NON-VIABLE BIFIDOBACTERIUM BIFIDUM BACTERIA AND USES THEREOF

Abstract

The present invention relates to non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514, or one or more fragments thereof, for use in therapy, particularly for use in treating a gastrointestinal disorder, such as irritable bowel syndrome. Furthermore, the present invention relates to a composition comprising, as an active ingredient, said non-viable bacteria for use in therapy, particularly for use in treating a gastrointestinal disorder. Also, the present invention relates to a method of preparing non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514, or one or more fragments thereof, and to bacteria obtained by the inventive method for use in therapy, particularly for use in treating a gastrointestinal disorder.

Claims

1-3. (canceled)

4. A composition comprising, as an active ingredient, non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514, or one or more fragments thereof.

5. (canceled)

6. The composition of claim 4, wherein the composition is a pharmaceutical composition or a food composition.

7. The composition of claim 4, wherein the composition comprises, as an active ingredient, non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514.

8. A method of preparing non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514, or one or more fragments thereof, the method comprising: (a) providing bacteria of the Bifidobacterium bifidum strain deposited under deposit No. DSM 24514; and (b) inactivating the bacteria provided in step (a) to obtain non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001.

9. The method of claim 8, wherein the bacteria are inactivated in step (b) by subjecting the bacteria to heat, pressure, sonication, irradiation, drying, pulsed electric field (PEF), supercritical CO.sub.2 and/or a pH change.

10.-11. (canceled)

12. A method of treating a gastrointestinal disorder, the method comprising administering non-viable bacteria, or one or more fragments thereof, obtainable by the method of claim 8 to a subject in need thereof.

13. (canceled)

14. A method of treating a gastrointestinal disorder, the method comprising administering non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514, or one or more fragments thereof, to a subject in need thereof.

15. (canceled)

16. The method of claim 14, wherein the gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, pouchitis, post infection colitis, diarrhoea, constipation, dyspepsia and/or associated dyspeptic symptoms, gastroparesis, and intestinal pseudo-obstruction.

17. The method of claim 14, wherein the non-viable bacteria are administered orally in a daily amount of at least about 10.sup.2 non-viable cells.

18. The method of claim 14, wherein the non-viable bacteria, or the one or more fragments thereof, are administered to a human subject.

19. The method of claim 14, wherein the non-viable bacteria, or the one or more fragments thereof, are administered to an immuno-compromised subject.

20. The method of claim 14, wherein the method comprises administering non-viable bacteria of the Bifidobacterium bifidum strain SYN-HI-001 deposited under deposit No. DSM 24514 to the subject.

Description

[0082] The figures show:

[0083] FIG. 1: Study design. A 12-week study including a run-in, treatment and wash-out period with five physician visits.

[0084] FIG. 2: Flowchart of the patients through the study. Verum=heat-inactivated B. bifidum SYN-HI-001.

[0085] FIG. 3: Composite responders (adequate relief responder combined with pain responder) during treatment in “intention-to-treat” (ITT) and “per-protocol” (PP) population.

[0086] FIG. 4: Course of composite responders (adequate relief responder combined with pain responder) during both treatment intervals. Significant more responders in B. bifidum SYN-HI-001 during both treatment intervals.

[0087] FIG. 5: Adequate relief responders during treatment.

[0088] FIG. 6: Comparison of effects of B. bifidum SYN-HI-001 and placebo on symptom relief (recorded on a 1-7 scale) on a weekly basis. Significant improvement in the Bifidobacteria group vs. placebo group from the second week of treatment till the end of treatment.

[0089] FIG. 7: Patients with ≥50% IBS-SSS improvement at the end of treatment in “intention to treat” (ITT) and “per protocol” (PP) population.

[0090] FIG. 8: Significant reduction of IB-SSS sumscore (0-500) by B. bifidum SYN-HI-001 in comparison to placebo on mean score changes from baseline to end of treatment in “intention-to-treat” (ITT) and “per-protocol” (PP) population.

[0091] FIG. 9: Significant reduction of IBS symptoms assessed on a daily basis by B. bifidum SYN-HI-001 in comparison to placebo on mean score changes from baseline at the end of treatment (“intention-to-treat”, ITT). Composite score 1-4=arithmetic mean of four individual symptom scores (SGA, pain, distension/bloating, urgency).

[0092] FIG. 10: Significant reduction of IBS symptoms assessed on a weekly basis by B. bifidum SYN-HI-001 in comparison to placebo on mean score changes from baseline at the end of treatment (“intention-to-treat”, ITT).

[0093] FIG. 11: Significant increase of SF-12 sum and mental health sum by B. bifidum SYN-HI-001 in comparison to placebo on mean score changes from baseline to end of treatment (“intention-to-treat”, ITT).

[0094] The following examples illustrate the invention:

EXAMPLE 1: A DOUBLE-BLIND, MULTI-CENTRE, RANDOMIZED AND PLACEBO-CONTROLLED STUDY ASSESSING THE EFFICACY OF HEAT-INACTIVATED B. BIFIDUM SYN-HI-001 IN PATIENTS WITH IRRITABLE BOWEL SYNDROME

[0095] 1.1 Study Population

[0096] Patients were recruited from principal investigators and by advertisement. The study protocol has been presented to the Ethics Committee of the Physicians Chamber of Hamburg for advisory opinion. Subjects aged between 18 and 65 years with IBS according to the Rome III criteria have been included. Excluded were individuals with inflammatory organic gastrointestinal diseases, systemic diseases, cancer, autoimmune diseases, diabetes, known lactose intolerance or immunodeficiency, abdominal surgery (except appendectomy, hernia surgery, cholecystectomy or caesarean section), no negative diagnostic result of sigmoidoscopy or colonoscopy within the last five years for patients older than 55 years, diagnosed hyperthyroidism, use of antipsychotics for at least 3 months or systemic corticosteroids during at least one month prior to study start, major psychiatric disorder, celiac disease or pregnancy.

[0097] 1.2 Study Design

[0098] The study was performed as a prospective, multi-centre, randomized, double-blind, placebo-controlled, two-arm interventional study. Throughout the study, patients recorded their global IBS symptoms as well as individual IBS symptoms daily using a patient diary. Additionally, patients have been questioned at a physician site for global and individual symptoms (visit 2-5) and quality of life (visit 2-4). Physician visits were conducted at screening, after two weeks run-in phase (randomization), after 4 weeks treatment (control visit), after 8 weeks (end of treatment) and after further two weeks wash-out phase (end of study) (FIG. 1).

[0099] After patients had given their written informed consent, they qualified for the screening examination at visit 1 (day 0), which included a complete medical history and physical examination. A blood sample was taken for analysis in a central laboratory, including a pregnancy test for women. Patients were instructed to maintain their eating and life style habits throughout the study. A patient diary was handed out.

[0100] At the second visit (day 14), diaries were reviewed by the physician. Patients who had recorded at least a pain score of ≥4 on an 11-point numerical rating scale (NRS) on two days during the run-in phase and who fulfilled all further inclusion criteria and did not violate any exclusion criteria were randomized 1:1 to receive either heat-inactivated B. bifidum SYN-HI-001 or placebo. Patients were allocated to the treatment groups according to a computer-generated blocked randomization list with a block size of 4. The block size was not communicated to the investigators and the allocation was blinded to both patients and site staff. During the 8 weeks intervention period, patients consumed either two capsules comprising heat-inactivated bacteria daily or an identical appearing placebo.

[0101] In the middle of the treatment phase (visit 3, day 42), investigators discussed the course of the disease with the patient and the first part of the treatment diary was collected and reviewed. At the end of the treatment phase (visit 4, day 70), investigators reviewed and collected the remaining treatment diary.

[0102] After the treatment-free wash-out phase (visit 5, day 84), investigators collected the wash-out diary as well as unused study product and empty cans in order to confirm compliance. Furthermore, a complete physical examination was conducted and a blood sample was taken. Bisacodyl and loperamide were allowed as rescue medication. Probiotics or other medications that could influence the efficacy of the study product were not allowed.

[0103] 1.3 Study Product Preparation

[0104] Viable cells of the Bifidobacterium bifidum strain deposited under deposit No. DSM 24514 were grown in a protein-rich liquid growth medium, heat-inactivated in a fermentation vessel, centrifuged and subsequently freeze-dried, milled and sieved. Furthermore, the dry powder inactivated bacteria were mixed with an excipient and filled into uncoated cellulose capsules in an amount of approx. 0.5×10.sup.9 cells (non-viable bacteria of Bifidobacterium bifidum strain SYN-HI-001). Placebo capsules appeared identical and contained maltodextrin.

[0105] 1.4 Endpoint Definitions

[0106] The prospectively defined primary efficacy variable was as a combination of ≥30% improvement of pain on an 11-point NRS and achievement of one of the best 3 relief categories on a 7-point Likert scale with these both criteria being fulfilled in at least 4 out of 8 weeks of treatment to qualify as a treatment responder. Patients were asked to answer the daily question “If you had experienced abdominal pain during the last 24 hours, how would you rate this pain?” Possible answers ranged from 0 (no pain) to 10 (strongest imaginable pain).

[0107] Relief of symptoms was captured weekly on a 7-point Likert scale (Global Assessment of Improvement Scale). This scale was assessed every week during the 8-weeks treatments period in the patient diary. Patients were asked to answer the weekly question “Compared to the way you usually felt before taking the study medication how would you rate your relief of symptoms (abdominal pain/discomfort, bowel habits and other IBS-symptoms) during the last 7 days?” Possible answers ranged from 1 (very much relieved), 2 (considerably relieved), 3 (somewhat relieved), 4 (unchanged), 5 (somewhat worse), 6 (considerable worse) to 7 (very much worse).

[0108] Secondary efficacy variables included the “Subject global assessment of symptoms (SGA)” and individual symptoms of IBS alone, such as “abdominal pain”, “distension/bloating” and “urgency”, recorded on the same 7-point Likert scale (abdominal pain was assessed on the 11-point NRS as described above). The individual symptom scores were additionally combined into a composite symptom score as the arithmetic mean of SGA and the three individual symptom scores. Furthermore, the reduced and/or increased number of bowel movements, stool form (assessed via the Bristol Stool Form Scale), feeling of incomplete bowel evacuation and intake of other medication were reported daily in the diary and evaluated.

[0109] At the beginning, middle and end of treatment as well as at the end of the study (visits 2-5), physicians questioned the patients regarding their severity of IBS symptoms via the IBS-severity scoring system (IBS-SSS). The score is based on 5 visual analogue scales (VAS) ranging from 0 to 100 and contains for the past 10 days the severity of the symptoms “abdominal pain severity”, “abdominal pain frequency” (number of days with pain during the last 10 days), “abdominal bloating severity”, “bowel movement satisfaction” and “interference of IBS with daily activities”. Health related quality of life was assessed by the use of the SF-12 questionnaire prior to treatment, at the middle of the treatment and end of treatment (visit 2, 3 and 4).

[0110] Adverse events were recorded throughout the study and the global assessment of tolerability of the study treatment has been questioned at physician visit 3, 4 and 5. Tolerability was assessed on a 5-point scale with the question “If you consider the side effects of the study treatment, how would you rate the overall tolerability?” Possible answers ranged from 1 (very good), 2 (good), 3 (fair), 4 (not satisfactory) to 5 (bad). Vital signs were checked at every physician visit and laboratory values were examined at the screening visit and the end of study.

[0111] 1.5 Statistical Methods

[0112] 1.5.1 Sample Size Calculation

[0113] Sample size calculation was based on both estimated responses of placebo and treatment group for the primary endpoint and estimated differences between the IBS subgroups for main symptom scores. With 80% power, a sample size of 350 evaluable patients was calculated. With an estimated drop-out rate of 15-20% after randomization, 412 randomized patients were planned and 507 were recruited to account for possible withdrawals prior to study start.

[0114] 1.5.2 Statistical Analysis

[0115] The primary objective of this study was to prove the hypothesis that the combined responder rate of adequate relief responder 3 and pain responder is significantly larger in the Bifidobacterium group compared to the placebo group. The primary endpoint was defined as a combination of ≥30% improvement of pain on an 11-point NRS (pain responder) and achievement of one of the best 3 relief categories on a 7-point Likert scale (adequate relief responder 3) and wherein these two criteria had to be fulfilled in at least 4 out of 8 weeks of treatment in order to qualify as a treatment responder. The Cochrane-Mantel-Haenszel test stratified by study center was used for the comparison of treatment arms and P<0.05 was considered as statistically significant.

[0116] The primary analysis was based on the intent-to-treat population where all successfully randomized patients were included. Weeks with missing data were automatically counted as no response. Patients who only provided baseline entries for the primary efficacy variable were considered as “non-responders” in the evaluation of the primary efficacy criterion. An additional per-protocol-analysis was performed for supportive purposes.

[0117] Secondary endpoints were analyzed descriptively based on available data. To detect treatment differences the Wilcoxon rank sum test was applied for continuous variables and the Fishers exact test for binary variables. All p values are two-sided. Secondary efficacy variables included response based on a 50% rule of symptom relief during treatment (at least improvement in 4 out of 8 weeks within the treatment period and improvement defined as at least one point reduction from baseline).

[0118] Adverse events (AE) were coded by using MedDRA using the current version at the time of study start which is used throughout the study. Coding was based on the German version and the corresponding English version was used for the final analysis. AEs were tabulated based on Preferred Terms (PT) of the MedDRA dictionary. Tabulations will include affected patients and absolute and relative frequencies of events.

[0119] All statistical analyses were performed using SAS version 9.5 for windows, SAS Institute Inc., Cary, N.C., USA.

[0120] 1.6 Results

[0121] 1.6.1 Subjects

[0122] From the 507 patients screened, 443 patients were successfully randomized to receive either placebo (n=222) or heat-inactivated B. bifidum SYN-HI-001 (n=221). All randomized patients were analyzed for intent-to-treat (ITT) and adverse events (n=443). A total of 377 patients (187 placebo and 190 B. bifidum SYN-HI-001) were analyzed as per-protocol (FIG. 2).

[0123] 1.6.2 Baseline Characteristics

[0124] Baseline characteristics and demographics were well balanced between the two treatment groups. 24.2% were classified as constipation-predominant IBS (IBS-C), 40.0% as diarrhoea-predominant IBS (IBS-D), 7.7% as mixed IBS (IBS-M) and 28.2% as unsubtyped IBS (IBS-U) with no significant differences between the Bifidobacteria and the placebo group.

[0125] On average, patients were 41.3 years, with 69.3% females, a mean height of 171.9 cm, mean weight of 73.0 kg and a mean BMI of 24.6, with no significant differences between the two treatment groups (Table 1).

TABLE-US-00001 TABLE 1 Demographic characteristics of the “intention-to-treat” (ITT) population B. bifidum SYN-HI-001 (N = 221) Placebo (N = 222) N (%) or mean ± s.d. N (%) or mean ± s.d. Age 40.1 ± 12.8 42.7 ± 13.8 Female Sex 155 (70.1) 152 (68.5) Height (cm) 172.4 ± 8.9  171.3 ± 9.1  Weight (kg) 73.2 ± 7.7  72.8 ± 16.6 BMI 24.5 ± 5.3  24.7 ± 5.0  IBS-type IBS-C  54 (24.4)  53 (23.9) IBS-D  95 (43.0)  82 (36.9) IBS-M 14 (6.3) 20 (9.0) IBS-U  58 (26.2)  67 (30.2)

[0126] 1.6.3 Primary Endpoint: Composite Responder (Adequate Relief Responder 3 Combined with Pain Responder)

[0127] The primary endpoint was the composite responder, defined as a combination of 30% improvement of pain and achievement of one of the best 3 relief categories in at least 4 out of the 8 treatment weeks (50%-rule). Based on this definition, composite response was achieved in 33.5% of patients in the Bifidobacterium group (74/221) compared to 19.4% of patients in the placebo group (43/222). This is equivalent to a 1.7-fold higher success rate (95% CI: 1.3-2.4) with the study product. The Cochrane-Mantel-Haenszel test stratified by study center proved a statistical significant difference (P=0.00071). The positive result was confirmed by the per-protocol analysis. Patients treated with heat-inactivated B. bifidum SYN-HI-001 achieved 36.8% composite response compared to 19.8% when treated with placebo (P=0.0004 with the Mantel-Haenszel Test). These results are highly significant and indicate clear superiority of heat-inactivated B. bifidum SYN-HI-001 over placebo (FIG. 3). Analysis of the course of the composite responder during the two treatment intervals (week 1-4 and week 5-8) showed that the treatment with B. bifidum SYN-HI-001 was significantly superior to placebo during both the first and the second treatment interval. During weeks 1-4 31.7% of the Bifidobacteria vs. 19.8% of the placebo group (P=0.0047) and during weeks 5-8 39.4% of the Bifidobacteria vs. 29.7% (P=0.0361) fulfilled the response criteria in 50% of the time (FIG. 4).

[0128] 1.6.4 Secondary Endpoints

[0129] 1.6.4.1 Other Combined Responders and Adequate Relief/Symptom Relief

[0130] Secondary endpoints included a “stricter” definition of composite response: combination of 30% improvement of pain and achievement of one of the best 2 relief categories in at least 4 out of the 8 treatment weeks. Even with this “stricter” definition of adequate relief responders, this composite response was significantly higher in the Bifidobacteria group (15.8%) compared to placebo (7.7%) (P=0.0079), with the result being confirmed in the per-protocol analysis (B. bifidum SYN-HI-001: 17.9% vs. placebo: 7.5%; P=0.0031).

[0131] Analysis of the adequate relief responder alone (and not combined with another responder) results in a significant benefit for the treatment with B. bifidum SYN-HI-001. Adequate relief 3 response rate (best 3 relief categories in at least 4 out of the 8 treatment weeks) was 60.18% in the Bifidobacteria group (i.e. the verum group) and only 44.14% in the placebo group (P=0.0009). Also, adequate relief 2 response rate (best 2 relief categories in at least 4 out of the 8 treatment weeks) was significantly higher in the verum group (20.36%) compared to placebo (11.26%) (P=0.0093) (FIG. 5).

[0132] Another secondary endpoint was an improvement in symptom relief (a reduction of the mean score), which was assessed every week during the 8-weeks treatment in the patient diary. The evaluation of the symptom relief on a weekly basis showed a significant benefit for patients within the Bifidobacteria group for every single week starting from the second week of treatment (week 2) until the end of treatment (week 8). At the end of treatment, the mean score in symptom relief was 3.08 in the verum group vs. 3.44 in the placebo group (P=0.006) (FIG. 6).

[0133] 1.6.4.2 Irritable Bowel Syndrome-Severity Scoring System (IBS-SSS)

[0134] IBS-SSS was assessed at baseline (visit 2), at the control visit after 4 weeks of treatment (visit 3), after the end of treatment at week 8 (visit 4) and at the end of the study at week 10 (visit 5) with a sum score ranging from 0-500. At the end of treatment (week 8), the IBS-SSS could be improved ≥50% in significantly more patients of the verum group (41.2%) as compared to patients in the placebo group (28.8%) (P=0.0072). The result was confirmed in the per-protocol analysis (B. bifidum SYN-HI-001: 37.4% vs. placebo: 25.1%; P=0.0109) (FIG. 7).

[0135] Regarding the reduction of IBS-SSS sum score from baseline to the end of treatment, the study demonstrated statistically significant superiority of B. bifidum SYN-HI-001 compared to placebo with an improvement of the sum score of −101.07 in the verum group compared to −71.24 in the placebo group (P=0.0013). Again, the result was confirmed by the per-protocol analysis (B. bifidum SYN-HI-001: −102.11 vs. placebo: −73.51; P=0.0048) (FIG. 8). Three of the subscales with each score ranging from 0-100 (bowel movement satisfaction, days with pain, and impact on daily life) showed a significant difference in favour of B. bifidum SYN-HI-001, thus strengthening evidence for the positive treatment effect in a symptom-based score. Bowel movement satisfaction was improved by −23.72 in the verum group compared to −16.62 in the placebo group (P=0.0208). Days with pain was improved by −22.71 in the verum group compared to −14.35 in the placebo group (P=0.0080). Impact on daily life was improved by −20.05 in the verum group compared to −14.15 in the placebo group (P=0.0122). The remaining two parameters of the IBS-SSS abdominal pain—severity and severity of bloating—showed a numerically greater reduction in the B. bifidum SYN-HI-001 group.

[0136] 1.6.4.3 Subject's Global Assessment (SGA) of IBS Symptoms, Individual Symptoms and Composite Score 1-4

[0137] The secondary endpoints “SGA”, “abdominal pain”, “distension/bloating” and “urgency” were assessed on a daily basis in the patient diary on a 7-point Likert scale (only abdominal pain was recorded on an 11-point NRS). B. bifidum SYN-HI-001 showed a significant reduction from baseline to the end of treatment of SGA by −0.76 points vs. −0.54 points in the placebo group (P=0.0192), abdominal pain by −1.29 points vs. −0.93 points in the placebo group (P=0.0112) and distension/bloating by −0.69 points vs. −0.50 points in the placebo group (P=0.0456). A composite score 1-4 was calculated for the IBS symptoms (SGA, abdominal pain, distension/bloating and urgency). The patients in the B. bifidum SYN-HI-001 group significantly benefited from the treatment vs. placebo (change from baseline to end of treatment in verum: −1.21 points; placebo: −0.89 points; P=0.0256) (FIG. 9).

[0138] Furthermore, the symptoms “discomfort” and “pain associated with bowel movement” were assessed on a weekly basis in the patient diary on a 7-point Likert scale. Again, B. bifidum SYN-HI-001 showed a significant reduction from baseline to the end of treatment. Discomfort was reduced by −1.35 points vs. −0.92 points in the placebo group (P=0.0015) and pain associated with bowel movement by −0.88 points vs. −0.46 points in the placebo group (P=0.0231) (FIG. 10).

[0139] 1.6.4.4 Health Related Quality of Life (SF-12)

[0140] Evaluation of the SF-12 sum scores showed a significant gain in quality of life in the B. bifidum SYN-HI-001 group. SF-12 sum improved from baseline to the end of treatment by 5.82 in the Bifidobacteria group and by 4.06 in the placebo group (P=0.0382). Mental health sum improved from baseline to the end of treatment by 3.31 in the Bifidobacteria group and by 1.66 in the placebo group (P=0.0309). Physical health sum was numerically larger improved in the verum group (2.51) compared to placebo (0.8965), but the difference did not reach significance. Since IBS patients are rather affected mentally than physically, this result is not surprising (FIG. 11).

[0141] 1.6.4.5 Subgroup Analysis

[0142] Subgroup analysis revealed improvement of the following symptoms which are specific for corresponding IBS types:

[0143] 1. Stool Frequency (IBS-C):

[0144] In IBS-C patients, the mean change from baseline to the end of treatment for frequency of bowel movements showed a significant difference in favor of B. bifidum SYN-HI-001. With B. bifidum SYN-HI-001, a mean increase of 1.66 bowel movements/week was observed in contrast to a decrease of −1.01 bowel movements/week in the placebo group (P=0.0220).

[0145] 2. Loose Stool Form (IBS-D):

[0146] In the subgroup of IBS-D patients, the stool consistency, which was assessed daily in the patient diary via the BSFS (from 1=constipation to 7=diarrhoea), improved from baseline to the end of treatment significantly more in the verum group (−0.68 points) compared to the placebo group (−0.48 points) towards a harder stool consistency (P=0.0939).

[0147] 3. Bowel Movement Satisfaction (IBS-M and IBS-U):

[0148] Both in the IBS-M and IBS-U subgroup, significant differences were found for bowel movement satisfaction in favor of the B. bifidum SYN-HI-001 group. At the end of treatment the difference (verum-placebo) in mean change from baseline was −27.41 [95% CI: −49.26; −5.55] in the IBS-M and −13.71 [95% CI: −24.66; −2.76] in the IBS-U subgroup.

[0149] 1.6.4.6 Adverse Events

[0150] Only 15 adverse events were reported with suspected relationship to the study product, 7 in the B. bifidum SYN-HI-001 and 8 in the placebo group. No significant differences could be detected in the side effects profile of B. bifidum SYN-HI-001 vs. placebo (Table 2).

TABLE-US-00002 TABLE 2 Incidence of Adverse Events with suspected relationship by preferred term (according to MedDRA): B. bifidum SYN-HI-001 Placebo Total N (%) N (%) N (%) P Abdominal distension 1 0.45 1 0.45 2 0.45 10.000 Abdomina pain 2 0.90 1 0.45 3 0.68 0.6233 Abdominal pain upper 0 0.00 1 0.45 1 0.23 10.000 Constipation 0 0.00 1 0.45 1 0.23 10.000 Diarrhoea 0 0.00 2 0.90 2 0.45 0.4989 Gastroenteritis 1 0.45 0 0.00 1 0.23 0.4989 Irritable bowel 1 0.45 0 0.00 1 0.23 0.4989 syndrome aggravated Liver function test 0 0.00 2 0.90 2 0.45 0.4989 Nausea 1 0.45 0 0.00 1 0.23 0.4989 Urinary incontinence 1 0.45 0 0.00 1 0.23 0.4989

[0151] Most adverse events were mild to moderate in severity and resolved after symptomatic treatment. Most of the events were related to gastrointestinal symptoms and can be considered concomitant symptoms of the underlying disease.

[0152] Only two patients reported serious adverse events: acute coronary syndrome and femoral neck fracture, which were both rated as not treatment related and which both occurred in the placebo group. No deaths were reported in this study.

[0153] Medications taken for IBS symptoms were not significantly different between the two treatment groups, as were discontinuations of study treatment. Also laboratory values are not indicative for any safety risk by B. bifidum SYN-HI-001. The evaluation of vital signs and physical examination revealed nothing unusual. Global assessment of tolerability at the end of treatment was rated very good or good in 90.5% of the patients in the B. bifidum SYN-HI-001 and 86.0% in the placebo group (88.3% of the total study population; difference is not significant).

[0154] Overall the study demonstrated good tolerability and did not indicate any safety risk for the use of heat-inactivated B. bifidum SYN-HI-001 in IBS patients.

[0155] The study provided herein shows for the first time that non-viable bacteria of the strain B. bifidum SYN-HI-001 possess in vivo efficacy suitable for the treatment of IBS. Different from probiotic compositions, i.e. compositions based on viable bacteria, these non-viable bacteria of the present invention additionally provide a reduced risk of being associated with adverse side effects, which have previously been reported e.g. in Besselink et al., 2008 for seriously ill subjects. Thus, the non-viable bacteria of the present invention, as well as fragments thereof, provide a promising and safe new tool for the treatment of gastrointestinal disorders.

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