Composition containing high-purity hydrophobic ion pair

Abstract

The present disclosure provides a solution containing a high-purity ion pair [EDMPC]+[anionic surfactant]-. The solution containing a high-purity ion pair [EDMPC]+[anionic surfactant]- can be obtained by introducing a salt of [EDMPC]+ into an anion exchange resin that has adsorbed the [anionic surfactant]-.

Description

Composition containing high-purity hydrophobic ion pairs

[0001] This disclosure relates to a composition containing a high-purity hydrophobic ion pair and a method for preparing the same.

[0002] Compositions containing water-soluble ion pairs have been synthesized by methods such as ion exchange (Non-Patent Document 1) and neutralization (Non-Patent Document 2), and have been prepared by replacing ions with other desired ions through ion exchange reactions.

[0003] A composition containing hydrophobic ion pairs is disclosed, for example, in Patent Document 1. In Patent Document 1, after the ion pair has been formed, the desired ion pair is extracted by adding anions to the system and phase separation. However, this method results in limited yield and low purity.

[0004] WO2021 / 192861A

[0005] AK Burrell et al., Green Chem., 9, 449 (2007)K. Ohira et al, Chem. Sus. Chem., 5, 388 (2012)

[0006] This disclosure provides a composition containing a high-purity hydrophobic ion pair and a method for preparing the same. The inventors were able to obtain an ion pair of a hydrophobic cation and a hydrophilic anion with extremely high purity by reacting an anion exchange resin on which a hydrophobic anion has been adsorbed in the presence of an organic solvent with an ion pair of a hydrophobic cation and a hydrophilic anion, and then extracting the mixture with an organic solvent.

[0007] The present invention may provide the following inventions: (1) A composition comprising a hydrophobic cation and a hydrophobic anion, wherein the hydrophobic cation and the hydrophobic anion together constitute 80% by weight or more (preferably 90% by weight or more) of the total weight of hydrophobic molecules in the composition. (2) The composition according to (1), wherein the hydrophobic cation comprises a diglycerol group containing a higher fatty acid ester, and / or the hydrophobic anion is an anionic surfactant, preferably an anionic surfactant containing a medium-chain or long-chain carbon chain such as a higher fatty acid or a higher alkyl sulfate. (3) The composition according to (1) or (2), wherein the hydrophobic cation comprises 1,2-dimyristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC). (4) The composition according to any one of (1) to (3), wherein the hydrophobic anion comprises a higher fatty acid such as linoleic acid, or a higher alkyl sulfate such as dodecyl sulfate. (5) The composition according to any one of (1) to (4) above, further comprising a solvent, wherein the solvent comprises an alcohol. (6) The composition according to (5) above, wherein the alcohol comprises a monohydric lower alcohol. (7) The composition according to (5) above, wherein the alcohol comprises a polyhydric alcohol. (8) The composition according to any one of (1) to (7) above, wherein the hydrophobic cation comprises 1,2-dimiristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC), the hydrophobic anion comprises an anionic surfactant, preferably an anionic surfactant comprising a medium-chain or long-chain carbon chain such as a higher fatty acid or a higher alkyl sulfate, the solvent comprises an alcohol, and after storage at -20°C for 2 months, the total amount of EDMPC and the hydrophobic cation is 80% by weight or more (preferably 90% by weight or more) of the total hydrophobic molecules in the composition. (9) The composition according to (8) above, which does not contain dimyristoylphosphatidylcholine (DMPC), or contains DMPC, but the DMPC content is less than 5% by weight. (10) The composition according to (9) above, wherein the DMPC content is 10% by weight or less (preferably 5% by weight or less) after storage at -20°C for two months.

[0008] (21) A composition comprising EDMPC and a hydrophobic anion (preferably a higher fatty acid such as linoleic acid), wherein the composition is electrically neutralized and the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more (preferably 90% by weight or more) of the total weight of hydrophobic molecules in the composition. (22) A composition comprising EDMPC and a hydrophobic anion (preferably a higher fatty acid such as linoleic acid), wherein the composition is electrically neutralized and the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more (preferably 90% by weight or more) of the total weight of molecules in the composition. (23) The composition according to (21) or (22) above, further comprising an alcohol. (24) The composition according to (23) above, wherein the alcohol comprises ethanol. (25) The composition according to (23) above, wherein the alcohol comprises a polyol. (26) The composition according to (25) above, wherein the polyol comprises a polyalkylene glycol. (27) The composition according to any of the above, wherein after storage at -20°C for two months, the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more (preferably 90% by weight or more) of the total weight of hydrophobic molecules in the composition. (28) The composition according to any of the above, wherein after storage at -20°C for two months, the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more of the total weight of molecules in the composition. (29) The composition according to (27) above, wherein the DMPC content is 10% by weight or less (preferably 5% by weight or less) after storage at -20°C for two months. (30) The composition according to (28) above, wherein the DMPC content is 10% by weight or less (preferably 5% by weight or less) after storage at -20°C for two months. (31) The composition according to any of the above, which has been stored at -20°C for one month or more (for example, two months or more). (32) The composition according to any of the above, which is stored at -20°C for one month or more (for example, two months or more) and has a DMPC content of 10% by weight or less (preferably 5% by weight or less).

[0009] (41) A composition comprising EDMPC and a hydrophobic anion (preferably a higher alkyl sulfate such as dodecyl sulfate), wherein the composition is electrically neutralized and the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more (preferably 90% by weight or more) of the total weight of hydrophobic molecules in the composition. (42) A composition comprising EDMPC and a hydrophobic anion (preferably a higher alkyl sulfate such as dodecyl sulfate), wherein the composition is electrically neutralized and the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more (preferably 90% by weight or more) of the total weight of molecules in the composition. (43) The composition according to (41) or (42) above, further comprising an alcohol. (44) The composition according to (43) above, wherein the alcohol comprises ethanol. (45) The composition according to (43) above, wherein the alcohol comprises a polyol. (46) The composition according to (45) above, wherein the polyol comprises a polyalkylene glycol. (47) The composition according to any one of the above, wherein after storage at -20°C for two months, the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more (preferably 90% by weight or more) of the total weight of hydrophobic molecules in the composition. (48) The composition according to any one of the above, wherein after storage at -20°C for two months, the total amount of the hydrophobic cation and the hydrophobic anion is 80% by weight or more of the total weight of molecules in the composition. (49) The composition according to (47) above, wherein the DMPC content is 10% by weight or less (preferably 5% by weight or less) after storage at -20°C for two months. (50) The composition according to (48) above, wherein the DMPC content is 10% by weight or less (preferably 5% by weight or less) after storage at -20°C for two months. (51) The composition according to any one of the above, which has been stored at -20°C for one month or more (for example, two months or more). (52) The composition according to any of the above, which is stored at -20°C for one month or more (for example, two months or more) and has a DMPC content of 10% by weight or less (preferably 5% by weight or less).

[0010] [EDMPC] before ion exchange + [TfO] - and [EDMPC] after ion exchange + [Linoleic acid] - of 1 H-NMR, 19 F-NMR, and31 Shows the spectrum of P-NMR. [EDMPC] after ion exchange + [Linoleic acid] - Shows the absorption spectra by infrared absorption spectroscopy (IR) of various molecules containing [EDMPC]. + [Linoleic acid] - Shows the spectrum of 31P-NMR indicating the storage instability during drying under reduced pressure. [EDMPC] + [Linoleic acid] - Shows the chromatogram by high performance liquid chromatography (HPLC) after drying under reduced pressure of [EDMPC]. + [Linoleic acid] - Shows the chromatogram by HPLC of the dried product under reduced pressure of [EDMPC] and an ethanol solution. [EDMPC] in the ethanol solution + [Linoleic acid] - Shows the storage stability (-20 °C condition). [EDMPC] + [Linoleic acid] - Shows the chromatogram by HPLC of polyhydric alcohol solutions such as propylene glycol (PG) and butylene glycol (BG) of [EDMPC]. [EDMPC] before ion exchange + [TfO] - and [EDMPC] after ion exchange + [DS] - of １ H-NMR, １９ F-NMR, and ３１ P-NMR spectra. [EDMPC] + [DS] - Shows the MALDI-HRMS spectrum. [EDMPC] stored at -30 °C for 8 months + [DS] - of １ H-NMR and ３１ P-NMR spectra. Immediately after preparation and after 6 months of storage of [EDMPC] + [DS] - Shows the HPLC chromatogram. [EDMPC] + [DS] - Shows the storage stability of EDMPC in an ethanol solution containing [DS].

[0011] <Definitions of Terms> In this specification, the term "contains" means that the invention may contain further third components not listed. In this specification, the term "consisting of" means that the invention does not contain third components not listed, or contains third components in amounts that are unavoidable in the manufacture or below the detection limit. In this specification, the term "substantially consisting of" means that the invention may contain third components that do not affect the effect of the invention.

[0012] In this specification, "hydrophobic cation" refers to a hydrophobic molecule having a cationic group, which exhibits cationic properties as a whole. The hydrophobic portion of a hydrophobic cation may contain fatty acid chains (e.g., saturated or unsaturated fatty acid chains), and may include fatty acid esters. In this specification, "hydrophobic anion" refers to a hydrophobic molecule having anionic group, which exhibits anionic properties as a whole. The hydrophobic portion of a hydrophobic anion may contain fatty acid chains, and may include fatty acid esters. Hydrophobic cations and hydrophobic anions may each have surfactant properties. Hydrophobic cations and hydrophobic anions may be cationic surfactants and anionic surfactants, respectively. The HLB values ​​of hydrophobic cations and hydrophobic anions may be, for example, 6 or less, preferably 5 or less, more preferably 4 or less (e.g., 2-5, 2-3, 2-4, 3-4, 3-5, or 4-5).

[0013] In this specification, "HLB value" is a numerical value representing the balance between hydrophilicity and hydrophobicity of a surfactant. Nonionic substances that do not have hydrophilic groups have an HLB value of 0, and nonionic substances that only have hydrophilic groups have an HLB value of 20. A smaller HLB value indicates greater hydrophobicity, and a higher HLB value indicates greater hydrophilicity. The HLB value of a nonionic substance is calculated as (sum of molecular weights of hydrophilic parts / molecular weight) × 20. Surfactants with an HLB value of 3 to 6 can partially disperse in water and function as emulsifiers. The HLB value of an ionic substance is estimated using the Davies formula (HLB = 7 + ΣH_i - 0.475 × n), where n is the number of lipophilic carbons and ΣH_i is the sum of the group constants of the hydrophilic groups. For ionic substances, the HLB value can exceed 20 and reach a maximum of approximately 40. Because apparent hydrophilicity varies depending on acidic and saltic forms, pH, counterion species, and degree of dissociation, unless otherwise specified, the dissociation state in an aqueous solution at 25°C and near neutral pH is assumed. Therefore, the functional guidelines for nonionic surfactants (e.g., emulsification at HLB 3-6) are not directly applicable, and for ionic surfactants, they are treated as reference indicators for application selection.

[0014] In this specification, “fatty acid chain” refers to saturated or unsaturated straight or branched hydrocarbon chains such as straight or branched alkyls, straight or branched alkenyls, and straight or branched alkynyls. In this specification, “fatty acid” refers to a fatty acid chain having a carboxyl group at its terminus. Short-chain fatty acids are fatty acid chains having 4 to 6 carbon atoms, medium-chain fatty acids are fatty acid chains having 7 to 13 carbon atoms, and long-chain fatty acids are fatty acid chains having 14 or more carbon atoms (e.g., 14 to 22). However, the carbon number of fatty acids includes the carbon atoms of the carboxyl group. Higher fatty acids mean fatty acid chains and fatty acids having 6 or more carbon atoms. Fatty acid chains and fatty acids may contain one or more double or triple bonds.

[0015] Alkyl groups include linear or branched alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, secondary butyl (sec-butyl), and tert-butyl (tert-butyl) groups. While not particularly limited, alkyl groups include C1 alkyl, C2 alkyl, and C1-3 Alkyl, C 1-4 Alkyl, C 1-5 Alkyl and C 1-6 Examples include lower alkyl groups such as alkyl groups, and higher alkyl groups with five or more carbon atoms. An alkenyl group is a compound in which one C-C bond in an alkyl group is replaced by a double bond, and an alkynyl group is a compound in which one C-C bond in an alkyl group is replaced by a triple bond.

[0016] In this specification, "solvent" means a liquid that dissolves the solute. When the solute is a hydrophobic substance, the solvent may be a lipid-soluble solvent such as an alcohol.

[0017] In this specification, "alcohol" means a hydrocarbon having one or more hydroxyl groups. An alcohol may, for example, be a fatty chain having one or more hydroxyl groups. "n-valent alcohol" means having n hydroxyl groups. A polyhydric alcohol is an alcohol having a large number (e.g., 10 or more) hydroxyl groups.

[0018] <Compositions of the Disclosure> The Disclosure provides compositions comprising a hydrophobic cation, a hydrophobic anion, and a solvent. In one preferred embodiment, the hydrophobic cation and the hydrophobic anion each have an HLB value of 5 or less. In one preferred embodiment, the hydrophobic cation and the hydrophobic anion are electrically neutralized in the composition.

[0019] Electrical neutralization can be achieved by the presence of stoichiometrically equal amounts of the cationic group of the hydrophobic cation and the anionic group of the hydrophobic anion.

[0020] In one preferred embodiment, both the hydrophobic cation and the hydrophobic anion are monovalent, having one cation and one anion, respectively. Monovalent means that the hydrophobic cation has one cationic group. Monovalent means that the hydrophobic anion has one anionic group. In one embodiment, the hydrophobic cation and the hydrophobic anion are independently monovalent or polyvalent.

[0021] In a preferred embodiment, the composition may further contain a solvent. Examples of solvents include volatile solvents such as cyclohexane and alcohols.

[0022] In one preferred embodiment, the hydrophobic cations and hydrophobic anions in the composition may each be 50% or more, 60% or more, 70% or more, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more, based on the dry weight of the total cations and anions contained in the composition.

[0023] In one preferred embodiment, the hydrophobic cations and hydrophobic anions in the composition may be 50% or more by weight, 60% or more by weight, 70% or more by weight, 80% or more by weight, 85% or more by weight, 90% or more by weight, 95% or more by weight, 96% or more by weight, 97% or more by weight, 98% or more by weight, or 99% or more by weight, based on the dry weight of the total hydrophobic cations and total hydrophobic anions contained in the composition.

[0024] In one preferred embodiment, the total amount of hydrophobic cations and hydrophobic anions in the composition may be 50% or more, 60% or more, 70% or more, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more by weight of the dry weight of all hydrophobic molecules contained in the composition.

[0025] In one preferred embodiment, the hydrophobic cation and hydrophobic anion in the composition each have a cationic group and an anionic group, and the hydrophobic portion contains a fatty chain, and the total amount of the hydrophobic cation and hydrophobic anion may be 50% or more by weight, 60% or more by weight, 70% or more by weight, 80% or more by weight, 85% or more by weight, 90% or more by weight, 95% or more by weight, 96% or more by weight, 97% or more by weight, 98% or more by weight, or 99% or more by weight, based on the dry weight of the total molecule including the fatty chain.

[0026] In a preferred embodiment, the composition may further contain a pharmaceutically active ingredient (API).

[0027] In one preferred embodiment, the composition further comprises a pharmaceutically active ingredient (API) dispersed in the composition, which together with a surfactant comprising hydrophobic cations and hydrophobic anions in the composition, forming solid particles, the total amount of hydrophobic cations and hydrophobic anions in the solid particles may be 50% or more by weight, 60% or more by weight, 70% or more by weight, 80% or more by weight, 85% or more by weight, 90% or more by weight, 95% or more by weight, 96% or more by weight, 97% or more by weight, 98% or more by weight, or 99% or more by weight, based on the dry weight of the surfactant in the solid particles. Compositions containing solid particles and in which the solid particles are dispersed are disclosed, for example, in WO2009 / 066457A and WO2021 / 192861A.

[0028] In one preferred embodiment, the composition further comprises a pharmaceutically active ingredient (API) dispersed in the composition, which forms a complex with an aqueous composition containing hydrophobic cations and hydrophobic anions, and the total amount of hydrophobic cations and hydrophobic anions in the solid particles may be 50% or more by weight, 60% or more by weight, 70% or more by weight, 80% or more by weight, 85% or more by weight, 90% or more by weight, 95% or more by weight, 96% or more by weight, 97% or more by weight, 98% or more by weight, or 99% or more by weight, based on the dry weight of the surfactant in the complex. Such compositions are disclosed, for example, in WO2023 / 234250A.

[0029] In one preferred embodiment, the hydrophobic cation may be a cationic lipid. In one preferred embodiment, the hydrophobic anion may be an anionic lipid. In a more preferred embodiment, the hydrophobic cation may be a cationic lipid, and the hydrophobic anion may be an anionic lipid. In one preferred embodiment, the lipid includes a fatty acid chain.

[0030] In some preferred embodiments, the hydrophobic cation may comprise a higher fatty chain and a cationic group. In some preferred embodiments, the hydrophobic cation may comprise a long fatty chain and a cationic group. In some preferred embodiments, the hydrophobic cation comprises a diglycerol group containing two long fatty chains and a cationic group. In some preferred embodiments, the cationic group may comprise a quaternary ammonium cation. In some preferred embodiments, the cationic groups are each independently -CH２ -N-(R １ ) (Caution ２ ) (Caution ３ ) ＋ Base {Here R １ , R ２ , and R ３ Each of these may independently be a lower alkyl group, preferably an ethyl group, or more preferably a methyl group. In one preferred embodiment, the cationic group is -CH ２ -N-(CH ３ ) ３ Is it positive or negative? ２ -N-(CH ３ ) ３ ＋ It has the following characteristics. In one preferred embodiment, the cationic group is choline. In one preferred embodiment, the cationic group is phosphatidylcholine. In one preferred embodiment, the hydrophobic cation may be 1,2-long-chain fatty acid-sn-glycero-3-ethyl-phosphatidylcholine, and more preferably 1,2-dimyristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC).

[0031] In a preferred embodiment, the hydrophobic anion may comprise a higher fatty acid chain and an anionic group. In a preferred embodiment, the hydrophobic anion may comprise a fatty acid chain (e.g., a medium-chain fatty acid chain or a long-chain fatty acid chain) and an anionic group. In a preferred embodiment, the anionic group may be a carboxyl group, a sulfonic acid group, a phosphate ester group, a phosphonic acid group, or a sulfate ester group. In a preferred embodiment, the hydrophobic anion may be a medium-chain or long-chain fatty acid. In a preferred embodiment, the hydrophobic anion may be a fatty acid having 10 to 22 carbon atoms, for example, a fatty acid having 14 to 20 carbon atoms, for example, a fatty acid having 16 to 18 carbon atoms. In a preferred embodiment, the fatty acid may be a saturated fatty acid, for example, a saturated fatty acid selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, and behenic acid. In one preferred embodiment, the fatty acid may be one or more monounsaturated fatty acids selected from the group consisting of, for example, palmitoleic acid, oleic acid, elaidic acid, and erucic acid. In one preferred embodiment, the fatty acid may be one or more polyunsaturated fatty acids selected from the group consisting of, for example, linoleic acid, α-linolenic acid, γ-linolenic acid, arachidonic acid, all-cis-5,8,11,14-eicosatetraenoic acid (EPA), and all-cis-4,7,10,13,16,19-docosahexaenoic acid (DHA).

[0032] Examples of hydrophobic anions include fatty acids (higher saturated fatty acids such as capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, and arachidic acid; monounsaturated higher fatty acids such as caprioleic acid, lauroleic acid, myristoleic acid, palmitoleic acid, oleic acid, and eicosenoic acid; polyunsaturated higher fatty acids such as linoleic acid, alpha-linolenic acid, arachidonic acid, and eicosapentaenoic acid), sulfate esters (linear higher alkyl sulfates such as caprylic acid, undecyl sulfate, lauryl sulfate, tridecyl sulfate, myristyl sulfate, pentadecyl sulfate, cetyl sulfate, heptadecyl sulfate, stearyl sulfate, nonadecyl sulfate, and arachidyl sulfate), and sulfonate-type surfactants (decylbenzenesulfonic acid, Examples include higher alkylbenzenesulfonic acids such as undecylbenzenesulfonic acid, dodecylbenzenesulfonic acid, tridecylbenzenesulfonic acid, tetradecylbenzenesulfonic acid, pentadecylbenzenesulfonic acid, hexadecylbenzenesulfonic acid, heptadecylbenzenesulfonic acid, octadecylbenzenesulfonic acid, nonadecylbenzenesulfonic acid, and eicosylbenzenesulfonic acid; and higher alkylsulfonic acids such as caprylylsulfonic acid, undecylsulfonic acid, laurylsulfonic acid, tridecylsulfonic acid, myristylsulfonic acid, pentadecylsulfonic acid, cetylsulfonic acid, heptadecylsulfonic acid, stearylsulfonic acid, nonadecylsulfonic acid, and arachilsulfonic acid. Hydrophobic anions include, for example, sulfate-based surfactants and / or sulfonate-based surfactants. As described above, hydrophobic anions preferably contain carbon chains with 8 or more, 10 or more, 12 or more, or 14 or more carbon atoms, and preferably contain carbon chains with 10 to 22, 12 to 20, 12 to 18, 12 to 16, 14 to 20, or 16 to 18 carbon atoms.

[0033] In one preferred embodiment, the hydrophobic cation is EDMPC and the hydrophobic anion is linoleic acid. In another preferred embodiment, the hydrophobic cation is EDMPC and the hydrophobic anion may be dodecyl sulfate.

[0034] If the solvent is volatile (for example, if the solvent is ethanol, methanol, or cyclohexane), the composition may contain the solvent, but may also be solvent-free after drying.

[0035] In a preferred embodiment, the solvent contains or is substantially composed of an alcohol. In a preferred embodiment, the alcohol may be 50% or more by weight, 60% or more by weight, 70% or more by weight, 80% or more by weight, 85% or more by weight, 90% or more by weight, 95% or more by weight, 96% or more by weight, 97% or more by weight, 98% or more by weight, or 99% or more by weight, relative to the total solvent. In a preferred embodiment, the alcohol contains or consists of a lower alcohol. The lower alcohol is C 1-4 These are alcohols, such as methanol, ethanol, n-propyl alcohol, isopropyl alcohol, n-butyl alcohol, isobutyl alcohol, secondary butyl alcohol (sec-butyl alcohol), and tertiary butyl alcohol (tert-butyl alcohol).

[0036] In a preferred embodiment, the solvent contains or substantially consists of a polyol. A polyol has multiple hydroxyl groups in a single molecule. Examples of polyols, though not particularly limited, include sugar alcohols, polyether polyols (e.g., polypropylene glycol, polyethylene glycol, polyoxypropylene glycol, polyoxyethylene triol, polyoxypropylene triol), polyester polyols (adipate esters, phthalate esters), and natural oil-derived polyols (soybean oil polyol, castor oil polyol). In a preferred embodiment, the polyol contains or substantially consists of polypropylene glycol or polyethylene glycol.

[0037] In a preferred embodiment, the compositions of the present disclosure consist of pharmaceutically acceptable or pharmaceutically acceptable components.

[0038] <Method for producing the composition of this disclosure> Examples of synthesis methods for ionic organic molecules include ion exchange (AK Burrell, RE Del Sesto, SN Baker, TM McCleskey, GA Baker, Green Chem., 9, 449 (2007)) and neutralization (K. Ohira, Y. Abe, K. Suzuki, M. Mizuno, Y. Amano, T. Itoh, ChemSusChem, 5, 388 (2012)). In the above case, where the ionic organic molecule is water-soluble, its purification can be carried out by an ion exchange reaction in water.

[0039] According to this disclosure, a hydrophobic cation (e.g., [EDMPC]) is added to an anion exchange resin that has adsorbed hydrophobic anions in an organic solvent (e.g., methanol or ethanol). + We discovered that by passing a solution through an anion exchange resin, hydrophilic anions (e.g., chloride salts, trifluoromethanesulfonates, etc.) can be replaced with hydrophobic anions, thereby enabling the preparation of highly pure ion aggregates (ion pairs) of hydrophobic cations and hydrophobic anions. It was also revealed that the anion exchange efficiency was extremely high. This method is considered effective for preparing ion aggregates of hydrophobic cations and hydrophobic anions.

[0040] If the hydrophobic cation is EDMPC, then EDMPC is converted to DMPC in chloroform. ３ SO ３ It can be selected by reacting with Et {Et represents an ethyl group}. The resulting compound is [EDMPC]. ＋ [TfO] － {TfO represents trifluoromethanesulfonic acid}. [TfO] － of [hydrophobic anion] － Preferably hydrophobic anions (preferably [long-chain fatty acids]) － or [long-chain alkyl sulfate] － Exchange to anionic surfactant ions having higher carbon chains (preferably [linoleic acid]) － or [dodecyl sulfate] －This can be carried out in alcohol as described above. The solvent can be removed from the resulting composition, but it is not necessary. If the solvent is not removed, the hydrophobic cation-hydrophobic anion ion pair in this composition remains stable at -20°C for more than two months, and substantially no decomposition products are produced under these storage conditions.

[0041] If the composition contains a hydrophobic cation, a hydrophobic anion, and an alcohol, it may be used as is, or the solvent may be removed before use (for example, immediately before use).

[0042] In some embodiments, an equimolar mixture of a monovalent hydrophobic cation and a monovalent hydrophobic anion can be an ionic liquid.

[0043] In one preferred embodiment, the resulting compound is [EDMPC] ＋ [Long-chain fatty acids] － And especially [EDMPC] ＋ [Linoleic acid] － It can be used, for example, as disclosed in WO2023 / 234250A. In one preferred embodiment, the resulting compound is [EDMPC] ＋ [Long-chain alkyl sulfates] － and especially [EDMPC] ＋ [Dodecyl sulfate] － This is possible. However, while WO2021 / 192861A and WO2023 / 234250A did not achieve the preparation of highly purified ion pairs, this disclosure achieves the preparation of highly purified ion pairs and is considered effective in obtaining formulations containing highly purified or compositionally controlled particles.

[0044] The purity of EDMPC can be calculated, for example, based on the following equations 1 and 2: Pure [EDMPC] + [Hydrophobic anions] - (For example, [EDMPC] + [Linoleic acid] - Theoretical value of EDMPC contained in ) = (Molecular weight of EDMPC skeleton 707.01) / ([EDMPC] + [Hydrophobic anions (e.g., linoleic acid)] -Molecular weight 985.45) ... (Equation 1) Purity (wt%) = Quantitative value of EDMPC (mg / mL) / Dry weight of 1 mL of solution (mg / mL) / Theoretical value of EDMPC × 100 ... (Equation 2)

[0045] 1. Reagents and Apparatus The reagents used in the examples are listed below. Muromac XMA-413B-Cl was obtained from Muromachi Chemical Co., Ltd. N,N'-dicyclohexylcarbodiimide (DCC), 4-dimethylaminopyridine (DMAP), myristic acid, ethyl trifluoromethanesulfonate, and sodium linoleate were obtained from Tokyo Chemical Co., Ltd. Hyflo Super Cel (trademark), sodium dodecyl sulfate (for ion pair chromatography), sodium chloride (special grade), sodium bicarbonate (special grade), sodium sulfate (special grade), phosphorus oxide (special grade), methanol (super dehydrated), chloroform (super dehydrated, with amylene added), diisopropyl ether (special grade), and ethanol (99.5% special grade) were obtained from Fujifilm Wako Pure Chemical Corporation. Choline glycerophosphate was obtained from Combi-Blocks. Ethyl acetate, chloroform, and methanol for extraction and purification were obtained from Kanto Chemical Co., Ltd.

[0046] The following purification and analytical instruments were used: Purification system: 2-channel parallel purification system Purif-espoir2 (manufactured by Shoko Science Co., Ltd.) Nuclear magnetic resonance (NMR) spectrometer: JEOL ECS 400 (manufactured by JEOL Ltd.), JEOL ECA 600 (manufactured by JEOL Ltd.) Infrared spectrometer (IR): JASCO-FT / IR-4200 (manufactured by JASCO Corporation) Matrix-assisted laser desorption / ionization high-resolution mass spectrometer (MALDI-HRMS): JMS-S3000 (manufactured by JEOL Ltd.) High-performance liquid chromatography (HPLC): Nexera (manufactured by Shimadzu Corporation), e2695 (LC unit), 2489 (UV-Vis detector) (manufactured by Waters)

[0047] 2. Synthesis of Dimyristoyl Phosphatidylcholine (DMPC) Hyflo Super Cel (trademark) (3.18 g) was added to a 30 mL centrifuge tube, and a methanol solution containing choline glycerophosphate prepared with choline glycerophosphate (1.07 g, 4.12 mmol) and methanol (1.8 mL) was added. The solvent was removed and the mixture was dried for 2 days in a desiccator containing phosphorus oxide. The prepared choline glycerophosphate-Hyflo Super Cel complex, myristic acid (4.52 g, 19.8 mmol), DMAP (1.26 g, 10.3 mmol), DCC (4.08 g, 19.8 mmol), and super-dehydrated chloroform (80 mL) were added to a 200 mL round-bottom flask and heated and stirred at 30°C for 18 hours. At the end of the reaction, insoluble matter was removed by suction filtration, washed with chloroform, and the filtrate was concentrated. Chloroform (40 mL) was added to the concentration residue, precipitates were removed by filtration, and the solution was concentrated again and purified using a purification apparatus. The column used was an SI series (Size 60), and the solvent was chloroform / methanol = 100 / 0 to 50 / 50, vol / vol. In thin-layer chromatography (TLC), the fraction with Rf = 0.17 (developing solvent: chloroform / methanol / water = 70 / 26 / 4, vol / vol / vol) was concentrated, dried under reduced pressure, and then diisopropyl ether (40 mL) was added to the residue and stirred overnight at room temperature. After overnight, the solid dispersion solution was filtered, and the crystals on the filter paper were vacuum-dried to obtain the target product as a white powder. Yield: 1.74 g, yield 62%, δH (400 MHz. CDCl3): 5.24-5.14 (1H, m), 4.39 (1H, dd, J = 2.8, 11.9 Hz), 4.29 (2H, br), 4.12 (1H, dd, J = 7.3, 11.9 Hz), 4.00-3.85 (2H, m), 3.77 (2H, br), 3.34 (9H, s), 2.28 (4H, dt, J = 7.8, 8.2 Hz), 1.66-1.50 (4H, m), 1.37-1.19 (40 H, m), 0.88 (6H, t, J = 6.9 Hz).

[0048] 3. [EDMPC]+ [TfO] - Next, the synthesis of 1,2-dimyristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC) trifluoromethanesulfonate (TfO) (i.e., [EDMPC] + [TfO] - The following was the synthesis of ). Specifically, it was carried out as follows: First, DMPC (1.36 g, 2.0 mmol) and super-dehydrated chloroform (4 mL) were added to a dry 25 mL two-necked flask. While stirring this solution at room temperature, ethyl trifluoromethanesulfonate (258 μL, 2.0 mmol) was added in 129 μL increments. After the addition, the mixture was stirred at room temperature for 3 hours. After 3 hours, the reaction solution was transferred to a separatory funnel, washed with ethyl acetate (100 mL), and then saturated sodium bicarbonate aqueous solution (40 mL) was added, and the separatory funnel was shaken. The aqueous layer was removed, saturated saline solution (40 mL) was added, and the separatory funnel was shaken. The aqueous layer was removed, and the organic layer was separated, then sodium sulfate was added and allowed to stand for 10 minutes. After 10 minutes, sodium sulfate was removed by filtration, and the filtrate was concentrated. After concentration, the solution was purified using a purification apparatus. The column used was an SI series (Size 60), and the solvent was chloroform / methanol = 100 / 0 to 50 / 50, vol / vol. In TLC, the fraction with Rf = 0.34 (eluent: chloroform / methanol / water = 70 / 26 / 4, vol / vol / vol) was concentrated and vacuum-dried to obtain the target product as white crystals. Yield: 1.25 g, yield 73%, δH (400 MHz. CDCl3): 5.31-5.20 (1H, m), 4.58-4.44 (2H, m), 4.38-4.28 (1H, m), 4.28-4.10 (5H, m), 3.92-3.80 (2H, m), 3.32 (9H, s), 2.31 (4H, dt, J = 7.8, 8.7 Hz), 1.68-1.52 (4H, m), 1.47-1.15 (43 H, m), 0.88 (6H, t, J = 6.9 Hz).

[0049] 4. [EDMPC] + [Linoleic acid] -Add 100 mL of an anion exchange resin (Muromac XMA-413B-Cl) and 100 mL of pure water to a synthetic vial, and rotate and mix with a shaker (50 rpm) for 1 hour or more. The anion exchange resin has a cation represented by -CH2-N(CH3)3 + in its side chain.

[0050] Remove the supernatant, add about 100 mL of pure water again, and rotate and mix with a shaker (50 rpm) for 1 hour. Repeat this operation 8 more times. Remove the supernatant, add about 100 mL of a methanol / water (1 / 1, vol / vol) solution, and rotate and mix with a shaker (50 rpm) for 1 hour. Remove this supernatant, add about 100 mL of methanol, and rotate and mix with a shaker (50 rpm) for 1 hour. Repeat this operation 2 more times. Remove this supernatant, add about 100 mL of methanol, and immerse overnight. Pour 20 mL of the immersed anion exchange resin into a column and fill the column while gently tapping. Slowly pass a methanol / water (90 / 10, v / v) solution (200 mL) containing 2% sodium linoleate through the column filled with the anion exchange resin. Next, slowly pass methanol / water (90 / 10, v / v) (400 mL) through. Then, slowly pass ethanol (200 mL) through. Add [EDMPC] + [TfO] - 85.6 mg (0.1 mmoL) and EtOH (2.5 mL) to a vial to prepare an ethanol solution, and the prepared [EDMPC] + [TfO] -An ethanol solution was supported on an ion exchange resin. Ethanol was added onto the ion exchange resin, the flow rate was set to approximately 1.67 mL / min, and fractions of approximately 2.5 mL each were collected. The TLC of each fraction was checked, and the fractions with confirmed spots were mixed in a flask and concentrated. δH (400 MHz, CDCl3): 5.43 - 5.18 (4.2H, m), 4.57 - 4.47 (2.0H, br), 4.46 - 4.04 (8.5H, m), 3.54 - 3.34 (9.8H, m), 2.77 (1.6H, t, J = 6.9 Hz), 2.52 - 1.94 (13.6H, m), 1.68 - 1.50 (6.0H, m), 1.48 - 1.16 (61.4H, m), 0.96 - 0.83 (9.0H, m).

[0051] The following is [EDMPC] in this example + [Linoleic acid] - 's synthetic scheme.

[0052] [EDMPC] before ion exchange + [TfO] - and [EDMPC] after ion exchange + [Linoleic acid] - 's 1 H - NMR, 19 F - NMR, and 31 P - NMR were obtained according to the conventional method. The results were as shown in Figure 1. As shown in Figure 1, in the 1H - NMR peaks of [EDMPC] + [Linoleic acid] - after ion exchange, peaks derived from EDMPC and peaks derived from linoleic acid were observed. It was shown that the composition after ion exchange contained [EDMPC] + [Linoleic acid] - . Also, no peak was observed in the F - NMR of [EDMPC] + [Linoleic acid] - after ion exchange, indicating that the composition after ion exchange did not contain F atoms. From this, it can be seen that through ion exchange, [TfO] 19 was converted to [linoleic acid] - from -It was shown that ion exchange to was efficiently achieved.

[0053] As shown in Figure 2, the IR spectrum of the ion-exchanged composition shows peaks derived from EDMPC and linoleic acid anions, indicating that the ion-exchanged composition contains [EDMPC] + [Linoleic acid] - It was shown that it contains

[0054] As can be seen from Figures 1 and 2, [TfO] is obtained by ion exchange. - from [linoleic acid] - Efficient ion exchange was achieved.

[0055] Since peaks originating from the solvent (ethanol) remained in the composition after ion exchange in Figure 1, further vacuum drying was performed. This resulted in [EDMPC] + [Linoleic acid] - It was observed that ethyl groups were removed from some of the material, generating DMPC (see Figure 3). Further analysis by HPLC revealed the formation of ethyl linoleate. This suggests that the linoleate anion reacted with the ethyl group to form ethyl linoleate (see Figure 4).

[0056] Therefore, [EDMPC] after ion exchange + [Linoleic acid] - The ethanol solution was concentrated to approximately 5 mL and then diluted to 10 mL with ethanol. δH (600 MHz, CDCl3): 5.42-5.20 (5H, m), 4.57-4.47 (2H, br), 4.37-4.29 (1H, m), 4.26-4.10 (5H, m), 4.01-3.91 (2H, m), 2.77 (2H, t, J = 6.9 Hz), 2.38-2.26 (4H, m), 2.19-2.11 (2H, m), 2.05 (4H, quin, J = 6.9 Hz), 1.66-1.53 ​​(6H, m), 0.94-0.83 (9H, m).

[0057] Analysis by HPLC performed immediately after preparation showed no formation of ethyl linoleate, [EDMPC]+ [Linoleic acid] - It was not decomposed (see Figure 5). Also, [EDMPC] after ion exchange + [Linoleic acid] - HPLC analysis of the ethanol solution confirmed that the EDMPC-derived content was 6.91 mg / mL. Furthermore, the sample was dried under reduced pressure to obtain a dry weight of 9.68 mg per 1 mL. The theoretical value of EDMPC contained in 1 mg was calculated, and the purity was calculated using the following formulas 1 and 2, yielding a result of 99.2%. The purity calculation formulas are as follows:

[0058] Pure [EDMPC] + [Linoleic acid] - Theoretical value of EDMPC contained in = (Molecular weight of EDMPC skeleton 707.01) / ([EDMPC] + [Linoleic acid] - Molecular weight 985.45) ... (Equation 1) Purity (wt%) = Quantitative value of EDMPC (mg / mL) / Dry weight of 1 mL of solution (mg / mL) / Theoretical value of EDMPC × 100 ... (Equation 2)

[0059] In addition, even after being stored at -20°C for two months, [EDMPC] + [Linoleic acid] - It was stable (see Figure 6). [EDMPC] was stored in ethanol as shown in the HPLC chromatograms in Figures 5 and 6. + [Linoleic acid] - In this solution, the EDMPC peak was maintained, and the ethyl linoleate peak did not appear. Polyhydric alcohols such as propylene glycol and butylene glycol were used as the alcohol. HPLC analysis was performed four days after preparation. As a result, no degradation products observed during drying were detected, similar to those seen in the ethanol solution (see Figure 7). As shown in Figure 7, [EDMPC] was extracted with a polyhydric alcohol and prepared in a polyhydric alcohol. + [Linoleic acid] - Within this range, the EDMPC peak was maintained, while the ethyl linoleate peak did not appear.

[0060] 5. Complex formation between EDMPC and dodecyl sulfate

[0061] Next, dodecyl sulfate (DS) of 1,2-dimyristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC) (i.e., [EDMPC] + [DS] - ) was synthesized. Specifically, it was done as follows: First, [EDMPC] was placed in a glass vial. + [TfO] - (428 mg, 0.5 mmol) and ethyl acetate (50 mL) were added and mixed by rotation until a homogeneous solution was obtained. In a separate vial, sodium dodecyl sulfate (159 mg, 0.55 mmol) and Milli-Q (50 mL) were added and mixed by rotation until a homogeneous solution was obtained. These two solutions were placed in a 200 mL flask and stirred at room temperature for 1 hour. After stirring for 1 hour, the reaction solution was transferred to a separatory funnel, washed with ethyl acetate (100 mL), and allowed to stand at room temperature until the organic and aqueous layers separated. Next, the aqueous layer was removed, Milli-Q (40 mL) was added, and the separatory funnel was shaken. After standing at room temperature until the organic and aqueous layers separated, the aqueous layer was removed. This procedure was repeated one more time, the aqueous layer was removed, the organic layer was separated, sodium sulfate was added, and it was allowed to stand for 20 minutes. After 20 minutes, the sodium sulfate was removed by filtration, and the filtrate was concentrated. After concentration, the target product was obtained as white crystals by vacuum drying. Yield: 0.461 g, yield 95%, δH (400 MHz. CDCl3): 5.43-5.18 (1H, m), 4.57-4.47 (2H, m), 4.46-4.28 (1H, m), 4.28-4.04 (5H, m), 4.02-3.25 (2H, m), 3.38 (9H, s), 2.34 (4H, dt), 1.68-1.50 (6H, m), 1.48-1.16 (61 H, m), 0.89 (9H, t, J = 6.5 Hz). MALDI-HRMS m / z: [M-DS]+ Calcd for C38H77NO8P 706.5381, Found 706.5404. MALDI-HRMS m / z: [M-EDMPC] -Calcd for C12H25O4S 265.1479, Found 265.1471.

[0062] The following is the [EDMPC] in this embodiment. + [DS] - This is a synthesis scheme.

[0063]

[0064] [EDMPC] before ion exchange + [TfO] - and [EDMPC] after ion exchange + [DS] - of １ H-NMR, １９ F-NMR, and ３１ P-NMR was obtained according to standard procedures. The results are shown in Figure 8. As shown in Figure 1, [EDMPC] after ion exchange. + [DS] - of １ The 1H-NMR peaks showed peaks originating from EDMPC and peaks originating from DS, and the composition after ion exchange contained [EDMPC] + [DS] - It was shown that it contains [EDMPC] after ion exchange. + [DS] - of １９ No peaks were observed in F-NMR, indicating that the composition after ion exchange does not contain F atoms. Therefore, ion exchange removes [TfO] - From [DS] - It was shown that ion exchange to was efficiently achieved.

[0065] As shown in Figure 9, the molecular weight of the compound of the present invention was confirmed by MALDI-HRMS measurement. In MALDI-HRMS (positive mode, matrix: CHCA), the m / z was 706.5404 ([M-DS] + A peak corresponding to the calculated value (706.5381) was observed. In addition, in MALDI-HRMS (negative mode, matrix: CHCA), m / z 265.1471 ([M-EDMPC] -A peak matching the calculated value (265.1490) was observed.

[0066] As can be seen from Figures 8 and 9, [TfO] is obtained by ion exchange. - From [DS] - Efficient ion exchange was achieved.

[0067] Figure 10 shows [EDMPC] stored at -30°C for 8 months. + [DS] - of １ H-NMR and ３１ P-NMR was obtained according to standard procedures. [EDMPC] after storage + [DS] - of １ The H-NMR peaks showed peaks originating from EDMPC and peaks originating from DS. ３１ P-NMR showed the same peaks as immediately after synthesis, therefore [EDMPC] + [DS] - The structure remained stable.

[0068] [EDMPC] + [DS] - The dried product was prepared in an ethanol solution to a concentration of 10 mM, and analysis by HPLC confirmed that the content derived from the EDMPC skeleton was 10.1 mM (see Figure 11). From the aforementioned NMR and MALDI results, [EDMPC] + The counter ions are [DS] - Since this is clear, the purity was calculated from the quantitative results of EDMPC using formula 3 described below, and a result of 101% was obtained. The formula for calculating purity is as follows (formula 3).

[0069] Pure [EDMPC] + [DS] - Purity (wt%) contained in = Quantitative value of EDMPC (mM) / Theoretical value of EDMPC 10 mM × 100 ... (Equation 3)

[0070] Furthermore, as shown in Figure 12, the EDMPC skeleton was maintained even after storage in an ethanol solution at -20°C for 6 months.

Claims

1. A composition comprising a hydrophobic cation and a hydrophobic anion, wherein the hydrophobic cation and the hydrophobic anion together constitute 80% by weight or more of the total weight of hydrophobic molecules in the composition.

2. The composition according to claim 1, wherein the hydrophobic cation comprises a diglycerol group containing a higher fatty acid ester, and the hydrophobic anion is an anionic surfactant containing a medium-chain or long-chain carbon chain.

3. The composition according to claim 1 or 2, wherein the hydrophobic cation comprises 1,2-dimyristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC).

4. The composition according to any one of claims 1 to 3, wherein the hydrophobic anion comprises linoleic acid and / or dodecyl sulfate.

5. The composition according to any one of claims 1 to 4, further comprising a solvent, wherein the solvent comprises an alcohol.

6. The composition according to claim 5, wherein the alcohol comprises a monohydric lower alcohol.

7. The composition according to claim 5, wherein the alcohol comprises a polyhydric alcohol.

8. The composition according to any one of claims 1 to 7, wherein the hydrophobic cation comprises 1,2-dimyristoyl-sn-glycero-3-ethyl-phosphatidylcholine (EDMPC), the hydrophobic anion comprises an anionic surfactant containing a medium-chain or long-chain carbon chain, the solvent comprises an alcohol, and after storage at -20°C for two months, the total amount of EDMPC and the hydrophobic cation is 80% by weight or more of the dry weight of all hydrophobic molecules in the composition.

9. The composition according to claim 8, which does not contain dimyristoylphosphatidylcholine (DMPC) or contains DMPC but the DMPC content is less than 5% by weight.

10. The composition according to claim 9, wherein the DMPC content is 10% by weight or less after storage at -20°C for two months.

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