RU2748338C2 - Complex for prevention of oral cavity oxidative stress - Google Patents

Abstract

FIELD: dentistry.

SUBSTANCE: group of inventions relates to a complex of active components for use in dental and/or oral care products, consisting of CO₂ extract of rose hip flowers Rosa Canina, a dry extract of magnolia bark Magnolia Officinalis and an essential oil of clementine peel Citrus clementina, taken in a ratio (0.05-0.5):(0.001-0.1):(0.005-0.3) by weight, dental and/or oral care products based on it and their use. The proposed product contains the above-mentioned complex of active components in the amount of 0.1-0.3 wt%. The product may additionally contain: an enzyme (bromelain, papain, or a mixture thereof); one or more essential oil of grapefruit Citrus Paradisi, essential oil of mint Mentha Piperita, essential oil of anise Illicium Verum, essential oil of basil Ocimum Basilicum, essential oil of clove Eugenia Caryophyllus; bacteria lysate (Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus bulgaricus lysates or a mixture thereof); one or more of the water-glycerin extract of the plantain Plantago Lanceolata leaves, water-glycerin extract of clover Trifolium Pratense flowers, water-glycerin extract of Japanese Sophora Sophora Japonica buds or a mixture thereof: aqueous extract of hibiscus Hibiscus Sabdariffa flowers; dry extract of badan Bergenia Crassifolia root; fermented Punica Granatum pomegranate fruit extract. The product can be a toothpaste or mouthwash. It is proposed to use the above-mentioned complex of active components or a product based on that for the prevention of oxidative stress of the oral cavity, to reduce periodontal inflammation in a subject with such inflammation, as well as to reduce gum bleeding in a subject subject to this.

EFFECT: components of the complex have antioxidant and antibacterial effects, enhancing each other’s action to reduce periodontal inflammation.

15 cl, 8 tbl, 5 ex

Description

Technology area

The invention relates to an innovative complex of active ingredients for use in products for the care of teeth and / or oral cavity based on CO ₂ -extract of rosehip flowers Rosa Canina, dry extract of Magnolia Officinalis magnolia bark and essential oil of Citrus clementina peel, the prophylactic use of which can reduce oxidative stress of the oral cavity in a subject in need thereof.

Prior art

According to experts, oral health is one of the fundamental foundations of general human health. Diseases of the oral cavity significantly affect the quality of life of people, social aspects and cause the development of other serious diseases. Thus, periodontal disease increases the risk of developing cardiovascular diseases by 19% (Nazir Muhammad Ashraf. Prevalence of periodontal disease, its association with systemic diseases and prevention. Int J Health Sci (Qassim) 2017; 11 (2): 72-80) ...

Diseases of the oral cavity are the most common diseases that occur in people throughout their lives, causing pain and discomfort. According to the WHO, there are common oral diseases, which include tooth decay, gum disease, and trauma during dental procedures and operations. According to statistics, periodontitis is on the 6th place among the most common diseases of mankind in developed and developing countries, more than 743 million people around the world have this diagnosis. According to experts, in the United States, the state loses about $ 54 billion annually to the budget, while the cost of treating periodontal and oral cavity diseases as a whole is about $ 442 billion. The incidence in the period from 1990 to 2010 increased by 57. 3% percent. With the continued growth of urbanization, oral diseases will become even more widespread due to the discrepancy between the required amount of fluoride and its amount supplied to the body; poor quality oral hygiene; growing consumption of easily digestible sugars and transgenic fats; excessive consumption of alcohol and smoking; diabetes mellitus (Tonetti MS, Jepsen S, Jin L, Otomo-Corgel J. Impact of the global burden of periodontal diseases on health, nutrition and wellbeing of mankind: a call for global action. J Clin Periodontal. 2017; 44 (5) : 456-462).

Disturbances in the normal functioning of the natural processes of the oral cavity are caused by a wide range of factors - biological, socio-economic and psychological. Despite the fact that diseases of the oral cavity occur in people of all ages, the categories of citizens with a low socio-economic status (low income, education) are most susceptible to these diseases. WHO proposed to estimate the incidence of periodontal disease using an integral indicator - the index of need for treatment of periodontal disease (CPITN). This index ranges from 0 to 4 and shows the severity of inflammation in humans at the level of the whole population. Compared to developed countries (France, Germany, Turkey), in developing countries (India, Brazil, Iran, Saudi Arabia, Chile) among adolescents aged 15-19 years, the prevalence of bleeding is 35% higher. Tartar is similarly common: in developing countries, among adults aged 35-44 years, the prevalence is 16% higher than in developed countries. However, developed countries have the largest number of people with shallow periodontal pockets ranging in size from 4 to 5 mm. Elderly people from 65 to 74 years more often have deeper periodontal pockets measuring 6 mm compared to adults, regardless of the economic development of the countries. Thus, periodontal disease is present in 20-50% of people in different countries around the world ((Nazir Muhammad Ashraf. Prevalence of periodontal disease, its association with systemic diseases and prevention. Int J Health Sci (Qassim) 2017; 11 (2) : 72-80).

The problem of treating inflammatory periodontal diseases is currently one of the most urgent in dentistry. Gum disease is the second most common disease in humans after tooth decay, affecting the tissues around and at the base of the teeth. According to statistics from the World Health Organization (WHO), about 80% of people sooner or later face gum disease (Oral health. Fact sheets. 24 September 2018. <URL: https://www.who.int/news-room/fact- sheets / detail / oral-health>). The prevalence of periodontal disease in persons over 40 years of age exceeds 95%, and the diseases are often aggressive and have a progressive course (Omarov IA, Bolevich SB, Savateeva-Lyubimova TN, Silina EV, Sivak KV Oxidative stress and combined antioxidant energy correction in the treatment of periodontitis . J Stomatology 1, 2011, pp. 10-17).

, супероксидный анион-радикал

, гидроксидный радикал

, пероксид кислорода Н₂О₂, гидропероксидный радикал

, пероксинитрит

. АФК разрушают клетки бактерий, однако из-за неспецифичности действия также повреждают здоровые клетки: эпидермальные клетки слизистой, фибробласты соединительной ткани, эндотелий сосудов, что приводит к разрушению тканей десны, появлению кровоточивости десен, резкой болью и повышенной чувствительностью на холод. Продукты разрушения бактерий попадают в местный кровоток, активируя систему комплемента. В данный процесс также вовлечены моноциты крови, Т-лимфоциты лимфатической системы, тучные клетки тканей десен, которые в ответ на воспаление выделяют в кровь метаболиты арахидоновой кислоты, фактор активации тромбоцитов, цитокины (TNFα, IL-1β, IL-6), гистамин. Данные вещества вызывают увеличение сосудистой проницаемости и кровоточивости десен, повышение температуры за счет влияния на центр терморегуляции в гипоталамусе, способствуют дальнейшему хемотаксису нейтрофилов в место воспаления. Однако такая защитная реакция организма может переходить в следующую стадию - хроническое воспаление, вызывая прогрессирующее образование пародонтальных каналов большой глубины, разрушение корней зубов и их выпадение (Robbins and Cotran Pathologic Basis of Diseases, 8^th edition, 2009).The occurrence of inflammation is associated with insufficiently effective oral hygiene and untimely visits to the dentist for periodic preventive examinations and treatment. Inflammatory processes develop in the presence of foci of infection and pathogenic microorganisms on the oral mucosa. Insufficient care of teeth and gums, smoking, tartar, food debris between the teeth are the causes of soft tissue inflammation, swelling and redness of the gums. In total, over 800 species of bacteria have been found in the oral cavity, some of which are pathogenic microflora. With improper cleaning of teeth, a pathological increase in the depth of the periodontal sulcus occurs - a periodontal pocket, into which pathogenic microorganisms of the oral cavity penetrate and subsequently begin to actively multiply. Bacteria, using the enzyme glycosyltransferase, form biofilms from the polysaccharide dextran, protecting themselves from immunity factors: lysozyme, lactoferrin, secretory IgA, peroxidase. When foci of inflammation appear in the oral cavity, the local activity of leukocytes and T-lymphocytes of saliva can increase significantly, thus realizing a protective effect directed directly against the pathogen. It is known that phagocytes and the complement system are involved in defense mechanisms in diseases such as pulpitis, periodontitis. At the first stage, phagocytes migrate from the vascular bed to the area of inflammation through the interaction between adhesion molecules - the process of chemotaxis. When leukocytes (neutrophils and monocytes) have already entered the site of infection, they recognize the components of damaging bacteria using Toll-like receptors type 4, phagocytic, G-linked, mannose and scavenger receptors. These components can be lipopolysaccharides of the bacterial cell wall, bacterial proteoglycans and lipids, short peptides with N-formylmethionyl. Then phagocytes begin to actively absorb bacteria (process of phagocytosis) and release protective factors. Phagocytes use the enzymes NADPH oxidase and inducible NO synthase to form reactive oxygen species (ROS): hypochlorite

, superoxide radical anion

, hydroxide radical

, oxygen peroxide Н ₂ О ₂ , hydroperoxide radical

, peroxynitrite

... ROS destroy bacterial cells, however, due to the nonspecificity of action, they also damage healthy cells: epidermal cells of the mucous membrane, fibroblasts of connective tissue, vascular endothelium, which leads to the destruction of gum tissue, bleeding of the gums, severe pain and increased sensitivity to cold. Decomposition products of bacteria enter the local bloodstream, activating the complement system. This process also involves blood monocytes, T-lymphocytes of the lymphatic system, mast cells of gum tissue, which in response to inflammation release into the blood metabolites of arachidonic acid, platelet activation factor, cytokines (TNFα, IL-1β, IL-6), histamine. These substances cause an increase in vascular permeability and bleeding of the gums, an increase in temperature due to the effect on the center of thermoregulation in the hypothalamus, contribute to further chemotaxis of neutrophils to the site of inflammation. However, such a protective reaction may proceed to the next stage - chronic inflammation, causing a progressive formation of a large depth of periodontal channels, tooth decay and loss of roots (Robbins and Cotran Pathologic Basis of Diseases , 8 ^th edition, 2009).

The most common types of gum disease are gingivitis and periodontitis. The term "gingivitis" refers to inflammation of the gums without compromising the integrity of the periodontal junction. Gingivitis most often occurs due to the accumulation of microbial plaque on the teeth, as a result of non-compliance with oral hygiene procedures.

One of the factors in the onset of gingivitis is excessive plaque deposition as a result of poor oral hygiene. Dental plaque is a complex of pathogenic microorganisms of the oral cavity and their metabolic products. The causative agents of gingivitis are the following bacteria: Streptococcus oralis, Bacteroides gingivalis, Porphyromonas gingivalis, Actinomycetes comitans, Prevotella intermedia, Actinomyces israelii, rarely fungi and viruses. Risk factors for gingivitis are smoking, tartar, immunosuppressive conditions, malocclusion, diabetes mellitus, ascorbic acid deficiency in food, and others. The initial treatment for gingivitis is to reduce the number of pathogens and remove plaque from the surface of the teeth. Chronic gingivitis is associated with tartar deposition, which patients are unable to get rid of on their own. The use of antibacterial agents can reduce bacterial plaque to prevent and treat gingivitis in some patients. The American Dental Association has developed criteria for effective antibacterial agents in the treatment of gingivitis: the substance should reduce plaque and reduce the severity of inflammation for a period of at least 6 months, should be safe for humans and not cause significant side effects. Thymol, menthol, eucalyptol, methyl salicylate, chlorhexidine digluconate, and triclosan are approved for the treatment of gingivitis. However, according to the results of clinical studies, these funds are useful and effective only in case of inflammation above the gum edge, but not below (American Academy of Periodontology - Research, Science, and Therapy Committee. Treatment of plaque-induced gingivitis, chronic periodontitis, and other clinical conditions J Periodontology 2001; 72 (12): 1790-1800). Gingivitis is a reversible condition that can be eliminated with an adequate hygiene regimen. If untreated, gingivitis can progress to a destructive form of periodontal disease - periodontitis.

The term "periodontitis" refers to an inflammatory disease of the periodontal tissues, characterized by progressive destruction of the normal structure of the alveolar process of the jaw. In severe periodontitis, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Treponema denticola are most often found. Another reason is traumatic, which, in addition to the anatomical features and the presence of orthodontic structures, includes improper use of a toothbrush. Pathology is characterized by inflammation and moderate soreness of the gums, swelling of soft tissues, swelling, redness of the mucous membrane. The pain can be of different localization and occur under or around the tooth. To prevent periodontitis, an adequate hygiene regime must be observed (American Academy of Periodontology - Research, Science, and Therapy Committee. Treatment of plaque-induced gingivitis, chronic periodontitis, and other clinical conditions. J Periodontology 2001; 72 (12): 1790-1800) ...

Maintaining oral hygiene by regularly brushing your teeth will help control the development of the disease. Since gingivitis is an inflammatory gum disease, anti-inflammatory drugs such as flurbiprofen, naproxen, and ibuprofen can be used, but they only reduce the severity of inflammation and do not eliminate the original cause of the disease, inflammation from pathogenic bacteria. The best solution would be to use substances that have both antibacterial and anti-inflammatory effects (Sekino S, Ramberg P, Lindhe J. The effect of systemic administration of ibuprofen in the experimental gingivitis model. J Clin Periodontal. 2005; 32 (2): 182-187 ; Rosin M, Kahler ST, Hessler M, Schwahn Ch, Kuhr A, Kocher T. The effect of a dexibuprofen mouth rinse on experimental gingivitis in humans. J Clin Periodontal. 2005; 32 (6): 617-621).

The state of the periodontium in normal conditions and in periodontitis largely depends on the functioning of the cells of innate immunity - in particular, neutrophils. The destruction of bacteria by neutrophils normally occurs without the development of an active phase of inflammation. Smoking, poor diet, physical damage leads to an increase in the colonization of the microbial community by the bacterium P. gingivalis, as a result, neutrophils enter the connective tissue, but do not migrate to the site of infection, which leads to increased inflammation. As a result, there is an additional attraction of neutrophils to tissues and their destruction (Uriarte SM, Edmisson JS, Jimenez-Florez E. Human neutrophils and oral microbiota: a constant tug-of-war between a harmonious and a discordant coexistence. Immunological Reviews 2016; 273: 282-298; Olsen I, Lambris JD, Hajishengallisc G. Porphyromonas gingivalis disturbs host-commensal homeostasis by changing complement function. J Oral Microbiol. 2017; 9 (1): 1340085).

The immune response in inflammation involves not only neutrophils, but also T-lymphocytes. Immunity plays a key role in the development of the pathogenesis of gingivitis and periodontitis. Studies show that regulatory T-lymphocytes (T-regulatory) suppress the development of the immune response by suppressing the activity of other immune cells and their population is greatly increased in periodontitis and severe gingivitis. An association was found between the size of the population of regulatory T-lymphocytes (CD4 +, CD25 +, FoxP3 +) and the activity of the transcription factor NF-κB (receptor activator of nuclear factor kappa-B ligand (RANKL) pathway in periodontitis and gingivitis (Arul D, Rao S. Isolation of naturally induced T-regulatory cells in gingival tissues of healthy human subjects and subjects with gingivitis and chronic periodontitis. Cureus 2019; 11 (3): e4283).

In the course of studies on the morphological comparison of gum tissue, it was found that in chronic gingivitis and periodontitis, the extracellular matrix was severely damaged, and the fibers of type I collagen were reorganized. This process also involved a large number of inflammatory cells (monocytes, macrophages, lymphocytes) and permanent cells (fibroblasts, epithelial cells and endothelial cells), which secreted cytokines and protease enzymes that destroy the extracellular matrix and disrupt the folding of collagen fibers. It should be noted that the pathogenic microorganisms themselves isolated matrix metalloproteases involved in the reorganization of the extracellular matrix of the gingival connective tissue (Almeida T, Valverde T, Martins-Junior P, Ribeiro N, Kitten G, Carvalhaes L. Morphological and quantitative study of collagen fibers in healthy and diseased human gingival tissues.Rom J Morphol Embryol 2015; 56 (1): 33-40). In periodontitis, CD68 + mononuclear cells: monocytes, macrophages - secrete a large amount of type 12 matrix metalloprotease (macrophage metalloelastase) due to the influence of the CSF-2 factor and the COX-2 pathway. Matrix metalloprotease type 12 destroys extracellular matrix proteins - tropoelastin and fibronectin, which together with collagen make up the basis of the connective tissue of the oral gums. Also, this enzyme mediates the inflammatory response by releasing the pro-inflammatory cytokine TNFα from lymphocytes, macrophages, fibroblasts (Holsmstrom SB, Clark R, Zwicker S, Bureik D, Kvedaraite E, Bernasconi E, Thu Nguyen Hoang A, Johannsen G, Marsland EA , Svensson M. Gingival tissue inflammation promotes increased matrix metalloproteinase-12 production by CD200R ^low monocyte-derived cells in periodontitis. The Journal of Immunology 2017; 199: 1-14).

Substances that take part in inflammatory diseases are being studied, but there is evidence of certain changes in the vital activity of cells. Thus, data were obtained indicating an increased amount of expression of the beta-catenin protein and the secreted transmembrane protein frizzled-1, depending on the size of the periodontal damage. Normally, these proteins regulate cell proliferation and tissue regeneration through the Wnt signaling pathway, however, with an increased level of expression, they can provoke the development of periodontitis (Chi Y, Xu C, Sun X, Li D, Wang H, Wang M, He X. Expression and significance of secreted frizzled-related protein 1 and β-catenin in gingival tissue of patients with chronic periodontitis. West China Journal of Stomatology 2018; 36 (3): 257-261).

With inflammatory diseases of the oral cavity, the state of not only the gums and teeth changes, but also the qualitative and quantitative composition of saliva. In the course of studies, it was found that with gingivitis and periodontitis, the amount of sialic acid in saliva increases two and three times, respectively, compared with the amount of the substance in the absence of inflammatory diseases of the oral cavity. Sialic acid is an important biomarker, the amount of which correlates with the severity of systemic inflammation. This compound is part of the glycolipids and glycoproteins, which are in a mobile state in the cell membrane. The function of sialic acid is expressed in the regulation of innate immunity. The most well-known form of sialic acid is N-acetylneuraminic acid. Sialic acid increases the synthesis of protective molecules during oxidative stress in periodontitis and gingivitis (Hernandez-Cedillo A, Garcia Valdivieso MG, Hernandez-Arteaga AC, Patino Marin N, Hernandez AAV, Jose-Yacaman M, Navarro-Contreras HR. Determination of sialic acid levels by using surface-enhanced Raman spectroscopy in periodontitis and gingivitis. Oral Dis. 2019). The enzyme neuraminidase of pathogenic microorganisms has been suggested to break down power acid, promoting plaque formation and further progression of periodontitis (Ide, M., McPartlin, D., Coward, PY, Crook, M., Lumb, P., & Wilson, RF Effect of treatment of chronic periodontitis on levels of serum markers of acute-phase inflammatory and vascular responses.Journal of Clinical Periodontology 2003; 30 (4): 334-340; Jawzali, JI Association between salivary sialic acid and periodontal health status among smokers. The Saudi Dental Journal 2016; 28 (3): 124-135; Jwan I. Jawzaly, HGH & B. MA Relationships of salivary sialic acid and its fraction in periodontitis with demographic properties and medical history. International Journal of Dental Research & Development (IJDRD) 2013 ; 3 (2): 17-26; Rathod, SR, Khan, F., Kolte, AP, & Gupta, M. Estimation of salivary and serum total sialic acid levels in periodontal health and disease.Journal of Clinical and Diagnostic Research 2 014; 8 (9): ZC19-ZC21). According to the results of studies by scientists from South Korea, with periodontitis, the amount of IL-8 and type 9 matrix metalloprotease increases in saliva, therefore, they can be used as biomarkers of pronounced inflammation of the oral cavity along with sialic acid. IL-8 is a highly active pro-inflammatory cytokine that is secreted from macrophages and epithelial cells to attract neutrophils to the site of inflammation. Type 9 matrix metalloprotease is the main enzyme that breaks down collagen in the development of gingivitis and periodontitis (Kim HD, Shin MS, Kim HT, Kim MS, Ahn YB. Incipient periodontitis and salivary molecules among Korean adults: association and screening ability. J Clin Periodontal. 2016 ; 43 (12): 1032-1040).

, гидроксидный радикал

, пероксид кислорода Н₂О₂, гидропероксидный радикал

. Избыток АФК способствует свободно-радикальному перекисному окислению липидов (ПОЛ), что повреждает ткани, прежде всего биологические мембраны клеток. При появлении большого количества свободных радикалов нарушается транспорт электронов в дыхательной цепи митохондрий; разобщение окислительного фосфорилирования под действием ПОЛ ведет к глубокому дефициту энергии; изменяются функции ферментов, углеводов и белков, в том числе белков ДНК и РНК. В результате клеткой утрачиваются регуляторные функции, появляются аномальные белки и стимулируются, помимо прямого повреждающего действия, вторичные деструктивные процессы. Глубокие нарушения мембранного и, в последующем, тотального устройства клетки приводит к ее гибели. Данный процесс носит название оксидативного стресса (Omarov I.A., Bolevich S.B., Savateeva-Lyubimova T.N., Silina E.V., Sivak K.V. Oxidative stress and combined antioxidant energy correction in the treatment of periodontitis. J Stomatology 1, 2011, p. 10-17).Another factor in the onset of gingivitis is oxidative stress (oxidative stress). In recent decades, the problems of free radical processes (FRP) have been widely studied, the role of which has been proven in a variety of nosologies, however, the number of studies devoted to the state of oxidative stress in patients with periodontal disease is not numerous. PSA is a general biological mechanism of tissue protection and damage. Normally, they participate in energy processes, in the transport of electrons in the respiratory mitochondrial chain, cell proliferation and differentiation, in the regulation of enzyme activity, etc. In addition, CRP is a necessary link in any inflammation associated with the production of reactive oxygen species (ROS) by phagocytes. This evolutionarily developed secretory function of phagocytes is necessary for the destruction of pathogenic bacteria, however, a sharp increase in oxygen consumption during phagocytosis leads to the fact that, instead of reducing O ₂ to H ₂ O, leukocytes mainly generate the following ROS: superoxide anion radical

, hydroxide radical

, oxygen peroxide Н ₂ О ₂ , hydroperoxide radical

... An excess of ROS promotes free radical lipid peroxidation (LPO), which damages tissues, primarily the biological membranes of cells. When a large number of free radicals appear, the transport of electrons in the respiratory chain of mitochondria is disrupted; uncoupling of oxidative phosphorylation by LPO leads to a deep energy deficit; the functions of enzymes, carbohydrates and proteins, including DNA and RNA proteins, change. As a result, the cell loses its regulatory functions, abnormal proteins appear and, in addition to the direct damaging effect, secondary destructive processes are stimulated. Deep violations of the membrane and, subsequently, the total structure of the cell leads to its death. This process is called oxidative stress (Omarov IA, Bolevich SB, Savateeva-Lyubimova TN, Silina EV, Sivak KV Oxidative stress and combined antioxidant energy correction in the treatment of periodontitis. J Stomatology 1, 2011, p. 10-17).

In the course of research, it was revealed that with periodontitis in saliva, the amount of such a biomarker as malondialdehyde, which occurs during LPO and oxidative stress, increases. It has been shown that malic dialdehyde is able to react with DNA, forming DNA adducts, primarily the mutagenic M ₁ G - the product of the addition of malonic aldehyde to guanine (Cherian DA, Peter T, Narayanan A, Madhavan SS, Achammada S, Vynat GP. Malondialdehyde as a marker of oxidative stress in periodontitis patients. J Pharm Bioallied Sci. 2019; 11 (2): S297-300).

Considering that a significant role in the pathogenesis of gingivitis and periodontitis is played by the bacterial flora, which is closely related to the activity of free radicals and protective reactions of the immune system, PSA is one of the links in the development and course of gingivitis and periodontitis. The PSA imbalance in periodontal disease affects the oxygen stage of oxidative stress to a greater extent and lasts for a long time, and therefore it becomes necessary to use antioxidants and energy correctors for inflammation of the gums to improve the course of the disease (Omarov IA, Bolevich SB, Savateeva-Lyubimova TN, Silina EV, Sivak KV Oxidative stress and combined antioxidant energy correction in the treatment of periodontitis. J Stomatology 1, 2011, p. 10-17).

For the prevention and treatment of periodontal inflammation, it is advisable to use medicinal plants and their extracts due to the gentle effect on the gum tissue, the high efficiency of the entire complex of biologically active substances (BAS) and the low incidence of side effects after use in comparison with drugs, such as flower extract rosehip, citrus essential oil, hibiscus extract, clove essential oil, magnolia bark extract, etc.

Rosehip has long been known to Russian medicine and is used in various forms: rosehip fruits, fruit infusion, vitamin preparations, dry extract, rosehip syrup, the drug "Holosas", which has a choleretic effect, etc. Distinguish between low-vitamin and high-vitamin types of rosehips, which are regulated in the State Pharmacopoeia XIV editions and collection of which is allowed to obtain medicinal plant materials. So, high-vitamin species include May rosehip (Rosa majalis), needle rose (Rosa acicularis), wrinkled rosehip (Rosa rugosa), Daurian rosehip (Rosa davurica) and other representatives. In them, the content of ascorbic acid as the main BAS is in the range from 1 to 18%. Low-vitamin fruits of dog rose (Rosa canina), small-flowered wild rose (Rosa floribunda) contain less than 1% ascorbic acid.

Despite the fact that in the cosmetic industry the extract of rose hips is actively used, the authors of the present invention used the extract of flowers / rose hips. In rose hips, the main biologically active compounds are ascorbic acid, which has a pronounced antioxidant effect, and flavonoids. That is why rose hips are standardized for the content of ascorbic acid and flavonoids. It should be noted that the content and composition of flavonoids in rose hips is less represented than in rose hips, therefore the antioxidant effect of the fruit extract is moderate. Ascorbic acid is an unstable compound and is easily oxidized during extraction from rose hips, so its biological activity is reduced to a minimum. Ascorbic acid in the petals of rosehip flowers contains a minimum amount, because the main BAS are essential oil, hydrolyzable tannins and polysaccharides, flavonoids and anthocyanins (Nowak R, Olech M, Pecio L, Oleszek W, Los R, Malmc A, Rzymowskad J. Cytotoxic, antioxid , antimicrobial properties and chemical composition of rose petals. J Sci Food Agric 2014; 94: 560-567).

Rosehip petals contain the largest spectrum of substances with antioxidant and anti-inflammatory activity, therefore, the use of an extract from flowers is most justified in inflammatory diseases of the oral cavity, for example, gingivitis and periodontitis. Flavonoids and anthocyanins neutralize reactive oxygen species at the site of inflammation, essential oil provides wound healing and antibacterial effects, and hydrolyzed tannins form a protective film on the gum surface to protect the gums and reduce pain sensitivity. Thus, it is the rosehip flower extract that has a versatile and effective effect in inflammatory diseases of the oral cavity, in contrast to the rosehip extract.

Various solvents and methods can be used to extract biologically active substances from rosehip flowers. Traditional methods use ethyl alcohol, a mixture of glycerin and water as extractants. There is an innovative method for obtaining extracts - cryoextraction. Cryoextraction is a cold extraction method using solvents with a low boiling point and which are in a gaseous state at ambient temperatures. Cryoextracts contain more biologically active substances than conventional aqueous extracts. To provide the required therapeutic effect, the entire complex of biologically active substances from medicinal plants is required. The use of water as an extractant does not allow the extraction of all biologically active substances, therefore the effectiveness of such an extract is less. The use of organic solvents (ethyl alcohol, glycerin, etc.) with an amphiphilic nature makes it possible to obtain extracts of more valuable composition with the presence of polyphenolic compounds with antioxidant activity; therefore, they can also be used to treat inflammatory diseases of the oral cavity.

However, the most innovative is the production of extracts using carbon dioxide CO _{2, which is} compressed to a liquid state. At the same time, organic solvents are not able to ensure the extraction of the full complex of biologically active substances due to the different solubility of certain groups of biologically active substances. In CO ₂ extracts, liquefied carbon dioxide, used as a solvent (solvent), extracts essential oils, pigments, fat-soluble fraction of polyphenolic compounds, terpenes. The resulting extract possesses the entire set of biologically active substances, since the extraction is carried out at higher pressures. Due to the lipophilic nature of CO ₂ -extracts are in many respects related to some components of the hydrolipid structures of the skin, therefore, biologically active substances in the composition of such extracts more easily penetrate into the skin. It should be noted that CO ₂ extract is more effective than aqueous extracts.

CO ₂ -extract of rosehip flowers is obtained from various plant species of the genus Rosa of the Rosaceae family, which also differ in biochemical composition depending on the place and growing conditions. Among the list of biologically active substances, the most active ingredients of rosehip are phenolic acids (caffeic acid, salicylic, coumaric acids), flavonoids (rutin, isoquercetin, tilyroside, quercetin, kaempferol-3-O-rutinoside, apigenin-7-O-glucoside), essential oils and polysaccharides consisting of monosaccharide residues: glucose, mannose, rhamnose, galactose, xylose, fructose, fucose and arabinose. Rosehip flowers also contain hydrolyzable tannins (rugozin D and tellimagrandin II), linool, phenylethyl alcohol, citronellol, α-bisobolol (Nowak R, Olech M, Pecio L, Oleszek W, Los R, Malmc A, Rzymowskad J. Cytotoxic, antioxidant antimicrobial properties and chemical composition of rose petals. J Sci Food Agric 2014; 94: 560-567).

является одним из чрезвычайно активных свободных радикалов из-за способности вызывать быстро прогрессирующее перекисное окисление липидов, мутации в ДНК и нарушение структуры белков-ферментов. Супероксидный анион-радикал

является предшественником АФК, способствуя образованию других свободных радикалов.With inflammation, the activity of cells of the immune system increases, which secrete a large amount of ROS to eliminate pathogenic microorganisms. Hydroxide radical

is one of the extremely active free radicals due to its ability to cause rapidly progressive lipid peroxidation, mutations in DNA and disruption of the structure of protein-enzymes. Superoxide radical anion

is a precursor of ROS, promoting the formation of other free radicals.

и гидроксидного радикала

. Антиоксидантная активность полисахаридов шиповника зависит от качественного состава из моносахаридов, молекулярной массы, типа гликозидной связи и содержания уроновой кислоты. Отмечено, что чем больше уроновой кислоты в составе полисахаридов, тем выше антиоксидантная активность (Zhang С, Zhao F, Li R, Wu Y, Liu S, Liang Q. Purification, characterization, antioxidant and moisturepreserving activities of polysaccharides from Rosa rugosa petals. International Journal of Biological Macromolecules, 2018).Polysaccharides reduced the severity of oxidative stress by stabilizing the superoxide anion radical

and hydroxide radical

... The antioxidant activity of rosehip polysaccharides depends on the qualitative composition of monosaccharides, molecular weight, type of glycosidic bond and uronic acid content. It is noted that the more uronic acid in the composition of polysaccharides, the higher the antioxidant activity (Zhang C, Zhao F, Li R, Wu Y, Liu S, Liang Q. Purification, characterization, antioxidant and moisture preserving activities of polysaccharides from Rosa rugosa petals. International Journal of Biological Macromolecules, 2018).

It was found that rosehip flower extract inhibited tyrosinase, which catalyzes the oxidation of phenolic compounds in cells, and had an antibacterial effect (Ren G, Xue P, Sun X, Zhao G. Determination of the volatile and polyphenol constituents and the antimicrobial, antioxidant, and tyrosinase inhibitory activities of the bioactive compounds from the by-product of Rosa rugosa Thunb.var.plena Regal tea. BMC Complement Altern Med. 2018; 18 (1): 307). Antibacterial activity was noted against 8 bacterial species: S. epidermidis, S. aureus, B. subtilis, M. luteus, E. coli, K. pneumoniae, P. aeruginosa, P. Mirabilis (Nowak R, Olech M, Pecio L, Oleszek W, Los R, Malmc A, Rzymowskad J. Cytotoxic, antioxidant, antimicrobial properties and chemical composition of rose petals. J Sci Food Agric 2014; 94: 560-567).

Currently, much attention is paid to citrus fruits, including clementine. Clementine essential oil is obtained from the peel of the Citrus clementina fruit of the Citrinae family. Clementine essential oil contains a wide range of biologically active substances - in particular, terpenoids: limonene (about 95%), myrcene (about 0.75%), linalol (about 0.5%), alpha-pinene (about 0.5%) , sabinene (about 0.5%), alpha-terpineol, terpinene, etc. However, not only terpenoids have a pronounced therapeutic effect. The high content of flavonoids in clementine is responsible for antioxidant, anti-inflammatory, anti-allergic and anti-cancer properties. Flavonoids are represented by flavones (diosmin, luteolin and sinensetin), flavonones (naringin, hesperidin, neohesperidi), flavonols (rutin, quercetin, kaempferol) and their glycosides. It is known that hesperidin has P-vitamin activity, that is, it inhibits the activity of hyaluronidase. This increases the concentration of hyaluronic acid, which increases the elasticity of the capillaries and reduces their permeability, which is useful in the progression of inflammation. In addition, together with naringin, hespyridin prevents copper-dependent oxidation of low density lipoproteins, which is important in oxidative stress (Alvarez R, Carvalho CP, Sierra J, Lara O, Cardona D, Londono-Londono J. Citrus juice extraction systems: effect on chemical composition and antioxidant activity of clementine juice. J Agric Food Chem 2012; 60 (3): 774-781). However, in the course of research it was revealed that the individual compound limonene showed a pronounced inhibitory activity of elastase and collagenase, which are involved in the destruction of the extracellular matrix. Clementine essential oil contains more than 95% limonene, therefore it has a pronounced stabilizing and anti-inflammatory effect and prevents oxidative stress (Fraternale D, Flamini G, Ascrizzi R. In vitro anticollagenase and antielasttase activities of essential oil of Helichrysum italicum subsp.italicum (Roth) G. Don. J Med Food 2019).

I.,

S.,

D.,

P.A., Del

R.,

I.,

A.,

J.F. and Segura-Carretero A. Characterization of phenolic compounds anthocyanidin, antioxidant and antimicrobial activity of 25 varieties of Mexican Roselle (Hibiscus sabdariffa). Industrial Crops Products, 2015; 69: 385-394). Механизм действия органических кислот связан со снижением концентрации ионов кальция в клетках путем образования хелатных комплексов, что способствует снижению выраженности воспаления. В оксидативном стрессе главное место занимают ионы кальция, способствующие активации ферментов и клеток воспаления, секреции цитокинов и провоспалительных веществ.Dry hibiscus extract is obtained from hibiscus flowers Hibiscus sabdariffa of the Malvaceae Malvaceae family. Hibiscus extract contains a wide range of biologically active substances: anthocyanins (mainly cyanidin 3-O-sambubioside, delphinidin 3-O-sambubioside, delphinidin, delphinidin-3-glucoside, cyanidin-3-glucoside, etc.); gossipetine, protocatechoic acid, hibisic acid, organic hydroxy acids (citric, malic, tartaric, ascorbic acids), flavonoids (quercetin-3-sambulozide and rutinoside, myricetin-3-sambuloside, hibiscetin, hibiscentrin, phenolcarbolic acid complex) coumaric acids), alpha-tocopherol, chlorogenic acid (3-O-caffeylquinic acid) and 5-O-caffeylquinic acid. A complex of 14 phenolic compounds in the experiment demonstrates a pronounced antioxidant effect (

I.,

S.,

D.,

PA, Del

R.,

I.,

A.,

JF and Segura-Carretero A. Characterization of phenolic compounds anthocyanidin, antioxidant and antimicrobial activity of 25 varieties of Mexican Roselle (Hibiscus sabdariffa). Industrial Crops Products, 2015; 69: 385-394). The mechanism of action of organic acids is associated with a decrease in the concentration of calcium ions in cells through the formation of chelate complexes, which helps to reduce the severity of inflammation. In oxidative stress, the main place is occupied by calcium ions, which promote the activation of enzymes and inflammatory cells, the secretion of cytokines and pro-inflammatory substances.

M, Oliveira M. Beatrix PP, Isabel Ferreira IC.F.R. Hibiscus sabdariffa L. as a source of nutrients, bioactive compounds and colouring agents. Food Research International 2017).The main compounds - anthocyanins perform an antioxidant function, stabilizing reactive oxygen species and forming inactive intermediates due to their reducing properties due to hydroxyl groups (Deli M, Ndjantou EB, Ngatchic Metsagang JT, Petit J, Yanou NN, Scher J. Successive grinding and sieving as a new tool to fractionate polyphenols as antioxidants of plants powders: Application to Boscia senegalensis seeds, Dichostachys glomerate fruits, and Hibiscus sabdariffa calyx powders. Food Sci Nutr. 2019; 7: 1795-1806; Ramakrishna BV, Jayaprakasha GK, Jena BS, Singh RP . Antioxidant activities of roselle (Hibiscus abdariffa) calyces and fruit extracts. J Food Sci Technol 2008; 45: 223-227). It is currently believed that delphinidin, as the main anthocyanin in hibiscus, has the highest antioxidant activity of all known anthocyanins. Delphinidin inhibits the activity of COX-2, an inducible NO synthase, TNFα and can terminate the free radical chain induced by copper ions and ascorbic acid (Medina-Carrillo, R.E., Madrigal-Santillan, E., Machuca-Sanchez, M.L. , Balois-Morales, R., Jimenez-Ruiz, EI, Valadez-Vega, C., Sumaya-Martinez, MT Free radical scavenging properties and their relationship bioactive compounds content of dehydrate calyces of roselle (Hibiscus sabdariffa L.) African Journal of Agricultural Research 2016; 10 (11): 1203-1210). Delphinidin, due to the content of the largest amount of hydroxyl, is able to form stable intermediates, intercepting electrons from free radicals and ROS. The anthocyanins of hibiscus flower extract (delphinidin, cyanidin-3-O-glycoside) have been found to inhibit the formation of malondialdehyde, a byproduct of LPO. Chlorogenic acid (trans-3-O-caffeylquinic acid) has a pronounced anti-inflammatory activity. The hibiscus flower extract exhibited not only antioxidant and anti-inflammatory functions, but also antibacterial and antifungal effects. Thus, the extract slowed down the growth of Bacillus cereus, Trichoderma viride, and Aspergillus ochraceus at minimum concentrations, while Micrococcus flavus, Staphylococcus aureus, Listeria monocytogenes, Pseudomonas aeruginosa and Salmonella typhimirium at higher concentrations (Jabeur, Bar

M, Oliveira M. Beatrix PP, Isabel Ferreira IC.FR Hibiscus sabdariffa L. as a source of nutrients, bioactive compounds and coloring agents. Food Research International 2017).

One of the pathogenic microorganisms of the oral cavity, Streptococcus mutans, produces glycosyltransferase for the formation of dextran biofilms, which protect it from the action of human immunity factors and contribute to the development of caries. However, polyphenols, flavonoids and anthocyanins inhibit the activity of the enzyme glycosyltransferase, the main enzyme of pathogenic microorganisms of the oral cavity (Ren Z, Chen L, Li J, Li Y. Inhibition of Streptococcus mutans polysaccharide synthesis by molecules targeting glycosyltransferase activity. J Oral: Microbiol. 2016; 8 31095). Delphinidin is an unstable compound and can be converted to gallic acid, which has an antioxidant effect. It was found that delphinidin and its metabolite, gallic acid, promote the differentiation of regulatory T-lymphocytes by releasing the anti-inflammatory cytokine TGF-β. This growth factor is believed to play an important role in the regulation of the immune system (Fakeye TO, Pal A, Bawankule DU, Khanuja SPS. Immunomodulatory effect of extracts of Hibiscus sabdariffa L. (Family Malvaceae) in a mouse model. Phytother Res 2008; 22 (5): 664-668). Through the Foxp3 protein, it affects regulatory T-lymphocytes and T-helpers of type 17, and also blocks the activation of T-lymphocytes and macrophages. Thus, delphinidin has immunosuppressive properties and reduces the severity of inflammation (Hyun KH, Gil KC, Kim SG, Park SY, Hwang KW. Delphinidin chloride and its hydrolytic metabolite gallic acid promote differentiation of regulatory T cells and have an anti-inflammatory effect on the allograft model J Food Sci 2019; 84 (4): 920-930).

Dry extract of magnolia bark is obtained from Magnolia officinalis Magnolia officinalis of the family Magnoliaceae. Magnolia extract is recommended for use in cosmetic products (Liu, Z., Zhang, X., Cui, W., et al. Evaluation of short-term and subchronic toxicity of magnolia bark extracts in rats. Regul. Toxicol. Pharmacol., 2007 ; 49 (3): 160-171). Magnolia bark has long been known in Japanese and Chinese medicine as a remedy for the treatment of gastrointestinal diseases, anxiety and depression, diseases of the nervous system, bronchial asthma and allergic manifestations; moderately reduces headache, muscle pain and fever. The main active ingredients in the extract are phenolic compounds: magnolol (5,5'-diallyl-2,2'-dihydroxybiphenyl) and honokiol (5,3'-diallyl-2,4'-dihydroxybiphenyl), which are accompanied by sesquiterpene alcohols - α and β-eudesmols, alkaloids, derivatives of aporphine and benzyltetrahydroisoquinoline, mineral salts: calcium, potassium, magnesium, iron salts.

Magnolol and honokiol are lignans, hydroxybiphenolic compounds in terms of chemical structure. The content of these lignans can range from 40% to 90%, depending on the type of medicinal plant used. During the study, it was found that the extract of magnolia bark has anti-cancer, antioxidant, anti-inflammatory and other effects, which confirms the high value of this plant material.

Interest in Magnolia Bark Extract emerged after the publication of the results of a study according to which Magnolia Bark Extract reduced the number of Porphyromonas gingivalis and Fusobacterium nucleaticum, and also inhibited the enzyme glycosyltransferase, which is involved in the formation of bacterial plaque and inflammation of the gums. The presence of key pathogens, P. gingivalis and F. nucleaticum, is also associated with the risk of oropharyngeal squamous cell carcinoma, pancreatic cancer, and colon cancer.

Magnolol and Honokiol suppress activation of the NF-kB pathway and expression of type 1 matrix metalloprotease, preventing inflammation and degradation of extracellular matrix proteins. NF-kB stimulates the transcription of genes of pro-inflammatory factors, promotes the release of cytokines and NO from macrophages. Magnolol and Honokiol have also been shown to inhibit the MAPK pathway involved in inflammation.

Honokiol has a pronounced antioxidant effect by stabilizing superoxide and peroxide radicals. Magnolol and honokiol inhibit the synthesis of TNFα and NO, as well as reduce the synthesis of eicosanoids by inhibiting the enzymes of the arachidonic acid cycle: phospholipase A ₂ , COX-2, 5-LOX and LTS-4. Magnolia extract is effective against gram-positive bacteria Streptococcus mutans, Staphylococcus aureus, Listeria monocytogenes, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Capnocytophaga gingivalis, Prevotella intermedia, Micrococcus luteicus, and Tungsten mushrooms It is assumed that lignans of the magnolia bark increase the permeability of bacterial cell membranes and interact with transpeptidase, an enzyme for the synthesis of the cell wall of gram-positive bacteria (Poivre M, Duez P. Biological activity and toxicity of the Chinese herb Magnolia officinalis Rehder & E. Wilson (Houpo) and its constituents . Journal of Zhejilang University-SCIENCE B. 2017; 18 (3): 194-214).

The safety of magnolol and honokiol when ingested in terms of mutagenicity has been confirmed in the course of studies, therefore, pure substances and magnolia bark extract can be used orally (Sarrica A, Kirika N, Romeo M, Salmona M, Diomede L. Safety and toxicology of magnolol and honokiol. Planta Med. 2018; 84 (16): 1151-1164).

The essential oil of clove has long been known for its action in perfumery and cosmetics. So, in the 1st millennium BC. in China, cloves were used as a flavoring and antiseptic - before the imperial audience, a clove bud was to be chewed to get rid of halitosis. Today, clove oil is used in medicine, cooking, aromatherapy, cosmetology, and perfumery.

JJ, Vargas

LY. Semisynthetic eugenol derivatives as antifungal agents against dermatophytes of the genus Trichophyton. J Med Microbiol. 2019).Clove essential oil is obtained from the immature flower buds of the clove tree Syzygium aromaticum of the Myrtaceae family Myrtaceae. The main BAS of clove essential oil is eugenol - the oil is 60-90% composed of this substance. This substance is from the class of phenols, named after the shrub of the Myrtle family Eugenia, known for its bactericidal action. In addition to eugenol, clove oil contains eugenol acetate, caryophyllene and salicylic acid methyl ester, which is responsible for anti-inflammatory properties. Eugenol has antibacterial, anti-inflammatory, antifungal and immunomodulatory properties (Pinto SML, Rivera Y, Herrera Sandoval LV, Lizarazo JC,

JJ, Vargas

LY. Semisynthetic eugenol derivatives as antifungal agents against dermatophytes of the genus Trichophyton. J Med Microbiol. 2019).

L, Tvarijonaviciute A, Tort L, Mancera JM. Transport and recovery of gilthead sea bream (Sparus aurata L.) sedated with clove oil and MS222: effects on oxidative stress status. Front Physiol. 2019; 10: 523).Clove essential oil has been shown to have an antioxidant effect by neutralizing free radicals during oxidative stress and altering the expression of enzymes that suppress oxidative stress in cells. Increased expression of genes for glutathione peroxidase 1 (gpx1), catalase (cat), zinc-dependent superoxide dismutase (sod2), glutathione reductase (gr), and glutathione S-transferase 3 (gst3) reduce oxidative stress. An increased level of heat shock proteins 70 (HSP70) indicates the severity of inflammation and the body's response to preserving the structure and properties of intracellular proteins (Teles M, Oliveira M, Jerez-Sera I,

L, Tvarijonaviciute A, Tort L, Mancera JM. Transport and recovery of gilthead sea bream (Sparus aurata L.) sedated with clove oil and MS222: effects on oxidative stress status. Front Physiol. 2019; 10: 523).

The essence of the invention

In a first aspect, the claimed invention relates to a complex of active ingredients for use in dentifrice and / or oral care products, consisting of: CO ₂ extract of Rosa Canina rose hips, dry extract of Magnolia Officinalis magnolia bark and essential oil of Citrus clementina peel, taken in the ratio (0.05-0.5) :( 0.001-0.1) :( 0.005-0.3) by weight.

In a second aspect, the claimed invention relates to a dentifrice and / or oral care product that contains a complex of active ingredients according to the invention in an amount of 0.1 to 0.3% by weight.

The dentifrice and / or oral care agent may further comprise an enzyme suitable for use in dentifrice and / or oral care products.

The dentifrice and / or oral care product may further comprise an enzyme suitable for use in dentifrice and / or oral care products, wherein the enzyme suitable for use in dentifrice and / or oral care products is bromelain, papain, or a mixture thereof.

The dentifrice and / or oral cavity can further comprise one or more of: Citrus Paradisi grapefruit essential oil, Mentha Piperita peppermint essential oil, Illicium Verum anise essential oil, Ocimum Basilicum essential oil, Eugenia Caryophyllus clove essential oil.

The dentifrice and / or oral cavity can further comprise a bacterial lysate.

The dentifrice and / or oral cavity can further comprise a bacterial lysate, wherein the bacterial lysate is a lysate of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus fermentus, Lactobacillus fermentum, Lactobacterium bifidum.

The dentifrice and / or oral cavity care product may additionally contain one or more of: water-glycerin extract of plantain leaves Plantago Lanceolata, water-glycerin extract of Trifolium Pratense clover flowers, water-glycerin extract of Japanese sophora buds Sophora Japonica, or a mixture thereof.

The dentifrice and / or oral cavity can further comprise an aqueous extract of Hibiscus Sabdariffa hibiscus flowers.

The dentifrice and / or oral cavity care product may additionally contain dry extract of Bergenia Crassifolia bergenia root.

The dentifrice and / or oral cavity can further comprise fermented Punica Granatum pomegranate fruit extract.

The dentifrice and / or oral cavity can be a toothpaste or mouthwash.

In a third aspect, the claimed invention relates to the use of an active ingredient complex according to the invention or a dentifrice and / or oral care agent according to the invention for the prevention of oxidative stress in the oral cavity.

In a fourth aspect, the claimed invention relates to the use of a complex of active ingredients according to the invention or a dentifrice and / or oral care agent according to the invention for reducing periodontal inflammation in a subject having such inflammation.

In a fourth aspect, the claimed invention relates to the use of a complex of active ingredients according to the invention or a dentifrice and / or oral care product according to the invention for reducing gingival bleeding in a subject subject.

Detailed description of the invention

An aspect of the claimed invention is a complex of active ingredients for use in dentifrice and / or oral care products. The beneficial effect of such a combination of components is due to the active substances contained in them. The complex of active ingredients according to the invention contains or consists of: CO ₂ -extract of rosehip flowers Rosa Canina, dry extract of Magnolia Officinalis magnolia bark and essential oil of Citrus clementina clementine peel.

Rosa Canina Rosehip CO ₂ Extract (INCI: Rosa Canina (Rosehip) Fruit Extract) is a commercially available product obtained by supercritical fluid extraction with carbon dioxide.

Dry extract of Magnolia Officinalis Magnolia Bark (INCI: Magnolia Officinalis Bark Extract) is a commercially available product obtained, for example, by freeze-drying a purified hydroalcoholic extract.

According to the invention, the essential oil of clementine peel can be used. Citrus clementina is a commercially available product (for example, "Clementine Oil 5 fold MD" from VENTOS URL: https://www.ventos.com). Alternatively, an essential oil containing from about 40 to about 99 wt% limonene, from about 1 to about 10 wt% myrcene, from about 0.1 to about 1 wt% alpha pipene, from about 0 , 1 to about 1 wt.% Linalool. Preferably, the limonene content is from about 50 to about 95 wt%, preferably from about 50 to about 90 wt%, preferably from about 50 to about 85 wt%, preferably from about 50 to about 80 wt%, preferably from about 50 to about 75 wt%, preferably about 50 to about 70 wt%, preferably about 50 to about 65 wt%, preferably about 50 to about 60 wt%, preferably about 50 to about 55 wt% .%, preferably from about 40 to about 50 wt.% Preferably, the refractive index nD20 of such an essential oil is from about 1.4680 to about 1.4800.

The complex of active ingredients according to the invention contains or consists of: CO ₂ -extract of rosehip flowers Rosa Canina, dry extract of the bark of Magnolia Officinalis and essential oil of the peel Citrus clementina exhibits a synergistic effect, with a ratio of (0.05-0.5) :( 0.001 -0.1) :( 0.005-0.3) by mass. Preferably, the ratio is (0.05-0.4) :( 0.005-0.1) :( 0.01-0.2) by mass, preferably the ratio is (0.1-0.3) :( 0, 01-0.1) :( 0.02-0.1) by mass, preferably the ratio is (0.1-0.2) :( 0.01-0.05) :( 0.02-0.05 ) by weight.

An aspect of the claimed invention is a dentifrice and / or oral care product that contains a complex of active ingredients according to the invention. The complex of active ingredients according to the invention contains or consists of: CO ₂ -extract of rose hips Rosa Canina, dry extract of the bark of Magnolia Officinalis and essential oil of the peel of Clementine Citrus clementina, and exhibits a synergistic effect when contained in a product for the care of teeth and / or oral cavity in an amount of from about 0.1 to about 0.3 wt.%, preferably from about 0.15 to about 0.25 wt.%, preferably about 0.2 wt.% In a preferred embodiment, when the care agent teeth and / or oral cavity is a toothpaste, the complex according to the invention is contained in an amount of from about 0.1 to about 0.3 wt.%, preferably from about 0.15 to about 0.25 wt.%, preferably from about 0 , 16 to about 0.17 wt.%, Preferably about 0.163 wt.% In a preferred embodiment of the invention, when the dentifrice and / or oral cavity is a mouthwash, the complex of the invention is present in an amount from about 0.1 to about 0.3 wt%, preferably about 0.15 to about 0.25 wt%, preferably about 0.22 to about 0.24 wt%, preferably about 0.23 wt. %.

The dentifrice and / or oral care product of the invention may further comprise an enzyme suitable for use in dentifrice and / or oral care products, wherein the enzyme suitable for use in dentifrice and / or oral care products, is bromelain, papain, or a mixture thereof. Such enzymes are commercially available (INCI: Bromelain, Papain) and their use in dentifrice and / or oral care compositions is known (eg, SENSITIVE ULTRA Toothpaste from SPLAT, LICVUM-GEL Toothpaste from SPLAT).

The dentifrice and / or oral care product of the invention may further comprise one or more of: grapefruit essential oil such as Citrus Paradisi, peppermint essential oil such as Mentha Piperita, anise essential oil such as Illicium Verum, basil essential oil such as Ocimum Basilicum, a clove essential oil such as Eugenia Caryophyllus. These essential oils are commercially available.

The dentifrice and / or oral care product of the invention may further comprise a bacterial lysate. Such lysates are commercially available and their use in dentifrice and / or oral care compositions is known (eg EXTRA FRESH Toothpaste from SPLAT). The bacterial lysate may be a lysate of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus bulgaricus, or a mixture thereof.

Means for caring for teeth and / or oral cavity according to the invention may additionally contain one or more of: plantain extract, clover extract, Japanese Sophora extract, or a mixture thereof; for example, plantain leaf extract, clover flower extract, Japanese Sophora bud extract, or a mixture thereof; for example, Plantago Lanceolata plantain leaf extract, Trifolium Pratense clover flower extract, Japanese Sophora Japonica bud extract, or a mixture thereof; for example, water-glycerin extract of plantain leaves Plantago Lanceolata, water-glycerin extract of clover flowers Trifolium Pratense, water-glycerin extract of buds of Japanese Sophora Japonica, or a mixture thereof. Such extracts are commercially available.

The dentifrice and / or oral care product of the invention may further comprise hibiscus extract; for example, hibiscus flower extract; for example, Hibiscus Sabdariffa hibiscus flower extract; for example, an aqueous extract of hibiscus flowers Hibiscus Sabdariffa. This extract is commercially available.

Means for caring for teeth and / or oral cavity according to the invention may additionally contain an extract of badan; for example, berry root extract; for example, bergenia crassifolia bergenia root extract; for example, dry extract of bergenia crassifolia bergenia root.

Means for caring for teeth and / or oral cavity according to the invention may additionally contain pomegranate extract; for example, pomegranate fruit extract; for example, Punica Granatum pomegranate fruit extract; for example, fermented pomegranate fruit extract Punica Granatum.

An aspect of the claimed invention is a toothpaste. Means for caring for teeth and / or oral cavity according to the invention can be a toothpaste containing, wt%:

CO ₂ -extract of rose hips Rosa Canina preferably 0.1 dry extract of magnolia bark Magnolia Officinalis preferably 0.01 essential oil of clementine Citrus clementina preferably 0.05

moreover, such a toothpaste may additionally contain:

enzyme bromelain (0.001-0.2), preferably 0.04 enzyme papain (0.003-0.03), preferably 0.005 grapefruit essential oil Citrus Paradisi (0.001-0.2), preferably 0.001 peppermint essential oil Mentha Piperita (0.001-0.6), preferably 0.01 Eugenia Caryophyllus clove essential oil (0.001-0.1), preferably 0.04 water-glycerin plantain leaf extract Plantago lanceolata (0.1-1), preferably 0.3 aqueous extract of hibiscus flowers Hibiscus Sabdariffa (0.001-0.1), preferably 0.01 mixture of lysates Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus bulgaricus (0.01-1), preferably 0.07

An aspect of the claimed invention is an oral rinse. Means for caring for teeth and / or oral cavity can be a mouthwash containing, wt%:

СО ₂ -extract of rose hips flowers Rosa Canina preferably 0.2 dry extract of magnolia bark Magnolia Officinalis preferably 0.0, essential oil of clementine Citrus clementina preferably 0.02 enzyme bromelain (0.005-0.1), preferably 0.02 essential oil of anise Illicium Verum (0.001-0.005), preferably 0.003 Basil essential oil Ocimum Basilicum (0.001-0.01), preferably 0.002 Eugenia Caryophyllus clove essential oil (0.001-0.01), preferably 0.007 water-glycerin extract of clover flowers Trifolium Pratense (0.01-1), preferably 0.2 water-glycerin extract of Japanese Sophora buds Sophora Japonica (0.01-3), preferably 0.8 aqueous extract of hibiscus flowers Hibiscus Sabdariffa (0.001-0.1), preferably 0.01 fermented pomegranate fruit extract Punica Granatum (0.01-1), preferably 0.1 a mixture of lysates of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis (0.01-1), preferably 0.05

An aspect of the claimed invention is the use of a complex of active ingredients according to the invention or a dentifrice and / or oral care agent according to the invention for the prevention of oxidative stress in the oral cavity.

An aspect of the claimed invention is a method for preventing oxidative stress in the oral cavity, comprising topically applying a complex of active ingredients according to the invention or a dentifrice and / or oral cavity according to the invention.

An aspect of the claimed invention is the use of a complex of active ingredients according to the invention or a dentifrice and / or oral care agent according to the invention for reducing periodontal inflammation in a subject having such inflammation.

An aspect of the claimed invention is the use of a complex of active ingredients according to the invention or a dentifrice and / or oral care product according to the invention to reduce bleeding of the gums in a subject subject to it.

An aspect of the claimed invention is a method for reducing periodontal inflammation in a subject having such inflammation, comprising topically applying a complex of active ingredients according to the invention or a dentifrice and / or oral cavity according to the invention.

An aspect of the claimed invention is a method for reducing gingival bleeding in a subject subject, comprising topically applying a complex of active ingredients according to the invention or a dentifrice and / or oral cavity according to the invention.

An aspect of the claimed invention is a method for the prevention of inflammatory diseases after dental prosthetics when using removable dentures, comprising the topical application of a complex of active ingredients according to the invention or a means for caring for teeth and / or oral cavity according to the invention.

An aspect of the claimed invention is a method for the prophylaxis of inflammatory diseases after dental crowns have been placed, comprising topically applying a complex of active ingredients according to the invention or a means for caring for teeth and / or oral cavity according to the invention.

It was found that biologically active substances (BAS) contained in the components of the claimed composition unexpectedly exhibit a synergistic effect with a decrease in the severity of inflammation of the gums and periodontal disease as a whole, which is confirmed by the experimental data included in the present description. Combined use in one product leads to a powerful increase in anti-inflammatory and antioxidant effects, and the antibacterial effect on pathogenic microflora allows for homeostasis of the immune system in the oral cavity. A distinctive feature is that the components act in inflammation both above and below the edge of the gums, therefore, the effectiveness of treatment of periodontal diseases increases.

First, biologically active substances must penetrate into the periodontal junction and stop the growth of pathogenic microorganisms. To stop the destruction of the gums and reduce the area of inflammation, it is necessary to stop oxidative stress. Polysaccharides from rosehip flower extract quickly dissolve in saliva and penetrate into the gingival sulcus, where they are absorbed by pathogenic microorganisms. Polysaccharides inhibit the enzyme glycosyltransferase of bacteria P. gingivalis, P. intermedia, S. oralis, A. actinomycetemcomitans, interrupting the synthesis of biofilms from polysaccharide dextran around the area of gingival inflammation. The activated immune defense enzyme lysozyme destroys bacterial colonies, providing access for the following components. Lignans magnolol and honokiol from magnolia bark extract have an antibacterial effect against pathogenic bacteria P. gingivalis, F. nucleaticum, A. actinomycetemcomitans, P. intermedia, C. gingivalis, blocking the glycosyltraneferase enzyme. Prolonged release, limonene, linalool and citral from the essential oil of the clementine peel blocks the activity of elastase and collagenase, which destroy type I collagen fibers - the main protein of the extracellular matrix.

и гидроксидного радикала

. Уменьшение количества АФК приводит к прекращению перекисного окисления липидов мембран клеток, активации клеток иммунитета и разрушения сосудов местного кровотока, снижая кровоточивость, отечность десен и поступление цитокинов и провоспалительных факторов в общий кровоток. Также дельфинидин активирует дифференцировку регуляторных Т-лимфоцитов посредством выделения фактора TGF-β, который прекращает хемотаксис макрофагов и Т-лимфоцитов в зону повреждения десны. Лигнаны магнолол и хонокиол подавляют синтез провоспалительных цитокинов (TNFα, IL-1β) и метаболитов арахидоновой кислоты (простагландинов, лейкотриенов). Активные вещества эфирного масла гвоздики повышают экспрессию защитных ферментов от окислительного стресса: глутатионпероксидазы, каталазы, супероксиддисмутазы, глутатион S-трансферазы, снижая число оставшихся АФК в клетках. Совместно действуют длительно действующие флавоноиды из экстракта шиповника (гесперидин, нарингин, рутин, кверцетин) посредством торможения гиалуронидазы, поэтому повышается концентрация гиалуроновой кислоты во внеклеточном матриксе. Молекулы гиалуроновой кислоты снижают динамическую нагрузку на волокна коллагена и эластина, поэтому они быстрее восстанавливаются, увеличивая эластичность капилляров сосудов и снижая их проницаемость. Для снижения времени восстановления гидролизуемые танины (ругозин D и теллимаграндин II) из экстракта шиповника образуют плотную пленку из белков слюны, секреторного IgA и раневого экссудата за счет своего дубящего действия на воспаленных слизистых оболочках выше и ниже края десны. Образовавшаяся белковая пленка защищает клетки слизистой оболочки и нервные окончания от действия раздражающих веществ, поэтому снижается болевой порог, повышенная чувствительность зубов, отек и гиперемия слизистой. Метиловый эфир салициловой кислоты из эфирного масла гвоздики и α-бисоболол из экстракта цветков шиповника адсорбируются на поверхности белковой пленки и оказывают пролонгированное антибактериальное действие, предотвращая чрезмерное развитие патогенных микроорганизмов и повторное развитие воспаления десен.Additionally, organic acids (citric, malic, tartaric) from hibiscus extract bind free calcium ions in the gingival fluid, preventing the formation of subgingival tartar - the main factor in the development of periodontal disease. Anthocyanins (delphinidin, 3-O-sambulozide delphinidin, delphinidin-3-glucoside, etc.) from hibiscus extract and flavonoids from rosehip flower extract quickly dissolve in saliva and penetrate deeply into the inflamed areas of the gums, providing an antioxidant effect by stabilizing ROS: superoxide radical anion

and hydroxide radical

... A decrease in the amount of ROS leads to the cessation of lipid peroxidation of cell membranes, activation of immune cells and destruction of local blood vessels, reducing bleeding, gingival swelling and the flow of cytokines and pro-inflammatory factors into the general bloodstream. Delphinidin also activates the differentiation of regulatory T-lymphocytes by releasing the TGF-β factor, which stops the chemotaxis of macrophages and T-lymphocytes in the area of gingival injury. Lignans magnolol and honokiol inhibit the synthesis of pro-inflammatory cytokines (TNFα, IL-1β) and arachidonic acid metabolites (prostaglandins, leukotrienes). The active substances of clove essential oil increase the expression of protective enzymes against oxidative stress: glutathione peroxidase, catalase, superoxide dismutase, glutathione S-transferase, reducing the number of remaining ROS in cells. Long-acting flavonoids from rosehip extract (hesperidin, naringin, rutin, quercetin) work together by inhibiting hyaluronidase, therefore, the concentration of hyaluronic acid in the extracellular matrix increases. Hyaluronic acid molecules reduce the dynamic load on collagen and elastin fibers, so they recover faster, increasing the elasticity of the vascular capillaries and reducing their permeability. To reduce the recovery time, hydrolyzed tannins (rugozin D and tellimagrandin II) from rosehip extract form a dense film of saliva proteins, secretory IgA and wound exudate due to their tanning effect on the inflamed mucous membranes above and below the gum edge. The formed protein film protects the cells of the mucous membrane and nerve endings from the action of irritating substances, therefore, the pain threshold, increased sensitivity of the teeth, edema and hyperemia of the mucosa are reduced. Salicylic acid methyl ester from clove essential oil and α-bisobolol from rosehip flower extract are adsorbed on the surface of the protein film and have a prolonged antibacterial effect, preventing the excessive development of pathogenic microorganisms and the re-development of gum inflammation.

Thus, the complex of plant biologically active substances suppresses oxidative stress and inflammation, prevents the destruction of extracellular matrix proteins (collagen and elastin), reduces the number of cells of the immune system and the release of pro-inflammatory factors from them. In the course of research, it was found and confirmed that the components have antioxidant and antibacterial effects, enhancing each other's action to reduce periodontal inflammation.

Experimental data

The examples included in the present description are not limiting of the claimed invention and are provided only for the purpose of illustration and confirmation of the achievement of the expected technical results. These examples are one of many experimental data obtained by the inventors, which confirm the effectiveness of tools within the scope of the invention.

Preclinical studies have been conducted to evaluate the effectiveness of the compositions of the invention.

Example 1

To determine the antioxidant activity (AOA) of CO ₂ -extract of rose hips Rosa Canina flowers, dry extract of the bark of Magnolia Officinalis and essential oil of the peel of Clementine Citrus clementina, the potentiometric method was used due to the ease of implementation in the laboratory, rapidity and informational content for research purposes. Plant extracts are represented by a wide range of different biologically active compounds, including polyphenolic ones, therefore, one of these properties can be the integral indicator of AOA, with the help of which it is possible to evaluate the effect of using the combination in oxidative stress. Substances of a polyphenolic nature are represented by simple phenolic compounds, hydroxybenzoic acids (gallic, protocatechuic, salicylic acids), hydroxycinnamic acids (hydroxycinnamic, caffeic, ferulic acids and its metabolite curcumin), coumarins, stilbenes (resveratrol), flavonoids, an , flavones, flavonols, chalcones).

The oxidant / antioxidant balance of the body determines the redox reactions, therefore, the general principles for determining the integral antioxidant activity are based on redox reactions and can be implemented in different ways: chemical (redox titration) and physicochemical (electrochemical methods of determination). The most convenient method to use is potentiometry, since it has high sensitivity, good reproducibility of results, ease of implementation (compared to other methods), takes into account the antioxidant activity of a large spectrum of biologically active substances and the result is not recorded by the color change in the system, which makes it possible to use it to determine antioxidant activity of turbid solutions. Potentiometry is characterized by high sensitivity and rapidity, the detection limit for polyphenols and flavonoids is at the level of 10 ^-9 -10 ^-12 g, which is an advantage when determining small amounts of biologically active substances in plant extracts. The potentiometric method with a mediator system, which is adapted to the analysis of a wide range of objects, is quite informative for the assessment of antioxidant activity. The mediator is the K ₃ [Fe (CN) ₆ ] / K ₄ [Fe (CN) ₆ ] system.

Determination of antioxidant activity was carried out using a potentiometric method using potassium o-hexacyanoferrate (III) as a model of an oxidizing agent. The use of this complex compound is justified both from a thermodynamic point of view and from the point of view of obtaining an optimal analytical signal. The potential was measured after the passage of a chemical reaction between the antioxidants of the test sample and the oxidizing agent used, and the subsequent addition of the oxidizing agent. The potential change in this case occurs as a result of a chemical reaction in the solution: nFe (III) + AO = nFe (II) + AO _Ox , where AO is an antioxidant; AO _Ox - antioxidant oxidation product; n is the coefficient corresponding to the number of functional groups in the molecule that provide the antioxidant properties of the compound. In the case of polyphenolic compounds, these functional groups are hydroxyl groups - OH, conjugated with an aromatic ring.

Due to the excess of the oxidizing agent in the system, after the chemical reaction with antioxidants, the equilibrium between the oxidized form of the metal in the complex compound and the formed reduced form of the metal complex is established quickly. After introducing an extract containing antioxidants into the sample solution, the potential of the system can be expressed by the equation:

where E ₁ is the potential measured after the introduction of the first additive of the test sample, Volt; C _Ox is the concentration of the complex compound containing the oxidized form of the metal, g / l; X is the concentration of the extract in the test sample, g / l.

After the second addition of an oxidizing agent and the passage of a chemical reaction between the oxidized form of the metal in the complex and antioxidants, with the addition of a reagent, the potential can be expressed by the equation:

where E ₂ is the potential measured after the addition of the reagent additive, Volt; С ' _Ox is the concentration of the complex compound containing the oxidized form of the metal in the second additive, g / l.

Antioxidant activity is calculated using the following formula:

Since the antioxidant molecule can include several functional groups with antioxidant properties, AOA is understood as the effective equivalent concentration of antioxidants that have reacted with K ₃ [Fe (CN) ₆ ].

Toothpaste and rinse bases containing CO ₂ -extract of rose hips Rosa Canina, dry extract of magnolia bark Magnolia Officinalis and essential oil of clementine Citrus clementina were chosen as the test samples.

The weight of the sample of the base of the paste with the active ingredient was 2 g. This choice of the weight of the sample was due to the fact that 2 g is the average amount of paste that is applied by a person to a toothbrush and is used for cleaning teeth in one application.

The volume of the analyzed base of the rinse aid with the active ingredient was 10 ml. This choice of volume was due to the fact that 10 ml is the average amount of rinse a person uses for rinsing in one application.

As a reference sample, ascorbic acid was chosen, which is known for its antioxidant effects and is widely used in various products.

Titration was carried out in an electrochemical cell consisting of a platinum indicator electrode and a silver chloride reference electrode. The completion of the titration reaction was accompanied by a sharp change in the value of the potential of the electrochemical cell.

The analysis was carried out in a solution of phosphate buffer with a total volume of 100 ml at pH = 7.4 and with constant stirring. In the buffer solution containing the oxidizing agent K ₃ [Fe (CN) ₆ ] with a concentration of 5.0 g / L, an exact weighed portion of the analyzed sample was introduced, after the establishment of equilibrium in the system, the subsequent addition of the oxidizing agent was introduced. The potential was recorded with each addition of the titrant K ₃ [Fe (CN) ₆ ].

The results of determining the AOA of the studied samples of the base of the toothpaste, obtained by potentiometric methods using K ₃ [Fe (CN) ₆ ] as a model of the oxidant are presented in Table 1.

According to Table 1, it can be concluded that the antioxidant activity of the amount of biologically active substances in the base is higher than the theoretically calculated total antioxidant activity of individual active substances. The antioxidant activity of each of the components exceeds the activity of ascorbic acid by several orders of magnitude, taking into account the difference in the concentrations of substances in the solution of the electrochemical cell.

The results of determining the AOA of the studied samples of the base of the rinse, obtained by potentiometric methods using K ₃ [Fe (CN) ₆ ] as a model of the oxidant are presented in Table 2.

According to Table 2, it can be concluded that the antioxidant activity of the amount of biologically active substances in the base is higher than the theoretically calculated total antioxidant activity of individual active substances. The antioxidant activity of each of the components exceeds the activity of ascorbic acid by several orders of magnitude, taking into account the difference in the concentrations of substances in the solution of the electrochemical cell.

Consequently, to determine AOA as an integral indicator for objects with a complex matrix, one can use the potentiometric method with the oxidized form of the metal.

Example 2

In the experimental groups, patients used the claimed complex of plant extracts and essential oils in the composition of toothpaste and rinse aid, and the effectiveness of the complex was assessed using various indices. The common indicator is the oral hygiene index, since improper brushing of the teeth contributes to caries and severe inflammation of the gums.

Toothpaste with the composition shown in Table 3 was used as a base # 1 for the inclusion of components, and a mouthwash with the composition shown in Table # 4 as base # 2.

To determine the cleansing effect of a toothpaste containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 37 subjects. Patients at the 1st visit cleaned their teeth for 2 minutes, before and after cleaning, the hygiene index was measured. Then the patients applied the composition in the form of a toothpaste 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an intermediate visit to the dentist on the 14th day of the study.

To determine the cleansing effect of a mouthwash containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 40 subjects. To evaluate the effect of the rinse aid, a group of patients used the rinse aid. During the 1st visit, the patients used the mouthwash for 30 seconds, before and after the hygiene index was measured. Then the patients applied the product 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an interim visit to the dentist on the 14th day of the study.

The parameter for the overall assessment of the effectiveness of plaque removal by the composition was the hygiene index (Green-Vermillion index), determined by the dentist during the visit of the patients. A general positive trend is the gentle cleaning of teeth from dental plaque.

For all quantitative data, the group arithmetic mean (M), median (Me), standard deviation (SD) and standard error of the mean (SEM) were calculated. The results were processed using the IBM SPSS Statistics 22.0 software (StatSoft, Russia). The likelihood of differences in the mean values at different points in time was determined using the ANOVA test for dependent samples. Differences were considered significant at a significance level of p <0.05.

Results.

4 weeks after the start of the study, pronounced changes in the estimated indicator were observed. After brushing the teeth with toothpaste during 1 visit, the value of the oral hygiene index was 0.73 ± 0.36 and improved by 27.96% compared to the initial value of 1.02 ± 0.49. At the end of the study, the index value was 0.28 ± 0.12. Thus, after 4 weeks of using the toothpaste, oral hygiene improved by 85% in 50% of patients, and by 60% in the remaining 50% of patients, which indicates the pronounced cleansing properties of the test toothpaste. Considering the fact that the patients at the entrance showed a good level of oral hygiene at the entrance, the observed effect with a greater severity of gingivitis will be more noticeable, and the oral hygiene index is significantly lower.

In patients in the case of using the mouthwash on visit 2, the value of the oral hygiene index was 0.72 ± 0.32 and improved by 73.62% compared to the initial indicator of 2.71 ± 0.47. At the end of the study, the index value was 0.41 ± 0.53. Thus, after 4 weeks of use, oral hygiene improved by 85.06%, which indicates the pronounced cleansing properties of the studied mouthwash. Thus, after 4 weeks of using the rinse in 80% of patients, oral hygiene improved by 97%, and in the remaining 20% of patients - by 80%, which indicates the pronounced cleansing properties of the rinse (Table 5).

The use of paste and rinse with the claimed complex led to effective cleaning of tooth enamel, which indicates the removal of most of the plaque, which is a causal factor in the development of inflammation of the gums and the oral cavity in general.

Since the compositions contained rosehip flower extract containing water-soluble polysaccharides and magnolia bark extract containing lignans magnolol and honokiol, with a synergistic antibacterial effect due to blocking the glycosyltransferase enzyme, long-term use decreased the number of pathogenic microorganisms in the oral cavity. The base of the composition cleared soft plaque and increased the access of biologically active substances to hard-to-reach periodontal joints, thereby increasing the effectiveness of biologically active substances against pathogenic bacteria P. gingivalis, F. nucleaticum, A. actinomycetemcomitans, P. intermedia, C. gingivalis. Inhibition of the glycosyltransferase enzyme reduced the amount of plaque formed by bacteria, thereby increasing the cleansing effect of the composition and influencing the anti-inflammatory effect with prolonged use.

Example 3

In the experimental groups, patients used the claimed complex of plant extracts and essential oils in the composition of toothpaste and rinse aid, and the effectiveness of the compositions was evaluated using the PMA index. Since gingivitis and periodontitis are inflammatory diseases of the gums and gingival junction, reducing inflammation will help reduce the severity of pain and improve the patient's quality of life as a whole.

Toothpaste with the composition shown in Table 3 was used as a base No. 1 for the inclusion of components, and a mouthwash with the composition shown in Table No. 4 as base No. 2.

To determine the anti-inflammatory effect of a toothpaste containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 37 subjects. The PMA index was measured in patients during 1 visit. Then the patients applied the composition in the form of a toothpaste 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an intermediate visit to the dentist on the 14th day of the study.

To determine the anti-inflammatory effect of a mouthwash containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 40 subjects. The patients used a mouthwash. The PMA index was measured in patients during 1 visit. Then the patients applied the product 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an interim visit to the dentist on the 14th day of the study.

The parameter for assessing the anti-inflammatory effect of the composition was the PMA index, determined by the dentist during the visit of the patients. The overall positive trend is a decrease in gum inflammation.

For all quantitative data, the group arithmetic mean (M), median (Me), standard deviation (SD) and standard error of the mean (SEM) were calculated. The results were processed using the IBM SPSS Statistics 22.0 software (StatSoft, Russia). The likelihood of differences in the mean values at different points in time was determined using the ANOVA test for dependent samples. Differences were considered significant at a significance level of p <0.05.

Results.

4 weeks after the start of the study, pronounced changes in the estimated indicator were observed. After brushing the teeth with paste during the first 14 days, the PMA index was 22.26 ± 9.57, which is 32.27% less than the initial value of 33.46 ± 8.91. At the end of the study, the index value was 2.28 ± 4.52. Thus, after 4 weeks of use, gum inflammation decreased by 94.73%, which indicates a pronounced anti-inflammatory effect of the composition.

In the case of using the rinse aid during the 2nd visit, the PMA index value was 25.37 ± 6.36 and improved by 41.32% compared to the initial value of 43.23 ± 6.25. At the end of the study, the index value was 13.05 ± 8.20. Thus, after 4 weeks of use, gum inflammation decreased by 69.81%, which indicates a moderate anti-inflammatory effect of the studied mouthwash (Table 6).

The use of a paste and a rinse aid with the claimed complex led to an effective reduction in the severity of inflammation, which indicates an anti-inflammatory effect in such diseases of the oral cavity as gingivitis and periodontitis.

The base of the compositions cleared soft plaque and increased the access of biologically active substances to hard-to-reach periodontal joints, thereby increasing the effectiveness of biologically active substances. Since the compositions contained a whole complex of biologically active substances: rosehip flower extract containing polyphenolic compounds; magnolia bark extract containing the lignans magnolol and honokiol; Clementine essential oil, containing terpenoids, flavonoids, increases the expression of protective enzymes glutathione peroxidase, catalase, superoxide dismutase, glutathione S-transferase during oxidative stress, contributing to the prolonged restoration of the redox balance in the gum tissues. Reducing the severity of inflammation helps to reduce tooth sensitivity, bleeding and improve the quality of life of patients in general.

Example 4

In experimental groups, patients used the claimed complex of plant extracts and essential oils in the composition of toothpaste and rinse, and the effectiveness of the compositions was evaluated using the SBI bleeding index according to Muhlemann and Son. Since gingivitis and periodontitis are inflammatory diseases of the gums with visible bleeding, reducing the bleeding of the gums will help stop the inflammation and improve the patient's overall quality of life.

Toothpaste with the composition shown in Table 3 was used as a base # 1 for the inclusion of components, and a mouthwash with the composition shown in Table # 4 as base # 2.

To determine the effect of a toothpaste containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 37 subjects. In patients at visit 1, the SBI bleeding index was measured according to Muhlemann and Son. Then the patients applied the composition in the form of a toothpaste 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an intermediate visit to the dentist on the 14th day of the study.

To determine the effect of a mouthwash containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 40 subjects. In patients during 1 visit, the SBI bleeding index was measured according to Muhlemann and Son. Then the patients applied the product 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an interim visit to the dentist on the 14th day of the study.

The parameter for assessing the hemostatic and, in general, anti-inflammatory effect of the composition was the gingival bleeding index SBI according to Muhlemann and Son, determined by the dentist during the visit of the patients. A general positive trend is a decrease in gum bleeding.

For all quantitative data, the group arithmetic mean (M), median (Me), standard deviation (SD) and standard error of the mean (SEM) were calculated. The results were processed using the IBM SPSS Statistics 22.0 software (StatSoft, Russia). The likelihood of differences in the mean values at different points in time was determined using the ANOVA test for dependent samples. Differences were considered significant at a significance level of p <0.05.

results

4 weeks after the start of the study, pronounced changes in the estimated indicator were observed. After brushing the teeth with paste during the first 14 days, the SBI index was 0.38 ± 0.49, which is 36.36% less than the initial value of 33.46 ± 8.91. At the end of the study, the index value was 0.00 ± 0.00. Thus, after 4 weeks of use, the bleeding of the gums completely disappeared, that is, it decreased by 100.00%, which indicates a significant hemostatic effect of the studied toothpaste.

When the mouthwash was used on visit 2, the SBI value was 1.35 ± 0.49 and improved by 56.45% compared to the initial value of 3.10 ± 0.85. At the end of the study, the index value was 0.24 ± 0.43. Thus, after 4 weeks of use, the bleeding of the gums decreased by 92.26%, which indicates a significant hemostatic effect of the investigated rinse (Table 7).

The use of a paste and rinse with the claimed complex led to an effective reduction in bleeding, which indicates a hemostatic effect in inflammatory diseases of the oral cavity, for example, gingivitis and periodontitis.

и гидроксидного радикала

) при оксидативном стрессе, то при длительном применении снижалась степень воспаления полости рта.The base of the compositions cleared soft plaque and increased the access of biologically active substances to hard-to-reach periodontal joints, thereby increasing the effectiveness of biologically active substances. Since the compositions contain a whole complex of biologically active substances with a synergistic anti-inflammatory effect due to the stabilization and neutralization of ROS (superoxide anion radical

and hydroxide radical

) with oxidative stress, then with prolonged use, the degree of inflammation of the oral cavity decreased.

One of the manifestations of gum inflammation is their bleeding, therefore, to reduce the severity of inflammation, it is necessary to use compositions with a hemostatic effect. Long-acting and highly active flavonoids of rosehip extract (hesperidin, naringin, rutin, quercetin) reduce the activity of hyaluronidase at the site of inflammation, helping to restore hyaluronic acid molecules in the gum tissue. Limonene from the essential oil of clementine peel blocks the activity of collagenase and elastase, which break down the fibers of type I collagen and elastin, which binds collagen into the matrix. Hyaluronic acid molecules reduce the dynamic load on the fibers of the extracellular matrix, contributing to the restoration of collagen in the media of blood vessels and the elasticity of vascular capillaries in the gums. Hydrolyzed tannins from rosehip extract provide a prolonged hemostatic and generally anti-inflammatory effect through the formation of a dense protective film of saliva proteins, secretory IgA and wound exudate above and below the gingival margin. The formed film prevents the action of pathogenic microorganisms, creates conditions for the restoration of blood vessels and inflamed tissue, reduces edema and erosion of the gum epithelium.

Example 5

In the experimental groups, patients used the claimed complex of plant extracts and essential oils in the composition of toothpaste and rinse aid, and the effectiveness of the composition was assessed using the index of intensity of tooth hyperesthesia (according to Shtorina). Since gingivitis and periodontitis are inflammatory diseases of the gums with increased pain sensitivity to various stimuli (temperature, chemical, tactile), a decrease in gum sensitivity will improve the course of inflammatory diseases of the oral cavity and the patient's quality of life as a whole. A general positive trend is a decrease in the sensitivity of sore gums and teeth.

Toothpaste with the composition shown in Table 3 was used as a base No. 1 for the inclusion of components, and a mouthwash with the composition shown in Table No. 4 as base No. 2.

To determine the effect of a toothpaste containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 37 subjects. At the 1st visit, patients cleaned their teeth for 2 minutes, before and after cleaning, the index of the intensity of tooth hyperesthesia was measured. Then the patients applied the composition in the form of a toothpaste 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an intermediate visit to the dentist on the 14th day of the study.

To determine the effect of a mouthwash containing a complex of extracts and essential oils, a single-center, prospective, non-randomized study was conducted in 40 subjects. During the 1st visit, the patients used the mouthwash for 30 seconds, before and after the index of the intensity of tooth hyperesthesia was measured. Then the patients applied the product / products 2 times a day for 2 minutes, in the morning and in the evening, for 4 weeks with an intermediate visit to the dentist on the 14th day of the study.

The parameter for the desensitive effect of the composition was the index of the intensity of dental hyperesthesia, determined by the dentist during the visit of the patients. The general positive trend is a decrease in the sensitivity of the teeth and gums.

For all quantitative data, the group arithmetic mean (M), median (Me), standard deviation (SD) and standard error of the mean (SEM) were calculated. The results were processed using the IBM SPSS Statistics 22.0 software (StatSoft, Russia). The likelihood of differences in the mean values at different points in time was determined using the ANOVA test for dependent samples. Differences were considered significant at a significance level of p <0.05.

Results.

The assessment of the prevalence of tooth hypersensitivity by the Shtorina index (IHRI) revealed that the study product reduces tooth sensitivity after the first use. After brushing the teeth with the composition at 1 visit, the value of the oral hygiene index was 2.22 ± 1.75 and improved compared to the initial indicator of 2.53 ± 1.66. At the 2nd visit to the dentist, it was found that the overall sensitivity decreased by 23.55%. At the end of the study, the index value was 0.45 ± 0.23. Thus, after 4 weeks of using the toothpaste in 70% of patients, the sensitivity of the teeth decreased by 92%, and in the remaining 30% of patients - by 60%, which indicates a pronounced desensitive effect of the test product. Desensitization of teeth is a long-term process due to prolonged remineralization of enamel and prolonged penetration of active substances into the dentinal tubules of the tooth.

In the case of using the rinse aid on visit 2, the value of the Shtorina index was 0.05 ± 0.22 and improved by 92.31% compared to the initial indicator of 0.65 ± 0.14. Further, in the course of the clinical study, the index of the intensity of dental hyperesthesia remained constant, i.e. the sensitivity of the teeth has decreased to normal levels. At the end of the study, the index value was 0.05 ± 0.22. Thus, after 4 weeks of using the rinse in 95% of patients, tooth sensitivity decreased by 100%, and in the remaining 5% of patients - by 80%, which indicates a pronounced desensitive effect of the studied rinse. The high value of the indicator for assessing the desensitive effect can be explained by the greater penetration of the rinse into hard-to-reach, inflamed places (Table 8).

The use of a paste and rinse with the claimed complex led to an effective reduction in tooth sensitivity, which indicates an anti-inflammatory effect in oral diseases such as gingivitis and periodontitis. The products reduced the sensitivity of the teeth to normal.

и гидроксидного радикала

) при оксидативном стрессе, то при длительном применении снижалась степень воспаления полости рта.The base of the compositions cleared soft plaque and increased the access of biologically active substances to hard-to-reach periodontal joints, thereby increasing the effectiveness of biologically active substances. Since the compositions contain a whole complex of biologically active substances with a synergistic anti-inflammatory effect due to the stabilization and neutralization of ROS (superoxide anion radical

and hydroxide radical

) with oxidative stress, then with prolonged use, the degree of inflammation of the oral cavity decreased.

One of the manifestations of gum inflammation is their increased sensitivity to various stimuli, therefore, to reduce the severity of inflammation, it is necessary to use compositions with a desensitive effect. Long-acting and highly active flavonoids and polysaccharides of rosehip extract (hesperidin, naringin, rutin, quercetin) have antioxidant and anti-inflammatory effects in the gum tissue. Lignans magnolol and honokiol inhibit the synthesis of anti-inflammatory cytokines (TNFα, IL-1β) and eicosanoids (prostaglandins PgE ₂ , PgF ₂ , leukotrienes LTC ₄ , LTB ₄ ), which have the ability to increase the activity of painful C-fibers and reduce their activation threshold to stimuli. Hydrolyzed tannins from rosehip extract provide a prolonged anti-inflammatory effect by forming a dense protective film of salivary proteins, secretory IgA and wound exudate above and below the gingival margin. The formed film prevents the action of pathogenic microorganisms, creates conditions for the restoration of blood vessels and inflamed tissue, reduces edema and erosion of the gum epithelium. This protein film also protects the cells of the mucous membrane and nerve endings from irritants, therefore, the pain threshold and increased sensitivity of the teeth are reduced.

Claims (15)

1. A complex of active ingredients for use in dental and / or oral care products, consisting of: CO ₂ -extract of rosehip flowers Rosa Canina , dry extract of Magnolia Officinalis magnolia bark and essential oil of Citrus clementina peel, taken in the ratio (0 , 05-0.5) :( 0.001-0.1) :( 0.005-0.3) by mass. 2. Means for caring for teeth and / or oral cavity containing a complex of active ingredients according to claim 1 in an amount of 0.1 to 0.3 wt.%. 3. An agent according to claim 2, further comprising an enzyme suitable for use in a dentifrice and / or oral cavity. 4. An agent according to claim 3, wherein the enzyme suitable for use in the dentifrice and / or oral cavity is bromelain, papain, or a mixture thereof. 5. The agent according to claim 2, further comprising one or more of: Citrus Paradisi grapefruit essential oil, Mentha Piperita peppermint essential oil, Illicium Verum anise essential oil, Ocimum Basilicum essential oil, Eugenia Caryophyllus clove essential oil. 6. The agent according to claim 2, further comprising a bacterial lysate. 7. The agent according to claim 6, wherein the bacterial lysate is a lysate of Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus bulgaricus, or a mixture thereof. 8. The agent according to claim 2, additionally containing one or more of: water-glycerin extract of leaves of plantain Plantago Lanceolata , water-glycerin extract of clover flowers Trifolium Pratense , water-glycerin extract of buds of Japanese Sophora Sophora Japonica, or a mixture thereof. 9. The agent according to claim 2, further comprising an aqueous extract of hibiscus flowers Hibiscus Sabdariffa . 10. The agent according to claim 2, further comprising a dry extract of Bergenia Crassifolia bergenia root. 11. The agent according to claim 2, further comprising a fermented extract of the fruit of the pomegranate Punica Granatum . 12. The agent according to claim 2, which is a toothpaste or mouthwash. 13. The use of a complex of active ingredients according to claim 1 or a means for caring for teeth and / or oral cavity according to any one of claims. 2-12 for the prevention of oxidative stress in the oral cavity. 14. The use of a complex of active ingredients according to claim 1 or a means for caring for teeth and / or oral cavity according to any one of claims. 2-12 to reduce periodontal inflammation in a subject having such inflammation. 15. The use of a complex of active ingredients according to claim 1 or a means for caring for teeth and / or oral cavity according to any one of claims. 2-12 to reduce bleeding gums in a subject subject.