JP7799479B2 - Composition containing alkyldiaminoethylglycine and/or its salt - Google Patents

Description

The present disclosure relates to compositions containing alkyldiaminoethylglycine and/or salts thereof, as well as uses thereof.

Matrix metalloproteinase (MMP) is a general term for extracellular matrix degrading enzymes that contain a zinc(II) ion at their active site. Their main substrates are biopolymers such as collagen, laminin, gelatin, and fibronectin.

MMPs are thought to maintain homeostasis by breaking down unnecessary extracellular matrix and controlling angiogenesis and the construction of new tissue during the processes of tissue remodeling and wound healing in the body. However, excessive MMP activity can lead to the progression of tissue destruction, and MMPs are thought to be involved in periodontal disease, rheumatoid arthritis, tumor invasion and metastasis, among other phenomena. Therefore, compounds that can control MMP activity are being sought.

Patent No. 5339708

Int J Environ Res Public Health. 2020 Jul;17(14):4923. Folia Pharmacol. Jpn. 141, 314-320 (2013) J Periodontal Res. 1994 Jul;29(4):290-8. J Clin Periodontol. 2021 Aug; 48(8):1051-1065.

MMP-8, also known as neutrophil collagenase because it is a collagenase produced by neutrophils, is known to primarily degrade extracellular matrix such as collagen and accounts for approximately 80% of the collagenase in gingival crevicular fluid. Furthermore, approximately 60% of periodontal tissues are composed of collagen fibers, which provide the connective tissue attachment that supports the teeth. Neutrophils accumulate in gingival crevicular fluid in response to inflammation in the periodontal pocket and produce neutrophil collagenase (MMP-8) (Non-Patent Document 1). This destroys collagen fibers in the periodontal tissue, resulting in the loss of connective tissue attachment and attachment loss (Non-Patent Document 2). Furthermore, attachment loss is thought to lead to deepening of periodontal pockets and gingival recession (Non-Patent Document 3). Therefore, there is a need to identify components that can control the activity of neutrophil collagenase (Patent Document 1).

The objective of the present disclosure is to provide a composition for inhibiting neutrophil collagenase activity.

The inventors discovered that alkyldiaminoethylglycine hydrochloride has inhibitory activity against neutrophil collagenase and have made further improvements.

The present disclosure encompasses, for example, the subject matter described in the following sections:
Item 1.
A composition for inhibiting periodontal pocket formation and/or periodontal pocket deepening, which comprises alkyldiaminoethylglycine and/or a salt thereof as an active ingredient.
Item 2.
A composition for inhibiting gingival collagen degradation, comprising alkyldiaminoethylglycine and/or a salt thereof as an active ingredient.
Item 3.
A composition for inhibiting neutrophil collagenase activity, comprising alkyldiaminoethylglycine and/or a salt thereof as an active ingredient.
Item 4.
Item 4. The composition according to any one of Items 1 to 3, which is used for at least one purpose selected from the group consisting of inhibiting gingival recession, inhibiting periodontal tissue destruction, inhibiting attachment loss, and inhibiting gum recession.
Item 5.
Item 5. The composition according to any one of Items 1 to 4, wherein the total content of alkyldiaminoethylglycine and/or a salt thereof in the composition is 0.002% by mass or more.
Item 6.
Item 6. The composition according to any one of Items 1 to 5, which is an oral composition.

A composition for inhibiting neutrophil collagenase activity is provided. Also provided is a composition for inhibiting periodontal pocket formation. Also provided is a composition for inhibiting periodontal pocket deepening. Also provided is a composition for inhibiting gingival collagen degradation.

The results of measuring neutrophil collagenase (MMP-8) activity when each material was added are shown. 1 shows the results of measuring neutrophil collagenase activity when alkyldiaminoethylglycine hydrochloride was added.

Each embodiment included in the present disclosure will be described in further detail below.
The composition encompassed by the present disclosure contains alkyldiaminoethylglycine and/or a salt thereof. In this specification, the composition may be referred to as the "composition of the present disclosure."

Alkyldiaminoethylglycine has the following formula:

In the formula, R is C ₈ H ₁₇ to C ₁₆ H ₃₃ , mainly C ₁₂ H ₂₅ and C ₁₄ H _29. The total amount of C ₁₂ H ₂₅ and C ₁₄ H ₂₉ is preferably 50, 60, 70, 80, or 90% or more of the alkyldiaminoethylglycine.

Salts of alkyldiaminoethylglycine include, for example, inorganic acid salts, organic acid salts, and metal salts. Examples of inorganic acid salts include hydrochloride and sulfate. Examples of organic acid salts include tartrate and acetate. Examples of metal salts include sodium salt, potassium salt, calcium salt, and magnesium salt. Among these, alkyldiaminoethylglycine hydrochloride is preferred.

The total content of alkyldiaminoethylglycine and/or a salt thereof in the composition of the present disclosure is not particularly limited as long as the effect is achieved. For example, the total content of alkyldiaminoethylglycine and/or a salt thereof in the composition of the present disclosure can be 0.0005% by mass or more. The upper or lower limit of this range may be, for example, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, or 0.2% by mass. For example, the content may be 0.001 to 0.1% by mass or 0.005 to 0.05% by mass.
Although not particularly limited, from the viewpoint of achieving even stronger inhibitory activity of alkyldiaminoethylglycine and/or its salts against neutrophil collagenase activity, the total content of alkyldiaminoethylglycine and/or its salts is preferably 0.0005% by mass or more, and more preferably 0.005% by mass or more.

Neutrophil collagenase (EC 3.4.24.34) is a collagenase produced by neutrophils and is also known as MMP-8.

Inhibition of neutrophil collagenase activity means inhibiting the decomposition of substrates (e.g., collagens such as type I collagen, type II collagen, and type III collagen) by neutrophil collagenase.

Since alkyldiaminoethylglycine and/or its salts inhibit neutrophil collagenase activity, the composition of the present disclosure is suitable as a composition for inhibiting neutrophil collagenase activity. Furthermore, alkyldiaminoethylglycine and/or its salts inhibit the degradation of substrates by neutrophil collagenase, thereby suppressing the destruction of tissues (e.g., junctional epithelium, gingival sulcus epithelium, periodontal ligament, etc.) and the degradation of gingival collagen (collagen fibers that make up the gingiva) by neutrophil collagenase. Furthermore, since the composition of the present disclosure has the effect of inhibiting neutrophil collagenase activity, it can inhibit the activity of neutrophil collagenase that has already been produced.

For this reason, the composition of the present disclosure is suitable as an oral composition. More specifically, the composition of the present disclosure can be used for, for example, inhibiting gingival recession, inhibiting periodontal pocket formation, inhibiting periodontal pocket deepening, inhibiting gingival destruction, inhibiting periodontal tissue destruction, inhibiting gingival recession, inhibiting gingival collagen degradation, inhibiting attachment loss, protecting periodontal tissue collagen, inhibiting periodontal tissue destruction associated with chronic inflammation, and/or protecting dentin collagen.
As used herein, "inhibition of periodontal pocket formation" means inhibiting the formation of periodontal pockets (grooves between the teeth and gums). As used herein, "inhibition of periodontal pocket deepening" means inhibiting the deepening of periodontal pockets (grooves between the teeth and gums). As used herein, "inhibition of attachment loss" means inhibiting the gingival attachment epithelium from peeling off from the tooth surface, causing the attachment position between the gums and the teeth to shift toward the tooth root. As used herein, "inhibition of gingival recession" means inhibiting the entire gums from moving toward the tooth root. As used herein, "inhibition of gum recession" means inhibiting the exposure of the tooth root due to the movement of the entire gums toward the tooth root.

The composition of the present disclosure is preferably applied to subjects with elevated neutrophil collagenase activity. More specifically, examples include individuals who fall into stage 1, 2, 3, or 4, or grade C, of the diagnostic criteria for periodontal disease (Non-Patent Document 4). The diagnostic criteria for periodontal disease were developed at a workshop co-sponsored by the American Academy of Periodontology and the European Federation of Periodontology, and can be found on the Japanese Society of Periodontology website (https://www.perio.jp/file/news/info_191220.pdf). Examples of individuals who fall into stage 1 of the diagnostic criteria for periodontal disease include those with a maximum interdental clinical attachment level (the distance from the cement-enamel junction to the gingival sulcus epithelium (measured in 1 mm increments)) of approximately 1 to 2 mm. Examples of individuals who fall into stage 2 of the diagnostic criteria for periodontal disease include those with a maximum interdental clinical attachment level of approximately 3 to 4 mm. Examples of people who fall into stage 3 or 4 according to the diagnostic criteria for periodontal disease include people whose largest interdental clinical attachment level is 5 mm or more. Among these, examples of people who fall into stage 3 according to the diagnostic criteria for periodontal disease include people who have lost 1 to 4 teeth due to periodontal disease. Examples of people who fall into stage 4 according to the diagnostic criteria for periodontal disease include people who have lost 5 or more teeth due to periodontal disease. Examples of people who fall into grade C according to the diagnostic criteria for periodontal disease include people whose clinical attachment level has changed by 2 mm or more over 5 years.
Furthermore, examples of subjects to which the composition of the present disclosure can be applied include mammals including humans (for example, dogs, cats, mice, rats, sheep, horses, cows, monkeys, etc.), with humans being preferred.

The composition of the present disclosure may be, for example, a solid composition, a liquid composition, etc. Furthermore, the composition of the present disclosure (particularly an oral composition) can be made into the form (dosage form) of, for example, an ointment, paste, paste, gel, liquid, spray, mouthwash, liquid dentifrice, toothpaste, gum, tablet, drop, etc. in accordance with conventional methods. Of these, mouthwash, liquid dentifrice, toothpaste, ointment, paste, liquid, or gel form is preferred.

The composition of the present disclosure may further contain, for example, one or more optional components that can be incorporated into oral compositions, as long as the effects are not impaired.

For example, nonionic surfactants, anionic surfactants, or amphoteric surfactants can be used as surfactants. Specific examples of nonionic surfactants include sugar fatty acid esters such as sucrose fatty acid esters, maltose fatty acid esters, and lactose fatty acid esters; fatty acid alkanolamides; glycerin fatty acid esters; sorbitan fatty acid esters; fatty acid monoglycerides; polyoxyethylene alkyl ethers having a polyoxyethylene addition coefficient of 8 to 10 and an alkyl group containing 13 to 15 carbon atoms; polyoxyethylene alkylphenyl ethers having a polyoxyethylene addition coefficient of 10 to 18 and an alkyl group containing 9 carbon atoms; diethyl sebacate; polyoxyethylene hydrogenated castor oil; and fatty acid polyoxyethylene sorbitan. Examples of anionic surfactants include sulfate ester salts such as sodium lauryl sulfate and polyoxyethylene lauryl ether sodium sulfate; sulfosuccinates such as sodium lauryl sulfosuccinate and polyoxyethylene lauryl ether sodium sulfosuccinate; acylamino acid salts such as sodium cocoyl sarcosinate and sodium lauroyl methyl alanine; and sodium cocoyl methyl taurate. Examples of zwitterionic surfactants include acetate betaine surfactants such as lauryl dimethylaminoacetate betaine and coconut oil fatty acid amidopropyl dimethylaminoacetate betaine; and imidazoline surfactants such as N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine sodium. These surfactants can be used alone or in combination of two or more. The amount used is typically 0.1 to 5% by mass of the total composition.

Flavors that can be used include, for example, menthol, carvone, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronellol, α-terpineol, methyl acetate, citronenyl acetate, methyl eugenol, cineole, linalool, ethyl linalool, thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, cinnamon oil, perilla oil, wintergreen oil, clove oil, eucalyptus oil, pimento oil, d-camphor, d-borneol, fennel oil, cinnamon oil, cinnamonaldehyde, peppermint oil, and vanillin. These may be blended alone or in combination of two or more types, at a concentration of, for example, 0.001 to 1.5% by mass based on the total amount of the composition.

Sweetening agents that can be used include, for example, saccharin sodium, acesulfame potassium, stevioside, neohesperidyl dihydrochalcone, perillartine, thaumatin, aspartyl phenylalanyl methyl ester, and p-methoxycinnamic aldehyde. These can be incorporated in amounts of, for example, 0.01 to 1% by mass of the total composition.

In addition, humectants such as sorbitol, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactite, and polyoxyethylene glycol can be blended alone or in combination of two or more.

Examples of binders include cellulose derivatives such as sodium carboxymethylcellulose, carboxymethylethylcellulose salts, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, ethylcellulose, crystalline cellulose, and crystalline cellulose-carmellose sodium; microbially produced polymers such as xanthan gum; natural polymers or natural rubbers such as tragacanth gum, karaya gum, gum arabic, carrageenan, dextrin, agar, pectin, pullulan, gellan gum, locust bean gum, and sodium alginate; synthetic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, polyvinyl methyl ether, and sodium polyacrylate; thickening silica; inorganic binders such as Veegum; and cationic binders such as O-[2-hydroxy-3-(trimethylammonio)propyl]hydroxyethylcellulose chloride. These binders can be used alone or in combination of two or more.

Preservatives that can be added include parabens such as methylparaben, ethylparaben, propylparaben, and butylparaben, as well as sodium benzoate and phenoxyethanol.

Coloring agents may include legal pigments such as Blue No. 1, Yellow No. 4, Red No. 202, and Green No. 3, mineral pigments such as ultramarine, enhanced ultramarine, and Prussian blue, and titanium oxide.

pH adjusters such as citric acid, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, glycerophosphoric acid, acetic acid, nitric acid, or chemically acceptable salts thereof, or sodium hydroxide may be added. These may be added alone or in combination of two or more types so that the pH of the composition is in the range of 4 to 8, preferably 5 to 7. The amount of pH adjuster added may be, for example, 0.01 to 2% by weight.

The oral composition of the present disclosure may further contain, as active ingredients, vitamin E compounds such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate; amphoteric bactericides such as dodecyldiaminoethylglycine; nonionic bactericides such as triclosan and isopropylmethylphenol; anionic bactericides such as sodium lauroyl sarcosinate; cationic bactericides such as cetylpyridinium chloride, chlorhexidine hydrochloride, benzalkonium chloride, and benzethonium chloride; dextranase, amylase, protease, mutanase, lysozyme, and lytic enzymes (retic enzymes). Enzymes, alkali metal monofluorophosphates such as sodium monofluorophosphate and potassium monofluorophosphate, fluorides such as sodium fluoride and stannous fluoride, tranexamic acid, epsilon aminocaproic acid, aluminum chlorohydroxyl allantoin, dihydrocholesterol, hinokitiol, glycyrrhetinic acid, glycyrrhizinic acid, copper chlorophyllin sodium, glycerophosphate, chlorophyll, sodium chloride, callopeptide, allantoin, carbazochrome, potassium nitrate, palatinit, etc. can be blended alone or in combination of two or more.

Also, alcohols, silicone, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, plastibase, etc. can be added as bases.

In this specification, the term "comprising" includes "consisting essentially of" and "consisting of." In addition, the present disclosure encompasses any and all combinations of the constituent elements described in this specification.

Furthermore, the various characteristics (properties, structures, functions, etc.) described for each embodiment of the present disclosure above may be combined in any way to identify the subject matter encompassed by the present disclosure. In other words, the present disclosure encompasses all subject matter consisting of any combination of the combinable characteristics described in this specification.

The contents of this disclosure will be explained in detail using the following experimental examples. However, this disclosure is in no way limited to these. Unless otherwise specified, experiments below were conducted under atmospheric pressure and room temperature conditions. Furthermore, unless otherwise specified, "%" means "(weight/volume)%."

Inhibition of neutrophil collagenase activity 1. Each material (alkyldiaminoethylglycine hydrochloride, dodecylpyridinium chloride, cocoyl arginine ethylpyrrolidone carboxylic acid, triclosan, sodium lauroyl sarcosine) was dissolved in a solvent (buffer included in the kit + 5% DMSO) to an appropriate concentration.
2. Neutrophil collagenase activity was evaluated using an MMP-8 fluorimetric drug discovery kit (Enzo Life Sciences, Inc., BML-AK415-0001). Specifically, 20 μl of each material solution prepared in 1 above, 20 μl of 200-fold diluted MMP-8, and 50 μl of the buffer provided with the kit were mixed and incubated at 37°C for 60 minutes. A positive control sample was prepared by adding an equal volume of buffer without adding any material, and a sample was prepared by adding an equal volume of buffer without adding MMP-8 to measure the background of the substrate alone.
3. Fluorescent substrate (BML-P126-9090) was dissolved and diluted 10-fold with buffer (concentration: 40 μM). 10 μl of diluted fluorescent substrate was added, and after 20 minutes of reaction at 37°C, the fluorescence intensity was measured at excitation light of 328 nm and emission light of 420 nm. The fluorescence intensity of the positive control sample minus the fluorescence intensity of the substrate alone as background (NC) was set to 100%, and the enzyme activity was calculated as the ratio of the fluorescence intensity of each sample with added material solution minus the fluorescence intensity of the substrate alone as background.

Figure 1 shows the results of measuring the neutrophil collagenase (MMP-8) activity of each material. The concentration on the horizontal axis indicates the concentration of each material when adjusted as described in step 1 above, which is five times higher than the concentration in the reaction system described in step 3 above. This is because when oral compositions such as toothpaste are used, they are diluted by saliva, etc., so the evaluation was conducted based on the actual conditions under which they would be used in the oral cavity.

As shown in Figure 1, alkyldiaminoethylglycine hydrochloride was confirmed to have inhibitory activity against neutrophil collagenase.

Figure 2 shows the results of measuring the neutrophil collagenase activity of alkyldiaminoethylglycine hydrochloride. The horizontal axis shows the concentration of alkyldiaminoethylglycine hydrochloride when adjusted as described above in 1.

As shown in Figure 2, alkyldiaminoethylglycine hydrochloride exhibited a neutrophil collagenase inhibitory effect of 50% or more at concentrations of 0.0031% or more, and 80% or more at concentrations of 0.0125% or more.

Claims (6)

A composition for inhibiting periodontal pocket formation and/or deepening, containing alkyldiaminoethylglycine and/or its salt as the active ingredient. A composition for inhibiting gingival collagen degradation, containing alkyldiaminoethylglycine and/or its salt as the active ingredient. A composition for inhibiting neutrophil collagenase activity, containing alkyldiaminoethylglycine and/or its salt as the active ingredient. The composition according to any one of claims 1 to 3, which is used for at least one purpose selected from the group consisting of inhibiting gingival recession, inhibiting periodontal tissue destruction, inhibiting attachment loss, and inhibiting gum recession. The composition according to any one of claims 1 to 4, wherein the total content of alkyldiaminoethylglycine and/or its salts in the composition is 0.002% by mass or more. The composition according to any one of claims 1 to 5, which is an oral composition.