JPWO2019146777A1 - Pharmaceutical preparation and method for producing the same - Google Patents

Abstract

Provided is a novel pharmaceutical preparation which has excellent storage stability and can be suitably used for, for example, desensitization therapy. A pharmaceutical preparation comprising a lyophilized composition containing at least an allergen protein and hyaluronic acid.

Description

  The present invention relates to a pharmaceutical preparation and a method for producing the same.

  Conventionally, the number of patients having allergic diseases caused by allergens such as pollen and mites has increased. As a treatment for such an allergic disease, a coping therapy using an antihistamine is mainly used.

  On the other hand, in recent years, desensitization therapy using an allergen protein has attracted attention as a radical treatment method for allergic diseases.

  As a preparation used for desensitization therapy, for example, an injection for subcutaneous injection is known. However, problems with hyposensitization therapy by subcutaneous injection include the risk of anaferaxy shock, the necessity of administration by healthcare professionals, the need for frequent long-term hospital visits, the pain of injections, and refrigerated storage. There is.

  To solve such a problem, a method using a freeze-dried agent to which a stabilizer or an excipient is added has been proposed. For example, Patent Document 1 proposes a pharmaceutical composition in which timothy turf pollen allergen is stabilized by freeze-drying a solution containing gelatin and mannitol or starch and mannitol as a stabilizer. In addition, for example, Patent Document 2 proposes a pharmaceutical composition in which a recombinant protein of cedar pollen major allergen is stabilized by freeze-drying a solution containing mannitol as a stabilizer and acetic acid as a pH regulator. I have. In addition, for example, Patent Document 3 proposes a pharmaceutical composition in which a recombinant protein of a major mite allergen is stabilized by freeze-drying a solution containing Macrogol 4000, polysorbate 80, and sucrose.

  However, the freeze-dried preparations of the prior art have a porous structure, so that they are hard and brittle, causing problems such as chipping of the preparation during transportation. Due to the change in properties, there has been a problem that the handleability when administering a short time after opening the sealed individual package is poor. Furthermore, the freeze-dried preparations of the prior art dissolve rapidly in the oral cavity, making it difficult to control the sensitization time of the allergen, and require further improvement for use in the required desensitization therapy. .

JP 2006-513269 A Patent No. 3932272 JP 2007-277094 A

  Under such circumstances, the present invention is excellent in storage stability, excellent in handling at the time of administration, and furthermore, has an optimal dissolution profile for sensitizing allergen protein to oral mucosa or skin. A main object of the present invention is to provide a novel pharmaceutical preparation which can be controlled and can be suitably used for, for example, desensitization therapy.

  The present inventor has conducted intensive studies in order to solve the above problems. As a result, a pharmaceutical preparation comprising a freeze-dried body of a composition containing at least an allergen protein and hyaluronic acid is excellent in storage stability, excellent in handling at the time of administration, and furthermore, the allergen protein is used for oral mucosa. Alternatively, it has been found that the dissolution profile can be controlled to an optimal dissolution profile for sensitizing the skin, and can be suitably used for, for example, desensitization therapy. The present invention has been completed by further study based on such knowledge.

That is, the present invention provides the following aspects of the invention.
Item 1. A pharmaceutical preparation comprising a lyophilized composition containing at least an allergen protein and hyaluronic acid.
Item 2. Item 2. The pharmaceutical preparation according to item 1, wherein the molecular weight of the hyaluronic acid is 1 × 10 ⁵ dalton or more.
Item 3. Item 3. The pharmaceutical preparation according to item 1 or 2, wherein the molecular weight of the hyaluronic acid is 3 × 10 ⁵ dalton or more.
Item 4. Item 4. The pharmaceutical preparation according to any one of Items 1 to 3, wherein the intrinsic viscosity of the hyaluronic acid measured by the following measurement method is 3 dL / g or more.
(Method of measuring intrinsic viscosity)
Hyaluronic acid (50 mg) was weighed and dissolved in a 0.2 mol / L sodium chloride solution to make 100 mL, and 10 mL, 15 mL, and 20 mL of this solution were weighed, and a 0.2 mol / L sodium chloride solution was added to each to 25 mL. These liquids are used as sample solutions. Viscosity measurement of each sample solution and 0.2 mol / L sodium chloride solution in accordance with the provisions of "2.53 Viscosity measurement method 1. First method Capillary viscometer method" of Japanese Pharmacopoeia 17 The specific viscosity is measured in an environment of 30.0 ± 0.1 ° C. by the method, and the reduced viscosity is calculated. A graph is drawn with the reduced viscosity on the vertical axis and the concentration of hyaluronic acid (g / 100 mL) on the horizontal axis, and the intrinsic viscosity is determined from the intersection of the straight line connecting each point and the vertical axis.
However, the specific viscosity and the reduced viscosity are obtained from the following equations.
Specific viscosity = (number of seconds of flowing down of sample solution / 0.2 mol / L second rate of flowing down of sodium chloride solution) -1
Reduced viscosity = specific viscosity / concentration of hyaluronic acid (g / 100 mL)
Item 5. Item 5. The pharmaceutical preparation according to Item 4, wherein the intrinsic viscosity of the hyaluronic acid measured by the above-mentioned measuring method is 8 dL / g or more.
Item 6. Item 6. The pharmaceutical preparation according to any one of Items 1 to 5, wherein the allergen protein contains at least one of a pollen allergen protein and a mite allergen protein.
Item 7. Item 7. The pharmaceutical preparation according to any one of Items 1 to 6, comprising a base material and the freeze-dried product laminated on the base material.
Item 8. Item 8. The pharmaceutical preparation according to any one of Items 1 to 7, wherein the freeze-dried body contains a polymer different from hyaluronic acid.
Item 9. Item 10. The pharmaceutical preparation according to any one of Items 1 to 8, wherein the freeze-dried product further contains at least one of sugar, polyhydric alcohol, and sugar fatty acid.
Item 10. Item 10. The pharmaceutical preparation according to any one of Items 1 to 9, which is used for sublingual administration.
Item 11. Item 10. The pharmaceutical preparation according to any one of Items 1 to 9, which is used for transdermal administration.
Item 12. Item 12. The method for producing a pharmaceutical preparation according to any one of Items 1 to 11,
Preparing a composition comprising at least an allergen protein, hyaluronic acid, and water,
Freeze-drying the composition;
A method for producing a pharmaceutical preparation, comprising:

  According to the present invention, it is excellent in storage stability, excellent in handling at the time of administration, and can be controlled to an optimal dissolution profile for sensitizing allergen proteins to oral mucosa or skin, for example, It is possible to provide a novel pharmaceutical preparation which can be suitably used for desensitization therapy. Further, according to the present invention, it is possible to provide a suitable method for producing the pharmaceutical preparation.

It is an example of the schematic diagram of the pharmaceutical preparation of this invention. It is an example of the schematic diagram of the pharmaceutical preparation of this invention.

  The pharmaceutical preparation of the present invention is characterized by comprising a lyophilized composition containing at least an allergen protein and hyaluronic acid. The pharmaceutical preparation of the present invention having such a constitution is excellent in storage stability, excellent in handling at the time of administration, and further sensitizes allergen protein to oral mucosa or skin. The dissolution profile can be controlled to an optimal dissolution profile, and can be suitably used for, for example, desensitization therapy.

  More specifically, the pharmaceutical preparation of the present invention has excellent allergen activity stability when stored in a room temperature environment, and for example, does not require refrigerated storage of the pharmaceutical preparation. In addition, while being a freeze-dried product, it is not brittle, has low hygroscopicity, and can be easily administered to the oral cavity such as under the tongue and to the site of application of the skin while maintaining a constant shape. In sensitization therapy or transdermal hyposensitization therapy, the dose of the allergen protein can be easily adjusted. In addition, accidental ingestion of the pharmaceutical preparation can be prevented. Furthermore, in the pharmaceutical preparation of the present invention, the molecular weight and content of hyaluronic acid, the type of other components (for example, other polymers, sugars, polyhydric alcohols, sugar fatty acids, and the like described below) incorporated as necessary, Arbitrarily controlling the dissolution time of the pharmaceutical preparation in the oral cavity and skin by adjusting the content (that is, controlling the optimal dissolution profile to sensitize the allergen protein to the oral mucosa and skin) In addition, it can exhibit sticking properties to the oral mucosa and reduce discomfort in the oral cavity due to residues of the pharmaceutical preparation. Hereinafter, the pharmaceutical preparation of the present invention and the production method thereof will be described in detail.

  The oral mucosa to which the pharmaceutical preparation of the present invention can be applied refers to mucous membranes other than teeth in the oral cavity such as the inner cheek, upper jaw, tongue, sublingual, and gums. In desensitization therapy, a pharmaceutical preparation is generally applied sublingually, and the pharmaceutical preparation of the present invention can also be suitably used for sublingual administration of an allergen protein.

  The skin to which the pharmaceutical preparation of the present invention can be applied is not particularly limited, and includes, for example, arms, chest, abdomen, back, buttocks, thighs, bulging skin, and the like. The soles are not preferred. Percutaneous desensitization therapy allows an allergen protein to directly act on dendritic cells present in the epidermis of the skin and is an efficient route of administration. It can be suitably used for dermal administration.

  The pharmaceutical preparation of the present invention includes a lyophilized composition containing at least an allergen protein and hyaluronic acid. The pharmaceutical preparation of the present invention contains the molecular weight and content of hyaluronic acid, and the type and content of other components (for example, other polymers, sugars, polyhydric alcohols, sugar fatty acids, and the like described below) to be added as necessary. By adjusting the pH, the adhesive property to the oral mucosa can be developed, and the freeze-dried body is used as an adhesive surface with the oral mucosa (for example, sublingually), and is adhered to the oral mucosa to obtain an administration site. Can be applied to any part of the oral mucosa without being limited to the sublingual area.In addition, it is possible to control the oral mucosa to an optimal dissolution profile for efficiently sensitizing the allergen protein to the oral mucosa. It can be suitably used for desensitization therapy.

  The freeze-dried body is obtained, for example, by freeze-drying a composition containing at least an allergen protein, hyaluronic acid, and water. The freeze-dried body has a form such as cotton or sponge, and usually has a water content of 5% by mass or less.

  In the present invention, “hyaluronic acid” is used in a concept including hyaluronic acid and a salt thereof. Therefore, “hyaluronic acid and its salt” may be simply referred to as “hyaluronic acid”. The salt of hyaluronic acid is not particularly restricted but includes, for example, sodium hyaluronate, potassium hyaluronate, magnesium hyaluronate, calcium hyaluronate and the like. In the present invention, hyaluronic acid and a salt thereof may be used alone or in a combination of two or more.

  When the pharmaceutical preparation of the present invention is to be applied to the oral cavity, the average molecular weight of hyaluronic acid, the intrinsic viscosity, the ratio of hyaluronic acid, and the like are described below so that stickiness to the oral mucosa is developed. adjust. In particular, when the pharmaceutical preparation of the present invention is used for oral desensitization therapy, the state in which the pharmaceutical preparation of the present invention is adhered to the oral mucosa is maintained for a predetermined time (for example, about 1 minute to about 20 minutes). It is desirable. Therefore, it is preferable that the pharmaceutical preparation of the present invention has an adhesive property to the oral mucosa (specifically, it does not rapidly dissolve even when applied to the oral mucosa).

The average molecular weight of hyaluronic acid is appropriately adjusted according to the sticking property and solubility of the desired pharmaceutical preparation to the oral mucosa, and for example, a range of about 1000 to 5 × 10 ⁶ daltons. For example, in the case of a pharmaceutical preparation having no sticking property to the oral mucosa and rapidly dissolving in the oral cavity (for example, dissolving within 1 minute), the average molecular weight of hyaluronic acid is, for example, 1000 to 8 × 10 It is preferable to set the range to about ⁴ daltons. When a pharmaceutical preparation having an adhesive property to the oral mucosa is prepared, the average molecular weight of hyaluronic acid is preferably, for example, in the range of about 1 × 10 ^{5 to} 5 × 10 ⁶ daltons. Furthermore, in the case of a pharmaceutical preparation that has an adhesive property to the oral mucosa and dissolves relatively slowly in the oral cavity (for example, dissolves within 1 minute to 20 minutes), the average molecular weight of hyaluronic acid is, for example, as follows: Preferably, it is in the range of about 3 × 10 ^{5 to} 5 × 10 ⁶ daltons. In addition, in the case of a pharmaceutical preparation used as a transdermal desensitization therapy, there is no need to adhere to the mucous membrane, and it is necessary to apply it to the skin and rapidly dissolve and release the allergen protein. Is preferably in the range of, for example, about 1000 to 8 × 10 ⁴ daltons. As the hyaluronic acid, one having a single molecular weight may be used, or one having a plurality of molecular weights may be mixed and used.

  Similarly, the pharmaceutical preparation has an adhesive property to the oral mucosa, and from the viewpoint of controlling the optimal dissolution profile for sensitizing the oral mucosa to the allergen protein more efficiently, the hyaluronic acid measured by the following measurement method is used. The intrinsic viscosity is not particularly limited, but is preferably 3 (dL / g) or more. In addition, from the viewpoint of relatively slowly dissolving the pharmaceutical preparation in the oral cavity (for example, dissolving it within 1 to 20 minutes), more preferably about 8 to 55 (dL / g), and still more preferably 8 to 40 (dL / g). (DL / g), more preferably about 10 to 40 (dL / g), and particularly preferably about 14 to 32 (dL / g). As the hyaluronic acid, one having a single molecular weight may be used, or one having a plurality of molecular weights may be mixed and used.

(Method of measuring intrinsic viscosity)
Hyaluronic acid (50 mg) was weighed and dissolved in a 0.2 mol / L sodium chloride solution to make 100 mL, and 10 mL, 15 mL, and 20 mL of this solution were weighed, and a 0.2 mol / L sodium chloride solution was added to each to 25 mL. These liquids are used as sample solutions. Viscosity measurement of each sample solution and 0.2 mol / L sodium chloride solution in accordance with the provisions of "2.53 Viscosity measurement method 1. First method Capillary viscometer method" of Japanese Pharmacopoeia 17 The specific viscosity is measured in an environment of 30.0 ± 0.1 ° C. by the method, and the reduced viscosity is calculated. A graph is drawn with the reduced viscosity on the vertical axis and the concentration of hyaluronic acid (g / 100 mL) on the horizontal axis, and the intrinsic viscosity is determined from the intersection of the straight line connecting each point and the vertical axis. In addition, hyaluronic acid used as a raw material usually contains about several% of water, but the “concentration of hyaluronic acid” in the above formula means a concentration converted to dry hyaluronic acid. . However, the specific viscosity and the reduced viscosity are obtained from the following equations.
Specific viscosity = (number of seconds of flowing down of sample solution / 0.2 mol / L second rate of flowing down of sodium chloride solution) -1
Reduced viscosity = specific viscosity / concentration of hyaluronic acid (g / 100 mL)

  The origin of hyaluronic acid is not particularly limited, and for example, those isolated and extracted from chicken cockscomb, umbilical cord and the like, and those prepared by fermentation using a microorganism such as Streptococcus or the like can be suitably used. In the present invention, commercially available products can be used as hyaluronic acid. As the hyaluronic acid, hyaluronic acid that has not been substantially chemically modified may be used, or hyaluronic acid that has been chemically modified may be used.

  Specific examples of the chemically modified hyaluronic acid include sodium carboxymethyl hyaluronate, hydroxypropyltrimonium hyaluronate, hydrolyzed alkyl hyaluronate (C12-13) glyceryl, propylene glycol hyaluronate, acetylated sodium hyaluronate and the like. Can be The chemically modified hyaluronic acid may be used alone or in combination of two or more.

  In the pharmaceutical preparation of the present invention, the ratio of hyaluronic acid in the freeze-dried body is excellent in storage stability, excellent in handling at the time of administration, and furthermore, the allergen protein is used for oral mucosa and skin. The upper limit is preferably 99.99999% by mass, 99.999% by mass, 99.99% by mass, from the viewpoint of being able to control the dissolution profile to the optimum for sensitization. 99% by mass or less, 99.9% by mass or less, and the lower limit is preferably 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass. % Or more, 80% or more, 90% or more, 95% or more, 99% or more, 99.9% or more, and the like. In the pharmaceutical preparation of the present invention, the lyophilized product may be substantially composed of only hyaluronic acid and a trace amount of allergen protein.In such a lyophilized product, for example, the content of hyaluronic acid is reduced. , 70% by weight or more, 90% by weight or more, 99% by weight or more, further 99.9% by weight or more, and the content of an allergen protein described later is, for example, 30% by weight or less, 10% by weight or less, 1% by weight. Hereinafter, it can be further set to 0.1% by mass or less.

When the molecular weight of the hyaluronic acid in the freeze-dried product is, for example, 1 × 10 ⁵ dalton or more, or when the limiting viscosity of the hyaluronic acid is 3 (dL / g) or more, handling of the pharmaceutical preparation of the present invention is performed. It has good properties and can exhibit excellent sticking properties to stick to the oral mucosa. Further, if it is 3 × 10 ⁵ daltons or more, or if the intrinsic viscosity of the hyaluronic acid is 8 (dL / g) or more, it shows elasticity by containing a small amount of water or saliva after application, so that it is necessary. During a short administration time, the pharmaceutical preparation can appropriately remain at the application site. That is, when the pharmaceutical preparation is applied to the oral mucosa, the pharmaceutical preparation has a sufficient shape-retaining property and is excellent in handleability. Further, when the pharmaceutical preparation is applied to the oral mucosa, the freeze-dried body of the pharmaceutical preparation instantaneously absorbs a small amount of water or saliva in the oral cavity and adheres to the oral mucosa. In addition, after sticking to the oral mucosa, a good elastic gel-like substance containing water and saliva is formed in a short time, so that the allergen protein can be efficiently absorbed into the oral mucosa. Further, in the case of a general pharmaceutical preparation having no sticking property to the oral mucosa, the application site in the oral cavity is limited to the sublingual, but the pharmaceutical preparation of the present invention does not limit the administration site to the sublingual mucosa. It becomes possible in any part of. Furthermore, since this gel-like material does not have thixotropic properties, damage due to physical movement in the oral cavity such as under the tongue, it is difficult to disintegrate, and it can also reduce the dissolution rate for water and saliva, Since the dissolution profile can be controlled to an optimal dissolution profile for efficiently sensitizing the oral mucosa to the allergen protein, the allergen protein can be suitably administered in the oral cavity.

In the pharmaceutical preparation of the present invention, the content per unit area of hyaluronic acid contained in the lyophilized product is not particularly limited, while exhibiting excellent storage stability, and also excellent in handleability during administration. Furthermore, it is possible to control the dissolution profile to optimally sensitize the allergen protein to the oral mucosa and skin, and for example, from the viewpoint of being suitably used for desensitization therapy, for example, 0.1 mg / cm ² or more, preferably 0 mg / cm ² or more. .5~100mg / cm ^2, more preferably about 1 to 50 mg / cm ^2, and more preferably about 1-25 mg / cm ² or so, still more preferably 2 to 20 mg / cm ² or so, particularly preferably 2 to 10 mg / cm ² .

  In the pharmaceutical preparation of the present invention, the lyophilized product may contain a polymer other than hyaluronic acid (a polymer different from hyaluronic acid). When the lyophilized product contains other polymers, it exhibits excellent storage stability and is easy to handle at the time of administration.Moreover, it is optimal for sensitizing allergen proteins to oral mucosa and skin Dissolution profile, for example, from the viewpoint of being suitably used for desensitization therapy, the content thereof is preferably 100 parts by mass of hyaluronic acid, and the upper limit is preferably 300 parts by mass or less, 200 parts by mass. Hereinafter, the amount is 100 parts by mass or less, and the lower limit is preferably 0 parts by mass or more, 20 parts by mass or more, and 40 parts by mass or more.

  In the pharmaceutical preparation of the present invention, the ratio of the total polymer in the freeze-dried body (the total ratio of hyaluronic acid and other polymers blended as required) is administered while exhibiting excellent storage stability. In addition, it is excellent in handling properties, and can be controlled to an optimal dissolution profile for sensitizing the allergen protein to the oral mucosa and skin. For example, from the viewpoint of being suitably used for desensitization therapy, 99.999999 mass % Or less, 99.999% by mass or less, 99.99% by mass or less, 99.9% by mass or less, and the lower limit is preferably 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, 95% by mass or more, 99% by mass or more, 99.9% by mass or more.

  Other polymers are not particularly limited, but exhibit excellent storage stability, are excellent in handling at the time of administration, and are most suitable for sensitizing allergen proteins to oral mucosa and skin. Dissolution profile can be controlled, for example, from the viewpoint of suitably used for desensitization therapy, preferably carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, cellulose nanofiber, crystalline cellulose, polyacrylic acid, polyvinylpyrrolidone, Polyvinyl alcohol, carrageenan, pullulan, gelatin, collagen, pectin, agarose, alginic acid, divalent alginates such as calcium, gum arabic, tragacanth gum, guar gum, chitosan, chitin derivatives , Chitosan derivatives, chondroitin sulfate, proteoglycans and the like. Among these, carboxymethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, pullulan, carrageenan, gelatin, and pectin are more preferable as other polymers. Other polymers may be used alone or in combination of two or more.

  As the allergen protein, allergens derived from pollen of trees, allergens derived from pollen of plants and plants, allergens derived from insects, allergens derived from fungi, allergens derived from animal hair, allergen proteins derived from house dust, food-derived allergens Allergens and the like can be mentioned, and there is no particular limitation as long as the antigen specifically reacts with an antibody of a person having an allergic disease, and a known allergen protein can be used. Specific examples of allergen proteins include ovalbumin, casein, soy protein, wheat protein, and other food allergen proteins, cedar, harugaya, kamogaya, hosomugi, ooseagaeri, Nagahagusa, ragweed, and the like used in desensitization therapy. And mite allergen proteins such as Dermatophagoides farinae and Dermatophagoides pteronyssinus. One type of allergen protein may be used alone, or two or more types may be mixed and used.

  Specific examples of the pollen allergen protein include acacia, alder, velvet blue daimo, rape birch, birch, maple, mountain cedar, red cedar, cottonwood, cypress, American elm, akinil, togasawara, rubber tree, eucalyptus tree, enoki, Hickory, American linden, sugar maple, mesquite, cassia, oak, olive, pecan, pepper, pine, botany, russian olives, american moss, pollen allergen proteins of trees such as iwasushi, black walnut, black willow; Sparrow blossoms, corn, broadleaf foxglove, sorghum sorghum, oats, duckweed, konukagusa, hosomugi, rice, harugaya, giant irises, cypress, akaza, onamimi, sagegrass, seitakawa Goldenrod, Isohouki, common lambsquarters, calendula, nettle, Aobie, ribwort, Ambrosia trifida, ragweed, ragweed beetle, Noharahijiki, sagebrush, broom, and a pollen allergen protein plants such as Rumex acetosella. For example, allergen proteins derived from cedar pollen include Cry j1, Cry j2, Cyr j3, and the like.

  Examples of mite allergen proteins include, for example, allergens derived from feces of Dermatophagoides pteronyssinus (Der p1), allergens derived from insect bodies (Der p2), and allergens derived from Dermatophagoides federae from Dermatophagoides federae. And insect-derived allergens (Der f2).

In the pharmaceutical preparation of the present invention, the ratio of the allergen protein in the freeze-dried body may be appropriately adjusted so as to be the content required for the pharmaceutical preparation, while exhibiting excellent storage stability, and It is also excellent in handleability, and can be controlled to an optimal dissolution profile for sensitizing the allergen protein to the oral mucosa and skin, and from the viewpoint of being suitably used for desensitization therapy, the upper limit is preferably 50 % By mass, 1% by mass or less, 0.1% by mass or less, and the lower limit is preferably 1 × 10 ⁻¹⁰ % by mass or more and 1 × 10 ⁻⁸ % by mass or more and 1 × 10 ⁻⁷ % by mass. % By mass or more.

Further, in the pharmaceutical preparation of the present invention, the content per unit area of the allergen protein contained in the lyophilized product may be appropriately adjusted so as to be the content required for the pharmaceutical preparation, and excellent storage stability is obtained. While exerting, it is also excellent in handling at the time of administration, furthermore, it can be controlled to the optimal dissolution profile for sensitizing allergen proteins to oral mucosa and skin, and from the viewpoint of being suitably used for desensitization therapy For example, 0.01 ng / cm ² or more, preferably about 0.1 ng / cm ^{2 to} 10 mg / cm ² , more preferably about 1 ng / cm ^{2 to 5} mg / cm ² , and still more preferably 10 ng / cm ^{2 to 1} mg / cm. ^{About 2} , more preferably about 50 ng / cm ^{2 to} 500 μg / cm ² , particularly preferably about 100 ng / cm ^{2 to} 100 μg / cm ^2. No.

  The freeze-dried body may further contain other components in addition to the hyaluronic acid, the allergen protein, and a polymer different from the hyaluronic acid to be added as required. Other components are not particularly limited as long as the effects of the present invention are not impaired, and include, for example, sugars, polyhydric alcohols, sugar fatty acids, pH adjusters and the like.

  By containing any one of sugar, polyhydric alcohol, and sugar fatty acid ester in the freeze-dried product, the stiffness of the freeze-dried product is increased, and the handleability of the pharmaceutical preparation is further improved. In addition, since the dissolution time in the oral cavity is delayed, the pharmaceutical preparation can be preferably left on the oral mucosa.

  Sugars include, for example, monosaccharides, disaccharides, tri-hexasaccharides, maltodextrins and their sugar alcohols. Examples of the monosaccharides include aldotetroses such as erythrose and threose, ribose, lyxose, xylose, aldpentose such as arabinose, allose, talose, growth, aldohexose such as glucose, altrose, mannose, galactose, and idose; Keto pentoses such as ketotetroses such as erythrulose, xylulose and ribulose; ketohexoses such as psicose, fructose, sorbose and tagatose. Examples of the disaccharide include α-diglucosides such as trehalose, kojibiose, nigerose, maltose, and isomaltose; β-diglucosides such as isotrehalose, sophorose, laminaribiose, cellobiose, and gentiobiose; and α, β-diglucosides such as neotrehalose. And lactose, sucrose, isomaltulose (palatinose), sucralose and the like. Examples of the trisaccharide include raffinose and the like. Examples of the trisaccharide to hexasaccharide oligosaccharides include, for example, cyclic oligosaccharides such as fructooligosaccharides, galactooligosaccharides, xylo-oligosaccharides, isomaltooligosaccharides, chitin oligosaccharides, chitosan oligosaccharides, oligoglucosamines, dexulolines, and cyclodextrins. . Examples of the monosaccharide alcohol include, for example, tetritol such as erythritol, D-threitol and L-threitol, pentitol such as D-arabinitol and xylitol, D-idinol, galactitol (darcitol), and D-glucitol (sorbitol). And hexitols such as mannitol and cyclitols such as inositol. Examples of the disaccharide alcohol include maltitol, lactitol, and reduced palatinose (isomalto), and examples of the oligosaccharide include pentaerythritol and reduced maltose syrup. Among these, maltose, sorbitol, mannitol, sucrose, raffinose, trehalose, xylitol, erythol, isomalt, and maltodextrin are particularly preferred. One type of sugar may be used alone, or two or more types may be used in combination. In the present invention, the term “sugar” does not include a liquid polyhydric alcohol at 25 ° C., and the liquid polyhydric alcohol is included in the “polyhydric alcohol” described later.

  The polyhydric alcohol is not particularly limited, but is preferably glycerin such as glycerin or diglycerin; propylene glycol such as propylene glycol or dipropylene glycol; 1,3-propanediol, butanediol (1,3- Butanediol, 1,4-butanediol, etc.), 1,2-pentanediol, 1,2-hexanediol, 1,2-octanediol; ethylene glycols such as ethylene glycol, diethylene glycol, triethylene glycol, and polyethylene glycol And more preferably glycerin, diglycerin, propylene glycol, 1,3-butanediol, polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, polyethylene glycol. Call 1000, polyethylene glycol 1500, polyethylene glycol 6000, polyethylene glycol 20000,1,3- propanediol, 1,2-pentanediol, 1,2-hexanediol and the like. Among these, glycerin, diglycerin, polyethylene glycol having a molecular weight of less than 1,000 are particularly preferable. The polyhydric alcohol may be used alone or in combination of two or more.

  Examples of sugar fatty acids include sorbitan fatty acid esters and sucrose fatty acid esters. Examples of the sorbitan fatty acid ester include sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, sorbitan coconut oil fatty acid, and polyoxyethylene sorbitan fatty acid ester. Examples of the sucrose fatty acid ester include sucrose stearate, sucrose oleate, sucrose palmitate, sucrose myristate, sucrose behenate, sucrose erucate, and sucrose. Mixed fatty acid esters and the like can be mentioned. The sugar fatty acids may be used alone or in combination of two or more.

  As the content of sugar, polyhydric alcohol, or sugar fatty acid in the freeze-dried body (total content when plural types are included), it is easy to handle at the time of administration while exhibiting excellent storage stability In addition, it is possible to control the optimal dissolution profile for sensitizing the allergen protein to the oral mucosa and the skin. For example, from the viewpoint of suitably used for desensitization therapy, the upper limit is preferably 80 Mass% or less, 70 mass% or less, 60 mass% or less, 50 mass% or less, 30 mass% or less, 20 mass% or less, 10 mass% or less, 5 mass% or less, 0 mass%. Preferably, 0 mass% or more, 5 mass% or more, 10 mass% or more, 15 mass% or more are mentioned.

  In the production process of the pharmaceutical preparation of the present invention, it is preferable that the pH of an aqueous solution for a pharmaceutical preparation described below immediately before freeze-drying is 5.0 to 9.0. When the pH is in the above range, it is possible to prevent the physicochemical stability of the allergen protein from being remarkably reduced, and to ensure safety. More preferably, the pH is 6.0 to 8.0.

  In order to adjust the pH of the aqueous solution for a pharmaceutical preparation, a pH adjuster may be contained as necessary. The pH adjuster is not particularly limited, but may be adipic acid, aqueous ammonia, hydrochloric acid, sodium carbonate, dilute hydrochloric acid, citric acid hydrate, glycine, glucono-δ-lactone, gluconic acid, crystalline sodium dihydrogen phosphate, succinic acid Acetic acid, ammonium acetate, sodium acetate hydrate, diisopropanolamine, tartaric acid, potassium hydroxide, calcium hydroxide, sodium hydroxide, magnesium hydroxide, sodium bicarbonate, sodium carbonate hydrate, triisopropanolamine, triethanol Amine, carbon dioxide, lactic acid, sodium lactate solution, glacial acetic acid, fumaric acid, monosodium fumarate, sodium propionate, boric acid, ammonium borate, borax, maleic acid, anhydrous citric acid, anhydrous sodium monohydrogen phosphate, Anhydrous sodium dihydrogen phosphate, Megur Methanesulfonic acid, monoethanolamine, sulfuric acid, aluminum potassium sulfate hydrate, DL-malic acid, phosphoric acid, trisodium phosphate, dipotassium phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, etc. No. These pH adjusters may be used alone or in a combination of two or more. The pH adjuster is, for example, a combination of an organic acid and an organic acid salt equivalent to an organic acid salt described later, such as a combination of an organic acid and sodium chloride, calcium chloride, magnesium chloride, or potassium chloride. You may. From the viewpoint of production, it is preferable that the pH adjuster can adjust the pH with a small amount. Examples of such a pH adjuster include hydrochloric acid and sodium hydroxide. Further, as the above-mentioned pH adjuster, an organic acid and an organic acid salt having an effect of suppressing denaturation of an enzyme are also suitably used. Such organic acids include, for example, citric acid hydrate, glycine, glucono-δ-lactone, gluconic acid, succinic acid, acetic acid, sodium acetate hydrate, tartaric acid, lactic acid, glacial acetic acid, fumaric acid, fumaric acid Monosodium, sodium propionate, maleic acid, citric anhydride, malic acid and the like. Further, organic acid salts include, for example, amino acid salts, adipates, citrates, malates, acetates, succinates, propionates, butyrates, malonates, glutarates, maleates, Glycolate, lactate, gluconate, fumarate, tartrate, glycyrrhizinate, pimelate, aspartate and the like can be mentioned. These organic acid salts may be used alone or in a combination of two or more.

  The pharmaceutical preparation of the present invention is excellent in storage stability, and further has a molecular weight and a content of hyaluronic acid, and other components (for example, the above-mentioned other polymer, sugar, polyhydric alcohol, By adjusting the type and content of sugar fatty acid esters, pH adjusters and the like), the pharmaceutical preparation of the present invention can contain an active ingredient other than the allergen protein. If so, the active ingredient can be released in the oral cavity for a long period of time.

  Further, the pharmaceutical preparation of the present invention may contain, as components constituting the freeze-dried product, in addition to the above-mentioned components, flavors, flavoring agents, sweetening agents, coloring agents, preservatives, antioxidants, and stabilizers as necessary. , An antifoaming agent, a surfactant and the like may be appropriately used.

  In the present invention, the thickness of the freeze-dried body is not particularly limited, but preferably about 0.05 to 10 mm when the pharmaceutical preparation is constituted by a single layer of the freeze-dried body. When the pharmaceutical preparation is a laminate of a freeze-dried product and a base material described later, the thickness of the freeze-dried product is preferably about 0.02 to 10 mm. In addition, the freeze-dried body prepared by freeze-drying can be thinned by compression to obtain a pharmaceutical preparation.

In the present invention, the area of the freeze-dried body can also be appropriately set according to the size of the application site (for example, below the tongue), and for example, about 0.5 to ¹⁰ cm ² . Furthermore, the shape of the freeze-dried body is not particularly limited, and examples thereof include a shape under the tongue, a circle, an ellipse, a polygon (such as a triangle, a square, a pentagon, and a hexagon), a star, and an irregular shape. Can be

  The pharmaceutical preparation 10 of the present invention may be constituted by a single layer of a freeze-dried body 1 as shown in FIG. 1, for example, or as shown in FIG. And the freeze-dried body 1 laminated on the second layer 2. 1 and 2, the surface of the freeze-dried body 1 constitutes an adhesive surface 10a with the oral mucosa or the skin.

  By providing the pharmaceutical preparation of the present invention with a base material, an effect of further improving the handleability of the pharmaceutical preparation can be expected. In addition, it is possible to prevent the pharmaceutical preparation from being damaged or disintegrated due to physical movements in the oral cavity such as the sublingual part, and to reduce the dissolution rate of the pharmaceutical preparation in water or saliva in the oral cavity. The effect of controlling the optimal dissolution profile for sensitizing the allergen protein can be expected. For example, by using a base material having high mechanical strength and high water resistance as a material constituting the base material, and having poor adhesion to a wet hand or the like than a freeze-dried body containing hyaluronic acid, such an effect can be obtained. It is possible to exhibit it suitably.

  The material constituting the base material is not particularly limited, but from the viewpoint of exhibiting such effects, preferably, the above-mentioned other polymer, or a polymer obtained by adding a polyhydric alcohol to another polymer. For example, by using a freeze-dried body of the other polymer described above as a base material, such an effect can be suitably exerted. As a material constituting the base material, particularly preferably, a lyophilized product of an aqueous solution containing another polymer such as divalent alginate such as methylcellulose, hydroxypropylmethylcellulose, cellulose nanofiber, agarose, and calcium, and a polymer and molecular weight of these polymers Lyophilized product of an aqueous solution containing 1,000 or more polyethylene glycols. Other polymers may be used alone or in combination of two or more, including hyaluronic acid.

  Further, by using a thin film composed of hyaluronic acid and the other polymer described above as a base material, such an effect can be suitably exerted. Examples of the material constituting the base material include hyaluronic acid, the above-mentioned other polymers, and biodegradable polymers such as polylactic acid and other polymers to which a plasticizer such as a polyhydric alcohol is added. Hyaluronic acid and the plasticizer to be added to the other polymer may be used alone or in combination of two or more.

  When the pharmaceutical preparation of the present invention comprises a substrate, the thickness of the substrate is not particularly limited, but preferably 0.0001 to 10 mm, more preferably 0.0002 to 5 mm, or 0.01 to 10 mm. .

  The application site of the pharmaceutical preparation of the present invention includes, for example, the surface of the oral mucosa such as the sublingual surface. In addition, eating and drinking may be performed in a state where the pharmaceutical preparation of the present invention is adhered to the oral mucosa.

  Another application site of the pharmaceutical preparation of the present invention includes the skin surface as in the case of the transdermal absorption preparation.

  When the pharmaceutical preparation of the present invention is applied to the skin, a drug integrated with an adhesive tape or the like using a thin polymer film (polyolefin such as polyurethane or polyethylene, film such as vinyl chloride or polyethylene terephthalate) as a support is defined as a drug. Alternatively, the pharmaceutical preparation of the present invention may be used as a pharmaceutical, and the pharmaceutical preparation of the present invention may be applied to a thin polymer film (polyolefin such as polyurethane or polyethylene, a film of vinyl chloride, polyethylene terephthalate or the like) as an adhesive tape or the like. It may be fixed and used.

  In addition, when the pharmaceutical preparation of the present invention is applied to the skin, the outermost layer of the skin becomes a barrier, and the absorption of allergen proteins into the skin is easily hindered. , A tape stripping technique, a microneedle technique, or the like.

  The method for producing the pharmaceutical preparation of the present invention is not particularly limited. For example, a method including a step of preparing a composition containing at least an allergen protein, hyaluronic acid, and water, and a step of freeze-drying the composition is provided. No.

  In the method for producing a pharmaceutical preparation of the present invention, details of hyaluronic acid, allergen protein, other polymers blended as required, and the content of each component are as described above. In addition, the contents of hyaluronic acid, allergen protein, and other polymers to be blended as necessary may be adjusted so as to become these contents in the above-mentioned pharmaceutical preparation.

  When the pharmaceutical preparation of the present invention is constituted by a multilayer comprising a base material and a freeze-dried body laminated on the base material, for example, the base material is formed of another polymer as described above. In the case of being constituted by a freeze-dried product of the above, one of the composition containing another polymer and water, and one of the composition containing hyaluronic acid, allergen protein and water is frozen, and the freezing is placed in a container. A method of pour-freezing the other composition onto the body and freeze-drying the two can produce a multi-layered pharmaceutical preparation. Further, for example, when the substrate is formed of a film of another polymer as described above, a composition containing hyaluronic acid, an allergen protein, and water was poured into a container and frozen by placing the film on the container. Thereafter, a method of freeze-drying may be mentioned.

  Further, for example, a composition containing hyaluronic acid, an allergen protein and water is laminated on a substrate placed in a container, and the composition is freeze-dried in that state to produce a multi-layered pharmaceutical preparation. can do.

  Hereinafter, the present invention will be described in detail with reference to Examples and Comparative Examples. However, the present invention is not limited to the examples. The reagents used in the following Examples and Comparative Examples are as follows.

<Reagent>
-Hyaluronic acid (2.3 million): Sodium hyaluronate, trade name "HYALURONSAN HA-LQSH" manufactured by Kewpie Co., Ltd. (Product designation: molecular weight 1.6 million to 2.9 million, average molecular weight 2.3 million, intrinsic viscosity 25.0 to 40.0 dL) / G)
-Hyaluronic acid (1.2 million): sodium hyaluronate, trade name "Hyaluronsan HA-LQ" manufactured by Kewpie Co., Ltd. (Product notation: molecular weight: 85 to 1.6 million; average molecular weight: 1.2 million, intrinsic viscosity: 15.0 to 25.0 dL / g)
-Hyaluronic acid (350,000): Sodium hyaluronate, trade name "HA-LF-P" manufactured by Kewpie Co., Ltd. (Product designation: 200,000 to 500,000 molecular weight, average molecular weight 350,000) (used in Examples) Intrinsic viscosity of reagent 8.7dL / g)
-Hyaluronic acid (100,000): Sodium hyaluronate, trade name "Hyaluronic acid FCH-SU" manufactured by Kikkoman Biochemifa Co., Ltd. (Product display: average molecular weight 50,000 to 110,000)
-Hyaluronic acid (50,000 or less): Hyaluronic acid, a product name of Hypeuronsan HA-LF5-A manufactured by Kewpie Co., Ltd. (Product display: molecular weight of 50,000 or less, average molecular weight of 35,000)
-Hyaluronic acid (2000): Hyaluronic acid, trade name "Hyaronano" manufactured by Kewpie Co., Ltd. (Product display: average molecular weight 2000)
・ Carboxymethylcellulose: sodium carboxycellulose, trade name “Selogen F-BSH-12” manufactured by Daiichi Kogyo Seiyaku Co., Ltd.
・ Hydroxypropylcellulose: trade name “Hydroxypropylcellulose H” manufactured by Nippon Soda Co., Ltd.
・ Polyvinylpyrrolidone: trade name “Polyvinylpyrrolidone K90 (for molecular biology)” manufactured by Wako Pure Chemical Industries, Ltd.
-Pullulan: Pullulan manufactured by Wako Pure Chemical Industries, Ltd. (for biochemistry)
-Sorbitol: D (+)-sorbitol (first grade) manufactured by Wako Pure Chemical Industries, Ltd.
・ Trehalose: Trehalose dihydrate (special grade) manufactured by Wako Pure Chemical Industries, Ltd.
-Mannitol: D (-)-mannitol (special grade) manufactured by Wako Pure Chemical Industries, Ltd.
・ Sucrose: Sucrose (special grade) manufactured by Wako Pure Chemical Industries, Ltd.
・ Glycerin: Glycerin (special grade) manufactured by Wako Pure Chemical Industries, Ltd.
・ Polyethylene glycol 1000: Polyethylene glycol 1000 (lump) manufactured by Wako Pure Chemical Industries, Ltd.
-Cedar pollen extract: "cedar pollen extract" manufactured by ITEA Corporation (cedar pollen allergen Cry j1 concentration 10 µg / mL)
-Mite extract: "Mite Extract-Df" manufactured by LSL Co., Ltd. (Gypsophilia allergen concentration Der f1 23 μg / mL, Der f2 87 μg / mL)
-Egg albumin: Sigma's Album From Chicken Egg White, Grade V
・ Sodium hydroxide: sodium hydroxide manufactured by Wako Pure Chemical Industries, Ltd. (special grade)

[Examples 1 to 21]
(Manufacture of pharmaceutical preparations)
Aqueous solutions for pharmaceutical preparations (compositions containing at least hyaluronic acid, allergen proteins, and water) were prepared so as to have the compositions shown in Table 1, respectively. Specifically, first, hyaluronic acid having a molecular weight shown in Table 1 and a ginger allergen (total amount of freeze-dried ginger allergens Der f1 and Derf2) were purified water (the total amount of water is the amount of purified water described in the table). To prepare a pharmaceutical formulation aqueous solution. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation.

<Evaluation of properties of pharmaceutical preparations>
The properties of the pharmaceutical preparations obtained in Examples 1 to 21 were evaluated according to the following criteria. The thickness was measured. Table 2 shows the results.

(Properties of pharmaceutical preparations)
A: It is a cotton-like dry body and has no waist as it is soft. After storage at 25 ° C. for 2 hours, there is no property change due to moisture absorption or the like.
B: It is a cotton-like dry body, which is fluffy but has a waist. After storage at 25 ° C. for 2 hours, there is no property change due to moisture absorption or the like.
C: It is a cotton-like dried body, has little flexibility, and has a waist, but has no brittleness. After storage at 25 ° C. for 2 hours, there is no property change due to moisture absorption or the like.
D: A high-density sponge-like dried product having little flexibility but no brittleness. After storage at 25 ° C. for 2 hours, there is no property change due to moisture absorption or the like.

<Evaluation of solubility of pharmaceutical preparations>
The pharmaceutical preparations obtained in Examples 1 to 21 were placed in a container (having a volume of 1 L, a height of 160 to 210 mm, and an inner diameter of 98 to 106 mm) using an apparatus of the general test method "dissolution test" of the 17th revised Japanese Pharmacopoeia. ) Was charged with 800 mL of physiological saline, and the container was placed in a thermostatic water bath and adjusted so that the temperature in the container during the test was 37 ± 0.5 ° C. Each of the pharmaceutical preparations obtained above was gently poured into the water surface of this container, and the time required for the pharmaceutical preparation to completely dissolve was evaluated according to the following criteria. Table 2 shows the results.

(Dissolution time of pharmaceutical preparation)
A: Within 10 seconds B: Within 1 minute C: Within 5 minutes D: Within 10 minutes E: Within 20 minutes F: Over 20 minutes

<40 ° C storage stability test>
The pharmaceutical preparations obtained in Examples 1 to 21 were sealed in an aluminum pouch, and mite allergen activity after storage at 40 ° C for 1 week was measured. In addition, the properties and solubility of the pharmaceutical preparation after storage at 40 ° C. for one week were evaluated according to the method described above. Table 2 shows the results.

(Dani allergen activity measurement)
Using a mite allergen (Der f1) assay kit (manufactured by Nichinichi Pharmaceutical Co., Ltd.), the allergen activity of Der f1, one of the major allergens of mites, was measured. The pharmaceutical preparation was placed in a diluent (buffer) included in the kit and completely dissolved. Assuming that the allergen in the pharmaceutical preparation was not inactivated at all, the resulting solution was diluted to a Der f1 concentration of 10 ng / mL and prepared as a sample for measurement. Next, a standard solution (32 ng / mL, 16 ng / mL, 8 ng / mL, 4 ng / mL) or 50 μL of the sample for measurement is added to the antibody-coated plate, and 50 μL of the secondary antibody solution is further added. The reaction was performed for 2 hours. Next, 100 μL of the enzyme solution was added, and the reaction was carried out at 37 ° C. for 1 hour. Next, 100 μL of the substrate solution was added, and the reaction was carried out at 37 ° C. for 5 minutes to develop a color. Finally, 100 μL of a reaction stop solution was added. Thereafter, the ultraviolet absorption intensity at 450 nm was measured. A calibration curve was determined based on the absorption intensity of the standard solution at each Der f1 concentration, and the Der f1 allergen activity (ng / mL) of each measurement sample was measured in accordance therewith, and the pharmaceutical preparation was sealed in an aluminum pouch and stored at -32 ° C. The allergen activity of the sample stored at 40 ° C. for one week, based on the allergen activity of the sample stored for one week as 100%, was evaluated according to the following criteria.
A: 90% or more and less than 105% B: 70% or more and less than 90% C: 50% or more and less than 70% D: 30% or more and less than 50% E: less than 30%

[Examples 22 to 25]
(Manufacture of pharmaceutical preparations)
An aqueous solution for a pharmaceutical preparation (a composition containing at least hyaluronic acid, an allergen protein, and water) was prepared so as to have the compositions shown in Table 3, respectively. Specifically, first, the hyaluronic acid (molecular weight 100,000), other polymers (polymers different from hyaluronic acid), and ginger root allergens (the total amount of freeze-dried ginger root allergens Der f1 and Der f2) shown in Table 3 ) Was uniformly dissolved in purified water (the total amount of water was as shown in the table) to prepare an aqueous solution for pharmaceutical preparation. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation. Next, in the same manner as in Examples 1 to 21, a storage stability test (evaluation of allergen activity, properties, and solubility) of each pharmaceutical preparation was performed. Table 4 shows the results.

[Examples 26 to 33]
(Manufacture of pharmaceutical preparations)
Aqueous solutions for pharmaceutical preparations (compositions containing at least hyaluronic acid, allergen proteins, and water) were prepared so as to have the compositions shown in Table 5, respectively. Specifically, first, hyaluronic acid (molecular weight 100,000), sugar / polyhydric alcohol, and ginger allergen (freeze-dried total amount of ginger allergens Der f1 and Der f2) shown in Table 5 were purified water (water Was uniformly dissolved in the amount of purified water described in the table) to prepare an aqueous solution for pharmaceutical preparation. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation. Next, in the same manner as in Examples 1 to 21, a storage stability test (evaluation of allergen activity, properties, and solubility) of each pharmaceutical preparation was performed. Table 6 shows the results.

[Examples 34 to 52]
(Manufacture of pharmaceutical preparations)
Aqueous solutions for pharmaceutical preparations (compositions containing at least hyaluronic acid, allergen proteins, and water) were prepared so as to have the compositions shown in Table 7, respectively. Specifically, first, hyaluronic acid and cedar pollen allergen having the respective molecular weights shown in Table 7 (the amount of freeze-dried Cry j1) are uniformly dissolved in purified water (the total amount of water is the purified water amount shown in the table). Thus, an aqueous solution for a pharmaceutical formulation was prepared. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation. Next, in the same manner as in Examples 1 to 21, a storage stability test (evaluation of allergen activity, properties, and solubility) of each pharmaceutical preparation was performed. The cedar pollen allergen activity was evaluated according to the method described below. Table 8 shows the results.

(Measurement of cedar pollen activity)
Using a cedar pollen allergen (Cry j1) assay kit (manufactured by Shibayagi Co., Ltd.), the allergen activity of Cry j1, which is one of the main allergens of cedar, was measured. The allergen-containing preparation was put in the buffer solution provided with the kit and completely dissolved. Assuming that the allergen in the preparation was not inactivated at all, it was diluted so that the concentration of Cry j1 in the obtained solution was 5 ng / mL, and prepared as a sample for measurement. Next, a standard solution (10 ng / mL, 5 ng / mL, 2.5 ng / mL, 1.25 ng / mL) or 100 μL of the measurement sample was added to the antibody-coated plate, and reacted at room temperature for 1 hour. Next, 100 μL of an HRP-conjugated anti-Cry j1 antibody solution was added, and the reaction was carried out at room temperature for 1 hour. Next, 100 µL of a coloring solution was added, and the reaction was carried out at room temperature for 20 minutes to develop a color. Finally, 100 µL of a reaction stopping solution was added. Thereafter, the ultraviolet absorption intensity at 450 nm was measured. A calibration curve was determined based on the absorption intensity of the standard solution at each concentration of Cry j1, the Cry j1 allergen activity (ng / mL) of each measurement sample was measured in accordance with the calibration curve, and the pharmaceutical preparation was sealed in an aluminum pouch and stored at -32 ° C. The allergen activity of the sample stored at 40 ° C. for one week, based on the allergen activity of the sample stored for one week as 100%, was evaluated according to the following criteria.
A: 90% or more and less than 105% B: 70% or more and less than 90% C: 50% or more and less than 70% D: 30% or more and less than 50% E: less than 30%

[Examples 55 to 58]
(Manufacture of pharmaceutical preparations)
Aqueous solutions for pharmaceutical preparations (compositions containing at least hyaluronic acid, allergen proteins, and water) were prepared so as to have the compositions shown in Table 9, respectively. Specifically, first, hyaluronic acid (molecular weight 100,000), another polymer (a polymer different from hyaluronic acid), and cedar pollen allergen (freeze-dried amount of Cry j1) shown in Table 9 were purified water (water Was uniformly dissolved in the amount of purified water described in the table) to prepare an aqueous solution for pharmaceutical preparation. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation. Next, a storage stability test (evaluation of allergen activity, properties, and solubility) of each of the pharmaceutical preparations was performed in the same manner as in Examples 34 to 54. Table 10 shows the results.

[Examples 59 to 66]
(Manufacture of pharmaceutical preparations)
Aqueous solutions for pharmaceutical preparations (compositions containing at least hyaluronic acid, allergen proteins, and water) were prepared so as to have the compositions shown in Table 11, respectively. Specifically, first, hyaluronic acid (molecular weight 100,000), sugar / polyhydric alcohol, and cedar pollen allergen (freeze-dried amount of Cry j1) described in Table 11 were purified water (the total amount of water was described in the table). Of purified water) to prepare an aqueous solution for pharmaceutical preparation. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation. Next, in the same manner as in Examples 34 to 54, the properties, solubility, and storage stability at 40 ° C of each pharmaceutical preparation were evaluated. Table 12 shows the results.

[Reference Examples 1 to 6]
(Manufacture of placebo formulation)
Hyaluronic acid having each molecular weight shown in Table 13 was uniformly dissolved in purified water (the total amount of water was purified water shown in the table) to prepare an allergen-free aqueous solution of hyaluronic acid. Next, the obtained aqueous solution of hyaluronic acid was placed in a plastic tray of 2 × 2.5 cm (5 cm ² ) and frozen in a freezer at −82 ° C. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product of hyaluronic acid. The obtained freeze-dried product was used as a placebo preparation.

(Volunteer evaluation)
The placebo preparations prepared in Reference Examples 1 to 6 were applied under the tongue of a human volunteer whose oral cavity was rinsed and cleaned with 100 ml of distilled water. After standing for 10 minutes as it was, evaluation was made according to the following criteria. Table 14 shows the results.

(Volunteer evaluation criteria)
A: The placebo formulation dissolved within 10 seconds after sublingual application.
B: After application under the tongue, it adhered to the mucous membrane and the placebo formulation dissolved within 1 minute.
C: After application under the tongue, it adhered to the mucous membrane and the placebo preparation dissolved within 10 minutes.
D: After application under the tongue, it adhered to the mucous membrane, and the placebo preparation remained under the tongue even after 10 minutes.

[Examples 67 to 70]
(Manufacture of pharmaceutical preparations)
Aqueous solutions for pharmaceutical preparations (compositions containing at least hyaluronic acid, allergen proteins, and water) were prepared so as to have the compositions shown in Table 15, respectively. Specifically, first, the hyaluronic acid described in Table 15 is uniformly dissolved in a half amount of purified water described in the table, and an appropriate amount of 1% sodium hydroxide is uniformly dissolved. The purified water was added so as to obtain the purified water amount to obtain a hyaluronic acid aqueous solution having a pH of 6.3. This pH was measured using TwinpH (B-212) manufactured by Horiba, Ltd. Egg albumin and trehalose were uniformly dissolved in this aqueous solution of hyaluronic acid to prepare an aqueous solution for pharmaceutical preparation. Next, the obtained aqueous solution for pharmaceutical preparation was prepared. Next, the obtained aqueous solution for pharmaceutical preparations was placed in a 2 × 2.5 cm (5 cm ² ) plastic tray and frozen in a −82 ° C. freezer. Next, the aqueous solution for pharmaceutical preparations was lyophilized (24 hours) in a tray at a pressure of 10 Pa or less to obtain a lyophilized product. The obtained freeze-dried product was used as a pharmaceutical preparation. Next, a storage stability test (evaluation of properties and solubility) of each pharmaceutical preparation was performed at 40 ° C in the same manner as in Examples 1 to 21. Table 16 shows the results.

(Evaluation of application of pharmaceutical preparation to rat skin)
Under anesthesia by intraperitoneal administration of a urethane solution (1.6 g / kg), the abdomen of an 8-week-old Wistar male rat was dehaired, and the dehaired portion was white cloth adhesive plaster (Nichiban H25) manufactured by Nichiban Co., Ltd. After complete exfoliation using, the pharmaceutical preparation prepared in Example 68 was applied to the removed site, and immediately HYPERLINK "http://www.mmm.co.jp/corporate/group/japan.html" The pharmaceutical preparation was coated with 3M Tegaderm Transparent Dressing manufactured by Japan Co., Ltd. As a result, the pharmaceutical preparation was completely dissolved within 1 minute after application to the skin.

DESCRIPTION OF SYMBOLS 1 Lyophilized body 2 Substrate 10 Pharmaceutical preparation 10a Adhesion surface with oral mucosa

Claims (12)

  A pharmaceutical preparation comprising a lyophilized composition containing at least an allergen protein and hyaluronic acid. The molecular weight of the hyaluronic acid is 1 × 10 ⁵ daltons or pharmaceutical formulation according to claim 1. The pharmaceutical preparation according to claim 1, wherein the molecular weight of the hyaluronic acid is 3 × 10 ⁵ dalton or more. The pharmaceutical preparation according to any one of claims 1 to 3, wherein the intrinsic viscosity of the hyaluronic acid measured by the following measurement method is 3 dL / g or more.
(Method of measuring intrinsic viscosity)
Hyaluronic acid (50 mg) was weighed and dissolved in a 0.2 mol / L sodium chloride solution to make 100 mL, and 10 mL, 15 mL, and 20 mL of this solution were weighed, and a 0.2 mol / L sodium chloride solution was added to each to 25 mL. These liquids are used as sample solutions. Viscosity measurement of each sample solution and 0.2 mol / L sodium chloride solution in accordance with the provisions of "2.53 Viscosity measurement method 1. First method Capillary viscometer method" of Japanese Pharmacopoeia 17 The specific viscosity is measured in an environment of 30.0 ± 0.1 ° C. by the method, and the reduced viscosity is calculated. A graph is drawn with the reduced viscosity on the vertical axis and the concentration of hyaluronic acid (g / 100 mL) on the horizontal axis, and the intrinsic viscosity is determined from the intersection of the straight line connecting each point and the vertical axis.
However, the specific viscosity and the reduced viscosity are obtained from the following equations.
Specific viscosity = (number of seconds of flowing down of sample solution / 0.2 mol / L second rate of flowing down of sodium chloride solution) -1
Reduced viscosity = specific viscosity / concentration of hyaluronic acid (g / 100 mL)   The pharmaceutical preparation according to claim 4, wherein the intrinsic viscosity of the hyaluronic acid measured by the above measurement method is 8 dL / g or more.   The pharmaceutical preparation according to any one of claims 1 to 5, wherein the allergen protein includes at least one of a pollen allergen protein and a mite allergen protein.   The pharmaceutical preparation according to any one of claims 1 to 6, comprising a substrate and the lyophilized product laminated on the substrate.   The pharmaceutical preparation according to any one of claims 1 to 7, wherein the freeze-dried body contains a polymer different from hyaluronic acid.   The pharmaceutical preparation according to any one of claims 1 to 8, wherein the freeze-dried product further contains at least one of sugar, polyhydric alcohol, and sugar fatty acid.   The pharmaceutical preparation according to any one of claims 1 to 9, which is used for sublingual administration.   The pharmaceutical preparation according to any one of claims 1 to 9, which is used for transdermal administration. A method for producing a pharmaceutical preparation according to any one of claims 1 to 11,
Preparing a composition comprising at least an allergen protein, hyaluronic acid, and water,
Freeze-drying the composition;
A method for producing a pharmaceutical preparation, comprising:

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