Granulate comprising a butyrate compound, its forms and uses

Abstract

Granulate comprising a butyrate compound, its forms and uses.

Description

The subject of the invention is a granulate comprising a butyrate compound, its forms and uses. The invention is applicable in medicine . From the state of the art, patent PL233287 is known, disclosing a granulate having in its composition a butyrate compound in an acylglycerol matrix, characterized in that it comprises a butyrate compound, preferably sodium butyrate, in an amount of 45 to 85 % by weight, acylglycerols in an amount of 15 to 55 % by weight, pharmaceutically acceptable excipients in an amount of 0 to 25 % by weight, wherein the acylglycerols constitute a mixture of triacylglycerols, diacylglycerols and monoacylglycerols, or a mixture of triacylglycerols and monoacylglycerols, wherein the percentage values refer to the total mass of the granulate. Small intestine bacterial overgrowth (SIBO) is defined as an increase in the amount of bacteria in the small intestine above normal values. The presence of SIBO is detected in 33.8 % of patients with gastroenterological complaints who underwent a breath test, and its presence is significantly associated with smoking tobacco, bloating, abdominal pain, and anemia. Also, proton pump inhibitor therapy is a significant risk factor for SIBO. The risk of SIBO increases with age and does not depend on gender or race. SIBO complicates the course of a number of diseases and may be of pathogenetic significance in the development of their symptoms. SIBO is significantly associated with functional dyspepsia, irritable bowel syndrome, functional abdominal bloating, functional constipation, functional diarrhea, short bowel syndrome, chronic intestinal pseudo-obstruction, lactase deficiency, diverticular disease and celiac disease, ulcerative colitis, Crohn's disease, cirrhosis, metabolic-associated fatty liver disease (MAFLD), primary biliary cholangitis, gastroparesis, pancreatitis, cystic fibrosis, gallstone disease, diabetes, hypothyroidism, hyperlipidemia, acromegaly, multiple sclerosis, autism, Parkinson's disease, systemic sclerosis, spondylarthropathy, fibromyalgia, asthma, heart failure (Efmerova et al. 2023) . Efmerova I, Maslennikov R, Poluektova E, Vasilieva E, Zharikov Y, Suslov A, Letyagina Y, Kozlov E, Levshina A, Ivashkin V. Epidemiology of small intestinal bacterial overgrowth World J Gastroenterol. 2023 Jun 14; 29(22) : 3400-3421. Published          online          2023          Jun 14. doi: 10.3748 / wjg.v29.122.3400 . Despite the high content of butyrate in the cited composition from the state of the art in patent PL233287, there is still a search for a form of administration that will transfer a larger amount thereof with such a release profile that almost entire amount thereof will be released in the ileum section or the large intestine section, depending on the location of the condition in the digestive tract. Surprisingly, the above-mentioned issue has been solved by the invention shown. The first subject of the invention is a granulate comprising a butyrate compound in an acylglycerol matrix, characterized in that it comprises - a butyrate compound, preferably either sodium butyrate or tributyrin, in the amount of 40 to 86 % by weight of the total mass of the granulate - acylglycerols in the amount of 14 to 58 % by weight of the total mass of the granulate, wherein they constitute a mixture of diacylglycerols and monoacylglycerols - pharmaceutically acceptable excipients in the amount of 0 to 10 % by weight of the total mass of the granulate. The term "acylglycerols" is used in the literature interchangeably with the terms "neutral fats" or "glycerides" and denotes esters of glycerol with fatty acids, see, for example, http: / / www.iu-pac.org / goldbook / G02647.pdf. Depending on the number of esterified hydroxyl groups of glycerol, a distinction can be made between tri- (TAG), di- (DAG) and monoacylglycerols (MAG). In the present invention, the diacylglycerols are preferably diacylglycerol oil, and the monoacylglycerol is preferably glycerol monostearate and / or glycerol monolaurate. All of the above substances are commercially available. A mixture of mono-and diacylglycerols with a suitable ratio which makes it useful in the context of the invention is also known under the name E-471. The second object of the invention is a unitary oral dosage form characterized by being filled with or manufactured based on a granulate, as defined in the first object of the invention. Preferably, the unitary oral dosage form according to the invention is selected from a hard capsule, a sachet, a stick pack, an ampule and a tablet, and more preferably it is a hard capsule . The third object of the invention is a granulate as defined in the first object of the invention with predominant release of the butyrate in the ileum for use in the treatment of small intestine bacterial overgrowth (SIBO). Unexpectedly, it turned out that using only a mixture of mono- and diacylglycerols without the addition of triacylglycerols makes it possible to obtain the expected release profile in the final segment of the human digestive system. Exemplary embodiments of the invention are shown in the drawing, in which Fig. 1 shows a standard curve for calculating the concentration of butyric acid in a sample, where the coefficient for calculating the butyric acid content a = 6.654 x ICT6 [mg / ml] and Fig. 2 shows a standard curve for calculating the concentration of sodium butyrate in a sample, where the coefficient for calculating the sodium butyrate content a = 6.183 x ICE6 [mg / ml] . Example 1 The method of encapsulation is any of the state of the art, for example the one disclosed in US4511584, wherein a composition of mono- and diacylglycerols is used as a carrier. The process involves providing a granular material based on a compound providing butyrate ions, mixing the granular material with a matrix containing mono- and diacylglycerol sources and preferably pharmaceutical acceptable excipients, heating the mixture to a temperature above the melting point of the acylglycerol component of the matrix, spraying the mixture in a cooling chamber with a temperature lower than the melting point of the acylglycerol component of the matrix, so that the latter solidifies around the granular material, forming a spatial coating system of said granular material. Further granulate forms comprise unitary dosage forms selected from a hard capsule, a sachet, a stick pack, an ampule and a tablet. The unitary forms according to the invention may be manufactured by conventional methods which are well known in pharmaceutical technology and which may be realized by using methods and means known from the state of the art, described, for example, in the textbook "Farmacja stosowana" ("Applied Pharmacy") edited by S. Janicki and A. Fiebig, PZWL, 4th edition. Example 2 In vitro digestion in a modified Bolsen's model according to the following variants: "  ---Variant STAGE A B c D E F EI pH 2, pepsin, 30 min X X llll X Eli      pH      6.8, pancreatin, 4 h lilll iiiiii liiii liiii lilll Illi EIII viscozyme, 6 h lilll ■III ■11 liiii ■II Illi All stages at 38°C. The stages reflect the conditions in the following segments of the digestive system: EI = stomach; Eli = duodenum and jejunum; EIII = ileum; Each assessment was performed in repetitions (1 sample = 3 assessments). Assessment is understood as carrying out the entire procedure, including determining the amount of released butyrate by HPLC technique. Description of HPLC conditions Separation parameters used to determine the amount of butyric acid: Mobile phases:                      Phase A:      2.5 mM methanesulfonic acid Phase B:                       100% acetonitrile Flow rate of the mobile phase:     0.450 ml / min Flow gradient:                0.0-0.6 min Phase A / Phase B (100 / 0) 0.6-7.2 min Phase A / Phase B (55 / 45) 7.2-9.6 min Phase A / Phase B (55 / 45) 9.6-10.6 min Phase A / Phase B (100 / 0) 10.6-15.0 min Phase A / Phase B (100 / 0) Column temperature:           30°C Injection volume:             3.2 pl Detection:                     210 nm Chromatograph:      Waters (pump 1525 with heater 1500; 2998 PDA; 2707 autosampler) The standard curves used are shown in figures 1 and 2, where in Fig. 1 the coefficient for calculating butyric acid content a = 6.654 x 10~6 [mg / ml] and in Fig. 2 the coefficient for calculating the content of sodium butyrate a = 6.183 x 10~6 [mg / ml]. Modified Bolsen's model Portions of 200 mg each of the test sample were placed in the appropriate number of Schott glass bottles (number of bottles = number of measuring points x 2 repetitions). After the completion of successive stages, 2 bottles each were separated for the assessment. STAGE I - reflecting the conditions of the gastric environment • 10 ml of 0.2 M HC1 was added to the preparation samples, and the pH was equalized to 2; • 1 ml of freshly prepared aqueous solution containing 25 mg of pepsin was added to the obtained mixture; • mixtures so prepared were incubated at 38°C for 30 minutes; • after the completion of incubation, 2 ml of the solution was collected from the bottles removed from the incubation in order to determine the amount of released butyric acid by the HPLC technique . STAGE IT - reflecting the conditions in the duodenum and jejunum • 10 ml of phosphate buffer (0.2 M, pH 6.8) + 5 ml of 0.6 M NaOH was added to the mixture after step I. pH was equalized to 6.8 with 1 M HC1 or 1 M NaOH; • 100 mg of pancreatin was added to each bottle and incubated at 38°C for 4 hours; • after the completion of incubation, 2 ml of the solution was collected from the bottles removed from the incubation in order to determine the amount of released butyric acid by the HPLC technique . STAGE ITT - reflecting the conditions in the ileum • 10 ml of 0.2 M EDTA, pH 6.8, was added to the mixture obtained in steps I and IT; • 1 ml of Viscozyme (Sigma) containing an enzymatic mixture of cellulase, arabinase, xylanase, pectinase and |3-glucanase was added; • incubating at 38°C for 6 hours; • after the completion of incubation, 2 ml of the solution was collected from the bottles removed from the incubation in order to determine the amount of released butyric acid by the HPLC technique . Description supplementing the methodology: Portions of 200 mg each of the preparations were weighed into Pyrex bottles, 10 ml of 0.2 M phosphate buffer with pH 6.8 and 3 ml of 0.6 M NaOH were added to each, the pH of the mixture was adjusted to 6.8 with 10 % HC1 solution and 100 mg of pancreatin was added to each. The bottles were capped, placed in an Elpin water bath shaker type 157 (shaking speed 200, amplitude 4) and incubated for 4 hours at 37 °C, simulating the digestion process in the jejunum and duodenum. After a predefined time, the pH of the mixtures was monitored, 1 bottle per tested preparation was removed from the bath, the reaction mixture was filtered (to separate the remaining microgranules) and the volume of the filtrate was measured. 1.5 ml of the mixture was collected from each sample, which was then centrifuged (10,000 rpm, 10 min) and filtered through PVDF hydrophilic filters (13 mm, 0.22 pm, by Biosens). The filtrate was assessed for butyric acid released from the samples during simulated intestinal digestion by HPLC method. To each of the remaining samples, 10 ml of 0.2 M EDTA solution with pH 6.8 and 1 ml of enzyme preparation Viscozyme® L were added. The bottles were capped, placed in a shaker type water bath, and incubated for 6 hours at 37 °C, simulating processes occurring in the ileum. SUBSTITUTE SHEET (RULE 26) The pH was monitored every 2 hours and adjusted, if necessary (a drop to 6.5), by the addition of NaOH. After 6 hours, the samples were removed from the bath, the reaction mixture was filtered and the volume of the filtrate was measured. 1.5 ml of the mixture was collected from each sample, which was then centrifuged (10, 000 rpm, 10 min) and filtered through PVDF hydrophilic filters (13 mm, 0.22 pm, by Biosens). The amount of butyric acid released from the samples during the total simulated digestion was determined by the HPLC method. Product composition: A - Sodium butyrate 75 %, mixture of MAG and DAG (E471) 25 % B - Sodium butyrate 75 %, mixture of MAG and DAG 15 %, calcium carbonate 5 %, sodium alginate 5 % C - Mixture of MAG and DAG (E471) 58 %, tributyrin 40 %, guar gum 2 % D - Mixture of MAG and DAG (E471) 48 %, tributyrin 42 %, candelilla 10 % E - Sodium butyrate 50 %, fully hydrogenated palm oil 47 %, glycerol monoalurate 1 %, sodium alginate 2 %, wherein it is a solution known from the state of the art, containing triacylglycerols in order to confront the presented invention F - Sodium butyrate 86 %, mixture of MAG and DAG (E471) 14 % The amount of sodium butyrate released during simulated digestion of preparations with sodium butyrate (mg / 200 mg preparation and %, respectively) - preparations A, B and E, respectively Sampling stage A B E F Stomach   (0.5 h) 2.85 ± 0.15 (1.9 %) 8.13 ± 0.34 (5.4 %) 5.10      ± 0.29 (5.1 %) 7.33      ± 0.36 (4.26 %) Stomach (0.5 h), duodenum and jejunum (4 h) 36.11     ± 0.25 (24.07 %) 71.76      ± 3.56 (47.8 %) 26.24    ± 1.13 (26.24 %) 41.50    ± 1.45 (24.13 %) Stomach   (0.5 h),   duodenum and jejunum (4 h) Ileum (6 h) 62.34     ± 0.93 (41.6 %) 79.90      ± 0.88 (53.3 %) 50.60 ± 0.15 (50.6 %) 160 ± 4.11 (93.02 %) Amount of butyric acid released during simulated gastrointestinal digestion of glycerol tributyrate preparations (mg / 200 mg preparation) - preparations C and D, respectively Sampling stage C D Stomach (0.5 h) 0.00 0.00 Stomach (0.5 h), duodenum and 23.48      ± 20.66    ± j ej unum (4 h) 1.13 0.24 (34.1 %) (28.6 %) Stomach (0.5 h), duodenum and 35.13      ± 31.36    ± j ej unum (4 h) 0.02 0.32 Ileum (6 h) (51.1 %) (43.4 %) In addition, the use of tributyrin as a source of butyrate ions, due to its preferable organoleptic parameters, facilitates the use of the preparation in a granulate form (sachets, stick packs) for direct oral administration or after adding to liquid food 5 (breast milk, yogurt, etc.) and use in pediatrics. In addition, glycerol tributyrate is not digested or dissociated in the acidic environment of the stomach, which further ensures that the granulate does not need to be protected, for example, by the use of a capsule. 10 Example 3 Comparison with a product of composition E, as in Example 2, manufactured based on the disclosure from patent PL233287 Effective dose of butyrate ions reaching the large intestine as a derivative of butyrate ion source, initial dose and release 15 profiles. A B C D E Amount of butyrate ions in mg as a source of butyric acid in the large intestine per 500 mg capsule 173.01 138.62 84.18 102.2 97.69 For the clinical effect in the treatment of, among others, inflammatory bowel disease, irritable bowel syndrome, diverticular disease, the amount of butyrate ions reaching the 20 large intestine is crucial. For preparations A and E, this difference is almost double. The dissociation constant pKa of butyric acid is 4.8, that is, for pH above 4.8 (typical for the small and large intestinal environment) it is dissociated. Example 4 5 Effective dose of butyrate ions reaching the ileum as a derivative of butyrate ion source, initial dose and release profiles . F Amount of butyrate ions in mg as a source of butyric acid in the ileum per a 500 mg capsule 234.03 It was found that the use of a higher amount of butyrate ion 10 source, i.e. above 80% in the granulate, leads to a preferable release profile, with predominant release in the ileum, which opens up new possibilities for the use of the product, including in SIBO, in which excessive bacterial growth in the terminal segment of the small intestine occurs. 15 In the case of SIBO, the amount of butyrate ions reaching the final segment of the small intestine, around the Bauhin's valve, is crucial.

Claims

1. A granulate comprising a butyrate compound in an acylglycerol matrix, characterized in that it comprises- a butyrate compound, preferably either sodium butyrate or tributyrin, in the amount of 40 to 86 % by weight of the total mass of the granulate- acylglycerols in the amount of 14 to 58 % by weight of the total mass of the granulate, wherein they constitute a mixture of diacylglycerols and monoacylglycerols- pharmaceutically acceptable excipients in the amount of 0 to 10 % by weight of the total mass of the granulate.

2. A unitary oral dosage form characterized in that it is filled with or manufactured based on a granulate, as defined in claim 1.

3. Granulate as defined in the first object of the invention for use in the treatment of small intestine bacterial overgrowth.

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