Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
  • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
  • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
  • C07D233/60—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with hydrocarbon radicals, substituted by oxygen or sulfur atoms, attached to ring nitrogen atoms
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/04—Preserving or maintaining viable microorganisms

Definitions

• the present invention relates to zwitterions and agents for dissolving poorly soluble substances.
• 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.
• [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.
• A is an imidazolium cation, an ammonium cation, a pyrazolium cation, a pyridinium cation, a pyrrolidinium cation, a morpholinium cation, or a piperidinium cation.
• B is an anion selected from —COO ⁇ , —SO 3 ⁇ , and —OP ⁇ O(OH)O ⁇ .
• zwitterion refers to an ion having a zwitterionic structure.
• a zwitterion is a molecule having both a positive charge and a negative charge in one molecule, and is also called a zwitterion.
• a poorly soluble substance refers to a substance that has low solubility in a solvent such as water.
• a solubilizer for a poorly soluble substance refers to an agent that dissolves a poorly soluble substance.
• poorly soluble and poorly soluble are treated as synonyms.
• a cell cryopreservation agent is an agent that is added when cryopreserving various cells, and is an additive that protects cells during freezing and thawing.
• One aspect of the present invention is a compound represented by general formula (1):
• the zwitterion of the present invention has a cationic moiety represented by A and an anionic moiety represented by B.
• the zwitterion of the present invention is characterized by the introduction of moderately hydrophilic ether chains ((OR 4 ) and (OR 3 )) into the cationic side chain of the zwitterion or into the spacer between the anion and the cation. This structure is believed to significantly improve the solubility of a wide range of poorly soluble substances compared to existing zwitterionic compounds.
• R1 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.
• substituted or unsubstituted means that a hydrogen atom of a group may or may not be substituted with another group.
• the substituents that may be substituted are not particularly limited.
• the number of substituents to be further substituted for one substituent is not particularly limited, and can be appropriately selected in consideration of the desired glycol ether structure. Note that, 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.
• n-octyl 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-methylhexan-2-yl group, 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-methylh
• methyl, ethyl, isopropyl, n-butyl, isobutyl, n-hexyl, and 2-ethylhexyl are preferred, and from the viewpoint of toxicity, methyl, ethyl, isopropyl, n-butyl, and isobutyl are even more preferred.
• 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), ⁇ -linolenoyl groups ((9Z,12Z,15Z)-octadeca-9,12,15-trienoyl groups), ⁇ -linolenoyl groups ((6Z,9Z,12Z)-octadeca-6,9,12-trienoyl groups), and the like.
• octadecenyl groups for example, oleyl groups ((Z)-octadec-9-en-1-yl groups)
• linoleoyl groups ((9Z,12Z)-octadeca-9,12-dienoyl groups)
• 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.
• saturated non-aromatic cyclic hydrocarbon group having 3 to 20 carbon atoms include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cyclooctyl group, a norbornyl group, and an adamantyl group. From the viewpoint of production, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group are preferred.
• 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.
• aromatic hydrocarbon group having 6 to 30 carbon atoms an aromatic hydrocarbon group having 6 to 24 carbon atoms is preferred, an aromatic hydrocarbon group having 6 to 18 carbon atoms is more preferred, and an aromatic hydrocarbon group having 6 to 10 carbon atoms is even more preferred.
• 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
• Aromatic hydrocarbon-alkyl groups having 7 to 31 carbon atoms are preferably aromatic hydrocarbon-alkyl groups having 7 to 25 carbon atoms, more preferably aromatic hydrocarbon-alkyl groups having 7 to 19 carbon atoms, and even more preferably aromatic hydrocarbon-alkyl groups having 7 to 16 carbon atoms. Examples include phenyl-C1-6 alkyl groups, naphthyl-C1-6 alkyl groups, and anthracenyl-C1-6 alkyl groups.
• A is the cation portion 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.
• R5 represents an alkyl group having 1 to 8 carbon atoms which may have a heteroatom, and examples of R5 include a methyl group and an ethyl group.
• the zwitterion represented by general formula (1) can be produced, for example, according to the following reaction scheme:
• R6 represents an alkyl group or an aromatic hydrocarbon group.
• the alkyl group include linear or branched alkyl groups having 1 to 8 carbon atoms. Specifically, a methyl group, an ethyl group, or a tert-butyl group is more preferred.
• the aromatic hydrocarbon group include a phenyl group, a halogenophenyl group, and a nitrophenyl group.
• R7 represents an alkyl group, a halogenoalkyl group, or an aromatic hydrocarbon group.
• the alkyl group include linear or branched alkyl groups having 1 to 8 carbon atoms. Specifically, methyl groups, ethyl groups, and the like are more preferred.
• the halogenoalkyl group include fluoroalkyl groups. Specifically, examples include trifluoromethyl groups.
• the aromatic hydrocarbon group include alkylphenyl groups. Specifically, examples include p-toluene groups.
• This reaction is preferably carried out in the presence of a base, such as a tertiary amine, triethylamine, 4-dimethylaminopyridine, or an inorganic base, such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, Na, or sodium hydride.
• a base such as a tertiary amine, triethylamine, 4-dimethylaminopyridine, or an inorganic base, such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, Na, or sodium hydride.
• the reaction may be carried out in a solvent at a temperature of 0° C. to 100° C. for about 1 hour to 40 hours.
• a general-purpose solvent such as a halogenated hydrocarbon such as dichloromethane, an aromatic hydrocarbon such as benzene or toluene, an ether, or acetonitrile can be used.
• Step (2) is a step in which compound (4) is reacted with compound (5) to obtain compound (6).
• This reaction is preferably carried out in the presence of a base.
• the base 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 about 1 hour to 40 hours.
• 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 above 100°C. Furthermore, they are low in toxicity and have the ability to dissolve poorly soluble substances, making them useful as solubilizers for poorly soluble substances and as cryopreservatives for cells. Solid zwitterions can also be made liquid by adding a small amount of water, for example, and can be used, like liquid zwitterions, as solubilizers for poorly soluble substances and as cryopreservatives for cells.
• Another aspect of the present invention is a compound represented by general formula (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; R2 represents an alkylene group having 1 to
• 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.
• 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.
• Antineoplastic 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.
• Antiviral agents include acyclovir, nevirapine, zidovudine, zanamivir, oseltamivir, and favipiravir.
• isoflavones examples include ononin, daidzein, biochanin A, glycitein, daidzin, glycitin, and genistin.
• Isoflavones may be either aglycones or glycosides.
• Slightly soluble substances used in quasi-drugs or cosmetics include dl- ⁇ -tocopherol acetate, ⁇ -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, anilides, cinnamide, sulfonated benzimidazole, carotene, piroctone olamine, minoxidil, phytosteside, tocopherol nicotinate, ethinyl estradiol, polyporsterone, and ecdysteroids.
• 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, fluoroquinol ...
• Examples include imide, 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, etc.
• Slightly 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 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.
• polypeptides examples include collagen, casein, albumin, elastin, silk protein, etc.
• 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; R2 represents an alkylene group having 1 to
• “Sarcoma” refers to cancers that develop in connective tissue cells derived from non-epithelial cells, such as bone, cartilage, fat, muscle, and blood, and includes soft tissue sarcoma and malignant bone tumors.
• Sarcoma cells are cells derived from sarcoma.
• Cell lines refer to cultured cells that have been maintained outside the body for a long period of time, have certain stable properties, and can be subcultured semi-permanently. Examples 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 leukocytes), 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 performed using conventionally known methods. Furthermore, when culturing ES cells or iPS cells, feeder cells may be optionally included as supplementary cells used to create an environment necessary for cell proliferation and differentiation. Examples of feeder cells include mouse fibroblasts. These feeder cells can be pretreated with gamma irradiation or antibiotics to prevent proliferation.
• nucleic acid e.g., DNA
• 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% based on the total weight of the cell cryopreservation agent.
• the zwitterion of the present invention can prevent the formation of ice crystals and protect cells from physical destruction.
• the contents of the ampoule are pipetted into a sterile tube. Pre-warmed medium with appropriate supplements is then slowly added. 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 on the cell line data sheet.
• 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.
• (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. The cells obtained by centrifugation were then resuspended in medium and the viable cell count was counted.
• the cell viability is the percentage (%) of the viable cell count after thawing to the sum of the viable cell count after thawing and the dead cell count after freezing, as shown in the following formula.
• the cell recovery rate is calculated by dividing the viable cell count after thawing by the cell count before freezing, 1.0 x 10 6 (*10 6 ), and multiplying the result by 100.

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• 2025
  • 2025-02-07 WO PCT/JP2025/004239 patent/WO2025170074A1/ja active Pending
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