RU2745296C1 - Method for producing bactericidal agent based on hydrolysates of coconut oil for oral cavity care - Google Patents

Abstract

FIELD: cosmetic and pharmaceutical industry.

SUBSTANCE: invention relates to the cosmetic and pharmaceutical industry and namely to making products for the care of the oral cavity. The proposed method involves mixing the active substance with auxiliary components. In this case, enzymatic hydrolysates of coconut oil in an amount of 0.25-2.5 wt.% are used as an active ingredient. They are obtained as a result of hydrolysis of coconut oil with the help of a lipase from a pork stomach Type II with an activity of 100-500 units/mg of protein at a temperature of 37C and a pH of 7.7. Hydrolysis lasted for a period from 4 to 24 hours and was accompanied by chromatographic separation of palmitic and oleic acid residues from the hydrolyzate. Sodium fluoride, sodium lauryl sulfate, a flavouring agent and water are used as auxiliary components.

EFFECT: invention provides a new method of obtaining a bactericidal agent based on hydrolysates of coconut oil for oral care aimed at preventing caries and other diseases of the oral cavity caused by microorganisms.

5 cl, 2 tbl, 4 ex

Description

The invention relates to the cosmetic and pharmaceutical industry, namely to compositions for the care of the oral cavity, intended for the prevention of caries and other diseases of the oral cavity.

According to statistical studies [1], 99% of the world's population suffers from caries, and in many countries this figure reaches 100%. In the Russian Federation, among children aged 12 years, the prevalence of caries is from 61 to 96%. As for the adult population of Russia, the spread of the disease is 100% [2].

Despite the use of various methods of prevention and treatment, tooth decay and gingivitis are still among the most common oral diseases in children and adults. Most of the traditional preventive measures, for example, methods of improving oral hygiene, limiting the consumption of sugar-containing foods and drinks, removing dental plaque, remineralization and filling are in some cases not effective enough.

Tooth decay is an infection associated with microorganisms found in saliva. The appearance of caries is directly related to the ability of microorganisms to colonize the surface of the tooth and form biofilms or plaque on it. Biofilm formation begins with the attachment of free microorganisms to the surface. These early colonists facilitate the attachment of other bacteria by forming adhesion points. Streptococcus mutans, Lactobacilli spp. And Candida albicans are the predominant microorganisms found in plaque associated with dental caries.

The greatest role in the development of caries is played by Streptococcus mutans and Lactobacilli spp., Which exhibit greater acidogenic and acidophilic properties in comparison with other bacteria in the oral cavity. They convert food carbohydrates into acid, which lowers the pH of the mouth and dissolves the calcium phosphates of the tooth enamel, causing tooth decay. The low pH value caused by these bacteria shifts the balance of plaque microflora towards a rapid increase in Candida spp.

Currently, modern approaches are focused on the search for safe antimicrobial drugs aimed at treating the causes of oral pathologies, rather than their effects, which makes this type of therapy of great importance.

From the prior art, a composition for the care of the oral cavity is known (RF patent No. 2671835, publ. 07.11.2018), containing the following components (A), (B) and (C): (A) from 0.001% by weight to 1% by weight the mass of the compound represented by the following formula: R- (OCH ₂ CHE) _m -O- (G) _n , where R is a linear or branched alkyl group containing from 8 to 18 carbon atoms, G is a galactose residue, E is a hydrogen atom or a methyl group, m is an integer of 0 and n is an integer from 1 to 30; (B) 0.001% by mass to 0.1% by mass of one or more cationic germicidal agents selected from the group consisting of a quaternary ammonium compound and a biguanide compound; and (C) 35% by mass or more of water. The composition preferably further contains from 0.1% by weight to 1.5% by weight of a nonionic surfactant of component (D). Proposed is the use of the above composition for obtaining an agent for suppressing the accumulation of dental plaque, as well as for obtaining an agent for suppressing bad breath.

The main disadvantage of the known composition is the complex chemical composition.

An antiseptic pharmaceutical composition for oral hygiene and treatment of diseases of the oral cavity of microbial origin is known from the prior art (RF patent No. 2558061, publ. 07/27/2015), containing from 0.05 to 0.3 wt.% Hydrogen peroxide or its equivalent in the form of carbamide peroxide, from 0.001 to 0.03 wt.% eugenol, from 0.001 to 0.01 wt.% camphor, from 0.001 to 0.5 wt.% salt of zinc or other heavy metals such as silver, mercury, copper, tin, or a mixture thereof; 1 to 1.2 wt% sodium fluoride; from 2 to 7 wt.% xylitol, from 0.002 to 0.05 wt.% cetylpyridinium chloride; and excipients in an amount sufficient to bring the composition to 100%. The composition has a broad spectrum of action.

The known antiseptic pharmaceutical composition is characterized by a disadvantage - the use of heavy metal salts in the composition.

Among the oral care products, a special group is occupied by preparations containing essential oils as active ingredients, for example, Listerine®, a widely used mouthwash containing thymol and other essential oils such as menthol and eucalyptus. Other mouthwashes use triclosan or cetylpyridinium chloride as active substances and large amounts of alcohol, 25 to 30 vol.%. The alcohol is used as a carrier and as a solvent for active substances and as excipients for homogenizing a solution.

Compositions for the care of the oral cavity are known from the prior art (RF patent No. 2580642, publ. 04/10/2016), comprising an active compound from magnolia extract or a derivative thereof and an electrolyte, which is not a surfactant, in an amount effective to stabilize the composition ...

The main disadvantage of these compositions is their low antimicrobial activity.

Described is a preparation in the form of a mouthwash (US patent 5104644) containing from 0.5 to 3% w / v. hydrogen peroxide, at least 0.02% wt./about. zinc chloride, at least 0.04% wt./about. sodium lauryl sulfate, at least 0.08% wt./about. sodium citrate and from 2 to 3.5%) w / v. ethanol. Peppermint oil and menthol are used as flavoring agents. This patent claims the ability to kill bacteria, preventing dental disease.

The drug is characterized by a disadvantage - hydrogen peroxide, which is part of its composition, has a destructive effect on tooth enamel.

A known product that freshens breath, disinfects the oral cavity and contains an extract of magnolia bark and a surfactant (RF patent No. 2388457, publ. 05/10/2010), containing a delivery device into the oral cavity and an antimicrobial agent comprising magnolia bark extract and anionic surface active substance.

The main disadvantage of the product is its low antimicrobial activity.

In the literature there is information about the possibility of preventing the development of dental caries with the use of coconut oil [3]. The dominant component of coconut oil is medium-length fatty acids, which distinguishes it from other edible vegetable oils, consisting almost entirely of long-chain fatty acids. Coconut oil contains 92% saturated fatty acids, about 50% of which is lauric acid.

Although the exact mechanism of the antimicrobial action of coconut oil is still unknown, it has been hypothesized that monolauric acid and other medium-chain monoglycerides have the ability to disrupt bacterial cell walls, penetrate and destroy cell membranes, and inhibit metabolic enzymes, causing bacterial death [4]. Thus, enzymatic hydrolysates of coconut oil seem to be a promising object for the creation of new effective agents for the prevention and treatment of infectious diseases of the oral cavity, in particular caries.

During the patent search, no technical solution was found that was adopted as the closest analogue.

The technical objective of the present invention is to expand the range of bactericidal agents for the care of the oral cavity, effective against microorganisms that cause caries and other diseases of the oral cavity.

The technical result achieved by the implementation of the claimed invention consists in the development of a new method for producing a bactericidal agent based on hydrolysates of coconut oil for oral care, intended for the prevention of caries and other diseases of the oral cavity caused by microorganisms.

To achieve the specified technical result, it is proposed to use the developed method. According to the developed method of obtaining a bactericidal agent for oral care, enzymatic hydrolysis of coconut oil with lipase from a pork stomach Type II with an activity of 100-500 units / mg protein at a temperature of 37 ° C and pH 7.7 is carried out for 4-24 hours, chromatographic separation from the obtained hydrolyzate of the residues of palmitic and oleic acids, after which the obtained hydrolysates of coconut oil are mixed with auxiliary components: sodium fluoride, sodium lauryl sulfate, flavoring agent, water.

According to the invention, an oral bactericidal agent is prepared as follows. Enzymatic hydrolysis of coconut oil with lipase from a pork stomach Type II with an activity of 100-500 units / mg protein at a temperature of 37 ° C, pH 7.7 for 4-24 hours, chromatographic separation from the resulting hydrolyzate of palmitic and oleic acid residues (with using a gas chromatography-mass spectrometer), as a result of which the mass fractions of capric, lauric, stearic and myristic acids increase in the resulting product. After that, the obtained hydrolysates of coconut oil are mixed with auxiliary components in the following ratio of components, wt%:

enzymatic hydrolyzate of coconut oil - 0.25-2.5

sodium fluoride - 0.25

sodium lauryl sulfate - 2.5

flavoring additive - 1.5

water is the rest.

The invention is illustrated by the following examples.

Example 1

Coconut oil is subjected to enzymatic hydrolysis with lipase from a pork stomach Type II with an activity of 100-500 units / mg protein at a temperature of 37 ° C, pH 7.7 for 4 hours. For the obtained hydrolyzate, the fatty acid composition is analyzed by gas chromatography-mass spectrometry with using a GCMS-QP2010 Ultra / GCMS-QP2010 SE gas chromatograph-mass spectrometer (Shimadzu, Japan), the results of determining the fatty acid composition of a 4-hour hydrolyzate of coconut oil are presented in Table 1. Next, the residues of palmitic and oleic acids are chromatographically separated from the obtained hydrolyzate, in As a result, the resulting product increases the mass fraction of capric, lauric, stearic and myristic acids. Then, hydrolyzed coconut oil is mixed with auxiliary components in the following ratio of components, wt%:

4 Hrs Coconut Oil Enzymatic Hydrolyzate - 2.5

sodium fluoride - 0.25

sodium lauryl sulfate -2.5

flavoring additive - 1.5

water is the rest.

The results of assessing the antagonistic activity of the agent obtained according to example 1 in relation to representatives of the pathogenic and opportunistic microflora of the oral cavity - Streptococcus mutans and Candida albicans - are presented in table 2.

Example 2

Coconut oil is subjected to enzymatic hydrolysis with lipase from a pork stomach Type II with an activity of 100-500 units / mg protein at a temperature of 37 ° C, pH 7.7 for 8 hours. For the obtained hydrolyzate, the fatty acid composition is analyzed by gas chromatography-mass spectrometry with using a GCMS-QP2010 Ultra / GCMS-QP2010 SE gas chromatograph-mass spectrometer (Shimadzu, Japan), the results of determining the fatty acid composition of an 8-hour hydrolyzate of coconut oil are presented in Table 1. Next, the residues of palmitic and oleic acids are chromatographically separated from the obtained hydrolyzate, in As a result, the resulting product increases the mass fraction of capric, lauric, stearic and myristic acids. Then, hydrolyzed coconut oil is mixed with auxiliary components in the following ratio of components, wt%:

8 Hours Coconut Oil Enzymatic Hydrolyzate - 1.5

sodium fluoride - 0.25

sodium lauryl sulfate - 2.5

flavoring additive - 1.5

water is the rest.

The results of assessing the antagonistic activity of the agent obtained according to example 2 in relation to representatives of the pathogenic and opportunistic microflora of the oral cavity - Streptococcus mutans and Candida albicans - are presented in table 2.

Example 3

Coconut oil is subjected to enzymatic hydrolysis with lipase from a pork stomach Type II with an activity of 100-500 units / mg protein at a temperature of 37 ° C, pH 7.7 for 16 hours. For the obtained hydrolyzate, the fatty acid composition is analyzed by gas chromatography-mass spectrometry with using a GCMS-QP2010 Ultra / GCMS-QP2010 SE gas chromatograph-mass spectrometer (Shimadzu, Japan), the results of determining the fatty acid composition of a 16-hour hydrolyzate of coconut oil are presented in Table 1. Next, the residues of palmitic and oleic acids are chromatographically separated from the obtained hydrolyzate, in As a result, the resulting product increases the mass fraction of capric, lauric, stearic and myristic acids. Then, hydrolyzed coconut oil is mixed with auxiliary components in the following ratio of components, wt%:

16 Hrs Coconut Oil Enzymatic Hydrolyzate - 0.75

sodium fluoride - 0.25

sodium lauryl sulfate - 2.5

flavoring additive - 1.5

water is the rest.

The results of evaluating the antagonistic activity of the agent obtained according to example 3 in relation to representatives of the pathogenic and opportunistic microflora of the oral cavity - Streptococcus mutans and Candida albicans - are presented in table 2.

Example 4

Coconut oil is subjected to enzymatic hydrolysis with lipase from a pork stomach Type II with an activity of 100-500 units / mg protein at a temperature of 37 ° C, pH 7.7 for 24 hours. For the obtained hydrolyzate, the fatty acid composition is analyzed by gas chromatography-mass spectrometry with using a gas chromatography-mass spectrometer GCMS-QP2010 Ultra / GCMS-QP2010 SE (Shimadzu, Japan), the results of determining the fatty acid composition of a 24-hour hydrolyzate of coconut oil are presented in Table 1. Next, the residues of palmitic and oleic acids are chromatographically separated from the obtained hydrolyzate, in As a result, the resulting product increases the mass fraction of capric, lauric, stearic and myristic acids. Then, hydrolyzed coconut oil is mixed with auxiliary components in the following ratio of components, wt%:

24H Coconut Oil Enzymatic Hydrolyzate - 0.25

sodium fluoride - 0.25

sodium lauryl sulfate - 2.5

flavoring additive -1.5

water is the rest.

The results of evaluating the antagonistic activity of the agent obtained according to example 4 in relation to representatives of the pathogenic and opportunistic microflora of the oral cavity - Streptococcus mutans and Candida albicans - are presented in table 2.

From table 2 it follows that the bactericidal agents obtained according to examples 1-4 are characterized by high antagonistic activity in relation to microorganisms that play a decisive role in the development of caries (Streptococcus mutans and Candida albicans), therefore they can be used to prevent caries and other oral diseases. cavity.

Thus, the technical result of the claimed method is the development of a new method of obtaining a bactericidal agent based on hydrolysates of coconut oil for oral care, intended for the prevention of caries and other diseases of the oral cavity caused by microorganisms.

Sources of information

1.http: //rodrom.m/profilaktika-detskogo-kariesa-statistika-neumolima-v-mire-kariesom-stradaet-99.html.

2.http: //topdent.ru/articles/rasprostranennost-i-intensivnost-kariesa.html.

3. Haron, U.A. The comparative antimicrobial effect of activated virgin coconut oil (AVCO) and virgin coconut oil (VCO) against dental caries-related pathogens / U.A. Haron, Z. Abllah, N.A.M.M. Nasir // Advances in Health Science Research. 2017. Vol. 8.P. 312-317.

4. Ogbolu, D.O. In vitro antimicrobial properties of coconut oil on Candida species in Ibadan, Nigeria / D.O. Ogbolu, A.A. Oni, O.A. Daini, A.P. Oloko // J Med Food. 2007. Vol. 10 (2). P. 384-387.

Claims (25)

1. A method of obtaining a bactericidal agent for oral care, characterized in that the active substance is mixed with auxiliary components, enzymatic hydrolysates of coconut oil are used as the active substance in an amount of 0.25-2.5 wt.%, Obtained as a result of hydrolysis of coconut oils with lipase from pork stomach Type II with an activity of 100-500 U / mg protein at a temperature of 37 ° C and pH 7.7 for 4-24 hours and chromatographic separation from the hydrolyzate of palmitic and oleic acid residues , and as auxiliary components sodium fluoride, sodium lauryl sulfate, flavor, and water. 2. The method according to claim 1, characterized in that the enzymatic hydrolysis of coconut oil is carried out for 4 hours, after which the hydrolyzate is mixed with auxiliary components at the following ratio of components, wt%: 4 Hrs Coconut Oil Enzymatic Hydrolyzate - 2.5 sodium fluoride - 0.25 sodium lauryl sulfate - 2.5 flavoring additive - 1.5 water is the rest. 3. The method according to claim 1, characterized in that the enzymatic hydrolysis of coconut oil is carried out for 8 hours, after which the hydrolyzate is mixed with auxiliary components at the following ratio of components, wt%: 8 Hours Coconut Oil Enzymatic Hydrolyzate - 1.5 sodium fluoride - 0.25 sodium lauryl sulfate - 2.5 flavoring additive - 1.5 water is the rest. 4. The method according to claim 1, characterized in that the enzymatic hydrolysis of coconut oil is carried out for 16 hours, after which the hydrolyzate is mixed with auxiliary components at the following ratio of components, wt%: 16 Hrs Coconut Oil Enzymatic Hydrolyzate - 0.75 sodium fluoride - 0.25 sodium lauryl sulfate - 2.5 flavoring additive - 1.5 water is the rest. 5. The method according to claim 1, characterized in that the enzymatic hydrolysis of coconut oil is carried out for 24 hours, after which the hydrolyzate is mixed with auxiliary components at the following ratio of components, wt%: 24H Coconut Oil Enzymatic Hydrolyzate - 0.25 sodium fluoride - 0.25 sodium lauryl sulfate - 2.5 flavoring additive -1.5 water is the rest.