Squalene derivatives, cation transporters, and anti-inflammatory agents

Abstract

To provide a novel squalene derivative, a cation transporter, and an anti-inflammatory agent.SOLUTION: The present invention provides a squalene derivative represented by a formula I, where n is an integer of 1 or more and 5 or less.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to squalene derivatives, cation transport agents, and anti-inflammatory agents. [Background technology]

[0002] Squalene is a type of triterpene obtained from shark liver oil and algae, and is known to have various physiological activities, including antioxidant effects, protective effects against gamma radiation, and antitumor effects.

[0003] Patent Document 1 describes that squalene has the effect of increasing total cholesterol in the blood of patients with hypocholesterolemia.

[0004] Patent Document 2 describes the anticancer and cancer metastasis-inhibiting effects of an ester of squalene-ω-alcohol and vitamin A acid, which possesses both the anticancer effects of vitamin A and its derivatives and the cancer metastasis-inhibiting effects of squalene and its derivatives. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Japanese Patent Publication No. 2008-266306 [Patent Document 2] Special Publication No. 60-38370 [Overview of the project] [Problems that the invention aims to solve]

[0006] The object of this invention is to provide novel squalene derivatives, cation transporters, and anti-inflammatory agents. [Means for solving the problem]

[0007] The present invention includes the following embodiments. [1] A squalene derivative represented by the following formula (I). [Chemical formula] However, in formula (I), n is an integer of 1 or more and 5 or less. [2] The squalene derivative according to [1], wherein n in the formula (I) is 2 or 3. [3] A cation transporter containing the squalene derivative according to [1] or [2] as an active ingredient. [4] The cation transporter according to [3], which is used in a cation membrane separation system. [5] The cation transporter according to [3] or [4] for transporting at least one metal ion selected from the group consisting of alkali metal ions, alkaline earth metal ions, iron group metal ions, platinum group metal ions, and actinide series metal ions. [6] An anti-inflammatory agent containing the squalene derivative according to [1] or [2] as an active ingredient. [7] A food or drink containing the squalene derivative according to [1] or [2]. [8] A medicine containing the squalene derivative according to [1] or [2]. [Advantages of the Invention]

[0008] According to the present invention, a novel squalene derivative, a cation transporter, and an anti-inflammatory agent can be provided. [Brief Description of the Drawings]

[0009] [Figure 1] 1H-NMR spectrum of Experimental Example 3. [Figure 2] 1H-NMR spectrum of Experimental Example 4. [Figure 3] Graph showing the number of GFP-expressing cells for each added concentration in Experimental Example 5. [Figure 4] Graph showing the GFP-expressing area for each added concentration in Experimental Example 5. [Figure 5] Graph showing the GFP-expressing intensity for each added concentration in Experimental Example 5. [Embodiments for Carrying Out the Invention]

[0010] Embodiments of the present invention will be described in detail below. However, the present invention is not limited to the embodiments described later, and various modifications are possible as long as they do not depart from the spirit of the invention. In the specification and claims, when a numerical range is expressed using "~", that numerical range shall include the numbers on both sides of "~".

[0011] [Squalene derivatives] The squalene derivative of the present invention is a compound represented by the following formula (I).

[0012] [ka]

[0013] In formula (I), n is an integer from 1 to 5, and the type of ion selectively retained can be changed depending on n. Preferably, n=1 when retaining lithium ions, n=2 when retaining sodium ions, n=3 when retaining potassium ions, n=4 when retaining ammonium ions, and n=5 when retaining barium ions. The squalene derivative of the present invention has excellent anti-inflammatory properties.

[0014] The squalene derivatives of the present invention can be synthesized by starting with squalene (a compound represented by the following formula (1)) to synthesize 2,3-oxidesqualene (a compound represented by the following formula (2); also called 2,3-epoxysqualene), and then substituting the 2 position of 2,3-oxidesqualene with 12-crown-4-ether (when n=1), 15-crown-5-ether (when n=2), 18-crown-6-ether (when n=3), 21-crown-7-ether (when n=4), or 24-crown-8-ether (when n=5) (hereinafter, these crown ethers will be collectively referred to as (3×m)-crown-m-ethers, where m=n+3).

[0015] [ka]

[0016] First, we will describe a method for synthesizing 2,3-oxidesqualene using squalene as a raw material. A method for synthesizing 2,3-oxidesqualene by epoxidizing the terminals of squalene is disclosed, for example, in EE van Tamelen, TJ Curphey, “The selective in vivo oxidation of the terminal double bonds in squalene”, Tetrahedron Letters, 3, 1962, 121-124. The method described below is just one example, and conventionally known methods can be used to synthesize 2,3-oxidesqualene from squalene. Squalene can be obtained from commercially available products, for example. Dissolve 2 equivalents of N-bromosuccinimide in aqueous ethylene glycol dimethyl ether with squalene and stir at room temperature (23°C) for 1 hour. Concentrate the reaction solution and purify it using silica gel chromatography to obtain 2,3-bromohydrin squalene. Stir the obtained 2,3-bromohydrin squalene in a basic ethanol solution at room temperature for 1 hour. Concentrate the reaction solution and purify it using silica gel chromatography with a hexane fraction containing 5% ethyl acetate to obtain 2,3-oxidesqualene.

[0017] Next, the 2-position of the synthesized 2,3-oxidesqualene is reacted with a nucleophile such as 2-hydroxymethyl-12-crown-4-ether or 2-hydroxymethyl-15-crown-5-ether by the following method to obtain a novel squalene derivative represented by the following formula (I). In formula (I), n is an integer between 1 and 5.

[0018] [ka]

[0019] The present invention provides a method for producing a squalene derivative by dissolving 2,3-oxidesqualene in an alcohol such as 2-propanol, adding 2-hydroxymethyl-12-crown-4-ether or 2-hydroxymethyl-15-crown-5-ether, or other 2-hydroxymethyl-(3×(n+3))-crown-(n+3)-crown ether (where n is an integer from 1 to 5), and heating to approximately 80°C to cause a reaction. After cooling, water is added, followed by extraction with ethyl acetate, and then purification by column chromatography to isolate the squalene derivative of the present invention.

[0020] The solvent used to dissolve 2,3-oxidesqualene can be, for example, acetone, 1,4-dioxane, tetrahydrofuran, tert-butyl alcohol, dimethylformamide, or chloroform. The amount of 2-hydroxymethyl-(3×(n+3))-crown-(n+3)-ether added is preferably in the range of 50 to 500 equivalents relative to 2,3-oxidesqualene. The reaction temperature is preferably in the range of 50 to 90°C, and the reaction time is preferably in the range of 4 to 9 hours.

[0021] The squalene derivative of the present invention is useful as a cation transport agent, as will be described later. It is also useful as an active ingredient in anti-inflammatory agents, as will be described later.

[0022] [Cation transport agent] Currently, the separation of cations such as metal ions is mainly done using extractants, but this method requires an amount of extractant equal to or greater than the amount of cations being treated. Therefore, while it is effective when the amount of cations to be treated is small, when considering the separation and removal of radioactive cations, for example, the amount of treatment is large, requiring the synthesis of large amounts of extractant, making it inefficient. In fact, many cationic extractants have been developed and are used for the separation, concentration, and removal of cations, but these extractants require an equivalent amount or greater than the amount of cations. Therefore, it is necessary to synthesize large amounts of extractant, which is problematic as it consumes large amounts of resources and a lot of energy for synthesis. As an alternative to treatment methods using cationic extractants, treatment with adsorbents is also being considered, but this method has the problem that continuous steady operation is not possible because regeneration is required each time, and in particular when used for the treatment of radioactive waste liquid, regeneration is difficult and it is not practical. Therefore, the development of new technologies that can process large quantities of cations continuously and with minimal resources or energy, as an alternative to the current methods of separating cations using extractants and adsorbents, is highly anticipated and important.

[0023] The inventors of the present invention have found that the squalene derivative represented by the above formula (I) and the cation exhibit an association characteristic of 1:1 molar ratio and are stably maintained for a long period of time of 6 months or more, leading to the invention of the cation transport agent of the present invention.

[0024] The cation transport agent of the present invention contains a squalene derivative represented by formula (I) as an active ingredient. Here, "as an active ingredient" means that it primarily functions as a cation transporter, that is, it mainly performs the function of adsorbing and desorbing cations.

[0025] The squalene derivative represented by formula (I) is preferably one in which n=2 or 3.

[0026] The cation is not particularly limited, but at least one metal ion selected from the group consisting of alkali metal ions, alkaline earth metal ions, iron group metal ions, platinum group metal ions, and actinide series metal ions is preferable, alkali metal ions are more preferable, and sodium ions or potassium ions are even more preferable. Examples of the alkali metal ions include lithium ions (Li + ), sodium ions (Na + ), potassium ions (K + ), rubidium ions (Rb + ), cesium ions (Cs + ), and francium ions (Fr + ). Examples of the alkaline earth metal ions include calcium ions (Ca 2+ ), strontium ions (Sr 2+ ), barium ions (Ba 2+ ), radium ions (Ra 2+ ), beryllium ions (Be 2+ ), and magnesium ions (Mg 2+ ). Examples of the iron group metal ions include iron(II) ions (Fe 2+ ), iron(III) ions (Fe 3+ ), cobalt(II) ions (Co 2+ ), cobalt(III) ions (Co 3+ ), nickel(II) ions (Ni 2+ ), and nickel(III) ions (Ni 3+ ). Complex ions of iron, cobalt, or nickel are also included in the iron group metal ions.

[0027] The function of the squalene derivative represented by formula (I) as a cation transporter is thought to originate from the crown ether moiety in formula (I). Therefore, when the crown ether moiety originates from 12-crown-4-ether (n=1), it is thought to exhibit high selectivity for lithium ions; when it originates from 15-crown-5-ether (n=2), it is thought to exhibit high selectivity for sodium ions; and when it originates from 18-crown-6-ether (n=3), it is thought to exhibit high selectivity for potassium ions.

[0028] The cation transport agent of the present invention can be used in membrane separation systems. The membrane separation system described above uses a separation membrane containing the cation transport agent of the present invention, i.e., a cation separation membrane.

[0029] The membrane material of the cation separation membrane is preferably one that can adequately hold the compound that acts as a cation transporter, i.e., the squalene derivative represented by formula (I) and the membrane solution that dissolves it, and has mechanical strength under membrane permeation conditions. There are no particular restrictions on materials that have such properties, but polymer materials, such as cellulose acetate, are preferred, and cellulose triacetate is more preferred. The membrane solution is selected appropriately depending on the compound used as the cation transporter, but it is preferable to use a solution that stably dissolves the cation transporter compound and has low dissolution into water from outside the membrane. For example, 2-nitrophenyl octyl ether (NPOE) is preferred. The solvent can be any solvent that can stably dissolve the compound that acts as a cation transporter, the film solution, and the film material, and can be easily removed after casting. For example, acetone, dimethylformamide, dimethyl sulfoxide, or chloroform are preferred, with chloroform being more preferred. Further details on the film manufacturing method are described, for example, in J.Membr.Sci., T.Shinbo, T.Yamaguchi, H.Yanagishita, K.Sakaki, D.Kitamoto, M.Sugiura, 84, 241-248 (1993), or in the Journal of the Textile Society of Japan, Toshio Shinbo, Toshiyuki Kanamori, Keiichi Ogasawara, Akihiro Yamazaki, Takashi Iwatsubo, Toshio Masuoka, Tomohiko Yamaguchi, 52, 105-109 (1996).

[0030] The amount of solvent used to dissolve the membrane material is preferably such that the membrane material accounts for 1 to 3% by mass of the solution after dissolution. While it is preferable to add as much membrane solution as possible relative to the membrane material, increasing the membrane solution content reduces the mechanical strength of the resulting membrane. Therefore, in practice, the amount of membrane solution is typically around 0.1 to 10 times the weight of the membrane material, and more generally, around 1 to 5 times. The amount of squalene derivative represented by formula (I) dissolved in the membrane material is appropriately determined based on the membrane material, membrane solvent, and the squalene derivative, but it is preferable to dissolve it in the membrane material in the highest possible proportion. Considering the practical limits of dissolution, 0.1 to 15% by mass is preferable, and 3 to 10% by mass is more preferable. It is preferable to stir the above mixed solution for as long as possible to obtain a homogeneous solution. Generally, 1 hour or more is preferable, and if possible, stirring for 24 hours or more is more preferable. The above solution can be spread to a certain thickness on a suitable smooth flat plate such as a glass plate and dried. The resulting film can then be carefully peeled off to obtain a cation separation membrane. The thickness of the separation membrane should preferably be as thin as possible, but as it becomes thinner, the mechanical strength of the resulting film decreases. Therefore, a thickness of 25 to 200 μm is preferred, and 25 to 100 μm or less is more preferred.

[0031] In the method of using the cation separation membrane of the present invention, i.e., the method of separating cations, preferably separated cations include alkali metal ions, alkaline earth metal ions, iron group metal ions, platinum group metal ions, and actinide metal ions. More preferably are alkali metal ions and alkaline earth metal ions, with alkali metal ions being particularly preferred. The above preferably separated cations (hereinafter also referred to as "target cations") may be dissolved in the separation solution as a single type or as multiple types simultaneously. They may also be dissolved together with other cations that are not intended for separation, in which case only the target cations can be selectively separated. The solvent used in the separation solution may be any solvent capable of dissolving the target cations, but an acidic aqueous solution is preferred, and an aqueous nitric acid solution is more preferred. The separation temperature is not particularly limited, but 5 to 50°C is preferred, and 35 to 45°C is more preferred. The initial concentration of the target cations contained in the separation solution is not particularly limited, but 1 to 10,000 ppm is preferred, and 10 to 1,000 ppm is more preferred.

[0032] [Anti-inflammatory drugs] The inventors of the present invention have discovered that the squalene derivative represented by the above formula (I) has excellent anti-inflammatory properties, and have come to invent the anti-inflammatory agent of the present invention.

[0033] The anti-inflammatory agent of the present invention contains a squalene derivative represented by formula (I) as an active ingredient. The squalene derivative represented by formula (I) is preferably one in which n=2 or 3.

[0034] The anti-inflammatory effect of the anti-inflammatory agent of the present invention can be evaluated, for example, as shown in the experimental example described later, by treating macrophages recovered from MafB region GFP knock-in mice, in which the coding region of the MafB (MAF bZIP transcription factor B) gene locus, a marker for M2 type macrophages, was replaced with the GFP (green fluorescent protein) gene, with the anti-inflammatory agent of the present invention and measuring the green fluorescence of GFP. Furthermore, the anti-inflammatory effect of the anti-inflammatory agent of the present invention can be confirmed, for example, by a cytotoxicity evaluation test using macrophage-like RAW cells. The MTT test, a cytotoxicity test, is a test that visualizes MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazole-3-ium bromide, yellow tetrazole) in living cells. In living cells, MTT is reduced to a purple formazan dye, but not in dead cells. Dimethyl sulfoxide, an acidic ethanol solution, or a dilute hydrochloric acid solution of sodium dodecyl sulfate (SDS) is added to solubilize the insoluble formazan dye, and the absorbance of the resulting colored solution between wavelengths of 500 and 600 nm is measured with a spectrophotometer to quantify living cells. The absorption maximum wavelength depends on the solvent used. In addition, a NO production measurement test is performed to confirm the inhibitory effect on the production of nitric oxide (NO), an inflammatory chemical substance induced by lipopolysaccharide (LPS). RAW cells are known to produce NO upon the addition of LPS, and this NO, either alone or in reaction with superoxide, produces DNA-damaging peroxynitrite, which damages cells and tissues and exacerbates inflammatory symptoms.

[0035] In the anti-inflammatory agent of the present invention, the squalene derivative represented by formula (I), which is the active ingredient, can be used alone or in combination of two or more types.

[0036] The anti-inflammatory agent of the present invention may be in the form of an anti-inflammatory composition comprising a squalene derivative represented by formula (I), which is an active ingredient, and other components.

[0037] In one embodiment, the anti-inflammatory composition of the present invention may contain a squalene derivative represented by the above formula (I) and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is not particularly limited and includes, for example, excipients, binders, disintegrants, lubricants, emulsifiers, thickeners, wetting agents, and solvents for injection. Furthermore, the anti-inflammatory composition of the present invention may further contain additives. The additives are not particularly limited and include, for example, preservatives, pH adjusters, stabilizers, UV absorbers, antioxidants, colorants, fragrances, and the like. For example, general raw materials listed in the Sixteenth Edition of the Japanese Pharmacopoeia can be used as the pharmaceutically acceptable carrier and the pharmaceutically acceptable additive.

[0038] Examples of dosage forms for anti-inflammatory compositions include orally administered forms such as tablets, coated tablets, pills, powders, granules, capsules, liquids, suspensions, and emulsions, as well as parenterally administered forms such as injections, suppositories, and topical preparations.

[0039] Examples of topical skin preparations include creams, lotions, toners, emulsions, foundations, packs, foams, ointments, patches, and aerosols.

[0040] The anti-inflammatory composition may be a treatment for alopecia, a cosmetic, or a food such as a supplement.

[0041] The content of the squalene derivative represented by formula (I) in the anti-inflammatory composition can range from 0.01 to 50% by mass, 0.01 to 30% by mass, 0.01 to 10% by mass, 0.01 to 5% by mass, or 0.01 to 1% by mass.

[0042] The method of administering anti-inflammatory agents or anti-inflammatory compositions is not particularly limited and should be determined appropriately according to the symptoms, weight, age, sex, etc. of the recipient. For example, tablets, coated tablets, pills, powders, granules, capsules, liquids, suspensions, emulsions, etc., are administered orally. Injectable preparations are administered intravenously, either alone or mixed with conventional infusion fluids such as glucose and amino acids, and may also be administered intra-arterial, intramuscular, intradermal, subcutaneous, or intraperitoneal as needed. Suppositories are administered rectally. Topical skin preparations are applied, patched, or sprayed onto the affected area.

[0043] The dosage of anti-inflammatory agents or anti-inflammatory compositions varies depending on the symptoms, weight, age, sex, etc., of the recipient and cannot be determined in general terms. However, for oral administration, for example, 0.01 to 500 mg / kg body weight of the active ingredient (squalene derivative represented by formula (I) above) per day should be administered. For injectable preparations, for example, 0.01 to 100 mg of the active ingredient per day should be administered. For suppositories, for example, 0.01 to 100 mg of the active ingredient per day should be administered. For topical skin preparations, for example, 0.01 to 500 mg of the active ingredient per day should be administered.

[0044] The anti-inflammatory agent of the present invention may be administered orally as a tablet or capsule, either as a pharmaceutical or nutritional supplement, or as a food or beverage, including Foods for Specified Health Uses and Foods with Function Claims. The anti-inflammatory agent of the present invention is useful for the prevention and treatment of inflammation. Because it has a macrophage activity inhibitory effect, it is effective against inflammation such as hay fever, allergic rhinitis, allergic conjunctivitis, and atopic dermatitis; inflammatory diseases such as delayed-type allergies, gastritis, and ulcerative colitis; arthritis such as rheumatoid arthritis and degenerative osteoarthritis; and other inflammatory diseases such as arteriosclerosis, endometriosis, acute respiratory distress syndrome, bronchitis, kidney transplant complications, acute myocardial infarction, diabetes mellitus, systemic lupus erythematosus, Crohn's disease, nephritis, hepatitis, pneumonia, IgA nephropathy, endotoxin shock, and sepsis due to infection.

[0045] When the anti-inflammatory agent of the present invention is used as a pharmaceutical or nutritional supplement, its shape is not particularly limited, and it can be formulated in any form, such as coated tablets, sugar-coated tablets, hard gelatin capsules, soft gelatin capsules, liquids, powders / granules, emulsions, or suspensions. There are no particular restrictions on the formulation carrier, and for example, one or more can be selected from among excipients, disintegrants, binders, surfactants, lubricants, coating agents, colorants, color fixatives, flavoring agents, antioxidants, preservatives, flavoring agents, acidulants, sweeteners, fortifiers, vitamins, leavening agents, thickeners, and flow enhancers, as long as they do not impair the various properties necessary for formulation and are appropriate to the dosage form of the final product.

[0046] Examples of the excipients include starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, and inorganic salts. Examples of the disintegrants include starch, hydroxypropyl starch, sodium carboxymethylcellulose, calcium carboxymethylcellulose, carboxymethylcellulose, and low-substituted hydroxypropylcellulose. Examples of the binders include starch, dextrin, acacia powder, gelatin, hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, and macrogol. Examples of the aforementioned surfactants include sodium lauryl sulfate, soy lecithin, sucrose fatty acid ester, and polysorbate 80. Examples of the lubricants include talc, waxes, hydrogenated vegetable oils, sucrose fatty acid esters, magnesium stearate, calcium stearate, aluminum stearate, and polyethylene glycol. Examples of the aforementioned fluidity enhancers include light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, and magnesium silicate.

[0047] The above-mentioned pharmaceuticals and nutritional supplements can be used for humans or other mammals. The dosage and intake amount for oral administration can be appropriately set according to the symptoms, sex, and age of the person using them. For example, an adult can take approximately 0.1 mg to 5 g of the active ingredient, a squalene derivative represented by formula (I), per day.

[0048] When the anti-inflammatory agent of the present invention is used as a food or beverage, it can be used not only as a food for specified health uses or a food with functional claims, but also as a general food or beverage. Examples of the aforementioned food and beverages include beverages (soft drinks, carbonated drinks, nutritional drinks, powdered drinks, fruit drinks, dairy drinks, jelly drinks, etc.), confectionery (cookies, cakes, gum, candy, tablets, gummies, steamed buns, yokan, pudding, jelly, ice cream, sherbet, etc.), processed seafood (fish sausage, kamaboko, chikuwa, hanpen, etc.), processed livestock products (hamburgers, ham, sausages, wieners, cheese, butter, yogurt, fresh cream, cheese, margarine, fermented milk, etc.), soups (powdered soups, liquid soups, etc.), staple foods (rice, noodles, bread, cereals, etc.), and seasonings (mayonnaise, shortening, dressings, sauces, dips, soy sauce, etc.). The food and beverages of the present invention may contain, for example, an amount of the squalene derivative represented by formula (I), which is the active ingredient, that can be ingested by an adult per day, ranging from approximately 0.1 mg to 5 g. [Examples]

[0049] The present invention will be described in more detail below with reference to experimental examples, but the present invention is not limited to the experimental examples described later.

[0050] [Experimental Example 1: Synthesis of Novel Squalene Derivatives (1)] From squalene of formula (1), 2,3-oxide squalene of formula (2) was synthesized by the method described above, and using this as a starting material, crown ether-added squalene (1,2) of formula (I) with n=2 was synthesized.

[0051] [ka]

[0052] <Synthesis of 15-crown-5-ether-added squalene (1,2)> 100 mg (0.23 mmol) of 2,3-epoxysqualene (compound of formula (2)) was dissolved in 8 mL of 2-propanol, and 2.8 g (11.2 mmol) of 2-hydroxymethyl-15-crown-5-ether was added. The mixture was heated at 80°C for 12 hours and allowed to cool. Water was added, and the mixture was extracted with ethyl acetate and dried over anhydrous magnesium sulfate. After concentrating the solvent under reduced pressure, the residue was purified using column chromatography (Wako Gel C-300) with a hexane-ethyl acetate mixed solvent to obtain 62 mg (40%) of 15-crown-5-ether-added squalene (2,2) as a colorless oil. The IR spectrum of this purified product was obtained. 1 H-NMR spectrum, 13 The 1C-NMR spectrum and HRMS (high-resolution mass spectrometry) are shown below.

[0053] Spectral data of 15-crown-5-ether-added squalene (1,2) IR(neat):3469,2913,2858,1731,1665,1445,1378,1357,1294,1247,1125,1090,984,934,840,738cm -1 . 1 H-NMR(400MHz,CDCl3)δ5.23-5.04(m,5H),3.88-3.56(m,19H),3.52-3.38(m,3H),2.38-2.23(m,1H),2.16 -1.90(m,18H),1.68(s,3H),1.66-1.54(brs,15H),1.49-1.35(m,2H),1.14-1.10(d,3H),1.09(s,3H)ppm. 13 C-NMR(100MHz,CDCl3)δ135.1(2C),135.0,134.9,131.2,124.5,124.4,124.2(3C),79.0,77.7,75.4,71.1,70.8(3C) ,70.6(4C),70.2,61.1,39.7(3C),37.0,29.8,28.2(2C),26.7(2C),26.6,25.7,21.5,19.8,17.6,16.0(3C),16.0ppm. HRMS(ESI,m / z,M +)m / z 676,5292,error 2.1ppm(calcd.for C 41 H 72 NaO7676.5278).

[0054] [ka]

[0055] [Experimental Example 2: Synthesis of Novel Squalene Derivatives (2)] From squalene of formula (1), 2,3-oxide squalene of formula (2) was synthesized by the method described above, and using this as a starting material, crown ether-added squalene (1,3) of formula (I) with n=3 was synthesized.

[0056] <Synthesis of 18-crown-6-ether-added squalene (1,3)> 100 mg (0.23 mmol) of 2,3-oxidesqualene (compound of formula (2)) was dissolved in 8 mL of 2-propanol, and 3.4 g (11.5 mmol) of 2-hydroxymethyl 18-crown-6-ether was added. The mixture was heated at 80°C for 6 hours and allowed to cool. Water was added, and the mixture was extracted with ethyl acetate and dried over anhydrous magnesium sulfate. After concentrating the solvent under reduced pressure, the residue was purified using column chromatography (Wako Gel C-300) with a hexane-ethyl acetate mixed solvent to obtain 72 mg (47%) of 18-crown-6-ether-added squalene (2,2) as a colorless oil. The IR spectrum of this purified product was obtained. 1 H-NMR spectrum, 13 The 1C-NMR spectrum and HRMS (high-resolution mass spectrometry) are shown below.

[0057] Spectral data of 18-crown-6-ether-added squalene (1,3) IR(neat)3480,2915,2861,1732,1664,1446,1378,1354,1295,1247,1112,988,943,842,737cm -1 . 1H-NMR(400MHz,CDCl3)δ5.21-5.05(m,5H),3.88-3.59(m,23H),3.52-3.41(m,3H),2.24-2.35(m,1H),2.15 -1.91(m,18H),1.68(s,3H),1.66-1.55(brs,15H),1.48-1.36(m,2H),1.14-1.10(d,3H),1.09(s,3H)ppm. 13 C-NMR(100MHz,CDCl3)δ135.1(2C),135.0,134.9,131.2,124.5,124.4,124.3(3C),78.7,77.7,75.2,71.3,70.8(4C) ,70.6(5C),69.9,61.1,39.7(3C),37.0,29.8,28.2(2C),26.7(2C),26.6,25.7,21.5,19.8,17.6,16.0(3C),16.0ppm. HRMS(ESI,m / z,M + )m / z 720.5541,error 0.1ppm(calcd,for C 34 H 60 NaO4720.5540).

[0058] [Experimental Example 3: Evaluation of Cation Retention Function (1)] The association constants of crown ether-added squalene (1,2) with respect to alkali metal ions were determined by the method described in P. Thordarson, “Determining association constants from titration experiments in supramolecular chemistry”, Chem. Soc. Rev., 40(3), 2011, 1305-1323.

[0059] The association constant (K) of crown ether-added squalene (1,2) with respect to alkali metal ions a )(vs Anion: SCN - The solvent (CH3CN) is shown in Table 1.

[0060] [Table 1]

[0061] Crown ether-added squalene (1,2) and alkali metal ions exhibited a 1:1 association characteristic. Crown ether-added squalene (1,2) exhibited selectivity for sodium ions and possessed the largest association constant.

[0062] The chemical shift of the crown ether moiety of crown ether-added squalene (1,2) at around 3.5 ppm shifted to a low magnetic field upon association with alkali metal ions. This associated state remained stable for more than 6 months. For example, the case of crown ether-added squalene (1,2) alone (top panel) and the case of the association state of crown ether-added squalene (1,2) with sodium ions (bottom panel) 1 The 1H-NMR spectrum is shown in Figure 1 [solvent: CD3CN, concentration of crown ether adducted squalene (1,2) = 1.0 × 10⁻¹⁰]. -3 M,Na + Concentration=1.0×10 -2 M].

[0063] [Experimental Example 4: Evaluation of Cation Retention Function (2)] The association constants of crown ether-added squalene (1,3) with alkali metal ions were determined in the same manner as in Experimental Example 3.

[0064] The association constant (K) of crown ether-added squalene (1,3) with alkali metal ions. a )(vs Anion: SCN - The solvent (CH3CN) is shown in Table 2.

[0065] [Table 2]

[0066] Crown ether-added squalene (1,3) and alkali metal ions exhibited a 1:1 association characteristic. Crown ether-added squalene (1,3) exhibited selectivity for potassium ions and possessed the largest association constant.

[0067] The chemical shift of the crown ether moiety of crown ether-added squalene (1,3) at around 3.5 ppm shifted to a low magnetic field upon association with alkali metal ions. This associated state remained stable for more than 6 months. For example, the case of crown ether-added squalene (1,3) alone (top panel) and the case of the association state of crown ether-added squalene (1,3) with potassium ions. 1 The 1H-NMR spectrum is shown in Figure 2 [solvent: CD3CN, concentration of crown ether adducted squalene (1,3) = 1.0 × 10⁻¹⁰]. -3 M,K + Concentration=1.0×10 -2 M].

[0068] [Experimental Example 5: Evaluation of Anti-inflammatory Effects] The anti-inflammatory effect of the squalene derivative represented by formula (I) was evaluated by treating macrophages recovered from MafB region GFP knock-in mice, in which the coding region of the MafB (MAF bZIP transcription factor B) gene locus, a marker for M2 type macrophages, was replaced with the GFP (green fluorescent protein) gene, with the anti-inflammatory agent of the present invention and measuring the green fluorescence of GFP.

[0069] <Experimental Procedure> (1) Inflammation was induced by injecting thioglycolate medium (5%, 2 mL) into the peritoneal cavity of MafB region GFP knock-in mice (obtained from the Animal Resource Center of the University of Tsukuba). (2) Three days after administration of thioglycolate medium, peritoneal fluid containing macrophages was collected from the peritoneal cavity of mice. (3) The collected peritoneal fluid was centrifuged at 1000 × g for 3 minutes to collect macrophages. (4) Place the collected macrophages in a microplate containing culture medium in a 1 × 10⁶ 5Seeds were seeded to a concentration of cells / mL. The culture medium consisted of 300 mL DMEM (high glucose), 50 mL FBS (fetal bovine serum), and 5 mL PS (penicillin streptomycin). (5) After 24 hours of incubation, the crown ether-added squalene (1,2) synthesized in Experimental Example 1 was added to each well of the microplate to final concentrations of 0 μg / mL (control), 10 μg / mL, 25 μg / mL, 50 μg / mL, 75 μg / mL, and 100 μg / mL. (6) After 24 hours of culture, GFP expression was analyzed. GFP expression was analyzed using image analysis to measure the number of GFP-expressing cells, the area of ​​GFP expression, and the intensity of GFP expression. The results of the GFP expression analysis are shown in Figure 3 (number of GFP-expressing cells), Figure 4 (GFP expression area), and Figure 5 (GFP expression intensity).

[0070] Treatment of macrophages with the novel squalene derivative of the present invention resulted in a significant increase in the number of GFP-expressing cells, a significant increase in the GFP-expressing area, and a significant increase in GFP expression intensity, suggesting that the novel squalene derivative of the present invention has anti-inflammatory effects. [Industrial applicability]

[0071] The cation transporter of the present invention is particularly useful in membrane separation systems because, within a system where a small amount of transporter in the membrane comes into contact with the supplying aqueous phase, takes in cations into the membrane, solubilizes them, moves through the membrane, and releases them at the interface with the receiving aqueous phase, a large amount of cations can be continuously separated and concentrated using only a small amount of transporter. Furthermore, inflammatory responses in living organisms are not only biological defense reactions that occur when harmful stimuli such as the invasion of foreign substances, infection, trauma, burns, or allergens act on the body, but inflammation is also thought to be the underlying pathogenesis of many diseases. The novel squalene derivative of the present invention exhibits anti-inflammatory effects at lower concentrations, and is therefore expected to have anti-inflammatory effects and symptom improvement effects as an easily ingestible anti-inflammatory agent.

Claims

1. A squalene derivative represented by the following formula (I). 【Chemistry 1】 However, in equation (I), n is an integer between 1 and 5, inclusive.

2. The squalene derivative according to claim 1, wherein n in formula (I) is 2 or 3.

3. A cation transport agent comprising the squalene derivative described in claim 1 or 2 as an active ingredient.

4. A cation transport agent according to claim 3, used in a cation membrane separation system.

5. A cation transport agent according to claim 3 for transporting at least one metal ion selected from the group consisting of alkali metal ions, alkaline earth metal ions, iron group metal ions, platinum group metal ions, and actinide metal ions.

6. An anti-inflammatory agent comprising the squalene derivative described in claim 1 or 2 as an active ingredient.

7. Food and beverages comprising the squalene derivative according to claim 1 or 2.

8. A pharmaceutical product comprising the squalene derivative described in claim 1 or 2.

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