RU2708260C1 - Pharmaceutical composition for preventing and treating thyroid hypothyreosis - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.

SUBSTANCE: invention refers to pharmaceutical industry, namely to a pharmaceutical composition for preventing and treating thyroid hypothyreosis. Pharmaceutical composition for preventing and treating hypothyreosis of thyroid gland in tablet form, comprising white cinquefoil and target additives, wherein the white cinquefoil is used in the form of a dry extract of a white cinquefoil, which is made by grinding 3–5-year-old cultivated roots and rhizomes with a degree of grinding of 3–5 mm, triple extraction with 40 % ethyl alcohol while heating 65–70 °C and continuous stirring, hydromodulus 1:15, concentration, extraction and drying at temperature not higher than 60 °C, and target additives are calcium stearate, croscarmellose, microcrystalline cellulose and lactose monohydrate taken in certain amounts.

EFFECT: disclosed composition is safe and effective for preventing and treating thyroid hypothyreosis.

1 cl, 7 dwg, 13 tbl

Description

The invention relates to the pharmaceutical industry, namely, to drugs for the treatment and prevention of thyroid diseases.

Currently, for the treatment of hyperthyroid and hypothyroid states, mainly synthetic thyrostatic drugs are used, which suppress or stimulate the secretory activity of the thyroid gland.

Known therapeutic and cosmetic composition for external use [RU 2314120, C1, A61K 31/198, A61K 38/24, A61K 9/20, A61P 5/14, A61P 5/06, 01/10/2008], which can be used in complex therapy and the prevention of thyroid diseases, and includes in an effective amount biologically active extracts of cinquefoil, common flax and middle stellate, taken in the ratio from (1: 1, 3: 4, 4) to (1: 1, 2: 2, 8).

The disadvantage of this technical solution is the relatively low efficiency of the application.

Also known is a biologically active food supplement for normalizing thyroid function [RU 2446815, C1, A61K 36/73, 04/10/2012], containing the roots and rhizomes of the white cinquefoil or the terrestrial part of the white cinquefoil, or their mixture, as well as echinacea purpurea and kelp in a certain ratio of components.

The disadvantage of this technical solution is also the relatively low efficiency of the application.

In addition, it is known a tool for the treatment and prevention of the thyroid gland [RU 2351354, C1, A61K 36/73, 04/10/2009], which is a collection consisting of white cinquefoil, licorice roots, grass of a series of tripartite and thallus saccharosa in the following ratio components 20: 5: 8: 7, respectively, while it contains the roots and rhizomes of the cinquefoil white or the ground part of the white cinquefoil, or a mixture thereof and can be made in the form of crushed vegetable powder, water-alcohol tinctures, liquid and dry extracts.

The tool has a wide range of therapeutic effects, it is used for the treatment and prevention of thyroid diseases, including: hypothyroidism, hyperthyroidism (Bazedov-Graves disease, nodular toxic goiter), euthyroid goiter, autoimmune thyroiditis.

The disadvantage of this technical solution is the relatively low efficiency of use for the prevention and treatment of thyroid diseases.

The closest in technical essence to the proposed is a composition for internal use in the form of tablets or capsules to normalize the function of the thyroid gland [RU 2011109721, A, A61K 36/36, 09/20/2012], including white cinquefoil and medium asterisk in the form of crushed grass, roots, rhizomes and dry extract, as well as vitamin E in the following components, wt. %:

shredded rhizomes and roots of the white cinquefoil 21-58 crushed grass of a white cinquefoil 20-64 dry cinquefoil extract 2-9 chopped medium stellate grass 4-16 vitamin e 2-3

The disadvantage of the closest technical solution is the relatively low efficiency of use and the relatively high level of undesirable side effects caused by the presence in addition to white cinquefoil other components that can cause an undesirable body reaction in individual patients.

The objective of the present invention is to develop a solid dosage form for oral administration based on a biologically active substance in order to create a pharmaceutical composition in the form of tablets for the treatment and prevention of thyroid diseases, which meets all the requirements of the Pharmacopoeia of the Russian Federation of the XIII edition and which excludes or, at least as a result, the disadvantages of the known composition are reduced on the basis of the use of the active substance of plant origin and the manufacture of its expression in the form of tablets.

The required technical result is to expand the arsenal of funds used for the prevention and treatment of thyroid diseases, increase the effectiveness of prevention and treatment and to reduce the possibility of side effects in the treatment of thyroid diseases.

The problem is solved, and the required technical result is achieved by the fact that, in the pharmaceutical composition for the prevention and treatment of thyroid diseases, made in the form of tablets, and including white cinquefoil as a therapeutically effective active substance and target additives, according to the invention, white cinquefoil as the therapeutically effective active substance is included in the form of a dry extract of white cinquefoil, made by grinding 3-5 - year-old cultivated root plants and white rhizomes of a white cinquefoil with a degree of grinding of 3-5 mm, three times extraction with 40% ethyl alcohol with heating at 65-70 ° С and constant stirring, hydromodule 1:15, extraction concentration and drying at a temperature not exceeding 60 ° С, and as target additives used calcium stearate, croscarmellose and microcrystalline cellulose and lactose monohydrate in the following components, wt. %:

cinquefoil extract 42-43 microcrystalline cellulose 26-27 lactose monohydrate 23-24 croscarmellose 5 calcium stearate 1-2

The drawing shows:

in FIG. 1, a, 1, b - thyroid gland of an intact rat (typical follicular structure of the gland, cubic thyrocytes, 1, a - increase in volume 10 × ok.10, 1, b - increase in volume 40 × ok.10);

in FIG. 2a, 2b, the thyroid gland of a rat with hypothyroidism (21 days), a decrease in the diameter of follicles, an increase in the area and height of the thyrocyte, pronounced extrafollicular proliferation of thyrocytes, 2, a - an increase in volume of 10 × ok.10; 2b - magnification about 40 × approx. 10.

in FIG. 3, a, 3, b - the thyroid gland of a rat with hypothyroidism who received dry white cinquefoil dry extract (21 days) at a dose of 50 mg / kg, 3, a - increase in volume 10 × ok.10, 3, b - increase in volume. 40 × approx. 10.

in FIG. 4, a, 4, b - the thyroid gland of a rat with hypothyroidism treated with kelp (21 days) at a dose of 3 mg / kg, 4, a - increase in volume 10 × approx. 10, 4, b - increase in volume 40 × ok .10.

in FIG. 5 - the effect of dry white cinquefoil extract and kelp on the processes of free radical oxidation of biomacromolecules in experimental hypothyroidism (21 days);

in FIG. 6 - the effect of the cinquefoil of dry white extract on the peripheral deiodination of thyroxine with liver damage CCl ₄ in rats;

in FIG. 7 is a micrograph of particles of white cinquefoil extract.

The proposed pharmaceutical composition is made and acts on the body as follows.

White cinquefoil (Potentilla alba L.) is a perennial herbaceous plant of the family Rosaceae (Rosaceae). The area of growth is the central regions of the European part of Russia, Crimea, Central Europe, the Balkan Peninsula [Flora of the USSR, Volume X, Moscow, 1941; D. Randushka, L. Shomshak, I. Gaberova, Color atlas of plants, Bratislava, 1990].

White cinquefoil is widely used in folk medicine. For medicinal purposes, all parts of the plant are used. Extracts from aboveground and underground organs stimulate the central nervous system, exhibit antibacterial activity, have an astringent, antiseptic, wound healing and hemostatic effect. Phytotherapists recommend the use of white cinquefoil for the prevention and treatment of diseases of the liver, cardiovascular system and gastrointestinal tract [V.G. Drobotko, B.E. Eisenman, M.O. Schweiger et al., Antimicrobial substances of higher plants, Kiev, 1958; Modern phytotherapy / Under. ed. V. Petkova, "Medicine and Physical Education", Sofia, 1982; N. Stoyanov, Invest in the medicine of a plant: at 2 p., P. 1, Sofia, 1972; E.F. Hemp, L.V. Nikolaychuk, L.A. Bazhenova, Medicinal plants, Minsk, 2000; V.A. Kayukova, People's Doctor. - 2004. - No. 16 (182) - p. eighteen].

Of particular interest is the use of the roots and rhizomes of the white cinquefoil for the treatment of thyroid diseases. There are reports of the successful use of extracts from this plant for Bazedov-Graves disease [G.K. Smik, V.V. Krivenko, Pharmaceutical journal, No. 2, 58-62, 1975; IN AND. Todorova, Abstract of the dissertation of the candidate of pharmaceutical sciences, Kiev, 1997; E.I. Prikhodko, Medical practice, No. 6, 87-89, 1976; A.V. Zakharia, Abstract of the dissertation of the candidate biologist, sciences, Lviv, 1997].

The beneficial properties of the white cinquefoil are due to its unique chemical composition; it has been established that the underground part contains carbohydrates (starch); iridoids; saponins; phenol carboxylic acids; tannins (gallotanin) [O.M. Gritsenko, G.K. Smik, Pharmaceutical journal (Ukraine), No. 1, 88, 1977; CM. Loos, Introduction and acclimatization of roslin in Ukraine, Kiev, no. 14, 101-104, 1979; E.T. Sluka, M.A. Dachishin, Abstracts of the 3rd Congress of Pharmacists of the Ukrainian SSR, Kharkov, 253-254, 1979].

// J. Ethnopharm. - 2009. - Vol. 122. - P. 184-204.; Free radical scavenging activity of extracts obtained from cultivated plants of Potentilla alba L. and Waldsteinia geoides L. / A Matkowski, [et al] // Herba polonica. - 2006. - Vol. 52, №4. - P. 44-46].When studying the chemical composition of the underground organs of white cinquefoil, phytosterols, terpenoids, saponins, phenolcarboxylic acids, flavonoids, hydrolyzable tannins and proanthocyanidins were found [Isolation of (+) - catechin from the roots and rhizomes of white cinquefoil / A.I. Meshkov et al. // V Russian Scientific and Practical Conference: Actual problems of nanobiotechnology and innovation with unconventional natural resources and the creation of functional products. - 2009, S. 101-103; Isolation and identification of phytosterols from the roots and rhizomes of the white cinquefoil / A.I. Meshkov et al. // Questions of biological, medical and pharmaceutical chemistry. - 2009, No. 2, p. 36-37; Isolation of phenolcarboxylic acids from the roots and rhizomes of a white cinquefoil / A.I. Meshkov et al. // Questions of biological, medical and pharmaceutical chemistry. - 2013, No. 5, C 39-40; Khisyamova D.M. Comparative pharmacognostic study of some representatives of the genus Cinquefoil (Potentilla L.) // Abstract. Cand. dis. - Samara. - 2017 .-- 25 s; Antioxidant tannins from Rosaceae plant roots // J. Oszmianski, A. Wojdylo, E. Lamer-Zarawska, K. Swiader // Food Chem. - 2007. - Vol. 100, N 2. - P. 579-583; Tomczyka, M. Potentilla - A review of its phytochemical and pharmacological profile / M. Tomczyka, KP

// J. Ethnopharm. - 2009. - Vol. 122. - P. 184-204 .; Free radical scavenging activity of extracts obtained from cultivated plants of Potentilla alba L. and Waldsteinia geoides L. / A Matkowski, [et al] // Herba polonica. - 2006. - Vol. 52, No. 4. - P. 44-46].

One of the modern areas of drug development is the production of tablets and capsules, which include dry extract of the roots and rhizomes of the white cinquefoil. Capsules and tablets have been developed containing finely divided powder of the white cinquefoil grass [BY 20101375]. Composition for the preparation of a medicament for the treatment and prevention of thyroid diseases). The disadvantage of this pharmaceutical composition is the low bioavailability of biologically active substances from capsules and tablets and their high microbial contamination, because medicinal plant material (white cinquefoil grass) is only dried and crushed in the absence of a stage for obtaining a dry or thick extract. To solve this problem, a technology has been developed to obtain a dry extract of the roots and rhizomes of white cinquefoil, which consists in the use of raw materials of three- or five-year-old cultivated plants, the degree of grinding 3-5 mm, triple extraction with 40% ethyl alcohol with heating 65-70 ° C and constant stirring , water module 1:15, followed by concentration of the extract and drying at a temperature not exceeding 60 ° C.

The dry extract of the roots and rhizomes of the white cinquefoil is a light-hygroscopic amorphous powder of light brown color with a weak specific odor and a strongly astringent taste. This is a standardized dry extract that meets all quality indicators for dry extracts.

As a result of studies of the micro- and macroelement composition, it was found that the underground mass of the plant is a concentrator of such elements as Al, I, B, Ba, Mn, Zn.

A phytochemical study showed the presence of the following groups of biologically active substances in a dry extract: phytosterols, triterpenoids, organic and phenolcarboxylic acids, mono and oligosugars, tannins (condensed type).

From the extract of the roots and rhizomes of the white cinquefoil, the following were identified and identified: (+) - catechin, (+) - epicatechin and their monoglycosides; p-coumaric and ellagic acids; β-sitosterol, β-sitosterol-3-O-β-D-glucopyranoside (daucosterol); tormentic acid (tormentol, tormentic acid); 5 monosugars (galactose, xylose, arabinose, glucose, rhamnose), more than 30 type A and B proanthocyanidins, as well as their glycosides.

The main compounds of the dry extract are condensed tannins, derivatives of catechin and epicatechin - proanthocyanidins. In terms of (+) - catechin, their content reaches more than 50%.

The pharmacological activity of the dry extract of white cinquefoil (SELB) is due to the amount of biologically active substances included in its composition.

The effect of the dry extract of white cinquefoil in various concentrations on the functional activity of platelets in vitro under the conditions of ADP - induced aggregation of rabbit platelets was studied.

To assess the effect of the studied extract on the platelet link of hemostasis, as one of the important pharmacological properties of the object, and to expand the understanding of the mechanism of action of the dry extract of white cinquefoil.

Research method.

The pharmacological activity of the cinquefoil extract was studied using venous blood of outbred male rabbits weighing 2.8-3.5 kg when taking from the marginal vein of the ear by free fall under stabilization with a 3.8% sodium citrate solution in a ratio of 9: 1. Platelet aggregation was studied by light scattering according to the method of G. Born, on an aggregometer manufactured by BIOLA (Russia). ADF (NPO RENAM, Russia) at a final concentration of 1 × 10 ^{-5 was} used as an aggregation inducer. The degree of aggregation was judged by the maximum optical density drop after the end of the reaction (A max) compared to the initial value. Statistical data processing was performed using t-student criterion.

Research results.

Potentilla extract was studied in concentrations of 0.01-10.0 mg / ml. As a comparison drug used aspirin in similar concentrations. The results of the study are presented in table. one.

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05.

It was shown that the studied extract in the concentration range of 0.01-10.0 mg / ml had a small but significantly inhibitory effect on platelet aggregation, reduced the degree of aggregation by 3-18% compared with the control, which indicates its anti-aggregation properties. In terms of anti-aggregation activity, the extract is significantly inferior to the activity of the comparison drug, aspirin (aspirin reduces the degree of platelet aggregation by 49-90%).

As a result of the experiments, it was found that the extract of white cinquefoil has insignificant anti-aggregation activity under conditions of ADP-induced aggregation.

2. The effect of the dry extract of the roots of the white cinquefoil (SELB) on aspirin gastric ulcers in rats was studied.

Rats weighing 155-235 g are left to starve for 24 hours, without limitation of water. After dividing the animals into groups (two experimental and one control), animals are injected with aspirin at a dose of 200 mg / kg (1% aqueous solution boiled and cooled). After 1 hour, rats of the experimental group were administered SELB at doses of 50 and 100 mg / kg. After another 1 hour they give food. This sequence of injections lasts 3 days. On the fourth day, rats are killed by decapitation, the stomachs are removed and a macroscopic examination of the mucosa is performed. The results are presented in table 2.

In this model of experimental gastric ulcers, a weak gastroprotective effect of SELB has a dose of 50 mg / kg. The extract quantitatively inhibits the development of ulcerative defects of the mucosa (Pauls index 2.9; therapeutic efficacy 2.1), without affecting their area (Pauls index 4.4). At a dose of 100 mg / kg, the gastroprotective effect of the cinquefoil is weakly expressed.

3. The study of antiexudative activity SELB.

The effect of the dry extract of the roots and rhizomes of the white cinquefoil on the course of formalin swelling of the paws in mice was studied. Edema was caused by introducing into the aponeurosis 1% formalin solution in a volume of 0.05 ml. White mice weighing 17-19 g were in the experiment. The action of the extract was studied at doses of 50 and 100 mg / kg when administered orally one hour before formalin and one hour after the administration of the agent. Control animals were injected with physiological saline in a similar volume. After 3 hours at the peak of inflammation, the animals were killed and the increase in foot volume was recorded by weighing the amputated inflamed and uninflamed paws of the animal on a VT-500 torsion balance. The experimental data processed by the method of variation statistics are presented in table 3.

As can be seen from table 3, the extract has a weak anti-inflammatory activity. The dose-dependent effect of the drug was noted. A clearer antiexudative effect was established at a dose of 100 mg / kg, compared with the control, the effect was 12.0%, at a dose of 50 mg / kg it was 3.0%, respectively.

4. The study of the effect of cinquefoil extract on the course of the chronic proliferative phase of inflammation in mice.

The effect of SELB on the course of the chronic proliferative phase of inflammation was studied according to the Meier method (1950). White outbred mice weighing 20-22 g were used in the experiment. Sterile cotton balls weighing 4 ± 1 mg were implanted under the skin of mice in the axillary and inguinal region. SECKLB was administered orally at doses of 50 and 100 mg / kg immediately after implantation and for the next 7 days. On the 8th day, granulomas were extracted, which were weighed first in the wet state, then in the dried state to a constant weight at a temperature of 80 ° C. The proliferative activity of the preparations was calculated by the difference between the mass of dry granuloma and the mass of a cotton ball (4 mg), and the antiexudative activity was calculated by the difference between the mass of wet and dry granulomas. The results processed by the method of variation statistics and are presented in table 4.

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05.

As can be seen from table 4, the extract at a dose of 50 mg / kg has antiproliferative activity, inhibits the formation of fibro-granulation tissue against chronic proliferative inflammation in mice by 26.7%. At a dose of 100 mg / kg SELB enhances the formation of fibrous-granulation tissue by 19%, respectively.

5. The study of the analgesic activity of the dry extract of white cinquefoil (SELB).

To study the effect of SECKLB on the threshold of pain sensitivity, we used the “cramp” model of pathological pain. In the experiment were white outbred mice weighing 22-24 g. Dry extract of the roots and rhizomes of the white cinquefoil was injected orally at a dose of 50 mg / kg. Control animals received physiological saline in the same volume. After 45-60 minutes, against the background of the extract, the “cramp” model was reproduced. The pain reaction was caused by chemical irritation, by intraperitoneal injection of 0.25 ml of a 0.75% solution of acetic acid. After that, the number of specific nonoceptive responses of the “cramp” type was recorded within 10 minutes.

The severity of analgesia, after administration of the drug was evaluated by the formula: A = (Pk-Ro) Pk × 100%, where A is the coefficient of analgesia; Pk - the number of “cramp” type responses in 10 minutes; Po - the number of "cramp" type responses in 10 minutes after drug administration. The experimental data are presented in the table.

Note * - P <0.05 compared with control

From table 5 it is seen that SELB at a dose of 50 mg / kg have analgesic activity. Under the action of the extract, the number of specific nonoceptive responses to a chemical stimulus decreased by 26.4%. The number of “cramps” was: 26.00 ± 3.09, while in the control group it was 35.33 ± 3.11, respectively.

As a result of the experiments, it was found that the cinquefoil of white roots and rhizomes, the dry extract at a dose of 50 mg / kg, has antiaggregatory, gastroprotective, anti-inflammatory and analgesic effects.

The thyroid-stimulating effect of the dry extract of the roots and rhizomes of the white cinquefoil has been established.

The effect of cinquefoil of dry white extract on the course of experimental hypothyroidism was studied.

The studies were performed on white Wistar rats of both sexes weighing 180 ± 12 g from January 10 to October 30, 2012.

In order to study the thyroid-stimulating properties of the cinquefoil of the dry white extract, a model of experimental hypothyroidism in white Wistar rats was reproduced. As a comparison drug, kelp was used (Evalar, Russia) at a dose of 3 mg / kg, which replenishes the daily norm of iodine (according to the annotation). Kelp is known to be a natural source of iodine. The iodine content is 2.7-3.0% and for the most part it is in the form of iodides and diiodotyrosines, which differ in pronounced bioavailability.

As a result of daily administration to rats of mercazolil at a dose of 10 mg / kg orally for 28 days, symptoms consistent with the clinical picture of hypothyroidism were observed. Rats showed hypodynamia and an increase in body weight by 45.0% compared with those in animals in the intact group. An enzyme-linked immunosorbent assay in the blood serum of rats showed a decrease in the level of thyroid hormones in comparison with the data of the intact group of animals: T ₄ - by 43%, T ₃ - by 15%, and the level of TSH increased by 6.0 times (Table 6). The physiological (normal) levels of thyroid hormones in rat serum were taken as those of the intact group of animals in this experiment. A decrease in the deiodation index in animals of the control group, which characterizes an increase in peripheral deiodination against the background of an increase in the level of TSH in the blood serum, may indicate the mobilization of the adaptive - compensatory reaction of the animal organism in conditions of iodine deficiency, aimed at the formation of the most active thyroid hormone.

Thus, the development of experimental hypothyroidism in white rats is indicated by the levels of thyroid hormones in the blood serum of animals, morphological signs of gland damage, the clinical course of gland dysfunction, behavioral reactions in animals, and other data.

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05. ELBS - dry white potentilla extract, n - number of animals in the group.

Against the background of course administration to animals per os of an aqueous solution of white cinquefoil extract for 21 days at doses of 10 and 50 mg / kg, the hormonal background in animals is restored, as evidenced by the concentration of hormones T ₃ , T ₄ , TSH in serum (Table. 6). The weight of the rats of the experimental groups is reduced slightly compared with the data in the control group of animals.

As can be seen from table 6, the course introduction to animals of the extract of white cinquefoil at a dose of 10 mg / kg on the 21st day of the experiment leads to an increase in the levels of the hormone T ₄ by 65.0%, the hormone T ₃ by 3.0%, while the level of TSH remains high in animals of this experimental group. Moreover, the correction of the levels of hormones T ₃ and TSH occurs more slowly than T ₄ . Probably, a longer course time of the indicated drug is needed than 21 days to restore the hormonal background with the extract at a dose of 10 mg / kg. Moreover, the deiodation index corresponded to that in the intact group of animals. The course administration of the studied extract to another experimental group of rats at a dose of 50 mg / kg at the end of the experiment led to an increase in T _{4 by} 2.9 times, T ₃ by 40% and a decrease in TSH level by 50.0% compared with the same values in the control group of animals. An increase in the deiodination index in animals of the second experimental group indicates the suppression of peripheral conversion of T ₄ century. T ₃ . The comparison drug had a pronounced effect on the production of hormones, increasing the level of T ₄ 2.4 times and decreasing TSH 2.2 times.

Thus, cinquefoil dry extract in the indicated doses increases the functional activity of the rat thyroid gland in the experiment. It was found that against the background of the introduction of white cinquefoil extract at a dose of 50 mg / kg, the level of TSH reaches optimal values, decreasing by half.

2. The effect of cinquefoil extract on the morphofunctional state of the thyroid gland under experimental hypothyroidism.

In the study of histological sections of the thyroid gland of the intact group of rats (Fig. 1, a, Fig. 1, b), a typical follicular structure of the gland was observed with hematoxylin and eosin staining, connective tissue septa separating lobules from each other were visible. Follicles are rounded or slightly oval in shape with clear contours. The colloid is pale pink. The cells of the follicular epithelium were mainly cubic in shape, the nuclei were located mainly at the basement membrane of thyrocytes.

On histological sections of the thyroid gland of the control group (Fig. 2, a, Fig. 2, b), receiving mercazolil for 28 days, a morphological picture inherent in iodine deficiency is noted. Due to the development of hypothyroidism of the thyroid gland, the thyroid-stimulating function of the pituitary gland increases; under the influence of an excess of TSH, the thyrocytes were in a state of functional activity. Follicular epithelial cells looked swollen, acquired a prismatic or cylindrical shape. Thyrocytes in some places in the apical membrane region had protrusions in the lumen of the follicles, in some zones the cells were in a state of mitosis, differentiation was noticeable, and it was clearly seen that 5-6 cells, grouped by a new basement membrane, began to synthesize and secrete thyroglobulin into the intercellular space. which in the form of a colloid filled it.

The presence of medium follicles) (130.5 ± 8.6 μm) with scalloped edges, high follicular epithelium (31.50 ± 12.31) and a small amount of colloid were also detected. There are many follicles (76.5 ± 5.4 μm) of small diameter (up to 100 μm). Due to the slowdown in utilization, the colloid became denser, became heterogeneous, contained many vacuoles, and its resorption from follicles was observed. In the gland, zones of increased functional activity of the follicles and areas where proliferative processes (cell clusters) predominated (Fig. 2a, Fig. 2b) predominated were simultaneously detected. The thyrocyte area in the control was 4.6 times larger than that of intact rats. So, the total volume of follicular epithelium significantly increased due to extrafollicular thyroid cells. The diameter of the follicles decreased by 26.0%, the amount of colloid decreased by 24.0%, the height of the thyrocyte increased by 47.0% compared with the indices of the intact group (Table 7). The increase in the number of mitoses was characteristic for the indicated observation period. Thus, the thyroid gland when exposed to mercazolil at a dose of 10 mg / kg for 28 days corresponds to morphological signs of the state of inhibition of its function. A similar picture of the development of experimental hypothyroidism is described by B.V. Aleshin (1987), S.A. Stepanov (1997), C.D. Zhamsaranov (2007). Such a picture against the background of iodine deficiency can be a prerequisite for nodule formation, which happens with endemic goiter.

On histological sections of the thyroid gland of the experimental group of rats treated with dry white bloodroot extracts at a dose of 50 mg / kg (Fig. 3, a, Fig. 3, b), a decrease in the activity of hyperplastic processes in the organ stroma was observed, the colloid had a lighter color, uniform consistency. The average follicle diameter increases by 42.0%, while the height of thyrocytes decreases by 47.0% compared with these indicators in the control group. At the same time, the volume of thyrocyte decreased by 65.0%, in contrast to the data of the control group of animals. Also, a decrease in proliferating thyrocytes in the stroma of an organ was found, follicles acquired a rounded regular or oval shape.

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05.

The follicular epithelium was approaching cubic, the average height of the thyrocyte decreased by 47.0% compared with the control. A 1.8-fold decrease in the activity of the mitotic index compared with the control indicated a decrease in extrafollicular proliferation of thyrocytes. A decrease in PKI characterized a decrease in thyroid activity, as evidenced by a decrease in proliferation of thyrocytes. The activity index of the thyroid gland and the Brown index on the 21st day after the course administration of the indicated agent were close to those of the intact group of rats. A similar pathomorphological picture was observed in the group of rats treated with

The effect of dry white cinquefoil extract on the state of the central nervous system of rats under experimental hypothyroidism.

It is known that iodine deficiency leads to deep disorders of higher nervous activity: impaired intelligence, changes in behavioral reactions; patients with hypothyroidism are predisposed to the development of depressive disorders and anxiety. At the same time, symptoms of mental depression come to the fore, appearing several years before the manifestation of an obvious clinical picture of hypothyroidism.

The possibility of the effects of experimental hypothyroidism and its correction by extract of dry white cinquefoil on the state of the nervous system in rats was studied in tests: “open field”, “elevated cruciform labyrinth”, conditioned passive avoidance reflex (passive avoidance reaction).

When studying the behavioral reactions of rats with experimental hypothyroidism, a decrease in overall motor activity was detected, which was judged by the horizontal and vertical activity of animals, compared with that in rats of the intact group. A decrease in the “mink” reflex (research activity) in rats of the control group was also noted. A decrease in motor activity with a decrease in research behavior in rats of the control group indicates a predominance of inhibitory processes in the nervous system. Against this background, a slight decrease in bowel movements and grooming was observed than in intact animals. The lack of research activity in combination with the acts of grooming suggests the stressful nature of animal behavior.

Observations of animals with experimental hypothyroidism described above indicate the predominance of inhibitory processes in the nervous system of rats, which may be a consequence of a decrease in the level of thyroid hormones in the blood of animals.

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05.

The course administration of white rats extract of white cinquefoil against white background at a dose of 50 mg / kg to white rats increases the horizontal and vertical activity by 8.0 and 3.0 times, respectively, compared with those in animals in the control group (Table 8). ) Activation of research activity in combination with a minor. a decrease in grooming acts indicates a more comfortable, more relaxed behavior of animals. A decrease in bowel movements by 24.0% compared with the data in the control group, combined with increased motor activity, allows us to conclude that there is a significant decrease in anxiety in rats of the experimental group treated with dry white cinquefoil extract. The comparison drug had a practically similar effect, but the total motor activity in the rats of the experimental group was 1.5-2.0 times higher than in the group of rats treated with kelp.

In the “elevated cruciform labyrinth” test, the level of anxiety of animals was studied. During the course administration of dry dry cinquefoil at a dose of 50 mg / kg to animals, a significant decrease in the time spent in the closed labyrinth arms by 14%, an increase in the time spent on rats at the central site by 3.7 times, as well as in open arms compared with the data, were recorded in the control group.

The residence time of the rats of the experimental groups in the open arms and on the central platform approached those of the intact group of animals. Rat peeping from closed sleeves and hanging downward was noted, indicating research activity and risk assessment in this situation. These indicators were similar for the comparison drug. Thus, against the background of administration of an extract of white dry cinquefoil at a dose of 50 mg / kg to animals, research activity is activated and the anxiety of rats during experimental hypothyroidism is reduced.

To assess the effect of dry white cinquefoil extract on the formation and preservation of a memorable trace, the passive avoidance conditioned reflex method (passive avoidance reaction) was used. In rats of the control group, passive avoidance reaction was not developed in our experiments. During the course administration of dry dry cinquefoil at a dose of 50 mg / kg to animals, antiamnestic functions were clearly expressed, in contrast to rats of the control group. The latent period increased by 1.7 times on the 1st day of observation, by 2.3 times on the 3rd day, and by 2.0 times on the 7th day compared with the same rats of the control group (Table 10).

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05.

The effect of cinquefoil dry white extract on the processes of free radical oxidation in experimental hypothyroidism.

As hypothyroidism progresses, a shift in the oxidative-antioxidant balance is observed towards lipid peroxidation. Thyroid hormones play a significant role in the mechanisms of the development of a stress reaction. Under stress, against the background of a decrease in adrenoreactivity, they have an antistress effect, so a deficiency of thyroid hormones can help increase stress sensitivity. In turn, disorders of redox processes exacerbate the severity of the course of hypothyroidism, forming a state of oxidative stress.

The intensity of the processes of free radical oxidation of biomacromolecules was evaluated by the accumulation of malondialdehyde (MDA) in blood serum. The state of the antioxidant system in rats was judged by the activity of superoxide dismutase (SOD) in blood erythrocytes, catalase and reduced glutathione in blood serum.

It was found that in experimental hypothyroidism in rats of the control group, there is an increase in the concentration of MDA by 37.0% and a decrease in the content of antioxidant defense enzymes: SOD - by 2.7 times and catalase - by 27.0% compared with data from the intact group of animals (Fig. . 5). These data confirm that in experimental hypothyroidism, the processes of free radical oxidation of biomacromolecules with the accumulation of lipid peroxidation products in the blood are enhanced.

The course administration of 50 mg / kg white cinquefoil to animals exerted a pronounced antioxidant effect, as evidenced by a decrease in the content of MDA by 18.0%, an increase in the activity of catalase and superoxide dismutase by 23.0% and 43.0%, respectively, and an increase in the content of reduced glutathione by 7.5% compared with similar indicators in animals in the control group. The antioxidant activity of the extract of white dry cinquefoil was slightly inferior to that of the comparison drug.

These studies have confirmed that with hypothyroidism, the processes of free radical oxidation of biomacromolecules are enhanced with the accumulation of products in the blood serum, in particular, MDA. In experimental hypothyroidism, changes in the state of the nervous system of animals occur, characterized by changes in behavioral reactions correlated with a description of human behavior in hypothyroidism - slowness, lethargy, anxiety, and depression.

A course administration of 50 mg / kg dry white cinquefoil extract to animals promotes an increase in the levels of thyroid hormones in rat blood serum, stimulating the morphofunctional activity of the thyroid gland in the experiment. The administration of the studied drug to rats also contributed to an increase in the orientation and research activity of white rats in a new environment, a decrease in the level of anxiety, and the preservation of a memorable trail in rats during experimental hypothyroidism amid mobilization of the endogenous antioxidant defense system of the body.

The effect of the cinquefoil of dry white extract on the peripheral deiodination of thyroxine in case of liver damage with carbon tetrachloride.

As you know, about 80% of the peripheral deiodination of thyroxine occurs in the liver. In this regard, it was of interest to determine the effect of cinquefoil of dry white extract on the functional state of the liver during intoxication of CCl ₄ . At 7, 14, 21 days from the start of the experiment, the level of triiodothyronine (T ₃ ) in the blood serum of the animals was determined by the radioimmunological method, the activity levels of membrane-bound enzymes of alanine aminotransferase (ALT), aspartate aminotransferase (ACT), the content of total bilirubin, β-lipoproteins were determined (LDL, HDL) and total protein in the blood serum of rats with toxic hepatitis.

An enzyme-linked immunosorbent assay of the content of thyroid hormones in the blood serum of the control group of rats with toxic hepatitis showed a decrease in the content of T ₃ on the 7th, 14th and 21st day of the experiment by 10.0%, 11.0% and 20.0%, respectively, compared with the data in intact group of animals, which characterizes against the background of intoxication with carbon tetrachloride a natural decrease in the content of T ₃ in the blood. This indicates the suppression of peripheral deiodination of thyroxine. Also, under the action of CCl ₄ , the activity of membrane-bound enzymes ACT and ALT increased 6.0 times on rats on day 7 from the start of the experiment, which indicates severe damage to liver cells (cytolytic syndrome), there was an increase in the total bilirubin content on days 7, 14 and 21 at 2.6; 2.1 willows 1.8 times, respectively, a decrease in lipoprotein fractions was observed, indicating a violation of the synthetic function of the liver (Table 11).

With the introduction of the extract of white cinquefoil at a dose of 50 mg / kg, T ₃ levels increased on 7, 14, 21 days by 4.4%, 23.0% and 20.0%, respectively (Fig. 6). The course administration of the cinquefoil of dry white extract at a dose of 50 mg / kg led to a decrease in the activity of ACT and ALT enzymes by 32.0% and 43.0%, respectively. A tendency towards a decrease in the total serum bilirubin in rat serum was also observed. With the course administration of dry dry cinquefoil to animals on the 7th day, an increase in the level of total protein by 35.0% is observed compared to the data in the control group. A similar pattern was observed on days 14 and 21 of the experiment.

To study the state of antioxidant protection in animals, the level of serum catalase and the concentration of MDA in the liver homogenate were determined. Liver damage with carbon tetrachloride caused activation of lipid peroxidation processes, as evidenced by an increase in MDA by 16.0% and a decrease in endogenous antioxidant defense, in particular, catalase activity by 31.0% compared with data in the intact group of animals (Table 11).

Note: * - hereinafter means that the difference is significant in comparison with the control at P≤0.05.

Thus, it was found that the white cinquefoil extract has pharmacotherapeutic efficacy in liver damage with carbon tetrachloride. With the course administration of the studied extract to animals, the signs of the severity of the cytolytic syndrome were reduced, metabolic processes in the liver of rats were normalized. There was a distinct tendency to increase the level of T ₃ on days 7, 14, and 21 from the start of the experiment against the background of the administration of white cinquefoil extract, which indicates the restoration of the processes of peripheral deiodination of thyroxine under the action of the studied agent. The mechanism of action of the dry white cinquefoil extract is obviously due to its ability to inhibit the processes of free radical oxidation of biomacromolecules and increase the activity of the endogenous antioxidant defense system of the body due to membrane-stabilizing activity established in the studied extract, in particular, hepatocyte and thyrocyte membranes.

To achieve the desired technical result, a pharmaceutical composition is proposed that contains as an active substance a dry extract of the roots and rhizomes of white cinquefoil and is made in the form of a solid dosage form - tablets, which contain microcrystalline cellulose (MCC-101) and lactose monohydrate (SuperTab as a filler) 24), as an anti-friction substance of calcium stearate, croscarmellose as a disintegrant in the following ratios of components (wt.%):

cinquefoil extract 42-43 microcrystalline cellulose 26-27 lactose monohydrate 24 croscarmellose 5 calcium stearate 1-2

The advantages of tablets as a dosage form of any drug are: dosing accuracy, protection of the active substance from exposure to light, air, moisture, elimination of its unpleasant taste and smell, high bioavailability, corrective ability, high aesthetics, the ability to set certain properties to medicines [Alekseev K .V., Kedik S.A., Blynskaya E.V., Lazareva E.E., Uvarov N.A., Alekseev V.K., Tikhonova N.V. Pharmaceutical technology. Solid dosage forms: study guide. / Edited by Professor S.A. Kedika. - M .: 2011. - 662 p.].

The maximum positive effect of the invention is achieved due to the correct combination of the components of the dosage form, the introduction of certain auxiliary substances, the sequence of their introduction.

When developing tablets, the technological and physicochemical characteristics of the extract were taken into account: moisture (4.8%), fractional composition (predominance of fractions from 50 to 120 microns), flowability (average), compressibility, particle size and shape.

Particles of the extract of dry white cinquefoil (Fig. 7) are plates of various shapes with uneven edges and a rough surface with a size of 20 to 150 microns.

To obtain a solid dosage form, the following were determined: humidity, flowability, fractional composition, bulk volume, angle of repose, porosity, compressibility.

The analysis was carried out in accordance with the requirements of GF XIII OFS 1.4.2.0016.15 “The degree of flowability of powders”, which contains methods for determining the following indicators: flowability, bulk volume, angle of repose.

In order to correct the physicochemical and technological characteristics of the dry extract and obtain a stable dosage form that meets the requirements of the Global Fund of XIII edition, the selection of excipients was carried out. Using anhydrous lactose, lactose monohydrate, MKTs-101, MKTs-102, potato starch, corn starch, Emdex and other excipients, tablet formulations of the compositions shown in Table 13 were prepared.

One of the tasks in the production of solid dosage forms is to achieve good flowability of the tablet mass, which is hindered by: friction between the contacting surfaces of the particles, preventing them from sliding; friction force between the surface and the wall of the loading funnel. To overcome the effects of friction, sliding substances are introduced into the tablet mass. These problems are especially serious on high-speed equipment. Magnesium, calcium, and zinc stearates are among the most effective moving agents. [Alekseev K.V., Kedik S.A., Blynskaya E.V., Lazareva E.E., Uvarov N.A., Alekseev V.K., Tikhonova N.V. Pharmaceutical technology. Solid dosage forms: study guide. / Edited by Professor S.A. Kedika. - M .: 2011. - 662 p.].

The data obtained show that calcium and magnesium stearates have some differences in moisture content and sorption capacity [Dubinskaya VA, Polyakov NA, Suponitsky Yu.L., Dementieva NN, Bykov VA The study of the moisture exchange of stearic acid, calcium stearate and magnesium stearate // Chemical and Pharmaceutical Journal. 2010.V. 44. No. 2. S. 41-45].

Calcium stearate was chosen as a moving substance, which provides a lubricating and sliding effect, increases the flowability of the tablet mixture and ensures uniformity of the dosage form [Khadzhieva ZD, Kuznetsov AV, Biryukova DV Technological aspects of the use of excipients in the production of drugs. Basic Research, 2012, No. 5-2, p. 436-440].

According to the results of studying the physicochemical and technological characteristics of the tablet masses, taking into account the compressibility of the tablet mass, and the strength of the tablets, it was found that the composition No. 1 has optimal characteristics, ensuring stability, the absence of changes in the appearance of the tablets during storage, and the average weight of the tablets.

The study of tablets with white cinquefoil extract according to such characteristics as tablet disintegration in accordance with the requirements of the State Pharmacopoeia of the XIII edition, OFS 1.4.1.0005.15, revealed the need to add croscarmellose sodium as a disintegrant to ensure the necessary Disintegration indicator (not more than 15 minutes) [ GF XIII edition, OFS. 1.4.2.0004.15 "Abrasion of tablets"].

Thus, lactose monohydrate and microcrystalline cellulose - 101 were chosen as fillers. Lactose has obvious advantages, such as low cost, high bioavailability, and MKTs-101 provides an optimal average tablet weight, tablet resistance to abrasion (not less than 97%) [ Alekseev K.V., Kedik S.A., Blynskaya E.V., Lazareva E.E., Uvarov N.A., Alekseev V.K., Tikhonova N.V. Pharmaceutical technology. Solid dosage forms: study guide. / Edited by Professor S.A. Kedika. - M .: 2011. - 662 p.].

The proposed ratio of active and auxiliary substances is found experimentally and provides stability and high bioavailability of biologically active substances.

It is known that one of the most affordable and economical methods in the manufacture of a new drug is the method of stepwise mixing of medicinal and auxiliary substances with subsequent analysis of the tablet mass. The absence of a granulation stage allows us to simplify and speed up the manufacturing process of tablets, making it more profitable from an economic point of view. In addition, the process of destruction of thermo- and moisture-sensitive compounds is excluded, thereby ensuring the stability of the obtained tablets.

Thus, the direct compression method is relevant for obtaining tablets with a hygroscopic dry extract of white cinquefoil.

A method of obtaining the proposed composition includes the following operations: sifting of components, including dry extract, sequential mixing of the extract with excipients, analysis of the obtained tablet mass, tabletting, dust removal, packaging. This process is optimal and eliminates the processes of wetting, drying and granulation, which significantly reduces the time to obtain the finished dosage form.

The proposed pharmaceutical composition in the form of tablets uses an active substance - white root roots and rhizomes dry extract - with their individual physicochemical properties and specific excipients in an original previously unknown combination of components for a tablet dosage form, which provides a solution to the problem and achieving a technical effect.

Conducted patent research did not reveal similar technical solutions, which allows us to conclude about the novelty and inventive step, because this technical solution does not follow explicitly from the current level of technology.

The proposed composition in the form of tablets expands the range of medicines for the prevention and treatment of thyroid diseases and is the first composition on the territory of the Russian Federation based on derivatives of plant origin, as well as the first composition proposed, which includes dry extract of the roots and rhizomes of white cinquefoil.

Thus, the proposal achieves the required technical result in expanding the arsenal of funds used for the prevention and treatment of thyroid diseases, increasing the effectiveness of prevention and treatment and in reducing the possibility of side effects in the treatment of thyroid diseases.

The domestic industry has all the means (materials, equipment) necessary for the manufacture of the proposed drug.

Claims (2)

A pharmaceutical composition for the prevention and treatment of thyroid hypothyroidism, made in the form of tablets and comprising white cinquefoil as a therapeutically effective active substance and target additives, characterized in that white cinquefoil as a therapeutically effective active substance is included in the form of a dry white cinquefoil extract made by grinding 3-5-year-old cultivated plants of roots and rhizomes with a degree of grinding 3-5 mm, triple extraction with 40% ethyl alcohol when heated 65-70 ° С and constant stirring, hydromodule 1:15, concentration, extraction and drying at a temperature not exceeding 60 ° С, and calcium stearate, croscarmellose, microcrystalline cellulose and lactose monohydrate were used as target additives in the following components, wt . %: cinquefoil extract 42-43 microcrystalline cellulose 26-27 lactose monohydrate 23-24 croscarmellose 5 calcium stearate 1-2

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