RU2619338C1 - Method for preparation of composition for periodontal disease treatment - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: plant raw materials are crushed, sequential extraction from raw materials with 70% and 40% ethyl alcohol and total extraction volume ajustment to 1 liter by 40% ethyl alcohol is performed. A mixture of juniper fruit, cherry fruit and ginger rhizomes are used as raw materials at the following ratio (parts by weight): juniper fruit 4.0; cherry fruit 3,0; ginger rhizomes 2.0.

EFFECT: invention facilitates effective treatment of inflammatory periodontal diseases.

7 dwg, 5 tbl, 5 ex

Description

The invention relates to pharmaceutical technology and dentistry, in particular to a method for producing a composition for the treatment of periodontal diseases.

Known solution for topical application "Maraslavin" [http://www.rlsnet.ru/tn_index_id_2007.htm], which includes a decoction:

wormwood of Pontic herb (Artemisiaa pontica L.), savory herb (Satureja hortensis L.), clove, buds (Syzygium aromaticum (L.) Merr.et. LMPerry), black pepper (Piper nigrum L.), ginger rhizome (Zingiber officinale Roscoe). The disadvantages of this drug is the low effectiveness of maraslavin - a course of treatment of 10 or more days, unreasonably complex and expensive composition of the starting products.

There is a remedy "Phytodent" (RU No. 2147223, publ. 10.04.2000). It consists of 22 components. The disadvantages of this drug are the complexity of its standardization, the proximity of the pharmacological action of biologically active substances of the components of the composition, irrational production technology.

The closest solution to the claimed invention is a method of obtaining funds for the treatment of periodontal disease (RU No. 2259206, publ. 08/27/2005 ), including grinding plant materials, sequential extraction of raw materials with 70%, then 40% ethanol, followed by addition essential oils to combined extracts. The plant material is crushed to a particle size passing through a sieve with a diameter of 2 mm, extraction is carried out by percolation at room temperature with a ratio of raw materials: extractant 1: 6.7-10, before the process of percolation, the raw materials are insisted with 70% ethanol, use as raw materials (parts by weight): bird cherry fruits 2.0-3.0, horseradish roots 4.0-5.5, clove buds 2.0-2.5, ginger rhizome 1.5-2.5, to the combined 5-15 ml of tea tree essential oil is added to the extract and the volume is adjusted to 1 liter with 40% ethanol.

The disadvantage of this solution is the use of plant materials with intersecting pharmacological properties, as well as the addition to the technology of the stage of introducing an additional amount of separately obtained or purchased essential oil to enhance the antibacterial properties of the composition.

The objective of the invention is to develop a simpler and more effective composition for the treatment of periodontal diseases, expanding the range of existing targeted funds.

The problem is solved by the composition for the treatment of periodontal disease by sequential percolation of 70%, and then 40% ethanol of the fruits of juniper, bird cherry, ginger rhizomes in the following ratio of the weight parts of raw materials of medicinal and aromatic plants:

Juniper fruit 4.0

Cherry fruits 3.0

Ginger Rhizomes 2.0

The technical result obtained by carrying out the invention is expressed in simplifying the composition with preserving the therapeutic effect by introducing into the composition of the raw cones of juniper ordinary, containing a significant amount of biologically active substances (essential oils, flavonoids), with proven antiseptic and anti-inflammatory, as well as antioxidant effects. At the same time, the effect of tannins of bird cherry ordinary and immunostimulating, as well as the anti-inflammatory effect of biologically active substances of ginger, is preserved.

Juniper fruits are known to contain a large amount of essential oil of the composition: α-pinene (33.0 - 35.0%), β-pinene (13.0%), β-myrcene (28%), limonene (4.0- 10.0%), sabinen (1.0%), γ-muurolen (4.0%), camphene (3.6%), terpineol, α-terpineol, α-terpinene, γ-terpinene, terpinolene (0, 5 - 0.8%), α-thujen, Δ3-karen, α-terpinolene, etc. In addition, they contain a large number of polyphenolic substances (flavonoids) (Novikov, OO Juniper: phytochemistry and pharmacology of the genus Juniperus L / O.O. Novikov, D.I. Pisarev, E.T. Zhilyakova [et al.] // M.: Publishing House of RAMS, 2014 .- 178 p.).

The fruits of bird cherry contain a complex of biologically active substances of polyphenolic nature (gallic acid, hexaoxidefenic acid, tannins), which have a wide spectrum of antimicrobial and fungicidal action; flavonoid glycosides with a pronounced antioxidant, anti-inflammatory, regenerating and capillary effect, which has a beneficial effect on periodontal tissues http://www.fito.nnov.ru/special/glycozides/dube/padus_avium/).

Ginger rhizomes contain 2-3% of the essential oil, the main component of which is zingibern (up to 70%), zingiberon and others, derivatives of salicylic acid (http://flora.dobro-est.com/imbir-zingiber-opisanie-vidyi- i-vyirashhivanie-imbirya.html). They have proven antimicrobial and anti-inflammatory properties. In addition, ginger has pronounced immunomodulating properties.

On the basis of the developed composition, a water-alcohol extract is obtained by percolation method. The use of the percolation method makes it possible to more fully recover the entire complex of biologically active substances due to the higher concentration gradient that is created when the pure extractant is passed through the feed.

In the first stage, 70% ethanol is used to more fully extract substances of a terpenoid nature (essential oils) when infused, at room temperature for 24 hours.

Subsequent percolation is carried out sequentially with 70% and 40% ethanol. 40% ethanol allows exhaustively extracting biologically active substances of a polyphenolic nature (flavonoids) and their glycosylated derivatives.

The invention is illustrated by the following examples.

Example 1. To obtain the claimed funds use the fruits (cones) juniper, cherry fruits, ginger rhizomes in the following ratio of components (parts by weight):

Juniper fruit 4.0 Cherry fruits 3.0 Ginger Rhizomes 2.0

The composition is prepared as follows. 40 g of juniper fruit, 30 g of bird cherry fruit and 20 g of ginger rhizomes are crushed to a particle size of 2.0 mm in diameter and filled with 100 ml of 70% ethyl alcohol and infused for 4 hours, then the mixture is placed in a percolator, compacted, filled with 70% ethyl alcohol to the "mirror" and incubated at room temperature for a day. Then percolation is carried out: a continuous flow of extractant is passed through the percolator, and percolation is carried out sequentially first with 70% (400 ml) and then with 40% ethanol (400 ml). The resulting extracts are combined. The volume of the product obtained is adjusted to 1 liter with 40% ethyl alcohol.

The finished product is a clear brown liquid with a pleasant smell of essential oils and a bitter-spicy taste.

Example 2. The study of the antimicrobial activity of the composition.

In the study of the antibacterial action, the reference strains of microorganisms recommended by WHO to test the sensitivity to antibacterial drugs were used as test cultures:

1) Staphylococcus aureus - pcs. ATCC 25923;

2) Pseudomonas aeruginosa - pcs. ATCC 27853;

3_ E. coli - pcs. ATCC 25922;

4) Proteus vulgaris - pcs. "H" 4636.

All test cultures were grown on solid nutrient media for 24 hours at a temperature of 37 ^° C. For the study, we took only those cultures that were characterized by homogeneity of the colonies, typical growth and inherent in this type of microorganism attitude to gram stain. The following nutrient media were used for the cultivation of test microbes: the main ones used for growing clostridia, hay and Escherichia coli and Proteus, and “hungry” nutrient media for the lower layer. The study of the antibacterial activity of the developed composition (sample No. 1) was carried out in comparison with an agent for the treatment of periodontal diseases (RF patent No. 2259206) (sample No. 2) by agar diffusion. Culture of test microbes was introduced into the nutrient medium in accordance with the optical turbidity standard GISK them. Tarasevich. For each type of microorganism, a specific microbial load of microbial cells per 1 ml of medium was used: staphylococcus - 40x10 ⁶ , Pseudomonas aeruginosa - 50x10 ⁷ ; E. coli - 50x10 ⁷ ; Proteus - 50x10 ⁷ . Data on the study of antibacterial action against gram-positive and gram-negative anaerobic and aerobic microorganisms are presented in table 1 (Fig. 1).

As can be seen from the data presented, the studied samples had inhibitory properties to varying degrees with respect to both gram-positive and gram-negative microorganisms. The developed composition had a stronger inhibitory effect on growth by the microorganism than the reference drug, which practically did not affect the reproduction of Pseudomonas aeruginosa and Escherichia coli.

Example 3. The study of anti-inflammatory activity of the composition.

The anti-inflammatory activity of the composition was investigated in 40 mice. All animals were divided into 4 groups: two control and two experimental (10 animals each).

The method used in the work (Tyutyunnik V.I., Ponomareva N.G., Krivoshein Yu.S. Antimicrobial effect of essential oils isolated from plants // Cultivation and processing of essential oil crops. - Simferopol. - 1980. - C. 63-67 .), in which 0.05 ml of turpentine solution in liquid paraffin was injected intradermally to reproduce the inflammatory focus, and 0.03 ml of a 1% solution of strychnine nitrate was introduced into the necrotic lesion 24 hours after the administration of turpentine. This dose is fatal for mice, however, with this method of strychnine administration only individual animals die, since the inflammatory shaft prevents the entry of poison into the blood. In the case of the action of any anti-inflammatory drug, the barrier function is impaired and the animals die.

The animals of the first and second experimental groups were injected intramuscularly every 48 hours with 0.1 ml of an agent for the treatment of periodontal diseases (RF patent No. 2259206) (experimental group No. 1) and the developed composition (experimental group No. 2), respectively, all three injections were made, the latter - a day before the initiation of the inflammatory focus.

Instead of the developed composition, the animals of the control group No. 1 were injected with a mixture of hydrocortisone and vitamin B ₆ . Hydrocortisone and vitamin B ₆ were administered to mice twice 30 minutes before the administration of turpentine and after 6 hours, 0.025 and 0.5 μg, respectively.

Animals of the control group No. 2 were injected with 50% turpentine solution and 1% strychnine nitrate solution.

The research results are presented in table 2 (Fig. 2).

The data presented indicate that the developed composition exceeds the anti-inflammatory activity of the comparison drug and approaches the anti-inflammatory activity of hydrocortisone in combination with vitamin B ₆ .

Example 4. The study of antioxidant activity in vitro.

To study the antioxidant activity of the drugs, we used an adrenaline autooxidation system in an adrenochrome in an alkaline medium (Methods for assessing the antioxidant powers of physiologically active spoils with completely radical in vitro processes (method. Recommendations) / J.I. Gubsky]. Ministry of Health of Ukraine, 2002. - 26 p.). A solution of adrenaline was prepared in bidistilled water, only pure crystalline adrenaline was used. An exact weighed adrenaline (40.5 mg) was added to a 100 ml volumetric flask and 50 ml bidistilled water was added, after which 0.5 ml of 0.1N HCl was adjusted, the pH was adjusted to 2.0, then the volume was adjusted to bidistillate to 100 ml. The working volume was adjusted to 100 ml. 2 ml of 0.15 M carbonate buffer pH 10.2 containing 500 mg Trilon B per 1 liter of volume was added to the cuvettes of a spectrophotometer 10 mm thick. After that, the developed composition or comparison drug was introduced - an agent for treating periodontal diseases (RF patent No. 2259206) (control No. 2) with a volume of up to 0.5 ml. The reaction was started by adding 0.4 ml of adrenaline solution. The optical density was determined at a wavelength of 480 nm against a carbonate buffer. Antioxidant activity (AOA) was calculated by the degree of inhibition of autooxidation of adrenaline into adrenochrome by the formula:

AOA = (E ₀ - E _k ) / E ₀ H 100%,

where E ₀ is the optical density of the control solution, E _to is the optical density of the test solution.

Control No. 1 was a mixture of reagents without the studied compounds.

The results are presented in table 3 (Figure 3).

Thus, the highest AOA was found in the developed composition.

Example 5. The study of the effectiveness of the developed composition in dental practice.

The composition was tested on 24 patients aged 19 to 28 years with chronic catarrhal gingivitis of mild to moderate severity, who applied for help to the Interregional Center for Dental Innovations of Belgorod State University. To make a diagnosis and determine the effectiveness of the proposed composition, clinical research methods were used, such as medical history and examination, and additional ones - a simplified Green-Wermillion UIGR hygiene index, PMA gingivitis index, microcirculation assessment in periodontal tissues using laser Doppler flowmetry (LDF) on an analyzer capillary blood flow "LAK-2".

The distribution of patients by registered nosologies and by age categories is shown in table 4 (Fig. 4).

Patients initially complained of redness and swelling of the gums, itching, bad breath. The average value of the UIGR index is 4.4 ± 0.52, which indicates poor oral hygiene, the PMA index is 42 ± 0.86%.

The treatment plan included the following steps: correction of individual oral hygiene, professional brushing. Then appointed the claimed composition in the form of rinses (dilution with water 1:10). The course of treatment was 7 days. All patients received positive treatment results, starting from the third day of application of the composition. Against the background of the treatment, a trend was observed to decrease the PMA index to 23 ± 0.55%. In addition, 87% of patients noted a decrease in halitosis, lack of discomfort. An objective examination observed a decrease in hyperemia of the oral mucosa already on the 2nd day, patients noted a decrease in the intensity of bleeding gums when brushing their teeth on the 8th day.

The study of capillary blood flow in the gum tissue was carried out in the position of the patient while sitting in the dental chair after ten minutes of rest using the LACK-2 device manufactured by NPP Lazma, Moscow. The microcirculation index (PM) is one of the leading ones and represents an integral characteristic of capillary blood flow and depends on the average speed of red blood cells, the capillary hematocrit and the number of functioning capillaries.

The results of monitoring periodontal hemomicrocirculation indicators during the treatment of chronic catarrhal gingivitis are presented in table 5 (Fig. 5).

Thus, in patients with chronic generalized catarrhal gingivitis of mild severity, according to LDF, an increase in intravascular resistance, a slight decrease in vascular tone. The appearance of LDH-grams of a patient with mild chronic generalized catarrhal gingivitis is presented in (Fig. 6).

In subjects with chronic generalized catarrhal gingivitis of moderate severity, an increase in intravascular resistance, a decrease in vascular tone, an increase in the contribution of respiratory and cardiac blood flow fluctuations in the provision of the microvasculature were noted. The described processes are presented on the LDG-gram of a patient with chronic generalized catarrhal gingivitis of moderate severity (Fig. 7).

7 days after application of the composition, the microcirculation index (PM) decreased in patients of group I to 3.5 ± 0.11, which indicates the absence of signs of inflammation in the gums, and in patients of group II there was also a tendency to stop the inflammatory process.

Thus, the claimed composition for the treatment of inflammatory periodontal diseases allows to achieve the goal of the present invention, namely, a simple and effective composition for the treatment of inflammatory periodontal diseases has been developed, the range of existing target funds has been expanded.

Claims (2)

A method of obtaining a composition for the treatment of inflammatory periodontal diseases, including grinding plant materials, sequential extraction of 70% and 40% ethyl alcohol from raw materials and bringing the total volume of extraction to 1 liter with 40% ethyl alcohol, characterized in that as raw materials use a mixture of juniper fruits, cherry fruits and rhizomes of ginger in the following ratio of components, parts by weight: Juniper fruit 4.0 Cherry fruits 3.0 Ginger Rhizomes 2.0

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