EA039612B1 - Transdermal preparation for treatment and prevention of diseases of joints and soft tissues - Google Patents

Abstract

The invention relates to medicine and chemical-pharmaceutical industry, namely to transdermal agents for treating and preventing joint and soft tissue diseases, containing chondroprotectors and non-steroid anti-inflammatory agents as active components. Provided is a novel excipient formulation providing higher stability of glucosamine hydrochloride and ketoprofen while forming minimum amount of admixtures during storage. Storage life of the preparation with the provided formulation is at least 2 years. Higher stability of the active agent is achieved by using polyethylene glycol 400 and sodium metabisulphite in the formulation. Furthermore, the novel formulation preparation has an effective anti-inflammatory and analgesic action at ketoprofen concentration of less than 20% compared to commercial preparations, that allows to reduce the risk of developing an ulcerogenic effect from using this non-steroid anti-inflammatory agent.

Description

The invention relates to the field of medicine and the chemical and pharmaceutical industry, namely to transdermal preparations for the prevention and treatment of diseases of the joints and soft tissues, containing chondroprotectors and non-steroidal anti-inflammatory drugs as active components.

Osteoarthritis (hereinafter - OA) is a chronic progressive disease of synovial joints, the etiology of which is unknown. OA is considered to be a serious ailment of mankind, and the problem of treating this disease is one of the key problems in therapy, orthopedics, rheumatology, and clinical pharmacy. OA is widespread among elderly and mature people, characterized by degeneration of articular cartilage, structural changes in the subchondral bone, moderately or severely occurring synovitis, chronic course and a tendency to progression. All this leads to a decrease in working capacity (on average in 60% of cases) and leads to disability in patients of the most able-bodied age in 11.5% of cases [1].

This problem is of particular importance in connection with the increase in human life expectancy. According to epidemiological studies, dystrophic changes in the joints are found in 50% of cases in people over 40 years old, and at the age of 70 years, this disease was observed in 90% of the population. In general, the proportion of OA in the overall structure of the incidence of the population is 12% and ranks first among joint pathologies [1].

In OA, disorders are determined in all structural components of the joint, but the most pronounced changes are observed in the articular cartilage, according to which four stages of OA are distinguished. There are different points of view on histochemical and metabolic disorders in the articular cartilage in OA, in particular, a decrease in the content of glucosaminoglycans (GAGs) associated with the extinction of biochemical processes in chondrocytes. In addition, there is a decrease in the ability of cartilage to retain GAG macromolecules newly synthesized by chondrocytes in the matrix due to their transformation into a form that is not able to bind with hyaluronic acid. With the development of dystrophic and destructive disorders in the articular cartilage, the density of chondrocytes decreases due to their death [1,2].

The destruction of articular cartilage is possible due to the action of enzymes that are formed during inflammation. An essential link in the pathogenesis of the disease may be an increase in the biosynthesis of prostaglandins and their accumulation in the synovial fluid, which contributes to cartilage tissue damage and the induction of synovitis (inflammation of the synovial membrane). The inflammatory process in OA leads to the destruction of articular cartilage [1, 2].

OA is accompanied by pain, which increases with the progression of the disease [3]. Non-steroidal anti-inflammatory drugs (hereinafter referred to as NSAIDs) are used for systemic and local treatment of OA in acute and chronic pain [1-4]. The main mechanism of action of NSAIDs is the ability to inhibit the activity of cyclooxygenase (COX), thereby reducing the biosynthesis of prostaglandins and other inflammatory mediators. COX-1 is responsible for the synthesis of thromboxane and prostaglandins, which regulate physiological functions, in particular the protection of mucous membranes, and COX-2 is mainly responsible for the synthesis of prostaglandins, which are involved in the development of the inflammatory process and pain [4]. Inhibition of COX-1 leads to the development of erosive and ulcerative processes in the gastroduodenal zone [4]. This specific syndrome is called NSAID gastroduodenopathy. The mechanism of mucosal damage when using NSAIDs is as follows: inhibition of prostaglandin synthesis in the mucosa reduces the production of protective mucus and bicarbonates mediated by prostaglandins, which leads to the appearance of erosions and ulcers, which can be complicated by bleeding or perforation. Among NSAIDs, non-selective COX inhibitors have the most pronounced ulcerogenic effect, which, in particular, include indomethacin, ketorolac, diclofenac, piroxicam, and ketoprofen, which at the same time have a strong anti-inflammatory and analgesic effect [4]. Therefore, in OA, it is promising to use cutaneous drugs with non-selective NSAIDs, which ensures their transdermal penetration to the inflamed joint and an effective therapeutic effect with low bioavailability, which can reduce the frequency and severity of side effects [5-8]. At the same time, ketoprofen, which does not have a chondrotoxic effect, is a promising non-selective NSAID for the symptomatic treatment of OA [1].

Known drug Fastum® gel 2.5% containing ketoprofen at a concentration of 2.5% (information taken from the IMP, published 12/21/2010). This drug is most widely used for cutaneous application in the treatment of OA [1, 2, 7, 8]. However, this drug has a disadvantage; it is intended only for symptomatic therapy associated with the reduction of inflammation and pain, and does not contribute to the restoration of cartilage tissue.

Known preparations in the form of gels and creams containing chondroprotectors, such as Chondroxide® gel 5% containing 5% chondroitin sulfate. However, this drug has disadvantages associated with the fact that chondroitin sulfate has a large molecular weight of 20,000-30,000 a.m.u., as a result of which it cannot penetrate the skin and have a chondroprotective effect in OA, since optimal permeability is associated with a small molecular weight less than 500 amu [9-11]. Known drug Chondroxide® Maximum cream 8%, containing 8% glucosamine sulfate potassium chloride, having a low molecular weight, and used for the treatment of OA. However, this drug also has disadvantages; it is intended to provide only a chondroprotective effect and, due to the absence of NSAIDs in its composition, does not have anti-inflammatory and analgesic activity. The composition of the transdermal preparation does not contain penetration enhancers, which the developers tried to compensate for with a high content of the chondroprotector.

At the moment, patent RU 2582278 (CJSC PharmFirma Sotex, 04/25/2013) is valid in the Russian Federation, protecting a transdermal agent for the treatment and prevention of diseases of the joints and soft tissues, containing glucosamine or its pharmaceutically acceptable salt, ketoprofen, N-methylpyrrolidone (penetration enhancer) , propylene glycol, chondroprotective antioxidant (lidocaine and citric acid), acid gelling polymer and emulsifiers. This patent was chosen as a prototype, since the agent described in it contains a combination of the chondroprotector glucosamine and the NSAID ketoprofen, which are in a pharmaceutically acceptable base in a dissolved state, and the base provides their transdermal transport, thanks to N-methylpyrrolidone, with the manifestation of chondroprotective, anti-inflammatory and analgesic effects in experiments in vivo.

However, the tool described in the prototype does not meet the existing requirements for stability. Its shelf life is only about 1 year. As studies of the applicants have shown, under conditions of longer storage, the amount of formed decomposition products of ketoprofen and glucosamine exceeds the permissible limits, and the content of ketoprofen and glucosamine, respectively, decreases below the permissible limits, which does not allow mass production of this drug as a finished drug and sharply limits the possibility of its use in patients with OA only as an extemporaneous drug.

The objective of the present invention is to increase the stability and increase the shelf life of a drug containing ketoprofen and glucosamine in the form of a gel.

When creating such a composition, it is necessary to choose the excipients and the type of base in a balanced way, taking into account the fact that ketoprofen and glucosamine hydrochloride have different chemical and physicochemical properties. Ketoprofen is an organic acid, and glucosamine hydrochloride is a salt of an organic base. Glucosamine hydrochloride is readily soluble in water, while ketoprofen is readily soluble in ethanol and practically insoluble in water. Lowering the temperature during storage of the drug should prevent the decomposition of glucosamine, but crystallization of the dissolved ketoprofen may occur.

The problem is solved by creating a new composition of a transdermal agent for the treatment and prevention of diseases of the joints and soft tissues, containing the pharmaceutically required amount of a chondroprotector, a non-steroidal anti-inflammatory agent, an antioxidant for a chondroprotector, a gelling agent capable of forming gels in an acidic environment, mixtures of emulsifiers of the 1st and 2nd kind and mixtures solvents, where it contains glucosamine or its pharmaceutically acceptable salt as a chondroprotector; chondroprotector is sodium metabisulfite, while the pH of the medium is in the range from 3 to 2.

The technical result of the claimed invention is an increase in the stability of the drug, which consists in a significant reduction in the formation of degradation products of glucosamine and modification of ketoprofen, as well as an increase in the shelf life, at least up to 2 years.

Brief description of the drawings.

Fig. 1 - chromatograms of solutions of ketoprofen 2.5% with pH 3.0, showing the formation of ketoprofen impurities under stress conditions at a temperature of 60°C after 12 weeks of storage, with the following composition of solvents (from top to bottom): 1) 50% N-methylpyrrolidone (NMP ), 15% water, 35% PEG 400; 2) 50% NMP, 15% water, 35% propylene glycol (PG); 3) 50% NMP, 15% water, 35% diethylene glycol monoethyl ether (DME) (Transcutol® P); 4) 50% NMP, 15% water, 35% ethanol (96%); 5) 65% NMP, 5% water, 30% glycerin; 6) 85% NMP, 15% water.

Fig. 2 - UV absorption spectra of ketoprofen and ketoprofen impurities formed in solutions with PG, ethanol and glycerin, taken during chromatography of ketoprofen solutions.

Fig. 3 - chromatogram of a model solution of ketoprofen containing PG, obtained in the mass scan mode in the range of 120-500 m/z, where the peak with a retention time (Rt) of 6.945 min corresponds to ketoprofen; peak with Rt=9.064 min - ketoprofen PG ether, peak with Rt=9.749 min - ketoprofen PG ether.

Fig. 4 - mass spectrum of the peak with Rt=6.945 min (see Fig. 3) with M.m.=254 a.m.u.

Fig. 5 - mass spectrum of the peak with Rt=9.064 min (see Fig. 3) with M.m.=313 a.m.u.

Fig. 6 - mass spectrum of the peak with Rt=9.749 min (see Fig. 3) with M.m.=313 a.m.u.

Fig. 7 - plots of the solubility of ketoprofen in mixed solvents NMP-PEG 400 - water at temperatures of 6 and 25°C on the content of PEG 400 (at a constant NMP content of 18 wt.%).

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Fig. 8 - photographs of laboratory samples of the drug after storage for 1 year at a temperature of 25°C and different content of sodium metabisulphite: 0% (prototype drug), 0.25, 0.50, 0.75,

1.00%.

Fig. 9 - chromatogram of the test solution of the prototype drug, obtained under the conditions of quantitative determination of glucosamine impurities after storage of the prototype drug at a temperature of 25 ° C for 1 year, where the peak with Rt = 14.974 min corresponds to hydroxymethylfurfural (HMF), and peaks with retention times of 3.678 , 4.769, 5.096, 7.848, 8.258 and 10.731 min - unidentified degradation products of glucosamine.

Fig. 10 - chromatograms obtained under the conditions of quantitative determination of ketoprofen impurities, where 1) chromatogram of the reference solution, 2) chromatogram of the test solution of the drug containing PG and sodium metabisulphite, 3) chromatogram of the test solution of the drug containing PEG 400 and sodium metabisulphite, after 6 months of storage at 25°C.

Note. Peaks with Rt®5.134 min in chromatograms 1, 2 and 3 correspond to ketoprofen; peaks with Rt=6.515 min and Rt=7.020 min in chromatogram 2 correspond to ketoprofen PG esters. There are no peaks of foreign impurities on the chromatorogram of the test solution (1).

Fig. 11 - chromatograms obtained under the conditions of quantitative determination of glucosamine impurities, where: 1) chromatogram of a placebo solution; 2) chromatogram of the test solution of the drug after 6 months of storage at 25°C; 3) chromatogram of a standard solution containing HMF and methylpyrazine.

Note. Peaks with Rt=5.076 min and Rt=5.074 min in chromatograms 1 and 2 correspond to the drug base; peaks with Rt=1.093 min, Rt=1.227 and 1.930 min in chromatogram 2 correspond to unidentified impurities; the peak with Rt=6.171 min on the chromatogram of standard solution 3 corresponds to HMF, and the peak with Rt=7.869 min corresponds to methylpyrazine. There is no HMF peak on the chromatorogram of solution (2).

Fig. 12 - photographs of samples of the prototype drug (1) and the drug containing PEG 400 and sodium metabisulphite (2), after 1 year of storage at 25°C.

Study of the interaction of ketoprofen with hydrophilic solvents/

The preparation of Ketoprofen in the form of a gel is standardized in the monograph Ketoprofen Gel of the British Pharmacopoeia. So, in the gel of ketoprofen, which includes ethyl alcohol, with established pH limits from 5.0 to 7.5, it is proposed to normalize ethyl ester as foreign impurities of ketoprofen at a level of not more than 4%. That is, ketoprofen, being an organic acid, forms an ester with ethanol during storage. The composition according to the prototype contains propylene glycol, which also contains hydroxyl groups, and the pH of the gel is normalized in the range from 2.0 to 3.0. Therefore, the formation of ketoprofen modification products was studied depending on the composition of mixed solvents and the pH of solutions under stressful conditions - a temperature of (60±2)°C. A method for the quantitative determination of ketoprofen was previously developed and validated. The developed method met the requirements for the following validation indicators: specificity, correctness, linearity, precision [12]. A method for the quantitative determination of unidentified impurities of ketoprofen (for ketoprofen) was also developed and validated, which also corresponded to certain validation characteristics [12].

It was previously established that ketoprofen does not form modification products with N-methylpyrrolidone (NMP), which was used in all experimental samples of mixed solvents at a concentration of 50 wt.% to dissolve ketoprofen. Various solutions were prepared which contained 2.5% ketoprofen dissolved in mixed solvents containing 50% NMP, 15% water and about 35% either propylene glycol (PG) or glycerol or 96% ethanol or diethylene glycol monoethyl ether (DME). , Transcutol® P), or PEG 400. As a control sample, a solution containing 2.5% ketoprofen, 85% NMP and 15% water was prepared. Further, in each ternary system, the pH of the solution was adjusted to 3.0; 5.0 or 7.0 with either phosphoric acid or potassium hydroxide, after which these solutions were kept for 12 weeks at 60°C.

After the initial analysis of these solutions for the quantitative content of ketoprofen and the quantitative content of foreign impurities, it was found that no impurities of ketoprofen are formed during the preparation of experimental samples of solutions. Impurities were formed during the storage of solutions and when they were exposed to a high temperature of 60°C. Some results of analyzes of solutions of ketoprofen after their storage at a temperature of 60°C are presented in table. one.

As seen from the chromatograms in Fig. 1, an impurity of ketoprofen A (Rt®8.2 min) was formed in all solutions in concentrations from 0.78 to 0.92% (average 0.87%) (Table 1). The obtained results indicate that the formation of ketoprofen impurity A does not depend on the composition of solvents and pH of solutions, but is due to the impact on ketoprofen solutions of a stress factor - a high temperature of 60°C. When storing solutions at 25°C Ketoprofen impurity A was not formed.

In solutions containing PEG 400, PG, DME, ethanol or glycerol, i.e. solvents with hydroxyl groups, during storage at a temperature of 60 ° C, unidentified

- 3 039612 impurities. Thus, in the chromatograms of solutions containing PG or glycerol, two additional peaks are detected, and in solutions with ethanol and DME, one peak each (Fig. 1). In a solution with PEG 400, impurities also formed, but in a much smaller amount (Table 1). In the control solution containing NMP and water, unidentified impurities are not detected (Fig. 1, table. 1).

As examples in FIG. Figure 2 shows the UV spectra of ketoprofen and ketoprofen impurities obtained on a chromatograph with a diode array detector, formed in the presence of PG, ethanol or glycerol in solutions. These UV spectra are identical and have an optical absorption maximum at 255 nm, indicating the presence of the same chromophore group. It is obvious that the resulting impurities are derivatives of ketoprofen.

To identify these impurities, chromatograms of model solutions were obtained on a chromatograph with a mass detector, as well as mass spectra of ketoprofen and impurities with determination of their molecular weights (Table 1). An example of a chromatogram is shown in Fig. 3 and examples of mass spectra in FIG. 4-6.

As follows from FIG. 3-6, the mass spectrum of the peak with Rt=6.945 min corresponds to the mass spectrum of ketoprofen, and the peaks with Rt=9.064 min and Rt=9.749 min in the chromatogram shown in FIG. 3 belong to two esters of ketoprofen and PG, which have the same molecular weights of 312 amu. In solutions of ketoprofen containing glycerin, two glycerol esters of ketoprofen with the same molecular weights of 329 amu are also formed. The presence of two peaks of impurities in solutions of ketoprofen with PG and glycerol could be associated either with the formation of esters with different enantiomers of ketoprofen, since ketoprofen is a racemate, or with the interaction of ketoprofen molecules with different hydroxy groups of solvents. When using in solutions instead of racemate (ketoprofen) one enantiomer (dexketoprofen), two impurity peaks were also obtained on the chromatograms, which indicates the formation of esters of ketoprofen with different hydroxy groups of PG or glycerol.

In the presence of ethanol in a solution of ketoprofen, an impurity is formed with M.m. = 282 a.m.u., which corresponds to ketoprofen ethyl ether; in the presence of DME (Transcutol® R) - ester of DME and ketoprofen with M.m. = 370 a.m.u. (Table 1).

PEG 400 is a mixture of polyethylene glycols that have different molecular weights and contain terminal hydroxyl groups. When storing solutions of ketoprofen at a temperature of 60°C, respectively, PEG esters of ketoprofen with different molecular weights are formed, which appear as a single peak on the chromatogram (Fig. 1).

Table 1

The results of the analyzes of the solution at a pace] ) in ketoprofen after storage at 60°C Non-aqueous solvent pH M.m., a.u.m. nm Reducing the content of ketoprofen % Total content of esters, % Content of impurity A ketoprofen, % ethanol 3.0 282 255, 255 25.12 23.22 0.91 5.0 3.52 2.39 0.85 7.0 1.35 0.11 0.87 PG 3.0 312, 312 255, 255, 255 20.98 18.13 0.78 5.0 2.68 1.52 0.92 7.0 1.08 0.21 0.85 Glycerol 3.0 329, 329 255, 255, 255 22.69 20.42 0.86 5.0 8.24 6.71 0.89 7.0 3.56 3.01 0.87 DME (Transcutol® R) 3.0 370 255, 255 5.21 3.93 0.91 5.0 2.68 2.38 0.89 7.0 1.01 1.32 0.87 PEG 400 3.0 Do not define 255, 255 3.28 2.89 0.90 5.0 2.01 1.41 0.82 7.0 shared 1.02 Missing 0.80 ΝΜΠ 3.0 Missing 255 1.11 Missing 0.90 5.0 1.15 Missing 0.86 7.0 1.20 Missing 0.87

A drop in the concentration of ketoprofen in solutions during storage is accompanied by an increase in the content of ketoprofen esters (Table 1). By reducing the content of interaction products

-4039612 ketoprofen and hydrophilic solvents, the latter can be arranged in the following order: ethanol > glycerin >

PG > DME > PEG 400 > NMP. At the same time, NMP does not form esters with ketoprofen.

As follows from the data in Table. 1, with a decrease in the pH of the solutions, the content of esters of ketoprofen with ethanol, PG, glycerol, DME and PEG 400 increases. At pH 7.0, the decrease in the content of ketoprofen is insignificant, and the formation of esters is absent or minimal. At pH 5.0, the drop in the content of ketoprofen is greater on average by 2.4 times, and, accordingly, more ketoprofen esters are formed. In solutions having a pH of 3.0 and containing ethanol, PG and glycerin, the content of ketoprofen decreases by 21-25%, and the content of esters increases to 18-23%. At pH 3.0, the drop in the content of ketoprofen and the formation of esters are the smallest in the case of PEG 400. At the same time, the drop in the content of ketoprofen is less dependent on pH, and the content of the formed esters is approximately 6-10 times lower compared to solutions containing ethanol, glycerol and PG. It can be predicted that the formation of ketoprofen esters with PEG 400 at a temperature of 25°C will be insignificant for the stability of the drug.

According to the research results, for the development of a ketoprofen gel with an acidic environment, it is rational to use NMP, PEG 400 and water as solvents in optimal ratios for dissolving ketoprofen and glucosamine hydrochloride. That is, instead of the propylene glycol used in the prototype drug, PEG 400 should be used.

Study of the solubility of ketoprofen and glucosamine hydrochloride in the solvent system N-methylpyrrolidone - PEG 400 - water.

In table. 2 and in FIG. 7 shows the effect of the weight ratio between PEG 400 and water (at a constant content of NMP 18 wt.%) on the solubility of ketoprofen at temperatures of 6°C (refrigerator conditions) and 25°C (upper storage temperature limit).

table 2

Solubility of ketoprofen in mixtures of NMP (18 parts by weight) - PEG 400 - water, at temperatures of 6 and 25°C at different mass ratios between PEG 400 and water

Mass parts. The content of ketoprofen (% wt.) at temperatures: PEG 400 Water 6°C 25°C 0 60 0.11 0.31 ten fifty 0.30 1.02 20 40 1.41 3.77 24 36 3.48 8.73 28 32 5.96 17.66 32 28 8.86 26.36 36 24 13.94 33.29

As follows from the data presented in Table. 2 and in FIG. 7.2 g of ketoprofen are dissolved in a mixed solvent NMP - PEG 400 - water at temperatures of 6 and 25 ° C, if the content of PEG 400 is 22 and 15 g, respectively. the content of PEG 400. However, with an increase in the content of PEG 400, the solubility of the hydrophilic substance glucosamine hydrochloride decreases and the risk of its crystallization increases the more, the lower the temperature (Table 3).

Table 3

Crystallization of ketoprofen and glucosamine hydrochloride at different mass ratios between PEG 400 and water and at different temperatures

XtXt solutions: #1 #2 No. 3 #4 Xs 5 #6 No. 7 xs 8 Xs 9 xs 10 Xs 11 Glucosamine hydrochloride 5.0 Ketoprofen 2.0 ΝΜΠ 18.0 PEG 400 21.0 21.5 22.0 22.5 23.0 23.5 24.0 24.5 25.0 25.5 26.0 Purified water 39.0 38.5 38.0 37.5 37.0 36.5 36.0 35.5 35.0 34.5 34.0 Solution initially Solution at 25 °C, 1 day

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Solution at 25 °C, 2 days IM Solution at 25 °C, 3 days — Solution at 14 °C, 1 day one — Solution at 14 °C, 2 days one Solution at 14 °C, 3 days — Solution at 6 °C, 1 day W And Solution at 6 °C, 2 days W w w w — — — — eleven 111 w Solution at 6 °C, 3 days w AND! w AND! — Ugzh — P 111 111 PEG mass deviation from 23.75 g -11.6 -9.5 -7.4 -5.3 -3.2 -1.1 Thu +3.2 +5.3 +7.4 +9.5 Crystallization: Ketoprofen Glucosamine Notes: — no sediment; 1 single crystals; C precipitate; plentiful sediment

As follows from Table. 3, optimal for the solubility of both active ingredients in the dispersion medium of the gel is the content of PEG 400 from 23.5 to 24.0 g. At temperatures of 25 and 14 ° C, ketoprofen and glucosamine hydrochloride are soluble at a PEG 400 content of approximately 23.75 g ± 10% . With decreasing temperature, the solubility decreases; at a temperature of 14°C, single crystals of ketoprofen appear at a PEG 400 content of 21.0 g, and at a temperature of 6°C on the second day of storage of the solution, an abundant precipitate of ketoprofen is formed at a PEG 400 content of 22.5 g. At a temperature of 6°C, a precipitate of glucosamine hydrochloride formed when the content of PEG 400 is 25.0 g. With decreasing temperature, the area of mass ratios between PEG 400 and water, at which stable solutions of hydrophilic and hydrophobic active substances occur, narrows. At a temperature of 6°C, ketoprofen and glucosamine hydrochloride are soluble with a PEG 400 content of 23.75 g ± 3.2%. That is, there is a risk that when the mass ratio between PEG 400 and water shifts in one direction or another and at low temperature in a refrigerator, crystallization of one of the active substances will occur. Crystallization is reversible with increasing temperature; at a temperature of 15-25°C, the precipitated crystals dissolve. However, despite this, the temperature range from 15 to 25°C should be considered optimal for storing the drug, at which the physical stability of solutions of active substances is ensured.

Stability studies of the drug with a new composition of the solvent and the choice of a new composition of antioxidants.

In the reactor-homogenizer was developed experimental series of the drug with the composition according to the prototype. A series of the drug was packaged in tubes of 30 g and stored at a temperature of (25±2)°C.

However, after 6-12 months of storage, it became obvious that with this composition of glucosamine hydrochloride is unstable: the drug acquired a yellow color during storage (sample 1, Fig. 8), which was caused by a significant accumulation of degradation products of glucosamine hydrochloride, in particular hydroxymethylfurfural (HMF ). The chromatogram of the test solution of the prototype drug, showing the profile of the resulting decomposition products of glucosamine hydrochloride, is shown in Fig. 9. As can be seen from the chromatogram shown in FIG. 9, after 1 year of storage at a temperature of 25°C, HMF and six more unidentified impurities are formed in the preparation. The content of HMF was about 0.1%, which determined the yellow color of the gel.

For further studies, a series of the drug was made according to the prototype, in which PG was replaced by PEG 400. However, such a replacement did not bring cardinal changes regarding the stability of glucosamine hydrochloride and, accordingly, the change in the color of the drug during storage. In this regard, it was necessary to change the system of antioxidants, consisting of lidocaine hydrochloride, citric acid monohydrate and butylhydroxyanisole (as in the prototype), to a more effective antioxidant.

Sodium metabisulphite was chosen as a new antioxidant, which, on the one hand, provides an antioxidant effect, and, on the other hand, creates an acidic environment necessary for the stabilization of glucosamine hydrochloride (Table 4). The optimal content of sodium metabisulfite, at which the drug had a white color (sample 2, Fig. 8) and retained it during storage, and the gels had a pH of 2.35 to 3.02, is in the range from 0.2 to 0.3 % (Table 4). At higher levels of sodium metabisulphite, the pH of the gels was above 3.0 and the gels turned lemon yellow, indicating the formation of HMF impurities (samples 3, 4 and 5, Fig. 8).

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Table 4

Compositions (in percent) of some experimental samples of the drug with PEG 400 and sodium metabisulphite

Components Content (wt.%) in #1 compositions: #5 #2 No. 3 #4 Glucosamine hydrochloride 5.0 5.0 5.0 5.0 5.0 Ketoprofen 2.0 2.0 2.0 2.0 2.0 Sodium metabisulphite 0.200 0.225 0.250 0.275 0.300 Polyethylene glycol 20 1.0 1.0 1.0 1.0 1.0 cetostearyl ether Cetostearyl alcohol 5.0 5.0 5.0 5.0 5.0 poly(methyl vinyl ether maleic anhydride) decadiene 2.0 2.0 2.0 2.0 2.0 crosspolymer (trade name Stabileze® QM) Polyethylene glycol 400 23.7 23.7 23.7 23.7 23.7 X-methylpyrrolidone 18.0 18.0 18.0 18.0 18.0 Liquid paraffin (Vaseline oil) 5.0 5.0 5.0 5.0 5.0 Polydimethylsiloxane (trade hir. ten 1Ω 1Ω 1Ω 1Ω dimethicone 100) l, v ι,υ 1 ₉ and 1 ,ν Purified water up to 100.0 Gel pH: 2.35 \ 1 2.52 I 1 2.69 I 2.76 1 3.02

In a homogenizer reactor, an experimental batch of a preparation weighing 3 kg was produced, in which PG was replaced by PEG 400, and lidocaine hydrochloride, citric acid monohydrate, and butylhydroxyanisole were replaced by sodium metabisulfite (Table 5). The drug was packaged in tubes of 30 g and stored at (25±2)°C.

Table 5

The composition of the developed drug Component Compound in percentages g/3000 g Glucosamine hydrochloride (in terms of 100% dry matter) 5.0 150.0 Ketoprofen (in terms of 100% dry matter) 2.0 60.0 V-methylpyrrolidone 18.0 540.0 Polyethylene glycol 400 23.7 711.0 Polyethylene glycol 20 cetostearyl ether 1, oh 30.0 Cetostearyl alcohol 5.0 150.0 Stabileze® QM 2.0 60.0 vaseline oil 5.0 150.0 Dimethicone DM 100 1.0 30.0 Sodium metabisulphite 0.25 7.5 Purified water up to 100.0 up to 3000.0

At the same time, an experimental series of the drug was developed, which instead of PEG 400 contained PG. Samples of two batches of the drug after storage for 6 months at a temperature of 25°C were analyzed for the content of ketoprofen modification products. The chromatograms are shown in Fig. 10. As follows from the chromatograms shown in FIG. 10, in the preparation containing PG, already after 6 months of storage at a temperature of 25°C, two esters of ketoprofen and PG were formed, which correspond to peaks with Rt=6.515 min and Rt=7.020 min (Fig. 10); the total content of these PG esters of ketoprofen was 2.363%. On the chromatogram of the test solution of the preparation (1) (Fig. 10) containing PEG 400 and sodium metabisulfite, there were no peaks of foreign impurities.

As follows from the data presented in Fig. 9 and 11, the replacement of the antioxidants used in the drug prototype, 0.25% sodium metabisulphite leads to a change in the profile of impurities formed during storage of the drug at a temperature of 25°C. On the chromatogram of the test solution of the drug with sodium metabisulphite, stored for 6 months, there is no HMF peak, and the amount of unidentifiable impurities is reduced from six to three (Fig. 11). One of the impurities is a derivative of methylpyrazine, characterized by a relative retention time (relative to methylpyrazine) of about 0.25, and its accumulation characterizes the process of decomposition of glucosamine hydrochloride. If the prototype preparation quickly acquires a yellow color due to the formation of HMF during storage (sample 1,

- 7 039612 fig. 12), then the preparation containing 0.25% sodium metabisulphite retains a white color (sample 2, Fig. 12) or acquires only a barely noticeable yellowish tint during long-term storage due to the formation of small amounts of HMF. In the prototype preparation, the amount of HMF generated is approximately 10 times greater.

The results of analyzes of samples of a series of preparations containing sodium metabisulphite and PEG 400 are presented in table. 6.

Table 6

The results of drug analyzes during storage at (25±2)°C pH Foreign impurities (decomposition products), % Content in 1 g of the drug, mg Shelf life Conclusion Glucosamine hydrochloride (GG) Ketoprofen GG Ketoprofen Specification Norms: from HMF admixture single sum From from 2 years 2.0 <0.1% with RRt- i'm an impurity impurity 46.25 18.5 BEFORE 0.25 < <4.0% d<4.0 BEFORE BEFORE 3.0 1.0% % 53.75 21.5 The results of the analyzes of the drug: 2.48 but BUT BUT BUT 50.32 20.02 Initially Goodin 2.47 but 0.17 BUT BUT 49.54 20.18 3 months Goodin 2.48 but 0.32 but but 48.83 20.07 6 months Goodin 2.49 0.005 0.44 but but 48.24 19.93 9 months Goodin 2.47 0.009 0.50 0.14 0.14 48.00 19.85 12 Goodin months 2.46 0.015 0.52 0.24 0.24 47.88 19.75 eighteen Goodin 2.46 0.018 0.54 0.35 0.35 47.78 19.65 months Goodin 24 months Notes. one. HMF - hydroxymethyls >urfurol. 2. Impurity with RRt to 0.25 - product decomposition of glucosamine hydrochloride, which is a derivative of methylpyrazine; relative retention time of the peak of this impurity on the chromatogram (relative to methylpyrazine) is 0.25.

As follows from the analysis data presented in table. 6, the drug remains stable for two years in terms of critical quality indicators. Tendencies towards an increase in the content of foreign impurities and a decrease in the content of active substances are within the limits established in the specification. The prototype drug did not meet the specification requirements for the content of impurities and active substances (glucosamine hydrochloride and ketoprofen) after 1 year of storage.

Thus, the transdermal agent of the present invention has improved stability as well as a longer shelf life (at least two years).

A method for obtaining a transdermal composition according to the claimed invention.

Stage 1. Preparation of the base of the gel.

Vacuum homogenizer reactor No. 1 is sequentially loaded with pre-weighed

PEG 400 and Stabilese® QM. The loading of Stabilese® QM is carried out in portions (4-5 portions), between loads the mixture is mixed using a paddle mixer with scrapers with a rotation speed of about 30-60 rpm and a turbine mixer with a rotation speed of 1500-3000 rpm for 5-10 min. After loading the last portion of Stabilese® QM, the mixture is stirred with a paddle mixer at a speed of about 30-60 rpm and a turbine mixer at a speed of 1500-3000 rpm for 10-20 minutes until a homogeneous dispersion is obtained.

Cetostearyl alcohol and polyethylene glycol 20 cetostearyl ether pre-weighed on a balance are loaded into a vacuum homogenizer reactor No. 1 with a Stabilese® QM dispersion, heating is turned on and, while stirring with a paddle stirrer at a speed of 30-60 rpm, the mixture is heated to a temperature of (65± 5)°C. Upon reaching this temperature, the stirrers are turned on and the mixture is stirred using a paddle mixer with scrapers with a rotation speed of 30-60 rpm and a turbine mixer with a rotation speed of 1500-3000 rpm for 10-15 min until a homogeneous dispersion is obtained. Then turn on the cooling, and the mixture is stirred using a paddle mixer with scrapers at a speed of about 30-60 rpm to a temperature of (50±2)°C.

Stage 2. Preparation of a solution of ketoprofen.

Ketoprofen and N-methylpyrrolidone pre-weighed on a scale are loaded into container No. 2.

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If necessary, the mixture is heated to a temperature of 25-35°C and stirred with a stirrer until complete dissolution of ketoprofen.

Stage 3. Preparation of an aqueous solution.

Purified water, glucosamine hydrochloride and sodium metabisulfite are sequentially loaded into reactor No. 3, pre-weighed on a scale. If necessary, the mixture is heated to a temperature of 25-35°C and stirred with a stirrer until all components are completely dissolved. In the present method, glucosamine and/or ketoprofen and/or N-methylpyrrolidone and/or polyethylene glycol 400 and/or sodium metabisulphite are in the form of true solutions.

Stage 4. Preparation of the gel.

The solution of ketoprofen from stage 2 is loaded into the homogenizer reactor No. 1 with the paddle mixer with scrapers switched on. 05 MPa to -0.07 MPa and mix the mass using a paddle mixer with scrapers at a speed of 30-60 rpm for 10-20 minutes while cooling to a temperature of 35-37°C.

An aqueous solution from stage 3 is loaded into the homogenizer reactor No. 1 in portions in 4-5 steps with the paddle mixer with scrapers turned on at a rotation speed of 30-60 rpm. Upon completion of the loading of the aqueous solution, a vacuum is created with a depth of -0.05 to -0.07 MPa and the mass is mixed using a paddle mixer with scrapers with a rotation speed of 30-60 rpm and a turbine mixer with a rotation speed of 1500-3000 rpm in for 10-15 minutes until a homogeneous dispersion is obtained within 10-25 minutes. If necessary, the mass in the reactor-homogenizer No. 1 is cooled so that the temperature does not exceed 37-38°C.

Then turn on the cooling and the gel is cooled to a temperature of 20-25°C while stirring with a paddle mixer with scrapers with a speed of 30-60 rpm. Upon completion of mixing, if necessary, a vacuum is created in the reactor-homogenizer No. 1 with a depth of -0.05 to -0.06 MPa and the gel is kept without stirring for 1 hour for degassing.

After obtaining positive results of the analysis, the bulk gel is discharged from the homogenizer reactor No. 1 into an intermediate container for transportation, which is hermetically sealed.

Stage 5. Dosing of the gel into tubes.

In the intermediate container, the gel is transported to the packaging room and unloaded in parts into the bunker of the tube filling machine. The gel is dosed at 30 or 50 g in tubes. The air temperature in the production room during the packaging of the gel in tubes should be in the range from 18 to 25 ° C. Filled tubes are folded; at the same time, an imprint is applied to the tube indicating the series and expiration date.

Stage 6. Packaging of tubes in packs.

Each tube, together with instructions for medical use, is packed in a pack.

Stage 7. Packing packs into boxes.

Packs with tubes are packed in group packaging (boxes). The boxes are closed and a group label is glued to each of them.

Studies of pharmacological activity.

Further, comparative studies of the specific pharmacological action of the drug of the new composition were carried out. A comparison was made between a preparation containing 2% ketoprofen, PEG 400 and sodium metabisulphite (sample no. 1, see Table 5) and a marketed Fastum® gel containing 2.5% ketoprofen (sample no. 2).

Studies were carried out on 32 rats weighing 150-190 g (average 175 g). During the experiment, the animals were kept in a vivarium at 18–25°C, humidity 50–60%, natural day-night light, in standard plastic cages, on a standard diet [13].

Samples 1 and 2 were applied to the skin of the foot of rats up to the ankle joint in the amount of 100 mg, twice: 30 minutes before and immediately after the injection of the phlogogenic agent. The total amount of the drug applied to the rat was 200 mg, which, taking into account the average weight of rats (175 g), corresponded to 1.1 g/kg (calculated according to the dosage form). The choice of the dose was carried out experimentally by determining the number of samples of the compared preparations, which is necessary for their application in a thin layer on the entire surface of the ankle.

Exudative inflammatory edema was induced 30 min after the first application of the compared preparations by a subplant injection of 0.08 ml of a 0.5% aqueous solution of carrageenan into the right hind foot of rats [14].

The anti-inflammatory effect (AE) was assessed by the degree of inhibition of the increase in foot edema during the use of drugs in comparison with the control - a group of untreated animals. Foot volume was measured before (at baseline) and then 1, 3, and 5 hours after phlogistic injection using an electronic plethysmometer (mod. 7150, Hugo Basile, Italy). PE was calculated using the formula

PE \u003d [(ΔΥ _Κ - ΔΥ ₀ ) x 100%]: ΔΥ _Κ ,

- 9 039612 where Δν _κ and ΔV _ο - the average increase in the volume of the edematous foot in the control and experimental groups, respectively.

Pain sensitivity threshold (PST) was recorded 3 hours after phlogistic injection by the Randall-Selitto method with mechanical pain stimulation on an analgesimeter (Ugo Basile, Italy; mod. 7200). The analgesic effect (AE) was calculated from the increase in PBC in rats treated with the compared drugs, compared with untreated control animals, calculated by the formula

where F _ο and F _k - threshold pressure force on the foot (r) in the experimental and control groups, respectively.

Statistical processing of the results was carried out by methods generally accepted in pharmacology, calculating the average values of the indicators (X) and the standard error (S x). The significance of differences between the means was determined by Student's t-test. The probability of the results obtained was assessed at a significance level of at least 95% (p<0.05) [15, 16].

Research results.

The results of the study of anti-inflammatory and analgesic activity are presented in table. 7.

Table 7

Anti-inflammatory and analgesic effects of samples 1 and 2 in rats with carrageenan foot inflammation

Groups AV (ml) and PE (%) through ... h after the administration of carrageenan Total PE, %hch PCB (g) and AE (%) 3 hours after the administration of carrageenan one 3 5 PBCH, g AE, % AV, ml PE, % AV, ml PE, % △V ml PE, % Control, pathology untreated Sample No. 1 Sample #2 0.204±0.021 0.203±0.023 0.193±0.018 0.5 5.4 0.521±0.02 3 0.373* ±0.02 9 0.384* 28.4 26.3 0.551 ±0.021 0.346* ±0.026 0.360* ±0.017 37.2 34.7 94.8 95.4 151.9±28.8 204.4±56.6 198.8±47.8 34.6 30.9 ±0.02 0

Notes. 1. The preparations were applied to the skin twice (30 minutes before and immediately after the injection of phlogistic) at a total dose of 200 mg/animal.

2. ΔV, ml - increase in the volume of the inflamed foot of rats.

3. PE, % - anti-inflammatory effect.

4. PBC, d - threshold of pain sensitivity of the inflamed foot of rats.

5. AE, % - analgesic effect.

6. * - p<0.05 compared with control.

7. Number of animals in the group = 8.

Subplantar injection of a solution of carrageenan leads to the development of acute inflammatory exudative edema, which in untreated rats reaches a maximum by 3 hours. in the focus of inflammation with bradykinin and other kinins, by 3 h - with prostaglandins [17]. Thus, in the first 3 hours, the pathogenesis of carrageenan inflammation includes all the main mediators of pain and inflammation, which are targeted by the studied drugs.

Anti-inflammatory activity.

Double skin application of sample No. 1 (preparation with a ketoprofen content of 2.0% according to the present invention) and sample No. 2 (Fastum® gel with a ketoprofen content of 2.5%) has an anti-inflammatory effect, consisting in a decrease in the increase in the volume of the inflamed foot of rats compared with untreated control (Table 7). By 3 h, the increase in the volume of the inflamed foot in the groups of treated rats reached significant differences with the untreated control. The PE of the compared preparations was significantly manifested by 3 hours after the administration of carrageenan, amounting to 28.4% for sample No. 1, and 26.3% for sample No. 2. By 5 h, the severity of the anti-inflammatory effect of the drugs increased, amounting to 37.2 and 34.7%, respectively (Table 7). The total anti-inflammatory effects of both drugs were almost identical.

analgesic activity.

Double application of samples 1 and 2 also has an analgesic effect, consisting in an increase in the threshold of pain sensitivity (PBC) of the inflamed rat foot during its mechanical compression (Table 7). The analgesic effects of the drugs were comparable, however, the analgesic effect of sample No. 1 (34.6%) was slightly higher than the analgesic effect of sample No. 2 (30.9%).

Studies have shown that the composition of the present invention provides an anti-inflammatory effect and an analgesic effect comparable to those of Fastum® gel with a 20% reduction in the content of ketoprofen. In turn, a reduced concentration of ketoprofen will also reduce the risk of an ulcerogenic effect.

Bibliography.

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2. Kovalenko V. N., Bortkevich O. P. Osteoarthritis. Practical guide. Kyiv: Morion, 2005. 448 p.

3. Analgesics for Osteoarthritis: An Update of the 2006. Comparative Effectiveness Reviews / R. Chou, M. S. McDonagh, E. Nakamoto et al. Rockville (MD): Agency for Healthcare Research and Quality (US). 2011 Vol. 38. P. 1-148.

4. Pain and the safety problem of NSAIDs: monograph / A. V. Kuryata, T. K.

Lysunets, A. V. Zaichenko, A. V. Cherkasova. Dnepropetrovsk: Gerda, 2014. 84 p.

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Jul 2013 125(4 Suppl. 1). P. 7-18.

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2011. Jul 9; 71(10). P. 1259-1279.

9. Drug Delivery Across Physiological Barriers/ Ed. Silvia Muro. Pan Stanford Reference, 2016. 426 p.

10. Alkilani A.Z., McCrudden M.T.C., Donelly R.F. Transdermal Drug Delivery: Innovative Pharmaceutical Developments Based on Disruption of the Barrier Properties of the Stratum Comeum. pharmaceuticals. 2015. No. 7. p. 438470.

11. Topical and Transdermal Drug Delivery: Principles and Practice / Ed.

Benson H.A., Watkinson A.C. Wiley : Hoboken, NJ, USA, 2012. 464 p.

12. Guidelines for the validation of drug analysis methods / Ed. N.V. Yurgel, A.L. Mladentseva, A.V. Bourdein and others; Developers V.L. Bagirova, A.I. Grizodub, T.Kh. Chibilyaev and others - M.: Pharmaceutical industry, 2007. - 58 p.

13. Scientific and practical recommendations for the development of laboratory creations and robots with them / Yu.M. Kozhem'yakin, O.S. Khromov, M.A. Funenko, G.A. Sayfetdshova - K.: Avshena, 2002. - 156 p.

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15. Urbakh V.Yu. Statistical analysis in biological and medical research. -M.: Medicine, 1975. -295 p.

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CLAIM

Claims (4)

CLAIM 1. A transdermal agent for the treatment and prevention of diseases of the joints and soft tissues, containing a pharmaceutically required amount of a chondroprotector, a nonsteroidal anti-inflammatory agent, an antioxidant for a chondroprotector, a gelling agent capable of forming gels in an acidic environment, a mixture of nonionic emulsifiers of the 1st and 2nd kind and a mixture of solvents, where in it contains glucosamine or its pharmaceutically acceptable salt as a chondroprotector, ketoprofen as a non-steroidal anti-inflammatory drug, as a mixture of non-ionic emulsifiers - a mixture of cetostearyl alcohol and polyethylene glycol 20 cetostearyl ether, as a gel-forming agent capable of forming gels in an acidic environment - poly (methyl vinyl ether of maleic anhydride) decadiene crosspolymer, characterized in that the mixture of solvents contains N- - 11 039612 methylpyrrolidone, polyethylene glycol 400 and water, and the antioxidant for the chondroprotector is sodium metabisulfite, while the pH of the medium is in the range from 3 to 2. 2. A transdermal agent according to claim 1, characterized in that it additionally contains an emollient in the form of liquid paraffin and polydimethylsiloxane or in the form of a fatty oil or other lipophilic emollient. 3. Transdermal agent according to claim 2, characterized by the following composition, wt.%: glucosamine or its pharmaceutically acceptable salt - 2.0-7.0; ketoprofen - 0.5-4.0; N-methylpyrrolidone - 10.0-25.0; polyethylene glycol 400 - 21.0-26.5; polyethylene glycol 20 cetostearyl ether - 0.5-3.0; cetostearyl alcohol - 2.0-7.0; poly(methyl vinyl ether of maleic anhydride) decadiene crosspolymer - 0.5-3.0; liquid paraffin - 2.0-7.0; polydimethylsiloxane - 0.5-3.0; sodium metabisulfite - 0.2-0.3; water - up to 100.0. 4. Transdermal agent according to claims 2, 3, characterized by the following composition, wt.%: glucosamine hydrochloride (in terms of 100% dry matter) - 5.0; ketoprofen (in terms of 100% dry matter) - 2.0; sodium metabisulfite (in terms of 100% substance) - 0.25; N-methylpyrrolidone - 18.0; polyethylene glycol 400 (macrogol 400) - 23.7; polyethylene glycol 20 cetostearyl ether - 1.0; cetostearyl alcohol poly(methyl vinyl ether of maleic anhydride) - 5.0; decadiene crosspolymer - 2.0; liquid paraffin - 5.0; polydimethylsiloxane - 1.0; purified water - up to 100.0.