JP2003192608A - Agent for preventing attachment of dental caries bacteria - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an agent for preventing the attachment of dental caries bacteria, specifically inhibiting the attachment of Streptococcus mutans in the oral cavity by a peptide derived from a saliva protein and exhibiting a high effectiveness for the prevention of the dental caries. <P>SOLUTION: This agent for preventing the attachment of the dental caries bacteria contains at least one of peptides having ≤15 amino acid residues and an LYP residue at its N-terminal and/or a YTF residue at its C-terminal, and its derivatives as an active ingredient. <P>COPYRIGHT: (C)2003,JPO

Description

Description: TECHNICAL FIELD [0001] The present invention relates to a specific peptide which specifically inhibits the colonization of mutans streptococcus, one of the cariogenic bacteria, in the oral cavity. In addition, the present invention relates to an agent for preventing cariogenic bacteria from being able to prevent or treat caries. [0002] Caries are caused by pathogenic bacteria, mainly Streptococcus mutans, which adhere to and adhere to the oral cavity, and the acid produced by these bacteria is enamel. Is caused by demineralization. Therefore, to prevent dental caries, it is effective to remove the causative bacteria using a bactericide. However, fungicides act non-specifically on bacteria, and from the sense of resistance to indiscriminately destroying the bacterial flora in the oral cavity, a method that specifically eliminates only the causative bacterium, Streptococcus mutans, is desired. For example, antibodies obtained by immunizing animals or poultry with Streptococcus mutans have been proposed. However, antibodies are high molecular compounds and are less likely to act on pathogenic bacteria living inside plaque, and it is desired to develop a more effective specific adhesion inhibitor. Means for Solving the Problems and Embodiments of the Invention The present invention has been made in order to meet the above-mentioned demands, and various studies have been made on an antibacterial agent specific to Streptococcus mutans. However, the present inventors have found that a peptide having an LYP residue at the N-terminus and a YTF residue at the C-terminus and having no more than 15 amino acids specifically prevents adhesion of Streptococcus mutans to the oral cavity. It was completed. Hereinafter, the present invention will be described in detail. The anti-caries agent of the present invention comprises a peptide having 15 or less, preferably 5 to 9 amino acids having LYP residues at the N-terminal and / or YTF residues at the C-terminal, and derivatives thereof. At least one is an active ingredient. The abbreviations for the constituent amino acids of the peptide and the protective groups thereof used in the present specification are those commonly used in the field of peptides. For example, L is leucine, Y is tyrosine, P is proline, Q is glutamine,
T represents threonine, and F represents phenylalanine. The mutans bacterium has an adhesion factor (PAc, P1, SAI / II, etc.) specific to the surface of the bacterium and specifically binds to saliva components constituting a pellicle formed on the tooth surface. Is known. Thus, a method has been proposed in which the active site peptide of this specific adhesin is used to competitively suppress mutans-specific adhesion.
ature Biotechnology 17,42
-47, 1999). On the other hand, statherin is a salivary protein consisting of 43 amino acids, and has been reported to have an effect of inhibiting precipitation of calcium phosphate in saliva, but is considered not to function as an adhesin for mutans bacteria. However, surprisingly, the C-terminus of this statherin (2
(9-43 residues) side, the effect of suppressing the specific adhesion of mutans bacteria was found. This statherin C-terminal (29
-43 residues) Both ends of the peptide, ie, LY at the N-terminus
Although an effect is also recognized on a peptide containing a P residue and a C-terminal YTF residue, in the present invention, particularly, LYPQPYQ
PQ and PQYQQYTF are desirable, and furthermore, LYPQYTF in which the two (LYP and YTF) are combined is desirable. [0008] The peptide can be easily synthesized by a solid phase method commonly used for peptide synthesis. For example, it can be synthesized according to the method of Merrifeed et al. Described in Journal of Chemical Society, Vol. 85, pp. 2149-2154 (1963). It can also be synthesized using a commercially available peptide synthesizer or the like. In addition, the gene encoding the amino acid sequence of the peptide of the present invention can be produced in large amounts in microbial cells, plant cells, and animal cells using genetic engineering techniques. The obtained crude synthetic peptide can be further purified by means commonly used for protein / peptide purification, such as gel filtration, reverse phase HPLC, and ion exchange column purification. In the present invention, these peptides can be used alone or in combination of two or more. [0010] The anti-cariogenic bacteria adhesion inhibitor of the present invention can be used by blending it with various oral care agents (oral compositions).
Is preferably 0.0001 to 5% (mass percentage, the same applies hereinafter) of the whole composition, particularly preferably 0.001 to 1.0%. In the oral composition of the present invention, these peptides can be used alone or in combination of two or more, and the amount of the peptide in the composition is 0.0001.
To 5%, particularly preferably 0.001 to 1.0%. The caries-causing bacteria adhesion inhibitor according to the present invention comprises:
Thus, various oral care agents, for example, oral solution, oral pasta, toothpaste, mouthwash, oral troche, tablet,
Applicable to chewing gum and the like. Each composition can be prepared by a usual method using other components according to the characteristics thereof, as long as the effects of the present invention are not impaired. Optional components include, for example, abrasives such as calcium hydrogen phosphate dihydrate, calcium hydrogen phosphate anhydride, calcium carbonate, aluminum hydroxide, aluminum oxide, and silicic acid, and viscous agents such as glycerin, sorbite, and propylene glycol. Agents, binders such as sodium carboxymethylcellulose, carrageenan, xanthan gum, surfactants such as sodium lauryl sulfate, sweeteners such as saccharin, other preservatives, flavors, coloring agents, pH adjusters, excipients, and various medicinal ingredients Etc. can be blended. EFFECT OF THE INVENTION The bacterial adhesion inhibitor of the present invention, which uses a peptide derived from salivary protein, specifically inhibits the colonization of Streptococcus mutans in the oral cavity and is very effective in preventing caries. Next, the effects of the present invention will be described specifically with reference to examples, but the present invention is not limited to the following examples. Example: Bacterial Adhesion Inhibition Experiment As a tooth model, hydroxyapatite beads (manufactured by Asahi Kogaku Co., Ltd.) were reacted with human filtered saliva for 60 minutes, and further, an albumin solution (5
mg / ml) for 30 minutes. After washing, K
Cl buffer (50 mM KCl, 1 mM KH ₂ PO ₄ ,
1 mM CaCl ₂ , 0.1 mM MgCl ₂ , pH 7)
[H ³ ] -labeled bacterial solution (about 10 ⁷ ce)
ls) and the sample solution were added and reacted for 60 minutes. The bacterium is Streptococcus mutans
coccus mutans), Streptococcus gordonii (Streptococ)
cus gordonii, Actinomyces naeslund
dii) was used. After washing, the apatite beads were transferred to vials, and their radioactivity was measured with a liquid scintillation counter to determine the amount of adhered bacteria. The adhesion inhibition rate is
It was calculated according to the following equation. Table 1 shows the results. ## EQU1 ## From the results shown in Table 1, each peptide does not act on indigenous bacteria, but shows an inhibitory effect on mutans bacteria only to salivary-coated hydroxyapatite beads, which is equal to or more than that of an antibody against mutans bacteria adhesion factor. It can be seen that the effect is obtained. [Table 1] In the following, prescription examples are shown. [Formulation Example 1] Oral solution Sorbit 10.0% Ethanol 5.0 Polyoxyethylene (60 mol) hydrogenated castor oil 0.1 Sucrose monopalmitate 0.2 Sodium lauryl sulfate 0.05 Saccharin sodium 0.2 LYPQYTF 02 Perfume 0.6 Water Balance 100.0% [Formulation Example 2] Oral Pasta Cetanol 5.0% Squalane 20.0 Precipitable Silica 5.0 Polyoxyethylene (40 mol) Hydrogenated Castor Oil 0.1 Sorbitan monooleate 1.0 Sodium lauryl sulfate 0.2 Saccharin sodium 0.6 LYPQYTF 0.05 Fragrance 0.6 Water balance 100.0% [Prescription example 3] Dentifrice Silicic anhydride 15.0 Lauryl sulfate Sodium 1.0 Sorbitol 20.0 Xylitol 5.0 Xanthan gum 0.5 Sodium 0.5 Sodium saccharin 0.1 Sodium monofluorophosphate alginic 0.1 LYPQYTF 0.01 Perfume 1.0 Purified water Balance 100.0% [0022] Formulation Example 4 mouthwashes Ethanol 10.0 LYPQYTF 0.1 Polyoxyethylene (60 mol) hydrogenated castor oil 0.5 Sorbitan sodium saccharin 0.2 Sodium fluoride 0.1 Xylitol 5.0 Preservative and flavor 0.8 Distilled water Balance 100.0% [Formulation Example 5] Gum Gum base 20.0 Flavor 1.0 Sugar syrup 20.0 Sorbitol 20.0 Xylitol 10.0 Powdered sugar 20.0 Trisodium citrate 1.5 Sodium monofluorophosphate 0.5 LYPQYTF 0.1 water balance 100. %

Continuation of front page    F term (reference) 4B014 GB13 GL09                 4C083 AA071 AA072 AB172 AB472                       AC102 AC132 AC432 AC782                       AC862 AC902 AD302 AD352                       AD411 AD412 BB55 CC41                       DD31 EE32 FF01                 4C084 AA01 AA02 BA01 BA08 BA17                       BA18 CA18 CA35 DC50 MA57                       NA14 ZA671 ZC412                 4H045 AA30 BA14 BA15 BA17 CA40                       EA20 FA10 FA74

Claims (1)

Claims: 1. A caries containing, as an active ingredient, at least one of a peptide having 15 or less amino acid residues having an LYP residue at the N-terminus and / or a YTF residue at the C-terminus and a derivative thereof. Causative bacteria adhesion inhibitor.