JPH08225447A - Hyaluronic acid decomposition inhibitor and therapeutic agent for disease involved in hyaluronic acid decomposition - Google Patents

Abstract

PURPOSE: To provide an inhibitor inhibiting hyaluronic acid decomposition through acting on the cells present in human connective tissue, and to provide a therapeutic agent containing this inhibitor. CONSTITUTION: This hyaluronic acid decomposition inhibitor containing, as active ingredient, a histamine H1 -antagonist (salt) such as an ethanolamine-type diphenhydramine, ethylenediamine-type pyrilamine or tripelennamine, alkylamine- type chlorpheniramine or brompheniramine, piperazine-type cyclizine, or phenothiazine-type promethazine. The therapeutic agent for diseases involved in hyaluronic acid decomposition contains this inhibitor.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a hyaluronic acid degradation inhibitor which acts on cells existing in human connective tissue and inhibits hyaluronic acid degradation, and further relates to a therapeutic agent for diseases associated with the degradation of hyaluronic acid.

[0002]

2. Description of the Related Art Conventionally, histamine H ₁ -antagonists antagonize histamine released from mast cells at the time of inflammation and exert their action on vascular endothelial cells and smooth muscle cells.
It is known as a major anti-inflammatory drug because it suppresses binding to the ₁ -receptor. It is also known to act on the central nervous system due to its anticholinergic effect. However, from the above mechanism of action, at present, pollinosis, urticaria, rhinitis, conjunctivitis, atopic skin inflammation,
Contact dermatitis, edema, itching, allergic disease or motion sickness,
It is only used as a therapeutic agent and a hypnotic drug.

On the other hand, hyaluronic acid retains water in the intercellular spaces, retains cells based on the formation of a jelly-like matrix in tissues, retains the lubricity and flexibility of organ tissues, and causes mechanical disorders. It has many functions such as resistance to external force and prevention of bacterial infection (BIO IND
USTRY, Vol. 8, 346, 1991).

Furthermore, it has recently been found that hyaluronic acid has various physiological actions depending on its molecular weight.
For example, the high molecular weight hyaluronic acid (molecular weight of 1,000,000 or more), which is considered to be synthesized in vivo, has proteoglycan release inhibitory action, bradykinin joint pain inhibitory action, damaged cartilage repairing action, and arthritis inhibitory action, thus suppressing inflammation. Effective (BIO INDUSTRY, Volume 11, 6
32, 1991), its degradation (low molecular weight) product, low molecular weight hyaluronic acid, promotes angiogenesis (Scie
No. 228, page 1324, 1985), and since it has a leukocyte chemotaxis promoting action (Japanese Patent Publication No. 6-8323), it is considered to have an inflammation promoting action.

From the above, low molecular weight hyaluronic acid is associated with hepatitis, gingivitis (inflammation, vol. 4, p. 437, 1984),
Rheumatoid arthritis, osteoarthritis (connective tissue, 25, 24
Page 3, 1994), Malignant tumor (J. Cellular Physiolo)
gy, 160, 275, 1994), which is considered to be closely related to the exacerbation of symptoms, and therefore, inhibition of hyaluronic acid degradation may be effective in the treatment of these diseases.

[0006] Actually, hyaluronidase, which lowers the molecular weight of hyaluronic acid in connective tissue, is supposed as an enzyme having an inflammation-promoting action, and therefore, an inhibitor of the hyaluronidase was expected to have a pharmacological effect (inflammation, vol. 4, vol. 437
P., 1984).

By the way, there have been many reports on hyaluronidase inhibitors from the past (Japanese Patent Publication No.
No. 29271, Japanese Patent Publication No. 6-4584, Japanese Unexamined Patent Publication No. 5-178876, Japanese Unexamined Patent Publication No. 6-80553, Japanese Unexamined Patent Publication No. 6-80576, and Japanese Unexamined Patent Publication No. 6-9415.
JP-A-6-9416, JP-A-3-685
All of these inhibitors are inhibitors of bovine testis-derived hyaluronidase. This is because there is no report that hyaluronidase was isolated from fibroblasts forming human connective tissue, and all of the above inhibitors have the same properties as those of bovine testis-derived hyaluronidase derived from human fibroblasts. It is assumed that they have it, and they have only searched for various inhibitors against this bovine testis-derived hyaluronidase.

However, in recent years, human synovial cells present in joints (connective tissue, 25, 243, 1994), human uterine cervix cells (FEBS Letters, 347, 95, 1994)
), Human skin fibroblasts (BBA, 172, 70, 1
(990), it is clearly different from the degradation mechanism by the lysosomal hyaluronidase from bovine testis, which is a weakly acidic, endo-type and decomposes hyaluronic acid into tetra- and hexasaccharides.
It has been reported that hyaluronic acid decomposes extracellularly into about 200 sugars.

[0009]

The inventors of the present invention have conducted extensive studies and found that testicular hyaluronidase cannot inhibit the hyaluronic acid degradation of connective tissue-derived hyaluronidase and completed the present invention. However, the purpose thereof is to act on cells present in human connective tissue, and to inhibit hyaluronic acid degradation, a hyaluronic acid degradation inhibitor, and further excellent therapeutic effect for diseases associated with the degradation of hyaluronic acid. It is intended to provide a therapeutic agent for a hyaluronic acid-degrading disease.

[0010]

The above-described object is to provide a hyaluronic acid decomposition inhibitor characterized by containing a histamine H ₁ -antagonist or a salt thereof, or a therapeutic agent for hyaluronic acid decomposition disease containing the inhibitor. To be achieved.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION The structure of the present invention will be described in detail below.

Examples of the histamine H ₁ -antagonist used in the present invention include ethanolamine-type diphenhydramine, ethylenediamine-type pyrilamine, triprenamine, alkylamine-type chlorpheniramine,
Brompheniramine, piperazine-type cyclidine, phenothiazine-type promethazine and the like can be mentioned.

The salt of histamine H ₁ -antagonist is not particularly limited, but particularly, pharmaceutically acceptable inorganic acid salts such as hydrochloride, hydrobromide, sulfate and phosphate, and acetate. , Fumarate, maleate, tartrate, citrate, p-toluenesulfonate, and other organic acid salts.

These histamine H ₁ -antagonists or salts thereof can be used alone or in admixture of two or more.

The content of the histamine H ₁ -antagonist or the salt thereof in the hyaluronic acid degradation inhibitor or the therapeutic agent for hyaluronic acid degradation disease of the present invention is determined by the type and degree of the target disease, the age, weight, sex of the patient, etc. However, it is generally 0.0001 to 50 based on the total amount of the applied composition as 100 g.
g is preferable, and 0.001 to 5 g is particularly preferable.

When the hyaluronic acid decomposition inhibitor of the present invention is used in combination with, for example, a therapeutic agent for arthritis, which will be described later, when it is combined with a high molecular weight hyaluronic acid, the therapeutic effect is further improved. The high molecular weight hyaluronic acid used is preferably deproteinized and highly pure, and preferably has a molecular weight of 5
It is 100,000 or more, more preferably 800,000 or more, and does not depend on the object of extraction or the manufacturing method. Further, the blending amount thereof is preferably 0.1 to 10% by weight, and particularly preferably 0.5 to 2% by weight, based on the preparation. Further, the obtained high molecular hyaluronic acid may be modified by a crosslinking treatment or the like.

The hyaluronic acid decomposition inhibitor and the therapeutic agent for hyaluronic acid decomposition disease of the present invention may be used in the form of tablets, liquids, capsules, using appropriate excipients, carriers and diluents.
Granules, powders, ointments, patches, injections, joint injections,
Examples thereof include suppositories, bath agents and the like, and gels, creams, sprays, patches, lotions, packs, emulsions, powders and bath agents.

The preparation of such a preparation is carried out by a conventional method,
For example, solid preparations are formulated using usual pharmaceutical additives such as lactose, starch, crystalline cellulose, talc and the like. Capsules can be obtained by filling the thus-prepared fine granules, powders and the like into suitable capsules. The liquid preparation is prepared by dissolving and suspending the agent of the present invention in an aqueous solution containing sucrose, carboxymethylcellulose and the like.

The excipient or auxiliary agent to be added to the hyaluronic acid decomposition inhibitor or the therapeutic agent for hyaluronic acid decomposition disease of the present invention may be one normally used in cosmetics, inhibitors, etc. It is appropriately selected according to, but is not particularly limited, for example, hydrocarbons such as petrolatum and squalane, higher alcohols such as stearyl alcohol, higher fatty acid lower alkyl esters such as isopropyl myristate, animal fats and oils such as lanolinic acid. , Polyhydric alcohols such as glycerin and propylene glycol, glycerin fatty acid esters, polyethylene glycol monostearate, surfactants such as polyethylene alkyl ether phosphate, preservatives such as methyl paraoxybenzoate and butyl paraoxybenzoate, waxes, resins, Various flavors, various pigments, sodium citrate Beam, sodium carbonate, various inorganic salts and various acids such as lactic acid, like water, and ethanol and the like.

The hyaluronic acid decomposition inhibitor of the present invention can be incorporated as an active ingredient of a therapeutic agent for a disease in which the decomposition of hyaluronic acid is accelerated. Further, it can be used as an active ingredient of ordinary medicines and cosmetics, and also as a research / test reagent added to a cultured cell system.

In the present invention, the therapeutic agent includes a so-called therapeutic agent that removes the symptoms of the diseases described below, an improving agent that reduces the symptoms, and a preventive agent that prevents the appearance of the symptoms.

The term "disease" as used in the present invention refers to a condition in which the hyaluronic acid decomposition is physiologically higher than normal or the hyaluronic acid is abnormally decomposed.

Examples of diseases in which the hyaluronic acid decomposition is physiologically higher than normal are gingivitis, dry skin and rough skin. When the therapeutic agent for hyaluronic acid-degrading diseases of the present invention is used for these diseases, it can be applied as a preventive agent to normal persons, but it is applied to those who have symptoms in which the decomposition of hyaluronic acid is physiologically higher than normal. It is more effective to do so.

Diseases in which hyaluronic acid is abnormally decomposed include rheumatism, in which the decomposition of hyaluronic acid is abnormally increased in the affected area, osteoarthritis, psoriatic arthropathy, gout, polyarthritis, traumatic arthritis, etc. Arthrosis, hepatitis, gingivitis, and malignant tumors, and cirrhosis, transplant rejection, psoriasis, and psoriasis, which are considered to increase abnormally hyaluronan in the affected area due to increased hyaluronan levels in serum. Scleroderma, fibrosis such as liver cirrhosis where the organs are hardened due to diseases, arteriosclerosis, etc., and xeroderma, dry skin, rough skin, and other nephritis in which the water retention capacity is reduced in the affected area as a result of hyaluronic acid decomposition. , Keloid, hyperrepair, sepsis and other diseases. When the therapeutic agent for a hyaluronic acid-degrading disease of the present invention is used for these diseases, it can be applied as a symptom-improving agent or therapeutic agent to a person with symptom in which the decomposition of hyaluronic acid is abnormally enhanced.

Among these diseases in which hyaluronic acid is abnormally decomposed, it is more effective especially in diseases in which the amount of histamine in the serum of patients is increased. Specifically, rheumatism, osteoarthritis, Psoriasis, such as psoriatic arthrosis, gout, polyarthritis, and traumatic arthritis
Examples include scleroderma, keloids, malignant tumors, and transplant rejection.

At this time, histamine acts not only on vascular endothelial cells and smooth muscle cells but also on fibroblasts and synovial cells present in connective tissues, and decomposes hyaluronic acid which is a component of the intercellular matrix. It was revealed that the present invention has an action, and it was revealed by the present invention that the H ₁ -antagonist suppresses this action.

In an allergic disease in which histamine is considered to play an important role, it is considered that hyaluronic acid has a low molecular weight at the lesion site and promotes inflammation.
It is considered that the histamine H ₁ -antagonist currently used in the allergic disease has a connective tissue stabilizing effect in addition to the known effect.

Therefore, the therapeutic agent for hyaluronic acid-degrading diseases of the present invention can be expected to treat not only allergic diseases but also diseases associated with abnormal decomposition of hyaluronic acid in connective tissues and joints.

As a method for administering the therapeutic agent for hyaluronic acid-degrading disease of the present invention, oral administration such as tablets and capsules or parenteral administration such as injection is possible.

Particularly when used for the improvement or treatment of arthropathy, parenteral administration by injection or injection injecting directly into the joint is preferable because the therapeutic effect can be expected.

When used for the treatment of arthropathy or the like, it may be premixed with high molecular weight hyaluronic acid, for example, a commercially available hyaluronic acid preparation (Alz, Kaken Pharmaceutical Co., etc.) or separately injected at the same time as administration. Therefore, it is effective to use them simultaneously, and further therapeutic effects are expected by injecting them directly into the joint together with high molecular weight hyaluronic acid.

One of the therapeutic agents for hyaluronan-degrading diseases of the present invention
The daily dose is usually 10 μm as histamine H ₁ -antagonist or its salt by oral administration.
g to 2 g is preferable, and 50 μg to 100 is particularly preferable.
mg. For parenteral administration, it is preferably 500 μg to 1 g.

The hyaluronic acid degradation inhibitor of the present invention, when added to cultured cells to produce high molecular weight hyaluronic acid,
It is preferably added so as to be 1 nM or more in the culture medium, more preferably 0.01 μM to 1 mM.

The therapeutic agent for hyaluronic acid-degrading diseases of the present invention is
It is preferable to use after sterilizing by a method such as heat sterilization or filtration sterilization.

[0035]

EXAMPLES The present invention will be described in more detail with reference to Examples and Comparative Examples. An evaluation system for investigating the effect of the hyaluronic acid decomposition inhibitor will be described prior to Examples. Moreover, the compounding amounts shown in Tables 3 to 13 all represent% by weight.

(1) Method for preparing MEM medium Minimum Essential Medium
(Dainippon Pharmaceutical Co., Ltd., 10-101) 10.6 g each with a final concentration of 1% (V / V) Non Essentia
l Amino Acid (Dainippon Pharmaceutical Co., 16-81
0) 1 mM sodium pyruvate (manufactured by Dainippon Pharmaceutical Co., Ltd., 1
6-820), 1.2% (W / V) sodium hydrogencarbonate and distilled water were added to make 1 liter, and then carbon dioxide gas was blown to bring the pH to about 7 (hereinafter abbreviated as MEM medium).

(2) Inactivation of fetal bovine serum (FBS) FBS (manufactured by Irvine Scientific)
Heat treatment was performed at 6 ° C. for 30 minutes.

(3) Method for Preparing Polymer Tritium Hyaluronic Acid for Cell Addition Normal human fibroblast cell line [Detroit 551 strain (ATCC
CCL 110)] was adjusted to 2 × 10 ⁵ cells / ml with MEM medium containing 10% (V / V) inactivated FBS, 50 ml was put in a 225 cm ² flask and cultured for 3 days to be confluent. I chose Then, the precursor of hyaluronic acid, tritium glucosamine (American Rad
iolabeled Chemicals Inc.) was added to the culture system (10 μm).
Ci / ml), after further culturing for 3 days, the tritiated hyaluronic acid from the culture solution was purified by the method of Underhill et al. (J. Cell Biology, Vol. 82, 475, 1979) and further by gel filtration column. Molecular weight 10
High molecular weight tritium hyaluronic acid (specific radioactivity, 0.1 μCi / μg) of more than 0,000 was prepared. This was used as polymer tritium hyaluronic acid for addition to the cell culture system.

(4) Culture of normal human fibroblasts with addition of polymeric tritium hyaluronan Normal human fibroblast cell line [Detroit 551 strain (ATCC
CCL 110)], the cell number is 1.5 × 10 ⁵ cells / m 2 in MEM medium containing 10% (V / V) inactivated FBS.
Adjust to 1 and 0.8 on a 12-well plate (Falcon)
ml seeds, 95% (V / V) air-5% (V / V
V) Incubation at 37 ° C. for 3 days in an atmosphere of carbon dioxide gas,
Further, the medium was replaced with only the MEM medium, and the cells were cultured for 1 day. Then, the polymer containing tritium hyaluronan (1
4,000 DPM / ml) MEM medium was prepared, the medium was exchanged, and the cells were cultured for 3 days. When the medium was replaced, histamine and various antagonists were added so that the final concentration was 10 μM.

(5) Addition culture of high molecular weight tritium hyaluronan to rheumatoid synovial cells 1.5 × 10 ^{5 of} human rheumatoid synovial cells in MEM medium containing 10% (V / V) inactivated FBS. Pieces / m
Adjust to 1 and 0.8 on a 12-well plate (Falcon)
ml seeds, 95% (V / V) air-5% (V / V
V) Incubation at 37 ° C. for 3 days in an atmosphere of carbon dioxide gas,
Further, the medium was replaced only with the MEM medium containing 1% FBS, and the cells were cultured for 1 day. Then, a MEM medium containing high-molecular tritium hyaluronic acid (14,000 DPM / ml) was prepared, the medium was exchanged, and culturing was carried out for 3 days. When the medium was exchanged, histamine, various antagonists, and cold high molecular weight hyaluronic acid were added so that the final concentration was 10 μM.

(6) Evaluation of Degradation of Polymer Tritium Hyaluronic Acid by Cells After the completion of the culture, the culture solution was collected and heat-treated at 100 ° C. for 5 minutes, and 1 ml of the medium was added to a Sepharose CL-2B column (inner diameter: 1 cm, (Length 60 cm) was applied and gel filtration was performed under the following conditions. Flow rate: 0.6 ml / min Fraction: 4 ml / 1 Fraction Total number: 25 Further, 26 fractions 10 to 25 in which hyaluronic acid having a molecular weight of 100,000 or less was eluted were collected, and [ ³ H] radioactivity was measured and decomposed. The amount of hyaluronic acid was determined. Furthermore, the hyaluronic acid decomposition rate and the decomposition inhibition rate were determined by the following equations 1 and 2.

[0042]

[Equation 1] Degradation rate of hyaluronic acid (%) = B / A x 100 A = Degradation amount of hyaluronic acid promoted by addition of histamine B = Degradation amount of hyaluronic acid promoted by simultaneous addition of histamine + drug

[0043]

[Equation 2] Degradation inhibition rate (%) = (1−B / A) × 100 A = Amount of hyaluronic acid decomposition promoted by addition of histamine B = Amount of hyaluronic acid decomposition promoted by simultaneous addition of histamine + drug

Examples 1 to 3 and Comparative Examples 1 to 2 Chlorpheniramine, pyrilamine and diphenhydramine (3.9 mg, 4.0 mg and 2.9 mg) were dissolved in 10 g of purified water to prepare respective aqueous solutions ( Examples 1-3). Moreover, purified water was used as Comparative Example 1, and glycyrrhizin (inflammation, 4 volumes, NO4, 437 (1), which is a conventionally known inhibitor of bovine testis-derived hyaluronidase, was used.
984)) 0.18 g was dissolved in 10 g of water to prepare an aqueous solution, which was used as Comparative Example 2.

Test Example 1 (Hyaluronic Acid Degradation by Human Normal Skin Fibroblasts) Polymer tritium was prepared by the method (6) described above using Examples 1 to 3, Comparative Example 1 (aqueous solution) and Comparative Example 2. The decomposition of hyaluronic acid was examined, and the decomposition rate of hyaluronic acid and the inhibition rate of decomposition of hyaluronic acid were calculated from the above formulas 1 and 2. The results are shown in Table 1.

[0046]

[Table 1]

As a result, the degradation of hyaluronic acid was promoted by the addition of histamine more than when it was not added, and inhibition of the degradation was observed in any of the drugs chlorpheniramine, pyrilamine and diphenhydramine (Examples 1 to 3) which are antagonists. It was It was also found that the addition of glycyrrhizin, which is an inhibitor of bovine testis-derived hyaluronidase shown in Comparative Example 1 or Comparative Example 2, had no effect in the present evaluation system which is a human cell culture system.

From these results, it was found that the histamine H ₁ -antagonist is effective as a human hyaluronic acid degradation inhibitor. Further, it is considered that the hyaluronic acid decomposition inhibitor of the present invention can be used as an active ingredient of a therapeutic agent for a disease in which hyaluronic acid decomposition is abnormally enhanced.

Examples 4 to 8 and Comparative Examples 3 to 6 H ₁ -antagonist chlorpheniramine was dissolved in water so that the concentrations in the medium were 0.1, 1.0 and 10 μM, respectively. An aqueous solution was prepared (Example 4 to
6). Similarly, pyrilamine and diphenhydramine were dissolved in water so that the concentration in the medium was 10 μM to prepare respective aqueous solutions (Examples 7 to 8). In addition, purified water was used as Comparative Example 3 and Comparative Examples 4 and 5, respectively, H ₂ -antagonist cimetidine and H ₃ -antagonist thioperamide were dissolved in water to 10 μM in the medium, and the aqueous solution was filtered. Sterilized and conditioned. Further, as Comparative Example 6, high molecular weight hyaluronic acid (derived from human umbilical cord, molecular weight 800,000 to 1,200,000, Seikagaku Corporation) was added to the medium at a final concentration of 1 m.
It was dissolved in water so as to be g / ml, and the aqueous solution was sterilized by filtration to prepare.

Test Example 2 (Hyaluronic Acid Degradation by Human Rheumatoid Synovial Cells) Using Examples 4 to 8 and Comparative Example 3 (purified water), 4 to 6,
The decomposition of the polymeric tritiated hyaluronic acid was examined by the method (6) described above, and the hyaluronic acid decomposition rate and the hyaluronic acid decomposition inhibition rate were calculated from the above equations 1 and 2. Table 2 shows the results.

[0051]

[Table 2]

As a result, the decomposition of hyaluronic acid promoted by the addition of histamine was not completely inhibited by the H ₁ -antagonists chlorpheniramine, pyrilamine and diphenhydramine (Examples 4 to 8). Admitted. However, it was not inhibited at all by the H ₂ -antagonist cimetidine and the H ₃ -antagonist thioperamide (Comparative Examples 4 and 5). From this fact, the promotion of the degradation of hyaluronic acid by histamine is H _1-.
It is a receptor-mediated reaction, and it is clear that its degradation is suppressed only by H ₁ -antagonist among histamine antagonists. Furthermore, the addition of high molecular weight hyaluronic acid, which is the main component of a hyaluronic acid preparation known as a therapeutic agent for arthropathy, to the culture system slightly suppressed the promotion of the degradation of hyaluronic acid by histamine.
_It was significantly smaller than the effect of ₁ -antagonist.

From these results, the histamine H ₁ -antagonist is effective as a human hyaluronic acid degradation inhibitor.
Further, the therapeutic agent for hyaluronic acid-degrading disease containing the hyaluronic acid-degrading inhibitor of the present invention is considered to be effective for treating arthrosis and the like in which hyaluronic acid decomposition is abnormally enhanced.

Examples 9 to 11 (tablets)

[0055]

[Table 3]

The above components are uniformly mixed and compressed into tablets to give 170 mg per tablet according to a conventional method.

Examples 12 to 14 (capsules)

[0058]

[Table 4]

The above components are uniformly mixed, and No. 3 hard capsules are filled with 150 mg of the mixture according to a conventional method.

Examples 15 to 17 (solution)

[0061]

[Table 5]

The above components are dissolved in purified water, and the mixture is stirred and homogenized to obtain a liquid agent.

Examples 18 to 21 (cream)

[0064]

[Table 6]

The component (A) was uniformly mixed and dissolved at 80 ° C., and then the component (B) was mixed and dissolved (mixed solution I). Separately from this, the component (D) was uniformly mixed and dissolved at 80 ° C., and then the component (C) was mixed and dissolved therein (mixed liquid II). Next, the mixed solution II was gradually added to the mixed solution I and cooled to 30 ° C. with sufficient stirring to obtain a cream.

Examples 22 to 25 (lotion)

[0067]

[Table 7]

A lotion was prepared by mixing and dissolving each component.

Examples 26 to 27 (Bathing agent)

[0070]

[Table 8]

Bath ingredients were prepared by mixing the components. In addition, this bathing agent is diluted about 3000 times at the time of use.

Examples 28-31 (ointment)

[0073]

[Table 9]

The components of (B) above were mixed while heating to 80 ° C. in a water bath, and the mixture was heated to 80 ° C. above (A).
Was slowly added to the mixture of the respective components under stirring. Next, a homogenizer (Tokusyukika Kog)
vigorously stirred for 2.5 minutes (2500 rpm)
Then, each component was sufficiently emulsified and dispersed, and then gradually cooled with stirring to obtain an ointment containing promethazine or diphenhydramine.

Examples 32-34 (toothpaste)

[0076]

[Table 10]

Purified water, glycerin, carrageenan, saccharin, butyl paraoxybenzoate, chlorhexidine diglyconate, a fragrance and an antagonist were weighed and mixed according to a conventional method to swell the binder, and then dibasic calcium phosphate and lauryl sulfate. Sodium was added, and the mixture was further thoroughly mixed and defoamed, and then filled in a tube to obtain a toothpaste.

Examples 35 to 37 (mouthwash)

[0079]

[Table 11]

A mouthwash was obtained according to a conventional method.

Examples 38-40 (injection)

[0082]

[Table 12]

Diphenhydramine, chlorpheniramine, sodium chloride and sodium monohydrogen phosphate were placed in a graduated cylinder, distilled water for injection was added, dissolved, filtered through a membrane filter (0.22 μm), and dispensed into a glass ampoule.

Examples 41 to 43 (joint injection)

[0085]

[Table 13]

An aqueous solution of high molecular weight hyaluronic acid having a concentration twice that shown in Table 13 was sterilized by heating (solution A) to prepare an aqueous solution (solution B) in which diphenhydramine, pyrilamine and sodium chloride having a concentration twice were dissolved. . Next, the liquid A and the liquid B were mixed at a ratio of 1: 1 under aseptic conditions, stirred well, and then dispensed into injection syringes at 2.5 ml each.

[0087]

INDUSTRIAL APPLICABILITY As described above, it is apparent that the present invention can provide a hyaluronic acid degradation inhibitor that acts on cells present in human connective tissue and inhibits hyaluronic acid degradation, and a therapeutic agent for hyaluronic acid degradation disease. is there.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 7/48 A61K 7/48 7/50 7/50 31/44 31/44 ABX ABX ACS ACS ADA ADA 31/495 ACK 31/495 ACK 31/54 31/54 (72) Inventor Shintaro Inoue 5-3 28, Shoucho, Odawara-shi, Kanagawa Kanebo Co., Ltd.

Claims (3)

[Claims] 1. A hyaluronic acid degradation inhibitor, which comprises a histamine H ₁ -antagonist or a salt thereof. 2. The hyaluronic acid degradation inhibitor according to claim 1, wherein the histamine H ₁ -antagonist is chlorpheniramine, pyrilamine, diphenhydramine, cyclidine, promethazine, brompheniramine, or triprenamine. 3. A therapeutic agent for a hyaluronic acid-degrading disease, which comprises the hyaluronic acid-degrading inhibitor according to claim 1 or 2.