Composition containing hyaluronic acid or its salt and indigo carmine

Abstract

Provided is a composition having excellent stability and containing hyaluronic acid or a salt thereof and indigocarmine. The composition contains: hyaluronic acid or a salt thereof; indigocarmine; L-methionine; and citric acid or a salt thereof.

Description

[Technical Field]

[0001] The present invention relates to a composition comprising hyaluronic acid or a salt thereof and indigo carmine. [Background technology]

[0002] We live in an age where one in two Japanese people will develop cancer and one in three will die from it. Advances in testing and treatment methods mean that cancer is a curable disease if detected and treated early. From a medical economic perspective, early cancer treatment of the gastrointestinal tract, particularly endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD), is an extremely effective method, and demand for these procedures is expected to continue to increase. To perform EMR and ESD easily and safely, an appropriate amount of a highly viscous solution is injected into the submucosa at the site of the lesion to be resected or dissected, maintaining the mucosal protuberance formed, improving the ease of resection or dissection.

[0003] For large lesions or when fractional EMR is expected to require a long treatment time, sodium hyaluronate is used as a highly viscous substance suitable for forming and maintaining mucosal elevations (see, for example, the Gastrointestinal Endoscopy Handbook, Revised 2nd Edition, pp. 400-412). Products using sodium hyaluronate are commercially available, such as "Mucoup®" and "Keismart®," which are clear, colorless, and slightly viscous liquids filled in glass vials (see the package insert for "Mucoup," a highly controlled medical device for endoscopic submucosal injection). International Publication Nos. 2012 / 118190 and 2012 / 118192 also disclose technologies related to highly stable aqueous solutions containing hyaluronic acid or its salts.

[0004] As a preliminary step for EMR and ESD, for example, to facilitate visualization of the submucosal layer, it is necessary to add the coloring agent indigo carmine to a sodium hyaluronate solution at a concentration sufficient to make blood vessels and muscle layers transparent (see Handbook of Gastrointestinal Endoscopy, Revised 2nd Edition, pp. 400-412). Indigo carmine is known to be easily oxidized and to fade due to the effects of oxygen in the air and light. International Publication No. 2012 / 018723 discloses a technology related to a liquid preparation containing indigo carmine in a stable state. Summary of the Invention

[0005] When adding indigo carmine to sodium hyaluronate, the indigo carmine solution is dispensed from the glass vial into a disposable syringe and diluted with saline as needed.

[0006] The package insert for "Mucoup," a submucosal injection material for endoscopy that is a highly regulated medical device, states that when mixed with indigo carmine solution, "stability beyond five hours after mixing has not been confirmed," and the mixed solution can only be used for up to five hours.

[0007] In medical settings, there is a need to reduce the burden of drug preparation associated with these tasks, the risk of needlestick injuries and drug mix-ups, and the risk of indigo carmine decomposition. However, no method is known for stabilizing both hyaluronic acid or its salt and indigo carmine in an aqueous solution containing hyaluronic acid or its salt and indigo carmine.

[0008] Therefore, an object of the present invention is to provide a composition containing hyaluronic acid or a salt thereof and indigo carmine, which has excellent stability.

[0009] The present inventors have conducted extensive research to solve the above problems, and as a result, have found that the above problems can be solved by a composition containing hyaluronic acid or a salt thereof, indigo carmine, L-methionine, and citric acid or a salt thereof, thereby completing the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0010] An embodiment of the present invention will be described below, but the present invention is not limited to the following embodiment.

[0011] In this specification, the range "X to Y" means "X or more and Y or less." Unless otherwise specified, operations and measurements of physical properties are carried out at room temperature (1 to 30°C).

[0012] <Composition> One aspect of the present invention is a composition comprising hyaluronic acid or its salt, indigo carmine, L-methionine, and citric acid or its salt.According to this aspect, it is possible to provide a composition comprising hyaluronic acid or its salt and indigo carmine, which has excellent stability.

[0013] As mentioned above, there are many techniques for enhancing the stability of hyaluronic acid or its salt (for example, International Publication No. 2012 / 118190 and International Publication No. 2012 / 118192), or for enhancing the stability of indigo carmine (for example, International Publication No. 2012 / 018723).However, when hyaluronic acid or its salt and indigo carmine are mixed, there is no known method for stabilizing both hyaluronic acid or its salt and indigo carmine.

[0014] The present inventors have surprisingly found that the stability of hyaluronic acid or a salt thereof and indigo carmine is improved by combining L-methionine with citric acid or a salt thereof.

[0015] (Hyaluronic acid or its salts) The composition of this form contains hyaluronic acid or a salt thereof.

[0016] Hyaluronic acid is composed of the basic structure (repeating unit) of GlcUA-GlcNAc, which is the combination of glucuronic acid (GlcUA) and N-acetylglucosamine (GlcNAc).There is no particular limitation on the salt of hyaluronic acid, as long as it is in the form of a pharmaceutically or pharmacologically acceptable salt.The salt of hyaluronic acid can be exemplified by sodium hyaluronate, potassium hyaluronate, zinc hyaluronate, calcium hyaluronate, magnesium hyaluronate, ammonium hyaluronate, etc.

[0017] The hyaluronic acid or its salt according to this embodiment can be any hyaluronic acid or its salt that can be used in endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD).

[0018] In a preferred embodiment, the hyaluronic acid or salt thereof is sodium hyaluronate.

[0019] The concentration of hyaluronic acid or its salt in the composition can be adjusted appropriately depending on the application.When the composition is used for EMR and ESD, the concentration of hyaluronic acid or its salt is preferably 0.1 to 1.0 w / v%, more preferably 0.2 to 0.4 w / v%, from the viewpoint of ease of pushing during injection and formation and maintenance of mucosal protuberance.

[0020] The weight-average molecular weight of hyaluronic acid or its salt is, for example, 500,000 or more, and from the viewpoint of viscosity, it is preferably 600,000 or more, more preferably 600,000-1,200,000.If it is in this range, it is suitable for use in EMR and ESD.The weight-average molecular weight of hyaluronic acid or its salt can be calculated by measuring the limiting viscosity according to the 17th revision of the Japanese Pharmacopoeia, General Test Method, Section 2.53, Viscosity Measurement Method, and using Laurent's formula.

[0021] Hyaluronic acid and its salts may be naturally occurring or synthetic, and commercially available products may also be used.

[0022] (indigo carmine) The composition of this form contains indigo carmine (chemical name: disodium 3,3'-dioxo-2,2'-bis-indolidene-5,5'-disulfonate). Indigo carmine may be in the cis or trans form, preferably the trans form.

[0023] The concentration of indigo carmine in the composition is preferably 0.0002 to 0.012 w / v %, and more preferably 0.001 to 0.004 w / v %, from the viewpoint of visibility during EMR or ESD.

[0024] The indigo carmine according to the present invention may be synthesized by referring to a conventionally known method, or a commercially available product may be used.

[0025] (L-methionine and citric acid or its salts) The composition of this form contains L-methionine and citric acid or a salt thereof. The combination of L-methionine and citric acid or a salt thereof can stabilize hyaluronic acid or a salt thereof and indigo carmine.

[0026] In this specification, citric acid and its salts include hydrates thereof.

[0027] Examples of citric acid or a salt thereof include citric acid, sodium citrate, disodium citrate, trisodium citrate, and hydrates thereof. From the viewpoint of being able to more effectively exhibit the effects of the present invention, the citric acid or a salt thereof is preferably sodium citrate or sodium citrate hydrate.

[0028] The concentrations of L-methionine and citric acid or a salt thereof in the composition can be adjusted appropriately depending on the amount of hyaluronic acid or a salt thereof used.

[0029] The content of L-methionine in the composition is, for example, 0.25 to 50 parts by mass relative to 100 parts by mass of hyaluronic acid or a salt thereof, and from the viewpoint of being able to more effectively exhibit the effects of the present invention, it is preferably 0.5 to 50 parts by mass, more preferably 1 to 30 parts by mass, and even more preferably 1 to 15 parts by mass.Furthermore, the content of L-methionine in the composition is, for example, 0.25 to 12.5 parts by mass, more preferably 1.25 to 5 parts by mass relative to 100 parts by mass of hyaluronic acid or a salt thereof, from the viewpoint of being able to more effectively exhibit the effects of the present invention.

[0030] The content of citric acid or its salt in the composition is, for example, 0.25 to 50 parts by mass relative to 100 parts by mass of hyaluronic acid or its salt, and from the viewpoint of being able to more effectively exhibit the effects of the present invention, it is preferably 0.5 to 50 parts by mass, more preferably 1 to 30 parts by mass, and even more preferably 10 to 20 parts by mass. Also, the content of citric acid or its salt in the composition is, for example, 0.25 to 25 parts by mass, more preferably 1.25 to 25 parts by mass relative to 100 parts by mass of hyaluronic acid or its salt, from the viewpoint of being able to more effectively exhibit the effects of the present invention.

[0031] Commercially available L-methionine and citric acid or a salt thereof can be used.

[0032] (Other ingredients) The composition of this form may contain other components as needed, such as a solvent, a pH adjusting agent, a buffer, and a tonicity adjusting agent.

[0033] The solvent may be water. Examples of water include distilled water, tap water, purified water, sterile purified water, water for injection, distilled water for injection, etc. The definition of water is based on the 17th edition of the Japanese Pharmacopoeia.

[0034] Examples of pH adjusters include bases such as sodium hydroxide and potassium hydroxide; inorganic acids such as hydrochloric acid; and organic acids such as lactic acid, acetic acid, succinic acid, and malic acid.

[0035] Examples of the buffering agent include phosphoric acid, a salt thereof, or a hydrate thereof (such as sodium hydrogen phosphate hydrate, trisodium phosphate hydrate, and sodium dihydrogen phosphate hydrate), acetic acid, a salt thereof, or a hydrate thereof.

[0036] The concentrations of the pH adjuster and buffer are not particularly limited and can be adjusted appropriately to achieve the desired pH.

[0037] Examples of the isotonicity agent include non-ionic isotonicity agents such as mannitol, sorbitol, inositol, glucose, propylene glycol, glycerol, etc., and ionic isotonicity agents such as sodium chloride, etc. The isotonicity agent is preferably sodium chloride.

[0038] The concentration of the isotonic agent can be adjusted as appropriate. The osmotic pressure ratio of the composition is, for example, 0.9 to 1.2 (ratio to physiological saline).

[0039] (pH of composition) The pH of the composition of this embodiment is, for example, 6 to 8, and preferably 7 to 8. The pH of the composition can be adjusted using the above-mentioned pH regulator and / or buffer, if necessary. The pH of the composition can be confirmed using a pH meter.

[0040] (Method of preparing the composition) The method for preparing the composition of this form is not particularly limited, and the composition can be prepared by mixing hyaluronic acid or a salt thereof, indigo carmine, L-methionine, and citric acid or a salt thereof, and other ingredients as necessary.

[0041] Specifically, ingredients other than hyaluronic acid or its salt and indigo carmine are dissolved in water (e.g., water for injection). If necessary, the pH of the solution is adjusted to 7 to 8 using a pH adjuster and / or buffer. After dissolving hyaluronic acid or its salt in this solution, indigo carmine is dissolved, and water is added as needed to prepare the composition. When preparing the composition, the solution may be heated appropriately to improve solubility depending on the concentration of the ingredients used.

[0042] The composition can be prepared in the atmosphere (air) or in an inert gas atmosphere, such as nitrogen gas.

[0043] The prepared composition is appropriately sterilized. Examples of sterilization methods include filtration sterilization, high-pressure steam sterilization, etc. The composition may be sterilized after being filled into a medical container.

[0044] (Use of the composition) The composition of this form can be used for forming and maintaining a mucosal prominence at a tumor site under endoscopic surgery. Thus, in one embodiment, the composition of this form is for submucosal local injection, preferably for endoscopic mucosal resection and / or endoscopic submucosal dissection. The composition of this form may be diluted appropriately to achieve a desired viscosity depending on the situation.

[0045] <Package> One embodiment of the present invention is a package comprising a medical container filled with the composition and enclosed in an oxygen-barrier container together with an oxygen scavenger. By enclosing the medical container filled with the composition in an oxygen-barrier container together with an oxygen scavenger, the effects of oxygen can be suppressed, and storage stability can be further improved. In particular, deterioration (decomposition and discoloration) of indigo carmine can be further suppressed.

[0046] The medical container filled with the composition and the oxygen scavenger can be enclosed in an oxygen-barrier container according to a conventional method. The encapsulation can be carried out in the atmosphere (air), in an inert gas atmosphere, or in a mixed gas atmosphere of air and an inert gas. Furthermore, the encapsulation can be carried out after replacing the atmosphere inside the oxygen-barrier container with an inert gas.

[0047] (Medical containers) The shape, size, etc. of the medical container are not particularly limited and can be appropriately selected depending on the amount of the composition to be filled, the application, the form of use, etc.

[0048] The material for forming the medical container is, for example, a resin, preferably a resin with excellent steam sterilization resistance, transparency, moldability, and versatility. Examples of such resins include polyesters such as polypropylene, polyethylene, polystyrene, polyamide, polycarbonate, polyvinyl chloride, poly-(4-methylpentene-1), acrylic resin, acrylonitrile-butadiene-styrene copolymer, and polyethylene terephthalate, and cyclic polyolefins such as cyclic olefin polymer (COP) and cyclic olefin copolymer (COC).

[0049] Examples of cyclic olefins constituting cyclic polyolefins include monocyclic cyclic olefins, polycyclic cyclic olefins, polycyclic cyclic olefins having a bridged structure, etc. One or more types of cyclic olefins can be used.

[0050] Examples of cyclic olefins include substituted or unsubstituted norbornene-based monomers, substituted or unsubstituted tetracyclododecene-based monomers, substituted or unsubstituted dicyclopentadiene-based monomers, and cyclic polyolefins having a cyclohexyl group or a phenyl group in the side chain.

[0051] When the cyclic olefin copolymer is a copolymer of a cyclic olefin and an acyclic olefin, examples of the acyclic olefin include ethylene, propylene, butene, isobutylene, methylpentene, etc. One or more types of the acyclic olefin can be used.

[0052] The cyclic polyolefin may be synthesized or a commercially available product may be used, such as "ZEONEX (registered trademark)" (manufactured by Zeon Corporation), "ZEONOR (registered trademark)" (manufactured by Zeon Corporation), "APEL (registered trademark)" (manufactured by Mitsui Chemicals, Inc.), or "TOPAS (registered trademark)" (manufactured by Polyplastics Co., Ltd.).

[0053] Examples of the form of the medical container include a syringe, a vial, a bag, an ampoule, a cartridge, a bottle, etc., and preferably a syringe or a vial, more preferably a syringe. Therefore, the medical container according to this embodiment is preferably a plastic pre-filled syringe.

[0054] The syringe may have a gasket made of an elastomer. Examples of elastomers include, but are not limited to, natural rubber, isoprene rubber, chlorinated or brominated butyl rubber, chloroprene rubber, nitrile-butadiene rubber, styrene-butadiene rubber, and silicone rubber (especially vulcanized rubbers); thermoplastic elastomers such as styrene elastomers, hydrogenated styrene elastomers, polyvinyl chloride elastomers, olefin elastomers, polyester elastomers, polyamide elastomers, and polyurethane elastomers; and mixtures of styrene elastomers with polyolefins such as polyethylene, polypropylene, polybutene, and α-olefin polymers, oils such as liquid paraffin and process oil, and powdered inorganic materials such as talc, cast, and mica. The elastomer may be a single type or a mixture of two or more types.

[0055] The elastomer is preferably butyl rubber or a thermoplastic elastomer, more preferably butyl rubber or a styrene-based elastomer, from the viewpoints that it has elastic properties and can be sterilized by gamma rays, electron beams, and high-pressure steam.

[0056] The gasket that constitutes the syringe is preferably in a form that fits tightly against the inside of the barrel, seals the rear end opening side of the internal space of the barrel, is easily slidable within the barrel, and allows a plunger to be attached to the rear end.

[0057] The composition can be filled into a medical container according to a conventional method. The filling can be carried out in the atmosphere (air), in an inert gas atmosphere, or in a mixed gas atmosphere of air and an inert gas. Examples of inert gases that can be used include nitrogen gas, argon gas, and carbon dioxide gas.

[0058] The medical container filled with the composition is preferably sterilized. The sterilization method is not particularly limited, and conventionally known methods such as high-pressure steam sterilization can be used. When high-pressure steam sterilization is used as the sterilization method, steam can be used as the sterilization medium for high-pressure steam sterilization. A mixed gas of inert gas and air or air can be used as the pressurized medium for high-pressure steam sterilization. That is, high-pressure steam sterilization may be performed using steam as the sterilization medium and air as the pressurized medium, or using steam as the sterilization medium and an inert gas as the pressurized medium, or using steam as the sterilization medium and a mixed gas of air and an inert gas as the pressurized medium.

[0059] Examples of inert gases used in high-pressure steam sterilization include nitrogen gas, argon gas, and carbon dioxide gas, with nitrogen gas being preferred because it is available at low cost.

[0060] The sterilization conditions, such as temperature, pressure, and time, used in high-pressure steam sterilization may be the same as those normally used in sterilizing medical containers. For example, the temperature is usually 100 to 129°C, and preferably 115 to 124°C.

[0061] The method for manufacturing a medical container is not particularly limited, and known methods such as injection molding, extrusion molding, blow molding, rotational molding, blow molding, transfer molding, press molding, and solution casting can be appropriately selected depending on the type and shape of the material forming the medical container.

[0062] (oxygen absorber) There are no particular limitations on the oxygen scavenger, as long as it does not pose problems such as toxicity in use and can efficiently absorb oxygen to reduce the amount of oxygen in the atmosphere.

[0063] Examples of oxygen scavengers include iron compounds such as iron hydroxide, iron oxide, and iron carbide; oxygen scavengers based on inorganic substances such as nitrites and metal halides; oxygen scavengers based on organic substances such as ascorbic acid and polyphenols; and oxygen scavengers that utilize the enzymatic action of glucose and glucose oxidase.

[0064] The oxygen absorber may be one that absorbs oxygen and carbon dioxide gas at the same time, or one that absorbs oxygen and releases carbon dioxide gas.

[0065] The form of the oxygen absorber is not particularly limited as long as it is non-toxic and exhibits excellent oxygen absorbing properties. For example, an oxygen-permeable container in which the above-mentioned substance having oxygen absorbing properties is enclosed, a film kneaded with an oxygen absorber, a molded container using an oxygen absorber, a laminated structure in which both surface layers are oxygen-impermeable layers and an oxygen-absorbing layer is disposed inside the layers, and oxygen is absorbed from the side of the laminate, etc. can be used.

[0066] Commercially available oxygen absorbers include Ageless (registered trademark) (manufactured by Mitsubishi Gas Chemical Company, Inc.), Moduran (manufactured by Nippon Kayaku Food Techno Co., Ltd.), and Secur (registered trademark) (manufactured by Nisso Fine Co., Ltd.).

[0067] (Oxygen barrier container) The oxygen barrier container is not particularly limited as long as it is a container that can enclose the medical container and is non-toxic. The oxygen barrier container may be a soft container or a hard container. In addition, the oxygen barrier container may have a light-shielding ability.

[0068] The oxygen permeability of the material forming the oxygen barrier container is preferably 1.0 cm when measured at 23°C and 50% RH, from the viewpoint of further demonstrating the effects of the present invention. 3 / m 2 ·day·atm or less, and more preferably 0.3cm 3 / m 2 ·day·atm or less, and more preferably 0.1cm 3 / m 2 The oxygen permeability can be measured in accordance with JIS K 7126-2:2006 under conditions of 23°C and 50% RH using an oxygen permeability measuring device (OX-TRAN (registered trademark) MODEL2 / 22 manufactured by MOCON).

[0069] The material for forming the oxygen barrier container is not particularly limited as long as it has a certain degree of strength and hardness and oxygen barrier properties. Examples of such materials include metal foils such as aluminum, gold, and silver, metal-deposited films and sheets such as aluminum, gold, and silver, inorganic-deposited films and sheets such as SiOx, films and sheets formed from polyvinylidene chloride, polyvinylidene chloride-polyvinyl chloride, polyvinylidene chloride-acrylic acid ester copolymers, ethylene-vinyl alcohol copolymers, high-density polyethylene, and the like.

[0070] Examples of materials for the substrate of metallized films, metallized sheets, inorganic vapor-deposited films and inorganic vapor-deposited sheets include polyolefin resins such as polypropylene and polyethylene, polyamide, polyvinyl chloride, polyester, and polystyrene / polypropylene resin.

[0071] The material forming the oxygen barrier container may be a laminate. At least one layer of the laminate contains the above-mentioned material having oxygen barrier properties. The materials of the other layers constituting the laminate are not particularly limited.

[0072] When oxygen barrier containers are formed from films or sheets, the packaging container is often formed by folding the film or sheet and heat-sealing the periphery, so it is preferable that the part that will become the innermost surface when folded is formed from at least a heat-sealable adhesive resin layer (for example, polyolefin resins such as polypropylene and polyethylene, polystyrene / polypropylene resin, ethylene-vinyl acetate resin, ethylene-acrylic acid resin, polyvinyl chloride resin, or various thermoplastic elastomers).

[0073] Specific examples of materials for oxygen barrier containers include, but are not limited to: · Multilayer film or sheet made of polypropylene (PP) / silica-deposited polyethylene terephthalate (PET) / polypropylene (PP) laminated in this order; · Multilayer film or sheet made by laminating biaxially oriented polyamide (OPA) / polyethylene (PE) / aluminized PET / polyethylene (PE) in this order; Multilayer film or sheet made of OPA / PE / aluminized PET / PE laminated in this order; Multilayer film or sheet made by laminating OPA / PE / aluminum foil / PE / PE in this order; Multilayer film or sheet made by laminating OPA / PE / aluminum foil / PE / PET / PE in this order; Multilayer film or sheet made of PET / PE / aluminized PET / PE / ethylene-vinyl acetate copolymer (EVA) / PE laminated in this order; Multilayer film or sheet made of polyvinylidene chloride / PE / aluminized PET / PE laminated in this order; · Multilayer film or sheet made by laminating PET / aluminized ethylene-vinyl alcohol copolymer (EVOH) / PE in this order; · Multilayer film or sheet made of biaxially oriented nylon (ONY) / EVOH / PE laminated in this order; Multilayer film or sheet made by laminating PET / aluminized EVOH / a blend of the above resins in that order Multilayer film or sheet made by laminating PP / EVOH / PP in this order Examples include:

[0074] Examples of the oxygen barrier container include blister containers, bags, tubes, etc. When the oxygen barrier container is a blister container, a combination of a top film (for example, a multilayer film of PET / aluminum-deposited EVOH / a blend of the above resin) and a bottom film (for example, a multilayer film of PP / EVOH / PP) can be used.

[0075] The method for producing the oxygen barrier container is not particularly limited, and any known method can be appropriately selected depending on the type of material from which the oxygen barrier container is made, the shape, etc.

[0076] <Stabilization method> The composition of the present invention contains L-methionine and citric acid or its salt, thereby improving the stability of hyaluronic acid or its salt and indigo carmine.Therefore, one aspect of the present invention is a method for stabilizing hyaluronic acid or its salt and indigo carmine, which comprises mixing hyaluronic acid or its salt, indigo carmine, L-methionine, and citric acid or its salt.

[0077] The specific explanation of the stabilization method has been given above, so a detailed explanation will be omitted here. [Example]

[0078] The effects of the present invention will be explained using the following examples and comparative examples. However, the technical scope of the present invention is not limited to the following examples. Unless otherwise specified, "%" and "parts" mean "% by mass" and "parts by mass", respectively.

[0079] <Example 1, Reference Example 1, and Comparative Examples 1 to 3> Under a nitrogen atmosphere, L-methionine, sodium citrate hydrate, sodium chloride, trisodium phosphate hydrate, and sodium hydrogen phosphate hydrate were dissolved in an appropriate amount of water for injection in the amounts listed in Table 1 below, and then the pH was adjusted to 7-8 using sodium dihydrogen phosphate hydrate. Sodium hyaluronate with a weight-average molecular weight of approximately 1,000,000 was added in small amounts to this solution and dissolved, followed by the addition of indigo carmine and water for injection to a total volume of 1,000 mL to prepare a composition. 20 mL of this composition was filled into a syringe, sealed, and then sterilized by high-pressure steam (121°C, 4 minutes) using nitrogen gas as the pressure medium. The sterilized syringe and an oxygen absorber (Ageless®, manufactured by Mitsubishi Gas Chemical Company, Inc.) were then sealed in an oxygen-barrier container (a bag made of aluminum-deposited film) to prepare a package. Since Reference Example 1 does not contain indigo carmine, the package was sealed in an oxygen-barrier container without the addition of an oxygen absorber.

[0080] [Table 1]

[0081] <Test Example 1> The packaged bodies of Example 1, Reference Example 1 and Comparative Examples 1-3 are stored at 60°C for 2 weeks (protected from light), and the weight-average molecular weight of sodium hyaluronate is measured at the beginning (after sterilization), after 1 week and after 2 weeks.The weight-average molecular weight is measured by measuring the limiting viscosity according to the viscometric method of Section 2.53 of the Japanese Pharmacopoeia General Test Method, 17th Edition, and calculated by Laurent's formula.The change rate (%) of the weight-average molecular weight at the beginning (after sterilization) is also shown (Table 2).

[0082] [Table 2]

[0083] <Test Example 2> The packages of Example 1 and Comparative Examples 1 to 3 were stored at 60°C for 2 weeks (protected from light), and the indigo carmine content was measured at a wavelength of 254 nm at the start (after sterilization), and after 1 week and 2 weeks, based on the literature of Bailey JE (J. Assoc. Off. Anal Chem, 63, 565 (1980)). The percentage change (%) when the content at the start (after sterilization) is set to 100% is also shown (Table 3).

[0084] [Table 3]

[0085] As shown in Tables 2 and 3, the composition of Example 1 containing both L-methionine and sodium citrate hydrate shows a smaller decrease in the weight-average molecular weight of sodium hyaluronate and the content of indigo carmine compared to the composition of Comparative Example.In particular, it can be seen that the stability of sodium hyaluronate is comparable to that of the composition of Reference Example 1 that does not contain indigo carmine.

[0086] On the other hand, the composition of Comparative Example 1, which does not contain L-methionine and sodium citrate hydrate, not only showed a decrease in the content of indigo carmine, but also a significant decrease in the weight-average molecular weight of sodium hyaluronate.The composition of Comparative Example 2, which does not contain sodium citrate hydrate, showed a decrease in the content of indigo carmine similar to that of the composition of Example 1, but a significant decrease in the weight-average molecular weight of sodium hyaluronate.The composition of Comparative Example 3, which does not contain L-methionine, showed a significant decrease in the weight-average molecular weight of sodium hyaluronate and the content of indigo carmine.

[0087] <Examples 2 to 7> Under a nitrogen atmosphere, L-methionine, sodium citrate hydrate, sodium chloride, and trisodium phosphate hydrate were dissolved in an appropriate amount of water for injection in the amounts listed in Table 4 below, and then the pH was adjusted to 7-8 using sodium dihydrogen phosphate hydrate. Sodium hyaluronate with a weight-average molecular weight of approximately 1,000,000 was added in small amounts to this solution and dissolved, followed by the addition of indigo carmine and water for injection to a total volume of 1,000 mL to prepare a composition. Five mL of this composition was filled into a syringe, sealed, and then the syringe was subjected to autoclaving (121°C, 1.5 minutes) using nitrogen gas as the pressure medium. The sterilization time for autoclaving was set to achieve the same F0 value as in Example 1, Reference Example 1, and Comparative Examples 1-3. The sterilized syringe and an oxygen absorber ("AGELESS (registered trademark)" manufactured by Mitsubishi Gas Chemical Company, Inc.) were then sealed in an oxygen-barrier container (a bag made of aluminum-deposited film) to prepare a package.

[0088] [Table 4]

[0089] <Test Example 3> The packaged body of Examples 2 to 7 is stored at 60 ℃ for 2 weeks (protected from light), and the weight-average molecular weight of sodium hyaluronate is measured at the beginning (after sterilization), after 1 week, and after 2 weeks.The weight-average molecular weight is measured by measuring the limiting viscosity according to the viscometric method of the Japanese Pharmacopoeia, General Test Method, Section 2.53, 17th Edition, and calculated by Laurent's formula.In addition, the change rate (%) is also shown (Table 5), assuming that the average molecular weight at the beginning (after sterilization) is 100%.

[0090] [Table 5]

[0091] <Test Example 4> The packages of Examples 2 to 7 were stored at 60°C for 2 weeks (protected from light), and the indigo carmine content was measured at a wavelength of 254 nm at the start (after sterilization), one week later, and two weeks later using liquid chromatography based on Bailey JE's literature (J. Assoc. Off. Anal Chem, 63, 565 (1980)). The percentage change (%) when the content at the start (after sterilization) is taken as 100% is also shown (Table 6).

[0092] [Table 6]

[0093] As shown in Tables 5 and 6, Example 2, which has the same formulation as Example 1, showed the same changes over time in the weight-average molecular weight of sodium hyaluronate and the content of indigo carmine, even when the syringe size was changed.

[0094] Furthermore, compared to Example 2, Example 3, in which the contents of both L-methionine and sodium citrate hydrate were reduced, and Example 4, in which only the content of sodium citrate hydrate was reduced, both showed that the decrease in the weight-average molecular weight of sodium hyaluronate was suppressed, as in Example 2.

[0095] Furthermore, compared to Example 2, Example 5, in which the content of sodium citrate hydrate was increased, and Examples 6 and 7, in which the content of L-methionine was increased and the content of sodium citrate hydrate was reduced, were found to suppress the decrease in the weight-average molecular weight of sodium hyaluronate, as in Example 2.

[0096] This application is based on Japanese Patent Application No. 2020-162006 filed on September 28, 2020, the disclosure of which is incorporated herein by reference in its entirety.

Claims

1. A composition comprising hyaluronic acid or a salt thereof, indigo carmine, L-methionine, and citric acid or a salt thereof, The weight average molecular weight of the hyaluronic acid or its salt is 600,000 or more, The concentration of the hyaluronic acid or its salt is 0.1 to 0.5 w / v %, the concentration of the indigo carmine is 0.0002 to 0.012 w / v%, the content of the L-methionine is 0.25 to 50 parts by mass per 100 parts by mass of the hyaluronic acid or its salt, The composition, wherein the content of the citric acid or a salt thereof is 0.25 to 50 parts by mass per 100 parts by mass of the hyaluronic acid or a salt thereof.

2. The composition according to claim 1, wherein the concentration of the hyaluronic acid or its salt is 0.2 to 0.4 w / v%.

3. The composition according to claim 1 or 2, wherein the concentration of the indigo carmine is 0.001 to 0.004 w / v%.

4. The composition according to any one of claims 1 to 3, wherein the content of the L-methionine is 0.25 to 12.5 parts by mass per 100 parts by mass of the hyaluronic acid or its salt.

5. The composition according to any one of claims 1 to 4, wherein the content of the citric acid or its salt is 0.25 to 25 parts by mass per 100 parts by mass of the hyaluronic acid or its salt.

6. The composition according to any one of claims 1 to 5, wherein the weight-average molecular weight of the hyaluronic acid or its salt is 600,000 to 1,200,000.

7. The composition according to any one of claims 1 to 6, which is for submucosal local injection.

8. A package comprising a medical container filled with the composition according to any one of claims 1 to 7, and enclosed together with an oxygen scavenger in an oxygen barrier container.

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