JP2020114871A - Solution containing stabilized mycosporin-like amino acid, and method for producing the same - Google Patents

Abstract

To provide solution containing stabilized mycosporin-like amino acid (MAA).SOLUTION: A method for inhibiting decomposition of MAA in solution, wherein (a) MAA, and (b) one or more substances selected from the group consisting of α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, methionine, serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, parahydroxy sodium benzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, and ergothioneine are mixed with the solution.SELECTED DRAWING: None

Description

The present invention relates to a solution containing a stabilized mycosporine-like amino acid and a method for producing the same. The invention also relates to a method of stabilizing a solution containing a mycosporine-like amino acid.

Ultraviolet radiation (UV) is known to induce acute skin reactions such as erythema, skin aging and skin cancer. The ultraviolet rays contained in sunlight are classified into three types, UV-A (320 nm-400 nm), UV-B (280 nm-320 nm) and UV-C (200-280 nm), depending on the wavelength. Of these, UV-A and UV-B have an effect on the living body, and UV-C does not pose a problem because it usually cannot pass through the atmosphere.

It is known that UV-B is the main cause of sunburn outdoors and has a relatively large energy as compared with UV-A. When absorbed into the skin layer, it reaches the stratum corneum and epidermis, causing acute skin pigmentation such as spots and freckles. It is also known to cause immunosuppression, which is involved in aging and development of skin cancer.

It is known that UV-A has lower energy but longer wavelength than UV-B, penetrates deeper into the skin than UV-B, and reaches the dermis. As a result, not only acute skin pigmentation such as spots and freckles, but also a decrease in elasticity of the dermis (actinic elastic fibrosis) is caused, and early skin aging phenomena such as wrinkles and sagging are caused. More recently, it has been found that UV-A also causes immunosuppression and is involved in the development of precancerous skin lesions and skin cancer.

The amount of UV-B varies depending on the season, weather, latitude, etc., whereas UV-A reaches a certain amount throughout the year. Therefore, it is important to protect the skin from UV-A as well as UV-B.

Current UV protection agents can be classified into UV absorbers and UV scatterers. The ultraviolet absorber converts ultraviolet energy into heat energy and emits it, and examples thereof include synthetic organic compounds such as 4-tert-butyl-4′-methoxydibenzoylmethane. The ultraviolet scattering agent contains inorganic particles such as titanium oxide (TiO ₂ ) and zinc oxide (ZnO), and when applied to the skin, the inorganic particles present on the skin surface function as a barrier that reflects ultraviolet rays.

UV absorbers are (1) easily decomposed by light and have poor stability, (2) they cause itching and allergies by promoting molecular excitation and melanin production, and (3) because they are chemically synthesized substances. However, there is a problem that the image given to the user is bad. UV scattering agents (1) when applied to the skin tend to become white and give the skin a heavy feel, (2) cause the production of active oxygen, (3) block pores in the skin, and function as an exocrine gland. There is a problem that there is a concern that this will be hindered. Due to these problems, expectations for safe UV-absorbing substances of natural origin are increasing.

Mycosporine-like amino acid (hereinafter referred to as “MAA”) is a natural UV absorbing substance that is widely known to exist in aquatic organisms such as coral, red algae, viscera and microalgae. is there.

So far, attempts have been made to produce MAA by chemical synthesis or a light irradiation method using cyanobacteria (Patent Document 1 and Non-Patent Document 1). Furthermore, it has also been attempted to extract MAA from natural products such as Nori, algae and shellfish (Patent Document 2 and Non-Patent Documents 2 to 4). In addition, the present inventors have previously developed a method for producing a mycosporine-like amino acid using a microorganism (Patent Document 3).

International Publication No. 02/39974 Special table 2013-518871 gazette International Publication No. 2015/174427

World J Microbiol Biotechnol (2008) 24:3111-3115. Marine Biology 108, 157-166 (1991). Photomedicine and Photobiology (2002), 24, 39-42. Tetrahedron Letters 1979, 3181-3182.

It has been pointed out that MAA is colored in a solution state. However, a method for preventing this coloring has not been found so far. In addition, the present inventors have found a problem that MAA is decomposed in a solution in the process of producing and purifying MAA, and a problem that the UV absorption ability is lowered when the MAA solution is applied to the skin or the like. It was The problem to be solved by the present invention includes solving these problems.

The present inventors say that the addition of MAA-containing solution results in (1) coloring of the MAA solution, (2) decomposition of MAA in the solution, and (3) reduction of UV absorption ability of MAA in the thin film solution. Have found a substance that solves the problem of stability of MAA solution, and completed the present invention.

That is, the present invention provides the following:
[1] A solution containing stabilized mycosporine-like amino acid,
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. A solution comprising one or more substances selected from the group and (c) a solvent;
[2] A solution containing a stabilized mycosporine-like amino acid,
(A) mycosporine-like amino acid, and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A solution containing a solvent;
[3] A solution containing a stabilized mycosporine-like amino acid,
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. One or more substances selected from the group, and (c) methanol, ethanol, 1-propanol, 2-propanol, glycerol, 1,3-butanediol, 1,4-butanediol, propylene glycol, ethylene glycol, diethylene glycol, A solution comprising a solvent containing one or more substances selected from the group consisting of triethylene glycol, tetraethylene glycol, N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO) and pentaethylene glycol;
[4] The solution according to any one of [1] to [3] used for an agent for skin.
[5] The solution according to any one of [1] to [3] used for medicines;
[6] The solution according to any one of [1] to [3] used in cosmetics;
[7] A method for producing a solution containing a stabilized mycosporine-like amino acid, comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. A method comprising mixing one or more substances selected from the group, and (c) a solvent;
[8] A method for producing a solution containing a stabilized mycosporine-like amino acid, comprising:
(A) mycosporine-like amino acid, and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A method comprising: mixing a solvent; and [9] a method of producing a solution containing a stabilized mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. One or more substances selected from the group, and (c) methanol, ethanol, 1-propanol, 2-propanol, glycerol, 1,3-butanediol, 1,4-butanediol, propylene glycol, ethylene glycol, diethylene glycol, A method comprising mixing a solvent comprising one or more substances selected from the group consisting of triethylene glycol, tetraethylene glycol, N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO) and pentaethylene glycol. ..

According to the present invention, a solution containing stabilized MAA can be provided.

In one embodiment, the invention provides a solution containing a stabilized mycosporine-like amino acid,
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. One or more substances selected from the group,
And (c) a solvent is provided.

By adding the substance described in (b) above, (1) coloring of the MAA solution and/or (2) decomposition of MAA in the solution can be prevented.

In the present embodiment, as the “solvent” described in (c) above, any solvent that dissolves mycosporine-like amino acid can be used. Examples of such a solvent include, but are not limited to, water or a mixed liquid of water and another substance.

In another embodiment, a solution containing a stabilized mycosporine-like amino acid comprising:
(A) mycosporine-like amino acid, and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A solution is provided that includes a solvent.

By adding the substance described in the above (c), it is possible to prevent (3) the decrease in the UV absorption ability of MAA in the thin film solution.

In this embodiment, the “solvent” described in (c) above may or may not contain water.

In another embodiment, a solution containing a stabilized mycosporine-like amino acid comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. One or more substances selected from the group, and (c) methanol, ethanol, 1-propanol, 2-propanol, glycerol, 1,3-butanediol, 1,4-butanediol, propylene glycol, ethylene glycol, diethylene glycol, A solution is provided that includes a solvent that includes one or more substances selected from the group consisting of triethylene glycol, tetraethylene glycol, N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO) and pentaethylene glycol.

By adding the substance described in (b) above and the substance described in (c) above, (1) coloring of the MAA solution and/or (2) decomposition of MAA in the solution is prevented, And (3) the decrease in the UV absorption ability of MAA in the thin film solution can be prevented.

In this embodiment, the “solvent” described in (c) above may or may not contain water.

In the embodiment described above, the amount of the substance described in (a) added is 0.0001 to 10% by weight, preferably 0.001 to 1% by weight, and more preferably 0.005 to 0% by weight based on the total amount of the solution. It is 0.5% by weight.

In the above-described embodiment, the amount of the substance described in (b) above is 0.00001 to 20% by weight, preferably 0.0001 to 10% by weight, and more preferably 0.001 to 10% by weight based on the total amount of the solution. 5% by weight, particularly preferably 0.01 to 2% by weight.

In the embodiment described above, the substance described in (c) above is added in a proportion of, for example, 50% by weight or more, 60% by weight or more, 70% by weight or more, or 80% by weight or more with respect to the total amount of the solvent. Preferably, the substance described in (c) is added in a proportion of 90% by weight or more based on the total amount of the solvent. In one embodiment, such a solvent is a mixture of 50 wt% or more of the substance described in (c) and 50 wt% or less of water. For example, a mixture of 60% by weight or more of the substance described in (c) and 40% by weight or less of water, a mixture of 70% by weight or more of the substance of (c) and 30% by weight or less of water, or 80 It is a mixture consisting of not less than 20% by weight of water and not less than 20% by weight of the substance described in (c). Preferred is a mixture consisting of 90% by weight or more of the substance (c) and 10% by weight or less of water.

As used herein, "stabilizing" means not only stabilizing the mycosporine-like amino acid but also reducing the change in the state of the solution containing the mycosporine-like amino acid.

As used herein, “mycosporine-like amino acid” and “MAA” are generic terms for compounds in which an amino acid, amine, ammonia or amino alcohol is bonded to a cyclohexenone or cyclohexene imine skeleton which may have a substituent. is there.

［式（Ｉ）中、Ｑ^１は酸素原子、アミノ酸、アミン、アンモニアまたはアミノアルコールであり、Ｑ^１がアミノ酸、アミン、アンモニアまたはアミノアルコールである場合、該Ｑ^１は窒素原子を介して環に結合しているものであり；Ｑ^２はアミノ酸、アミン、アンモニアまたはアミノアルコールであり、該Ｑ^２は窒素原子を介して環に結合しているものである］
で示される化合物である。 Specifically, the MAA used in the present invention has the formula (I):

[In the formula (I), Q ¹ is an oxygen atom, an amino acid, an amine, ammonia or an aminoalcohol, and when Q ¹ is an amino acid, an amine, ammonia or an aminoalcohol, the Q ¹ is bonded to the ring via a nitrogen atom. Q ² is an amino acid, an amine, ammonia or an amino alcohol, and Q ² is bound to the ring via a nitrogen atom]
Is a compound represented by.

In one embodiment, Q ¹ is selected from the group consisting of an oxygen atom, glycine, alanine, valine, serine, threonine, leucine, isoleucine, asparagine, aspartic acid, glutamine, and glutamic acid, and Q ² is valine, asparagine, It is selected from the group consisting of aspartic acid, threonine, leucine, isoleucine, alanine, glycine, serine, glutamine, and glutamic acid.

In other embodiments, the combination of Q ¹ and Q ^{2 is} a combination of any one of the tables below.

In another embodiment, the MAA is cynoline (hereinafter, formula (II)), porphyra-334 (hereinafter, formula (III)), asterina-330 (hereinafter, formula (IV)), paritene (hereinafter, formula (V )), palytin (hereinafter, formula (VI)), mycosporine-glycine (hereinafter, formula (VII)), mycosporine-glycine: valine (hereinafter, formula (VIII)), mycosporine serinol (hereinafter, formula (IX)). ) Or mycosporin-glycine-alanine (hereinafter, formula (X)).

As used herein, MAA includes compounds represented by the above formulas (I) to (X), as well as pharmaceutically acceptable salts, prodrugs, oxides, ethers, esters, glycosides, or conjugates thereof. The body is also included. Alternatively, MAA used in the present invention may be a combination of the compounds represented by the above formulas (I) to (X).

In the present specification, "pharmaceutically acceptable salt" includes inorganic acids such as sulfuric acid, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, perchloric acid and carbonic acid. Salt, or
Acetic acid, benzoic acid, oxalic acid, lactic acid, malic acid, tartaric acid, succinic acid, fumaric acid, maleic acid, citric acid, malonic acid, mandelic acid, gluconic acid, galactaric acid, glucoheptonic acid, glycolic acid, aspartic acid, glutamic acid, Salts with organic acids such as trifluoroacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, camphorsulfonic acid, or naphthalene-2-sulfonic acid, lithium ion, sodium ion, potassium ion, Salts with one or more metal ions such as calcium ion, magnesium ion, zinc ion, aluminum ion and silver ion, ammonia, arginine, lysine, piperazine, N-methylpiperazine, 1-(2-hydroxyethyl)piperazine, 1,3-diaminopropane, choline, diethylamine, trimethylamine, triethylamine, dicyclohexylamine, 4-phenylcyclohexylamine, ethanolamine, diethanolamine, 2-amino-1-propanol, 1,3-diamino-2-propanol, benzathine, N -Salts with amines such as methylmorpholine, prolinol, 2-piperidinemethanol are included. These salts can be manufactured according to a conventionally known method.

“Prodrug” means a derivative of any of the above compounds which, when administered to a subject, is capable of directly or indirectly obtaining the above compound or an active metabolite or residue thereof. Prodrugs may have the following advantages: improved bioavailability of the compound; reduced side effects and/or toxicity of the compound; enhanced delivery of the compound to the biological area; half-life of the compounds described above. Of the compound; stabilization of the compound; improvement of the taste and odor of the compound. Prodrugs include, for example, the ester forms of the compounds described above. Examples of ester prodrugs include formate, acetate, propionate, butyrate, acrylate, farnesylate and ethylsuccinate derivatives. For a general description of prodrugs, see T.W. Higuchi and V.I. Stella, Pro-drugs as Novel Drug Delivery Systems, Vol. 14 of the A. C. S. Symposium Series and Edward B. Roche, ed. , Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, all of which are incorporated herein by reference.

“Compound” further includes isomers such as structural isomers (eg, chain isomers, positional isomers, and geometric isomers), stereoisomers (eg, rotamers), and tautomers of the compounds. In this case, the "compound" includes any one isomer and a mixture of isomers. Each of these isomers can be obtained as a single product by a conventionally known synthesis method or separation method (concentration, solvent extraction, column chromatography, recrystallization, etc.). Furthermore, "compound" also includes all possible resonance forms. Furthermore, "compound" also includes all possible labeled compounds.

Thus, the amino acid residues included in formula (I) include structural isomers of any amino acid. For example, when the amino acid residue contained in formula (I) is alanine, the amino acid residue may be α-alanine or β-alanine. When the amino acid residue contained in formula (I) is lysine, the amino acid residue may be α-lysine or β-lysine. When the amino acid residue contained in formula (I) is leucine or isoleucine, the amino acid residue may be norleucine. When the amino acid residue contained in formula (I) is valine, the amino acid residue may be norvaline or isovaline.

Further, when both Q ¹ and Q ^{2 in} the formula (I) are amino acids, amines, ammonia or amino alcohols, these may be replaced with each other.

"Stereoisomer" means molecules that differ in the chirality of one or more stereocenters. Stereoisomers include enantiomers, diastereomers and mixtures thereof. The stereoisomers also include "rotational isomers". "Rotamers" mean molecules with different conformations that have been made separable by virtue of the increased rotation barrier around single bonds, due to large steric hindrance and the like.

Thus, the amino acid residues included in formula (I) include stereoisomers of any amino acid. That is, the amino acid residue contained in formula (I) may be an L-amino acid residue or a D-amino acid residue. When the amino acid residue contained in formula (I) is any amino acid having two or more asymmetric carbon atoms, the amino acid residue contained in formula (I) is a diastereomer of the amino acid. Also includes. For example, when the amino acid residue contained in formula (I) is isoleucine, the amino acid residue may be L-isoleucine or D-isoleucine. The amino acid residue may also be L-alloisoleucine or D-alloisoleucine. When the amino acid residue contained in formula (I) is threonine, the amino acid residue may be L-threonine or D-threonine. The amino acid residue may also be L-allosereonine or D-allosereonine.

When Q ¹ and Q ² contained in the formula (I) are residues different from each other, the carbon atom marked with an asterisk (*) in the following formula (I′) becomes an asymmetric carbon atom. In this case, the configuration may be either S configuration or R configuration.

The “tautomer” refers to a compound that can reach an equilibrium state in which both isomers coexist with each other at a high rate of isomerization in which the isomers convert each other. Tautomerism includes, for example, proton tautomerism due to 1,3-rearrangement of protons.

The MAA used in the present invention may include not only a MAA that has already been structurally determined but also a MAA having a novel structure. Further, the MAA used in the present invention may be a naturally occurring MAA or an artificially produced MAA.

As a method for identifying MAA, a conventionally known method can be used. For example, high performance liquid chromatography-time of flight mass spectrometry (HPLC-TOFMS) may be used. Alternatively, MAA may be identified by combining high resolution mass spectrometry (HR-MS) and nuclear magnetic resonance (NMR). Alternatively, HPLC may be used to identify MAA from retention time and UV spectrum results. Alternatively, MAA may be identified by measuring the UV absorption spectrum and exact mass using HPLC equipped with a photodiode array detector and an HR-MS detector.

In another embodiment, the solutions of the invention described above are used in cosmetics, quasi drugs, or pharmaceuticals. Examples of cosmetics, quasi drugs, or pharmaceuticals include, but are not limited to, dermatological agents. The “dermatological agent” may be applied externally. Alternatively, it may be used by other methods such as oral administration, injection and infusion.

Examples of "cosmetics" and "quasi drugs" include, but are not limited to, those for skin, hair, eyes, and nails. For example, lotion, emulsion, gel, beauty essence, cream, sunscreen, sunscreen spray, pack, mask, foundation, powder, manicure (base coat, color polish and top coat, etc.), bath agent, antiperspirant, vitamin Agents, body lotions, shampoos, rinses, hair treatments, hair conditioners, hair colors, hair styling agents, hair tonics, and hair restorers, but are not limited thereto.

Examples of "pharmaceutical products" include, but are not limited to, those for skin, hair, eyes, and nails. For example, a drug for preventing or treating an acute skin reaction, a drug for preventing or improving skin aging, a drug for preventing or treating skin cancer, a drug for growing hair, a drug for preventing or treating eye diseases. Medications and medicinal products for preventing or treating ringworm include, but are not limited to.

The forms of "cosmetics", "quasi drugs", or "pharmaceuticals" include liquids, creams, lotions, pastes, ointments, emulsions (oil-in-water emulsion, water-in-oil emulsion, multiple emulsion, microemulsion, PET). -Emulsions, Pickering emulsions), gels (hydrogels, alcohol gels), suspensions, foams, sprays, tablets, powders, eye drops, eye ointments or patches and the like, but are not limited thereto.

“Cosmetic”, “quasi drug”, or “pharmaceutical product” means an auxiliary agent such as a moisturizer, a surfactant, a pigment, a fragrance, an antiseptic, a bactericide, and an antioxidant, which is usually used for these. You may include an additive.

In another embodiment, the above-mentioned solution of the present invention is used for an ultraviolet absorbing composition such as a coating composition and other coating agents. The coating composition and other coating agents can be prepared by using a method known to those skilled in the art, such as a spray method, a dipping method, a roller coating method, a flow coater method, a flow coating method, etc., for a substrate (metal, plastic, wood, ceramic, It is applied or coated on glass, fiber, paper, etc.). The composition for absorbing ultraviolet rays can be applied to industrial applications, medical devices, and the like. For example, the ultraviolet absorbing composition may be applied to contact lenses and spectacle lenses.

In another embodiment, the invention provides a method of making a solution containing a stabilized mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. There is provided a method comprising mixing one or more substances selected from the group and (c) a solvent.

The above method can prevent (1) coloring of the MAA solution and/or (2) decomposition of the MAA in the solution.

In the present embodiment, as the “solvent” described in (c) above, any solvent that dissolves mycosporine-like amino acid can be used. Examples of such a solvent include, but are not limited to, water or a mixed liquid of water and another substance.

In another embodiment, the invention provides a method of making a solution containing a stabilized mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid, and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A method is provided that includes mixing a solvent.

By the above method, (3) the decrease in the UV absorption ability of MAA in the thin film solution can be prevented.

In this embodiment, the “solvent” described in (c) above may or may not contain water.

In another embodiment, the invention provides a method of making a solution containing a stabilized mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. One or more substances selected from the group, and (c) methanol, ethanol, 1-propanol, 2-propanol, glycerol, 1,3-butanediol, 1,4-butanediol, propylene glycol, ethylene glycol, diethylene glycol, A method comprising mixing a solvent comprising one or more substances selected from the group consisting of triethylene glycol, tetraethylene glycol, N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO) and pentaethylene glycol. I will provide a.

The above method can prevent (1) coloring of the MAA solution and/or (2) decomposition of the MAA in the solution, and (3) reduction of the UV absorption ability of the MAA in the thin film solution.

In the above-mentioned production method of the present invention, the substance described in (b) above is added to the solution at the above-mentioned concentration. Further, the substance described in (c) above is added to the solution at the above-mentioned ratio.

The above (a) to (c) may be mixed at the same time or individually. Further, the above (a) to (c) may be mixed in any order.

In yet another embodiment, the invention provides a method of stabilizing a solution containing a mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. There is provided a method comprising mixing one or more substances selected from the group and (c) a solvent.

The above method can prevent (1) coloring of the MAA solution and/or (2) decomposition of the MAA in the solution. Thereby, the solution containing MAA can be stabilized.

In the present embodiment, as the “solvent” described in (c) above, any solvent that dissolves mycosporine-like amino acid can be used. Examples of such a solvent include, but are not limited to, water or a mixed liquid of water and another substance.

In yet another embodiment, the invention provides a method of stabilizing a solution containing a mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid, and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A method is provided that includes mixing a solvent.

By the above method, (3) the decrease in the UV absorption ability of MAA in the thin film solution can be prevented. Thereby, the solution containing MAA can be stabilized.

In this embodiment, the “solvent” described in (c) above may or may not contain water.

In yet another embodiment, the invention provides a method of stabilizing a solution containing a mycosporine-like amino acid, the method comprising:
(A) mycosporine-like amino acid,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, Consists of serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate. One or more substances selected from the group, and (c) methanol, ethanol, 1-propanol, 2-propanol, glycerol, 1,3-butanediol, 1,4-butanediol, propylene glycol, ethylene glycol, diethylene glycol, A method comprising mixing a solvent comprising one or more substances selected from the group consisting of triethylene glycol, tetraethylene glycol, N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO) and pentaethylene glycol. I will provide a.

The above method can prevent (1) coloring of the MAA solution and/or (2) decomposition of the MAA in the solution, and (3) reduction of the UV absorption ability of the MAA in the thin film solution. Thereby, the solution containing MAA can be stabilized.

In this embodiment, the “solvent” described in (c) above may or may not contain water.

In the stabilization method of the present invention described above, the substance described in (b) above is added to the solution at the concentration described above. Further, the substance described in (c) above is added to the solution at the above-mentioned ratio.

The above (a) to (c) may be mixed at the same time or individually. Further, the above (a) to (c) may be mixed in any order.

Hereinafter, the present invention will be described specifically and in detail with reference to examples, but the examples are used for illustrating the present invention and are not intended to limit the present invention.

(Example 1)
Cinoline 9 mg and each of the substances listed in the table below were dissolved in 9 ml of 0.1 M phosphate buffer (sodium dihydrogen phosphate + disodium hydrogen phosphate; pH 7), and the mixture was shielded from light at 50°C for 2 weeks. Heated. The absorbance at 400 nm before and after the reaction was measured to evaluate the effect of each substance for suppressing the coloration of the shinorin solution. Compared with the negative control solution containing only shinorin, the solutions in which the substances shown in the table were present showed a small increase in absorbance at the start of the reaction and after 2 weeks.

(Comparative Example 1)
Sinoline 9 mg and niacinamide, ascorbic acid, L-ascorbic acid 2-glucoside, α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, coumarin, gallic acid, EDTA, sodium gluconate, sodium tartrate, serine, glucose , Allose, β-cyclodextrin, and glucosamine hydrochloride (18 mg) each were dissolved in 9 ml of 0.1 M phosphate buffer (sodium dihydrogen phosphate + disodium hydrogen phosphate; pH 7) and shielded from light at 50°C. And warmed for 2 weeks. The absorbance at 400 nm before and after the reaction was measured. Compared to the negative control solution containing only shinorin, the solution containing these substances showed a large increase in absorbance at the start of the reaction and after 2 weeks.

＜ＨＰＬＣ分析条件＞
カラム： ＹＭＣ−Ｐａｃｋ ＯＤＳ−ＡＱ Ｓ−５μＭ、１２ｎｍ ２５０×４．６ｍｍＩ．Ｄ．
流速： ０．５ｍｌ／分
温度： ３０℃
検出： ＰＤＡ（３３４ｎｍ）
溶離液：０．１％ギ酸水溶液
シノリンの保持時間： ８分付近 (Example 2)
Cinoline 9 mg and each of the substances listed in the table below were dissolved in 9 ml of 0.1 M phosphate buffer (sodium dihydrogen phosphate + disodium hydrogen phosphate; pH 7), and the mixture was shielded from light at 50°C for 2 weeks. Heated. The cynoline concentration before and after the reaction was measured under the HPLC conditions described below. In the table, in the "degradation inhibition" column, ◎ indicates a very high residual cynoline rate, and ○ indicates a high residual cynoline rate, as compared to the negative control solution containing only cynoline. ing.

<HPLC analysis conditions>
Column: YMC-Pack ODS-AQ S-5 μM, 12 nm 250×4.6 mm I.D. D.
Flow rate: 0.5 ml/min Temperature: 30°C
Detection: PDA (334nm)
Eluent: 0.1% formic acid aqueous solution Retention time of shinorin: About 8 minutes

In addition, the same experiment was performed using niacinamide, L-ascorbic acid 2-glucoside, coumarin, sodium gluconate, sodium tartrate, arginine, cysteine, and tocopherol acetate (18 mg), respectively. However, no improvement effect of 5% or more was observed on the cynoline residual rate.

(Comparative example 2)
Dissolve 9 mg of cynoline and 18 mg of ascorbic acid, which is an isomer of erythorbic acid, in 9 ml of 0.1 M phosphate buffer (sodium dihydrogen phosphate + disodium hydrogen phosphate; pH 7) and add at 50°C for 2 weeks under light shielding. Warmed. The cynoline concentration before and after the reaction was measured under the HPLC conditions described in Example 2. As a result, contrary to the result when erythorbic acid was used, the residual ratio of cynoline was lower than the residual ratio of the shinorin solution alone.

＊商品名「トルナーレ（登録商標）」、株式会社林原販売 (Example 3)
Cinoline 9 mg and a combination of two or more substances listed in the table below were each dissolved in 9 ml of 0.1 M phosphate buffer (sodium dihydrogen phosphate + disodium hydrogen phosphate; pH 7) and shielded from light at 50°C. And warmed for 2 weeks. The shinorin concentration before and after the reaction was measured under the HPLC conditions described in Reference Example 2 and the absorbance at 400 nm was measured. In the table, in the "coloration suppression" column, the ones with a small increase in absorbance at the start of the reaction and two weeks after the reaction are indicated by ◯, and the ones with a large increase are indicated by x, as compared with the negative control solution containing only shinorin. .. In the “degradation inhibition” column, “⊚” indicates a very high cynoline residual rate, and “∘” indicates a high cynoline residual rate, as compared with the negative control solution containing only cynoline.

*Product name "Tornare (registered trademark)", Hayashibara Sales Co., Ltd.

(Example 4)
The quartz plate was attached to UV-STAR MICROPLATE, 96 Well. 16 cm to 50 mg of an aqueous solution shown in the following table containing 0.5% cynoline was thinly applied to 4 cm×4 cm (16 cm ² ) including nine points of Well numbers B9-B11, C9-C11, and D9-D11. The plate was left at room temperature for 24 hours. The average value of the absorbance at 334 nm was measured at 9 points of B9-B11, C9-C11, and D9-D11 immediately after coating and after 24 hours. In the table, in the "UV absorption effect persistence" column, 80% or more after 24 hours compared with the absorbance immediately after coating is indicated by ◯, and 50% or more and less than 80% is indicated by Δ. The 90% glycerol aqueous solution (pH 6) was adjusted to the pH shown in parentheses with sodium hydroxide.

The same test was conducted with 90% propylene glycol, 90% N-methyl-2-pyrrolidone (NMP), and 90% dimethylsulfoxide (DMSO) aqueous solution. However, compared with the absorbance immediately after coating, the UV absorption effect persistence of 50% or more was not shown.

(Example 5)
The quartz plate was attached to UV-STAR MICROPLATE, 96 Well. Well number B9-B11, C9-C11, D9-D11 including 9 points, 4 cm x 4 cm (16 cm ² ), 0.5% cinolin and 0.1% sodium hyaluronate 50% 1,3-butanediol. 27 mg of an aqueous solution was thinly applied. The plate was left at room temperature for 24 hours. The average value of the absorbance at 334 nm was measured at 9 points of B9-B11, C9-C11, and D9-D11 immediately after coating and after 24 hours. As a result, the UV absorption effect persistence was maintained at 80% or more after 24 hours compared with the absorbance immediately after coating.

(Reference example 1-1)
Cinoline 9 mg was dissolved in 0.1 M phosphate buffer (sodium dihydrogen phosphate+disodium hydrogen phosphate; pH 7) 9 ml, and heated at 50° C. for 2 weeks under light shielding. The solution before the reaction was colorless and transparent. On the other hand, the solution after the reaction for 2 weeks was colored brown. The absorbance at 400 nm increased before and after the reaction.

(Reference example 1-2)
Cinoline (9 mg) was dissolved in water (9 ml) (pH 3), and the mixture was heated at 50°C for 2 weeks in the dark. The solution before the reaction was colorless and transparent. On the other hand, the solution after the reaction for 2 weeks was colored brown. The absorbance at 400 nm increased before and after the reaction.

(Reference example 2)
<HPLC analysis conditions>
Column: YMC-Pack ODS-AQ S-5 μM, 12 nm 250×4.6 mm I.D. D.
Flow rate: 0.5 ml/min Temperature: 30°C
Detection: PDA (334nm)
Eluent: 0.1% formic acid solution in water Retention time of cinolin: around 8 minutes Dissolve 9 mg of shinorine in 9 ml of 0.1 M phosphate buffer (sodium dihydrogen phosphate + disodium hydrogen phosphate; pH 7), and protect from light 50 Heated at 0°C for 2 weeks. The cynoline concentration before and after the reaction was measured by HPLC. As a result, the content of cynoline decreased.

(Reference example 3)
A quartz plate was attached to UV-STAR MICROPLATE, 96Well, and 3% x 3 cm (9 cm ² ) containing 9 points of well numbers B9-B11, C9-C11 and D9-D11 contained 90% of 0.5% shinoline. 19 mg of 3-butanediol aqueous solution was applied thinly. The plate was left at room temperature for 24 hours. The average value of the absorbance at 334 nm was measured at 9 points of B9-B11, C9-C11 and D9-D11 immediately after coating and after 24 hours. As a result, the absorbance at 334 nm decreased to less than 50%.

(Reference example 4)
1.5 mg of Porphyra-334 was dissolved in 1.5 ml of water (pH 3), and the mixture was heated at 50° C. for 17 days in the dark. The solution before the reaction was colorless and transparent. On the other hand, the solution after reacting for 17 days was colored brown.

(Reference example 5)
1.5 mg of Porphyra-334 was dissolved in 1.5 ml of water (pH 3), and the mixture was heated at 50° C. for 17 days in the dark. The Porphyra-334 concentration before and after the reaction was measured under the HPLC conditions described in Reference Example 2. As a result, the content of Porphyra-334 was reduced. The retention time of Porphyra-334 was around 12 minutes.

(Reference example 6)
Porphyra-334 (1.5 mg) was dissolved in water (pH 3), and the mixture was heated at 50°C for 17 days in the dark. The solution before the reaction was colorless and transparent. On the other hand, the solution after the reaction for 2 weeks was colored brown.

According to the present invention, a solution containing stabilized MAA can be provided. Such a solution can be used for producing an ultraviolet absorbing composition. Therefore, the present invention can be used in the fields of protection of various industrial products and medical devices from deterioration due to ultraviolet rays, and cosmetics, quasi-drugs and pharmaceuticals.

Claims (8)

A method for suppressing the decomposition of shinorin in a solution,
(A) cynoline, and (b) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, Group consisting of methionine, serine, glucose, trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, and ergothioneine. One or more substances selected from
Is mixed with a solvent. A method for suppressing the coloring of a shinorin solution,
(A) cynoline, and
(B) Glutathione, sodium disulfite, albumin, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, trehalose, sorbitol, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride One or more substances selected from the group consisting of salts, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate;
Is mixed with a solvent. A method for suppressing the decomposition of shinorin in a solution, and a method for suppressing coloration,
(A) cinolin, and (b) glutathione, sodium disulfite, albumin, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, methionine, trehalose, sorbitol, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, One or more substances selected from the group consisting of lysine hydrochloride, mannitol, sodium hyaluronate, and ergothioneine;
Is mixed with a solvent. A method for suppressing a decrease in the UV absorption capacity of shinorin when a shinorin solution is applied in a thin film state,
A solvent containing (a) cinolin and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A method characterized by mixing. A composition for suppressing the decomposition of shinorin in a solution,
(A) Cinoline,
(B) α-glucosyl hesperidin, erythorbic acid, α-glucosyl rutin, glutathione, sodium disulfite, albumin, gallic acid, EDTA, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, methionine, serine, glucose, One or more selected from the group consisting of trehalose, sorbitol, allose, β-cyclodextrin, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride, glucosamine hydrochloride, mannitol, sodium hyaluronate, and ergothioneine. And a solvent (c). A composition for suppressing coloration of a shinorin solution,
(A) Cinoline,
(B) Glutathione, sodium disulfite, albumin, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, arginine, cysteine, methionine, trehalose, sorbitol, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride A composition comprising one or more substances selected from the group consisting of salts, mannitol, sodium hyaluronate, ergothioneine, and tocopherol acetate, and (c) a solvent. A composition for suppressing decomposition and coloring of a shinorin solution,
(A) Cinoline (b) Glutathione, sodium disulfite, albumin, sodium metaphosphate, sodium citrate, aspartic acid, glutamic acid, methionine, trehalose, sorbitol, potassium sorbate, sodium benzoate, sodium parahydroxybenzoate, lysine hydrochloride A composition comprising one or more substances selected from the group consisting of salts, mannitol, sodium hyaluronate, and ergothioneine, and (c) a solvent. A composition for suppressing lowering of UV absorption capacity of shinorin when the shinorin solution is applied in a thin film state,
A solvent containing (a) cinolin and (c) one or more substances selected from the group consisting of glycerol, 1,4-butanediol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, and pentaethylene glycol. A composition comprising.