JP4409229B2 - Coaggregation inhibitor - Google Patents

Description

  The present invention relates to a coaggregation inhibitor that suppresses coaggregation of periodontal disease-related bacteria in the oral cavity.

  Periodontal disease is an infectious disease of the oral cavity that is caused by the adhesion and establishment of pathogenic bacteria on the tooth surface. The mechanism of colonization of periodontal disease-related bacteria on the tooth surface is as follows. First, initial colonization bacteria such as Streptococcus oralis, Streptococcus sangais, Streptococcus gordonii, Actinomyces naeslandi, etc. Adsorbs. These early colonization bacteria co-aggregate with each other as they grow, and start to form plaque. Next, with the maturation of the plaque, the microbial flora transitions from facultative anaerobes to obligate anaerobes, and obligate anaerobes typified by Fusobacterium nucleatum co-aggregate with early-fixing bacteria. And it is thought that periodontal disease related bacteria, such as Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia, co-aggregate and settle in the Fusobacterium nucleatum.

  Coaggregation is caused by non-specific electrostatic interactions between bacteria, lectin-receptor-type interactions, adhesion by sticky polysaccharide synthesis, and non-specific hydrophobic interactions. It plays a particularly important role in the settlement.

As a means of preventing oral infections, it has hitherto been considered to be effective in inhibiting the colonization of pathogenic bacteria on the tooth surface. For example, coaggregation of oral bacteria using specific amino sugars Inhibits plaque formation and suppresses plaque formation (see, for example, Patent Document 1); Inhibiting plaque construction using oligosaccharides isolated from sangis (see, for example, Patent Document 2), and inhibiting bacterial aggregation using xanthan gum, gum tragacanth, guar gum, etc. (for example, Patent Document) 3), etc. have been reported. In addition, it has been reported that an antibacterial preparation comprising lysine or a derivative thereof, a compound exhibiting antibacterial activity, and a surfactant exhibits an antibacterial effect on an aggregate or mass of microorganisms (see, for example, Patent Document 4).
JP-A-6-24948 US Pat. No. 7,349,772 US Pat. No. 4,855,128 JP-A-8-151325

  The present invention prevents the adhesion and colonization of periodontal disease-related bacteria on the tooth surface by suppressing the coaggregation of periodontal disease related bacteria on the tooth surface, and is useful for the prevention and improvement of periodontal disease related diseases. An object is to provide an oral preparation.

  The present inventors focused on coaggregation of oral bacteria involved in plaque formation and examined components that suppress coaggregation. As a result, lysine, histidine, or a salt thereof was adhered to Fusobacterium nucleatum adhering to early colonized bacteria and teeth. It has been found that it has a coaggregation-inhibiting action of periodontal disease-related bacteria, prevents adhesion and colonization of periodontal disease-related bacteria on the tooth surface, and is useful for prevention and improvement of periodontal disease-related diseases.

  That is, the present invention provides a coaggregation inhibitor of Fusobacterium nucleatum and periodontal disease-related bacteria containing lysine, histidine or a salt or derivative thereof as an active ingredient.

  The coaggregation inhibitor of the present invention suppresses coaggregation of Fusobacterium nucleatum and periodontal disease-related bacteria, and can prevent adhesion of periodontal disease-related bacteria to the tooth surface. Therefore, by using this, periodontal disease-related diseases such as periodontal disease and alveolar pyorrhea can be prevented and improved.

  The lysine and histidine which are the active ingredients of the coaggregation inhibitor of the present invention may be any of L-form, D-form and DL-form, but is preferably L-form. Examples of lysine or histidine salts include hydrochlorides, sulfates, nitrates, phosphates, acetates, etc. Of these, hydrochlorides are preferred.

Such lysine or histidine or a salt thereof is preferably L-lysine, a salt of L-lysine or L-histidine, and particularly preferably L-lysine hydrochloride.
In addition, lysine or histidine or a salt thereof can be chemically synthesized by a conventional method, or a commercially available product may be used.

Lysine or histidine or a salt thereof inhibits co-aggregation of Fusobacterium nucleatum and periodontal disease-related bacteria, as shown in Examples below.
Fusobacterium nucleatum (Fusobacterium nucleatum), in plaque formation, initial fixing bacteria adhere and aggregate on the tooth surface covered by a pellicle (e.g., Streptococcus oralis (Streptococcusoralis), Streptococcus Sangaisu (Streptococcus sanguis), Streptococcus Godonyi (Streptococcusgordonii), It is an obligate anaerobic bacterium that adheres to Actinomyces naeslundii . It is thought that the adhesion and colonization of periodontal disease-related bacteria to the tooth surface is caused by co-aggregation of this Fusobacterium nucleatum with periodontal disease-related bacteria, so that Fusobacterium nucleatum and periodontal disease-related bacteria If coaggregation is suppressed, it becomes possible to prevent periodontal disease-related bacteria from adhering to and fixing on the tooth surface.

Here, periodontal disease-related bacteria include, for example, Actinobacillus actinomycetemcomitans and other Actinobacillus bacteria, Porphyromonas gingivalis and other Porphyromonas genus bacteria, Prebotella Prevotella bacteria other such Intermedia (Prevotellaintermedia), Bacteroides Fosaisasu (Bacteroides fosythus), Treponema Dentikora (Treponemadenticola), and the like.

  Thus, lysine or histidine or its salt prevents periodontal disease-related bacteria from sticking to the tooth surface, thus preventing and improving periodontal disease-related diseases such as periodontal disease and alveolar pyorrhea. It is useful as a preparation for oral cavity that exhibits the effect.

  That is, the coaggregation inhibitor of the present invention is a component used in ordinary oral compositions, such as foaming agents, foaming aids, surfactants, abrasives, bulking agents, sweeteners, preservatives, medicinal ingredients, pH. Toothpaste, liquid toothpaste, liquid toothpaste, moisturized toothpaste, mouthwash, mouth spray, antibacterial / improving effect on periodontal disease-related diseases by appropriately blending regulators, adhesives, pigments, pigments, fragrances Chewing gum can be used.

  Here, examples of the medicinal component include fungicides such as sodium fucca, triclosan, chlorhexidine and benzethonium chloride, enzyme inhibitors such as polyphenols (glucosyltransferase inhibitory action), and anti-inflammatory agents such as indomethacin and dipotassium glycyrrhizinate. Ordinarily used as oral preparations can be used.

  Sweeteners include oligosaccharides or sugar alcohols such as xylo-oligosaccharides, fructooligosaccharides, galactooligosaccharides, soybean oligosaccharides, isomaltoligosaccharides, dairy oligosaccharides, lactulose, raffinose, trehalose, glucosyl sucrose, maltosilche. Claus, palatinose, maltitol, erythritol, reduced palatinose, xylitol and the like are preferable examples.

  In addition, the content of lysine or histidine or a salt thereof in such a coaggregation inhibitor is preferably 0.001 to 10% by weight, particularly preferably 0.01 to 5% by weight from the viewpoint of production cost. .

<Coaggregation inhibition test>
(1) Strain used Periodontal disease related bacteria ( Porphyromonas gingivalis ATCC33277 (Pg); Prevotella intermedia ATCC25611 (Pi); Actinobacillus actinomycetemcomitans ATCC33384 (Aa)) and oral bacteria ( Fusobacterium nucleatum ATCC25586 (Fn)) were used.

(2) Culture and washing Periodontal disease-related bacteria are K1, H Shedler broth (Becton Dickinson), Fusobacterium nucleatum (Fn) is GAM bouillon (Nissui Pharmaceutical Co., Ltd.), 37 ° C, 10% CO ₂ , 10 The cells were cultured from the logarithmic growth phase to the stationary phase under the conditions of% H ₂ and 80% N ₂ , and collected by centrifugation at 5000 rpm for 5 minutes. Thereafter, the mixture was washed three times with PBS buffer (144 mg / L KH ₂ PO ₄ , 9 g / L NaCl, 795 mg / L Na ₂ HPO ₄ , pH 7.4) (5000 rpm, 5 minutes), and adjusted to 1.0 at OD 600 nm.

(3) Tube method When evaluating coaggregation-inhibiting materials, add 200 μL to the test tube so that the final concentration of the material is 100 mM, then dispense 600 μL of Fusobacterium nucleatum (Fn) and add 600 μL of periodontal disease-related bacteria. After mixing and stirring, the mixture was stored at 4 ° C. for 19 hours, and then the coaggregation reaction was visually confirmed.

(4) Coaggregation measurement method using 96-well microplate
After adding 20 μL of various materials to a 96-well microplate so that the final concentration was 100 mM, 60 μL of Fusobacterium nucleatum (Fn) was dispensed and stirred, and 60 μL of periodontal disease related bacteria was added. Then, using a microplate reader (Molecular Device), measure the absorbance (OD600nm) for 30 minutes at 1 minute intervals, calculate the absorbance change rate Vmax (mOD / min), and calculate the relative coaggregation rate. Aggregation reaction was measured.

Example 1
Using L-histidine (Wako Pure Chemical Industries) or L-lysine hydrochloride (Wako Pure Chemical Industries) and water, L-arginine (Wako Pure Chemical Industries) or D-galactose (Wako Pure Chemical Industries) The coaggregation inhibitory effect of Fusobacterium nucleatum (Fn), which is a bacterium, and Prevotella intermedia (Pi), which is a periodontal disease-related bacterium, was measured by a 96-well microplate method. As a result, the relative co-aggregation rate was 100% for water, D-galactose, and L-arginine, but 84% for L-histidine and 20% for L-lysine hydrochloride. L-histidine or L-lysine hydrochloride The coaggregation inhibitory effect was recognized by the salt (FIG. 1). Furthermore, when the coaggregation inhibitory effect of lysine was examined by the tube method, in water, aggregates appeared at the bottom of the tube, but in L-lysine hydrochloride, aggregates did not appear. Similarly, a high coaggregation suppressing effect was observed (FIG. 2).

Example 2
Using L-lysine hydrochloride (Wako Pure Chemical Industries) and PBS and D-galactose (Wako Pure Chemical Industries) 100 mM each for comparison, oral bacteria Fusobacterium nucleatum (Fn) and periodontal disease-associated Porphyromonas gingivalis ( The coaggregation inhibitory effects of Pg) and Actinobacillus actinomycetemcomitans (Aa) were measured by the tube method. As a result, PBS and D-galactose showed aggregates at the bottom of the tube, but L-lysine hydrochloride did not show aggregates, and L-lysine hydrochloride showed a high coaggregation inhibitory effect ( 3 and 4).

FIG. 1 is a diagram showing the coaggregation inhibitory effect (microplate method) of Fn and Pi. FIG. 2 is a diagram showing the coaggregation suppressing effect (tube method) of Fn and Pi. FIG. 3 is a diagram showing the coaggregation inhibitory effect (tube method) of Fn and Pg. FIG. 4 is a diagram showing the coaggregation suppressing effect (tube method) of Fn and Aa.

Claims (1)

A co-aggregation inhibitor of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis or Prevotella intermedia , wherein the active ingredient is composed of lysine or histidine or a salt thereof.