RU2502507C2 - Pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, action preventing gastropathies caused by nsaid, and method for preparing it - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, action preventing gastropathies caused by non-steroidal anti-inflammatory drugs. The above composition contains sodium diclofenac 5 - 25%, quercetin in the form of dihydrate or an anhydrous substance 10 - 40%, polyvinylpyrrolidone 10 - 50%, sodium lauryl sulphate 0.25 - 10%, microcrystalline cellulose 20 - 40%, croscarmellose sodium salt 1 - 15% and magnesium stearate 0.25 -5.0% at total weight of the composition. The invention also concerns the method for preparing the specified compound.

EFFECT: invention provides preparing the pharmaceutical composition with high anti-inflammatory, cardio- and chondroprotective activities.

2 cl, 2 dwg, 17 tbl, 16 ex

Description

The invention relates to pharmacy, as well as medicine, and relates to a pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, action against gastropathies caused by non-steroidal anti-inflammatory drugs (NSAIDs), and methods for its preparation, which can be used for the prevention and treatment of inflammatory and destructive diseases of various systems and organs.

It is known that NSAIDs in the pharmaceutical market of Ukraine is the largest niche of the pharmacotherapeutic group, covering more than 250 drugs in various dosage forms [1]. With the recognized therapeutic effectiveness of NSAIDs, their use has a number of limitations associated with frequent manifestations of side effects. For example, even short-term treatment with these drugs can lead to 25% of cases of side effects, and up to 5% - a serious threat to life [2].

Despite a significant assortment of NSAIDs, pharmacological and pharmaceutical science continues an intensive search for the creation of new, more effective and significantly safer drugs of this pharmacotherapeutic group. The goal in these studies is often the task of developing original substances with fundamentally new mechanisms of pharmacological action. Solutions to these problems are possible by searching for ways to modify the structure of the molecules of NSAID substances or by developing finished dosage forms with new pharmaceutical properties. Very promising is the creation of combined dosage forms of NSAIDs with other pharmacologically active substances [1].

The discovery of the main enzymatic factors of the directed pharmacological action of NSAIDs - two isoforms of cyclooxygenase (COX), namely COX-1 and COX-2, at one time, initiated the creation of a new generation of drugs - selective and specific COX-2 inhibitors [2, 3]. However, modern medical practice, for various reasons, still prefers well-known drugs of this pharmacotherapeutic group. Therefore, the most recognized NSAIDs are the dosage forms of diclofenac, indomethacin, ibuprofen and others, which are well studied for their therapeutic efficacy and side effects.

In order to expand the therapeutic capabilities of known NSAIDs and prevent side effects, it is important to find ways to modify their dosage forms. Diclofenac sodium (DN) preparations have found wide therapeutic application in medical practice in the treatment of many diseases, which, with significant therapeutic efficacy, unfortunately, have a number of side effects, primarily allergic reactions, and so-called NSAID gastropathies, can total up to 85% of cases. Even the short-term use of small doses of NSAIDs, including drugs, can lead to serious threatening health problems. In such cases, it is necessary to stop the use of NSAIDs and conduct corrective therapy. Dosage forms of DN were created in order to reduce the manifestations of NSAIDs-gastropathy, however, this did not solve the problem of their gastrotoxicity. The anti-inflammatory effect of DN is associated with inhibition of the synthesis of prostaglandins (GH) by blocking the key enzyme of the metabolism of arachidonic acid cyclooxygenase (COX). The isoforms of COX are known: COX-1, COX-2 and COX-3, performing a different regulatory role in the synthesis of GHGs. So, COX-1 controls the production of PG, which ensures the normal functioning of active cell molecules, platelet synthesis of thromboxane A2, endothelial cells and cells of the gastric mucosa, prostacyclin, PG E2, etc. COX-2 is involved in the production of PG, which are responsible for inflammation cell proliferation, cell destruction, mitogenesis, etc. In most cells, the basal level of COX-2 is quite low, however, the expression of the COX-2 gene can dramatically increase (more than 50 times) due to the development of inflammation Eden. The anti-inflammatory activity of NSAIDs is directly related to the inhibition of COX-2, while inhibition of COX-1 causes the development of side effects. The given generalized mechanism of action of all NSAIDs applies, in particular, to DNs, anti-inflammatory effects and side effects are described by the same components and fundamentally similar mechanisms. The well-known "classic brand" drugs of diclofenac are drugs with the trade name Voltaren® of the Novartis Pharma concern in various doses and in various dosage forms. [four]. Fast-acting drugs are Voltaren® rapid, sugar-coated tablets, 25 and 50 mg, No. 30, with the active substance diclofenac potassium. The rapid achievement of the drug with analgesia, reduction of inflammation and heat occurs due to the sufficiently high solubility of potassium salt, as it ensures the rapid creation of a sufficient therapeutic concentration of the active substance in the blood. With sufficiently high therapeutic indicators of the effectiveness of Voltaren® Rapid, caution is always given to possible cases of gastropathy, that is, occurrence of intestinal bleeding or development of ulcers in the gastrointestinal tract even if there is no history of peptic ulcers.

Also known is a pharmaceutical composition containing NSAIDs, a compound to enhance its pharmacological action, reduce toxicity and side effects, and target additives (Ukrainian Patent No. 79072, IPC :: A61K 31/35, A61K 31/616, A61P 29/00, 2007) . As agents that enhance the pharmacological effect of NSAIDs and reduce their toxicity and side effects, polyphenolic compounds of plant origin, in particular pure quercetin, or animal origin, a substance from the group of two basic carboxylic acids or amino acids or their salts were used. All of these substances are included in the pharmaceutical composition along with various NSAIDs.

However, the known composition does not provide high anti-inflammatory, cardio, and chondroprotective activity in combination with a minimal effect on NSAIDs-gastropathy.

A known method of obtaining a pharmaceutical composition containing diclofenac (RF patent No. 2424793, IPC A61K 9/70, A61K 31/195, A61P 29/00, 2011), consisting in the inclusion of MDs in a composite polymer film. DNs are preliminarily uniformly dispersed with a set of auxiliary substances, including hydroxypropyl methylcellulose, which functions as a food-grade polymer film. The pharmaceutical composition obtained by this method in the form of a chewing film, promotes the rapid release of MD in a dispersed form in the patient's oral cavity. The pharmacological and therapeutic possibilities of the pharmaceutical composition of the drug in the form of a food film are quite limited, since it is intended exclusively for convenient oral administration and does not solve the problem of NSAID gastropathy.

There is also known a method for the treatment of osteatrosis with the aim of preventing erosive and ulcerative complications of the stomach and duodenum caused by DN, using Quercetin Granules manufactured by SPC Borshchagovsky Chemical Farm as a protective agent (Ukrainian Patent No. 3903, IPC, 2009). The preparation of Quercetin Granules contains 40 mg of the active substance, quercetin, in 1 g of granules and it is prescribed in a dose of 1-2 g three times a day before meals in the same course as NAM additionally after its administration.

Well-known pharmaceutical compositions, methods for their preparation, unfortunately, do not provide high anti-inflammatory, cardio and chondroprotective effects, do not always show a significant effect on NSAIDs gastropathy.

The basis of the invention is the creation of such a pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, action against NSAIDs-gastropathy and the method of its production, in which with the introduction of new ingredients into the pharmaceutical composition, their specific quantitative composition, using new technological processes of obtaining, its high anti-inflammatory, cardio, and chondroprotective activity and maximum effect against NSAIDs-gastropathies would be ensured.

This object is achieved in that the pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, as well as with anti-NSAID gastropathies, contains NSAIDs diclofenac sodium and bioflavonoid quercetin, which enhances its pharmacological effect, reduces toxicity and side effects, as well as targeted additives, contributing to the formation of dosage forms. According to the invention, NSAIDs, an additional anti-inflammatory agent quercetin and as a solubilizer polyvinylpyrrolidone surfactant sodium laurisulfate are the main components of the pharmaceutical composition, when a certain technological operation due to mechanochemical interaction form a complex with the studied pharmaceutical and pharmacological properties.

In this case, the pharmaceutical composition contains DN in an amount of 5 to 25%.

In this case, quercetin in the form of a dihydrate or anhydrous substance is contained in an amount of from 10 to 40%.

In addition, as the target additives, the solubilizer polyvinylpyrrolidone in the amount of 10-50% and the surface-active agent sodium lauryl sulfate in the amount of 0.25-10% were selected.

Moreover, the fillers contain at least one binding agent, one baking powder and one lubricating agent.

Moreover, microcrystalline cellulose in the total amount of 10-85% was selected as a binding agent.

Moreover, croscarmellose sodium salt in the total amount of 1-15% was selected as a baking powder.

Moreover, magnesium stearate in the total amount of 0.25-5% was selected as a lubricant.

In addition, the pharmaceutical composition is in the dosage form of tablets coated with a polymer film or capsules.

The problem is also achieved by the fact that in the method for producing a pharmaceutical composition with anti-inflammatory, cardio and chondroprotective activity, as well as with anti-NSAID gastropathies, according to the invention, pre-DN, quercetin and target additives are mixed, compacted, ground, the resulting mixture is mixed with with pharmaceutically acceptable excipients, dry or wet granulation of the mixture is carried out, after which hard gelatin capsules are filled with granulate or pressed and the tablets are coated with polymer Lenka.

According to the invention, the developed method for producing a pharmaceutical composition allows to obtain dosage forms with a complex pharmaceutical composition in the form of coated tablets or capsules containing components in such quantitative limits, in%:

Diclofenac Sodium 5-25% Quercetin 10-40% Fillers up to 100%

Both active substances of the new drug were combined in it, taking into account the knowledge about their pharmacological and therapeutic properties obtained with the use of mono-preparations based on their substances. Selected target additives, due to their physicochemical properties, contribute to the solubilization of active substances and their sufficiently high dissolution in the aqueous physiological environment of the gastrointestinal tract.

The anti-inflammatory effect of quercetin has a somewhat unconventional mechanism, which is based on pronounced antileukotriene activity. However, it should be noted that the pharmacological effect of quercetin has many other positive properties, which allows us to predict its indispensability in the correction of the inflammatory process and the relevance in the composition of a composite drug with DN. The feasibility of using quercetin, taking into account its pharmacodynamics, in such a new drug is obvious. The mechanism of the anti-inflammatory action of quercetin is schematically reduced to the inhibition of 5-lipoxygenase (LOG-5), a key enzyme of the biotransformation of arachidonic acid after lipoxygenase metabolism. The biochemical transformation of arachidonic acid ends with the formation of end products - leukotrienes, which determine the occurrence of inflammatory processes. Thus, quercetin blocking the further stage of leukotriene synthesis provides inhibition of these negative processes. At the same time, leukotrienes are characterized by a decrease in the intensity of blood supply to organs, activation of lipid peroxidation (LPO) processes, etc. [5, 6]. Quercetin is characterized by angioprotective, vasodilator, anti-inflammatory, hepatoprotective, choleretic effect. It is able to reduce the intensity of the processes of free radical oxidation of lipids, proteinuria, hyperglycemia and also exhibit hypoazotemic and diuretic effects [7-9].

Quercetin, possessing an anti-leukotriene effect, prevents damage to the vascular endothelium of the ischemic site and reduces postischemic microcirculation disorders - pathological conditions caused by leukotrienes. Thus, quercetin protects the vascular endothelium not only from such aggression factors induced by ischemia, as LP, and from leukotrienes [10-12]. Quercetin, in contrast to traditional NSAIDs, not only does not exhibit ulcerogenic effects, but vice versa, creates a gastroprotective effect due to the ability to suppress the functioning of H + -K + -ATPase (decrease in the synthesis of hydrochloric acid). It is also able to stimulate the production of cytoprotective prostaglandin E2 by the cells of the gastric mucosa and reduce the number of reactive oxygen species in the tissue of the gastric wall [6].

The combined drug NAM with quercetin is unknown. The combination in one dosage form of two active substances with different physicochemical properties is a rather difficult task for pharmacy.

The solution of the problem in technical terms was carried out by the consistent development of the manufacturing technology of the pharmaceutical composition and in accordance with the pharmacopoeial methods of its research. First of all, the pharmaceutical development of the pharmaceutical composition began with an assessment of the physicochemical properties of DN and quercetin, primarily focusing on their ability to dissolve in aqueous media so that these active substances have maximum bioavailability when administered orally in physiological conditions of the stomach. Achieving the high solubility of both substances is possible only with the use of excipients with the properties of solubilizers and surfactants. Such solubility modulators are polyvinylpyrrolidone (povidone, GTV) and sodium lauryl sulfate (LSH), the quantitative values of their introduction into the pharmaceutical composition were determined during the pharmaceutical development of the composition, focusing on the solubility profiles of the samples, performing the solubility test. Studying the kinetics of dissolution of samples of the pharmaceutical composition, both granules for capsules and tablets, with variable mass values of PVHT and LSN, their optimal ratio with quercetin was established. It is known that, due to the crystal structure of quercetin, the solubility of its substance is very low (1 mg in 100 ml of water). By combining it with PVP by means of mechanochemical interaction, that is, by pressing (compacting) the initial granulate, the solubility of the bioflavonoid was increased to 200 mg in 100 ml of water. Thus, the resulting complex of quercetin with PVP is almost equally soluble in aqueous solutions with different pH values, which, after pharmaceutical requirements, mimic the physiological environment of the body. The results of the joint solubility of quercetin with various brands of PVP are shown in graph 1 (Fig. 1).

To ensure the physicochemical stability of the complex, the weight ratio between quercetin and PVP should be in the range 1: 1-1: 3. It should be noted that these mass values of PVP are also sufficient to increase the solubility of the substance of MD due to the formation of the corresponding complex compound. The povidones used in their molecular weights differ as follows: K90 - 1,000,000; K25 - 30000; C17 - 10000. Given the characteristics of the dissolution kinetics of the complex of quercetin with PVP, a sufficient amount of LSF, a surfactant, was selected that promotes the rapid and uniform wetting of the surface of the complex granulate. This also reflected in accelerating the dissolution of the complex in the initial periods of time and, thus, could create optimal conditions for the absorption by the walls of the stomach of the soluble form of quercetin. With significant hydrophobicity of the surface of DN crystals, these selected excipients, PVP and VLF, also contributed to the solubility of this substance, and thereby could increase the bioavailability of this NSAID. A positive effect on the solubility of quercetin also has a selected amount of PVP and VLF, as evidenced by the solubility profiles shown in graph 2 (figure 2).

All samples of the pharmaceutical composition, starting with a complex of active substances - quercetin and DN with PVP and LSN, were obtained under consecutive technological operations by dry or wet granulation of the ingredients of the complex and auxiliary substances. The prepared granules were used to fill the capsules or they were tabletted and the solubility of these dosage forms was examined in accordance with the Pharmacopoeia Dissolution test. The solubility characteristics of DNs and quercetin in the pharmaceutical composition indicated a significantly higher solubility of these active ingredients compared to the solubility of individual substances.

During the pharmaceutical development, a sample of a pharmaceutical composition of a certain composition was made (see Table 1), called by the authors “Diclocor” and found application in studies of pharmacological properties.

Table 1 Composition and ratio of ingredients in one capsule or tablet core of the pharmaceutical composition Diclocor No. p / p Name of ingredient in mg at % one 2 four 5 one Quercetin 40.00 20.00 2 Polyvinylpyrrolidone K-25 40.00 20.00 3 Diclofenac Sodium 25.00 12.50 four Microcrystalline cellulose 80.00 40.00 5 Croscarsmellose sodium salt 10.00 5.00 6 Sodium Lauryl Sulfate 3.00 1,50 6 Magnesium stearate 2.00 1.00 Content of capsule or tablet core: 200.00 100.00

The pharmaceutical composition of the given composition obtained in accordance with the dry granulation technology includes: mixing the active substances with excipients, compacting or briquetting the mixture, grinding, mixing the grinding with excipients and granulating, compressing the granulate into tablets or filling it with hard gelatin capsules. Wet granulation technology is characterized by a granulation stage of the mixture, where instead of a compactor or press, wet granulation equipment and a dryer are used, such as a mixer granulator with agitators, such as Rota P or a granulator-dryer of a fluidized bed, such as Hurtling.

We give specific examples of the invention. The following examples illustrate the main aspects of the present invention, but should not be construed as limiting.

Obtaining all samples of granules of the complex of active substances - quercetin and DN, with PVP was carried out under the conditions of granulation technological operations using the dry or wet method, using dry pressing (compaction) of a mixture of active ingredients, followed by grinding of the briquetted material. Wet granulation was performed in granulator-dryers in a vacuum or warm running air in devices with a pseudo-boiling layer. Capsules were filled into the obtained granules or they were tabletted, examining the solubility of these dosage forms according to the Dissolution test.

The solubility characteristics of DNs and quercetin in the pharmaceutical composition indicated a significantly higher solubility of the complex of active ingredients compared to the solubility of individual substances.

Thus, in the course of pharmaceutical development, a sample of the pharmaceutical composition was made, named by the authors of the invention “Diclocor”, which found, as a preparation of a certain composition, the use in research of toxic-pharmacological properties:

table 2 Composition and ratio of ingredients per capsule or tablet of the pharmaceutical composition Diclocor No. p / p Name of ingredients mg % one 2 3 four one. Quercetin polyvinylpyrrolidone 40.00 20.00 2. K-25 40.00 20.00 3 Diclofenac Sodium 25.00 12.50 four. Microcrystalline cellulose 80.00 40.00 5. Croscarsmellose sodium 10.00 5.00 6. Sodium Lauryl Sulfate 3.00 1,50 7. Magnesium stearate 2.00 1.00 8. Contents of capsule or tablet core 200.00 100.00

The pharmaceutical composition of the above composition obtained by dry granulation technology, including mixing the active substances with excipients, compacting or briquetting the mixture, grinding and mixing it with excipients, and finally granulate it is compressed into tablets or filled with hard gelatin capsules. Wet granulation technology is characterized by a granulation stage of the mixture, where instead of a compactor or press, wet granulation and drying equipment is used, such as a granulator-mixer with agitators, such as Rota P or a granulator-dryer, such as Huttling with a boiling layer.

The following embodiments illustrate aspects of the invention, but should not be construed as limiting.

Example 1. The substances quercetin, diclofenac sodium and polyvinylpyrrolidone (polyplasdone K-25) and sodium lauryl sulfate are mixed in a granulator and compressed using a compactor. Then, the resulting briquetted mass is ground in a mill, sieved and the previously prepared mixture of microcrystalline cellulose and croscarmellose sodium salt is added to the obtained granulate in a mixer. After 2-3 hours of mixing, magnesium stearate is added to the granulate and then stirring is continued for another 30 minutes. The finished granulate is transferred to fill gelatin capsules or tabletting. A sample pharmaceutical composition of this composition is shown in Table 2. In the case of tablets, the cores were coated with a Colorcon Opadry II film coating blend. The tests "Disintegration", "Dissolution" and "Dosage uniformity" were carried out in accordance with HFCs.

Table 3 Composition and ratio of ingredients of the pharmaceutical composition per capsule or tablet No. p / p Name of ingredients mg % one 2 3 four one. Quercetin 40.00 17.02 2. Polyvinylpyrrolidone K-25 80.00 34.04 3 Diclofenac Sodium 25.00 10.63 four. Microcrystalline cellulose 71.00 30.57 5. Croscarsmellose sodium 13.00 5.53 6. Sodium Lauryl Sulfate 3.00 1.27 7. Magnesium stearate 2.15 0.91 8. Contents of capsule or tablet core 235.00 100.00

Example 2. The substances quercetin, diclofenac and polyvinylpyrrolidone (polyplasdone K-25), and then sodium lauryl sulfate are placed in a granulator mixer type Rota P and stirred for 1.5-2 hours, microcrystalline cellulose and croscarmellose sodium salt are added. The mixture is stirred for 30 minutes, a sufficient amount of purified water is gradually added to it with stirring, the wet granulate is stirred for 2 hours at room temperature. Then granulate is dried by connecting vacuum and heating, for 3-4 hours. Dry granulate is dusted with magnesium stearate, stirring for 40 minutes and then transferred to the manufacture of finished dosage forms. Thus, the composition of the sample obtained by the pharmaceutical composition are shown in table 3.

Table 4 Composition and ratio of ingredients of the pharmaceutical composition per capsule or tablet No. p / p Name of ingredients mg % one 2 3 four one. Quercetin polyvinylpyrrolidone 40.00 13.33 2. K-25 140.00 46.67 3 Diclofenac Sodium 25.00 12.50 four. Microcrystalline cellulose 65.50 21.83 5. Croscarsmellose sodium 21.00 7.00 6. Sodium Lauryl Sulfate 5.00 1,67 7. Magnesium stearate 3,50 1.17 8. Contents of capsule or tablet core 300.00 100.00

Example 3. A mixture of quercetin, sodium diclofenac, polyvinylpyrrolidone (polyplasdone K-25) and sodium lauryl sulfate is transferred to a Hurtling type pseudo-boiling layer granulator-dryer, mixed for 30 minutes, then microcrystalline cellulose and croscarmellose are added with sodium salt, the purified air is stirred for 30 minutes. The granules are intensively stirred with vigorous stirring with a sufficient amount of purified water, the granules are mixed with heated air for an hour and then dusted with magnesium stearate for 30 minut.Takim way, a sample of the pharmaceutical composition is given in Table 4. The granulate was further used for other purposes.

Table 4 The composition of the sample pharmaceutical composition obtained in Example 3 No. p / p Name of ingredients mg % one 2 3 four one. Quercetin polyvinylpyrrolidone 40.00 16.67 2. K-25 40.00 16.67 3 Diclofenac Sodium 50.00 20.83 one 2 3 four four. Microcrystalline cellulose 92.00 38.33 5. Croscarsmellose sodium 15.00 6.25 6. Sodium Lauryl Sulfate 6.00 2,50 7. Magnesium stearate 2.00 0.83 8. Contents of capsule or tablet core 245.00 100.00

Example 4. The compacted granulate of the substances quercetin, diclofenac sodium, polyvinylpyrrolidone and sodium lauryl sulfate, obtained analogously to example 1, is placed in a granulator-dryer pseudo-boiling layer type Huttling. Sodium salt and microcrystalline cellulose and, at the end of magnesium stearate, are added to it with an interval of 30-40 minutes in succession with vigorous stirring of croscarmellose. The obtained granulate is transferred to the manufacture of capsules or tablets.

Table 5 The composition of the sample pharmaceutical composition obtained in Example 4 No. p / p Name of ingredients mg % one. Quercetin polyvinylpyrrolidone : 0.00 24.00 2. K-25 50.00 20.00 3 Diclofenac Sodium 25.00 10.00 four. Microcrystalline cellulose 92.00 36.80 5. Croscarsmellose sodium 16.00 6.40 6. Sodium Lauryl Sulfate 5.00 2.00 7. Magnesium stearate 2.00 0.80 8. Contents of capsule or tablet core 250.00 100.00

A preclinical study of the pharmaceutical composition was carried out using the Diclocor sample in the tests of the study in order to establish in full the toxicological and pharmacological properties of the future drug, which could become a promising drug.

Example 5. Studies of acute toxicity of Diclocor by oral administration.

Experimental studies were carried out according to the method of V.B. Prozorovsky [13] in the dose range of 500-1500 mg / kg by the amount of active substances, with the intragastric administration of Diclocor to mice and rats. Observations of animals were carried out within two weeks after administration of the drug. The results of the study are shown in table 1.

Table 5 Mortality rates in the study of acute Diclocor toxicity (n = 72) Dose mg / kg Lethal effect, death of animals / number of animals Average lethality,% the mouse rats the mouse rats 500 0/6 0/6 0 0 700 1/6 1/6 16.7 716, 900 2/6 3/6 33.3 50,0 1100 3/6 4/6 50,0 66.6 1300 4/6 5/6 66.7 83.3 1500 6/6 6/6 one hundred one hundred

The median lethal dose of Diclocor was calculated on the basis of the dependence of the lethality rate on the dose used by the probit analysis [13]. As a result of the calculations, it was determined that the LD _{50 of} Diclocor for mice is 1051.3 ± 52.5 mg / kg (404.2 mg / kg for DN), and for rats - 952.5 ± 45.2 mg / kg (366 5 mg / kg according to DN). According to the literature, the indicator of acute DN toxicity for mice when administered orally is 172–370 mg / kg [14–16], and for rats 53–95 mg / kg [17–19]. Therefore, the data obtained indicate a pronounced decrease in the toxicity of this NSAID when it is combined with quercetin in one dosage form. Thus, the results of studying the toxicological characteristics of Diclocor in mice and rats make it possible to classify this agent as a class of low toxic substances (501 mg / kg <LD ₅₀ > 5000 mg / kg), according to the generally accepted classification of toxicity according to K.K. Sidorov.

Example 6. The study of medium-effective doses of Diclocor in the development of experimental inflammation.

The study of medium-effective doses of Diclocor was carried out on a model of carrageenan edema of the extremities in rats [14], using the drug in doses of 5, 10 and 20 mg / kg in the amount of active substances, which corresponds to approximately 1/200, 1/100 and 1/50 of the LD index ₅₀ . The ED _{50 was} determined by the probit analysis of the activity-dose dependence as 3 hours after administration to the phlogogen [13]. The results of the study are shown in table 2.

Table 6 The effect of Diclocor on the course of carrageenan edema of the foot in rats 3 hours after administration to the phlogogen (n = 40) Experienced group Dose mg / kg The increase in foot volume, cu Antiexudative activity,% Control pathology 36.8- ± 2.1 - Diclocor 5,0 33.4 ± 1.6 9.2 ± 0.4 Diclocor 10.0 23.5 ± 0.9 * 36.1 ± 1.3 Diclocor 20,0 17.6 ± 0.7 * 52.2 ± 2.2 Note. * - P <0.05 relative to the control pathology group.

In the course of the calculations, Diclocor AU _{50 was} determined by antiexudative activity, which amounted to 18.24 ± 2.06 mg / kg by the sum of active substances, which corresponds to 7.02 ± 0.79 mg / kg by day.

Based on the results of a study of acute toxicity and moderately effective doses of Diclocor, the therapeutic index (TI) was calculated, which characterizes the breadth of the therapeutic effect of the drug (Table 3).

Table 7 Characterization of the breadth of the therapeutic effect of Diclocor A drug ED ₅₀ , mg / kg LD ₅₀ , mg / kg TI Relative TI by day Diclocor 18.2 (7.0 per day) 952.5 (366.5 per day) 523 4.4 DN 8.0 [14, 16] 95.0 [17] 11.9 one

The calculation results indicate a significantly greater breadth of the therapeutic effect of the studied drug in comparison with DN, which is due to both a decrease in toxicity and an increase in the activity of this NSAID, which, in turn, is due to the summation of the pharmacological effects of both substances and confirms the advisability of creating a combined dosage form on them basis.

Example 7. The study of the ulcerogenic properties of Diclocor by oral administration.

Preclinical study Diklokora ulcerogenic properties, according to the recommendations DEZ MoH [14] was carried out by oral administration of the drug to rats in a dosage range 18,2-145,6 mg / kg (amount of active ingredients), which corresponds to 1 IU ₅₀ - 8 ED ₅ about the drug, compared with DN at a dose of 48.0 mg / kg (average ulcerogenic dose (UD ₅₀ ) [16]. The level of Diclocor's ulcerogenic activity, depending on the dose, was calculated based on the median values in each group and the 50% level activity under the influence of Voltaren in a dose of UD50 (table 4).

Table 8 Ulcerogenic activity of Diclocor in rats (n = 60) Research team Ulcerogenic activity,% Diclocor 18.2 mg / kg 1.7 Diclocor 36.4 mg / kg 3.3 Diclocor 72.8 mg / kg 16.7 Diclocor 145.6 mg / kg 41.7 Voltaren 48.0 mg / kg 50,0

Then, the UD50 index was calculated using probit analysis [13] and tolerance index (IP) as the ratio of UD ₅₀ / ED ₅₀ , which indicates the safety of the drug from the digestive tract [20] (Table 5).

Table 9 Comparative indicators of the tolerance index of Diclocor and Nam A drug UD ₅₀ , MG / KG ED ₅₀ , mg / kg IP Relative IP per day Diclocor 156.2 (60.1 per day) 18.2 (7.0 per day) 8.6 1.4 DN 48.0 [16] 8.0 [14, 16] 6.0 one

The results of the studies allow us to conclude that there is a pronounced decrease in gastrotoxicity of DNs when combined with quercetin in the same dosage form. This confirms an increase in UD _{50 of} Diclocor by 1.3 times and IP by 1.4 times compared to NAM.

Example 8. The study of the antialterative activity of Diclocor on the model of scarified wounds in rats.

The anti-alterative properties of Diclocor were studied on a model of standard scarificated wounds in rats [14] compared with the monocomponents of the composition — quercetin and DN. The anti-alterative activity of the drugs was evaluated by their effect on the reduction of wound areas. The results of the study are shown in table 6.

Table 10 Anti-Alterative Activity of Diclocor,% (p = 40) Research drug Dose mg / kg 7 days 9 days 11 days 13 days 15 days Diclocor 18.2 32.75 ± 0.87 * 52.09 ± 1.39 * 58.06 ± 1.55 * 79.26 ± 2.11 * 100.0 Quercetin 11.2 16.52 ± 0.44 17.87 ± 0.48 27.96 ± 0.75 31.85 ± 0.85 48.86 ± 1.3 DN 7.0 10.72 ± 0.29 12.17 ± 0.32 15.05 ± 0.40 14.81 ± 0.4 4.55 ± 0.12 Note. * - P <0.05 relative to animals receiving reference drugs.

The results obtained indicate a significantly higher level of Diclocor activity in comparison with its monocomponents, since complete wound healing and, correspondingly, a one hundred percent level of anti-alterative action of the studied drug were recorded on the 15th day of the experiment. By the level of indicators of alternative activity, Diclocor significantly exceeded Quercetin and DN by 2.0-2.9 and 2.2-5.4 times, respectively. Thus, Diclocor has a positive effect on alternative inflammation in the model of scarification of wounds in rats, helping to accelerate the healing rate of wounds, reduce the area of wounds and exhibit significantly higher rates of alternative activity compared to quercetin and DN.

Example 9. An in-depth study of the antialterative properties of Diclocor on a model of furazolidone-isadrine myocarditis in rats.

The alternative properties of Diclocor were studied at a dose of 18.2 mg / kg on a model of furazolidone-isadrine myocarditis in rats [14], causing this pathology by the combined administration of 200 mg / kg of furazolidone and with an hourly interval of 40 mg / kg of isadrine, compared with the action DN in the equivalent dose. During the experiment, on the 5th day, the effect of Diclocor and the reference object on the functional state of the myocardium was evaluated according to the results of an ECG study (Table 7).

Table 11 The influence of Diclocor on the indicators of the functional state of rat myocardium with isadrine-furazolidone myocarditis (n = 40) Indicators Intact control Control pathology Diclocor, 18.2 mg / kg DN, 7.0 mg / kg Duration RR interval, s 0.140 ± 0.006 0.108 ± 0.004 * 0.127 ± 0.005 ** / • 0.112 ± 0.004 * PQ interval, s 0.046 ± 0.002 0.036 ± 0.001 * 0.042 ± 0.002 ** 0.040 ± 0.002 * QT interval, s 0.077 ± 0.003 0.056 ± 0.002 * 0.066 ± 0.003 * / ** 0.064 ± 0.003 * / ** QRS complex, with 0.024 ± 0.001 0.018 ± 0.001 * 0.023 ± 0.001 ** / • 0.019 ± 0.001 * Voltage P wave, mV 0.087 ± 0.003 0.061 ± 0.002 * 0.084 ± 0.003 ** / • 0.069 ± 0.003 * T wave, mV 0.158 ± 0.006 0.084 ± 0.003 * 0.109 ± 0.004 * / ** / • 0.097 ± 0.004 * / ** teeth R, mV 0.430 ± 0.017 0.320 ± 0.013 * 0390 ± 0.016 ** / * 0.339 ± 0.014 * Displacement ST from the contour, mm 0.20 ± 0.08 1.90 ± 0.12 * 0.65 ± 0.17 ** / • 1.40 ± 0.12 ** Notes: 1) * - p - ≤0.05 relative to intact animals; 2) ** - p - ≤0.05 relative to the control pathology group; 3) “- p - ≤0.05 relative to animals receiving NAM.

The data given in the table indicate that according to the intensity of the lesion and the destabilization of the functional state of the myocardium, the studied groups of animals can be placed as follows: control pathology → DN → Diclocor → intact control.

It was noted that, against the background of the use of Diclocor, normalization of the indicators of the myocardial functional state occurred (three-duration intervals RR, PQ, QT, QRS complex, voltage of teeth P, T, R), which had a probabilistic nature not only in relation to the control pathology group, but and animals that received NAM. Thus, under the conditions of the development of furazolidone-isadrine myocardial damage in rats, Diclocor has a significant cardioprotective effect, which is based on a decrease in the intensity of alternative processes, significantly exceeding the level of DN activity.

Example 10. The study of the antiexudative properties of Diclocor in models of aseptic inflammation caused by various phlogogenes.

A study of the antiexudative properties of Diclocor was performed on rat aseptic inflammation models caused by the following phlogogen: λ-carrageenin, histamine, serotonin, zymosan, and prostaglandin E2 [14]. After 3 hours after the reproduction of inflammation, the antiexudative activity of the study drug was evaluated by the degree of reduction of foot edema (relative to the control pathology group). Data on the antiexudative activity of Diclocor were compared with the efficacy of monocomponents of the combination. The results of the study are shown in table 8.

Table 12 Antiexudative activity (%) of Diclocor in conditions of the development of aseptic inflammation caused by various phlogogens (n = 200) Object of research. Dose mg / kg Phlogogen λ-carrageenin histamine serotonin zimozan prostaglandin E ₂ Diclocor 18.2 61.7 ± 2.7 * / • 32.3 ± 1.9 * / • 27.0 ± 1.6 * 24.4 ± 1.5 * / • 31.9 ± 1.9 * Quercet. 11.2 15.8 ± 0.4 17.9 ± 1.1 12.4 ± 0.7 19.6 ± 1.2 7.7 ± 0.5 DN 7.0 52.2 ± 1.7 21.8 ± 1.0 25.9 ± 1.2 10.1 ± 0.5 34.0 ± 1.6

Notes: 1) * - p <0.05 relative to animals treated with quercetin; 2) _ • - p <0.05 relative to animals that received NAM.

The obtained data indicate a high level of anti-inflammatory action of Diclocor in models of carrageenin and prostaglandin inflammation, due to the presence of DN in its composition with its anticyclooxygenase activity [14, 16]. At the same time, the effect on zymosan inflammation is realized due to the antileukotriene action of quercetin [21]. The activity of Diclocor on histamine and serotonin edema models is equally due to both of the above mechanisms. Thus, Diclocor has a complex mechanism of anti-inflammatory action, exerting an inhibitory effect on almost the entire known spectrum of inflammatory mediators, exceeding the level of activity of reference agents, which is associated with the synergistic effect of monocomponents on the anti-inflammatory properties of each other, and is realized by additive action.

Example 11. An in-depth study of the antiexudative properties of Diclocor on a model of toxic pulmonary edema in rats.

An in-depth study of the antiexudative properties of Diclocor was performed on a model of toxic pulmonary edema in rats caused by intraperitoneal administration of a 6% solution of ammonium chloride at a dose of 400 mg / kg [14]. During the study, the activity of Diclocor was evaluated by the dynamics of the mass coefficient of the lungs (MCL) compared with the activity of the monocomponents of the composition. The results of the study are presented in table 9.

Table 13 The influence of Diclocor on the course of toxic pulmonary edema in rats (n = 50) Study group Dose mg / kg μl,% Activity,% Intact control - 0.71 ± 0.03 - Control pathology - 1.52 ± 0.05 * 0 Diclocor 18.2 0.91 ± 0.03 * / ** / • / •• 75.4 ± 2.77 • / •• Quercetin 11.2 1.31 ± 0.05 * / ** / •• 25.3 ± 0.9 •• DN 7.0 1.02 ± 0.04 * / ** 61.7 ± 2.2 • Notes: 1) * - p 5≤0.05 relative to intact animals; 2) ** - p≤0.05 relative to the control pathology; 3) • - p≤0.05 relative to animals treated with quercetin; 4) •• - p≤0.05 relative to animals receiving NAM.

Analysis of the results of the study shows that under the influence of Diclocor, there is a significant decrease in HKB of animals and a corresponding increase in antiexudative activity not only in comparison with the control pathology group, but also with the action of the monocomponents of the composition. The activity in the group of animals treated with Diclocor exceeded the indicators for the group where Quercetin was used, 3.0 times and DN 1.2 times. Thus, a positive therapeutic and prophylactic effect of Diclocor on the course of toxic pulmonary edema was established, which was confirmed by high rates of antiexudative activity of this drug.

Example 12. The study of the antiproliferative properties of Diclocor on the model of "cotton" granuloma in rats.

Diclocor's antiproliferative activity was studied using a “cotton” granuloma model in rats [14] compared with the action of monocomponents of the studied agent. The results of the study are presented in table 10.

Table 14 Diclocor antiproliferative activity in rat cotton granuloma model (n = 40) Study group Dose mg / kg The amount of granulation tissue, mg Activity,% Control pathology - 45.2 ± 1.7 - Diclocor 18.2 34.3 ± 1.3 * / • 24.12 ± 0.88 ** / • Quercetin 11.2 37.0 ± 1.4 * 18.14 ± 0.66 DN 7.0 39.1 ± 1.4 * 13.50 ± 0.49 Notes: 1) * - p≤0.05 relative to the control pathology; 2) ** - p≤0.05 relative to animals treated with quercetin; 3) • - p≤0.05 relative to animals receiving NAM.

The data of the studies showed that in terms of the degree of antiproliferative activity, Diclocor significantly exceeds the action of quercetin by 1.3 times and DN by 1.8 times. Thus, under the conditions of the development of proliferative inflammation, Diclocor has a pronounced antiproliferative effect, which is very useful in treating inflammatory and destructive diseases of parenchymal organs such as the liver, kidneys, heart, etc., since it can help suppress the development of sclerotic processes in them. .

Example 13. An in-depth study of the antiproliferative properties of Diclocoru on a model of doxorubicin cardiomyopathy in rats.

An in-depth study of the antiproliferative properties of Diclocor was performed under conditions of the development of doxorubicin-induced plastic myocardial insufficiency in rats, reproducing it by intraperitoneal administration of doxorubicin at a dose of 20 mg / kg [14]. During the experiment, on the 10th day, the effect of Diclocor on some biochemical parameters of laboratory animals was evaluated in comparison with the activity of DNs in an equivalent dose (Table 11).

Table 15 The effect of Diclocor on some biochemical parameters of rats with doxorubicin cardiomyopathy (n = 31) Research team Blood ASAT, mmol / year * L LDH blood, mmol / year * L TBA blood reactants, µmol / L Myocardial TBA Reactants, μmol / g Intact control (n = 12) 0.67 ± 0.03 5.13 ± 0.23 2.28 ± 0.07 63.62 ± 1.88 Control pathology (n = 4) 1.53 ± 0.14 * 7.77 ± 0.72 * 4.21 ± 0.24 * 151.61 ± 8.62 * Diclocor 18.2 mg / kg (n = 9) 0.81 ± 0.04 * / ** / * 5.98 ± 0.31 * / ** 3.55 ± 0.13 * / ** 97.23 ± 3.44 * / ** / • DN 7.0 mg / kg (n = 6) 1.33 ± 0.08 * 6.85 ± 0.43 * 4.02 ± 0.20 * 136.63 ± 6.74 * Notes: 1) * - P≤0.05 relative to intact animals; 2) ** - p≤0.05 relative to the control pathology group; 3) • - p≤0.05 relative to animals receiving NAM.

The results of biochemical studies showed an inhibitory effect on the part of the studied drugs on the accumulation of lipid peroxidation products and the growth of fermentemia in the development of cardiomyopathy. The most significant therapeutic effect was shown by Diclocor, which is confirmed by the likely dynamics of indicators relative to the control pathology. Moreover, the activity of AsAT and the content of TBA reactants in the myocardial tissue was significantly lower than in animals treated with DNs - 1.7 and 1.4 times, respectively. Thus, the data obtained indicate that Diclocor has, in addition to anti-inflammatory, also cardioprotective properties, which makes it expedient to use it in the treatment of inflammatory and destructive myocardial diseases.

Example 14. The study of the influence of Diclocor on the course of experimental autoimmune arthritis.

The study was conducted on a model of collagen-induced arthritis in rats, which was reproduced by subcutaneous twofold administration (with an interval of a week) of an emulsified mixture of bovine collagen type II and Freund's complete adjuvant (1: 1) at a dose of 2 mg / kg for collagen [22]. During the study, in the blood serum of animals as of 28 days, the dynamics of the content of necosanoids (prostaglandin E2 (PGE2), 6-keto-prostaglandin F1α (6-keto-PGF 1α), thromboxane B2 (TxB2) and leukotriene B4 (LTB4) was evaluated using enzyme-linked immunosorbent assay kits manufactured by Neogen Corporation (USA). The efficacy of Diclocor was studied in comparison with the action of its monocomponents in eclentic doses. The results of the study are shown in table 12.

Table 16 The effect of Diclocor on the content of eicosanoids in the blood serum of rats with collagen-induced arthritis (n = 50) Research team PGE ₂ , pg / ml 6-keto-PGF _1α , pg / ml TxB ₂ , pg / ml LTB ₄ , pg / ml Intact control 879.8 ± 12.5 261.6 ± 4.2 242.6 ± 9.5 261.4 ± 6.3 Control pathology 1248.4 ± 26.1 * 294.3 ± 4.1 * 493.2 ± 4.8 * 421.3 ± 4.7 * Diclocor 18.2 mg / kg 901.5 ± 17.5 ** / • 272.5 ± 6.7 ** 256.6 ± 8.9 ** / ' 345.6 ± 3.1 * / ** / • / •• Quercetin 11.2 mg / kg 1198.7 ± 22.1 * 286.8 ± 7.4 * 472.7 ± 6.7 * / ** 282.9 ± 8.4 ** DN 7.0 mg / kg 925.5 ± 13.6 * / ** 279.2 ± 5.4 * / ** 265.4 ± 5.9 ** 398.3 ± 6.5 * / ** Notes: 1) * - p≤0.05 relative to intact animals; 2) ** - p≤0.05 relative to control animals; 3) • - p≤0.05 relative to animals treated with quercetin; 4) •• - p≤0.05 relative to animals receiving NAM.

The study found that the use of Diclocor against the background of the development of collagen-induced arthritis contributes to the probabilistic normalization of the content of eicosanoids in rat serum. So, the level of PGE2 decreased by 1.4 times; the content of 6-keto-PGF1α - 1.1 times; TxB2 - 1.9 times and the level of LTB4 - 1.2 times. Moreover, in terms of the degree of pharmacological effect on most of the studied parameters, Diclocor exceeded the comparison drugs Quercetin and DN. The results obtained indicate that the drug Diclocor has a complex mechanism of action with an effect on both the cyclooxygenase (due to the content of DN) and lipoxygenase (the presence of quercetin) pathways of arachidonic acid metabolism. In this case, Diclocor is a promising corrector of inflammatory and destructive joint diseases with an autoimmune component, such as rheumatoid arthritis.

Example 15. The study of the influence of Diclocor on the structure of articular cartilage in the development of experimental osteoarthritis.

The effect of Diclocor on the morphostructure of articular tissues on the background of the development of experimental osteoarthritis was studied in comparison with the action of its monocomponents in equivalent doses. The studies were performed on a model of systemic steroid arthrosis in rats [23], which was reproduced by three times intramuscular injection of dexamethasone at a dose of 7 mg / kg with an interval of one week [24]. The test drugs were administered daily for 1 month, starting 2 weeks after the last injection of dexamethasone. During the morphological study of micropreparations of articular cartilage as of the 56th day of the experiment, morphometric determinations of its thickness, chondrocyte density and semi-quantitative assessment of the state of its main structures were performed (Table 13).

Table 13 Morphometric indicators of the articular cartilage of rats with experimental osteoarthrosis under the influence of Diclocor (n = 50) Study group Cartilage Thickness units Cell density according to the condition. units square The amount of points on the system for assessing the condition of cartilage Intact control 15.33 ± 1.15 42.40 ± 5.64 24.46 ± 0.22 Control pathology 12.62 ± 0.58 * 28.63 ± 1.85 * 12.68 ± 0.85 * Diclocor 18.2 mg / kg 15.73 ± 0.59 ** / * 35.83 ± 4.31 * / ** 20.19 ± 0.93 * / ** / * Quercetin 11.2 mg / kg 15.66 ± 1.35 32.67 ± 2.48 * 18.09 ± 0.94 * / ** / • DN 7.0 mg / kg 13.77 ± 0.52 32.20 ± 3.02 * 14.66 ± 0.95 *

Notes: 1) * - p≤5.05 for relatively intact animals; 2) ** - p≤0.05 relative to the control pathology group; 3) • - p≤0.05 relative to animals receiving NAM.

The results of the studies indicate that under the conditions of the development of experimental osteoarthritis under the influence of Diclocor, there is a significant increase in the thickness of the articular cartilage and the density of the location of chondrocytes on average by 20%, as well as the total score by a semi-quantitative system for assessing the state of cartilage tissue by 1.6 times. It should be noted that for most of these parameters, Diclocor significantly exceeded the activity of DNs and without significant differences quercetin. All of the above indicates that Diclocor has certain chondroprotective properties, determined by the content of quercetin in its composition. The use of this flavonoid as part of Diclocor allows you to positively modify the effect of DN on cartilage and significantly expand the possibilities of its use against the background of the development of degenerative-dystrophic lesions of the joints.

Example 16. The study of the processes of apoptosis under the influence of Diclocor in the development of experimental osteoarthritis.

Studies of the anti-apoptotic properties of Diclocor were carried out analogously to example 11. The identification of apoptotic cells in the tissues of the articular cartilage was carried out immunohistochemically on semi-thin paraffin sections on the 56th day of the experiment using the TUNEL reaction using the "In Situ Cell Death Detection Kit, AP" kits manufactured by "Roche Diagnostics" (Germany ) [25, 26]. In the course of the study, according to the results of the TUNEL reaction under the influence of Diclocor and its monocomponents in equivalent doses, a generalized quantitative assessment of the morphometric state of articular cartilage was performed (Table 14).

Table 17 Morphometric characteristics of the articular cartilage of rats according to the results of the TUNEL reaction (n = 50) Study group TUNEL- positive cells,% Intact control 1.1 ± 1.0 Control pathology 61.8 ± 5.8 * Diclocor 18.2 mg / kg 11.8 ± 3.7 * / ** / • Quercetin 11.2 mg / kg 16.3 ± 4.0 * / ** / • DN 7.0 mg / kg 40.7 ± 5.7 * / ** Notes: 1) * - p≤0.05 relative to intact animals; 2) ** - p≤0.05 relative to the control pathology group; 3) • - p≤0.05 relative to animals receiving NAM.

The data presented in the table indicate that under the influence of Diclocor against the background of developed osteoarthritis in rats, a marked decrease (5.2 times) in the intensity of apoptosis in cartilage tissue occurred. It had an unreliable level for quercetin and was significantly superior in DN activity. Thus, the basis of the mechanism of chondroprotective action of Diclocor is a positive negative effect on the prevention of chondrocyte apoptosis, which is reflected in a decrease in the number of TUNEL-positive cells. The results obtained make it possible to consider Diclocor a promising tool in the treatment of degenerative joint diseases, such as osteoarthritis, especially of secondary etiology.

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Claims (2)

1. A pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, action against gastropathy caused by non-steroidal anti-inflammatory drugs (NSAIDs), containing NSAIDs, a compound for enhancing its pharmacological effect and correction of side effects and targeted additives, characterized in that the selected NSAIDs diclofenac sodium in an amount of 5-25%, quercetin in the form of dihydrate with or without dihydrate was chosen as a compound to enhance its pharmacological action and correct side effects water substance in an amount of 10-40%, and polyvinylpyrrolidone in an amount of 10-50% and sodium lauryl sulfate in an amount of 0.25-10%, microcrystalline cellulose in an amount of 20-40%, croscarmellose sodium salt in an amount of 1- 15% and magnesium stearate in an amount of 0.25-5.0% based on the total weight of the composition. 2. A method of obtaining a pharmaceutical composition with anti-inflammatory, cardio- and chondroprotective activity, anti-gastropathy caused by non-steroidal anti-inflammatory drugs (NSAIDs) according to claim 1, characterized in that the formation of the pharmaceutical composition is carried out by dry granulation, which includes the following stages: mixing the active substances with fillers, compacting or briquetting the mixture, grinding the resulting briquettes, mixing grinding with fillers and granulation, extruded Ie granulate in tablets or filling them with hard gelatin capsules, followed by coating the tablets with a polymer film; or by the wet granulation method, which includes the steps of: mixing the active substances with fillers, moistening the resulting mixture, mixing the wet granulate with subsequent drying, dusting the granulate, compressing the granulate into tablets or filling hard gelatin capsules with it followed by coating the tablets with a polymer film.

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