64% for No. 172, 45% for No. 173, 69% for No. 174, and 16% for No. 175.

[0530] The precursor formulations containing leuprolide acetate did not show initial burst, but slowly started to release leuprolide acetate. Among the precursor formulations Nos. 170 to 174, which were identical except for the isoprenoid lipid, the release rate of the precursor formulation No. 171 containing the C22 propylene glycol ester having the highest hydrophobicity was slow, and the release rates of the precursor formulations Nos. 170 and 172 to 174 containing the C17 glyceride, C22 glyceride, C22 sorbitan ester component, and saturated C22 sorbitan ester component, respectively, having similar HLB values were fast. In addition, the formulation No. 165 corresponding to the formulation No. 171 formed reverse hexagonal liquid crystals by the addition of PBS; the formulations Nos. 164 and 168 corresponding to the formulations Nos. 170 and 174 formed Pn3m cubic liquid crystals by the addition of PBS; the formulations Nos. 166 and 167 corresponding to the formulations Nos. 172 and 173 formed Pn3m cubic liquid crystals and reverse hexagonal liquid crystals by the addition of PBS. In particular, the release rate of the precursor formulation No. 174 containing the saturated C22 sorbitan ester component having a saturated aliphatic chain was the fastest. In addition, the release rate of the precursor formulation No. 175 containing the saturated C22 sorbitan ester component was slower than that of the precursor formulation No. 174 having a higher phospholipid ratio. The formulation No. 169 corresponding to the formulation No. 175 formed reverse hexagonal liquid crystals by the addition of PBS.

[0531] These results indicate that in the precursor formulations in which the isoprenoid lipid represented by the general formula (I) is added with the phospholipid, the release of the drug can be controlled depending on the type of the isoprenoid lipid and the content of the phospholipid.

[0532] [Example 16] Subcutaneous implantation test (2)

[0533] To 7-week-old male Sprague-Dawley (SD) rats [Crl:CD (SD), SPF], 72 μL of the emulsion of the formulation No. 87 prepared in Example 3, and 100 μL of any one of the 10 kinds of precursor formulations consisting of the formulations Nos. 35, 129, 145, 146, 3, 126, 23, 24, 54, and 56 prepared in Examples 2 and 11 were subcutaneously administered between the shoulder blades using a syringe equipped with a 22G injection needle (n = 3 for each formulation).

[0534] After 7 days from the administration, the skin containing the subcutaneous formulation administration site was taken out after the rats were bled under inhalation anesthesia with sevoflurane (Mylan Pharmaceuticals) and euthanized. After the subcutaneous fat was simply removed, the formulation administration site was observed macroscopically.

[0535] Figure 6 and Figure 7 Photographs showing the observed formulation administration sites Figure 6 A: Emulsion of the formulation No. 87; Figure 6 B: Precursor formulation No. 35;Figure 6 C: No. 129 precursor preparation Figure 6 D: No. 145 precursor preparation

[0536] Figure 6 E: No. 146 precursor preparation Figure 6 F: No. 3 precursor preparation Figure 7 A: No. 126 precursor preparation

[0537] Figure 7 B: No. 23 precursor preparation Figure 7 C: No. 24 precursor preparation Figure 7 D: No. 54 precursor preparation Figure 7 E: No. 56 precursor preparation

[0538] In the surrounding tissue of the preparation administration site of all rats to which the preparation was administered, no results caused by inflammation due to the preparation were observed, nor were abnormal results such as bleeding, discoloration, etc. observed.

[0539] At the administration site of the emulsion of No. 87 preparation, no preparation remained. At the administration site of No. 35 precursor preparation, No. 145 precursor preparation, No. 146 precursor preparation, No. 23 precursor preparation, No. 24 precursor preparation, No. 54 precursor preparation, No. 56 precursor preparation, which use C22 propylene glycol ester, C22 sorbitan ester component, or saturated C22 sorbitan ester component, among the above-mentioned 10 precursor preparations, there was a gel-like composition in the same volume as or less than the amount of preparation administered, in the same state as the gel-like composition observed at the time of the gel formation test. On the other hand, at the administration site of No. 129 precursor preparation, No. 3 precursor preparation, No. 126 precursor preparation, which use C17 glyceride, C22 glyceride, there was a gel-like composition in a larger volume than the above-mentioned gel composition, showing that the biological component infiltrated the composition.

[0540] The above results show that preparations (e.g., precursor preparations, emulsions) containing isoprenoid-type lipids typified by C17 glyceride, C22 propylene glycol ester, C22 glyceride, C22 sorbitan ester component, and saturated C22 sorbitan ester component, and phospholipids typified by SPC, are safe to use, as are the results of the safety evaluation by intraperitoneal administration (Examples 4, 13), when administered subcutaneously.

[0541] [Example 17] Synthesis of amphiphilic compounds (3)

[0542] (1) Synthesis of mono O-(3,7,11,15-tetramethylhexadec-2-enoyl) sorbitol

[0543] [Formula 28]

[0544]

[0545] After 6.49 g (20 mmol) of 3,7,11,15-tetramethylhexadec-2- enoic acid methyl ester and 5.47 g (30.0 mmol) of 90 wt% aqueous sorbitol (Sorbitol M-90, Sanoh Chemical Industry Co., Ltd., containing 90% solid content and 10% moisture, content in solid content: sorbitan 79-84%, isosorbide 15-18%) were added to a reaction vessel at room temperature, stirring was performed at 120°C, 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 0.22 g (4.0 mmol) of sodium methoxide and 4 mg of sodium hypophosphite monohydrate were added, and stirring was performed at 160°C, 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 0.11 g (2.0 mmol) of sodium methoxide was added, and stirring was performed at 160°C, 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 0.11 g (2.0 mmol) of sodium methoxide was further added, and stirring was performed at 160°C, 8 kPa for 1.5 hours. After cooling to 60°C, 20 mL of ethyl acetate and 20 mL of 0.5 M hydrochloric acid were added while stirring. To the resulting reaction solution, 60 mL of ethyl acetate was added, and extraction was performed. The extracted solution was washed with saturated sodium bicarbonate water, saturated brine, dried with magnesium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to obtain 3.83 g (yield 42%) of a component containing the title compound as a light brown transparent liquid. The resulting component contained mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl)sorbitan and mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl)isosorbide in a ratio of about 8:2 (weight ratio), and also contained a small amount of a diester derivative from sorbitan. Regarding the resulting component, 1 The measurement results of H-NMR are shown below.

[0546] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 1.00-1.80 (m, 19H), 1.90-2.20 (m, 5H), 3.6-4.9 (m, 6.4H), 5.1-5.3 (m, 0.6H), 5.71 (brs, 1H)

[0547] Mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl)sorbitan is also referred to as C20 sorbitan ester.

[0548] The resulting component was used as a mono-O-(3,7,11,15-tetramethylhexadec-2-enoyl)sorbitan component (C20 sorbitan ester component) in the examples described later.

[0549] (2) Synthesis of mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol

[0550] [Chem. 29]

[0551]

[0552] To a solution of 2.50 g (5.48 mmol) of the C20 sorbitol ester component in ethyl acetate (7.5 mL) under a nitrogen atmosphere was added 0.30 g of 5% palladium on carbon. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred under an ordinary pressure hydrogen gas atmosphere at room temperature for 1 day. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium on carbon was filtered off. The filtrate was purified by silica gel column chromatography (mobile phase: ethyl acetate) to thereby obtain 2.49 g (yield 99%) of a component containing the title compound as a colorless transparent liquid. With respect to the obtained component, 1 The measurement results of H-NMR are shown below.

[0553] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.8-0.9 (m, 12H), 0.9-1.0 (m, 3H), 1.0-2.0 (m, 22H), 2.1-2.4 (m, 2H), 3.5-4.9 (m, 6.4H), 5.0-5.2 (m, 0.6H)

[0554] Mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol is also called saturated C20 sorbitol ester.

[0555] The obtained component was used as a mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol component (saturated C20 sorbitol ester component) in the Examples described later.

[0556] (3) Synthesis of mono O-(3,7,11,15-tetramethylhexadecanoyl) sorbitol

[0557] [Chem. 30]

[0558]

[0559] To a solution of 7.5 g (99 mmol) of propylene glycol and 60 mg (0.44 mmol) of potassium carbonate in dry N,N-dimethylformamide (22 mL) was added 10.0 g (30.8 mmol) of 3,7,11,15-tetramethylhexadec-2- enoic acid methyl ester slowly dropwise at 85°C under reduced pressure of 60 to 70 mmHg and a stream of nitrogen, and stirred for 3 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 50 mL), washed with water, saturated sodium hydrogen carbonate aqueous solution, saturated brine (2 times), and 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 70:30), whereby 8.18 g of the title compound was obtained as a slightly yellow transparent liquid with low viscosity (yield 72%). With respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0560] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 1.00-1.80 (m, 22H), 1.90-2.20 (m, 6H), 3.6-3.70 (m, 0.72H), 3.85-4.2 (m, 1.93H), 5.0 (m, 0.36H), 5.71 (br s, 1H)

[0561] Mono 0-(3,7,11,15-tetramethylhexadecanoyl) propylene glycol is also called C20 propylene glycol ester.

[0562] (4) Synthesis of Mono 0-(3,7,11,15-tetramethylhexadecanoyl) propylene glycol

[0563] [Chemical 31]

[0564]

[0565] To a solution of 3.00 g (8.14 mmol) of Mono 0-(3,7,11,15-tetramethylhexadecanoyl) propylene glycol in ethyl acetate (9 mL) was added 0.36 g of 5% palladium-carbon under a nitrogen atmosphere. After replacing the nitrogen gas in the system with hydrogen gas, it was stirred for 1 day under an ordinary pressure hydrogen gas atmosphere at room temperature. After replacing the hydrogen gas in the system with nitrogen gas, the 5% palladium-carbon was filtered off. The filtrate was purified by silica gel column chromatography (ethyl acetate), whereby 2.96 g of the title compound was obtained as a colorless transparent liquid with low viscosity (yield 98%). With respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0566] 1H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.8-0.9 (m, 12H), 0.9-1.0 (m, 3H), 1.0-2.0 (m, 25H), 2.1-2.4 (m, 2H), 3.55-3.70 (m, 0.72H), 3.85-4.15 (m, 1.93H), 4.98 (m, 0.36H)

[0567] Mono O-(3,7,11,15-tetramethylhexadecanoyl) glycerol is also called saturated C20 glycerol ester.

[0568] (5) Synthesis of mono O-(4,8,12,16-tetramethylheptadecan-3-enoyl) sorbitol

[0569] [Chemical Formula 32]

[0570]

[0571] After 3.39 g (10 mmol) of 4,8,12,16-tetramethylheptadecan-3- enoic acid methyl ester and 2.74 g (15.0 mmol) of 90 wt% aqueous sorbitol (Sorbitol M-90, Sanoh Chemical Industry Co., Ltd., containing 90% solid content and 10% moisture, content in solid content: sorbitol 79% to 84%, isosorbide 15% to 18%) were added to a reaction vessel at room temperature, stirring was performed at 120°C and 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 0.11 g (2.0 mmol) of sodium methoxide and 4 mg of sodium hypophosphite monohydrate were added, and stirring was performed at 160°C and 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 54 mg (1.0 mmol) of sodium methoxide was added, and stirring was performed at 160°C and 8 kPa for 1 hour. After the pressure reduction was released with nitrogen, 54 mg (1.0 mmol) of sodium methoxide was further added, and stirring was performed at 160°C and 8 kPa for 1.5 hours. After cooling to 60°C, 10 mL of ethyl acetate and 10 mL of 0.5M hydrochloric acid were added while stirring. To the resulting reaction liquid, 50 mL of ethyl acetate was added, and extraction was performed. The extracted liquid was washed with saturated sodium bicarbonate water, saturated brine, dried with magnesium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography (mobile phase: ethyl acetate / hexane mixture) to obtain 1.79 g (yield 38%) of a component containing the title compound as a light brown transparent liquid. The resulting component contained mono O-(4,8,12,16-tetramethylheptadecan-3-enoyl) sorbitol and mono O-(4,8,12,16-tetramethylheptadecan-3-enoyl) isosorbide in a ratio of about 8:2 (weight ratio), and also contained a small amount of a diester derivative from sorbitol. With respect to the resulting component, 1 The measurement results of H-NMR are shown below.

[0572] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.90 (m, 12H), 1.00-1.60 (m, 19H), 1.64 and 1.72 (s, 3H), 2.00 (t, J = 7.4 Hz, 2H), 3.10 (d, J = 7.2 Hz, 2H), 3.5-4.9 (m, 6.4H), 5.0-5.2 (m, 0.6H), 5.30 (t, J = 7.2 Hz, IH)

[0573] Mono O-(4,8,12,16-tetramethylheptadec-3-enoyl) sorbitol is also called C21 sorbitan ester.

[0574] The obtained component was used as a Mono O-(4,8,12,16-tetramethylheptadec-3-enoyl) sorbitol component (C21 sorbitan ester component) in the Examples described later.

[0575] (6) Synthesis of Mono O-(4,8,12,16-tetramethylheptadec-3-enoyl) propylene glycol

[0576] [Chemical Formula 33]

[0577]

[0578] To a solution of 2.4 g (32 mmol) of propylene glycol and 19 mg (0.14 mmol) of potassium carbonate in dry N,N-dimethylformamide (7 mL) was added 3.39 g (10 mmol) of 4,8,12,16-tetramethylheptadec-3-enoic acid methyl ester dropwise at 85°C under a reduced pressure of 60 to 70 mmHg and a stream of nitrogen, and stirred at the same temperature for 3 hours. The obtained reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 30 mL), washed with water, saturated sodium hydrogen carbonate aqueous solution, saturated brine (2 times), and 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 70:30), whereby 2.91 g (yield 76%) of the title compound was obtained as a slightly yellow transparent liquid having low viscosity. The obtained compound was identified by1H-NMR. 1 The measurement results of1H-NMR are shown below.

[0579] 1H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.90 (m, 12H), 1.00-1.60 (m, 22H), 1.62 and 1.74 (s, 3H), 2.02 (t, J = 7.1 Hz, 2H), 3.10 (d, J = 7.0 Hz, 2H), 3.55-3.70 (m, 0.72H), 3.85-4.15 (m, 1.92H), 4.98 (m, 0.36H), 5.31 (t, J = 7.0 Hz, IH)

[0580] Mono 0-(4,8,12,16-tetramethylheptadec-3-enoyl)glycerol is also called C21 glycerol ester.

[0581] (7) Synthesis of Mono 0-(5,9,13,17-tetramethyloctadec-4-enoyl)ethanediol

[0582] [Chem. 34]

[0583]

[0584] To a solution of 2.0 g (32 mmol) of ethylene glycol and 19 mg (0.14 mmol) of potassium carbonate in dry N,N-dimethylformamide (7 mL) was added 3.53 g (10.0 mmol) of 5,9,13,17-tetramethyloctadec-4-enoic acid methyl ester slowly dropwise at 85°C under reduced pressure of 60-70 mmHg and a stream of nitrogen, and stirred for 3 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 30 mL), washed with water, saturated sodium bicarbonate water, and saturated brine (2 times), and dried over magnesium sulfate. The residue obtained by filtration and concentration was purified by silica gel column chromatography (mobile phase: hexane / ethyl acetate = 100:0-70:30), whereby 2.85 g (yield 74%) of the title compound was obtained as a slightly yellow transparent liquid with low viscosity. With respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0585] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 0.95-1.70 (m, 22H), 1.96 (td, J = 7.5, 18 Hz, 2H), 2.25-2.4 (m, 4H), 3.68 (t, J = 6.4 Hz, 2H), 4.10 (t, J = 6.4 Hz, 2H), 5.08 (m, IH)

[0586] Mono 0-(5,9,13,17-tetramethyloctadec-4-enoyl)ethanediol is also called C22 glycerol ester.

[0587] (8) Synthesis of mono O-(5, 9, 13, 17-tetramethyloctadec-4-enoyl) 1, 3-butanediol

[0588] [Chem. 35]

[0589]

[0590] To a solution of 2.9 g (32 mmol) of 1, 3-butanediol and 19 mg (0.14 mmol) of potassium carbonate in dry N, N-dimethylformamide (7 mL) was added 3.53 g (10.0 mmol) of methyl 5, 9, 13, 17-tetramethyloctadec-4-enoate slowly dropwise at 85°C under reduced pressure of 60 to 70 mmHg and a stream of nitrogen, and stirred for 3 hours at the same temperature. The resulting reaction solution was diluted with a mixed solvent of ethyl acetate / hexane (1:1, 30 mL), washed with water, saturated sodium bicarbonate water, and saturated brine (2 times), and 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 70:30), whereby 3.02 g of the title compound was obtained as a slightly yellow transparent liquid with low viscosity (yield 73%). With respect to the obtained compound, 1 The measurement results of H-NMR are shown below.

[0591] 1 H-NMR spectrum (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 0.95-1.80 (m, 25H), 1.96 (td, J = 7.6, 18.6 Hz, 2H), 2.25-2.35 (m, 4H), 3.5-3.7 (m, 0.5H), 3.86 (m, 0.75H), 4.12 (m, 0.75H), 4.35 (ddd, J = 5.4, 8.3, 13.7 Hz, 0.75H), 5.08 (m, 1.25H)

[0592] Mono O-(5, 9, 13, 17-tetramethyloctadec-4-enoyl) 1, 3-butanediol is also called C22 butanediol ester.

[0593] (9) Synthesis of mono O-(5, 9, 13, 17-tetramethyloctadec-4-enoyl) 3-methyl-1, 3-butanediol

[0594] [Chem. 36]

[0595]

[0596] To a solution of 3.3 g (32 mmol) of 3-methyl-l,3-butanediol and 19 mg (0.14 mmol) of potassium carbonate in dry N,N-dimethylformamide (7 mL) was slowly added 3.53 g (10.0 mmol) of 5,9,13,17-tetramethyloctadeca-4- enoic acid methyl ester at 85°C under reduced pressure of 60 to 70 mmHg and a nitrogen stream, and stirred for 3 hours at the same temperature. The resulting reaction solution was diluted with ethyl acetate / hexane mixed solvent (1:1, 30 mL), washed with water, saturated sodium hydrogen carbonate aqueous solution, saturated brine (2 times), and 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 70:30), whereby 2.94 g (yield 69%) of the title compound was obtained as a slightly yellow transparent liquid with low viscosity. The obtained compound, 1 The measurement results of H-NMR are shown below.

[0597] 1 H-NMR chart (300 MHz, CDC13, TMS) δ: 0.80-0.95 (m, 12H), 0.95-1.70 (m, 28H), 1.84 (t, J = 6.9 Hz, 2H), 1.96 (td, J = 7.7, 18.3 Hz, 2H), 2.25-2.35 (m, 4H), 4.25 (t, J = 6.9 Hz, 2H), 5.08 (m, IH)

[0598] Mono-O-(5,9,13,17-tetramethyloctadeca-4-enoyl) 3-methyl-l,3-butanediol is also called C22 isoprenoid ester.

[0599] [Example 18] Preparation of precursor formulations, gel formation test, and analysis of liquid crystal structure (3)

[0600] The isoprenoid-type lipids, the amphiphilic compounds synthesized in Example 17, SPC (LIPOID S100, Lipoid Co.) as a phospholipid, and alcohol were mixed in the mixing ratio shown in Table 13 below. The resulting mixture was dissolved in a water bath at 40°C or lower, whereby the precursor formulations Nos. 176 to 184 shown in Table 13 were prepared.

[0601] Note that C20 propanediol ester, saturated C20 propanediol ester, C21 propanediol ester, C22 ethylene glycol ester, C22 butanediol ester, and C22 isoprenoid ester do not form a gel when mixed with water alone, and thus are not self-organizing lipids (SOL).

[0602] For the precursor preparations No. 176 to 184, a gel formation test was performed. To a vial, an excess of water for injection (0.5 to 2 mL or so) was added with a portion of each of the precursor preparations (100 to 300 mg or so), and mixing operation was performed at room temperature (25°C) using a medicine spoon and / or a vortex mixer. As a result, in all of the precursor preparations No. 176 to 184, a gel-like composition which was separated in the excess of water for injection (aqueous medium) and which was colorless transparent to white turbid in appearance was obtained.

[0603] The gel-like compositions obtained from the precursor preparations No. 182 to 184 would exhibit the HII liquid crystal phase as with the preparations No. 146 and 147.

[0604] [Table 13]

[0605]

[0606] All publications, patents, and patent applications cited in this specification are herein incorporated by reference.

Claims

1. A non-laminar liquid crystal forming composition comprising an amphiphilic compound represented by the following general formula (I) and a phospholipid, [Chemistry 1] In the formula, X and Y together represent oxygen atoms, and n represents 2. Indicates a single bond or a double bond. R represents a hydrophilic group formed by removing one hydroxyl group from any of the following groups: glycerol, erythritol, pentaerythritol, diglycerol, sorbitol, isosorbide, propylene glycol, ethylene glycol, butylene glycol, and isopentyl glycol; m represents 2. The weight ratio of the amphiphilic compound to the phospholipid is 70:30 to 30:

70.

2. The composition according to claim 1, wherein, The amphiphilic compounds are represented by the following general formulas (II), (III), or (IV): [Chemistry 2] In the formula, X and Y together represent oxygen atoms, and n represents 2. R represents the hydrophilic group obtained by removing one hydroxyl group from any of the following groups: glycerol, erythritol, pentaerythritol, diglycerol, sorbitol, isosorbide, propylene glycol, ethylene glycol, butylene glycol, and isopentyl glycol; m represents 2.

3. The composition according to claim 1 or 2, wherein, The weight ratio of the amphiphilic compound to the phospholipid is 50:50 to 30:

70.

4. The composition according to claim 1 or 2, wherein, The weight ratio of the amphiphilic compound to the phospholipid is 45:55 to 30:

70.

5. The composition according to any one of claims 1 to 4, wherein, Phospholipids are selected from the group consisting of phosphatidylcholine and phosphatidylethanolamine.

6. The composition according to any one of claims 1 to 5, wherein, Phospholipids are selected from the group consisting of: soybean phosphatidylcholine, egg yolk phosphatidylcholine, myristoyl phosphatidylcholine, dioleoyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine.

7. The composition according to any one of claims 1 to 6, wherein, In the general formula (I), R represents a hydrophilic group obtained by removing one hydroxyl group from any of the following groups: erythritol, pentaerythritol, diglycerides, sorbitol, isosorbide, propylene glycol, ethylene glycol, butylene glycol, and isopentyl glycol, and m represents 2.

8. The composition according to any one of claims 1 to 7, wherein, In the general formula (I), R represents a hydrophilic group obtained by removing one hydroxyl group from any of the following groups: sorbitol, isosorbide, propylene glycol, ethylene glycol, butanediol, and isopentyl glycol, and m represents 2.

9. The composition according to any one of claims 1 to 8, wherein, In the general formula (I), R represents the hydrophilic group after removing one hydroxyl group from sorbitol or isosorbide, and m represents 2.

10. The composition according to claim 1 or 2, wherein, The amphiphilic compound is selected from the group consisting of the following compounds: Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)glycerol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)glycerol; Mono-O-(5,9,13,17-tetramethyloctadec-4,8,12,16-tetraenoyl)glycerol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)erythritol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)pentaerythritol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)pentaerythritol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)diglycerol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) sorbitol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)-dehydrated sorbitol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)isosorbitol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)isosorbitol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)propanediol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)propanediol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)ethylene glycol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)1,3-butanediol; and Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)3-methyl-1,3-butanediol.

11. A non-layered liquid crystal forming composition comprising an amphiphilic compound represented by the following general formula (I) and a phospholipid, [Chemistry 1] In the formula, X and Y together represent oxygen atoms, and n represents 2. Indicates a single bond or a double bond. R represents the hydrophilic group obtained by removing one hydroxyl group from any of the following groups: sorbitol, isosorbide, propylene glycol, ethylene glycol, butanediol, and isopentyl glycol; m represents 1. The weight ratio of the amphiphilic compound to the phospholipid is 50:50 to 30:

70.

12. The composition according to claim 11, wherein, The amphiphilic compounds are represented by the following general formulas (II), (III), or (IV): [Chemistry 2] In the formula, X and Y together represent oxygen atoms, and n represents 2. R represents the hydrophilic group obtained by removing one hydroxyl group from any of the following groups: sorbitol, isosorbide, propylene glycol, ethylene glycol, butanediol, and isopentyl glycol, and m represents 1.

13. The composition according to claim 11 or 12, wherein, The weight ratio of the amphiphilic compound to the phospholipid is 40:60 to 30:

70.

14. The composition according to claim 11 or 12, wherein, The weight ratio of the amphiphilic compound to the phospholipid is 45:55 to 30:

70.

15. The composition according to any one of claims 11-14, wherein, Phospholipids are selected from the group consisting of phosphatidylcholine and phosphatidylethanolamine.

16. The composition according to any one of claims 11-15, wherein, Phospholipids are selected from the group consisting of: soybean phosphatidylcholine, egg yolk phosphatidylcholine, myristoyl phosphatidylcholine, dioleoyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine.

17. The composition according to any one of claims 11-16, wherein, In the general formula (I), R represents a hydrophilic group obtained by removing one hydroxyl group from any of the following groups: sorbitol, isosorbide, ethylene glycol, butanediol, and isopentyl glycol, and m represents 1.

18. The composition according to any one of claims 11-17, wherein, In the general formula (I), R represents the hydrophilic group after removing one hydroxyl group from sorbitol or isosorbide, and m represents 1.

19. The composition according to claim 11 or 12, wherein, The amphiphilic compound is selected from the group consisting of the following compounds: Mono-O-(5,9,13-trimethyltetradec-4-enoyl) dehydrated sorbitol; Mono-O-(5,9,13-trimethyltetradecanoyl)-dehydrated sorbitol; and Mono-O-(5,9,13-trimethyltetradec-4-enoyl)propanediol.

20. The composition according to any one of claims 1 to 19, wherein it contains at least one of an oil and an organic solvent.

21. The composition according to any one of claims 1 to 20, wherein, The composition is a non-layered liquid crystal composition that also contains an aqueous medium.

22. The composition according to claim 21, wherein, The composition is a non-layered liquid crystal emulsion composition that also contains a surfactant.

23. The composition according to any one of claims 1 to 20, wherein, The composition is a liquid crystal precursor composition capable of forming a non-laminar liquid crystal in the presence of an aqueous medium.

24. A pharmaceutical preparation comprising the composition according to any one of claims 1 to 23.

25. The pharmaceutical preparation according to claim 24, wherein, The composition also contains a drug, and the drug formulation is a sustained-release formulation.

26. The pharmaceutical preparation according to claim 25, wherein, The drug is a gonadotropin-releasing hormone (GnRH) agonist.

27. The pharmaceutical preparation according to claim 26, wherein, The GnRH agonist is leuprolide or its salt.

28. The pharmaceutical preparation according to any one of claims 24-27, wherein it is a spray, an injection, or a long-acting preparation.

29. The pharmaceutical preparation according to any one of claims 24-27, wherein it is an aerosol.

30. Use of the composition according to any one of claims 1 to 23 in the preparation of a pharmaceutical formulation for preventing adhesion of biological tissues.

31. The use according to claim 30, wherein the pharmaceutical preparation is a spray, an injection, or a long-acting preparation.

32. The use according to claim 30, wherein the pharmaceutical preparation is an aerosol.

33. An amphiphilic compound or a salt thereof represented by the following general formula (I'): [Chemistry 3] In the formula, X and Y together represent oxygen atoms, n represents 2, and m represents 1 or 2. It indicates a single or double bond, and R represents the hydrophilic group after removing one hydroxyl group from sorbitol or isosorbide.

34. The compound or a salt thereof according to claim 33, wherein, The amphiphilic compound is selected from the group consisting of the following compounds: Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl) sorbitol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)-dehydrated sorbitol; Mono-O-(5,9,13-trimethyltetradec-4-enoyl) dehydrated sorbitol; Mono-O-(5,9,13-trimethyltetradecanoyl) dehydrated sorbitol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)isosorbitol; and Mono-O-(5,9,13,17-tetramethyloctadecanoyl)isosorbitol.

35. An amphiphilic compound or a salt thereof represented by the following general formula (III) or (IV): In the formula, X and Y together represent oxygen atoms, n represents 2, m represents 1 or 2, and R represents the hydrophilic group after removing one hydroxyl group from propylene glycol, ethylene glycol, butanediol, or isopentyl glycol.

36. The compound or a salt thereof according to claim 35, wherein, The amphiphilic compound is selected from the group consisting of the following compounds: Mono-O-(5,9,13-trimethyltetradec-4-enoyl)propanediol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)propanediol; Mono-O-(5,9,13,17-tetramethyloctadecanoyl)propanediol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)ethylene glycol; Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)1,3-butanediol; and Mono-O-(5,9,13,17-tetramethyloctadec-4-enoyl)3-methyl-1,3-butanediol.

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