JPS63185928A - Antimicrobial agent - Google Patents

Abstract

PURPOSE:To obtain an antimicrobial agent, containing a specific pyrazolopyrazine derivative as an active ingredient, having a powerful antimicrobial activity and excellent antimicrobial action on dental caries, periodontosis, cariogenic bacteria, etc., as well as blending characteristics and feeling of use. CONSTITUTION:The aimed substance obtained by using a 1,5-substituted-1H- pyrazolo[3,4-b]pyrazine derivative expressed by the formula [R1 is H, OH, halogen, hydrazino, group -O-R' or -NH-R' (R' is substituted phenyl, etc.); R2 is H, alkyl, benzyl or phenyl] alone or, if necessary, together with other antimicrobial agents, preservatives, etc. blending in a composition for the oral cavity, chemicals, cosmetics, etc., and preparing a pharmaceutical by a conventional method. If the compound expressed by the formula is blended in the composition for the oral cavity, the amount thereof blended is 0.00001-1wt.% based on the total amount of the ingredients. The agent is preferably used without bitterness or tooth coloring action when used in the composition for the oral cavity, e.g. tooth paste, mouthwash, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION Part 1: The present invention relates to an antibacterial agent containing a specific pyrazolopyrazine derivative as an active ingredient. The present invention relates to an antibacterial agent that has excellent antibacterial activity against bacteria and can be used as a bactericidal agent or preservative in various oral compositions, cosmetics, drugs, etc.

The following are known as antibacterial substances that have been used as disinfectants in various drugs and cosmetics.

■Antibiotics: tetracycline, penicillin, etc. ■Quaternary ammonium salts: cetylpyridinium chloride, benzalkonium chloride, etc. ■Biguanide compounds: chlorhexidine, etc. ■Phenols ■Alcohols ■Oxidizing agents: Hydrogen peroxide, especially these Among them, chlorhexidine is relatively widely used in oral product groups.

17 Problems to be Solved by ■ However, talolhexidines tend to be deactivated when they coexist with anionic surfactants, and when added to oral compositions, they have a strong bitter taste and may even stain the teeth. There is.

Therefore, there has been a demand for an antibacterial agent that not only has excellent antibacterial activity but also has excellent formulation characteristics and usability.

The present invention was developed in view of the above circumstances, and it has a strong antibacterial effect, but also has a weakened antibacterial effect.

To provide an antibacterial agent that does not become inactivated, has no bitter taste or tooth staining effect when used in the oral cavity, and can be suitably used as an oral bactericide.

Means and Action for Solving the Problems In order to achieve the above object, the present inventors have conducted extensive searches for antibacterial agents, and have found that certain compounds among pyrazolopyrazine derivatives, which are antitumor active substances, have excellent antibacterial properties. found to be active.

That is, the present applicants previously proposed that 1,5-substituted-IH-pyrazolo[3,4-b copyrazine derivatives are excellent as antitumor agents (Japanese Patent Application No. 116852/1982).
No. 61-161015), of which the following formula (1) [However, R1 is a water an alkyl group, a benzyl group, a phenyl group, or a phenyl group in which one or more hydrogen atoms of the phenyl group are substituted with a halogen atom, an alkyl group, or an alkoxy group), R2 is a hydrogen atom, an alkyl group, a benzyl group, or a phenyl group; show. The 1,5-substituted-IH-pyrazolo[3゜4-b]pyrazine derivative shown in It has excellent anti-formation ability, and unlike antibiotics, it is difficult for resistant bacteria to appear, and its antibacterial activity does not become inactive when combined with anionic surfactants, such as chlorhexidine. It has been found that even when used in the oral cavity, it does not have a bitter taste, mucosal irritation, or tooth staining effect.

Then, they discovered that by using the compound of formula (1) as an active ingredient, a high-quality antibacterial agent with strong antibacterial activity and that can be effectively used for oral sterilization was obtained, leading to the present invention. Ta.

Therefore, the present invention provides 1,5- represented by the above general formula (1).
Provided is an antibacterial agent characterized by containing a substituted-IH-pyrazolo[3,4-bl pyrazine derivative as an active ingredient.

The present invention will be explained in more detail below.

The antibacterial agent of the present invention can be used as a bactericidal agent or preservative in oral compositions, medicines, cosmetics, and the like.

In particular, by blending it into oral compositions such as drills and mouthwashes and using it in the oral cavity, it is possible to suppress oral disease-causing bacteria and effectively prevent orovaginal diseases.

As mentioned above, the antibacterial agent of the present invention has the formula (1), that is, [where R1 is a hydrogen atom, a hydroxyl group, a halogen atom, a hydrazino group, a -〇-R' group or a -NH-R' group (here, R' is an alkyl group, a benzyl group, a phenyl group, or a phenyl group in which one or more hydrogen atoms of the phenyl group are substituted with a halogen atom, an alkyl group, or an alkoxy group)
, R2 represents a hydrogen atom, an alkyl group, a benzyl group or a phenyl group. Same below. ] 1,5-substituted-L
Contains H-pyrazolo[3,4-bl pyrazine derivative as an active ingredient. In this way, by containing the 1,5-substituted-L H-pyrazolo[3,4-b copyrazine derivative of formula (1), it has strong antibacterial activity, and is particularly effective against dental caries, periodontal pathogenic bacteria, and bad breath. It effectively suppresses oral disease-causing bacteria such as causative bacteria, and also has excellent ability to inhibit plaque formation. In addition, it does not lose its activity when used in combination with anionic surfactants, etc., and does not cause bitterness or mucosal irritation even when used in the oral cavity. , it is possible to obtain an antibacterial agent that does not have a tooth-staining effect and can be suitably used as an oral-vaginal disinfectant.

Here, in formula (1), the halogen atom is CQ
, Br, etc., and the alkyl group is CnH, n, etc.

(However, n = 1 to 8) etc., but the alkoxy group is OC
Examples include n H2n+1 (where n=1 to 8).

In addition, among the compounds of formula (1), 5-alkoxy-IH-pyrazolo [where R is an alkoxy group and R2 is a hydrogen atom]
3,4-bl pyrazine derivatives have excellent antibacterial activity,
Specifically, 5-hexyloxy-IH-pyrazolo[3,
4-bl pyrazine and the like are preferably used.

The compound of formula (1) can be produced by various methods, and the production method can be appropriately selected depending on the type of compound, conditions, etc. For example, 5-alkoxy-IH-pyrazolomini 3,4-
b] Pyrazine derivatives are easily synthesized by reacting 5-alkoxy-1-vinyl-IH-pyrazolo [3゜4-b] pyrazine derivatives with permanganates such as potassium permanganate or organic peracids. be able to.

In the present invention, the compound of formula (1) can be used alone or in combination with other antibacterial agents, preservatives, etc., in oral compositions, drugs, cosmetics, etc. It can be applied to

The amount of the compound in (1) varies depending on the form of the antibacterial agent, the type of composition in which the antibacterial agent is mixed, the area of application, the method and number of applications, etc., and can vary widely depending on the severity of the symptoms. However, for example, when blended into an oral composition, the compound of formula 1) is 0.0000% of the total amount of components.
It is preferable to blend it in an amount of 1 to 1% by weight, particularly 0.001 to 0.1% by weight. It should be noted that an antibacterial agent containing the compound of formula (1) within this range does not have a bitter taste, mucosal irritation, or tooth staining effect even when applied to the oral cavity.

The antibacterial agent according to the present invention can be blended into various compositions as described above, and appropriate commonly used components can be blended into such compositions.

For example, when incorporating the antibacterial agent of the present invention into an oral composition, in the case of toothpaste, dibasic calcium phosphate, calcium carbonate, calcium pyrophosphate, insoluble sodium metaphosphate, amorphous silica, crystalline silica, Abrasives such as aluminosilicate, aluminum oxide, aluminum hydroxide, resin, sodium carboxymethyl cellulose, hydroxyethyl cellulose, alginate.

Binder such as carrageenan, gum arabic, polyvinyl alcohol, thickening agent such as polyethylene glycol, sorbitol, glycerin, propylene glycol, sodium lauryl sulfate, sodium dodecylbenzenesulfonate, hydrogenated coconut fatty acid monoglyceride monosulfate, lauryl sulfonic acetic acid. Sodium, sodium N-lauroyl sarcosinate, N-acylglutamate, surfactants such as sucrose fatty acid ester, essential oils such as peppermint and spearmint, Q-menthol,
Flavoring materials such as carvone, eugenol, anethole, etc., and sodium saccharin.

Stevioside, neohesberidyl dihydrochalcone,
It is produced by mixing ingredients such as sweeteners such as glycyrrhizin and perilartin with water and following a conventional method.

In addition, in mouthwashes and other mouthwashes, ingredients are appropriately blended depending on the properties of the product.

In addition, in the antibacterial agent of the present invention, dextranase,
Mutanase, sorbic acid, alexidine.

Hinokitiol, chlorhexidine, alkylglycine, alkyldiaminoethylglycine salt.

Active ingredients such as allantoin, ε-aminocaproic acid, tranexamic acid, azulene, vitamin E, and crude drug extracts can also be used in combination.

Effects of the Invention The antibacterial agent of the present invention is a 1,5-substituted-1H-
Contains pyrazolo[3,4-bl pyrazine derivative as an active ingredient, it has strong antibacterial activity, and has antibacterial activity and ability to inhibit dental plaque formation, especially against caries-causing, fi-periopathic, and halitosis-causing bacteria. Furthermore, the antibacterial activity of the compound of formula (1) does not become inactivated or weakened, and it has no bitter taste, mucosal irritation, or tooth staining effect, making it suitable as an antibacterial agent for the oral cavity.

Examples are shown below.

[Example 1] The antibacterial activity of the 1,5-substituted-IH-pyrazolo[3,4-b copyrazine derivative against caries-causing, periodontal disease-causing bacteria, and halitosis-causing bacteria was evaluated by the method shown below.

Test drug 5-hexyloxy-IH-pyrazolo[3,4'-
b] Pyrazine was dissolved in dimethyl sulfoxide at a concentration of 0.1% and sterilized (φo, 22p).

Test strain Seven bacterial strains of normal vaginal bacteria shown in Table 2 were tested.

In addition, the strain is Todd Hetiitt broth
(Difc.

3μ of 5% sheep skin defibrinated blood on an agar medium based on
The cells were cultured on a blood agar medium supplemented with 0.5 μg/mQ of hemin and 0.5 μg/mQ of menadione.

Antibacterial activity test drug (5-hexyloxy-IH-pyrazolo [3,
4-b] pyrazine) at a final concentration of 3.13 μg/d ~ 2
Liquid medium (3%
1150, t
inoculated under anaerobic conditions (80% N2, 10% H2, 1
The cells were cultured at 37°C (0% CO2) for 5 days. Note that the concentration of dimethyl sulfoxide in the liquid medium is 5%,
There was no effect on the growth of the test bacterial strain.

After culturing, the degree of bacterial growth is measured by turbidity (○, D, 550 nm)
The minimum inhibitory concentration (MIC) of the test drug against various bacteria was determined. Furthermore, the culture solution was spread on a blood agar medium and cultured for 5 days under the same conditions as above, and the growth of bacteria was observed to determine the minimum bactericidal concentration (MBC).

The above results are shown in Table 2.

Table 2 From the results shown in Table 2, 5-hexyloxy-L H-pyrazolo[:3.4-bl pyrazine has excellent ability to inhibit the growth of dental caries, periodontal pathogenic bacteria, and bad breath-causing bacteria. It was confirmed that

[Example 2] i of 1,5-substituted-IH-pyrazolo[3,4-bl pyrazine derivatives against Streptococcus mutans
A7 Ability to inhibit formation of scale base was evaluated by the method shown below.

Reaction solution 4- with the composition shown below was placed in a test tube, held at an angle of 30', and heated under anaerobic conditions (80%N).
, 10% H, 10% Go2) and cultured at 37°C for 5 days.

<Reaction solution composition> Streptococcus mutans culture solution - 0.1 total
4. OmQ - Streptococcus mutans strain 6715 was cultured in brain heart infusion culture solution for 24 hours, and then diluted 5 times with physiological saline.

Next, after decanting and discarding the liquid in the test tube, 0.5N sodium hydroxide solution 4-
was added and left for 1 hour. After leaving it for a while, evenly remove the plaque attached to the test tube! The amount of dental plaque was expressed as turbidity by measuring the absorbance at a wavelength of 660 nm, and the presence or absence of bacterial growth was determined from this turbidity.

The results are shown in Table 3.

Table 3 From the results in Table 3, 1,5-substituted-IH-pyrazolo[3
, '4-b] pyrazine derivative 5-hexyloxy-I
H-pyrazolo[3,4-bl pyrazine was found to have excellent plaque formation inhibiting ability.

In addition, the above-mentioned 5-hexyloxy-IH-pyrazolo[3
,4-bl pyrazine is, for example, 5-hexyloxy-1
-vinyl-IH-pyrazolo[3°4-b] It can be obtained by dissolving copyrazine in a suitable solvent, adding 5% aqueous potassium permanganate solution or metachloroperbenzoic acid, heating to reflux, and reacting.

Examples of formulations of the antibacterial agent of the present invention are shown below, but the antibacterial agent of the present invention is not limited to the following formulation examples.

[Formulation example 1] Toothpaste calcium diphosphate 50% by weight
Glycerin 2゜Sodium lauryl sulfate 1 Sodium carboxymethyl cellulose 1 Sodium saccharin
0. 1 Sodium monofluorophosphate 0.5 Ethanol
5 total
ioo, o wt% When the above dental calendar was prepared and used by five panelists twice a day after scaling for about one month, there was no bitter taste and no side effects such as mucous membrane irritation or tooth discoloration were observed. Ta.

[Formulation example 2] Mouthwash ethanol 20% by weight Sodium saccharin 0.0
5 Flavoring (cinnamon oil: menthol = 4:1) 1 Sodium monofluorophosphate 0.1 Sucrose palmitate 0.5 Purified water
Remaining total 100.0% by weight
[Formulation example 3] Chewing gum gum base 20% by weight calcium carbonate dihydrate syrup
15 Powdered sugar 30 Maltose 10 Fructose 105-hexyloxy-IH-pyrazolo 0.01 [3,4-b Copyrazine Purified water Remaining total 100.0% by weight [Formulation example 4] Lozenge Gum arabic 6% by weight Fructose 20 Glucose 20 Maltose 30 dextranase
2 Purified water balance 100.0% by weight [Formulation example 5] Oral pine surge cream white petrolatum
8% by weight stearyl alcohol 6 propylene glycol
4 dextranase
2 Polyethylene glycol 4000 2
5 Polynylene glycol 400 37 Ethanol 7 Purified water
Remaining total ioo, o wt% [Formulation example 6] Skin cream squalane 10.0 wt% Vaseline 9 Beeswax 4 Microwax 8 Isopropyl myristate 5 2-octyldodecyl myristate 10 Polyoxyethylene monostearate
4 Sorbitan monostearate 4
Propylene glycol 10 semen water
Residue

Claims (1)

[Claims] 1. The following formula (1) ▲There are mathematical formulas, chemical formulas, tables, etc.▼...(1) (However, R_1 is a hydrogen atom, a hydroxyl group, a halogen atom, a hydrazino group, -O-R' group or -NH-R' group (where R' is an alkyl group, a benzyl group, a phenyl group, or a phenyl group in which one or more hydrogen atoms of the phenyl group are substituted with a halogen atom, an alkyl group, or an alkoxy group) , R_1 represents a hydrogen atom, an alkyl group, a benzyl group, or a phenyl group. agent.