WO2014178007A2

WO2014178007A2 - A pharmaceutical composition comprising a combination of probiotic and prebiotic

Info

Publication number: WO2014178007A2
Application number: PCT/IB2014/061097
Authority: WO
Inventors: Pinaki Panigrahi
Original assignee: Pinaki Panigrahi
Priority date: 2013-05-02
Filing date: 2014-04-30
Publication date: 2014-11-06
Also published as: WO2014178007A3

Abstract

The present invention provides a pharmaceutical composition comprising a probiotic and a prebiotic for alleviating symptoms associated with infections such as neonatal sepsis and pneumonia in neonates and infants. More particularly, the pharmaceutical composition comprises at least one Lactillobacillus specie(s) as a probiotic and at least one oligosaccharide or at least one polysaccharide as a pre-biotic. The present inventon further provides a method of treatment of neonatal sepsisand pneumonia comprising administering an effective amount of the pharmaceutical composition to neonates and infants in need thereof.

Description

TITLE OF THE INVENTION

A PHARMACEUTICAL COMPOSITION COMPRISING A COMBINATION OF

PROBIOTIC AND PREBIOTIC

FIELD OF THE INVENTION

[0001] The present invention relates to a pharmaceutical composition comprising a probiotic and a pre-biotic for alleviating symptoms associated with infections such as neonatal sepsis and pneumonia in neonates and infants. More particularly, the present invention relates to a composition comprising at least one Lactillobacillus specie(s) as a probiotic and at least one oligosaccharide or at least one polysaccharide as a pre-biotic. The present invention also relates to methods of treatment of infections such as neonatal sepsis and pneumonia in neonates and infants.

BACKGROUND OF THE INVENTION

[0002] Human milk is generally a food of choice for neonates and full term infants because of its nutritional composition and immunologic benefits. The source of human milk can be, for example, a donor or the infant's mother. The nutritional value of raw or conventionally-processed donor milk, however, varies and in most instances, is not sufficient to meet the needs of neonates and infants.

[0003] In the recent past, certain strains of bacteria have attracted considerable attention because they have been found to exhibit valuable properties for man if ingested. In particular, specific strains of genera Lactobacilli and Bifidobacteria have been found to be able to colonize the intestinal mucosa, to reduce the capability of pathogenic bacteria to adhere to the intestinal epithelium, to have immunomodulatory effects and to assist in the maintenance of well-being. Such bacteria are called probiotics.

[0004] Prebiotics are sugars that help probiotic bacteria grow better in the intestine. However, prebiotics themselves have not been shown to have such effect alone.

[0005] Extensive studies have been carried out to identify new probiotic strains. For example Vrinda Nair et al, "Probiotics and Prebiotics: Role in Prevention of Nosocomial Sepsis in Preterm Infants, " Volume 2013 (2013), Article ID 874726, 8 pages disclose specific strains of Lactobacilli and Bifidobacteria and their beneficial effects as probiotics for treatment of Sepsis and necrotizing enterocolitis (NEC) in preterm infants and the rational for probiotics and prebiotics therapy with focus on the prevention of nosocomial sepsis in preterm infants. However, Nair fails to disclose a combination of probiotics and prebiotics as a single composition and its use for the treatment of pneumonia and neonatal sepsis in neonates and infants. Sunil Sazawal et al, "Prebiotic and Probiotic Fortified Milk in Prevention of Morbidities among Children: Community-Based, Randomized, Double-Blind, Controlled Trial, " evaluated the efficacy of adding prebiotic oligosaccharide and probiotic Bifidobacterium lactis HN019 into milk, in preventing diarrhea, respiratory infections and severe illnesses, in children aged 1-4 years. Although it discloses the combination of prebiotics and probiotics administered with milk to children to treat some intestinal diseases, it fails to provide a pharmaceutical composition to treat pneumonia and neonatal sepsis in neonates and infants. Neonatal sepsis (blood infection) is a totally different infection in comparison to types of infections and diarrhea in older children (1-5 yr). Respiratory infections in neonates are also different, and many are due to viruses (such as RSV). Even bacterial pathogens in neonates are different from those in children.

[0006] Thus neither Nair nor Sazawal disclose a combination of probiotics and prebiotics as a single composition and its use in treating neonatal sepsis and pneumonia in neonates and infants.

[0007] There is an ongoing need in the art to develop newer and efficacious pharmaceutical compositions for neonates and infants to treat infections such as neonatal sepsis and pneumonia. This new combination should also have benefits such as promoting gut maturation, enhancing gut health, enhancing protection later in life, promoting maturation of the immune system, contributing to support of natural defenses, enhancing gut comfort, reducing crying time, cramps and/or colics.

OBJECTS OF THE INVENTION

[0008] It is an object of the present invention to provide a pharmaceutical composition comprising a combination of a prebiotic and a probiotic.

[0009] It is another object of the present invention to provide a method of treatment of infections such as neonatal sepsis and pneumonia in neonates and infants.

[00010] It is another object of the present invention to provide a pharmaceutical composition comprising a combination of a prebiotic and a probiotic that promotes gut maturation and enhances gut health and protection later in life in neonates and infants. [00011] It is yet another object of the present invention to provide a pharmaceutical composition comprising a combination of a prebiotic and probiotic that promotes maturation of the immune system and contributes to support of natural defences to enhance gut comfort and reduces crying time, cramps and/or colics in neonates and infants.

[00012] Other objects of the present invention will be apparent from the description of the invention herein below.

SUMMARY OF THE INVENTION

[00013] The present invention relates to a pharmaceutical composition comprising a probiotic and a prebiotic for alleviating symptoms associated with infections such as neonatal sepsis and pneumonia in neonates and infants. More particularly, the present invention relates to a pharmaceutical composition comprising at least one Lactillobacillus specie(s) as a probiotic and at least one oligosaccharide or at least one polysaccharide as a pre-biotic.

[00014] The present invention also relates to a method of treatment of infections selected from a group comprising neonatal sepsis and/or pneumonia comprising administering an effective amount of a pharmaceutical composition comprising a probiotic and a pre-biotic to neonates and infants in need thereof.

[00015] In one embodiment, a probiotic comprises one or more of the group comprising

L. acidophilus, L. casei, L. fermentum, L. salivaroes, L. brevis, L. leichmannii, L. plantarum and L. cellobiosius .

[00016] In a preferred embodiment, a probiotic comprises Lactobacillus plantarum. More preferably, a probiotic comprises a. Lactobacillus plantarum strain ATCC 202195.

[00017] In one embodiemt, a prebiotic comprises one or more of the following (a) an oligosaccharide, (b) a fructo-oligosaccharide ("FOS"), such as a soy fructo-oligosaccharide, inulin or banana fiber, (c) a pectin or pectic polysaccharide, (d) a mannan, such as guar gum, locust bean gum, konjac, or xanthan gum, (e) a pentosan, beta-glucan, arabinan and galactan, such as larch arabinogalactan, and (f) mixtures thereof.

[00018] In a preferred embodiment, a prebiotic comprises a fructo-oligosaccharide.

[00019] The pharmaceutical composition of the present invention can be suitable for oral or parenteral administration.

[00020] In an embodiment, the pharmaceutical composition of the present invention can include 1- 10 billion counts of cells of a probiotic and 150-250 mg of prebiotic. [00021] Another embodiment of the present invention is directed to a method of providing health benefits comprising promoting gut maturation, promoting the maturation of the gut nervous system, enhancing gut health, enhancing protection later in life, promoting the maturation of the immune system, contributing to support of natural defenses, contributing to support growth, enhancing gut comfort, reducing crying time, cramps and/or colics, fulfilling at least partially the nutritional requirements of neonates and infants.

BRIEF DESCRITPION OF DRAWINGS

[00022] Figure 1 shows a graph illustrating a total number of bacterial species found in each group.

[00023] Figure 2 shows a graph illustrating a number of gram positive species

[00024] Figure 3 shows a graph illustrating a number of gram negative species

DETAILED DESCRITPION OF INVENTION

[00025] The embodiments herein and the various features and advantageous details thereof are explained more fully with reference to the non-limiting embodiments that are illustrated in the accompanying figures and detailed in the following description. Descriptions of well-known components and processing techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be construed as limiting the scope of the embodiments herein.

[00026] The present invention relates to a pharmaceutical synbiotic composition comprising a probiotic and a pre-biotic for alleviating symptoms associated with infections such as neonatal sepsis and pneumonia in neonates and infants.

[00027] The present invention also relates to a method of treatment of infections selected from a group comprising neonatal sepsis and/or pneumonia comprising administering an effective amount of a pharmaceutical composition comprising a probiotic and a pre-biotic to neonates and infants in need thereof.

[00028] In some embodiments, the pharmaceutical composition of the present invention can also be administered to children of age between 1 to 5 years for treatment of sepsis and pneumonia. [00029] As utilized herein, the term "probiotic" refers to microorganisms that form at least a part of the transient or endogenous flora monoculture, and/or a mixed culture of living or dead microorganisms, spores, fractions thereof, or metabolic products thereof that exhibit a beneficial prophylactic and/or therapeutic effect on the host organism. Probiotics are beneficial bacteria that can be found in various foods, or in the form of dietary supplements.

[00030] As used herein, the term "synbiotic composition" refers to a composition comprising a combination of a probiotic (i.e. at least one Lactillobacillus specie) and prebiotic in a form of synergism.

[00031] Prebiotics are non digestible food ingredients that can stimulate growth of intestinal bacterial growth. The pharmaceutical compositions and methods of the present invention include one or more prebiotics in combination with one or more probiotics. In certain embodiments, these one or more prebiotics include, for example and without limitation, carbohydrates or oligosaccharides and polysachharides, more preferably oligo- fructose. Sources of oligosaccharides can include fruits, legumes, and whole grains.

[00032] Fructo-oligosaccharides (FOS) are long-chain polysaccharides comprised primarily of fructose monosaccharides bonded together by Ι-β-D-fructofuranosyl linkages. Upon ingestion, fructo-oligosaccharides are only partially hydrolyzed as they pass through the mouth, stomach, and small intestine. In the large intestine, they became food for certain probiotics, and are metabolized into short chain fatty acids, mainly acetic, propionic, butyric, and lactic acids. As a consequence of this fermentation, a considerable amount of bacterial mass is produced. This results in increased numbers of probiotic, a lowered intestinal pH, and is believed to inhibit pathogens. A pH decrease will increase solubility of calcium and other minerals and may enhance the absorption of calcium and magnesium. Illustrative fructo- oligosaccharides include inulin, banana fiber, and soy fructo-oligosaccharides, and are found in honey, beer, onion, asparagus, Chinese chive, maple sugar, oats, and Jerusalem artichoke.

[00033] Examples of suitable probiotic micro-organisms can include but not limited to Lactobacillus plantarum Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei subsp. casei, Lactobacillus casei Shirota, Lactobacillus curvatus, Lactobacillus delbruckii subsp. lactis, Lactobacil-2Q lus farciminus, Lactobacillus gasseri, and Lactobacillus helve ticus,.

[00034] In one of the preferred embodiments of the present invention, a pro-biotic is

Lactobacillus plantarum. More preferably, a probiotic comprises a Lactobacillus plantarum strain ATCC 202195. [00035] The growth of various Bacillus species to form cell cultures, cell pastes, and spore preparations is generally well-known within the art. The culture and preparative methods for Lactobacillus plantarum may be readily utilized and/or modified for growth and preparation of the other (lactic) acid-producing bacteria disclosed in the present invention.

[00036] Although exemplary of the present invention, Lactobacillus plantarum is only utilized herein as a model for various other acid-producing (e.g., lactic acid) species of probiotic bacteria which may be useful in the practice of the present invention, and therefore is not to be considered as limiting.

[00037] The term "pharmaceutical composition", as used herein can be construed as but not limited to a nutritional composition, a nutraceutical composition, a nutritional supplement or a pharmaceutical drug.

[00038] The term "neonates" as used herein refer to infants of age between 0-28 days. The term "infants" as used herein refer to infants of age up to 12 months.

[00039] The term "therapeutically effective amount" or "effective amount" as used herein means that amount of active ingredient that elicits the biological or medicinal response in an infant which includes at least partial alleviation of the symptoms of the disease being treated.

[00040] The term "neonatal sepsis" as used herein specifically refers to the presence in a new born baby (neonate) of a bacterial blood stream infection (BSI) (such as meningitis, pneumonia, pyelonephritis or gastroenteritis) in a setting of fever. A number of different bacteria, including Escherichia coli (E.coli), Listeria, and certain strains of streptococcus, may cause neonatal sepsis.

[00041] Pneumonia is more common in early-onset sepsis, whereas meningitis and bacteremia are more common in late-onset sepsis.

[00042] Symptoms of neonatal sepsis related to CNS can include, but are not limited to, lethargy, refusal to suck, meningitis, high pitched cry, excessive crying, convulsions, irritable, hypothermia. Symptoms of neonatal sepsis related to GIT can include, but are not limited to, vomiting, abdominal distension, necrotizing enterocolitis, and blood in stool.

[00043] The present invention relates to a pharmaceutical composition for use in alleviating symptoms associated with neonatal sepsis and pneumonia in neonates and infants. Preferably, the present invention relates to a pharmaceutical composition comprising a combination of a probiotic and a prebiotic to treat neonatal sepsis and pneumonia in neonates and infants. Altogether these health benefits include promoting gut maturation, promoting the maturation of the gut nervous system, enhancing gut health, enhancing protection later in life, promoting the maturation of the immune system, contributing to support of natural defenses, contributing to support growth, enhancing gut comfort, reducing crying time, cramps and/or colics, fulfilling at least partially the nutritional requirements of said infant..

[00044] By "promoting gut maturation" is meant in particular (but not exclusively) maturation of the digestive system, including the related nervous system and immune system.

[00045] By "enhancing gut health" or by promoting "gut comfort" is meant in particular (but not exclusively) benefits selected from contributing to better balance the intestinal flora, reducing cramps, reducing colics, increasing gut absorption or selectivity of absorption.

[00046] By "enhancing protection later in life" is meant in particular (but not exclusively) reducing the risk of infections and/or allergies later in life. The long term effect of probiotics (for example for protection against infections or protection against atopic diseases)

[00047] By "contributing to support of natural defenses" is meant in particular (but not exclusively) enhancing the immune system, fighting infection, enhancing the maturation of the immune system.

[00048] By "contributing to support growth" is meant in particular (but not exclusively) enabling the growth of the infant or children to be as close as possible to the ideal growth curve.

[00049] According to a preferred embodiment of the present invention, a prebiotic comprises one or more of the following (a) an oligosaccharide, (b) a fructo-oligosaccharide ("FOS"), such as a soy fructo-oligosaccharide, inulin or banana fiber, (c) a pectin or pectic polysaccharide, (d) a mannan, such as guar gum, locust bean gum, konjac, or xanthan gum, (e) a pentosan, beta-glucan, arabinan and galactan, such as larch arabinogalactan, and (f) mixtures thereof.

[00050] According to another embodiment, the pharmaceutical composition of the present invention may be administered in solid or liquid oral dosage form such as emulsions, solutions, suspensions, syrups, elixirs tablets, capsules, pills, granules, and suppository. In a yet another embodiment, the pharmaceutical composition of the present invention can also be administered by parenteral route such as intravenous administration.

[00051] The pharmaceutical composition of the present invention can include 1-10 billion count of cells of a probiotic and 150-250 mg of a prebiotic.

[00052] According to another embodiment, the pharmaceutical composition of the present invention may be administered simultaneously, separately or sequentially. [00053] Another embodiment of the present invention is directed to a method of providing health benefits comprising promoting gut maturation, promoting the maturation of the gut nervous system, enhancing gut health, enhancing protection later in life, promoting the maturation of the immune system, contributing to support of natural defences, contributing to support growth, enhancing gut comfort, reducing crying time, cramps and/or colics, fulfilling at least partially the nutritional requirements of infant.

EXAMPLES

[00054] Newborn infants > 35 weeks gestational age and > 1800 grams birth weight who may be likely to be hospitalized for 5-7 days are eligible for study enrollment between 12 and 72 hours of life. Additionally, infants and neonates, had to able to tolerate oral feeds and breastfeeding must have begun by 24 hours of life. Infants with major congenital anomalies, with suspected clinical sepsis, who were on antibiotics or lived >30km away from the hospital were excluded. Informed consent was obtained by at least one parent prior to enrollment. All infants were enrolled at Ispat General Hospital (IGH) in Rourkela, Capital Hospital in Bhubaneswar and SCB Medical College in Cuttack.

[00055] At enrollment, infants were randomly assigned to receive an oral Lactobacillus plantarum preparation or a placebo preparation using a 2: 1 sample allocation (treatment vs. placebo). The study composition was given once a day for 7 days. Active follow-up occurred over a 28 day period either in the hospital or at home. Infants who left the hospital after receiving 4 or more doses of the study composition but before all 7 doses were administered received the remaining doses at home. No additional doses were given at home for those infants who received less than 4 doses in the hospital. Infant stool samples were collected at baseline before study medication was administered (day 1 stool taken on day of life 1-3), and post intervention on day 3 (day of life 2-5), day 7 (day of life 5-9), day of life 14 + 3, day of life 21 + 3, and day of life 28 + 3 days. Stool samples were cultured using standard microbiological and API techniques to identify bacterial strains. Medical events were recorded at each of the six scheduled examinations with weight recorded at baseline and on days 7 and 28. Adverse and serious adverse events were recorded whenever they occurred during the 28 day study period.

[00056] Baseline characteristics, medical events, and adverse events were compared between the two study groups. Mean weight on days 1, 7, and 28 was calculated for infants in the Lactobacillus and placebo groups separately. Percent change from baseline was calculated for each time point as [(weight - day 1 weight) / day 1 weight] x 100 and mean percent change from baseline calculated by day and treatment group.

[00057] Organisms found on infant stool cultures and a number of different bacterial species were examined on days 1, 3, 7, 14, 21, and 28. For 23 infants enrolled after day one of life, the first stool sample taken was entered as the day 3 sample with no baseline sample recorded. However, the date of stool collection was on or before the date study dose 1 was given; therefore, these stool samples were used as baseline samples in the analysis with sample results set to missing for day 3. L. plantarum colonization rates were calculated in each group as the number of stool cultures positive for L. plantarum divided by the number of stool cultures taken on each day. Mean number of species were calculated by day and treatment group. Number of bacterial species was set to zero for negative cultures prior to calculations. Number of species was evaluated for all organisms together and for gram positive and gram negative organisms separately.

[00058] Except as noted, all calculations were completed using SAS version 9.1 software 1. Statistical significance for comparisons between treatment groups on baseline characteristics, medical and adverse events was determined by Fisher's exact test and on percent change in weight from baseline by Wilcoxon rank sum exact tests. For number of bacterial species, tests between day 1 and each of the 3-28 day means within treatment group, as well as between the Lactobacillus and placebo group means at each time point were conducted using longitudinal data analyses. Linear repeated measures models were fit using generalized estimating equation (GEE) methods with variances estimated using the approach described by Zeger and Liang2. Reported p-values are from Wald chi-square tests. Models were fit using SUDAAN Release software.

EXAMPLE 1

Enrollment

[00059] A total of 817 infants were screened and 284 were enrolled in the study. One hundred ninety infants were randomized to receive Lactobacillus plantarum and 94 infants were randomized to receive a placebo preparation for 7 days. Similar numbers were enrolled in each treatment group by site (Tablel).

[00060] One infant randomized to the placebo group (4182123) inadvertently received the pharmaceutical composition. This infant is included in the placebo group for all tables and figures described in following embodiments of the invention. Table 1. Enrollment by Study Site and Treatment Group

EXAMPLE 2

[00061] Study Status: 273 infants completed the 28 day study, 5 infants withdrew and 6 were lost to follow-up (Table2). Two infants from Bhubaneswar (3205101, 3435102) and Cuttack (4103106, 4172104) and one infant from Rourkela (2254113) withdrew. Four infants from Bhubaneswar (3122107, 3228107, 3475109, and 3477103) and two infants from Cuttack (4222119, 4326102) moved from the place of delivery, but followed at day 28 for survival and health status.

Table 2. Study Status at 28 Days by Site

Table 2a. Time from birth date to enrollment date

[00062] Of the infants who completed the study, time from birth to hospital discharge ranged from day 5 of life to day 11. The infant hospitalized for 11 days was in the placebo group. There were 7 infants discharged on day 10, 1 placebo and 6 L. plantarum. The majority in each group were discharged home on day 8.

EXAMPLE 3 Dosing

[00063] The majority of subjects received all 7 study doses, 94% (Table 4). Additionally, 184 (65%) infants received all 7 doses in the hospital before being discharged home (Table 5). The remaining infants received anywhere from 1 to 3 doses at home after discharge (Table 6).

[00064] One infant in the L. plantarum group from Cuttack (4139122) who completed the study received only two doses. The study composition was discontinued due to suspected sepsis which was confirmed with a blood culture (Table 17).

[00065] Of the 5 infants who withdrew: 3 received one dose and the other 2 received two doses.

[00066] Of the 6 infants who moved from enrollment site, 4 received all seven doses; 1 received 6 doses; and 1 received 5 doses.

Table 4. Total Number of Doses Received

5 withdrew from the study (Table 3); 1 stopped due to sepsis

Table 5. Number of Doses Received in the Hospital

Table 6. Number of Doses Received at Home

Total Doses Placebo L. Plantarum Total

0 63 136 199 1 13 23 36

2 18 28 46

3 0 3 3

Total 94 190 284

Demographics

[00067] Baseline demographics are shown by treatment group in Table 7. No statistically significant differences were observed between the groups.

Table 7. Baseline Demographics

Number (percent) or mean ± SD and median (min-max) shown.

Infant Stool Cultures [00068] A baseline sample was obtained for 218 (77%) of the 284 infants enrolled, with most taken on day 2 of life (Table 8).

Table 8. Time from birth date to baseline stool sample

[00069] Stool samples were obtained from the majority of infants in each group on days 3-28. The majority of Day 3 stools were collected on days 3-5 of life (252/276, 91%). Most Day 7 stools were taken on days 6-9 of life (259/275, 94%); the majority of Day 14 stools were taken on days 14-17 of life (252/270, 93%); most Day 21 stools were taken on days 21- 24 of life (252/269, 94%); and most Day 28 stools were taken on days 28-31 of life (245/270, 91%).

[00070] For the majority of infants in both groups, time between receipt of the first study dose and collection of the day 3 stool was 3-4 days (240/254, 94%). Additionally, most infants in both groups, had 7-9 days between receipt of the first study dose and collection of the day 7 stool (209/276, 76%). The remaining 67 infants had 4-6 days between their initial dose and day 7 stool culture.

[00071] In the treated group, 54% to 98% of infants were colonized with L. plantarum during the 3-28 day period (Table 9). Please note that the one placebo infant (4182123) who colonized on days 7 and 14 is the infant randomized to the placebo group who inadvertently received the L. plantarum intervention. This infant received 7 doses of the active intervention.

Table 9: Colonization Rates plus 95% confidence intervals Number of stool samples positive (%) for L. plantarum

Group Baseline Day Day Day Day Day Assignment (Pre-Trt) 3 7 14 21 28

Placebo 0/71 (0%) 0/84 (0%) 1/93 (1%) 1/92 (1%) 0/90 (0%) 0/91

(0%)

L. 0/147 92/169 176/182 175/178 171/179 170/

Plantarum (0%) (54%) (97%) (98%) (96%) 179

(47-62%) (93-99%) (95-99%) (91-98%) (95%

)

(91- 97%)

[00072] The distribution of organisms found on infant stool cultures is shown below by day and treatment group for days 1-28 (Table 10)

Table 10: Com arison of Study Groups by the Distribution of Organisms (Days 1-28)

[00073] The mean number of bacterial species found on infant stool, overall and for gram positive and gram negative organisms separately, is shown by day and treatment group in Table 11. The overall mean number of bacterial species was higher at day 3 and each of the later days compared to day 1 in both the placebo group (p< 0.001 for each day vs. day 1 comparison) and the treated group (p<0.001 for each). The mean number of gram positive species was also higher at day 3 and at the later days compared to day 1 in both the placebo group (p<0.001 for each day vs. day 1 comparison) and the treated group (p<0.001 for each day vs. day 1 comparison) as was the mean number of gram negative species ((p<0.001 for each day vs. day 1 comparison in each group).

[00074] The overall number of bacterial species was higher in the treated group than in the placebo group at each time point after baseline (p<0.01 for each). The mean number of gram positive species was significantly higher in the L. plantarum group than in the placebo group on days 3, 7, 14, 21, and 28 (p<0.001 for each). The mean number of gram negative species was significantly lower in the L. plantarum group at each time point after baseline (p<0.001 for each).

Table 11 : Mean Number of Bacterial Species

Time Species Placebo* L. Plantarum* p-value**

Period (N=94) (N=190)

Dav 1 Gram (+1 0.03 10.021 0.05 (0.021 0.46

Gram (-1 0.62 10.081 0.54 (0.051 0.37

Overall 0.65 10.081 0.59 (0.061 0.55

# infants w/ 71 147

Dav Gram (+1 0.43 (0.061 0.97 (0.061 <0.001

Gram (-1 1.54 10.071 1.25 (0.041 <0.001

Overall 1.96 10.071 2.22 (0.061 0.006

# infants w/ 84 170

I)av 7 Gram (+1 0.96 10.071 1.81 (0.051 <0.001

Gram (-1 1.96 (0.071 1.62 (0.051 <0.001

Overall 2.91 (0.081 3.43 (0.051 <0.001

# infants w/ 93 183

Dav 14 Gram (+1 1.13 (0.081 2.13 (0.051 <0.001

Gram (-1 2.20 (0.081 1.66 (0.051 <0.001

Overall 3.33 (0.091 3.79 (0.061 <0.001

# infants w/ 92 179

Dav 21 Gram (+1 1.28 (0.091 2.29 (0.061 <0.001

Gram (-1 2.23 (0.081 1.64 (0.051 <0.001

Overall 3.51 (0.091 3.94 (0.061 <0.001 # infants w/ 90 180

Dav 28 Gram (+) 1.49 (0.09) 2.30 (0.06) <0.001

Gram (-) 2.26 (0.08) 1.77 (0.05) <0.001

Overall 3.76 (0.10) 4.07 (0.06) 0.009

# infants w/ 91 180

* Mean (SE)

** L. Plantarum vs. placebo by the Wald chi-square test.

* * * Range of p-values depending on variance method and test.

Infant Weight

[00075] Weight was obtained for most infants on days 1, 7, and 28.

[00076] Table 12 below shows mean and median weight by treatment group and study day. Mean and median percent change from baseline is shown for days 7 and 28 by treatment group in Table 13.

[00077] No statistically significant differences were found between the treatment groups in the percent change in weight from baseline.

Table 12. Mean and Median Weight

Table 13. Mean and Median Percent Change from Baseline Weight

Time Placebo L. Plantarum p- Period N Mean ± SE N Mean ± SE value*

Median Median

Day 7 9 2.0 ± 0.6 190 2.6 ± 0.4 0.61

4 2.1 2.1 Day 28 9 29.6 ± 1.6 190 31.5 ± 0.9 0.14

4 28.2 30.7

L. Plantarum vs. placebo by the Wilcoxon rank sum test.

Medical events and Adverse Events

[00078] Table 14 below summarizes medical and adverse events reported during the dosing period, and Table 15 summarizes events reported throughout the 28 day study period. Note that these tables include some events recorded during the study exams (and reported on the HI05 and HI07 forms) which did not meet the strict criteria defined for adverse events, in addition to showing events classified as adverse events (AE) and serious adverse events (SAE).

[00079] Table 16 summarizes the AE and SAEs (reported on the HI 10 and HI10-R) and Table 17 gives details for all AE/SAEs reported. Sepsis was suspected in 8 infants, one in the L. plantarum group and 7 in the placebo group. A blood culture obtained for the infant in the L. plantarum group on day 3 of life was positive for staphylococcus epidermidis, and the infant was treated from day 3 to day 8 of life. Of the 7 placebo infants who underwent a sepsis work-up, 1 had a blood culture positive for E. coli on day 24 of life, 5 had negative blood cultures, and the parents refused blood collection for one infant. All infants, including those with negative cultures, were treated for 2 to >5 days. Note that sepsis was suspected in one additional infant in the L. plantarum group (Rourkela, 2254113) on day 6 of life after study withdrawal on day 5 of life. This infant received 2 study doses. Watery stools and hyperbilirubinemia were reported prior to withdrawal.

[00080] Table 14. Infants with at least one occurrence of the specified event during 7 day dosing p.

L. plantarum vs placebo by Fisher's exact test. Table 15. Infants with at least one occurrence of the specified event during 28 d period

L. plantarum vs placebo by Fisher's exact test.

Table 16. Summar of AE/SAEs Re orted

L. plantarum vs placebo by Fisher's exact test.

Clinical Sepsis is suspected sepsis that was blood culture negative or parent refused to have a culture taken.

* One event is shown for each AE/SAE form reported in the database. Each form is delineated by a row in Table 17.

Table 17. Summary of All AE/SAEs reported

Day SAE/ Blood

Site Infant # Group of Event Additional Information

Life AE Culture Baby was seen to have icterus upto thighs and was given phototherapy. Next day

4 AE Hyperbilirubinemia No icterus was found to have reduced.

Phototherapy stopped and transferred to mother. The next day baby was pink.

Baby was found to have frequent loose stool, so observed. Frequency and water content increased requiring IV fluid therapy. Antibiotics, Ampicillin (7/21-7/22),

R 2254113 L. Plantarum Ceftriaxone (7/22-7/29) and Gentamycin

(7/21-7/22) started with sepsis workup

Rule out sepsis

being done. Gentamycin stopped and

(no sepsis work-up or Yes;

6 SAE Amikacin (7/24-7/29) started two days later.

culture entered since negative

Breast feeding was however continued after withdrawal)

throughout. Six days later baby's stool was seen to have improved considerably, so transferred to mother. Sepsis work up conducted including blood culture. SEPSIS WAS SUSPECTED AFTER CONSENT WITHDRAWN!

Abdominal distension was observed prior to baby being discharged from hospital after completing study doses. The baby was feeding well, no vomiting, no cry; baby was not discharged, but asked to stay over night

C 4347114 L. Plantarum 8 AE Abdominal Distension No

to be evaluated again the next morning. Feeding continued, the baby was seen at night, and the abdominal distension resolved. Baby discharged with advice to follow-up.

Patient was put on Ampicillin &

Gentamycin (10/26-10/31) from day 3 of

Yes; life to day 8. Blood culture was Gram +ve arum Sepsis work-up; fever,

C 4139122 L. Plant 3 SAE positive cocci, sensitive to Vanco, Cloxa, Imipenem.

vomiting

Staph Epi But the baby became clinically well. Also treated with Cloxacillin, day 3 to 8, sensitive (10/26-10/31).

Admitted at 24 days of life with complaints of loose motion (10-12 times semisolid) with occasional rapid respiration examined to

Sepsis work-up; have no abdominal distension or tachypnoea.

Yes;

R 2229110 Placebo 24 SAE re-hospitalization for Investigation revealed haziness of both basal negative

rapid respiration lung fields. Started on Ampicillin (7/8/06- 7/9/06) and discharged the next day with amoxycillin drops orally. Sepsis work up conducted including blood culture.

Mother noticed purulent discharge from umbilicus at 22 days of life and consulted for treatment. There was local umbilical

R 2275102 Placebo 22 AE Umbilical Discharge No sepsis which was treated with local

antiseptic dressing withbetadine lotion. Umbilical discharge subsided after 3 days of antiseptic dressing

The mother complained of watery motions both in frequency and consistency. There was no vomiting, fever or abdominal distension. The baby was feeding normally.

B 3331119 Placebo 23 AE Diarrhea No

The stool examination revealed no inflammatory cells or RBC. The baby was followed for 2 days and the event resolved automatically. Mother complained of swelling around baby's abdomen just after feeding. Abdomen was swollen above the thoracic cage, loops were not visible. Baby was active. The girth

Placebo 7 AE Abdominal Distension No of abdomen at umbilicus was 27cm. The baby was closely followed, distension was absent in afternoon and the following day. The baby was discharged and nothing found

B 3363101 on follow-up.

The mother compalined of more watery motions both in frequency and consistency. There was no vomiting, fever or abdominal distension. The baby was feeding normal.

Placebo 21 AE Diarrhea No

The stool examination revealed no puscell or RBC. The baby was followed for 2 days by Dr. Manas Ranjan Swain. The event resolved automatically.

On day 28 the baby presented with fever and less interest in feeding. Axillary temperature was 38.2. The remaining clinical findings

Sepsis work-up;

Yes; were normal. Baby admitted and given

B 3405114 Placebo 28 SAE re-hospitalization;

negative injection Ampicillin (6/20-6/23) & feeding poorly

Gentamycin (6/20-6/23) as per hospital protocol. Blood culture was sent. Sepsis work up conducted including blood culture.

Admitted with features of fever, abdominal

Sepsis work-up; distention and lack of interest in feeding. Put re-hospitalization; Yes; on antibiotics, IV fluid and other supportive.

B 3479120 Placebo 24 SAE fever, lethargy, poor positive Sepsis work up completed. Sepsis work up feeding, abdominal E.Coli conducted including blood culture.

distension Ampicillin (8/26-9/4) and Gentamycin

(8/26-9/4).

Baby was feeding well, no vomiting, no

Sepsis work-up; lethargy. Antibiotics continued for two days

Yes;

C 4348111 Placebo 5 AE Abdominal Distension Ampicillin & Gentamycin (5/16-5/18).

negative

and Jaundice Blood culture was negative. Jaundice resolved quickly and baby discharged well.

Poor feeding, projectile vomiting, abdominal distention noted 2 days prior to checkup. Abdominal distention was more but activity

Sepsis work-up; was fair. Hospitalization was advised. Has re-hospitalization; poor Yes; not received any medication at home till

C 4356113 Placebo 28 SAE

feeding, vomiting, negative reporting. Blood collected for culture in abdominal distension Bactec. Ampicillin and Gentamycin started (6/22-6/25). Baby also had Seeborrhoea scalp. Sepsis work up conducted including blood culture.

Baby admitted with fever on day 14,

Sepsis work-up; Yes; swelling over right chest; ND done given

R 2119119 Placebo 14 AE

Abscess right chest wall negative Ampicillin given from 10/2/05 to 10/8/05

and Gentamycin on 10/8/05 only

Admitted as a case of suspected clinical sepsis. Antibiotics continued for 5 days,

No; parent

C 4126112 Placebo 6 SAE Sepsis work-up parents refused Blood collection for culture.

refused

Ampicillin (10/2-10/7) & Gentamycin (10/2-10/7)

Had abdominal distention, but no vomiting,

C 4131123 Placebo 4 AE Abdominal Distension No stool (m), feeding well was followed up next day, resolved. Abdominal distention, passed small amount c 4226107 Placebo 3 AE Abdominal Distension No of stool (meconium), no vomiting, feeding well.

Table 18 Additional Information on AE and SAEs

NETID Birth AE/SA Day of Stool Culture Organism 1 Organism 2 Organism Organism 4

Weight E Stool Result 3

Weight Culture

2254113 2770 2610 No Stool prior to withdrawal

4347114 2027 2233 1 Positive (1) Escherichia coli

7 Positive (1) Escherichia coli Klebsiella Bacteroides

pneumoniae sp.

14 Positive (1) Escherichia coli Bacteroides sp. Enterococc Lactobacillus us faecalis plantarum

21 Positive (1) Lactobacillus Escherichia coli Enterococc Enterococcus plantarum us faecalis faecium

28 Positive (1) Lactobacillus Enterococcus Enterococc Bacteroides plantarum faecalis us faecium sp.

4139122 2557 2557 1 Negative (2)

3 Positive (1) Klebsiella Lactobacillus Bacteroides

pneumoniae plantarum sp.

7 Positive (1) Klebsiella Lactobacillus Enterococc Bacteroides pneumoniae plantarum us faecium sp.

14 Positive (1) Escherichia coli Lactobacillus Enterococc

plantarum us faecium

21 Positive (1) Lactobacillus Bacteroides sp. Enterococc Enterococcus plantarum us faecium faecalis

28 Positive (1) Lactobacillus Enterococcus Bacteroides Escherichia plantarum faecium sp. coli

2229110 3810 4200 1 Positive (1) Escherichia coli

7 Positive (1) Escherichia coli Enterobacter

cloacae

14 Positive (1) Escherichia coli Enterobacter Bacteroides

cloacae sp.

21 Positive (1) Escherichia coli Enterobacter Bacteroides Actinomycet cloacae sp. es sp.

28 Positive (1) Escherichia coli Enterobacter Bacteroides Staphylococc cloacae sp. us sp.

2275102 3470 3800 1 Positive (1) Escherichia coli

7 Positive (1) Escherichia coli Enterobacter

cloacae

14 Positive (1) Escherichia coli Enterobacter Bacteroides

cloacae sp.

21 Positive (1) Escherichia coli Enterobacter Bacteroides

cloacae sp.

28 Positive (1) Escherichia coli Enterobacter Bacteroides

cloacae sp. 3331119 3100 1 Positive (1) Escherichia coli

3 Positive (1) Escherichia coli Enterococcus

faecalis

7 Positive (1) Escherichia coli Enterococcus Staphylococ

faecalis cus

epidermidis

14 Positive (1) Escherichia coli Enterococcus Bacteroides Staphylococc faecalis sp. us

epidermidis

21 Positive (1) Escherichia coli Enterococcus Bacteroides Staphylococc faecalis sp. us

epidermidis

28 Positive (1) Escherichia coli Enterococcus Klebsiella Bacteroides faecalis pneumoniae sp.

3363101 2500 2500 1 Positive (1) Escherichia coli

3 Positive (1) Escherichia coli Bacteroides sp.

7 Positive (1) Escherichia coli Klebsiella Staphylococ Enterococcus pneumoniae cus xylosis faecalis

14 Positive (1) Escherichia coli Klebsiella Bacteroides Enterococcus pneumoniae sp. faecalis

21 Positive (1) Actinomyces israelii Staphylococcus Enterococc Escherichia aureus us faecium coli

28 Positive (1) Escherichia coli Klebsiella Actinomyce Staphylococc pneumoniae s israelii us aureus

3405114 2250 3200 1 Positive (1) Escherichia coli

3 Positive (1) Escherichia coli Bacteroides sp. Klebsiella

pneumoniae

7 Positive (1) Escherichia coli Klebsiella

pneumoniae

14 Positive (1) Actinomyces israelii Escherichia coli Klebsiella Staphylococc pneumoniae us xylosis

21 Positive (1) Escherichia coli Klebsiella Actinomyce Staphylococc pneumoniae s israelii us aureus

3479120 2200 2500 1 Positive (1) Escherichia coli

3 Positive (1) Escherichia coli Actinomyces

israelii

7 Positive (1) Escherichia coli Klebsiella Staphylococ

pneumoniae cus xylosis

14 Positive (1) Escherichia coli Klebsiella Staphylococ Enterococcus pneumoniae cus aureus faecium

21 Positive (1) Escherichia coli Klebsiella Bacteroides Actinomyces pneumoniae sp. israelii

28 Positive (1) Escherichia coli Klebsiella Bacteroides Enterococcus pneumoniae sp. faecium

4348111 3534 3500 1 Positive (1) Klebsiella

pneumoniae

3 Positive (1) Klebsiella Bacteroides sp. Escherichia

pneumoniae coli

7 Positive (1) Klebsiella Escherichia coli Bacteroides

pneumoniae sp.

14 Positive (1) Escherichia coli Klebsiella Enterococc Bacteroides pneumoniae us faecium sp.

21 Positive (1) Escherichia coli Klebsiella Bacteroides

pneumoniae sp.

28 Positive (1) Escherichia coli Citrobacter Eubacteriu Enterococcus youngae m limosum faecium

4356113 2847 4036 1 Positive (1) Escherichia coli Klebsiella

pneumoniae

3 Positive (1) Klebsiella Escherichia coli Bacteroides

pneumoniae sp.

7 Positive (1) Eubacterium Bacteroides sp. Escherichia Klebsiella limosum coli pneumoniae

14 Positive (1) Staphylococcus Escherichia coli Klebsiella

epidermidis pneumoniae

21 Positive (1) Escherichia coli Klebsiella Eubacteriu

pneumoniae m limosum

28 Positive (1) Escherichia coli Klebsiella Staphylococ Eubacterium pneumoniae cus aureus limosum

2119119 3130 2900 1 Positive (1) Escherichia coli

3 Positive (1) Escherichia coli

7 Positive (1) Escherichia coli Staphylococcus

sp.

14 Positive (1) Escherichia coli Bacteroides sp. Enterobacte

r cloacae

21 Positive (1) Escherichia coli Bacteroides sp. Enterobacte

r cloacae

28 Positive (1) Escherichia coli Bacteroides sp. Actinomyce

tes sp.

4126112 1961 1930 3 Positive (1) Escherichia coli Enterococcus

faecalis

7 Positive (1) Klebsiella Serratia Clostridia

pneumoniae marcescens sp.

14 Positive (1) Escherichia coli Enterococcus Clostridia

durans sp.

21 Positive (1) Escherichia coli Klebsiella Clostridia Enterococcus pneumoniae sp. durans

28 Positive (1) Escherichia coli Klebsiella Clostridia

pneumoniae sp.

4131123 2602 1 Positive (1) Acinetobacter

baumannii

3 Positive (1) Klebsiella Actinomyces

pneumoniae israelii

7 Positive (1) Actinomyces israelii Enterococcus Klebsiella Bacteroides faecium pneumoniae sp.

14 Positive (1) Bacteroides sp. Escherichia coli Enterococc

us faecium

21 Positive (1) Escherichia coli Enterococcus Bacteroides Klebsiella faecium sp. pneumoniae

28 Positive (1) Klebsiella Enterococcus Bacteroides Staphylococc pneumoniae faecium sp. us

epidermidis

4226107 2730 2710 3 Positive (1) Escherichia coli Enterococcus

faecalis

7 Positive (1) Eubacterium Escherichia coli Bacteroides Enterococcus limosum sp. durans 14 Positive (1) Actinomyces israelii Escherichia coli Bacteroides Klebsiella sp. pneumoniae

21 Positive (1) Escherichia coli Enterococcus Actinomyce Klebsiella durans s israelii pneumoniae

28 Positive (1) Escherichia coli Enterococcus Actinomyce Bacteroides faecium s israelii sp.

[00081] Table 19 illustrates highly significant effect in sepsis, pneumonia, diarrhea, and other infections.

Table 19: Recent Community-based Clinical Trial Data Using the Pharmaceutical composition of the present invention (L. plantarum+FOS).

S.n Control L. Relative Absolute Relativ Odds 95% p value NNT

0 plantarum risk risk e risk Ratio Conf

+ FOS reduction reductio interval

(% Risk n

reduction)

A Neonatal 61 26 0.55738 0.014925 0.4262 0.435 (0.2761, 3.28E- 67

Sepsis 9 0.6884) 04

including

culture

negatives

Culture 30 19 0.36667 0.004828 0.6333 0.630 0.3537, 0.1502 207 negative 797 2 1.1230

systemic

inflammatio

n - late

onset.

Clinical

sepsis

warranting

hospitalizati

on and IV

antibiotics.

Includes

cluture

negative

cases only

(culture

positives are

counted

separately)

Culture 31 7 0.77419 0.010536 0.2258 0.223 0.0982 , 1.09E- 95 positive 4 0.5084 04 sepsis

Gram 15 4 0.73333 0.004828 0.2667 0.265 0.0879 , 0.019 207

Negative 797 4 0.8008

sepsis

Gram 16 3 0.8125 0.005706 0.1875 0.186 0.0542 , 0.0043 175 positive 76 4 0.6407

sepsis

B B. Lower 169 109 0.35503 0.026339 0.645 0.627 0.4893 , 2.48E- 38

Respiratory 1 0.8037 04

Tract

Infections -

X-ray

All 347 170 0.51009 0.0777 0.4899 0.448 0.3698 1.10E- 13 infections 8 , 0.5446 16 combined - including

diarrhea

Other morbidity

Atopy/rash 7 3 0.57143 0.001755 0.4286 0.427 0.1105 , 0.3432 570

926 8 1.6565

Neonatal 8 4 0.5 0.001755 0.5 0.499 0.1501 , 0.3871 570 jaundice 926 1 1.6599

Wheezing 8 7 0.125 0.000438 0.875 0.874 0.3166 1 2278

982 6 , 2.4159

Other minor 86 69 0.19767 0.007462 0.8023 0.796 0.5768 , 0.1909 134 events 687 2 1.0989

reported that

did not

require

medical

intervention

Seven cases in the treatment and eleven cases in the placebo group with lower respiratory infection were not counted under category "c" due to inclusion in the culture positive sepsis category.

EXAMPLE 4

Effectiveness of Lactobacillus plantarum synbiotic composition of the present invention. Enrollment:

[00082] A total of 7092 infants were screened and 4556 were enrolled in the study. 2278 infants were randomized to receive Lactobacillus plantarum and a placebo preparation for 60 days.

Study composition:

[00083] The Lactobacillus plantarum synbiotic composition was prepared by mixing 10-9 organisms of ATCC 202195 strain of Lactobacillus plantarum and 150 mg of Fructo- oligosachharide (FOS) in maltodextrin. The mixture was then dissolved in 2ml solution of dextrose and saline solution. It was then administered orally to infants.

For placebo, only maltodextrin was given.

Study Status:

[00084] For synbiotic group, 2141 infants completed the study, 19 infants withdrew, 112 were lost to follow-up and 6 were dead (Table 20). For placebo group, 2185 infants completed the study, 13 infants withdrew, 76 were lost to follow-up and 4 were dead (Table 20). Table 20: Study Status at 60 Days

Infant Weight:

[00085] Weight was obtained for most infants on days 1, 7, 28 and 60 of study days.

[00086] Table 21 below shows mean weight by treatment group and study days. No statistically significant differences were found between the treatment groups during the study days.

Table 21: Infant weight

[00087] The relative risk reduction test was carried out between symbiotic and placebo groups. Table 22 below illustrates highly significant effects on infections.

Table 22: Relative Risk Reduction Assessment

Placebo Synbiotic Relative Risk p value NNT

Reduction (%)

Enrolled 2278 2278

Deaths 4 6 -50 0.7536

Death or Sepsis 234 141 40 O.0001 24

(Primary outcome) Neonatal Sepsis including 61 26 56 O.0001 67 culture negs* and

positives

Lower Respiratory Tract 169 109 36 O.0001 38

Infections**

Clinical Sepsis (B+C) 230 135 41 O.0001 24

(As defined in protocol)

*Clinical sepsis warranting hospitalization and > 5 days of IV antibiotics. Includes culture negative cases only (culture positives are counted separately)

** X-ray positive pneumonia and lower respiratory tract infections requiring antibiotics therapy

Seven cases in the treatment and eleven cases in the placebo group with lower respiratory infection were not counted under category "C" due to inclusion in the culture positive sepsis category

[00088] Table 23 below shows effect on culture Systemic inflammation.

Table 23: Relative risk reduction assessment for culture Systemic inflammation

[00089] Table 24 below shows effect on other individual infections.

Table 24: Relative risk reduction assessment for other individual infections

Local infections* 30 15 5 0.0348 152

Abscess, otitis 12 5 58 0.1427 325 media

Omphalitis 15 3 80 0.0074 190

Diarrhoea 60 12 80 <0.0001 47

*Pustules >10, oral thrush, conjunctivitis

[00090] Table 25 below shows effect on other infections.

Table 25: Relative risk reduction assessment for other infections

Includes pneumonia, LRIs, sepsis, local infections, abscess, omphalitis

Thus in view of the above results, the synbiotic composition of the present invention significantly reduced the incidence of clinical sepsis and culture positive sepsis in neonates and infants. The synbiotic composition also significantly reduced LRTI/pneumonia. Apart from blocking bacterial translocation, the composition seems to have other immunomodulatory effects as well. ADVANTAGES

[00091] It is an advantage of the present invention to provide a pharmaceutical composition comprising a combination of a prebiotic and a probiotic.

[00092] It is another advantage of the present invention to provide a method of treatment of infections such as neonatal sepsis and pneumonia in neonates and infants.

[00093] It is another advantage of the present invention to provide a pharmaceutical composition comprising a combination of a prebiotic and a probiotic that promotes gut maturation and enhances gut health and protection later in life in neonates and infants.

[00094] It is an advantage of the present invention to provide a pharmaceutical composition comprising a combination of a prebiotic and probiotic that promotes maturation of the immune system and contributes to support of natural defences to enhance gut comfort and reduces crying time, cramps and/or colics in neonates and infants.

Claims

1. A pharmaceutical composition comprising a probiotic and a prebiotic for alleviating symptoms comprising neonatal sepsis and pneumonia in neonates and infants, wherein the probiotic is selected from at least one Lactillobacillus specie(s) and the prebiotic is selected from at least one oligosaccharide(s) or at least one polysaccharide(s).

2. The pharmaceutical composition of claim 1, wherein said composition comprises 1- 10 billion counts of cells of the probiotic and 150-250 mg of the prebiotic.

3. The pharmaceutical composition of claim 1, wherein the at least one Lactillobacillus specie(s) is selected from the group comprising Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei subsp. casei, Lactobacillus casei Shirota, Lactobacillus curvatus, Lactobacillus delbruckii subsp. lactis, Lactobacil- 2Q lus farciminus, Lactobacillus gasseri, and Lactobacillus helveticus.

4. The pharmaceutical composition of claim 1, wherein the at least one Lactillobacillus specie(s) is Lactobacillus plantarum strain ATCC 202195.

5. The pharmaceutical composition of claim 1, wherein the at least one oligosaccharide(s) or the at least one polysaccharide is selected from the group comprising:

(a) fructo-oligosaccharide;

(a) pectin or pectic polysaccharide;

(b) mannan comprising guar gum, locust bean gum, konjac, or xanthan gum, and

(c) pentosan, beta-glucan, arabinan or galactan comprising larch arabinogalactan.

6. The pharmaceutical composition of claim 1, wherein the at least one oligosaccharide(s) is fructo-oligosaccharide.

7. The pharmaceutical composition of claim 1, wherein said composition is suitable for oral or parenteral administration. need thereof.

9. The method of claim 8, wherein the pharmaceutical composition is administered simultaneously, separately or sequentially.

10. The method of claim 8, wherein the pharmaceutical composition is administered orally or parenterally.