Zwitterion and sparingly soluble substance dissolving agent

Abstract

The present invention relates to a zwitterion which is represented by general formula (1) (where R1 represents a substituted or unsubstituted, linear or branched, saturated or unsaturated, C1-20 hydrocarbon group, a substituted or unsubstituted, saturated or unsaturated, C3-20 non-aromatic cyclic hydrocarbon group, a substituted or unsubstituted aromatic C6-30 hydrocarbon group, or a substituted or unsubstituted C7-31 aromatic hydrocarbon-alkyl group, A is the cation part of the zwitterion, and represents a cation selected from among imidazolium cations, phosphonium cations, ammonium cations, sulfonium cations, pyrazolium cations, pyridinium cations, pyrrolidinium cations, morpholinium cations, cyclopropenylium cations, and piperidinium cations, R2 represents a C1-4 alkylene group, each R3 represents the same or different C1-4 alkylene group, each R4 represents the same or different C1-4 alkylene group, m represents a number of 1-150, n represents a number of 0-10, B is the anion part of the zwitterion, and represents an anion selected from among -OSO3 -, -SO3 -, -COO-, -OP=O(H)O-, -OP=O(CH3)O-, -OP=O(OR5)O-, and -OP=O(OH)O-, and R5 is a C1-8 alkyl group which may have a heteroatom). The present invention also relates to a sparingly soluble substance dissolving agent and a cell cryopreservation agent which contain the zwitterion.

Description

Zwitterions and sparingly soluble substances solubilizers

[0001] The present invention relates to zwitterions and agents for solubilizing poorly soluble substances.

[0002] Despite possessing various physiological activities and other properties, poorly soluble substances have limited applicability. Dimethyl sulfoxide (DMSO) is often used as a solvent for such poorly soluble substances. However, some substances are insoluble in DMSO, and DMSO's toxicity is also a concern, necessitating the need for new solvents. DMSO is also used as a cryopreservation agent for cryopreserving cells. However, cryopreserved cells are known to cause hypertension, nausea, and vomiting when infused into recipients or when handled by cell handlers. Therefore, new cryopreservation agents for cells are needed. Kuroda et al. of Kanazawa University have developed several zwitterions as new, low-toxicity solvents (Patent Documents 1-3, Non-Patent Documents 1-4).

[0003] JP 2018-191623 A JP 2022-108914 A JP 2023-123780 A

[0004] Synthesis of N-oxyethylene substituted imidazolium-based zwitterions as a recyclable solvent for cellulose dissolution (Cellulose (2023) 30: 87-109) Cellulose dissolution using novel zwitterions with oligoethers (29th Annual Meeting of the Cellulose Society of Japan) Development of low-viscosity zwitterions with flexible spacers (12th Ionic Liquid Symposium) Development of low-viscosity zwitterions for biomass processing (2021 Chemical Society of Japan)

[0005] However, the zwitterions and liquid zwitterions described above are only capable of dissolving a limited number of substances and are toxic to cells, making them insufficiently practical. Therefore, an object of the present invention is to provide a substance that can dissolve a wide range of poorly soluble substances and has low cytotoxicity.

[0006] The present inventors have therefore investigated the chemical structures of the previously reported zwitterions, synthesized new zwitterions in which moderately hydrophilic ether chains have been introduced into the cation side chains of the zwitterions or into the spacer between the anion and cation. They then investigated their solubility for poorly soluble substances and found that a zwitterion represented by the following general formula (1) can dissolve substances that could not be dissolved by existing zwitterions or liquid zwitterions. Furthermore, they found that this zwitterion has low cytotoxicity and is useful as a cell cryopreservation agent.

[0007] That is, the present invention provides the following inventions [1] to [9]. [1] General formula (1)

[0008]

[0009] (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A is the cation portion of the zwitterion and represents a cation selected from an imidazolium cation, a phosphonium cation, an ammonium cation, a sulfonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, a cyclopropenylium cation, and a piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, and R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; m represents a number from 1 to 150; n represents a number from 0 to 10; B represents the anion portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR5 ) O - , and -OP=O(OH)O - R represents an anion selected from 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = CH2CH2, R 3 =CH2CH2, n=3, and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 1, A = imidazolium cation, R 2 = (CH2) 1,3,4または5 , n=0 and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2) 1,3または5 , n=0 and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 3, A = imidazolium cation, R 2 = (CH2) 3または5 , n=0 and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = SO3 - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = CH2CH2, n = 0, and B = SO3 - and a compound wherein R 1 = CH3, R 4 = CH2CH2, m = 2, A = ammonium cation, R 2 = (CH2)3, n = 0, and B = COO - (Excluding compounds in which R is a zwitterion represented by the formula: 1is a linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. [3] The zwitterion according to [1] or [2], wherein A is an imidazolium cation, an ammonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, or a piperidinium cation. [4] B is —COO - , -SO3 - , and -OP=O(OH)O - [5] The zwitterion according to any one of [1] to [4], which satisfies one or more conditions selected from the following (i) to (iii): (i) R 1 (ii) R is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. 4 is an alkylene group having 3 to 4 carbon atoms; (iii) m is a number of 4 or more and 150 or less. [6] General formula (1)

[0010]

[0011] (R 1represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A is the cation portion of the zwitterion and represents a cation selected from an imidazolium cation, a phosphonium cation, an ammonium cation, a sulfonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, a cyclopropenylium cation, and a piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, and R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; m represents a number from 1 to 150; n represents a number from 0 to 10; B represents the anion portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - R represents an anion selected from 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. 1 = CH3, R 4 = CH2CH2, m = 1 to 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = COO - and a compound wherein R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)5, n = 0, and B = COO -[7] The solubilizer for hardly soluble substances (excluding cellulose) according to [6], which satisfies one or more conditions selected from the following (i) to (iii): (i) R 1 (ii) R is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. 4 is an alkylene group having 3 to 4 carbon atoms; (iii) m represents a number from 4 to 150; [8] General formula (1)

[0012]

[0013] (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A is the cation portion of the zwitterion and represents a cation selected from an imidazolium cation, a phosphonium cation, an ammonium cation, a sulfonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, a cyclopropenylium cation, and a piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, and R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; m represents a number from 1 to 150; n represents a number from 0 to 10; B represents the anion portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR5 ) O - , and -OP=O(OH)O - R represents an anion selected from 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. 1 = CH3, R 4 = CH2CH2, m = 1 to 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)5, n = 0, and B = COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = SO3 - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = ammonium cation, R 2 = (CH2)3, n = 0, and B = COO - and a compound wherein R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = CH2CH2, n = 0, and B = SO3 - [9] The cell cryopreservation agent according to [8], which satisfies one or more conditions selected from the following (i) to (iii): (i) R 1 (ii) R is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. 4 is an alkylene group having 3 to 4 carbon atoms; (iii) m is a number from 4 to 150;

[0014] The zwitterions of the present invention have excellent solubility for a wide range of poorly soluble substances and are useful as solubilizers for various poorly soluble substances. In addition, the zwitterions of the present invention have low cytotoxicity and are useful as cryopreservation agents for cells.

[0015] 1 shows the results of a cytotoxicity test on mouse fibroblasts for OP3imC3C and OP3imC4S. 1 shows the results of a cytotoxicity test on mouse fibroblasts for OE9imC3C and OE9imC4S. 1 shows the results of a cytotoxicity test on mouse fibroblasts for OE2AnC2OE2OM1C and OE2MoC2OE2OM1C. 1 shows the results of a cryopreservation test on human breast cancer cells. 1 shows the results of a cryopreservation test on human kidney cells. 1 shows the results of a cryopreservation test on human lung cancer cells. 1 shows the results of a cryopreservation test on human kidney cells. 1 shows the results of a cryopreservation test on human lung cancer cells.

[0016] As used herein, a zwitterion refers to an ion having a zwitterionic structure. Here, a zwitterion is a molecule having both a positive charge and a negative charge within one molecule, and is also referred to as a zwitterion. As used herein, a poorly soluble substance refers to a substance that has low solubility in a solvent such as water. A solubilizer for poorly soluble substances refers to an agent that dissolves poorly soluble substances. Furthermore, as used herein, poorly soluble and poorly soluble are treated as synonyms. As used herein, a cell cryopreservation agent refers to an agent added when cryopreserving various cells, and is an additive for protecting cells during freezing and thawing.

[0017] One aspect of the present invention is a compound represented by general formula (1)

[0018]

[0019] (R 1represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A is the cation portion of the zwitterion and represents a cation selected from an imidazolium cation, a phosphonium cation, an ammonium cation, a sulfonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, a cyclopropenylium cation, and a piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, and R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; m represents a number from 1 to 150; n represents a number from 0 to 10; B represents the anion portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - R represents an anion selected from 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = CH2CH2, R 3 =CH2CH2, n=3, and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 1, A = imidazolium cation, R 2 = (CH2) 1,3,4または5 , n=0 and B=COO - a compound in which R 1= CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2) 1,3または5 , n=0 and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 3, A = imidazolium cation, R 2 = (CH2) 3または5 , n=0 and B=COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = SO3 - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = CH2CH2, n = 0, and B = SO3 - and a compound wherein R 1 = CH3, R 4 = CH2CH2, m = 2, A = ammonium cation, R 2 = (CH2)3, n = 0, and B = COO - (Excluding compounds in which:

[0020] The zwitterion of the present invention has a cationic portion represented by A and an anionic portion represented by B. The zwitterion of the present invention also has an ether chain ((OR 4 ) and (OR 3 The compound is characterized by the introduction of a cation side chain or a spacer between the anion and cation of the zwitterion. This structure is believed to significantly improve the solubility of a wide range of poorly soluble substances compared to existing zwitterion compounds.

[0021] R 1represents a substituted or unsubstituted, linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted, saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted, aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted, aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms.

[0022] "Substituted or unsubstituted" means that a hydrogen atom of a group may or may not be substituted with another group. Here, the substituent that can be substituted is not particularly limited. In addition, the number of substituents that can be further substituted for one substituent is not particularly limited, and can be appropriately selected in consideration of the desired glycol ether structure. In addition, in the above, substitution with the same substituent is not performed. In other words, an alkyl group as a substituent is not substituted with another alkyl group.

[0023] Examples of such substituents include halogen atoms such as fluorine atom, chlorine atom, bromine atom, and iodine atom; alkyl groups such as methyl group, ethyl group, tert-butyl group, and dodecyl group; aromatic hydrocarbon groups such as phenyl group, p-tolyl group, xylyl group, cumenyl group, naphthyl group, anthryl group, and phenanthryl group; alkoxy groups such as methoxy group, ethoxy group, and tert-butoxy group; aryloxy groups such as phenoxy group and p-tolyloxy group; alkoxycarbonyl groups such as methoxycarbonyl group, butoxycarbonyl group, octyloxycarbonyl group, and phenoxycarbonyl group; acyloxy groups such as acetoxy group, propionyloxy group, methacryloyloxy group, and benzoyloxy group; acetyl group, benzoyl group, isobutyryl group, acyloxy group, and the like. Examples of suitable groups include acyl groups such as acriloyl group, methacryloyl group, and methoxalyl group, alkylsulfanyl groups such as methylsulfanyl group and tert-butylsulfanyl group, arylsulfanyl groups such as phenylsulfanyl group and p-tolylsulfanyl group, alkylamino groups such as methylamino group and cyclohexylamino group, dialkylamino groups such as dimethylamino group, diethylamino group, morpholino group, and piperidino group, and arylamino groups such as phenylamino group and p-tolylamino group, as well as hydroxy group, carboxy group, formyl group, mercapto group, sulfo group, mesyl group, p-toluenesulfonyl group, amino group, nitro group, cyano group, trifluoromethyl group, trichloromethyl group, trimethylsilyl group, phosphinico group, and phosphono group. Among these, alkyl groups such as methyl group, ethyl group, tert-butyl group, and dodecyl group are preferred from the viewpoint of solubility.

[0024] As the linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a linear or branched, saturated or unsaturated hydrocarbon group having 1 to 16 carbon atoms is preferred, a linear or branched, saturated or unsaturated hydrocarbon group having 1 to 12 carbon atoms is more preferred, a linear or branched, saturated or unsaturated hydrocarbon group having 1 to 8 carbon atoms is even more preferred, and a linear or branched, saturated or unsaturated hydrocarbon group having 1 to 6 carbon atoms is even more preferred. Specific examples of the linear or branched saturated hydrocarbon group having 1 to 20 carbon atoms include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an n-pentyl group, an isoamyl group, a tert-pentyl group, a neopentyl group, an n-hexyl group, a 3-methylpentan-2-yl group, a 3-methylpentan-3-yl group, a 4-methylpentyl group, a 4-methylpentan-2-yl group, a 1,3-dimethylbutyl group, a 2-methyl ... butyl group, 3,3-dimethylbutyl group, 3,3-dimethylbutan-2-yl group, n-heptyl group, 1-methylhexyl group, 3-methylhexyl group, 4-methylhexyl group, 5-methylhexyl group, 1-ethylpentyl group, 1-(n-propyl)butyl group, 1,1-dimethylpentyl group, 1,4-dimethylpentyl group, 1,1-diethylpropyl group, 1,3,3-trimethylbutyl group, 1-ethyl-2,2-dimethylpropyl group, n-octyl group, 2-methylpentyl group, 1 ... 2,4-dimethylpentan-3-yl group, 1,1-dimethylpentan-1-yl group, 2,2-dimethylhexan-3-yl group, 2,3-dimethylhexan-2-yl group, 2,5-dimethylhexan-2-yl group, 2,5-dimethylhexan-3-yl group, 3,4-dimethylhexan-3-yl group, 3,5-dimethylhexan-3-yl group, 1-methylheptyl group, 2-methylheptyl group, 5-methylheptyl group, 2-methyl 1-ethylhexyl group, 2-ethylhexyl group, 1-propylpentyl group, 2-propylpentyl group, 1,1-dimethylhexyl group, 1,4-dimethylhexyl group, 1,5-dimethylhexyl group, 1-ethyl-1-methylpentyl group, 1-ethyl-4-methylpentyl group, 1,1,4-trimethylpentyl group, 2,4,4-trimethylpentyl group, 1-isopropyl-1,2-dimethylpropyl group, 1,1,3,3-tetramethylbutyl group, n-nonyl group, 1-methyloctyl group, 6-methyloctyl group, 1-ethylheptyl group, 1-(n-butyl)pentyl group, 4-methyl-1-(n-propyl)pentyl group, 1,5,5-trimethylhexyl group, 1,1,5-trimethylhexyl group, 2-methyloctan-3-yl group, n-decyl group , 1-methylnonyl group, 1-ethyloctyl group, 1-(n-butyl)hexyl group, 1,1-dimethyloctyl group, 3,7-dimethyloctyl group, n-undecyl group, 1-methyldecyl group, 1-ethylnonyl group, n-dodecyl group, n-tridecyl group, n-tetradecyl group, 1-methyltridecyl group, n-pentadecyl group, n-hexadecyl group, n-heptadecyl group, n-octadecyl group, n-nonadecyl group, etc. Among these, from the viewpoint of production, methyl group, ethyl group, isopropyl group, n-butyl group, isobutyl group, n-hexyl group, and 2-ethylhexyl group are preferred. From the viewpoint of toxicity, methyl group, ethyl group, isopropyl group, n-butyl group, and isobutyl group are even more preferred. Specific examples of linear or branched unsaturated hydrocarbon groups having 1 to 20 carbon atoms include vinyl, 1-propenyl, allyl, 1-butenyl, 3-butenyl, isoprenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, and heptadecenyl groups. , octadecenyl groups (for example, oleyl groups ((Z)-octadec-9-en-1-yl groups)), linoleoyl groups ((9Z,12Z)-octadeca-9,12-dienoyl groups), α-linolenoyl groups ((9Z,12Z,15Z)-octadeca-9,12,15-trienoyl groups), γ-linolenoyl groups ((6Z,9Z,12Z)-octadeca-6,9,12-trienoyl groups), and the like.

[0025] As the saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 12 carbon atoms is preferred, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 8 carbon atoms is more preferred, and a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 6 carbon atoms is even more preferred. Specific examples of saturated non-aromatic cyclic hydrocarbon groups having 3 to 20 carbon atoms include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl, norbornyl, and adamantyl groups, with cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups being preferred from the viewpoint of production. Specific examples of unsaturated non-aromatic cyclic hydrocarbon groups having 3 to 20 carbon atoms include cyclopropenyl, cyclobutenyl, cyclopentenyl, 2,4-cyclopentadienyl, cyclohexenyl, 2,5-cyclohexadienyl, and cyclooctenyl groups.

[0026] The aromatic hydrocarbon group having 6 to 30 carbon atoms is preferably an aromatic hydrocarbon group having 6 to 24 carbon atoms, more preferably an aromatic hydrocarbon group having 6 to 18 carbon atoms, and even more preferably an aromatic hydrocarbon group having 6 to 10 carbon atoms. Specific examples include a phenyl group, a biphenyl group, a 1-naphthyl group, a 2-naphthyl group, a 9-anthryl group, a 9-phenanthryl group, a 1-pyrenyl group, a 5-naphthacenyl group, a 1-indenyl group, a 2-azulenyl group, a 9-fluorenyl group, a terphenyl group, a quaterphenyl group, a mesityl group, a pentalenyl group, a binaphthalenyl group, a ternaphthalenyl group, a quaternaphthalenyl group, a heptalenyl group, a biphenylenyl group, an indacenyl group, a fluoranthenyl group, an acenaphthylenyl group, an aceanthryllenyl group, a phenalenyl group, a fluoranthenyl group, a phenanthryl ... Examples of the alkyl group include fluorenyl group, anthryl group, bianthracenyl group, teranthracenyl group, quaternanthracenyl group, anthraquinolyl group, phenanthryl group, triphenylenyl group, pyrenyl group, chrysenyl group, naphthacenyl group, pleiadenyl group, picenyl group, perylenyl group, pentaphenyl group, pentacenyl group, tetraphenylenyl group, hexaphenyl group, hexacenyl group, rubicenyl group, coronenyl group, trinaphthylenyl group, heptaphenyl group, heptacenyl group, pyrantrenyl group, and ovalenyl group. From the viewpoint of solubility, the phenyl group is preferable.

[0027] As the aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms, an aromatic hydrocarbon-alkyl group having 7 to 25 carbon atoms is preferred, an aromatic hydrocarbon-alkyl group having 7 to 19 carbon atoms is more preferred, and an aromatic hydrocarbon-alkyl group having 7 to 16 carbon atoms is even more preferred. Examples thereof include a phenyl-C1-6 alkyl group, a naphthyl-C1-6 alkyl group, and an anthracenyl-C1-6 alkyl group, and specific examples thereof include a benzyl group, a phenethyl group, a phenylpropyl group, a phenylbutyl group, a naphthylethyl group, a 3-phenylpropyl group, a 1-naphthylmethyl group, a 2-naphthylmethyl group, a 2-(1-naphthyl)ethyl group, a 2-(2-naphthyl)ethyl group, a 3-(1-naphthyl)propyl group, and a 3-(2-naphthyl)propyl group. Among these, a benzyl group is preferred from the viewpoint of solubility.

[0028] A is a cation moiety of the zwitterion and represents a cation selected from imidazolium cation, phosphonium cation, ammonium cation, sulfonium cation, pyrazolium cation, pyridinium cation, pyrrolidinium cation, morpholinium cation, cyclopropenylium cation, and piperidinium cation. Among these cations, imidazolium cation, ammonium cation, pyrazolium cation, pyridinium cation, pyrrolidinium cation, morpholinium cation, and piperidinium cation are preferred, imidazolium cation, ammonium cation, pyrazolium cation, pyridinium cation, pyrrolidinium cation, morpholinium cation, and piperidinium cation are more preferred, and imidazolium cation, ammonium cation, and morpholinium cation are even more preferred.

[0029] For example, specific examples of compounds of general formula (1) when A is an imidazolium cation include compounds represented by the following formula (1im): Substituents X may be the same or different and represent a hydrogen atom or a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 8 carbon atoms.

[0030]

[0031] Specific examples of the compound of general formula (1) when A is an ammonium cation or a morpholinium cation include compounds represented by the following formulae (1An) and (1Mo).

[0032]

[0033] In this way, the ether chain (OR 4 ) and (R 2 ) is considered to be attached to a heteroatom.

[0034] R 2 represents an alkylene group having 1 to 4 carbon atoms. 2Specific examples of the alkyl group include a methylene group, an ethylene group, a propylene group, a trimethylene group, a tetramethylene group, and a butylene group, with an ethylene group, a propylene group, and a butylene group being more preferred, and a propylene group and a butylene group being even more preferred. When the alkyl group has 3 or more carbon atoms, hydrophobicity is improved, which contributes to improving the solubility of poorly soluble substances.

[0035] R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms. 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms. 3 and R 4 Specific examples of the alkyl group include a methylene group, an ethylene group, a propylene group, a trimethylene group, a tetramethylene group, and a butylene group, with an ethylene group, a propylene group, and a butylene group being preferred, and a propylene group and a butylene group being more preferred. When the alkyl group has 3 or more carbon atoms, hydrophobicity is improved, which contributes to improving the solubility of poorly soluble substances.

[0036] Both m and n represent the average number of moles added, where m is a number from 1 to 150 and n is a number from 0 to 10. The average number of moles added refers to the average number of moles of oxyalkylene groups added per mole of zwitterion. The average number of moles added is a number average. m is preferably 3 to 150, more preferably 4 to 150, even more preferably 4 to 100, still more preferably 4 to 50, and even more preferably 4 to 9. When m is 4 or more, the viscosity of the zwitterion decreases, thereby contributing to improving the solubility of poorly soluble substances. n is more preferably 0 to 9, even more preferably 0 to 8, and even more preferably 0 to 6.

[0037] B is the anionic portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - Among these, -COO - , -SO3- , and -OP=O(OH)O - Preferably, the anion is selected from the group consisting of -COO - and -SO3 - More preferred are anions selected from:

[0038] R 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. 5 Examples of the alkyl group include a methyl group and an ethyl group.

[0039] Specific examples of the zwitterion represented by general formula (1) include the compounds shown in Table 1 below.

[0040]

[0041] The structural formulae of compounds 5, 13, and 14 shown in Table 1 are as follows:

[0042] The zwitterion represented by general formula (1) can be produced, for example, according to the following reaction scheme.

[0043]

[0044] (In the formula, R 6 represents an alkyl group or an aromatic hydrocarbon group, and R 7 represents an alkyl group, a halogenoalkyl group, or an aromatic hydrocarbon group; A, B, and R 1 - (OR 4 ) mR 2 , R 3 and n have the same meaning as above.

[0045] R 6 represents an alkyl group or an aromatic hydrocarbon group. Here, examples of the alkyl group include linear or branched alkyl groups having 1 to 8 carbon atoms. Specifically, methyl, ethyl, and tert-butyl groups are more preferred. Examples of the aromatic hydrocarbon group include a phenyl group, a halogenophenyl group, and a nitrophenyl group. R 7represents an alkyl group, a halogenoalkyl group, or an aromatic hydrocarbon group. Examples of alkyl groups include linear or branched alkyl groups having 1 to 8 carbon atoms. Specifically, methyl groups, ethyl groups, and the like are more preferred. Examples of halogenoalkyl groups include fluoroalkyl groups. Specifically, examples include trifluoromethyl groups. Examples of aromatic hydrocarbon groups include alkylphenyl groups. Specifically, examples include p-toluene groups.

[0046] Each step in the above reaction scheme will now be described. Step (1) is a step in which compound (2) is reacted with compound (3) to obtain compound (4). This step is a step in which the hydroxy group of compound (2) is sulfonylated. Compound (3) is a sulfonylating agent, and a sulfonyl halide compound such as tosyl chloride, mesyl chloride, or trifluoromethylsulfonyl chloride is preferably used. This reaction is preferably carried out in the presence of a base. Examples of the base that can be used include tertiary amines such as triethylamine and 4-dimethylaminopyridine, and inorganic bases such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, Na, and sodium hydride. The reaction can be carried out in a solvent at a temperature of 0°C to 100°C for approximately 1 hour to 40 hours. Examples of solvents that can be used include commonly used solvents such as halogenated hydrocarbons such as dichloromethane, aromatic hydrocarbons such as benzene and toluene, ethers, and acetonitrile.

[0047] Step (2) is a step in which compound (4) and compound (5) are reacted to obtain compound (6). This reaction is preferably carried out in the presence of a base. Examples of the base that can be used include tertiary amines such as triethylamine and 4-dimethylaminopyridine, and inorganic bases such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, Na, and sodium hydride. The reaction may be carried out in a solvent at a temperature of 0°C to 100°C for approximately 1 hour to 40 hours.

[0048] Step (3) is a step in which compound (6) and compound (7) are reacted to obtain compound (8). This reaction may be carried out in a solvent at a temperature of 0°C to 100°C for approximately 1 hour to 40 hours. The obtained compound (8) can be neutralized by adding an anion exchange resin and converted to zwitterion (1). As the anion exchange resin, an anion exchange resin having a quaternary ammonium group or the like can be used.

[0049] Zwitterions represented by general formula (1) include those that are liquid at room temperature and those that are solid at 100°C, and are stable without decomposition up to temperatures of 100°C or higher. Furthermore, they are low-toxic and have the ability to dissolve poorly soluble substances, making them useful as solubilizers for poorly soluble substances and cryopreservatives for cells. Solid zwitterions can be converted to liquid form, for example, by adding a small amount of water, and can be used as solubilizers for poorly soluble substances and cryopreservatives for cells, just like liquid zwitterions.

[0050] Another aspect of the present invention is a compound represented by general formula (1)

[0051]

[0052] (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A is the cation portion of the zwitterion and represents a cation selected from an imidazolium cation, a phosphonium cation, an ammonium cation, a sulfonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, a cyclopropenylium cation, and a piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, and R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; R 4are the same or different and represent an alkylene group having 1 to 4 carbon atoms; m represents a number from 1 to 150; n represents a number from 0 to 10; B represents the anion portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - R represents an anion selected from 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. 1 = CH3, R 4 = CH2CH2, m = 1 to 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = COO - and a compound wherein R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)5, n = 0, and B = COO - (Excluding compounds represented by the formula:) A solubilizer for hardly soluble substances (excluding cellulose) containing a zwitterion represented by the formula:

[0053] In the present invention, the term "slightly soluble substance" refers to a substance that does not dissolve at all in water or dissolves only slightly, and has a solubility in water (25°C) of 1% by weight or less, preferably 0.5% by weight or less, and particularly preferably 0.1% by weight or less. Examples of such slightly soluble substances include substances that are active ingredients in pharmaceuticals, veterinary drugs, quasi-drugs, cosmetics, and agricultural chemicals (including candidate substances that may become active ingredients), food additives, biological substances, and plant-derived substances (excluding cellulose), and also includes low-molecular-weight substances, as well as oligomers and polymers such as oligopeptides, polypeptides, polysaccharides, DNA, and RNA. "Slightly soluble pharmaceuticals" refer to drugs that are "slightly soluble," "slightly soluble," "extremely soluble," or "almost insoluble" as defined in the Japanese Pharmacopoeia, and specifically include antitumor agents, antibiotics, antihyperlipidemic agents, antibacterial agents, allergic disease treatments, hypertension treatments, arteriosclerosis treatments, blood circulation promoters, hormones, fat-soluble vitamins, diabetes treatments, antiandrogens, cardiotonic drugs, antiarrhythmic drugs, anti-inflammatory drugs, hypnotics, sedatives, tranquilizers, antiepileptic drugs, antidepressants, digestive system disease treatments, diuretics, local anesthetics, anticoagulants, antihistamines, etc. Examples of the poorly soluble substances include steroids, antimuscarinics, antimycobacterial agents, immunosuppressants, antithyroid drugs, antivirals, anxiolytic sedatives, astringents, β-adrenergic receptor blocking agents, cardiac inotropes, contrast media, corticosteroids, cough suppressants, diagnostic agents, diagnostic imaging agents, diuretics, dopaminergic agents, lipid-regulating agents, muscle relaxants, parasympathomimetics, thyroid calcitonin, prostaglandins, radiopharmaceuticals, sex hormones, stimulants, appetite suppressants, sympathomimetics, thyroid agents, vasodilators, isoflavones, xanthenes, etc. More specific examples of the poorly soluble substances include glycyrrhetinic acid and its salts, glycyrrhizinic acid and its salts, coumarin, ononin, liquiritin, peptides, polypeptides such as collagen, polysaccharides such as xylan, lignin, chloramphenicol, etc.

[0054] Examples of antitumor agents include methotrexate, taxol, doxorubicin hydrochloride, bleomycin hydrochloride, tamoxifen, cisplatin, carboplatin, cyclosporine, HER2 inhibitors, melphalan, dacarbazine, carmofur, enocitabine, etoposide, 5-fluorouracil, mitoxantrone, mesna, dimesna, aminoglutethimide, acrolein, cyclophosphamide, lomustine, carmustine, cyclophosphamide, busulfan, para-aminosalicylic acid, mercaptopurine, tegafur, azathioprine, vinblastine sulfate, mitomycin C, L-asparagine, ubenimex, paclitaxel, and dorsomorphin.

[0055] Examples of antibiotics include streptomycin, chloramphenicol, gentamicin, tetracycline, penicillin, amikacin, dibekacin, bacitracin, cephalexin, nystatin, erythromycin, fradiomycin sulfate, cefmetazole, and tolnaftate.

[0056] Examples of the antihyperlipidemic agent include cholestyramine, niceritrol, clinofibrate, clofibrate, fenofibrate, bezafibrate, soy sterol, tocopherol nicotinate, nicomol, probucol, simvastatin, colestimide, elastase, and the like.

[0057] Examples of antibacterial agents include chloramphenicol, rokitamycin, roxithromycin, cefatrizine, ofloxacin, ciprofloxacin hydrochloride, tosufloxacin tosylate, norfloxacin, lomefloxacin hydrochloride, pazufloxacin, cefpodoxime proxetil, midecamycin acetate, josamycin propionate, fosfomycin, and salts thereof.

[0058] Examples of therapeutic agents for allergic diseases include ebastine, mequitazine, methoxyphenamine, clemastine fumarate, cyproheptadine hydrochloride, fexofenadine hydrochloride, diphenhydramine, methdilamine, clemizole, and the like.

[0059] Examples of antihypertensive agents include nicardipine hydrochloride, delapril hydrochloride, barnidipine hydrochloride, efonidipine hydrochloride, benidipine hydrochloride, alacepril, captopril, cilnidipine, felodipine, amlodipine besylate, nisoldipine, manidipine hydrochloride, nitrendipine, nilvadipine, trandolapril, valsartan, candesartan cilexetil, urapidil, carvedilol, prazosin hydrochloride, bunazosin hydrochloride, doxazosin mesylate, reserpine, methyldopa, guanabenz acetate, deserpidine, meptame, meptamate, and the like.

[0060] Examples of therapeutic agents for arteriosclerosis include elastase, clofibrate, simfibrate, soy sterol, and nicomol.

[0061] Examples of therapeutic agents for osteoporosis include zoledronic acid.

[0062] Examples of blood circulation promoters include tocopherol acetate, tocopherol nicotinate, nicotinic acid benzyl ester, caffeine, tolazoline, verapamil, cyclandelate, and acetylcholine.

[0063] Hormonal agents include dexamethasone, dexamethasone acetate, betamethasone, betamethasone valerate, betamethasone dipropionate, beclomethasone propionate, prednisolone, prednisolone valerate, prednisolone acetate, methylprednisolone, methylprednisolone acetate, hydrocortisone, hydrocortisone acetate, hydrocortisone acetate propionate, amcinonide, triamcinolone, triamcinolone acetonide, fluocinolone acetonide, estriol, fluocinonide, hexestrol, methimazole, estriol propionate, clobetasone acetate, clobetasol propionate, and testosterone propionate. Examples of the steroid hormones that can be used include estradiol, testosterone enanthate, fluoxymesterone, stanolone, dromostanolone propionate, estradiol benzoate, estradiol propionate, estradiol valerate, ethinylestradiol, β-estradiol, mestranol, estriol acetate benzoate, fluorometholone, fludroxycortolone valerate, halcinonide, progesterone, hydroxyprogesterone caproate, pregnanediol, medroxyprogesterone acetate, dimethisterone, norethisterone, allylestrenol, gestonorone caproate, oxendolone, and insulin.

[0064] Examples of fat-soluble vitamin preparations include vitamin A, its precursor (β-carotene), vitamin D, vitamin E, vitamin K, and the like.

[0065] Examples of antidiabetic agents include gliclazide, tolbutamide, glibenclamide, troglitazone, epalrestat, buformin, metformin, and the like.

[0066] Examples of antiandrogens include gestonorone caproate, osapron acetate, flutamide, oxendolone, allylestrenol, chlormadinone acetate, and bicalutamide.

[0067] Examples of cardiotonic drugs include digoxin, digotoxin, and cobidecarenone.

[0068] Examples of antiarrhythmic drugs include lidocaine, bopindolol malonate, arotinolol hydrochloride, atenolol, pindolol, nadolol, propafenone hydrochloride, amiodarone hydrochloride, disopyramide, carteolol hydrochloride, and the like.

[0069] Anti-inflammatory agents include glycyrrhetinic acid, dipotassium glycyrrhizinate, aspirin, aluminum aspirin, ibuprofen, ketoprofen, ricyrrhetic acid, salicylic acid, acetaminophen, methyl salicylate, glycol salicylate, aminopyrine, phenacetin, mefenamic acid, flufenamic acid, aluminum flufenamate, tolfenamic acid, acemetacin, indomethacin, alclofenac, diclofenac, ibuprofen piconol, oxyphenbutazone, phenylbutazone, ketophenylbutazone, clofezone, tiaramide hydrochloride, diclofenac sodium, sulindac, naproxen, febufen, flurbiprofen, fenprofen, bufen Examples of such antihistamines include examac, mepirizole, perisoxal citrate, glafenine, bucolome, pentazocine, metiazinic acid, protizinic acid, pranoprofen, fenoprofen calcium, piroxicam, feprazone, fentiazac, bendazac, dimethylisopropylazulene, bufexamac, bucolome, benzydamine, tiaramide, tinoridine, ethenzamide, tenoxicam, chlortenoxicam, clidanac, naproxen, glycyrrhizin, azulene, camphor, thymol, 1-menthol, sazapyrine, alclofenac, diclofenac, suprofen, loxoprofen, diflunisal, tiaprofenic acid, oxaprozin, and felbinac.

[0070] Examples of hypnotic sedatives include barbital, amobarbital, amobarbital sodium, phenobarbital, phenobarbital sodium, secobarbital sodium, pentobarbital calcium, hexobarbital, triclophos, bromvaleryl urea, glutethimide, methaqualone, perlapine, nitrazepam, estazolam, flurazepam hydrochloride, flunitrazepam, estazolam, and the like.

[0071] Examples of tranquilizers include diazepam, lorazepam, and oxazolam.

[0072] Examples of antiepileptic drugs include phenytoin, carbamazepine, phenobarbital, primidone, phenacemide, ethylphenacemide, ethotoin, phensuximide, nitrazeban, clonazeban, and the like.

[0073] Examples of antidepressants include phenelzine, imipramine, and noxiptiline.

[0074] Examples of therapeutic agents for digestive system diseases include famotidine, sucralfate, aldioxa, irsogladine maleate, metoclopramide, cimetidine, omeprazole, lansoprazole, enprostil, gefarnate, teprenone, sulpiride, trepibutone, and oxethazaine.

[0075] Examples of diuretic drugs include spironolactone, chlorthalidone, polythiazide, triamterene, hydrochlorothiazide, and furosemide.

[0076] Examples of local anesthetics include ethyl aminobenzoate, procaine hydrochloride, lidocaine, dibucaine hydrochloride, tetracaine hydrochloride, benzyl alcohol, tecaine, bezocaine, pramoxine hydrochloride, catacaine hydrochloride, butanicaine hydrochloride, piperocaine hydrochloride, and chlorobutanol.

[0077] Anticoagulants include coumarin, heparin, and the like.

[0078] Examples of antiviral agents include acyclovir, nevirapine, zidovudine, zanamivir, oseltamivir, and favipiravir.

[0079] Examples of isoflavones include ononin, daidzein, biochanin A, glycitein, daidzin, glycitin, genistin, etc. Isoflavones may be either aglycones or glycosides.

[0080] Examples of poorly soluble substances used as quasi-drugs or cosmetics include dl-α-tocopherol acetate, α-tocopherol (vitamin E), methyl cinnamate, ethyl cinnamate, hexyl laurate, trichlorocarbanilide, triazine, benzophenone, triazole, eugenol, isoeugenol, ethyl methylphenylglycidate, geranyl acetate, piperonal, cinnamyl acetate, decyl oleate, terpenyl acetate, anilide, cinnamide, sulfonated benzimidazole, carotene, piroctone olamine, minoxidil, phytosteside, tocopherol nicotinate, ethinyl estradiol, polyporsterone, and ecdysteroids.

[0081] As the pesticide, any poorly soluble pesticide active ingredient having insecticidal, fungicidal, herbicidal, plant growth regulating, etc. can be used. For example, poorly soluble insecticides include abamectin, acrinathrin, amitraz, azadirachtin, azamethiphos, azinphos-methyl, azocyclotine, methiocarb, thiodicarb, trimethacarb, etofenprox, ethylthiometon, methoxychlor, chlorpyrifos-methyl, chlorfenson, chlorfluazuron, tebufenpyrad, bensultap, bifenthrin, bromopropylate, buprofezin, carbaryl, chlorfenapyr, clofentezene, coumaphos, diazinon, cycloprothrin, cyfluthrin, β-cyfluthrin, cypermethrin, α-cypermethrin, θ-cypermethrin, deltamethrin, diafenthiuron, dicofol, diflubenzuron, karin, kaspermethrin ... It is possible to dissolve bosulfan, endosulfan, esfenvalerate, etoxazole, fenazaquin, fenbutatin oxide, fenoxycarb, fenpyroximate, fipronil, fluazuron, flucycloxuron, flufenoxuron, flubendiamide, fenthion, halofenozide, hexaflumuron, hexythiazox, hydramethylnon, metaflumizone, lufenuron, milbemectin, novaluron, pentachlorophenol, pyridaben, rotenone, sulfluramide, tebufenozide, tebupirimfos, teflubenzuron, tetrachlorvinphos, tetradifon, benfuracarb, tolfenpyrad, triflumuron, tralomethrin, furathiocarb, and the like.

[0082] In addition, poorly soluble fungicides include bromuconazole, carpropamid, diclofen, metconazole, hexaconazole, fentin, mancozeb, maneb, diclomedine, azoxystrobin, isoprothiolane, benalaxyl, benomyl, bitertanol, captafol, captan, carbendazim, quinomethionate, chlorothalonil, chlozolinate, cyprodinil, dichlofluanid, dicloran, diclocymet, diethofencarb, dimethomorph, diniconazole, dithianon, tiadinil, epoxiconazole, famoxadone, fenarimol, fenbuconazole, fenfuram, fenpiclonil, fluazinam, fludioxonil, and fluoroisopropyl methylpropional. methylpropional, fluquinconazole, flusulfamide, flutolanil, folpet, hexachlorobenzene, imibenconazole, ipoconazole, iprodione, kresoxim-methyl, mepanipyrim, mepronil, metiram, nickel bis(dimethyldithiocarbamate), nuarimol, oxine copper, oxolinic acid, pencycuron, phthalide, procymidone, propineb, quintozene, sulfur, tebuconazole, tecloftalam, tecnazene, thifluzamide, thiophenate-methyl, thiram, tolclofos-methyl, triadimefon, tolylfluanid, triadimenol, triazoxide, triforine, triticonazole, vinclozolin, zineb, ziram, and the like.

[0083] Poorly soluble herbicides include bromobutide, aclonifen, clomethoxyfen, lactofen, prometryn, propazine, azafenidine, thenylchlor, bifenox, sulfentrazone, pyraflufen-ethyl, flumiclorac pentyl, flumioxazin, atrazine, indanofan, bensulfuron-methyl, benzofenap, bromofenoxime, chlorbromuron, chlorimuron-ethyl, chlornitrofen, chlorotoluron, chlorthal-dimethyl, clomeprop, dymron, desmedifam, dichlobenil, diflufenican, dimefuron, dinitramine, diuron, ethametsulfuron-methyl, triaziflam, fenoxaprop-ethyl, and furam. Examples of such an antibacterial agent include propmethyl, flazasulfuron, flumetsulam, fluthiacet-methyl, flupoxam, fluridone, flurtamone, oxaziclomefone, isoproturon, isoxaben, isoxapyrifop, lenacil, linuron, mefenacet, methabenzthiazuron, metobenzuron, naproanilide, nevron, norflurazon, oryzalin, oxadiazon, oxyfluorfen, phenmedipham, prodiamine, propyzamide, pyrazolinate, pyrazosulfuron-ethyl, pyributicarb, quinclorac, quizalofop-ethyl, rimsulfuron, siduron, simazine, terbuthylazine, terbutryn, thiazopyr, tracoxydim, and trietazine.

[0084] Examples of the sparingly soluble plant growth regulators include cyclanilide, flumetralin, 6-benzylaminopurine, forchlorfenuron, inabenfide, 2-(1-naphthyl)acetamide, paclobutrazol, N-phenylphthalamic acid, thidiazuron, and uniconazole.

[0085] Furthermore, examples of poorly soluble substances that are food additives include glycyrrhizin, L-ascorbic acid stearate, benzoic acid, isoeugenol, ergocalciferol (vitamin D2), eugenol, butyl parahydroxybenzoate, isopropyl parahydroxybenzoate, β-carotene, citronellyl formate, cholecalciferol (vitamin D3), propyl gallate, folic acid, lecithin, cinnamyl acetate, phenethyl acetate, ethyl cinnamate, dibutylhydroxytoluene, allyl hexanoate, methyl β-methyl ketone, riboflavin butyrate, and dl-α-tocopherol.

[0086] Examples of poorly soluble plant-derived substances include xylan, lignin, chondroitin sulfate, and glucomannan.

[0087] Examples of polypeptides include collagen, casein, albumin, elastin, and silk protein.

[0088] Examples of poorly soluble polysaccharides include chitin and chitosan.

[0089] It is speculated, but not limited to, that the solubilizer for poorly soluble substances of the present invention can dissolve various poorly soluble substances that have low polarity, i.e., tend to be hydrophobic, by introducing medium- or low-polarity groups into the zwitterion of the present invention. Furthermore, it is speculated, but not limited to, that the solubility of poorly soluble substances can be further improved by further increasing the proportion of medium- or low-polarity groups.

[0090] The zwitterion of the present invention further improves the solubility of a poorly soluble substance when it satisfies one or more of the following conditions (i) to (iii): (i) R 1 (ii) R is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. 4is an alkylene group having 3 to 4 carbon atoms; and (iii) m is a number of 4 or more and 150 or less. The reason why the solubility of the hardly soluble substance is improved is speculative and not limited, but it is thought that when any one or more of the above conditions (i) to (iii) are satisfied, hydrophobic interaction is improved.

[0091] The temperature at which poorly soluble substances can be dissolved is −10° C. to 100° C., furthermore, 10 to 80° C., and furthermore, 10 to 30° C. Generally, a higher temperature is better from the viewpoint of solubility, but some poorly soluble substances may lose their activity. Therefore, if the zwitterion of the present invention is used as a solubilizing agent, dissolution is possible without such high temperatures.

[0092] The amount of poorly soluble substance that the zwitterion of the present invention can dissolve is not particularly limited, but may be 0.00001 to 10 parts by mass, or even 0.0001 to 5 parts by mass, or even 0.001 to 1 part by mass, per 100 parts by mass of the zwitterion of the present invention. Because pharmaceuticals are often effective even at fairly low concentrations, even a small amount of poorly soluble substance dissolved in the zwitterion can be used as a pharmaceutical and provide the pharmaceutical effect, making it useful.

[0093] In addition to the zwitterions, the solubilizer for poorly soluble substances may contain additives suitable for the intended use of the poorly soluble substance, such as water, alcohols such as ethanol, polyols including diols and triols, sugars, salts, polymers such as polyethylene glycol and polyvinyl alcohol, other zwitterions, ionic liquids, zwitterionic polymers, ionic liquid polymers, amino acids, oils and fats, surfactants, and organic solvents such as methanol, dimethyl sulfoxide, and dimethylformamide that can be used in cell- or animal-related experiments.

[0094] In the agent for solubilizing poorly soluble substances of the present invention, in which the zwitterion is added with the additive, the concentration of the zwitterion is not particularly limited, but can be 0.1 to 100% by mass, preferably 1 to 90% by mass, more preferably 20 to 80% by mass, and even more preferably 30 to 70% by mass.

[0095] Another aspect of the present invention is a compound represented by general formula (1)

[0096]

[0097] (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A is the cation portion of the zwitterion and represents a cation selected from an imidazolium cation, a phosphonium cation, an ammonium cation, a sulfonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, a cyclopropenylium cation, and a piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, and R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms; m represents a number from 1 to 150; n represents a number from 0 to 10; B represents the anion portion of the zwitterion, -OSO3 - , -SO3 - , -COO - , -OP=O(H)O - , -OP=O(CH3)O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - R represents an anion selected from 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom.1 = CH3, R 4 = CH2CH2, m = 1 to 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)5, n = 0, and B = COO - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = (CH2)3, n = 0, and B = SO3 - a compound in which R 1 = CH3, R 4 = CH2CH2, m = 2, A = ammonium cation, R 2 = (CH2)3, n = 0, and B = COO - and a compound wherein R 1 = CH3, R 4 = CH2CH2, m = 2, A = imidazolium cation, R 2 = CH2CH2, n = 0, and B = SO3 - (Excluding compounds represented by the formula:

[0098] The origin of cells that can be cryopreserved is not particularly limited, and examples thereof include animal cells, insect cells, plant cells, yeast cells, and bacterial cells. Examples of animal cells include cells from humans, mice, rats, monkeys, pigs, dogs, sheep, and goats. Examples of bacteria include lactic acid bacteria, Escherichia coli, Bacillus subtilis, and cyanobacteria. The type of cell is also not particularly limited, and may be appropriately selected from the group consisting of pluripotent stem cells, tissue stem cells, somatic cells, and germ cells. Here, "pluripotent stem cells" is a general term for stem cells that have the ability to differentiate into cells of any tissue (pluripotency), and examples include embryonic stem cells (ES cells), induced pluripotent stem cells (iPS cells), embryonic germ stem cells (EG cells), and germ stem cells (GS cells). ES cells or iPS cells are preferred. Furthermore, "tissue stem cells" refer to stem cells whose differentiated cell lineages are limited to specific tissues but have the ability to differentiate into various cell types (pluripotency), such as hematopoietic stem cells in bone marrow, neural stem cells, hepatic stem cells, and skin stem cells. "Somatic cells" refer to cells that constitute multicellular organisms other than germ cells. Preferred examples include osteoclasts, fibroblasts, hepatocytes, pancreatic cells, muscle cells, bone cells, osteoblasts, chondrocytes, adipocytes, skin cells, pancreatic cells, kidney cells, lung cells, lymphocytes, erythrocytes, leukocytes, monocytes, and macrophages. "Germ cells" include gametes for sexual reproduction, i.e., eggs, oocytes, sperm, and sperm cells, as well as spores for asexual reproduction. The cells may be selected from the group consisting of sarcoma cells, established cell lines, and transformed cells. "Sarcoma" refers to cancer that develops in connective tissue cells derived from non-epithelial cells such as bone, cartilage, fat, muscle, and blood, and includes soft tissue sarcoma, malignant bone tumors, and the like. Sarcoma cells are cells derived from sarcoma. A "cell line" refers to a cultured cell that has been maintained outside the body for a long period of time, has certain stable properties, and can be subcultured semi-permanently. Examples of such cells include PC12 cells (derived from rat adrenal medulla), CHO cells (derived from Chinese hamster ovary), HEK293 cells (derived from human fetal kidney), HL-60 cells (derived from human white blood cells), and HeLa cells (derived from human cervical cancer)."Transformed cells" refer to cells whose genetic properties have been altered by introducing nucleic acid (e.g., DNA) from outside the cell. Transformation of animal cells, plant cells, and bacteria is carried out using conventionally known methods. Furthermore, when culturing ES cells or iPS cells, the cells may contain feeder cells, which are used as an auxiliary agent to create an environment necessary for cell proliferation and differentiation, as needed. Examples of feeder cells include mouse fibroblasts. These feeder cells can be treated in advance with gamma irradiation or antibiotics to prevent proliferation.

[0099] The concentration of the zwitterion of the present invention in the cell cryopreservation agent is preferably 0.1 to 70% by weight, more preferably 1 to 50%, and even more preferably 3 to 30% by weight, based on the total weight of the cell cryopreservation agent. In the cell cryopreservation agent of the present invention, the zwitterion of the present invention can prevent the formation of ice crystals and protect cells from physical destruction.

[0100] The cryopreservation agent of the present invention can also contain cell culture medium components in addition to the zwitterion of the present invention. In this case, the cryopreservation agent of the present invention functions as a medium composition or a culture composition. Examples of cell culture medium components include inorganic salts, buffers, carbohydrates, vitamins, proteins, peptides, fatty acids, lipids, trace elements, serum, hormones, growth factors, signal transduction substances, antibiotics, DMSO, etc.

[0101] When the cryopreservation agent of the present invention is used as a medium composition or a culture composition, the content of the zwitterion of the present invention is not limited, but is preferably 1 to 40% by mass, more preferably 3 to 25% by mass, and even more preferably 10 to 15% by mass. If the content of the zwitterion exceeds 40% by mass, the cell viability may decrease. However, the cell viability can be increased by using the slow freezing method or rapid freezing method described below.

[0102] The cryopreservation agent of the present invention can improve the viability of cells thawed after freezing a cell dispersion using slow or rapid freezing. The freezing conditions for cells in the slow freezing method can be appropriately set in accordance with conventional conditions. Specifically, slow freezing allows cells to be cooled to 0 to -200°C at a cooling rate of, for example, -0.1 to -15°C / min. Rapid freezing can be performed at a cooling rate of, for example, -15 to -20,000°C / min and a cooling temperature in the range of 0 to -200°C. A preferred method for thawing cryopreserved cells is to quickly transfer an ampoule containing the frozen cells to a 37°C water bath. The contents of the ampoule are transferred to a sterile tube using a pipette. Then, preheated medium supplemented with appropriate supplements is gradually added. The viable cell density is measured using trypan blue. An appropriate amount of the cell suspension is transferred to a flask and seeded at the cell density recommended in the cell line's data sheet.

[0103] The zwitterion used in the cell cryopreservation agent of the present invention preferably satisfies one or more of the following conditions (i) to (iii): (i) R 1 (ii) R is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. 4 is an alkylene group having 3 to 4 carbon atoms; and (iii) m is a number of 4 or more and 150 or less.

[0104] The present invention will now be described in more detail with reference to examples, but the present invention is not limited to these examples.

[0105] ( 1 H-NMR) 1 H-NMR was measured using an ECX400II (external magnetic field 400 MHz) manufactured by JEOL Ltd. The deuterated solvent used in the measurement was deuterated chloroform, deuterated DMSO, or deuterated water.

[0106] (Synthesis Example 1) (1) Synthesis of Zwitterion iBuOE2imC3C 250 g of diethylene glycol monoisobutyl ether (manufactured by Nippon Nyukazai Co., Ltd.), 700 g of toluene (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and 154 g of NaOH (manufactured by Tosoh Corporation) were mixed together. 352.2 g of p-tosyl chloride (manufactured by TCI) was added, and the mixture was stirred at 30°C for 2 hours. After the reaction, ion-exchanged water was added and the mixture was separated to obtain diethylene glycol monoisobutyl ether p-toluenesulfonate. 93.2 g of imidazole (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 932 g of tetrahydrofuran (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and 136.9 g of NaOH were mixed together. 433.2 g of diethylene glycol monoisobutyl ether p-toluenesulfonate was added, and the mixture was stirred at 60°C for 7 hours. The resulting reaction solution was filtered, and diethylene glycol monoisobutyl ether imidazole was obtained. 113.5 g of diethylene glycol monoisobutyl ether imidazole and 58.4 g of ethyl 4-bromobutyrate (manufactured by TCI) were mixed and refluxed for 6 hours at 80° C. After washing with diethyl ether, the mixture was mixed with an anion exchange resin, filtered, and the solvent was distilled off under reduced pressure to obtain iBuOE2imC3C. 1 H-NMR (400MHz, CHLOROFORM-D) δ 10.76 (s, 1H), 7.55 (s, 1H), 7.29 (s, 1H), 4.62-4.64 (m, 2H), 4.36-4.39 (m, 2H), 3.85-3.87 (m, 2H) ), 3.54-3.64 (m, 4H), 3.19-3.21 (dd, 2H), 2.14-2.24 (m, 4H), 1.80-1.90 (m1H), 0.88-0.90 (d, 6H)

[0107] (Synthesis Example 2) (2) Synthesis of zwitterion iBuOE2imC4S 89.0 g of diethylene glycol monoisobutyl ether imidazole obtained in the same manner as in Synthesis Example 1 and 62.9 g of 1,4-butane sultone (manufactured by TCI) were mixed and refluxed at 80° C. for 6 hours. After washing with diethyl ether, the solvent was distilled off under reduced pressure to obtain iBuOE2imC4S. 1H-NMR (400MHz, CHLOROFORM-D) δ 9.66 (s, 1H), 7.62 (s, 1H), 7.58 (s, 1H), 4.47-4.50 (m, 2H) , 4.35-4.37 (m, 2H), 3.85-3.87 (m, 2H), 3.62-3.65 (m, 2H) , 3.53-3.55 (m, 2H), 3.19-3.20 (d, 2H), 2.84-2.88 (m, 2H) , 2.08-2.12 (m, 2H), 1.81-1.89 (m, 3H), 0.88-0.89 (d, 6H)

[0108] Synthesis examples 1 and 2 were produced using the same compound as the one used in the synthesis. (3) Synthesis of OE9imC3C (Synthesis Example 3) 1 H-NMR (400MHz, DMSO-D6) δ 9.42 (s, 1H), 7.82 (s, 1H), 7.75 (s, 1H), 4.37-4.39 (m, 2H), 4.18-4.21 (m, 2 H), 3.78-3.80 (m, 2H), 3.41-3.56 (m, 32H), 3.24 (s, 3H), 1.87-1.95 (m, 4H)

[0109] (4) Synthesis of OE9imC4S (Synthesis Example 4) 1 H-NMR (400MHz, CHLOROFORM-D) δ 9.74 (s, 1H), 7.59 (s, 1H), 7.54 (s, 1H), 4. 44-4.51 (m, 2H), 4.34-4.37 (m, 2H), 3.50- 3.95 (m, 34H), 3.38 (s, 3H), 2.87-2.91 (m, 2H), 2.10-2.13 (m, 2H), 1.86-1.90 (m, 2H)

[0110] (5) Synthesis of OE4imC3C (Synthesis Example 5) 1 H-NMR (400MHz, CHLOROFORM-D) δ 10.47 (s, 1H), 7.58 (s, 1H), 7.37 (s, 1 H), 4.57-4.59 (m, 2H), 4.34-4.37 (m, 2H), 3.85-3.87(m, 2H), 3.47-3.72(m , 12H), 3.36 (s, 3H), 2.14-2.23 (m, 4H)

[0111] (6) Synthesis of OE4imC4S (Synthesis Example 6) 1H-NMR (400MHz, CHLOROFORM-D) δ 9.65 (s, 1H), 7.62 (s, 1H), 7.60 (s, 1H), 4.47-4 .49 (m, 2H), 4.33-4.36 (m, 2H), 3.85-3.87 (m, 2 H), 3.49-3.72 (m, 12H), 3.34 (s, 3H), 2.84-2.8 9 (t, 2H), 2.07-2.12 (m, 2H), 1.84-1.90 (m, 2H)

[0112] (7) Synthesis of OP3imC3C (Synthesis Example 7) 1 H-NMR (400MHz, CHLOROFORM-D) δ 10.51-10.58 (m, 1H), 7.55-7.71 (m, 1H), 7 .33-7.40 (m, 1H), 4.90-5.10 (m, 1H), 4.38- 4.41 (m, 4H), 3.29-3.88 (m, 9H), 2.14-2.2 6 (m, 4H), 1.59-1.62 (m, 3H), 1.06-1.14 (m, 6H)

[0113] (8) Synthesis of OP3imC4S (Synthesis Example 8) 1 H-NMR (400MHz, DMSO-D6) δ 9.24 (s, 1H), 7.87 (s, 1H), 7.81 (s, 1H), 4. 59-4.70 (m, 1H), 4.17-4.22 (m, 2H), 3.01-3 .83 (11H), 2.44-2.51 (m, 4H), 1.85-1.93 (m , 2H), 1.45-1.59 (m, 6H), 0.94-1.04 (m, 3H)

[0114] (9) Synthesis of BzOE1imC3C (Synthesis Example 9) 1 H-NMR (400MHz, DMSO-D6) δ 9.89 (s, 1H), 7.85 (s, 1H), 7.80 (s, 1H), 7.20-7.32 (m, 5H), 4.45-4. 48 (m, 4H), 4.18-4.20 (m, 2H), 3.73-3.77 (m, 2H), 1.82-1.93 (m, 4H)

[0115] (10) Synthesis of PhOE1imC3C (Synthesis Example 10) 1H-NMR (400MHz, DMSO-D6) δ 9.90 (s, 1H), 7.88 (s, 1H), 7.85 (s, 1H) , 7.22-7.26 (m, 2H), 6.88-6.94 (m, 3H) , 4.62-4.64 (m, 2H), 4.34-4.36 (m, 2H) , 4.18-4.21 (m, 2H), 1.82-1.94 (m, 4H)

[0116] (11) Synthesis of PhOE1imC4S (Synthesis Example 11) 1 H-NMR (400MHz, DMSO-D6) δ 9.30 (s, 1H), 7.83 (s, 1H), 7.80 (s, 1H), 7.25-7. 30 (m, 2H), 6.92-6.93 (m, 3H), 4.57-4.60 (m, 2H) , 4.34-4.36 (m, 2H), 4.19-4.22 (m, 2H), 2.43-2. 48 (m, 2H), 1.84-1.91 (m, 2H), 1.50-1.58 (m, 2H)

[0117] (12) Synthesis of OE2imC3C (Synthesis Example 12) The synthesis method is as described in Patent No. 7233669. 1 H-NMR (400MHz, DMSO-D6) δ 9.49 (s, 1H), 7.78 (s, 1H), 7.70 (s, 1H), 4 .29-4.35 (m, 2H), 4.10-4.18 (m, 2H), 3.70 -3.77 (m, 2H), 3.34-3.52 (m, 4H), 3.16 (s, 3H), 1.80-1.89 (m, 2H), 1.71-1.77 (m, 2H)

[0118] (13) Synthesis of OE2AnC2OE2OM1C (Synthesis Example 13) 500 g of dimethylaminoethoxyethanol (manufactured by Nippon Nyukazai Co., Ltd.), 700 g of toluene (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), 345 g of NaOH (manufactured by Tosoh Corporation), and 15 g of tetrabutylammonium bromide (manufactured by TCI) were added to a 3 L autoclave and mixed. 209 g of methyl chloride (manufactured by Shin-Etsu Chemical Co., Ltd.) was injected at 30°C over 2 hours, and the mixture was allowed to react for 24 hours. After the reaction, ion-exchanged water was added to perform layer separation, and the toluene layer was distilled to obtain N-methoxyethoxyethyl-N,N-dimethylamine. 20 g of 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxy)acetic acid (Sigma-Aldrich), 52 g of ethanol (Fujifilm Wako Pure Chemical Industries, Ltd.), and 0.2 g of concentrated sulfuric acid (Fujifilm Wako Pure Chemical Industries, Ltd.) were added and stirred at 80°C for 7 hours. Toluene and saturated brine were added to the resulting reaction solution, followed by layer separation. The resulting toluene layer was concentrated to obtain ethyl 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxyacetate. 5 g of N-methoxyethoxyethyl-N,N-dimethylamine and 13 g of ethyl 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxyacetate were mixed and refluxed at 80°C for 6 hours. After washing with diethyl ether, the mixture was mixed with an anion exchange resin, filtered, and the solvent was removed under reduced pressure to obtain OE2AnC2OE2OM1C. 1 H-NMR (400MHz, CHLOROFORM-D) δ 4.00-4.03 (m, 2H), 3.93-3.97 (m, 2H), 3.86-3.92 (m, 4H), 3.82-3.86 (m, 2H), 3.60-3.70 (m, 10H), 3.50-3.53 (m, 2H), 3.36-3.40 (s, 6H), 3.34-3.36 (s, 3H)

[0119] (14) Synthesis of OE2MoC2OE2OM1C (Synthesis Example 14) 18 g of morpholine (manufactured by Nippon Nyukazai Co., Ltd.), 21 g of NaOH (manufactured by Tosoh Corporation), and 53 g of tetrahydrofuran (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were mixed together. 40 g of 1-bromo-2-(2-methoxyethoxy)ethane (manufactured by TCI) was added dropwise at 60°C over 1 hour, and the mixture was allowed to react for 24 hours. The reaction solution was filtered, and the solvent was removed by distillation under reduced pressure to obtain N-methoxyethoxyethylmorpholine. 13 g of ethyl 2-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxyacetate obtained in the same manner as in Synthesis Example 13 and 5 g of N-methoxyethoxyethylmorpholine were mixed and refluxed at 80°C for 6 hours. After washing with diethyl ether, the mixture was mixed with an anion exchange resin, filtered, and the solvent was removed under reduced pressure to obtain OE2MoC2OE2OM1C. 1 H-NMR (400MHz, WATER-D2) δ 4.06-4.10 (m, 4H), 3.96-4.00 (m, 4H), 3.94-3.96 (s, 2H), 3.87-3.92 (m, 4H), 3.66-3.74 (m, 12H), 3.61-3.65 (m, 4H), 3.38-3.39 (s, 3H)

[0120] (Solubility Test) Zwitterion solutions prepared by adjusting the concentration of the zwitterion of the present invention (Examples 1 to 13) or OE2imC3C (Comparative Examples 1 and 2) with ion-exchange water to the concentrations shown in Tables 2 and 3, water, and DMSO were prepared, and the poorly soluble substances shown in Tables 2 and 3 were dissolved therein. Specifically, the mixture was stirred overnight at 30°C or 80°C, and immediately thereafter, the results were visually confirmed. The test was conducted by adjusting the concentration of the poorly soluble substance added to the zwitterion solution, water, or DMSO to the concentration (wt%) shown in Tables 2 and 3. Dissolution Criteria: ◯: Partial or complete dissolution was confirmed. ×: No dissolution was confirmed at all. The results are shown in Tables 2 and 3.

[0121]

[0122]

[0123] (Cytotoxicity Test) Cells (mouse fibroblasts: mNF) were cultured in a 96-well plate at 10,000 cells / well and then cultured for approximately 24 hours. A zwitterion solution was added to the medium (DMEM / FBS / penicillin-streptomycin / amphotericin B) so that the zwitterion concentration was 2 wt%, 5 wt%, or 10 wt%. The cells were cultured for 24 hours and then analyzed using CellTiter 96. TM Cell viability was investigated using Aqueous One Solution. Mouse normal fibroblasts (mNF) established from C57BL / 6-EGFP mice were used. As a result, as shown in Figures 1 to 3, the zwitterions of the present invention exhibited low cytotoxicity.

[0124] (Cryopreservation Test) (a) Preparation of Cryopreservation Solution Cryopreservation solutions having the following compositions were prepared. Cryopreservation solutions: CultureSure (registered trademark) cryopreservation solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) 10 wt% OP3imC3C (Synthesis Example 7) / H2O (ultrapure water) 10 wt% OP3imC4S (Synthesis Example 8) / H2O (ultrapure water) 10 wt% OE9imC3C (Synthesis Example 3) / H2O (ultrapure water) 10 wt% OE9imC4S (Synthesis Example 4) / H2O (ultrapure water) 10 wt% iBu-OE2imC4S (Synthesis Example 2) / H2O (ultrapure water) 10 wt% Synthesis of OE2AnC2OE2OM1C (Synthesis Example 13) / H2O (ultrapure water) Synthesis of 10 wt% OE2MoC2OE2OM1C (Synthesis Example 14) / H2O (ultrapure water)

[0125] (b) Cryopreservation of cells Cells to be frozen (human breast cancer cells: MDA-MB-231, human kidney cells: BOSC, human lung cancer cells: PC9) were treated with trypsin, centrifuged, and collected. The cells were then diluted with Dulbecco's modified Eagle's medium (DMEM) to measure the cell concentration. Subsequently, 1.0 × 10 cells were placed in a 1.5 ml tube. 6The cells were dispensed in 1 ml aliquots to obtain (*10^6) cells, centrifuged, and suspended in 100 μl of the above-mentioned cryopreservation solution. The cells were then frozen using a Mr. Frosty® cell freezing container at a cooling rate of -1°C / min and a cooling temperature of -85°C. Human breast cancer cells (MDA-MB-231) were obtained from Professor Erik Sahai (The Francis-Crick Institute, UK). Human kidney cells were obtained from Professor Erik Sahai at the Francis-Crick Institute, UK.

[0126] (c) Thawing of cells and counting of viable cell count One ml of medium was added to a cryopreservation vial, followed by thawing and centrifugation to remove the supernatant. Subsequently, the cells obtained by centrifugation were resuspended in medium, and the viable cell count was counted. The cell viability is the ratio (%) of the number of viable cells after thawing to the sum of the number of viable cells after thawing and the number of dead cells after freezing, as shown in the following formula. The cell recovery rate was calculated by multiplying the number of viable cells after thawing by 1.0 x 10 of the number of cells before freezing. 6 (*10 to the power of 6) and multiplied by 100.

[0127]

[0128] As shown in Figures 4 to 8, cells were observed to survive after freezing and thawing, confirming the cryoprotective effect. Furthermore, as shown in Figure 5, the cell survival rate after freezing and thawing exceeded 30%, and the cell recovery rate exceeded 15%, confirming the effectiveness of this agent as a cryopreservation agent.

Claims

1. General formula (1) 【Chemistry 1】 (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A represents a cation moiety of the zwitterion and is selected from the group consisting of imidazolium cation, phosphonium cation, ammonium cation, sulfonium cation, pyrazolium cation, pyridinium cation, pyrrolidinium cation, morpholinium cation, cyclopropenylium cation, and piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms, R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms, m is a number from 1 to 150, n represents a number from 0 to 10, B is the anionic portion of the zwitterion, -OSO 3 - , -SO 3 - , -COO - , -OP=O(H)O - , -OP=O(CH 3 ) O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - represents an anion selected from R 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. However, R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=imidazolium cation, R 2 =CH 2 CH 2 , R 3 =CH 2 CH 2 , n=3 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=1, A=imidazolium cation, R 2 = (CH 2 ) 1,3,4または5 , n=0 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=imidazolium cation, R 2 = (CH 2 ) 1,3または5 , n=0 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=3, A=imidazolium cation, R 2 = (CH 2 ) 3または5 , n=0 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=imidazolium cation, R 2 = (CH 2 ) 3 , n=0 and B=SO 3 - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=ammonium cation, R 2 = (CH 2 ) 3 , n=0 and B=COO - and Excluding compounds in which the substituent of R 1 is a (meth)acryloyl group or a (meth)acryloyloxy group. A zwitterion represented by

2. R 1 is a linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms.

3. 2. The zwitterion of claim 1, wherein A is an imidazolium cation, an ammonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, or a piperidinium cation.

4. B is -COO - , -SO 3 - , and -OP=O(OH)O - 2. The zwitterion of claim 1, wherein the anion is selected from the group consisting of:

5. The zwitterion according to any one of claims 1 to 4, which satisfies one or more conditions selected from the following (i) to (iii): (i) R 1 is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. (ii) R 4 is an alkylene group having 3 to 4 carbon atoms (iii) m is a number between 4 and 150.

6. General formula (1) 【Chemistry 2】 (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A represents a cation moiety of the zwitterion and is selected from the group consisting of imidazolium cation, phosphonium cation, ammonium cation, sulfonium cation, pyrazolium cation, pyridinium cation, pyrrolidinium cation, morpholinium cation, cyclopropenylium cation, and piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms, R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms, m is a number from 1 to 150, n represents a number from 0 to 10, B is the anionic portion of the zwitterion, -OSO 3 - , -SO 3 - , -COO - , -OP=O(H)O - , -OP=O(CH 3 ) O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - represents an anion selected from R 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. However, R 1 =CH 3 , R 4 =CH 2 CH 2 , m=1 to 2, A=imidazolium cation, R 2 = (CH 2 ) 3 , n=0 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=imidazolium cation, R 2 = (CH 2 ) 5 , n=0 and B=COO - and Excluding compounds in which the substituent of R 1 is a (meth)acryloyl group or a (meth)acryloyloxy group. A solubilizer for hardly soluble substances (excluding cellulose) containing a zwitterion represented by the formula:

7. The solubilizing agent for hardly soluble substances (excluding cellulose) according to claim 6, which satisfies one or more conditions selected from the following (i) to (iii): (i) R 1 is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. (ii) R 4 is an alkylene group having 3 to 4 carbon atoms (iii) m represents a number from 4 to 150

8. General formula (1) 【Transformation 3】 (R 1 represents a substituted or unsubstituted linear or branched, saturated or unsaturated hydrocarbon group having 1 to 20 carbon atoms, a substituted or unsubstituted saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic hydrocarbon group having 6 to 30 carbon atoms, or a substituted or unsubstituted aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms; A represents a cation moiety of the zwitterion and is selected from the group consisting of imidazolium cation, phosphonium cation, ammonium cation, sulfonium cation, pyrazolium cation, pyridinium cation, pyrrolidinium cation, morpholinium cation, cyclopropenylium cation, and piperidinium cation; R 2 represents an alkylene group having 1 to 4 carbon atoms, R 3 are the same or different and represent an alkylene group having 1 to 4 carbon atoms, R 4 are the same or different and represent an alkylene group having 1 to 4 carbon atoms, m is a number from 1 to 150, n represents a number from 0 to 10, B is the anionic portion of the zwitterion, -OSO 3 - , -SO 3 - , -COO - , -OP=O(H)O - , -OP=O(CH 3 ) O - , -OP=O(OR 5 ) O - , and -OP=O(OH)O - represents an anion selected from R 5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom. However, R 1 =CH 3 , R 4 =CH 2 CH 2 , m=1 to 2, A=imidazolium cation, R 2 = (CH 2 ) 3 , n=0 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=imidazolium cation, R 2 = (CH 2 ) 5 , n=0 and B=COO - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=imidazolium cation, R 2 = (CH 2 ) 3 , n=0 and B=SO 3 - a compound which is R 1 =CH 3 , R 4 =CH 2 CH 2 , m=2, A=ammonium cation, R 2 = (CH 2 ) 3 , n=0 and B=COO - and Excluding compounds in which the substituent of R 1 is a (meth)acryloyl group or a (meth)acryloyloxy group. A cell cryopreservation agent containing a zwitterion represented by the formula:

9. The cell cryopreservation agent according to claim 8, which satisfies one or more conditions selected from the following (i) to (iii): (i) R 1 is a linear or branched, saturated or unsaturated hydrocarbon group having 2 to 20 carbon atoms, a saturated or unsaturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms, an aromatic hydrocarbon group having 6 to 30 carbon atoms, or an aromatic hydrocarbon-alkyl group having 7 to 31 carbon atoms. (ii) R 4 is an alkylene group having 3 to 4 carbon atoms (iii) m represents a number from 4 to 150

Images