Rectal suppositories and their use

Abstract

The invention relates to rectal suppositories and their use for prolonged moistening of the large intestine walls.

Description

[0001] Rectal suppositories and their use

[0002] The present invention relates to rectal suppositories and their use for prolonged moistening of the large intestine walls. The invention is applicable in medicine.

[0003] Mucins (large, glycosylated proteins) secreted from intestinal epithelial cells form a gel, which then forms a mucus covering the intestinal wall from its lumen. Mucins are an important protection of the gastrointestinal lumen. Two layers of mucus are produced in the large intestine; the inner layer is attached, and the outer layer is less dense and unattached. In the colon, the outer layer of the mucus is the habitat of the microbiota, while the inner layer is impermeable to bacteria and is renewed every hour by surface goblet cells. However, the inner mucus layer can also become permeable to bacteria by several mechanisms including aberrations in the immune system. When bacteria arrive at the epithelial surface, the immune system is activated and inflammation is triggered, and this mechanism may occur in some types of ulcerative colitis (Johansson et al. 2013 - Johansson EV, Sjövall H, Hansson GC. The gastrointestinal mucus system in health and disease. Nat Rev Gastroenterol Hepatol. 2013 June; 10 (6):352-361. doi:10.1038 / nrgastro.2013.35). The effectiveness of most formulations administered locally depends largely on the time of their retention at the application site. In the case of formulations used on the mucous membranes, the phenomenon of mucoadhesion plays the key role, in which the wetting theory is crucial - it assumes there is a spontaneous propagation of the tested adhesive material on the surface of the mucous membrane and penetrating in its unevenness, which is the necessary condition for adhesion (Osmałek et al. 2017 - xOsmałek T, Froelich A, Jadach B, Snela A, Gadziński P, Ancukiewicz K. Znaczenie mukoadhezji w technologii farmaceutycznej. Część I. Opis zjawiska oraz techniki stosowane do jego oceny. FARMACJA WSPÓŁCZESNA 2017; 10:22-28). Rectal suppositories are known from Polpharma sold under the Debutir brand of the following composition: cocoa butter: 60%, sodium butyrate: 15%, sodium alginate: 12.5%, carboxymethylcellulose: 12.5%. Said product despite its advantages is still characterized by too low and short-lived wettability. According to the Polish Pharmacopoeia, the wettability of solids is usually determined by direct or indirect measurements of the contact angle. The contact angle between a liquid and a solid is the angle formed spontaneously when the liquid droplet is placed on the surface of the solid. For a given liquid, the wettable solids exhibit a low contact angle and the non-wettable - an angle of 90 degrees or more. Suppositories are sought with low wetting angle that is lasting longer than in the prior art. Unexpectedly, the technical problem described has been solved by the present invention.

[0004] The first subject of the invention is a rectal suppository containing sodium butyrate, acylglycerols, sodium alginate, and carboxymethylcellulose, characterized in that it contains from 40 to 50% by weight of acylglycerols, from 15 to 35% of sodium butyrate, and from 25 to 38% of sodium alginate and carboxymethylcellulose. Acylglycerols are a mixture of triacylglycerols, diacylglycerols and monoacylglycerols or a mixture of triacylglycerols and monoacylglycerols. The percentages given above, and the percentages given in the document herein, refer to the total weight of the suppository and are percentages by weight. In literature, the term "acylglycerols" is used interchangeably with the terms neutral fat or glycerides and refers to glycerol esters of fatty acids, see e.g. http: / / www.iupac.org / goldbook / G02647.pdf. Depending on the number of esterified hydroxyl groups of the glycerol, tri- (TAG), di- (DAG) and monoacylglycerols (MAG) can be distinguished. In the present invention, preferably the triacylglycerols are hydrogenated vegetable oil, preferably palm oil, diacylglycerols are diacylglycerol oil and the monoacylglycerol is glycerol monostearate and / or glycerol monolaurate. All of the above substances are commercially available. In a particular embodiment of the invention the acylglycerols are a mixture of triacylglycerols, diacylglycerols and monoacylglycerols. In another particular embodiment of the granulate of the invention, acylglycerols are a mixture of triacylglycerols and monoacylglycerols. Equally preferably, the suppository of the invention is characterized in that it comprises 50% by weight of acylglycerols, 25% by weight of sodium butyrate and 25% by weight of sodium alginate and carboxymethylcellulose, preferably 12.5% by weight of sodium alginate and 12.5% by weight of carboxymethylcellulose. More preferably, the suppository of the invention is characterized in that it comprises 40% by weight of acylglycerols, 25% by weight of sodium butyrate and 34% by weight of sodium alginate and carboxymethylcellulose, preferably 16.5% by weight of sodium alginate and 16.5% by weight of carboxymethylcellulose. Equally preferably, the suppository of the invention is characterized in that it comprises 50% by weight of acylglycerols, 15% by weight of sodium butyrate, 22.5% by weight of sodium alginate, and 12.5% by weight of carboxymethylcellulose. In a further preferred embodiment of the invention, the suppository is characterized in that it comprises 40% by weight of acylglycerols, 35% by weight of sodium butyrate and 25% by weight of sodium alginate and carboxymethylcellulose, preferably 12.5% by weight of sodium alginate and 12.5% by weight of carboxymethylcellulose.

[0005] The second subject of the invention is the use of the suppository as defined in the first subject of the invention for moistening of the large intestine walls.

[0006] The presented suppositories are characterized by a low contact angle lasting longer than in the prior art.

[0007] Exemplary embodiments of the invention are presented in the drawing in which Fig. 1 shows a chromatogram of the solution of suppository 1 in the medium (phosphate buffer at pH = 6.8), Fig. 2 - a chromatogram of the solution of suppository 2 (phosphate buffer at pH = 6.8), Fig. 3 - a chromatogram of the solution of tested suppository 3 in the medium (phosphate buffer at pH = 6.8) and Fig. 4 shows the contact angle © between the liquid and the solid as the angle formed spontaneously when the liquid droplet is placed on the surface of the solid.

[0008] Example 1. Method for preparing suppositories of the invention

[0009] 1.1 Method for preparing suppository 1 with the following composition: cocoa butter: 60%, sodium butyrate: 15%, sodium alginate: 12.5%, carboxymethylcellulose: 12.5%.

[0010] 1.1.1. Cocoa butter was introduced into the mixer

[0011] Temperature from 60°C to 65°C

[0012] Mixing speed: 45-65 rpm

[0013] Mixing time ≥ 1 hour

[0014] Temperature 60°C to 65°C

[0015] 1.1.2. Medium homogenization

[0016] Homogenization speed 3000 rpm - homogenization with bypass.

[0017] Mixing speed: 45-65 rpm

[0018] Temperature 60°C to 65°C

[0019] 1.1.3. Cooling of the mass cool the product to a temperature of 41°C

[0020] Mixing speed: 45-65 rpm

[0021] 1.1.4. Add sodium butyrate into the mixer

[0022] Mixing time ≥10 minutes

[0023] Mixing speed 45-60 rpm

[0024] Temperature from 39°C to 41°C

[0025] 1.1.5. Homogenization Homogenization speed 3000 rpm - homogenization with bypass. Homogenization time ≥ 5 minutes Mixing speed: 45-65 rpm

[0026] Temperature from 39°C to 41°C

[0027] 1.1.6. Add sodium alginate into the mixer

[0028] Mixing time ≥ 10 minutes

[0029] Mixing speed 45-60 rpm

[0030] Temperature from 39°C to 41°C

[0031] 1.1.7. Homogenization

[0032] Homogenization speed 3000 rpm - homogenization with bypass.

[0033] Homogenization time ≥ 10 minutes

[0034] Mixing speed: 45-65 rpm

[0035] Temperature from 39°C to 41°C

[0036] 1.1.8. Add carboxymethylcellulose into the mixer

[0037] Mixing time ≥ 10 minutes

[0038] Mixing speed: 45-60 rpm

[0039] Temperature from 39°C to 41°C

[0040] 1.1.9. Homogenization

[0041] Homogenization speed 3000 rpm - homogenization with bypass.

[0042] Homogenization time ≥ 10 minutes

[0043] Mixing speed: 45-65 rpm

[0044] Temperature from 39°C to 41°C

[0045] Portioning of the mass into suppository molds at temperatures from 39°C to 41°C

[0046] 1.2. Method for preparing suppository 2 with the following composition: cocoa butter: 50%, sodium butyrate: 15%, sodium alginate: 22.5%, carboxymethylcellulose: 12.5%. As in the example 1.1 for suppository 1, with the difference that the cooling of the mass in step 3 is carried out to a temperature of 55°C, the temperature in step 4 (introducing sodium butyrate to the mixer) is carried out at a temperature of 55 to 60°C, the homogenization in step 5 is carried out at a temperature from 55°C to 60°C, step 6 (introducing sodium alginate) at a temperature from 55°C to 60°C, step 7 (homogenization) at a temperature from 55°C to 60°C, step 8 (introducing carboxymethylcellulose) at a temperature from 55°C to 60°C, step 9 (homogenization) at a temperature from 55°C to 60°C, and portioning at a temperature from 55°C to 60°C.

[0047] 1.3. Method for preparing suppository 3 with the following composition: Cocoa butter: 40%, sodium butyrate: 25%, sodium alginate: 12.5%, carboxymethylcellulose: 22.5%.

[0048] As in the example 1.1 for suppository 1, with the difference that the cooling of the mass in step 3 is carried out to a temperature of 55°C, the temperature in step 4 (introducing sodium butyrate to the mixer) is carried out at a temperature of 55 to 60°C, the homogenization in step 5 is carried out at a temperature from 55°C to 60°C, step 6 (introducing sodium alginate) at a temperature from 55°C to 60°C, step 7 (homogenization) at a temperature from 55°C to 60°C, step 8 (introducing carboxymethylcellulose) at a temperature from 55°C to 60°C, step 9 (homogenization) at a temperature from 55°C to 60°C, and portioning at a temperature from 55°C to 60°C.

[0049] Example 2. Comparison of sodium butyrate release

[0050] Liquid chromatography with DAD detection was used as the analytical technique needed to assess the release of sodium butyrate. Prior to chromatographic analysis, the samples were subjected to a release process. The following indoor environmental conditions were maintained during the experiments:

[0051] - ambient temperature: 20°C±5°C

[0052] - relative ambient humidity: 65%±10%

[0053] 2.1 DESCRIPTION OF THE ANALYTICAL METHOD

[0054] Reference substances, chemical reagents and other materials The studies were performed using the reagents listed in Table 1.

[0055] Other materials:

[0056] - 10 mL, 20 mL, 2,000 mL volumetric flasks

[0057] - 1,000 mL volumetric flasks

[0058] - 10 mL, 100 mL, 2,000 mL beakers

[0059] - 1.5 mL vials with screw caps

[0060] - 2.0 mL eppendorf tubes

[0061] - automatic pipettes, adjustable volume 1000 μL, 10000 μL

[0062] - cellulose syringe filters

[0063] - Braun syringes 2 mL

[0064] - analytical balance with an accuracy of 0.001 g - Hanna Edge pH-meter

[0065] - ultrasonic bath

[0066] - flow release apparatus Equipment:

[0067] Liquid chromatograph, DAD detection: Nexera-i LC-2040C

[0068] Data processing and collection system: LabSolutions

[0069] Flow release apparatus Erweka USP 4 Flow-Through Cell DFZ II Chromatographic separation parameters

[0070] Table 2a. Chromatographic separation parameters

[0071] Table 2b. Gradient programme

[0072] Solutions for the analysis

[0073] Stock solution

[0074] 100 mg of sodium butyrate was weighed into a 2 mL volumetric flask. The flask was filled up to the mark with medium, mixed thoroughly. 6.7 μL of this solution was transferred to a 10 mL flask, the flask was filled to the mark with medium and mixed thoroughly. The solution was then filtered with a cellulose syringe filter of 0.22 μm pore diameter.

[0075] Preparation of the mobile phase

[0076] 4.800 g of sodium dihydrogen phosphate was weighed into a 2000 mL beaker, 1980 mL of water was added and stirred until dissolved. The pH=2.2 was then adjusted with phosphoric acid, the whole was transferred to a 2000 mL flask and filled up to the mark with demineralised water. Again, the pH was measured. The mobile phase was filtered.

[0077] Preparation of the medium - phosphate buffer (Na2HPO4) Approximately 13.1 g of sodium hydrogen phosphate and approximately 7.0 g of sodium dihydrogen phosphate were weighed, then 1900 mL of water was added and stirred until dissolved. The pH=6.8 was adjusted with phosphoric acid and filled up with demineralised water to 2000 mL. Again, the pH was measured.

[0078] Test solutions

[0079] One suppository of composition 1, 2 and 3, respectively, was placed in each of 6 test beakers filled with medium and thermostated (average suppository weight was 2000 mg) and analysed in the release apparatus. Description of the assay

[0080] Release

[0081] The release process was carried out using a medium. Solutions were collected according to the sampling schedule (Table 3) into eppendorf type tubes and then filtered into 1.5 ml vials using a cellulose syringe filter with 0.22 μm pore diameter.

[0082] Table 3. Programme parameters for the release process

[0083] Chromatographic analysis

[0084] Chromatographic analysis was performed by injecting the medium and the prepared solutions. Data were collected at λ = 210 nm.

[0085] Results

[0086] Table 4. Summary of results for sodium butyrate release from suppository 1 in medium.

[0087] Table 5. Summary of results for sodium butyrate release from suppositories 2 in medium.

[0088] Table 6. Summary of results for sodium butyrate release from suppositories 3 in medium.

[0089] The chromatograms are shown in the drawing in which Fig. 1 shows a chromatogram of suppository 1 solution in the medium (phosphate buffer pH=6.8), Fig. 2 - a chromatogram of tested suppository 2 solution (phosphate buffer pH=6.8) and Fig. 3 - a chromatogram of tested suppository 3 solution in the medium (phosphate buffer pH=6.8). The tests carried out indicate that the release of sodium butyrate from the test sample occurred to a negligible or minor extent in the case of suppository 2 and 3 compositions. The greatest amount of sodium butyrate after the full test duration was released in the case of suppository 1 composition - a suppository known from the prior art of the invention (sample 3.1.7). (approximately 5%).

[0090] Example 3 The purpose of the example was to determine whether and to what extent the composition of suppositories, in particular the content of sodium butyrate and of adjuvants in the form of sodium alginate and carboxymethylcellulose, affect their wettability, determined by the contact angle and the rate of change of this angle after the drop has settled on the surface (due to interactions with the surface of the suppository). According to the Polish Pharmacopoeia, the wettability of solids is usually determined by direct or indirect measurements of the contact angle. The contact angle between a liquid and a solid is the angle formed spontaneously when the liquid droplet is placed on the surface of the solid (Fig. 4). For a given liquid, wettable solids show a low wetting angle, while non-wettable solids show an angle of 90 degrees or more.

[0091] The test consisted of placing the suppositories of different compositions (3 suppositories of each composition) in a 37°C incubator until softened, the time being different for each composition and suppository. Once the correct consistency was obtained, each sample was thoroughly mixed with a spatula until the composition was as uniform as possible and placed between two microscope slides until' solidified. This was to obtain sample surface as smooth as possible to enable accurate measurements to be taken. The next step consisted of placing the sample, made as described above, in the measuring apparatus and placing a drop on its surface using a Hamilton micro-syringe, filled with 0.9% NaCl solution, installed in SEO's Phoenix Alpha apparatus. The measurement, carried out at room temperature, was repeated three times for each sample. Three samples were prepared for each of the compositions tested.

[0092] The measurement consisted of recording images of the droplets placed on the surface of the samples and determining the contact angle value using the Imaged > Plugins > Contact Angle function. In order to determine the change in the contact angle value over time, photographs of the same droplet were taken at intervals of 20 seconds, with the timing started from the moment the droplet was placed and ended at 60 seconds.

[0093] The suppositories were prepared according to Example 1.

[0094] Table 7 shows the composition of the prepared suppositories from 3.1.1 to 3.1.12.

[0095] Table 7. Suppository compositions

[0096] The aggregated results of the average values of contact angles and the rate of change of contact angles for the samples with different compositions are shown in Table 8. The values measured in times from 0 to 60 seconds from placing the droplet on the test surface are given.

[0097] Table 8

[0098] The highest contact angle values were obtained for sample compositions number 3.1.1 - 3.1.6, which did not contain sodium butyrate or contained small amounts (up to 5%) of this component, while for sample compositions number 3.1.7, 3.1.8, 3.1.10, 3.1.11, 3.1.12 and 3.1.13 containing from 15 to 35% sodium butyrate, significantly lower contact angle values were observed. This demonstrates a strong change in the character of the sample - from hydrophobic to hydrophilic - when shifting from low to high sodium butyrate content (and at the same time from high to low cocoa butter content). The minimum contact angle values were observed for samples with a sodium butyrate content of 25%. The rate of change of the contact angle after deposition of a saline droplet on the sample surface is an additional indicator of the surface's wettability and its interaction with the liquid. The data indicate that the rate of change of the contact angle increases with increasing sodium butyrate content. The tested parameter reaches the maximum value for a sodium butyrate content of 25%. The measurements of contact angles carried out for various times of contact of the liquid with the tested surface, as well as the calculations of the rate of change of contact angle values, clearly indicate that the sample composition, especially sodium butyrate content, significantly affects physicochemical properties of the sample, in particular their hydrophilicity and wettability. An increase in sodium butyrate content causes a decrease in the contact angle of sample surfaces and an increase in the rate of change of the contact angle. Both of these effects indicate that as the proportion of sodium butyrate increases, the surface of the suppositories becomes more and more hydrophilic and wettable. The effects described above are particularly visible in the sodium butyrate content range from 5 to 25% by weight. A further increase in the butyrate content (from 25 to 35%) does not result in a further increase in hydrophilicity observed, and even an unexpected decrease can be observed.

Claims

Claims1. Rectal suppository containing sodium butyrate, acylglycerols, sodium alginate and carboxymethylcellulose, characterised in that it contains 40 to 50% by weight of acylglycerols, 15 to 35% by weight of sodium butyrate and 25 to 38% by weight of sodium alginate and carboxymethylcellulose.

2. The suppository of claim 1, characterized in that it contains 50% by weight of acylglycerols, 25% by weight of sodium butyrate and 25% by weight of sodium alginate and carboxymethylcellulose, preferably 12.5% by weight of sodium alginate and 12.5% by weight of carboxymethylcellulose.

3. The suppository of claim 1, characterized in that it contains 40% by weight of acylglycerols, 25% by weight of sodium butyrate and 34% by weight of sodium alginate and carboxymethylcellulose, preferably 16.5% by weight of sodium alginate and 16.5% by weight of carboxymethylcellulose.

4. The suppository of claim 1, characterized in that it contains 50% by weight of acylglycerols, 15% by weight of sodium butyrate, 22.5% by weight of sodium alginate and 12.5% by weight of carboxymethylcellulose.

5. The suppository of claim 1, characterized in that it contains according 40% by weight of acylglycerols, 35% by weight of sodium butyrate and 25% by weight of sodium alginate and carboxymethylcellulose, preferably 12.5% by weight of sodium alginate and 12.5% by weight of carboxymethylcellulose.

6. Use of the suppository as defined in claims 1 to 4 for moistening the walls of the large intestine.

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