Fixed-combination pharmaceutical composition of a butyrate compound and probiotic bacteria for the treatment and prevention of symptomatic uncomplicated diverticular disease

Abstract

The invention relates to a fixed-combination pharmaceutical composition of a butyrate compound and probiotic bacteria for use in the treatment and prevention of symptomatic uncomplicated diverticular disease.

Description

[0001] Fixed-combination pharmaceutical composition of a butyrate compound and probiotic bacteria for the treatment and prevention of symptomatic uncomplicated diverticular disease

[0002] The invention relates to a fixed-combination pharmaceutical composition of a butyrate compound and probiotic bacteria for use in the treatment and prevention of symptomatic uncomplicated diverticular disease . The invention is useful in the field of medicine .

[0003] Symptomatic uncomplicated diverticular disease ( SUDD) is a standalone clinical entity characteri zed by recurrent abdominal symptoms attributable to diverticulosis in the absence of clinical , laboratory, or radiographic indicators of overt inflammation . No direct relationship between microfloral composition and diverticulosis has been established to date , with changes in the counts of speci fic bacteria and correlations between the disease symptoms and the signature of the microbiome being suggested by some preliminary data . Kvasnovsky et al . had examined fecal microbiota of 28 SUDD patients , comparing 15 patients with a history of diverticulitis with patients with no history of diverticulitis to identi fy certain correlations such as that of the higher intensity of bloating with the higher relative abundance of Rumi nococcus and lower representation of Roseburia species . Furthermore , fecal calprotectin levels were positively correlated with the relative abundance of Lactobacill us while pain scores were positively correlated with the relative abundance of Cyanobacteri um . (Kvasnovsky, 2018 ) . Di f ferences in the composition of fecal microflora in patients with acute diverticulitis at the time of diagnosis and patients with asymptomatic diverticulosis were also identi fied by Hullar et al . , who had observed discrepancies in the relative abundance of the species of genera Verrucomi crobia, Cori obacteria, Anaertruncus , Barnesi ella, Coprococcus , Subdoligranul um, Marvinbryantia, and Akkermansia (Hullar 2019) . Studies providing evidence for the potentially diagnostic signatures of intestinal and mucosal microflora were conducted in 14 patients without diverticulosis , 16 patients with asymptomatic diverticulosis and 8 SUDD patients (Barbara, 2017) . As suggested by the study results, fecal microflora of patients with diverticulosis and SUDD is deficient in Clostridium IV cluster. In addition, fecal microbiota of SUDD patients who had reported IBS-like symptoms was found to be depleted with regard to the representatives of Lactobacilaceae and Clostridium cluster IX species. The latter bacteria are known to produce propionate which is known to have a number of health-promoting effects, including anti-lipogenic, cholesterol-lowering, anti-inflammatory and anti-cancer effects. Therefore, depletion in these bacterial counts can potentially be linked to the negative effects of SUDD (Flint, 2012) . Representatives of the mucin-decomposing Akkermansia genus present with anti-inflammatory and anti-obesity properties (Karlsson, 2012) , and their depletion had been associated with intestinal inflammation (Fisher CK 2014) ; in SUDD patients, Akkermansia species were less abundant within the diverticular region as compared to the more distant locations (Barbara, 2017) . The proposed management of SUUD involves controlling the symptoms and reducing the risk of complications by means of fiber-rich diet, antibiotics acting locally within the gastrointestinal tract (e.g. rifaximin) and mesalazine, as well as probiotics (low-grade recommendation, lack of sufficient clinical evidence) ; elective surgery may be considered in patients with treatment failure and persistently poor quality of life (Comparato et al., 2007, Spiegel et al. 2015, Tursi et al. 2020, Justin et al. 2020, Mari et al. 2021) . The 2019 international consensus on diverticulosis and diverticulosis was indicative of some evidence being available for the potential effective reduction of certain symptoms in SUDD patients by the use of probiotics [EL: 3a; RG: B] (Tursi et al. 2019) . However, a systematic review could not demonstrate the efficacy of probiotics in diverticular disease or SUDD due to the high heterogeneity and poor quality of the data obtained (Lahner et al. 2016) ; no recommendations to use probiotics in SUDD have been provided in other guidelines (Hall et al. 2020, Cuomo et al. 2014) . Therefore, broader, randomized trials are still needed before probiotics can be recommended for the treatment of SUDD based on relevant evidence (Binda et al. 2015) . This position has been maintained in a 2023 review paper, the authors concluding that due to their capability of modifying microfloral composition and their anti-inflammatory effects, probiotics could potentially effectively reduce symptoms in SUDD patients, albeit adequate evidence of their usefulness is lacking (Calini et al. 2023) . Lahner et al. (2012) evaluated the effect of a fiber-rich diet alone and in combination with Lactobacillus paracasei B21060 supplementation on symptoms of uncomplicated diverticulitis (other than SUDD) to show that a reduction in short-term abdominal pain and abdominal distension was observed that in both groups of patients while long-term abdominal pain relief was observed only in the group of patients in the probiotics group (Lahner. 2012) . The results of clinical trial NCT02115867 as published by Kvasnovsky et al. (2017) suggest that the use of a liquid probiotic cocktail (Lacti caseibaci llus rhamnosus NCIMB 30174, Enterococcus faecium NCIMB 30176, Lactobacillus acidophilus NCIMB 30175, Lactiplantibacillus plantarum NCIMB 30173) did not significantly reduce abdominal pain while significantly reducing the incidence of the remaining four symptoms associated with chronic non-acute symptomatic diverticular disease (Kvasnovsky, 2017) . The results of clinical trial NCT01609751 involving administration of Lactobacillus easel Shirota strain to patients with symptomatic uncomplicated diverticular disease as published by Nichols et al. (2020) proved ambiguous (Nichols. 2020) . As demonstrated in three clinical trials, patients with SUDD present with better remission maintenance when treated with Lactobacillus species, especially in combination with mesalazine (Tursi et al. 2006, 2013, 2008) . Further studies evaluated the efficacy of Lactobacillus acidophilus, L. helveticus and Bi fidobacteri um spp . 420 or Lactobacillus paracasei in patients with uncomplicated DD to observe reduction in abdominal pain and bloating (Annibale 2011, Lamiki 2010) . Another study (Fric & Zavoral 2003) compared the currently unpracticed treatment of diverticular disease symptoms - dichlorquinoline (an antibacterial agent acting locally within the intestine) and activated charcoal, with the use of Escherichia coll Nissle strain, the results showing a statistically longer periods of remission of abdominal pain, irregular bowel movements, and bloating when the bacterial strain was used. A review by Ojetti et al. (2018) evaluated a total of 13 clinical trials, including 3 double-blind, randomized, placebo-controlled trials, 6 randomized open-label trials, and 4 non-randomized open-label trials. In the conclusions, the reviewers stated that although most of the studies were still preliminary, the available data indicate the possibility of clinical use of certain probiotic strains in the treatment of diverticular disease at any stage (Ojetti, 2018) . A more recent review by Picconi et al. (2021) still lacks sufficient data for solid conclusions being drawn with regard to the efficacy of probiotics in the treatment of diverticular disease symptoms. In the only study published so far to discuss the use of butyrate ions in diverticular disease, the authors (Krokowicz et al. 2014) had evaluated the effect of sodium butyrate on the risk of diverticulitis episodes in patients with asymptomatic diverticular disease, i.e. in a pathological entity different than SUDD. The results of the study were indicative of a statistically significant reduction in the risk of diverticulitis in patients receiving sodium butyrate as compared to the placebo group; sodium butyrate has also had a beneficial impact on the subjective assessment of the quality of life of patients . Therefore , an oral formulation of a pharmaceutical composition ef fective in the treatment of symptomatic uncomplicated diverticular disease ( SUDD) is still being sought . Unexpectedly, this problem has been solved by the present invention .

[0004] The invention relates to a fixed-combination pharmaceutical composition of a butyrate compound and probiotic bacteria for use in the treatment and prevention of symptomatic uncomplicated diverticular disease . For the purposes of this invention, the terms "treatment" and "prevention" should be understood as any action that can contribute to the improvement of the patient ' s condition, including administration of a drug, supplement , or food for special medical purpose . The term " fixed combination" is well-established within the field and relates to two or more active ingredients contained in a single pharmaceutical formulation . Another preferred embodiment of the invention is characteri zed in that the butyrate compound is sodium butyrate or tributyrin . An equally preferred fixed-combination pharmaceutical composition is characteri zed in that the daily dose of butyrate ions is 246 to 486 mg . The term "butyrate ion" is understood as the molar mass of the butyrate moiety against the molar mass of the entire compound used as the source of butyrate ions following its release in the aquatic environment ( in the case of sodium butyrate and tributyrin, the molar mass of butyrate ions accounts for about 79% and 86% of the total molar mass , respectively) . In another preferred embodiment of the invention, the composition of the invention is characteri zed in that the daily dose of probiotic bacteria is 10 . 4 x 109CFU to 16 . 4 x 109CFU, preferably administered in two doses of 5 . 2 x 109to 8 . 2 x 109CFU per dose . Dosing regimens di f ferent from the twice-daily regimen and summing up to the total daily dose , such as regimens involving administration of three or four divided doses , are also envisioned . An equally preferred fixed- combination pharmaceutical composition for use according to the invention is characterized in that the probiotic bacteria are selected from the group comprising Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus reuterii, Bifidobacterium infantis. In the context of a group of probiotic bacteria, the tern "contains" can also be understood as "consists of". More preferably, the fixed-combination pharmaceutical composition for use according to the invention is characterized in that it additionally contains probiotic yeast, preferably Saccharomyces cerevisiae var. boulardii in the quantity of 4.0 x 109CFU per daily dose. As in the case of bacteria, different dosage regimens not exceeding the indicated daily dose are envisioned. More preferably, the fixed-combination pharmaceutical composition contains acylglycerols selected from the group comprising mono-, di- and / or triacylglycerols of plant origin. Most preferably, composition according to the invention is characterized in that the butyrate compound is in the form of acylglycerol-coated microgranules .

[0005] Unexpectedly, combination of butane ions with a probiotic cocktail, in particular with L. rhamnosus and L. plantarum strains, was shown to bring about statistically significant benefits with regard to the symptoms of diverticular disease, the Wexner scale scores, and the quality of life parameters (statistical significance to be confirmed) in patients, as well as with regard to effective colonization with probiotic strains.

[0006] Example embodiments of the invention are presented in the figures, with figure 1 presenting the results of the statistical analysis of the study data showing significant differences in the quality of life and general health categories between visits VI and V3; figure 2 presenting the results of the statistical analysis of the study data showing significant differences in diverticular disease symptoms between visits VI, V3 and V4; figure 3 presenting the results of the statistical analysis of the study data showing significant differences in Wexner stool incontinence scores between visits VI and V4; figure 4 presenting the results of the statistical analysis of the study data showing significant differences in colonization with Product 1- administered strains between visits VI, V3 and V4; figure 5 presenting the results of the statistical analysis of the study data showing significant differences in diverticular disease symptoms between visits VI, V2 and V3; figure 6 presenting the results of the statistical analysis of the study data showing significant differences in colonization with Product 2- administered strains between visits VI, V2 and V3; and figures 7 and 8 presenting the results of the study presented in Example 3.

[0007] Example 1

[0008] The product consists of an encapsulated source of butyrate ions, i.e. sodium butyrate, and 2 to 4 live probiotic strains (preferably Lactobacillus rhamnosus (ATCC 53103) GG, Lactobacillus plantarum LP01, Lactobacillus reuterll LRE02, Bl fldobacterl um Inf antis B102) formulated together as capsules, wherein the sodium butyrate is encapsulated using any of the methods known in the art, for example as disclosed in US 4,511,584, and the probiotic strains are encapsulated in a lipid matrix of plant origin or, preferably, wherein sodium butyrate and the probiotic strains are encapsulated together using an acylglycerol carrier of plant origin. Upon formulation, each capsule contains at least 5.2 x 109live bacterial cells, the quantitative composition being given in Table 1. The recommended dosage is two capsules administered as a single dose or as two divided doses daily. A decrease in the number of live colonies of bacteria during the storage of capsules is a known phenomenon . Table 1 : Quantitative and qual itative composition of Product 1

[0009] Alternatively, for the implementation of the aforementioned variant , the fixed-combination pharmaceutical composition according to the invention may have the following composition per dosage unit for twice daily administration :

[0010] The study was carried out in 23 patients [18 (78.3%) women and 5 (21.7%) men] . The average age of the patients was 69.317.2 years, the median age was 68.5 years, and the range spanned from 50.4 to 84.9 years A total of 19 patients (82.6% of the group) were aged 40-75 years 1.1. Clinical Study Design A prospective , single-center study .

[0011] All the patients enrolled in the study had symptomatic uncomplicated diverticular disease as confirmed by endoscopy, with a history of an inflammatory episode within 6 months prior to the study .

[0012] The duration of the study was 12-14 weeks for each participant attending a total of 5 visits (V0-V4 ) . Study flow diagram :

[0013] Product administration period

[0014] VO - veri fication of inclusion / exclusion criteria ; signing of informed consent form; instructions for patients regarding sel fcollection of feces .

[0015] VI - first study visit ( 0-14 days after veri fication of inclusion criteria ) : beginning of Product 1 administration, general medical examination; assessment of clinical signs such as bloating and fecal incontinence using the Wexner scale ; severity of diverticular disease symptoms ; fecal sample for coloni zation study; quality of li fe assessment using an obj ective SF-36 questionnaire

[0016] V2 - second study visit ( 6 weeks after inclusion) : general medical examination; control of adverse reactions ; subsequent portions of the study Product 1 being given out . V3 - third study visit ( 9 weeks after inclusion) : completion of Product 1 administration, general medical examination; control of adverse reactions ; fecal sample for coloni zation study; intensity of diverticular disease symptoms ; quality of li fe assessment using the standardi zed SF-36 questionnaire ;

[0017] V4 - fourth study visit ( 12 weeks after inclusion and 3 weeks after last administration of the probiotic composition) : general medical examination; presence of clinical signs such as gasses and fecal incontinence using Wexner scale ; severity of diverticular disease symptoms ; fecal sample for coloni zation study; quality of li fe assessment using an obj ective SF-36 questionnaire

[0018] 1 . 2 . The method for determination of patients ' intestinal coloni zation with strains administered with Product 1 having the composition as disclosed in Table 1 .

[0019] Fecal samples sel f-collected by patients into test tubes filled with a buf fer to protect the genetic material using sampling spoons attached to the tube caps prior to visit VI , V3 , and V4 ( in duplicates ) . Metagenomic DNA isolated by column-based extraction, concentration measured by spectrophotometry, stored at -20 ° C until submitted to qPCR assay . The presence of each strain within the preparation veri fied by 2 reactions : 2 pairs of primers to detect species-speci fic and strain ( or a group of strains ) -speci f ic markers , the identity of known genomic fragments being >98 % (based on the BLAST analysis of the NCBI GenBank database content ) ; the result considered positive only in the case of both products being obtained (veri fication of softening temperature by comparison with a positive control reaction involving the genomic DNA of the desired strain as performed in parallel to the test reaction) . In addition, a positive control with universal 16S rRNA starters and a negative control without matrix were run for each assay

[0020] 1.3 Results of the study

[0021] The presented results are indicative, in a consistent and statistically unambiguous manner, of a synergistic effect in relation to the tested parameters, said synergistic effect being impossible to obtain, on the basis of literature data, from independent administration of a source of butyric ions and probiotic bacteria. The results are characterized by high statistical significance in relation to the results of the patients' subjective assessments, the clinical parameters, and the ability of bacteria to permanently colonize the large intestine. The technical problem solved by the invention leads to a clear and unexpected improvement in the condition of patients with SUDD

[0022] The results are shown in Figures 1 to 4.

[0023] 1.4 Statistical analysis

[0024] The statistical analysis was carried out using the StatisticalS .3 package (StatSoft Polska, Dell Statistica Partner) . Quantitative data were summarized using descriptive statistics (mean ± SD, median and range) . The distribution of each quantitative variable was verified for compliance with normal distribution using the Shapiro-Wilk's test. Qualitative data have been summarized as percentage ratios. To check the efficacy of the probiotic composition in the supportive treatment, product assessments were examined for statistically significant differences between visits: VI (before the start of probiotic treatment) , V3 (9 weeks after the start of the treatment) , and V4 (3 weeks after the completion of the treatment) . The x2test or the Fisher's exact test was used for qualitative data while the Friedman' s non-parametric ANOVA test was used for quantitative data. The level of statistical significance was established at a=0.05. If statistically significant differences were found between consecutive visits, an additional examination was made to identify potential differences between visits VI and V3 and between visits VI and V4. The Bonferroni correction was applied due to the multiple comparisons; in this case, the statistical significance level was established at 0.025. In addition, a non-parametric Mann- Whitney test was performed to assess the differences between the study subgroups in terms of the clinical symptoms of diverticular disease. Correlation analysis was carried out using the Spearman's or the Pearson's correlation coefficient.

[0025] Example 2

[0026] The product consists of an encapsulated source of butyrate ions, i.e. tributyrin, and 2 to 4 live probiotic strains (preferably Lactobacillus rhamnosus (ATCC 53103) GG, Lactobacillus plantarum LP01, Lactobacillus reuterll LRE02, Bl fldobacterl um Inf antis B102) formulated together as capsules, the encapsulation method being chosen from those known in the art, e.g. as revealed in US4511584. Each capsule contains a minimum of 8.2 x 109CFU of live bacterial cells, the quantitative composition being given in Table 2.

[0027] The recommended dosage is two capsules administered as a single dose or as two divided doses daily.

[0028] The use of simultaneous encapsulation of the source of butylene ions and strains of probiotic bacteria results in beneficial protection of bacteria against the enzymatic environment of the small intestine and the release of bacteria in the light of the large intestine, promoting their proper colonization, additionally supported by the presence of butylene ions. Independently, microcoating increases the stability of the composition and increases the bacterial survival time during storage in the refrigerator (4-6°C) . The bacterial counts amounted to 8.2 x 109CFU at baseline , 8.0 x 109CFU at 4 months, and 9.4 x 108CFU at 10 months. Table 2: Quantitative and qualitative composition of Product 2.

[0029] The study was carried out in 15 patients [10 (66.6%) women and 5 (33.3%) men] . The average age of the patients in the study group was 65.2 ± 4.4 years The duration of the study was 9 weeks, including a 6-week product administration period and a 3-week wash-out period .

[0030] 2 . 1 . Study Design

[0031] The duration of the study was 9---11 weeks for each participant attending a total of 4 visits (V0-V3 ) . Study flow diagram: 0-14 days 6 weeks 3 weeks

[0032] Product administration period

[0033] VO - veri fication of inclusion / exclusion criteria ; signing of informed consent form; instructions for patients regarding sel fcollection of feces .

[0034] VI - first study visit ( 0-14 days after veri fication of inclusion criteria ) : beginning of Product 2 administration; general medical examination; severity of diverticular disease symptoms ; fecal sample for coloni zation study;

[0035] V2 - second study visit ( 6 weeks after inclusion) : completion of Product 2 administration, general medical examination; control of adverse reactions ; fecal sample for coloni zation study; intensity of diverticular disease symptoms ;

[0036] V3 - third study visit ( 9 weeks after inclusion and 3 weeks after last administration of the probiotic composition) : general medical examination; control of adverse reactions ; intensity of diverticular disease symptoms ; fecal sample for coloni zation study . 2.2. The method for determination of patients' intestinal colonization with strains administered with Product 2 having the composition as disclosed in Table 2.

[0037] Fecal samples self-collected by patients into test tubes filled with a buffer to protect the genetic material using sampling spoons attached to the tube caps prior to visit VI, V2, and V3 (in duplicates) . Metagenomic DNA isolated by column-based extraction, concentration measured by spectrophotometry, stored at -20°C until submitted to qPCR assay. The presence of each strain within the preparation verified by 2 reactions: 2 pairs of primers to detect species-specific and strain (or a group of strains ) -specif ic markers, the identity of known genomic fragments being >98% (based on the BLAST analysis of the NCBI GenBank database content) ; the result considered positive only in the case of both products being obtained (verification of softening temperature by comparison with a positive control reaction involving the genomic DNA of the desired strain as performed in parallel to the test reaction) . In addition, a positive control with universal 16S rRNA starters and a negative control without matrix were run for each assay

[0038] 2.3. Results. The presented results are indicative, in a consistent and statistically unambiguous manner, of a synergistic effect in relation to the tested parameters, said synergistic effect being impossible to obtain, on the basis of literature data, from independent administration of a source of butyric ions and probiotic bacteria. The results are characterized by high statistical significance in relation to the results of the patients' subjective assessments, the clinical parameters, and the ability of bacteria to permanently colonize the large intestine. The technical problem solved by the invention leads to a clear and unexpected improvement in the condition of patients with SUDD.

[0039] The results are shown in Figures 5 and 6.

[0040] 2.4. Statistical analysis

[0041] The statistical analysis was carried out using the Statistical! .3 package (StatSoft Polska, Dell Statistica Partner) . Quantitative data were summarized using descriptive statistics (mean ± SD, median and range) . The distribution of each quantitative variable was verified for compliance with normal distribution using the Shapiro-Wilk's test. Qualitative data have been summarized as percentage ratios. To check the efficacy of the probiotic composition in the supportive treatment, product assessments were examined for statistically significant differences between visits: VI (before the start of probiotic treatment) , V3 (9 weeks after the start of the treatment) , and V4 (3 weeks after the completion of the treatment) . The x2test or the Fisher's exact test was used for qualitative data while the Friedman' s non-parametric ANOVA test was used for quantitative data. The level of statistical significance was established at a=0.05. If statistically significant differences were found between consecutive visits, an additional examination was made to identify potential differences between visits VI and V3 and between visits VI and V4. The Bonferroni correction was applied due to the multiple comparisons; in this case, the statistical significance level was established at 0.025. In addition, a non-parametric Mann- Whitney test was performed to assess the differences between the study subgroups in terms of the clinical symptoms of diverticular disease. Correlation analysis was carried out using the Spearman's or the Pearson's correlation coefficient.

[0042] Example 3. The product consists of an encapsulated source of butyrate ions, i.e. tributyrin, and 2 to 3 live probiotic strains (preferably Lactobacillus rhamnosus (ATCC 53103) GG, Lactobacillus plantarum LP01, Saccharomyces cerevisiae var. boulardii) (CNCM 1-3799) formulated together as capsules, the encapsulation method being chosen from those known in the art, e.g. as revealed in US4511584. Each capsule contains a minimum of 1.0 x IO10CFU of live bacterial cells, the quantitative composition being given in Table 3.

[0043] The recommended dosage is two capsules administered as a single dose or as two divided doses daily.

[0044] The study was carried out in 15 patients [8 (53.3%) women and 7 (46.7%) men] . The average age of the patients in the study group was 63.2 ± 3.9 years The duration of the study was 9 weeks, including a 6-week product administration period and a 3-week wash-out period. 0-14 days 6 weeks 3 weeks

[0045] Product administration period

[0046] 3.1. Study Design

[0047] The duration of the study was 9-11 weeks for each participant attending a total of 4 visits (V0-V3) . Study flow diagram:

[0048] VO - verification of inclusion / exclusion criteria; signing of informed consent form; instructions for patients regarding selfcollection of feces.

[0049] VI - first study visit (0-14 days after verification of inclusion criteria) : beginning of Product 2 administration; general medical examination; severity of diverticular disease symptoms; fecal sample for colonization study;

[0050] V2 - second study visit (6 weeks after inclusion) : completion of Product 2 administration, general medical examination; control of adverse reactions; fecal sample for colonization study; intensity of diverticular disease symptoms;

[0051] V3 - third study visit (9 weeks after inclusion and 3 weeks after last administration of the probiotic composition) : general medical examination; control of adverse reactions; intensity of diverticular disease symptoms; fecal sample for colonization study .

[0052] 3.2. The method for determination of patients' intestinal colonization with strains administered with Product 2 having the composition as disclosed in Table 2. Fecal samples sel f-collected by patients into test tubes filled with a buf fer to protect the genetic material using sampling spoons attached to the tube caps prior to visit VI , V2 , and V3 ( in duplicates ) . Metagenomic DNA isolated by column-based extraction, concentration measured by spectrophotometry, stored at -20 ° C until submitted to qPCR assay . The presence of each strain within the preparation veri fied by 2 reactions : 2 pairs of primers to detect species-speci fic and strain ( or a group of strains ) -speci f ic markers , the identity of known genomic fragments being >98 % (based on the BLAST analysis of the NCBI GenBank database content ) ; the result considered positive only in the case of both products being obtained (veri fication of softening temperature by comparison with a positive control reaction involving the genomic DNA of the desired strain as performed in parallel to the test reaction) . In addition, a positive control with universal 16S rRNA starters and a negative control without matrix were run for each assay

[0053] The results are shown in Figures 7 and 8 .

[0054] The presented results are indicative , in a consistent and statistically unambiguous manner, of a synergistic ef fect in relation to the tested parameters , said synergistic ef fect being impossible to obtain, on the basis of literature data, from independent administration of a source of butyric ions and probiotic bacteria . The results are characteri zed by high statistical signi ficance in relation to the results of the patients ' subj ective assessments , the clinical parameters , and the ability of bacteria to permanently coloni ze the large intestine . The technical problem solved by the invention leads to a clear and unexpected improvement in the condition of patients with SUDD The butyrate source can be used interchangeably with tributyrate , as the content of butyrate ions , not the source of their origin, is of importance here . The minimum content of butyrate ions is 101 mg per capsule . The minimum content of 5 . 2 x 109CFU is that of preferably 2-4 strains , including L . rhamnosus and L . plantarum, as best coloni zation has been observed for these parameters . The butyrate ions added to the composition comprise a prebiotic compound capable of supporting probiotic strains for ef fective intestinal coloni zation . In addition, butyrate has a well-documented role in the treatment of intestinal diseases owing to its trophic ef fect on human epithelial cells . Short-chain fatty acids ( SCFA) are formed by beneficial gut bacteria as the product of the metabolism of polysaccharides undigested by human enzymes within the digestive system . They are also the main energy substrate for epithelial cells of the intestinal mucosa . Increasing numbers of scienti fic reports indicating the importance of SCFA, especially butyric acid, in the treatment of many diseases of the colon, are being published ( Tan, 2014 ) . Butyric acid plays a signi ficant role in the metabolism and physiology of the lining of the gastrointestinal tract . It is rapidly absorbed and metaboli zed by epithelial cells along the entire digestive tract . Therefore , slow and controlled release throughout the small and large intestine is of great importance for proper therapeutic results . Hence , butyrate preparations are appropriately protected for administration in speci fic formulations ( enteral tablets / capsules , lipid carriers , polysaccharide carriers , etc ) . Prolonged administration of coated butyrate resulted in clinical improvement or remission in mild Crohn ' s disease ( Di Sabatino , 2005 ) . The researchers further noted that simple enteral formulations of butyrate fail to ensure optimum distribution in the lumen of the gastrointestinal tract as the release of the entire butyrate content occurs in an single burst , i . e . the butyrate is released upon the breakdown of the protective coating of the tablet or capsule at a rather random location, and subsequently nearly immediately and locally used without reaching the epithelial damage area .

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Claims

Claims1. Fixed-combination pharmaceutical composition of a butyrate compound and probiotic bacteria for use in the treatment and prevention of symptomatic uncomplicated diverticular disease2. Fixed-combination pharmaceutical composition for use according to claim 1 characterized in that the butyrate compound is sodium butyrate or tributyrin.

3. Fixed-combination pharmaceutical composition for use according to claim 1 or 2 characterized in that the daily dose of butyrate ions is 237 to 474 mg.

4. Fixed-combination pharmaceutical composition for use according to any of the claims from 1 to 3 characterized in that the daily dose of probiotic bacteria is 10.4 x 109CFU to 16.4 x 109CFU, preferably administered in two doses of 5.2 x 109to 8.2 x 109CFU per dose.

5. Fixed-combination pharmaceutical composition for use according to any of the claims from 1 to 4 characterized in that the probiotic bacteria are selected from the group comprising Lactobacillus rhamnosus , Lactobacillus plantarum, Lactobacillus reuterll , Bl fldobacterl um Inf antis .

6. Fixed-combination pharmaceutical composition for use according to any of the claims from 1 to 5 characterized in that it additionally contains probiotic yeast, preferably Saccharomyces cerevisiae var. boulardii, in the quantity of 4.0 x 109CFU per daily dose.. Fixed-combination pharmaceutical composition for use according to any of the claims from 1 to 6 characteri zed in that acylglycerols are selected from the group comprising mono- , di- and / or triacylglycerols of plant origin .8 . Fixed-combination pharmaceutical composition for use according to any of the claims from 1 to 7 characteri zed in that the butyrate compound is in the form of acylglycerol-coated microgranules .

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