AU2021103260A4 - A novel bacteriostatic oral rinse and the preparation method thereof - Google Patents

Abstract

The present invention provides a novel bacteriostatic oral rinse, which comprises bacteriostatic ingredients and substrate solution, wherein the bacteriostatic ingredients are lysozyme and cetylpyridinium chloride; and the substrate solution includes any one or more of humectants, flavoring agents, emulsifiers, flavors and chelating agents. The present invention also provides a preparation method of the novel bacteriostatic oral rinse, which includes the steps of preparing the substrate solution, adding the bacteriostatic ingredients to the substrate solution, mixing evenly, adjusting the pH value and filtering. The novel bacteriostatic oral rinse of the present invention has natural bacteriostatic active ingredients with good antibacterial effect, it has the advantages of high safety, mild composition and no harm to oral mucosa, and it is suitable for long-term daily use and maintenance of oral hygiene. 2110217CN-AU

Description

A Novel Bacteriostatic Oral Rinse and the Preparation Method Thereof

Technical Field

The present invention relates to the technical field of oral care products, and in particular relates to a novel bacteriostatic oral rinse and the preparation method thereof.

Background of the Invention

Oral rinses have been popular in developed European and American countries for many years. More and more people begin to pay attention to oral care as society develops, and oral rinses have entered ordinary families. The main functions of oral rinses are: (1) cleaning oral cavity, preventing or eliminating halitosis; (2) inhibiting dental caries; (3) helping to remove or inhibit dental plaque formation; (4) inhibiting the formation of dental scale or calculus; (5) as a supplementary to professional dental care, etc.

Chinese patent application No. 200910045463.9 discloses a complex formulation of oral rinses. Its main raw materials are chlorine dioxide, cetylpyridinium chloride and zinc ions, and the excipients are menthol, saccharin sodium salt, ethyl alcohol and glycerinum. The mass fractions of cetylpyridinium chloride and zinc ions are 0.05-0.2% and 0.5-1% separately, and the content of chlorine dioxide is 200500mg/L. However, its main ingredients are chemical disinfectants without natural antibacterial ingredients, and it contains ethanol which will have a certain irritation to oral mucosa if used for a long time.

Chinese patent application No. 201710922534.3 discloses an oral rinse containing biolysozyme, which consists of 10-40% biological lysozyme, 0.01-2.0% modified plant gum, 0.01-0.3% EDTA disodium and deionized water. However, according to the bacteriostatic mechanism of lysozyme, the bacteriostatic effect of lysozyme on Gram-positive bacteria is strong, but the bacteriostatic effect of lysozyme on Gram-negative bacteria is relatively poor, hence the bacteriostatic effect of oral rinses with lysozyme alone is limited.

Nowadays, there are various oral rinses with different formulations and effects on the market. Many of the ingredients added in oral rinses have safety risks and may cause certain harm to human body if used for a long time.

Summary of the Invention

In view of the above problems, the present invention provides an oral rinse and the preparation

1 2110217CN-AU method thereof to overcome the above problems or at least partially solve the above problems.

Specifically, the present invention is realized through the following technical solutions:

An oral rinse comprising antibacterial ingredients and substrate solution, wherein the antibacterial ingredients are composed of 0.01%~0.5% of lysozyme and 0.01%-0.5% of cetylpyridinium chloride, based on the total weight of the oral rinse.

Optionally, the content of the lysozyme is 0.1%~0.3%.

Optionally, the content of the cetylpyridinium chloride is 0.01%~0.1%.

Optionally, the substrate solution comprises any one or more of a humectant, a flavoring agent, an emulsifier, a flavor and a chelating agent; further, the substrate solution also comprises a pigment.

Optionally, in total weight of the oral rinse, the content of the humectant is 5%~20%, the content of the flavoring agent is 0.01%-5%, the content of the emulsifier is 0.04%~0.5%, the content of the flavor is 0.01%-0.5%, the content of the chelating agent is 0.01%-0.5%, and the content of the pigment is 0 ~ 0.001%.

Optionally, the oral rinse has a neutral pH value.

Optionally, the oral rinse consists of 0.1%~0.3% of lysozyme, 0.01%~0.1% of cetylpyridinium chloride, 8%-10% of humectant, 0.02%-5% of flavoring agent, 0. 2 % of emulsifier, 0.05% of flavor, 0.05% of chelating agent, 0~ 0.0003% of pigment and balance water.

Optionally, the oral rinse consists of 0.1% of lysozyme, 0.1% of cetylpyridinium chloride, 8% of humectant, 0.03% of flavoring agent, 0. 2 % of emulsifier, 0.05% of flavor, 0.05% of chelating agent and balance water; or, 0. 2 % of lysozyme, 0.05% of cetylpyridinium chloride, 10% of humectant, 0.02% of flavouring agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent and balance water; or, 0. 3 % of lysozyme, 0.01% of cetylpyridinium chloride, 8% of humectant, 5% of flavouring

2 2110217CN-AU agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent, 0.0003% of pigment and balance water.

A method for preparing the oral rinse, comprising:

(1) dissolving the humectant, the flavouring agent and the chelating agent in water to obtain a intermediate solution;

(2) mixing the emulsifier and the flavor evenly,adding the obtained mixture into the intermediate solution and mixing them evenly to obtain the substrate solution;

(3) adding the lysozyme and the cetylpyridinium chloride into the substrate solution, mixing evenly, and replenishing with water to obtain oral rinse solution; (4) adjusting pH value of the oral rinse solution to neutral, and filtering to obtain the oral rinse.

Compared with the prior art, the novel antibacterial oral rinse and the preparation method thereof have at least the following beneficial effects:

The novel antimicrobial oral rinse of the present invention has natural antimicrobial active ingredients with good antimicrobial effect. The inhibition rate of Escherichia coli, Staphylococcus aureus, Candida albicans was more than 95%. The activity of lysozyme is stable and the raw materials are safe, mild, alcohol-free and fluorine free, and will not cause harm to oral mucosa. Therefore, the oral rinses is suitable for long-term daily use and maintenance of oral hygiene.

Detailed Description of the Invention

In order to fully understand the purpose, features and effect of the present invention, the invention is described in detail through the following specific implementation modes.Unless otherwise mentioned, the technical terms involved in present invention have the meanings generally understood by those skilled in the art.

Oral rinses

For the problem that there are various oral rinses with complex ingredients but poor antibacterial effect and poor safety on the market, the inventor of the present invention creatively provides a novel antibacterial oral rinse after depth research, which includes antibacterial ingredients and substrate solution.

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The antibacterial ingredients of the oral rinse according to the present invention are lysozyme and cetylpyridinium chloride, and the contents of lysozyme and cetylpyridinium chloride are 0.01%~0.5% and 0.01%~0.5% respectively based on the total weight of the oral rinse, and preferably the contents of lysozyme and cetylpyridinium chloride are 0.1%~0.3% and 0.01 %~0.1% respectively.

Lysozyme, also known as N-acetylmuramide glycanohydrlase, is an alkaline enzyme that can hydrolyze mucopolysaccharide in bacteria. Lysozyme mainly destroys P-1,4 glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine in cell wall, so that the insoluble mucopolysaccharide in the cell wall can be decomposed into soluble sugar, which leads to the rupture of the cell wall and the release of the contents. Lysozyme has antibacterial, anti-inflammatory, antiviral effects, and will not cause damage to oral mucosa.

In the present invention, the lysozyme can be a natural lysozyme or a recombinant human lysozyme, which can both realize the purpose of the present invention. The lysozyme used in the following examples is egg white lysozyme, which is, of course, illustrative only but not restrictive.

Cetylpyridinium chloride is a kind of antibacterial agent with broad-spectrum activity, which has the function of sterilization and disinfection. When used in oral rinses, it will not break the balance of oral flora and will not stimulate oral mucosa.

Lysozyme and cetylpyridinium chloride with the above mentioned ratio are used as antibacterial ingredients in the present invention, and they are combined with each other to play synergistic effects. As a natural bacteriostatic agent, lysozyme has strong bacteriostatic effect on Gram-positive bacteria, but relatively poor on Gram-negative bacteria. Cetylpyridinium chloride is a cationic surfactant, which can interact with anions on the cell wall to increase the permeability of cytoderm and denature membrane proteins. The present invention combines cetylpyridinium chloride with lysozyme and supplement each other to enhance the bacteriostatic and bactericidal effect.

The substrate solution of the oral rinse includes any one or more of humectant, flavoring agent, emulsifier, flavor and chelating agent. Optionally, the substrate solution also includes pigment.

The humectant can be any one or more of glycerol, pentanediol, propylene glycol and sorbitol.

The flavoring agent can be any one or more of saccharin sodium, xylitol, sorbitol, glucose, fructose, galactose and sucrose.

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The emulsifier can be hydrogenated castor oil, such as PEG40, C040, etc.

Flavor can be anyone or more of mentholum, menthol, dementholized peppermint oil. By adding flavors, the mouthfeel of the oral rinse can be optimized.

The chelating agent is, for example, EDTA-2Na. The stability of the oral rinse system and lysozyme was increased by adding the chelating agent.

Pigments can be selectively added according to the specific needs of the product, for example, they can be bright blue pigments.

Preferably, according to the total weight of the oral rinse, the contents of humectant, flavoring agent, emulsifier, flavor, chelating agent and pigment are 5%~20%, 0.01%~5%, 0.04%~0.5%, 0.01%0%0.5%, 0.01%~0.5% and 0~0.001 %, separately.

With the help of the substrate solution, which is formed by combining humectant, flavoring agent, emulsifier, flavor and chelating agent in proportion, the taste and system stability can be adjusted to achieve the purpose of effective and enduring bacteriostasis.

In a preferred embodiment, the oral rinse is composed, by weight, of 0.01%~0.5% of lysozyme, 0.01%~0.5% of cetylpyridinium chloride, 5%~20% of humectant, 0.01%-5% of flavouring agent, 0.04%~0.5% of emulsifier, 0.01%~0.5% of flavor, 0.01%~0.5% of chelating agent and balance water.

In another preferred embodiment, the oral rinse is composed, by weight, of 0.1%~0.3% of lysozyme, 0.01%-0.1% of cetylpyridinium chloride, 8%~10% of humectant, 0.02%~5% of flavouring agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent and balance water.

The present invention provides a novel bacteriostatic oral rinse by selecting lysozyme and cetylpyridinium chloride as antibacterial ingredients and combining with humectant, flavouring agent, emulsifier, flavor and chelating agent. It can significantly improve the antibacterial effect, and it does not have any side effect or cause any irritation to oral mucosa by optimizing the formula. In addition, the oral rinse system is stable and can be effective for a long time.

It should be noted that the ingredients used in the oral rinse of the present invention can be obtained through market purchase and there is no special requirements for them.

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The preparation method of the oral rinse

The oral rinse provided by the present invention can be prepared according to the following steps:

(1) According to the above formula, humectant, flavoring agent and chelating agent are added into a proper amount of water and stirred until fully dissolved. The obtained mixture is heated at a temperature of 70-100°C for 15~40 minutes, and then cooled down to room temperature so as to obtain an intermediate solution.

(2) According to the above formula, emulsifier and flavor are mixed evently, heated to 4070°C and stirred for 3-10 minutes, and then added into the intermediate solution obtained in step (1) so as to obtain a substrate solution.

(3) According to the above formula, lysozyme and cetylpyridinium chloride are added into a proper amount of water and fully dissolved, and then the obtained mixture is added into the substrate solution obtained in step (2) to mix evenly. Alternatively, according to the above formula, lysozyme and cetylpyridinium chloride are directly added into the substrate solution obtained in step (2) to mix evenly. Subsequently, water is supplemented to obtain an oral rinse solution.

(4) pH value of the oral rinse solution is adjusted to neutral range such as pH 6-8, by using sodium hydroxide or disodium hydrogen phosphate, and then the oral rinse solution is filterred with a membrane of 0.22-0.65p m, e.g. a membrane of 0.45p m, to obtain the oral rinse of the present invention.

Examples

The present invention is further described by examples, but is not limited to the scope of the stated examples. In the following examples, the experimental methods without specific conditions are selected according to the conventional methods and conditions, or according to the commercial specifications. The raw materials involved in the following examples are all obtained from conventional market. The equipments involved in the following examples are conventional devices in the field.

Example 1

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The oral rinse of example 1 is composed by weight of:

Lysozyme 0.1%

Cetylpyridinium Chloride 0.1%

Glycerinum 5%

Propylene glycol 3%

Saccharin sodium 0.03%

EDTA-2Na 0.05%

Mentholum 0.05%

PEG-40 Hydrogenated Castor Oil 0.2%

Water balance to 100%

The oral rinse of example 1 is prepared as follows:

(1) According to the above formula, raw materials were weighted. Glycerinum, propylene glycol, saccharin sodium and EDTA-2Na were added into a proper amount of water and stirred until fully dissolved. The obtained mixture was heated at a temperature of 80°C for 30 minutes, and then cooled down to room temperature so as to obtain an intermediate solution.

(2) According to the above formula, mentholum and PEG-40 hydrogenated castor oil were mixed evently, heated to 50°C and stirred for 5 minutes, and then added into the intermediate solution so as to obtain a substrate solution.

(3) Lysozyme and cetylpyridinium chloride were added into a proper amount of water and fully dissolved. Subsequently, the obtained mixture was added into the substrate solution and fully mixed.

(4) pH value of the solution obtained in step (3) was adjusted to neutral pH by using disodium hydrogen phosphate, and then the solution is filterred with a membrane of 0.45p m to obtain the oral rinse.

Example 2

The oral rinse of example 2 is composed by weight of:

Lysozyme 0.2%

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Cetylpyridinium chloride 0.05%

Glycerinum 5%

Propanediol 5%

Saccharin sodium 0.02%

EDTA-2Na 0.05%

Mentholum 0.05%

PEG-40 Hydrogenated Castor Oil 0.2%

Water balance to 100%

The oral rinse of example 2 is prepared as follows:

(1) According to the above formula, raw materials were weighted. Glycerinum, propanediol, saccharin sodium and EDTA-2Na were added into a proper amount of water and stirred until fully dissolved. The obtained mixture was heated at a temperature of 80°C for 30 minutes, and then cooled down to room temperature so as to obtain an intermediate solution.

(2) According to the above formula, mentholum and PEG-40 hydrogenated castor oil were mixed evently, heated to 60°C and stirred for 8 minutes, and then added into the intermediate solution so as to obtain a substrate solution.

(3) Lysozyme and cetylpyridinium chloride were added into a proper amount of water and fully dissolved. Subsequently, the obtained mixture was added into the substrate solution and fully mixed.

(4) pH value of the solution obtained in step (3) was adjusted to neutral pH by using disodium hydrogen phosphate, and then the solution is filterred with a membrane of 0.45p m to obtain the oral rinse.

Example 3

The oral rinse of example 3 is composed by weight of:

Lysozyme 0.3%

Cetylpyridinium chloride 0.01%

Glycerinum 8%

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Xylitol 5%

EDTA-2Na 0.05%

Mentholum 0.05%

PEG-40 Hydrogenated Castor Oil 0.2%

Brilliant Blue 0.0003% Water balance to 100%

The oral rinse of example 3 is prepared as follows:

(1) According to the above formula, raw materials were weighted. Glycerinum, xylitol, EDTA-2Na and brilliant blue mother liquor were added into a proper amount of water and stirred until fully dissolved. The obtained mixture was heated at a temperature of 80°C for 30 minutes, and then cooled down to room temperature so as to obtain an intermediate solution.

(2) According to the above formula, mentholum and PEG-40 hydrogenated castor oil were mixed evently, heated to 70°C and stirred for 10 minutes, and then added into the intermediate solution so as to obtain a substrate solution.

(3) Lysozyme and cetylpyridinium chloride were added into a proper amount of water and fully dissolved. Subsequently, the obtained mixture was added into the substrate solution and fully mixed.

(4) pH value of the solution obtained in step (3) was adjusted to neutral pH by using disodium hydrogen phosphate, and then the solution is filterred with a membrane of 0.45p m to obtain the oral rinse.

Example 4: Safety Evaluation

In this example, the oral rinse of example 1 is taken as a sample for acute eye irritation test to verify its safety.

Three New Zealand rabbits, weighing 2.0-2.5kg, were used in the experiment according to the method specified in "Technical StandardforDisinfection" issued by the Ministry of health in 2002. The rabbits were raised in the normal environment. Rabbit eyes were checked before experiment, and the abnormal ones should not be used for test.

0.1ml of the oral rinse of example 1 was sucked and dripped into conjunctival sac of the rabbit

9 2110217CN-AU and the rabbit eye was passively closed for 4 seconds. After 30 seconds, the rabbit eye was washed with normal saline. The other eye of the rabbit was dripped with normal saline as control. The injury and recovery of conjunctiva, comea and iris were observed by visual inspection at 1h, 24 h, 48 h, 72 h, 7 d, 14 d and 21 d. Results: 1 h, 24 h, 48 h, 72 h, 7 d, 14 d and 21 d after dripping the oral rinse into conjunctival sac of the rabbit, no obvious abnormality was found by visual inspection, which indicates that the oral rinse of example 1 has no acute eye irritation and has high safety.

The same method was applied to test the safety of oral rinses in example 2 and example 3, and the same results were obtained as in example 1.

Example 5: Detection of Bacteriostasis Rate

The test was conducted according to C4 in GB 15979-2002 Standardfor Disposable Sanitary Products.

Escherichia coli, Staphylococcus aureus and Candida albicans were cultured for 24 h to obtain the cultures, which were subsequently prepared into bacterial suspensions (the required concentration was: 100 [ bacterial suspension was dropped into 5mL sample solution, and 4 the number of recovered bacteria was x10 ~9x104cfu/mL). Three tubes of the samples (5mL) and one tube of control were divided into four groups and placed into four sterilized plates. 100 1 above bacterial suspensions were dropped into each tube of the samples and the control respectively, followed by mixing evenly and timing. After reacting for 10min, the samples (0.5mL) were transferred by sterile forceps to the tubes containing 5mL PBS, separately, followed by fully mixing and appropriate dilution.0.5 mL liquids which were sucked from 2 to 3 selected dilution degrees were placed in two plates, separately. 15mL of nutrient AGAR medium (bacteria) or Scharbella AGAR medium (yeast) which cooled to 40~45°C was poured to each of the plates, followed by mixing evenly. The plates were turned over after AGAR solidification, and incubated at 35°C2°C for 48h (bacteria) or 72h(yeast), and finally colony counting was performed.

The experiment was repeated for 3 times, and the sterilization rate was calculated according to the following formula: X= (A- B)/A x 100% wherein X --- bacteriostatic rate, %; A --- average colony number of the control; B --- average colony number of the sample.

Evaluation criteria: antibacterial rate>50%~90%, the product has antibacterial effect,

10 2110217CN-AU antibacterial rate > 90%, the product has strong antibacterial effect.

The oral rinses of examples 1, 2 and 3 were tested respectively.

The results are shown as follows:

Strain name Oral rinse in example 1 Oral rinse in example 2 Oral rinse in example 3

Escherichia coli 98% 96% 99% 95% 96% 98% 97% 100% 99%

Staphylococcus 97% 97% 100% 99% 99% 96% 99% 98% 98%

aureus

Candida albicans 99% 98% 98% 96% 97% 96% 99% 99% 97%

It can be seen from the above experimental results that the oral rinses of the present invention have strong antibacterial effect, and the maximum bacteriostasis rate of the oral rinses to Escherichia coli, Staphylococcus aureus and Candida albicans can reach 100%.

The above examples are the preferable implementation modes of the present invention, but not limited by the above examples. Any other alternative method, modification, combination, change, simplification, etc. which is not deviated from the spirit essence and principle of the present invention shall be equivalent replacement method, which shall be included in the protection scope of the present invention.

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Claims (13)

Claims

1. An oral rinse comprising antibacterial ingredients and substrate solution, wherein the antibacterial ingredients are composed of 0.01%~0.5% of lysozyme and 0.01%~0.5% of cetylpyridinium chloride, based on the total weight of the oral rinse.

2. The oral rinse according to claim 1, characterized in that, the content of the lysozyme is 0.1%-0.3%.

3. The oral rinse according to claim 1, characterized in that, the content of the cetylpyridinium chloride is 0.0 1%~0.1 %.

4. The oral rinse according to claim 1, characterized in that, the substrate solution comprises any one or more of a humectant, a flavoring agent, an emulsifier, a flavor and a chelating agent; further, the substrate solution also comprises a pigment.

5. The oral rinse according to claim 4, characterized in that, in total weight of the oral rinse, the content of the humectant is 5%~20%, the content of the flavoring agent is 0.01%~5%, the content of the emulsifier is 0.04%~0.5%, the content of the flavor is 0.01%~0.5%, the content of the chelating agent is 0.01%~0.5%, and the content of the pigment is 0 ~ 0.001%.

6. The oral rinse according to claim 1, characterized in that, the oral rinse has a neutral pH value.

7. The oral rinse according to any one of claims 1 to 6, characterized in that, the oral rinse consists of 0.1%~0.3% of lysozyme, 0.01%~0.1% of cetylpyridinium chloride, 8%~10% of humectant, 0.02%~5% of flavoring agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent, 0- 0.0003% of pigment and balance water.

8. The oral rinse according to claim 7, characterized in that, the oral rinse consists of 0.1% of lysozyme, 0.1% of cetylpyridinium chloride, 8% of humectant, 0.03% of flavoring agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent and balance water; or, 0. 2 % of lysozyme, 0.05% of cetylpyridinium chloride, 10% of humectant, 0.02% of flavouring agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent and balance

12 2110217CN-AU water; or, 0.3% of lysozyme, 0.01% of cetylpyridinium chloride, 8% of humectant, 5% of flavouring agent, 0.2% of emulsifier, 0.05% of flavor, 0.05% of chelating agent, 0.0003% of pigment and balance water.

9. A method for preparing the oral rinse of any one of claims 1 to 8, comprising:

(1) dissolving the humectant, the flavouring agent and the chelating agent in water to obtain a intermediate solution;

(2) mixing the emulsifier and the flavor evenly,adding the obtained mixture into the intermediate solution and mixing them evenly to obtain the substrate solution;

(3) adding the lysozyme and the cetylpyridinium chloride into the substrate solution, mixing evenly, and replenishing with water to obtain oral rinse solution; (4) adjusting pH value of the oral rinse solution to neutral, and filtering to obtain the oral rinse.

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