DESC:PROBIOTIC FORMULATION FOR TREATMENT OF GASTROINTESTINAL DISORDERS

TECHNICAL FIELD
[001] The present subject matter generally relates to a probiotic formulation, more particularly the subject matter relates to a probiotic formulation for treatment of gastrointestinal disorders, even more particularly it relates to a probiotic formulation for treatment of gastrointestinal disorders, especially, constipation.
BACKGROUND OF THE INVENTION
[002] Constipation is the most common form of gastrointestinal disorders in humans. Almost one adult in five suffer from temporary or permanent constipation. It is a functional disorder in which, the adult is not able to empty his bowels normally and therefore needs external medications or laxatives, for softening the fecal material in the intestines and for easy expelling of the waste.
[003] Though laxatives are not very harmful for general human health, scientific studies have shown that they may cause imbalances in the gut micro biota, often resulting in loss of various useful intestinal microbial species, thereby disturbing normal intestinal functioning.
[004] Restoration of useful microbes is required for normal breakdown and absorption of nutrients in the intestine. One solution involves dietary supplementation with probiotic bacteria that are expected to lodge themselves in the intestine and provide beneficial effects to the individual. Probiotics are non-pathogenic and beneficial microbes which can be taken orally in form of tablets, powders or liquids. These microbes lodge themselves in the gut, colonize the gut environment and breakdown the food entering in the intestine, thereby aiding in digestion of the individual. Probiotics are required to be resistant to the highly acidic environment of the stomach and to reach the intestine without being destroyed.
[005] US patent application 2016/0192689 describes a functional food composition, made of probiotic and prebiotic components that are enclosed in a matrix. The food composition provides nutritive and health benefits for disease treatment and prevention, and health maintenance through the incorporation of certain biologically active compounds in the composition. US patent application 2016/0353774 describes a nutritional composition for pediatric subjects. The disclosure relates to nutritional compositions comprising a spore-forming probiotic suitable for administration to pediatric subjects. Additionally, the disclosure relates to methods of delivering nutritional compositions that contain probiotics to pediatric subjects for management of diarrhea, prevention or reduction of allergic skin conditions, and enhancement of the immune function. US patent application 2018/0042972 describes a composition made of a base substance, one or more prebiotics, and one or more probiotics, to promote GI health and enhancing immunity. Thus, there is a need to develop a probiotic formulation for safe and natural treatment of constipation.
OBJECTS
[006] Accordingly, the main object of the present invention is to provide a probiotic formulation for treatment of gastrointestinal disorders, especially, constipation and improve overall gastrointestinal health.
SUMMARY OF THE INVENTION
[007] To meet the above objects and to overcome the problems existing in the prior art, the present subject matter describes a probiotic formulation for treatment of gastrointestinal disorders, especially, constipation and improve overall gastrointestinal health.
[008] The probiotic formulation of the present subject matter is an oral suspension. The formulation contains a synthetic prebiotic component. Prebiotics are edible compounds that are nonabsorbable and nondigestible in gastro intestinal tract of humans. They are generally not affected by the acidic condition of the stomach and are not absorbed in the small intestine. Prebiotics are degraded in the gastro intestinal tract by the bacterial flora present therein. The prebiotic component described herein is a synthetic disaccharide lactulose. The amount of lactulose is 40 percent to 80 percent in the formulation. Prebiotics are known to breakdown in the gut of the host and provide a source of nutrition for probiotic bacteria. Prebiotics are also known to enhance the growth of the probiotic bacteria thereby helping in outnumbering the slower growing pathogenic organisms in the gut of the host.
[009] The present formulation also contains a probiotic component. Probiotics are viable microbes that lodge themselves in the gut of an animal and are beneficial to the host animal, by providing intestinal microbial balance. The probiotic microorganisms referred to herein are Bacillus coagulans Unique IS2 MTCC 5260.These microorganisms break down the prebiotic lactulose into compounds which destroy or outnumber the pathogenic bacteria of the gut.
[0010] The probiotic component described herein is in form of granules. The granules are composed of a mixture of a diluent and viable spores of the probiotic spore forming bacteria Bacillus coagulans Unique IS2 MTCC 5260. The effective amount of Bacillus coagulans Unique IS2 MTCC 5260 is 106 spores to 1012 spores per dosage of said formulation. The diluent is selected from a group consisting of stearic acid, microcrystalline cellulose, gelatine, magnesium stearate. The diluent described herein is microcrystalline cellulose. The amount of diluent is based on the requirement of strength of spores of the probiotic spore forming bacteria Bacillus coagulans Unique IS2 MTCC 5260 in the said oral suspension. The main role of the diluent is to serve as a bulking agent in pharmaceutical and nutraceutical preparations.
[0011] The spores of probiotic spore forming bacteria Bacillus coagulans Unique IS2 MTCC 5260 and the diluent are mixed and granulated to ensure smooth flow of the formulation. A granulating agent is used for formation of said granules. The granulating agent is selected from the group consisting of an acrylate, alginate, cellulose, cellulose derivatives, dextran, gelatine, dextrin, polyvinylpyrrolidone, and starch. The amount of granulating agent is 2 percent to 20 percent.
[0012] The granules of spores of probiotic spore forming bacteria Bacillus coagulans Unique IS2 MTCC 5260 and the diluent are subjected to microencapsulation for providing enteric protection to the said granules and also for extended release of the spores of probiotic bacteria into the gastrointestinal tract, because it is the main purpose of the invention that the bacteria should remain viable in the harsh acidic environment of the stomach and reach the gut. The microencapsulating agent is selected from the group consisting of acrylic resin, polyvinyl alcohol, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose, sodium carboxymethyl cellulose, ethyl cellulose, and acrylic resin. The amount of encapsulating agent is 10 percent to 30 percent.
[0013] The probiotic formulation further contains a flavouring agent selected from the group consisting of vanilla, mango, ripe banana and lemon. The amount of flavouring agent is 0.3 percent to 1.6 percent in the oral suspension.
[0014] The subject matter also describes a process for preparation of a probiotic formulation, comprising the steps of providing dry spores of Bacillus coagulans Unique IS2 MTCC 5260; blending said spores with a diluent to form a mixture; granulating said mixture with a granulating agent preferably a granulating liquid to form granules; microencapsulating said granules for enteric protection and extended release of said probiotic formulation; mixing the encapsulated granules with appropriate amount of a prebiotic substance wherein the prebiotic substance is preferably in a liquid form; adding a flavouring agent and adding appropriate amount of sterilised water to make an oral suspension.
[0015] The subject matter therefore describes a probiotic formulation for gastrointestinal disorders comprising a synthetic prebiotic compound and the said prebiotic compound is nonabsorbable and nondigestible in a gastro intestinal tract of humans, a plurality of microencapsulated granules comprising a mixture of a diluent and viable spores of a probiotic spore forming bacteria capable of degrading said synthetic prebiotic compound and a flavouring agent wherein said formulation is in form of oral liquid suspension, sachets, powder/gel for reconstitution.
[0016] The subject matter also describes a method of providing a probiotic formulation for treatment of gastrointestinal disorders. The method includes oral administration of at least a therapeutically effective dose of a probiotic formulation a probiotic spore forming Bacillus coagulans Unique IS2 MTCC 5260, a prebiotic compound, a diluent and a flavouring agent. The effective dose is in form of oral liquid suspension, sachets, powder/gel for reconstitution. The formulation may be continued till the signs of constipation are alleviated.
DETAILED DESCRIPTION OF THE INVENTION
[0017] Accordingly, it is an object of the present subject matter to describe a probiotic formulation comprising a prebiotic, a probiotic and a flavouring agent, said formulation being in the form of an oral suspension.
[0018] In an object, the prebiotic is a synthetic disaccharide selected from lactulose, fructo oligosaccharide, inulin, and lactilol.
[0019] In another object, the synthetic disaccharide is nonabsorbable and nondigestible in a gastro intestinal tract of humans.
[0020] In one more object, the amount of synthetic disaccharide is 40 percent to 80 percent.
[0021] In yet another object, the probiotic is composed of a plurality of microencapsulated granules and the granules are composed of a mixture of a diluent and viable spores of a probiotic spore forming bacteria capable of specifically degrading the synthetic prebiotic compound.
[0022] In one more object of the present subject matter, the spore forming bacteria belong to Bacillus species, specifically, Bacillus coagulans Unique IS2 MTCC 5260 and the effective amount of Bacillus coagulans Unique IS2 MTCC 5260 is 106 spores to 1012 spores per dosage of said formulation,
[0023] In yet another object, the diluent is selected from a group consisting of stearic acid, microcrystalline cellulose, gelatin, magnesium stearate, and the amount of diluent is based on the requirement of strength of spores of the probiotic spore forming bacteria Bacillus coagulans Unique IS2 MTCC 5260 in the said oral suspension.
[0024] In one more object, a granulating agent is used for formation of said granules mainly because granulation allows a smooth flow and protection of viability of the said probiotic formulation.
[0025] In yet another object, the granulating agent is selected from the group consisting of an acrylate, alginate, cellulose, cellulose derivatives, dextran, gelatine, dextrin, polyvinylpyrrolidone and starch and is mixed with the mixture of diluent and spores of Bacillus coagulans Unique IS2 MTCC 5260 in an amount of 2 percent to 20 percent.
[0026] In an object, microencapsulation is done with a purpose of providing enteric protection to the said granules and for extended release of the probiotic bacteria into the gastrointestinal tract of humans or an animal.
[0027] In one more object a microencapsulating agent is selected from the group consisting of acrylic resin, polyvinyl alcohol, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose, sodium carboxymethyl cellulose, ethyl cellulose, preferably hydroxypropyl methylcellulose (HPMC) and it is used in an amount of 10 percent to 30 percent.
[0028] In an object, there is a flavouring agent added to the probiotic formulation and the flavouring agent is selected from the group consisting of vanilla, mango, banana and lemon.
[0029] In one more object, the amount of flavouring agent is 0.3 percent to 1.6 percent preferably 0.5 percent to 1 percent in the oral suspension.
[0030] In another object, a process for preparation of a probiotic formulation, said process comprising providing dry spores of Bacillus coagulans Unique IS2 MTCC 5260; blending said spores with a diluent to form a mixture; granulating said mixture with a granulating agent preferably a granulating liquid to form granules; encapsulating said granules for enteric protection and extended release of said probiotic formulation; mixing the encapsulated granules with appropriate amount of a prebiotic substance and adding a flavouring agent.
[0031] In an object, the probiotic formulation for gastrointestinal disorders is made of a synthetic prebiotic compound and viable spores of a probiotic spore forming bacteria capable of degrading said synthetic prebiotic compound and a flavouring agent and the said formulation is in form of oral liquid suspension, sachets, powder/gel for reconstitution.
[0032] In another object, a method of providing a probiotic formulation for treatment of gastrointestinal disorders is described. The method includes oral administration of at least a therapeutically effective dose of a probiotic formulation a probiotic spore forming Bacillus coagulans Unique IS2 MTCC 5260, a prebiotic compound a diluent and a flavouring agent. The effective dose is in form of oral liquid suspension, sachets, powder/gel for reconstitution.
DETAILED DESCRIPTION OF THE SUBJECT MATTER WITH RESPECT TO EXAMPLES
[0033] Before the present subject matter is described in further detail, it is to be understood that the subject matter is not limited to the particular embodiments described, and as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims. It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items unless the context clearly dictates otherwise.
[0034] EXAMPLE 1
[0035] A double-blind, randomized, placebo-controlled, parallel group multicentric out-patient study on human volunteers was conducted. The study was conducted in compliance with the code of conduct for research involving human volunteers as issued by the International Conference on Harmonization Good Clinical Practice (ICH-GCP), Indian Council of Medical Research guidelines (ICMR; ethical guidelines for biomedical research on human subjects) and the principles of the Declaration of Helsinki. The study was initiated after obtaining informed consent approved by the ethical committees of the concerned hospitals and the trial was registered prospectively with the clinical trial registry of India (CTRI/2017/11/010539).
[0036] The ITT (intention-to-treat) population consisted of randomized subjects who received one dose of either B. coagulans Unique IS2 (2 billion CFU) capsule (n = 50) or placebo (n = 50) with a post-baseline efficacy assessment. The strength (2 billion CFU) of B. coagulans in the capsule was confirmed by plating a serially diluted sample on GYE (glucose yeast extract) agar. The PP (per protocol) population consisted of subjects, both B. coagulans Unique IS2 (n = 50) and placebo (n = 50) having completed the study without any major protocol deviation. Of the 100 patients, 50 received probiotic, B. coagulans Unique IS2 capsules (2 × 109 CFU) and 50 received placebo daily for up to 4 weeks. The mean age of total patients was 43.92 ± 11.74 years (range, 18–64 years); 59 (59%) patients were males and 41 (41%) were females. The mean age of patients who received B. coagulans Unique IS2 was 42.54 ± 12.16 years (range, 18–64 years) and placebo was 45.30 ± 11.33 years (range, 24–64 years). Thirty-four (68%) patients were males and 16 (32%) patients were females in B. coagulans Unique IS2 group, whereas 25 (50%) patients were males and 25 (50%) were females in placebo.
[0037] Inclusion criteria: Subjects of either sex between 18 and 65 years of age with diagnosis as functional constipation according to Rome Criteria III; should not have/had any major illnesses; willing to give written informed consent and follow study procedures.
[0038] Exclusion criteria: Subjects who had undergone major abdominal surgery; documented history of slow colonic transit; presence of any concomitant diseases such as organic GI diseases and/or lactose and gluten intolerance; medical or psychiatric illness; clinical features suggestive of alarming symptoms (rectal bleeding, weight loss etc.); family history of peptic ulcer, colorectal cancer or IBD, abnormal laboratory data or thyroid function; participation in any clinical trial or usage of any investigational product in the past 90 days; known or expected hypersensitivity to any of the active substances or excipients; used any probiotic formulation in the past 30 days; pregnant or lactating; allergic or atopic to any of the ingredients of the study medication.
[0039] Randomization
[0040] After having obtained signed, written informed consent, subjects underwent a screening examination. Subjects complying with inclusion/exclusion criteria were enrolled and randomized by block randomization to one of the two treatment arms. Based on SAS 9.4 randomization, numbers for two treatment groups were generated. Randomization was conducted using opaque sealed envelopes that were indistinguishable between groups in order for the investigators also to be blinded to the treatment. Each envelope had the assignment of the patient (probiotic or placebo treatment) with 50 envelopes for each group. The sealed envelopes were provided to the clinical site. The investigators assigned investigational products to patients based on the randomization numbers. Both groups were characteristically similar pertaining to age, sex and weight of the patient.
[0041] Study Follow-up Visits and Treatments
[0042] The duration of treatment was for a period of 28 days with a follow up until the end of the treatment. B. coagulans Unique IS2 or placebo capsules were administered once daily for up to 28 days. The secondary efficacy endpoints were changes from baseline to end of treatment in stool frequency, stool consistency (as assessed by Bristol stool form scale), difficulty in degree of defecation, sensation of incomplete evacuation (CCS scale, constipation scoring system), defecation pain (CCS scale) and abdominal pain (CCS scale). Safety endpoints included the incidences of treatment-emergent adverse events, abnormal vital signs and clinically significant changes from baseline in physical examination.
[0043] Statistical Methods
[0044] All efficiency analyses were performed on ITT population which remained the same as PP population due to no major protocol deviations. Primary endpoint qualitative data was defined as number and percentages, and the data was compared using unpaired t test at 5% level of significance between groups. The secondary endpoint data was also interpreted as descriptive data for scores as n, mean, median, standard deviation and range (minimum and maximum). Data was analyzed using unpaired t test at 5% level of significance between groups.
[0045] Evaluation of Results
[0046] The main criteria in the evaluation of results was to see if there was an improvement in the primary efficacy parameter, i.e. treatment success based on stool frequency (three or more spontaneous stools per week). As the study was a randomized, double-blind study, the differences observed between the two groups (probiotic and placebo) could be ascribed solely to the effect of the treatment as bias was removed. To arrive at the required sample size for the study (50 subjects/arm), sample size calculations were used to arrive at the sample size to identify a significant result in this primary outcome measure.

[0047] Results
[0048] Primary Efficacy
[0049] Treatment Success Based on Stool Frequency (Defined as Three or More spontaneous Stools per Week)
[0050] In patients receiving B. coagulans Unique IS2 capsules, the bowel frequency was 0.90 ± 0.73 (week 1), 1.66 ± 1.81 (week 2), 4.16 ± 1.98 (week 3) and 5.98 ± 1.57 (week 4) spontaneous stools per week. In the placebo group, bowel frequency was 0.94 ± 0.86 (week 1), 1.62 ± 1.78 (week 2), 2.34 ± 1.31 (week 3) and 3.12 ± 1.18 (week 4) spontaneous stools per week. At week 3 and week 4, there was a significant improvement (p < 0.001) in mean frequency of spontaneous stools per week in patients receiving B. coagulans Unique IS2 capsules as compared to placebo.
[0051] Secondary Efficacy
[0052] Stool Consistency
[0053] Stool consistency was assessed by Bristol stool form scale. The stool consistency improved significantly in the probiotic treated group of the functionally constipated subjects from the third week onwards. There was thus an increase in the percentage of patients with normal stool in the B. coagulans uniqueIS2 treated group as compared to placebo group.
[0054] Difficulty in Degree of Defecation and Sensation of Incomplete Evacuation (CSS Scale)
[0055] The sensation of incomplete evacuation was assessed through constipation scoring system (CSS) scale. The CSS score ‘completeness: feeling incomplete evacuation’ score was a scale of 0–4 with higher CSS scores indicating more severe constipation (0, never; 1, rarely; 2, sometimes; 3, usually; and 4, always). In patients receiving B. coagulans Unique IS2 capsules, there was a significant decrease as compared to placebo in the feeling of incomplete evacuation as assessed by constipation scoring system. By visit 3, B. coagulans Unique IS2 treated group had a mean score of 1.32 ± 0.51 as compared to placebo which was 1.62 ± 0.73 (p = 0.019), and by visit 4, in the B. coagulans Unique IS2 treated group, the score had further dropped to 0.88 ± 0.39 as compared to placebo which was 1.04 ± 0.73 (p = 0.034).
[0056] Defecation Pain (CSS Scale)
[0057] The severity of constipation symptom ‘difficulty: painful evacuation effort’ was assessed through CSS scale. The CSS ‘difficulty: painful evacuation effort’ score was a scale of 0–4 with higher CSS scores indicating more severe pain (0, never; 1, rarely; 2, sometimes; 3, usually; and 4, always). By visit 3, B. coagulans Unique IS2 treated group had a mean score of 1.16 ± 0.58 as compared to placebo which was 1.50 ± 0.84 (p = 0.021), and by visit 4, in B. coagulans Unique IS2 treated group, the score had further dropped to 0.66 ± 0.52 as compared to placebo which was 0.98 ± 0.62 (p = 0.006).
[0058] Abdominal Pain (CSS Scale)
[0059] The severity of constipation symptom ‘pain: abdominal pain’ was assessed through CSS scale. The CSS ‘pain: abdominal pain’ score was a scale of 0–4 with higher CSS scores indicating more severe pain (0, never; 1, rarely; 2, sometimes; 3, usually; and 4, always). By visit 3, B. coagulans Unique IS2 treated group had a mean score of 0.94 ± 0.68 as compared to placebo which was 1.1 ± 0.84 (p = 0.038) and by visit 4, in B. coagulans Unique IS2 treated group, the score had further dropped to 0.38 ± 0.49 as compared to placebo which was 0.8 ± 0.81 (p = 0.002).
[0060] Adverse Events
[0061] No serious adverse events were observed during the trial. There were three patients from the placebo group who had reported mild adverse events during the study. One patient had mild fever (pyrexia) and two patients had reported headache from placebo group which the investigator had considered as not related to study drug.
[0062] The vital parameters in both groups remained within normal and acceptable clinical range throughout the study duration. In the present study, adults with functional constipation who received B. coagulans Unique IS2 capsules had improved bowel movements as compared to group fed with placebo. The present study suggests that B. coagulans Unique IS2, a clinically proven and safe probiotic, can be used in the treatment of constipation.
[0063] While the invention has been described with reference to preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, the description is not intended to limit the scope of the invention to the particular forms set forth, but on the contrary, it is intended to cover such alternatives, modifications, and equivalents as may be included within the spirit and scope of the invention as indicated by the following appended claims.

,CLAIMS:We claim:
1. A probiotic formulation comprising:
a prebiotic component, wherein said prebiotic component is a synthetic nonabsorbable disaccharide, nondigestible in a gastro intestinal tract of humans;
a probiotic component comprising a plurality of microencapsulated granules said granules further comprising a mixture of a diluent and viable spores of a probiotic spore forming Bacillus coagulans capable of degrading said synthetic prebiotic compound, and
a flavouring agent.
2. The probiotic formulation of claim 1, wherein the formulation is an oral suspension.
3. The probiotic formulation of claim 1, wherein the prebiotic component is selected from a group comprising lactulose, inulin, fructo oligosaccharide and lactilol, preferably synthetic disaccharide lactulose.
4. The probiotic formulation of claims 2 and 3, wherein the amount of lactulose is 40 percent to 80 percent in the said formulation.
5. The probiotic formulation of claim 1, wherein the probiotic spore forming bacteria are Bacillus coagulans Unique IS2 MTCC 5260.
6. The probiotic formulation of claim 5, wherein the effective amount of Bacillus coagulans Unique IS2 MTCC 5260 is 106 spores to 1012 spores per unit dosage of said formulation.
7. The probiotic formulation of claim 1, wherein the diluent is selected from a group consisting of stearic acid, microcrystalline cellulose, gelatine, magnesium stearate, preferably microcrystalline cellulose.
8. The probiotic formulation of claim 1, wherein a granulating agent is used for formation of said granules wherein said granulating agent supports a smooth flow and protects viability of spores of the said probiotic formulation.
9. The probiotic formulation of claim 8, wherein the granulating agent is selected from the group consisting of an acrylate, alginate, cellulose, cellulose derivatives, dextran, gelatine, dextrin, polyvinylpyrrolidone, and starch, preferably polyvinylpyrrolidone.
10. The probiotic formulation of claim 1, wherein microencapsulation is used to provide enteric protection to the said granules.
11. The probiotic formulation of claim 1, wherein microencapsulation is used for extended release of the probiotic bacteria into the gastrointestinal tract.
12. The probiotic formulation of claims 10 and 11, wherein the microencapsulating agent is selected from the group consisting of acrylic resin, polyvinyl alcohol, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose, sodium carboxymethyl cellulose and ethyl cellulose.
13. The probiotic formulation of claim 1, wherein the flavouring agent is selected from the group consisting of vanilla, mango, banana and lemon.
14. The probiotic formulation of claims 2 and 13, wherein the amount of flavouring agent is 0.3 percent to 1.6 percent in the said formulation.
15. A process for preparation of a probiotic formulation, said process comprising:
providing dry spores of Bacillus coagulans Unique IS2 MTCC 5260;
blending said spores with a diluent to form a mixture;
granulating said mixture with a granulating agent preferably a granulating liquid to form granules;
microencapsulating said granules for enteric protection and extended release of said probiotic formulation;
mixing the microencapsulated granules with appropriate amount of a prebiotic substance;
adding a flavouring agent;
adding appropriate amount of sterilised water to make an oral suspension.
16. The process of claim 9, wherein the amount of granulating agent is 2 percent to 20 percent.
17. The process of claim 12, wherein the amount of microencapsulating agent is 10 percent to 30 percent.
18. A probiotic formulation for gastrointestinal disorders comprising a synthetic prebiotic compound wherein said prebiotic compound is nonabsorbable and nondigestible in a gastro intestinal tract of humans, a plurality of encapsulated granules comprising a mixture of a diluent and viable spores of a probiotic spore forming bacteria capable of degrading said synthetic prebiotic compound and a flavouring agent wherein said formulation is in form of oral liquid suspension, sachets, powder/gel for reconstitution.
19. A method of providing a probiotic formulation for treatment of gastrointestinal disorders, said method comprising administering orally at least a therapeutically effective dose of a probiotic formulation, said formulation comprising a probiotic spore forming Bacillus coagulans Unique IS2 MTCC 5260, a prebiotic compound lactulose, a diluent and a flavouring agent.