Pharmaceutical composition containing probiotics and prebiotics for preventing or treating drug-resistant infection

Abstract

The present invention provides a pharmaceutical composition comprising probiotics and prebiotics for preventing and / or relieving symptoms associated with infections obtained by extended spectrum beta-lactamase (ESBL)-producing organisms and other drug resistant pathogens. More specifically, the pharmaceutical composition comprises at least one Lactobacillus species as a probiotic and at least one oligosaccharide as a prebiotic. The invention further provides a method of preventing and / or treating infections resulting from extended spectrum beta-lactamase (ESBL)-producing bacteria and other drug resistant pathogens, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a probiotic and a prebiotic.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition comprising probiotics and prebiotics, which is used to prevent mammals from acquiring infections caused by bacteria containing extended-spectrum β-lactamase (ESBL) and other drug-resistant pathogens and / or alleviate symptoms associated with the infection. More specifically, the present invention relates to a composition comprising at least one Lactobacillus species as a probiotic and at least one oligosaccharide as a prebiotic, which is used to prevent mammals from acquiring infections caused by bacteria containing extended-spectrum β-lactamase (ESBL) and other drug-resistant pathogens and / or alleviate symptoms associated with the infection. Background Art

[0002] The background description includes information that may be helpful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, nor is it an admission that any publication specifically or implicitly referenced is prior art.

[0003] An infection is the unwanted colonization of a host organism by a foreign species. Typically, the infecting organism will attempt to use the host's resources to facilitate its own reproduction. As a result, the infecting organism or pathogen may interfere with the normal functioning of the host and may cause more infection-related illnesses of varying severity and, in severe cases, death. Bacterial infections can be treated with appropriate antibiotics. However, choosing the right drug requires determining the type of infection to be treated. Inadvertently using the wrong antibiotic to treat a specific non-viral infection will not treat the infection and may even cause harm. In addition, the widespread use of antibiotics has the potential to lead to the development of antibiotic-resistant infectious species. One means of preventing the rapid spread of resistant pathogens in the human body is to drastically reduce the use of antibiotics. However, before the use of prophylactic antibiotics can be completely stopped, suitable alternatives to antibiotics must be available.

[0004] Antibiotic resistance is a serious global public health problem and is on the rise. Although a few major government health centers have just begun to implement antibiotic stewardship, the misuse of antibiotics continues in both urban and rural areas. Resistant Gram-negative organisms, especially those producing extended-spectrum β-lactamases (ESBLs), have attracted increasing clinical attention in developing countries. ESBLs are mainly found in the Enterobacteriaceae family (Bradford PA: Extended-spectrum β-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin Micro Rev. 2001, 14(4): 933-951), among which Escherichia coli and Klebsiella pneumoniae are the two most common microorganisms carrying ESBL encoding genes: SHV, TEM and CTX-M. The genes carried by these plasmids confer the ability to hydrolyze broad-spectrum cephalosporins and monolactams (Jacoby GA, Munoz-Price LS: The new β-lactamases. N Eng J Med. 2005, 352: 380-391). Some ESBL organisms have also acquired co-resistance to other antibiotics, including co-resistance to fluoroquinolones and aminoglycosides (Isendahl et al, Fecal carriage of ESBL-producing E. coli and K. pneumonia in children in Guinea-Bissau: A hospital-based cross-sectional study. PLoS One. 2010, 7 (12) e51981; Guimaraes et al, Genetic detection of extended-spectrum β-lactamase-containing Escherichia coliisolates and vancomycin-resistant Enterococciin fecal samples of healthy children. Microb Drug Res.2009, 15(3): 211 - 216; Lautenbach et al Epidemiological investigation of fluoroquinolone resistance in infections due to extended spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumonia. Clin Infect Dis. 2001, 33: 1288 - 1294; Ben-Ami et al, A multinational survey of risk factors for infection with extended-spectrum β-lactamase-producing Enterobacteriaceae in nonhospitalized patients. Clin Infect Dis. 2009, 49: 682 - 90), and more recently have also acquired co-resistance to carbapenems (Yong et al Characterization of a new metallo-beta-lactamase gene, bla(NDM-1), and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother. 2009, 53: 5046 - 3054; Pitout et al The latest threat in the war on antimicrobial resistance. Lancet Infect Dis. 2010, 10: 578 - 9; Kamarasamy et al Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study. Lancet Infect Dis.2010,10:597-602; Price et al Clinical epidemiology of the global expansion of Klebsiella pneumoniae carbapenemases. Lancet Infect Dis. 2013,13:185-96), and carbapenemases are the main drugs for treating infections caused by ESBL-producing organisms. .

[0005] Reports from India have shown an increase in ESBL-producing Gram-negative strains in tertiary care hospitals (Lal et al, Occurrence of TEM & SHV gene in extended spectrum β-lactamases (ESBLs) producing Klebsiella sp. isolated from a tertiary care hospital. Indian J Med Res. 2007, 125: 173-178; Jemima et al Multiplex PCR for blaCTX-M & blaSHV in the extended spectrum beta lactamase (ESBL) producing Gram-negative isolates. Indian J Med Res. 2008, 128: 313-317), including outbreaks in neonatal intensive care units (NICUs) (Jain et al, Prevalence of extended-spectrum b-lactamase-producing Gram-negative bacteria in septicaemic neonates in a tertiary care hospital. J Med Microbiol.2003,52:421–425;Cartelle et al,Risk factors forcolonization and infection in a hospital outbreak caused by astrain ofKlebsiella pneumoniae with reduced susceptibility to expanded-spectrumcephalosporins.J Clin Microbiol.2004,42:4242–4249;Lebessi et al,Extended-spectrumβ-lactamase-producing Klebsiella pneumonia in a neonatal intensive care unit in the high-prevalence area of ​​Athens,Greece.J ClinMicrobiol.2002,40:99-804;Royle et al,Outbreak of extended spectrum betalactamase producing Klebsiella pneumoniae in a neonatal unit.Arch Dis ChildFetal Neonatal Ed.1999,80:F64-8;Venezia et al,Molecular epidemiology of anSHV-5extended-spectrum Clin Infect Dis.1995,21:915-23;Jiang et al,Detection of extended-spectrumβ-lactamases in clinical isolatesof Pseudomonas aeruginosa.Antimicrob Ag Chemother.2006,50:2990-2995,Chandelet al,Extended-spectrumβ-lactamase-producing Gram-negative bacteria causingneonatal sepsis in India in rural and urban settings.J Medical Micro.2011,60:500-507) was also developed for the detection of CTX-M-typeβ-lactamase (Pallecchi et al,Detection of CTX-M-typeβ-lactamase). genes in fecal Escherichia coliisolatesfrom healthy children in Bolivia and Peru.Antimicrob Agents Chemother.2004,48(12):4556-4561,Kothari et al,Community acquisition ofβ-lactamase producingEnterobateriaceae in neonatal gut.BMC Microbiology.2013,13:136-141,Rahman et al,High rates of intestinal colonization with extended-spectrum lactamase-producing Enterobacteriaceae among healthy individuals.J Investig Med.2011,59:1284-1286;Luvsansharav et al,Prevalence of risk factors associated with fecal carriage of CTX-Mβ-lactamase-producing Enterobacteriaceae in rural Thai communities.JAntimicrob Chemother.2012,67:1769-1774), the situation is even worse. Although hospital-based studies have shown a high degree of ESBL acquisition and transfer to other patients, there are few data on ESBL stocks in community settings (especially in India). Probiotics are health-promoting bacteria that have been studied as an alternative or supplement to antibiotic therapy (Saavedra JM, Clinical applications of probiotic agents. Am J Clin Nutr. 2001, 73: 1147S-1151S). A recent study in mice showed that probiotics have potential benefits against infections caused by drug-resistant Salmonella enterica serovar Typhimurium DT104 (Asahara et al, Protective effect of Lactobacillus casei strain Shirota against lethal infection with multi-drugresistant Salmonella enterica serovar Typhimurium DT104 in mice. J Appl Microbiol.2011,110:163-73). In humans, during treatment with amoxicillin-clavulanic acid, consumption of cheese containing probiotics can reduce the emergence of resistant enterococci in feces (Bertrand et al, Effect of cheese consumption on emergence of antimicrobial resistance in the intestinalmicroflora induced by a short course of amoxicillin-clavulanic acid. J Appl Microbiol. 2007,102:1052-9). To the best of the knowledge of the inventors of the present disclosure, such activity of probiotics has not been reported in neonates and young infants. .

[0006] Breast milk is often the food of choice for newborns and term infants because of its nutritional content and immunological benefits. However, the nutritional value of raw or conventionally processed donor milk varies and, in most cases, is insufficient to meet the needs of newborns and infants. Recently, certain bacterial strains have attracted a lot of attention because they have been found to display valuable properties if ingested by the human body. In particular, specific strains of the genera Lactobacilli and Bifidobacteria have been found to be able to colonize the intestinal mucosa to reduce the ability of pathogens to adhere to the intestinal epithelium, have immunomodulatory effects and help maintain health. Such bacteria are called probiotics. Prebiotics are sugars that help probiotics grow better in the intestine. However, prebiotics themselves have not been shown to have this effect alone.

[0007] There is a continuing need in the art to develop novel and effective pharmaceutical compositions for neonates, infants, children and adults to treat infections caused by bacteria containing extended-spectrum beta-lactamases (ESBLs) and other resistant pathogens. There is also a need for compositions that provide other benefits, such as compositions that promote intestinal maturation, enhance intestinal health, enhance late protection, promote immune system maturation, help support natural defenses and enhance intestinal comfort while reducing crying time, spasms and / or colic. Summary of the invention

[0008] Purpose of the Invention

[0009] An object of the present invention is to provide a pharmaceutical composition comprising a combination of a prebiotic and a probiotic.

[0010] An object of the present invention is to provide a pharmaceutical composition which can be used to prevent acquisition and / or treat infections caused by bacteria containing extended-spectrum β-lactamases (ESBLs) and other drug-resistant pathogens.

[0011] Another object of the present invention is to provide a pharmaceutical composition that promotes intestinal maturation and enhances intestinal health and subsequent protection in neonates and infants.

[0012] Another object of the present invention is to provide a pharmaceutical composition that promotes maturation of the immune system and helps support natural defenses to enhance intestinal comfort and reduce crying time, spasms and / or colic in newborns and infants.

[0013] Other objects of the present invention will become apparent from the following description of the present invention. SUMMARY OF THE INVENTION

[0015] The present invention relates to a pharmaceutical composition comprising probiotics and prebiotics for preventing infection caused by extended-spectrum β-lactamase (ESBL)-producing organisms and other drug-resistant pathogens in mammals and / or alleviating symptoms associated with the infection. More specifically, the present invention relates to a composition comprising at least one Lactobacillus species as a probiotic and at least one oligosaccharide as a prebiotic for preventing infection caused by extended-spectrum β-lactamase (ESBL)-producing bacteria and other drug-resistant pathogens in mammals and / or alleviating symptoms associated with the infection.

[0016] One aspect of the present disclosure provides a pharmaceutical composition for preventing infection caused by extended-spectrum β-lactamase (ESBL)-producing organisms in a subject or alleviating symptoms associated with the infection, the pharmaceutical composition comprising: a therapeutically effective amount of probiotics; and an effective amount of prebiotics.

[0017] In one embodiment, probiotics include at least one lactobacillus species. In one embodiment, prebiotics include at least one oligosaccharide. In one embodiment, the subject is a human. In one embodiment, the organism producing extended-spectrum beta-lactamase (ESBL) includes gram-negative bacteria producing extended-spectrum beta-lactamase (ESBL). In one embodiment, the at least one lactobacillus species is selected from Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus leichmannii, Lactobacillus plantarum and Lactobacillus cellobiosius. In one embodiment, probiotics include Lactobacillus plantarum. In one embodiment, probiotics include Lactobacillus plantarum strain ATCC202195. In one embodiment, at least one oligosaccharide is selected from the group including the following items: (a) oligofructose (FOS); (b) pectin or pectin polysaccharide; (c) mannan; (d) pentosan, beta-glucan, arabinan or galactan; and (e) mixtures thereof. In one embodiment, at least one oligosaccharide includes oligofructose. In one embodiment, the composition is administered by any one of an oral administration route and a parenteral administration route. In one embodiment, the composition comprises 10-100 billion counts of cells of plant lactobacillus strain ATCC202195 and 100-500 mg of oligofructose. In one embodiment, the composition comprises 10-50 billion counts of cells of plant lactobacillus strain ATCC202195 and 150-350 mg of oligofructose. In one embodiment, the composition further comprises at least one pharmaceutically acceptable excipient.

[0018] Another aspect of the present disclosure relates to a method for preventing a subject in need from acquiring an infection caused by an organism producing extended-spectrum beta-lactamase (ESBL) or alleviating the associated symptoms of the infection, the method comprising administering to the subject a therapeutically effective amount of probiotics and an effective amount of prebiotics. In one embodiment, administration comprises oral or parenteral administration of the probiotics and the prebiotics. In one embodiment, the method comprises administering to the subject a composition comprising a therapeutically effective amount of probiotics and an effective amount of prebiotics. In one embodiment, the composition further comprises at least one pharmaceutically acceptable excipient. In one embodiment, administration comprises simultaneous, continuous or intermittent administration of a therapeutically effective amount of probiotics and an effective amount of prebiotics. In one embodiment, the probiotics include at least one lactobacillus species. In one embodiment, the prebiotics include at least one oligosaccharide. In one embodiment, the at least one lactobacillus species is selected from the group comprising the following items: Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus leichmannii, Lactobacillus plantarum and Lactobacillus cellobiose. In one embodiment, at least one oligosaccharide is selected from the group including the following items: (a) oligofructose (FOS); (b) pectin or pectin polysaccharide; (c) mannan; (d) pentosan, β-glucan, arabinan or galactan; and (e) mixtures thereof. In one embodiment, the therapeutically effective amount of probiotics comprises 10-100 billion counts of cells of Lactobacillus plantarum strain ATCC202195. In one embodiment, an effective amount of prebiotics comprises 100-500 mg oligofructose. In one embodiment, a therapeutically effective amount of probiotics comprises 10-50 billion counts of cells of Lactobacillus plantarum strain ATCC202195. In one embodiment, an effective amount of prebiotics comprises 150-350 mg oligofructose.

[0019] In one embodiment, the prebiotic is selected from the group consisting of: (a) oligosaccharides; (b) fructooligosaccharides ("FOS"), such as soy oligofructose, inulin or banana fiber; (c) pectin or pectin polysaccharides; (d) mannans, such as guar gum, locust bean gum, konjac or xanthan gum; (e) pentosans, β-glucans, arabinans and galactans, such as larch arabinogalactan; and (f) mixtures thereof.

[0020] The compositions of the present disclosure provide a number of health benefits including, but not limited to, promoting intestinal maturation, promoting maturation of the enteric nervous system, enhancing intestinal health, enhancing late protection, promoting immune system maturation, helping to support natural defenses, helping to support growth, enhancing intestinal comfort while reducing crying time, spasms and / or colic, and at least partially meeting the nutritional needs of newborns and infants. DETAILED DESCRIPTION

[0021] The embodiments herein and their various features and advantageous details will be explained more fully with reference to non-limiting embodiments, which will be described in detail in the following description. The description of well-known components and processing techniques is omitted to avoid unnecessarily obscuring the embodiments herein. The examples used herein are merely for the convenience of understanding the manner in which the embodiments herein may be implemented, and further enable those skilled in the art to implement the embodiments herein. Therefore, these examples should not be construed as limiting the scope of the embodiments herein.

[0022] The present invention relates to a pharmaceutical composition comprising probiotics and prebiotics for preventing mammals from acquiring infections caused by extended-spectrum β-lactamase (ESBL)-producing organisms and other drug-resistant pathogens and / or alleviating symptoms associated with the infection. More specifically, the present invention relates to a composition comprising at least one Lactobacillus species as a probiotic and at least one oligosaccharide as a prebiotic for preventing mammals from acquiring infections caused by extended-spectrum β-lactamase (ESBL)-producing bacteria and other drug-resistant pathogens and / or alleviating symptoms associated with the infection.

[0023] One aspect of the present disclosure provides a pharmaceutical composition for preventing a subject from acquiring an infection caused by an extended-spectrum β-lactamase (ESBL)-producing organism or for alleviating symptoms associated with the infection, the composition comprising: a therapeutically effective amount of probiotics; and an effective amount of prebiotics.

[0024] In one embodiment, the probiotics include at least one lactobacillus species. In one embodiment, the prebiotics include at least one oligosaccharide. In one embodiment, the subject is a human. In one embodiment, the organism producing extended-spectrum beta-lactamase (ESBL) includes Gram-negative bacteria producing extended-spectrum beta-lactamase (ESBL). In one embodiment, the at least one lactobacillus species is selected from the group including the following items: Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentation, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus leichmannii, Lactobacillus plantarum and Lactobacillus cellobiose. In one embodiment, the probiotics include Lactobacillus plantarum. In one embodiment, the probiotics include Lactobacillus plantarum strain ATCC202195. In one embodiment, at least one oligosaccharide is selected from the group including the following items: (a) fructooligosaccharides (FOS); (b) pectin or pectin polysaccharides; (c) mannan; (d) pentosan, beta-glucan, arabinan or galactan; and (e) mixtures thereof. In one embodiment, at least one oligosaccharide includes fructooligosaccharides. In one embodiment, the composition is administered by any one of an oral administration route and a parenteral administration route. In one embodiment, the composition comprises a cell of 10-100 billion counts of plant lactobacillus strain ATCC202195 and 100-500mg oligofructose. In one embodiment, the composition comprises a cell of 10-50 billion counts of plant lactobacillus strain ATCC202195 and 150-350mg oligofructose. In one embodiment, the composition further comprises at least one pharmaceutically acceptable excipient.

[0025] Another aspect of the present disclosure relates to a method for preventing an infection caused by an organism producing extended-spectrum beta-lactamase (ESBL) in a subject in need or alleviating symptoms associated with the infection, the method comprising administering a therapeutically effective amount of probiotics and an effective amount of prebiotics to the subject. In one embodiment, administration comprises oral or parenteral administration of the probiotics and the prebiotics. In one embodiment, the method comprises administering a composition comprising a therapeutically effective amount of probiotics and an effective amount of prebiotics to the subject. In one embodiment, the composition further comprises at least one pharmaceutically acceptable excipient. In one embodiment, administration comprises simultaneous, continuous or intermittent administration of a therapeutically effective amount of probiotics and an effective amount of prebiotics. In one embodiment, the probiotics include at least one lactobacillus species. In one embodiment, the prebiotics include at least one oligosaccharide. In one embodiment, the at least one lactobacillus is selected from the group comprising the following items: Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus leichmannii, Lactobacillus plantarum and Lactobacillus cellobiose. In one embodiment, at least one oligosaccharide is selected from the group including the following items: (a) oligofructose (FOS); (b) pectin or pectin polysaccharide; (c) mannan; (d) pentosan, β-glucan, arabinan or galactan; and (e) mixtures thereof. In one embodiment, the therapeutically effective amount of probiotics comprises 10-100 billion counts of cells of Lactobacillus plantarum strain ATCC202195. In one embodiment, an effective amount of prebiotics comprises 100-500 mg oligofructose. In one embodiment, a therapeutically effective amount of probiotics comprises 10-50 billion counts of cells of Lactobacillus plantarum strain ATCC202195. In one embodiment, an effective amount of prebiotics comprises 150-350 mg oligofructose.

[0026] In one embodiment, the prebiotic is selected from the group consisting of: (a) oligosaccharides; (b) fructooligosaccharides ("FOS"), such as soy oligofructose, inulin or banana fiber; (c) pectin or pectin polysaccharides; (d) mannans, such as guar gum, locust bean gum, konjac or xanthan gum, (e) pentosans, β-glucans, arabinans and galactans, such as larch arabinogalactan, and (f) mixtures thereof.

[0027] The compositions of the present disclosure provide a number of health benefits including, but not limited to, promoting intestinal maturation, promoting maturation of the enteric nervous system, enhancing intestinal health, enhancing late protection, promoting immune system maturation, helping to support natural defenses, helping to support growth, enhancing intestinal comfort while reducing crying time, spasms and / or colic, and at least partially meeting the nutritional needs of newborns and infants.

[0028] In some embodiments, the pharmaceutical composition of the present invention can be administered to children aged 1-5 years and adults for the prevention and treatment of infections caused by extended-spectrum β-lactamase (ESBL)-producing bacteria and other drug-resistant pathogens.

[0029] As used herein, the term "probiotic" refers to a microorganism that forms at least a part of a monoculture of transient or endogenous flora, and / or a mixed culture of living or dead microorganisms, spores, parts thereof or metabolites thereof, which exhibits 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.

[0030] Prebiotics are indigestible food ingredients that can stimulate the growth of intestinal bacteria. Pharmaceutical compositions and methods of the present invention include a combination of one or more prebiotics and one or more probiotics. In certain embodiments, these one or more prebiotics include, for example but not limited to, carbohydrates or oligosaccharides and polysaccharides, more preferably oligofructose. The source of oligosaccharides includes fruit, beans and whole grains.

[0031] Oligofructose (FOS) is a long-chain polysaccharide, which is mainly composed of fructose monosaccharides linked together by 1-β-D-fructofuranosyl bonds. After ingestion, oligofructose is only partially hydrolyzed when passing through the mouth, stomach and small intestine. In the large intestine, they become food for some probiotics and are metabolized into short-chain fatty acids, mainly acetic acid, propionic acid, butyric acid and lactic acid. Due to this fermentation, a large number of bacteria are produced. This causes an increase in the number of probiotics, a decrease in intestinal pH, and is believed to inhibit pathogens. The reduction in pH can increase the solubility of calcium and other minerals, and may enhance the absorption of calcium and magnesium. Exemplary oligofructose includes inulin, banana fiber and soy oligofructose, and is present in honey, beer, onions, asparagus, leeks, maple syrup, oats and Jerusalem artichokes.

[0032] The example of suitable probiotic microorganisms can include but is not limited to plant lactobacillus, lactobacillus acidophilus, food lactobacillus (Lactobacillus alimentarius), Lactobacillus casei cheese subspecies (Lactobacillus caseisubsp.casei), Lactobacillus casei Shirota (Lactobacillus casei Shirota), curved lactobacillus (Lactobacillus curvatus), Lactobacillus delbruckiisubsp.lactis (Lactobacillus delbruckiisubsp.lactis), Lactobacil-2Q lus farciminus, Lactobacillus gasseri (Lactobacillus gasseri) and Lactobacillus helveticus (Lactobacillus helveticus).In a preferred embodiment of the present invention, described probiotic is plant lactobacillus.More preferably, described probiotic comprises plant lactobacillus strain ATCC 202195.

[0033] The growth of various Lactobacillus species for forming cell cultures, cell pastes and spore preparations is generally well known in the art. The culture and preparation methods of Lactobacillus plantarum can be readily adapted and / or modified for the growth and preparation of other (lactic acid) producing bacteria disclosed herein.

[0034] Although exemplary of the present invention, Lactobacillus plantarum is used herein merely as a model for various other acid-producing (eg, lactic acid) species of probiotics that are useful in the practice of the present invention and thus should not be considered limiting.

[0035] The term "pharmaceutical composition" as used herein may be interpreted as, but not limited to, a nutritional composition, a nutraceutical composition, a nutritional supplement, or a medicament.

[0036] As used herein, the term "therapeutically effective amount" refers to the amount of active ingredient (ie, probiotic or prebiotic or combination thereof) that elicits a biological or medicinal response in a subject that includes at least partial prevention or treatment of symptoms of the disease being treated or prevented.

[0037] The term "effective amount of a prebiotic" is art-recognized and is used herein to refer to an amount of a prebiotic that promotes the growth and / or maintenance of probiotics administered to a subject such that the prebiotic and probiotics together elicit a desired biological or pharmaceutical response.

[0038] As used herein, the term "WP" refers to a wettable powder, which may be a powder formulation for use as a suspension after being dispersed in water.

[0039] As used herein, the term "SE" refers to Suspo emulsions, which are fluid heterogeneous formulations consisting of solid particles and active ingredients in the form of fine globules in a continuous aqueous phase.

[0040] As used herein, "WG" or "WDG" refers to water dispersible granules, which may be defined herein as a formulation consisting of granules that is used after disintegration and dispersion in a suitable carrier liquid.

[0041] As used herein, "SC" refers to suspension concentrates, which are defined herein as stable suspensions in fluids that are typically diluted with water or other carrier fluid before use.

[0042] As used herein, "WP" refers to wettable powder, which can be a powder formulation for use as a suspension after being dispersed in water or other carrier liquids.

[0043] As used herein, "EC" refers to an emulsifiable concentrate, which is a liquid homogeneous preparation that can be used as an emulsion after dilution in water or other carrier liquids.

[0044] As used herein, "SE" refers to a suspo emulsion, which is a formulation comprising solid and liquid active ingredients dispersed in an aqueous phase or other carrier liquid.

[0045] As used herein, "ZC" refers to stable mixed formulations of capsule suspensions and suspension concentrates in fluids, which are typically prepared for dilution with water or other carrier fluids before use.

[0046] - "Promoting intestinal maturation" refers in particular to (but not limited to) the maturation of the digestive system, including the related maturation of the nervous system and the immune system.

[0047] - "Enhancing intestinal health" or promoting "intestinal comfort" refers in particular to (but not limited to) benefits selected from the following: helping to better balance the intestinal flora, reducing intestinal permeability, reducing spasms, reducing colic, increasing intestinal absorption or selective absorption.

[0048] - "Enhanced protection for later life" refers in particular (but not exclusively) to a reduction in the risk of later infection and / or allergy. Long-term effects of probiotics (e.g. protection against infection or protection against atopic disease).

[0049] - "Promoting the maturation of the immune system" refers in particular (but not limited to) the growth and development of the immune system.

[0050] - "Helps to support natural defenses" refers in particular (but not limited to) to strengthening the immune system, fighting infection, and promoting the maturation of the immune system.

[0051] - "help to support growth" means in particular (but not exclusively) enabling an infant or child to grow as close as possible to the ideal growth curve.

[0052] As used herein, the phrases "parenteral administration" and "parenteral administration" refer to modes of administration other than enteral and topical administration, such as injection, and include, but are not limited to, intravenous, intramuscular, intrapleural, intravascular, intrapericardial, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

[0053] The phrase "pharmaceutically acceptable" is recognized in the art. In certain embodiments, the term includes compositions, polymers and other materials and / or dosage forms that are suitable, within the scope of sound medical judgment, for contact with the tissues of mammals, humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, and commensurate with a reasonable benefit / risk ratio.

[0054] The terms "treating" and "treatment" are art-recognized and include preventing the occurrence of a disease, disorder, or condition in a human or animal that may be a susceptible disease, disorder, and / or condition but has not yet been diagnosed as having the disease, disorder, and / or condition; inhibiting the disease, disorder, or condition, e.g., hindering its progression; and relieving the disease, disorder, or condition, e.g., causing regression of the disease, disorder, and / or condition. Treating a disease or condition includes ameliorating at least one symptom of a particular disease or condition, even if the underlying pathophysiology is not affected, as is well known in the art, and includes administering a composition that reduces the frequency or delays the onset of symptoms of a medical condition in a subject relative to a subject that has not received the composition. The terms "prophylactic or therapeutic" treatment are art-recognized and include administering one or more compositions to a host. If administration is prior to clinical manifestation of an undesirable condition (e.g., a disease or other undesirable state of the host animal), the treatment is prophylactic, i.e., it protects the host from developing the undesirable condition, whereas if administration is after manifestation of the undesirable condition, the treatment is therapeutic (i.e., it is intended to reduce, ameliorate or stabilize the existing undesirable condition or its side effects).

[0055] The phrase "pharmaceutically acceptable excipient" is art-recognized and includes, for example, pharmaceutically acceptable materials, compositions, or vehicles, such as liquid or solid fillers, diluents, solvents, or encapsulating materials, which are involved in delivering or transporting any subject composition from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the subject composition and not injurious to the patient. In certain embodiments, a pharmaceutically acceptable carrier is non-pyrogenic. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and tantalum. soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other nontoxic compatible substances used in pharmaceutical formulations.

[0056] The present invention relates to a pharmaceutical composition for alleviating symptoms associated with infection caused by extended-spectrum beta-lactamase (ESBL)-producing bacteria and other resistant pathogens in newborns and infants (children in the age group of 1 month to 59 months). Preferably, the present invention relates to a pharmaceutical composition comprising a combination of probiotics and prebiotics for treating infections caused by extended-spectrum beta-lactamase (ESBL)-producing bacteria and other resistant pathogens in newborns and infants. In summary, these health benefits include promoting intestinal maturation, promoting intestinal nervous system maturation, enhancing intestinal health, enhancing late protection, promoting immune system maturation, helping to support natural defenses, helping to support growth, enhancing intestinal comfort, reducing crying time, spasms and / or colic, and at least partially meeting the nutritional needs of the infant.

[0057] According to a preferred embodiment of the present invention, the prebiotics include one or more of the following: (a) oligosaccharides, (b) oligofructose ("FOS"), such as soy oligofructose, inulin or banana fiber, (c) pectin or pectin polysaccharide, (d) mannan, such as guar gum, locust bean gum, konjac or xanthan gum, (e) pentosan, β-glucan, arabinan and galactan, such as larch arabinogalactan, and (f) mixtures thereof.

[0058] According to one embodiment, the pharmaceutical composition of the present disclosure can be administered in solid, semisolid or liquid oral dosage forms. In one embodiment of the present disclosure, the pharmaceutical preparation can be in the form of emulsions, solutions, suspensions, syrups, elixirs, tablets, capsules, pills, granules and suppositories. In another embodiment of the present disclosure, the pharmaceutical preparation can be in the form of mixed preparations of water-dispersible granules (WG), suspension concentrates (SC), wettable powders (WP), emulsifiable concentrates (EC), granules, gels, suspended emulsions (SE), capsule suspensions and suspension concentrates (ZC), and is preferably a mixed preparation of water-dispersible granules (WG), suspended emulsions (SE) and capsule suspensions and suspension concentrates (ZC).

[0059] In one embodiment, the pharmaceutical preparation of the present disclosure can be dry. Drying can include spray drying, fluidized bed drying or freeze drying. In a preferred embodiment, the pharmaceutical preparation of the present disclosure is an oral dosage form for sachets of powder or granules, liquids, solutions, suspensions, emulsions or syrups. In another embodiment, the pharmaceutical preparation can include at least one pharmaceutically acceptable excipient selected from the group consisting of the following items: filler, adhesive, diluent, thickener, solvent, coating agent, dispersant, preservative, sweetener, flavoring, defoamer and stabilizer. However, those skilled in the art will appreciate that, without departing from the scope and spirit of the present disclosure, additional pharmaceutically acceptable excipients can be used.

[0060] Pharmaceutically acceptable fillers may be selected from the group consisting of lactose, microcrystalline cellulose, starch, pregelatinized starch, calcium phosphate, calcium sulfate, calcium carbonate, mannitol, sorbitol, xylitol, sucrose, maltose, fructose, dextrose, maltodextrin, etc. Pharmaceutically acceptable binders may be selected from the group consisting of starch, natural sugars, corn sweeteners, natural and synthetic gums, cellulose derivatives, gelatin, povidone, polyethylene glycol, waxes, sodium alginate, alcohols, water, etc. Pharmaceutically acceptable diluents can be selected from the group consisting of calcium carbonate, calcium hydrogen phosphate, calcium phosphate, calcium sulfate, microcrystalline cellulose, microcrystalline silicified cellulose, powdered cellulose, dextrates, dextrose, fructose, lactitol, anhydrous lactose, lactose monohydrate, lactose dihydrate, lactose trihydrate, mannitol sorbitol, starch, pregelatinized starch, sucrose, talc, xylitol, maltodextrin, maltitol, and the like. Pharmaceutically acceptable sweeteners may be selected from the group consisting of alitame, acesulfame potassium, aspartame, D-tryptophan, dextrose, erythritol, fructose, galactose, glycerol, glycyrrhizic acid, glucose, isomalt, xylitol, xylose, lactitol, lactose, levulose, maltitol, maltodextrin, maltol, maltose, mannitol, corn syrup, neohesperidin dihydrochalcone, neotame, saccharin, siclamate, sorbitol, sucralose, sucrose, tagatose, thaumatin, trehalose, etc. Pharmaceutically acceptable flavoring agents may be selected from the group consisting of natural flavoring oils, anethole, acetic acid, ascorbic acid, phosphoric acid, fumaric acid, lactic acid, lemon, linalool, malic acid, menthol, eucalyptol, orange, citric acid, cinnamone, tartaric acid, thymol, vanilla flavor, strawberry, etc. Pharmaceutically acceptable preservatives may be selected from the group consisting of parabens, phenol, chlorocresol, alkyl parabens, benzoic acid and its salts, boric acid and its salts, citric acid and its salts, sorbic acid and its salts, neutral preservatives, mercuric preservatives, quaternary compounds, and the like.

[0061] In another embodiment, pharmaceutical composition of the present invention can also be administered by parenteral route, such as intravenous administration. In one embodiment, pharmaceutical composition of the present invention can include 10-100 billion counted probiotic cells and 100-500mg prebiotics. In one embodiment, said composition comprises 10-50 billion counted plant lactobacillus strain ATCC202195 cells and 150-350mg oligofructose.

[0062] The term "infection caused by extended-spectrum β-lactamase (ESBL)-producing organisms and other resistant pathogens" is recognized in the art and may include urinary tract infections (UTIs), intra-abdominal infections, primary bacteremia, respiratory tract infections, skin and soft tissue infections, but is not limited thereto. In a broad sense, it refers to an infection, ailment or disorder caused by one or more resistant pathogens, such as organisms that produce extended-spectrum β-lactamase (ESBL). In particular, the composition of the present invention can be used to prevent the acquisition and / or treatment of infections caused by resistant pathogens such as organisms that produce extended-spectrum β-lactamase (ESBL), and in particular for infections caused by bacteria that produce extended-spectrum β-lactamase (ESBL), such as Gram-negative bacteria that produce extended-spectrum β-lactamase (ESBL) .

[0063] According to another embodiment, the pharmaceutical composition of the present invention can be administered in a solid or liquid oral dosage form, such as emulsions, solutions, suspensions, syrups, elixirs, tablets, chewable tablets, capsules, pills, granules and suppositories.

[0064] In one embodiment, the composition can be in the form of granules, powder supplements (e.g., supplements that can be mixed with beverages), reconstitutable powders (spray dried, dry mixed, agglomerated), ready-to-eat liquids, bars, and dilutable liquid concentrates, etc. However, without departing from the scope and spirit of the present invention, as known or understood by those skilled in the art, any other solid, liquid or semi-solid composition or formulation may be used to achieve its intended purpose, as described in the present disclosure.

[0065] For parenteral administration, solutions (compositions) can be prepared using, for example, sesame oil or peanut oil, propylene glycol aqueous solutions or sterile aqueous solutions. If desired, this solution can be appropriately buffered, and at first the liquid diluent is made isotonic with enough saline or glucose. These specific aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this respect, the sterile aqueous medium used can be easily obtained by standard techniques well known to those skilled in the art.

[0066] The composition of the present invention is preferably administered to humans and animals in a unit dosage form containing and appropriate amount, such as tablets, capsules, pills, powders, granules, suppositories, sterile parenteral solutions or suspensions, sterile non-parenteral solutions, suspensions and oral solutions or suspensions, etc. For oral administration, solid or liquid unit dosage forms can be prepared.

[0067] In one embodiment, the tablet core comprises one or more hydrophilic polymers. Suitable hydrophilic polymers include, but are not limited to, water-swellable cellulose derivatives, polyalkylene glycols, thermoplastic polyalkylene oxides, acrylic polymers, hydrocolloids, clays, gelling starches, swelling cross-linked polymers and mixtures thereof. Examples of suitable water-swellable cellulose derivatives include, but are not limited to, sodium carboxymethylcellulose, cross-linked hydroxypropyl cellulose, hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), hydroxyisopropyl cellulose, hydroxybutyl cellulose, hydroxyphenyl cellulose, hydroxyethyl cellulose (HEC), hydroxypentyl cellulose, hydroxypropyl ethyl cellulose, hydroxypropyl butyl cellulose and hydroxypropyl ethyl cellulose and mixtures thereof. Examples of suitable polyalkylene glycols include, but are not limited to, polyethylene glycol. Examples of suitable thermoplastic polyalkylene oxides include, but are not limited to, poly (ethylene oxide). Examples of suitable acrylic polymers include, but are not limited to, potassium methacrylate divinylbenzene copolymers, polymethyl methacrylate, high molecular weight cross-linked acrylic acid homopolymers and copolymers, such as those commercially available from Noveon-Chemicals under the trade name CARBOPOLTM. Examples of suitable hydrocolloids include, but are not limited to, alginates, agar, guar gum, locust bean gum, kappa carrageenan, iota carrageenan, tara, gum arabic, tragacanth, pectin, xanthan gum, gellan gum, maltodextrin, galactomannan, pusstulan, laminarin, scleroglucan, gum arabic, inulin, pectin, gelatin, whelan, rhamsan, zooglan, methylan, chitin, cyclodextrin, chitosan, and mixtures thereof. Examples of suitable clays include, but are not limited to, montmorillonites such as bentonite, kaolin, and lithophane; magnesium trisilicate; magnesium aluminum silicate; and mixtures thereof. Examples of suitable gelling starches include, but are not limited to, acid hydrolyzed starches, swelling starches (such as sodium starch glycolate and its derivatives), and mixtures thereof. Examples of suitable swelling cross-linked polymers include, but are not limited to, cross-linked polyvinyl pyrrolidone, cross-linked agar, and cross-linked sodium carboxymethyl cellulose, and mixtures thereof.

[0068] The carrier may comprise one or more suitable excipients for tablet formulations. Examples of suitable excipients include, but are not limited to, fillers, adsorbents, binders, disintegrants, lubricants, glidants, release-modifying excipients, superdisintegrants, antioxidants, or mixtures thereof.

[0069] Suitable binders include, but are not limited to, dry binders such as polyvinylpyrrolidone and hydroxypropylmethylcellulose; wet binders such as water-soluble polymers, including hydrocolloids such as acacia, alginates, agar, guar gum, locust bean, carrageenan, carboxymethylcellulose, tara, gum arabic, tragacanth, pectin, xanthan gum, gellan gum, gelatin, maltodextrin, galactomannan, pusstulan, laminarin, scleroglucan, inulin, whelan, rhamsan, zooglan, methylan, chitin, cyclodextrin, chitosan, polyvinylpyrrolidone, cellulose, sucrose and starch; and mixtures thereof. Suitable disintegrants include, but are not limited to, sodium starch glycolate, cross-linked polyvinylpyrrolidone, cross-linked carboxymethylcellulose, starch, microcrystalline cellulose and mixtures thereof. Suitable lubricants include, but are not limited to, long-chain fatty acids and hydrates or solvates thereof, such as magnesium stearate and stearic acid, talc, glyceride wax and mixtures thereof. Suitable glidants include, but are not limited to, colloidal silicon dioxide. Suitable release modifying excipients include, but are not limited to, insoluble edible materials, pH dependent polymers, and mixtures thereof.

[0070] Suitable insoluble edible materials used as release regulating excipients include, but are not limited to, water-insoluble polymers and low melting point hydrophobic materials, copolymers thereof and mixtures thereof. Examples of suitable water-insoluble polymers include, but are not limited to, ethylcellulose, polyvinyl alcohol, polyvinyl acetate, polycaprolactone, cellulose acetate and derivatives thereof, acrylates, methacrylates, acrylic acid copolymers, copolymers thereof and mixtures thereof. Suitable low melting point hydrophobic materials include, but are not limited to, fats, fatty acid esters, phospholipids, waxes and mixtures thereof. Examples of suitable fats include, but are not limited to, hydrogenated vegetable oils, such as cocoa butter, hydrogenated palm kernel oil, hydrogenated cottonseed oil, hydrogenated sunflower oil and hydrogenated soybean oil, free fatty acids and hydrates or solvates thereof, and mixtures thereof. Examples of suitable fatty acid esters include, but are not limited to, sucrose fatty acid esters, monoglycerides, diglycerides and triglycerides, behenic acid glyceride, palmitic acid glyceride, monostearic acid glyceride, tristearic acid glyceride, trilauric acid glyceride, myristic acid glyceride, GlycoWax-932, lauroyl macrogol-32 glyceride, stearoyl macrogol-32 glyceride and mixtures thereof. Examples of suitable phospholipids include phosphatidylcholine, phosphotidylserene, phosphatidylinositol (phosphotidyl enositol), phosphatidic acid and mixtures thereof. Examples of suitable waxes include, but are not limited to, carnauba wax, spermaceti, beeswax, candelilla wax, shellac wax, microcrystalline wax and paraffin; mixtures containing fats, such as chocolate, and mixtures thereof. Examples of super disintegrants include, but are not limited to, cross-linked sodium carboxymethylcellulose, sodium starch glycolate and cross-linked polyvidone (crospovidone). In one embodiment, the tablet core contains up to about 5wt% of such a super disintegrant.

[0071] Examples of antioxidants include, but are not limited to, tocopherol, ascorbic acid, sodium metabisulfite, butylated hydroxytoluene, butylated hydroxyanisole, edetic acid, hydrates or solvates of edetic acid, and mixtures thereof. Examples of preservatives include, but are not limited to, citric acid, tartaric acid, lactic acid, malic acid, acetic acid, benzoic acid, and sorbic acid, and mixtures thereof.

[0072] In one embodiment, the immediate release coating has an average thickness of at least 50 microns, such as from about 50 microns to about 2500 microns, such as from about 250 microns to about 1000 microns. In an embodiment, the immediate release coating is typically compressed at a density greater than about 0.9 g / cc, as measured by the weight and volume of the particular layer.

[0073] In one embodiment, the immediate release coating comprises a first portion and a second portion, wherein at least one portion comprises a second pharmaceutically active agent. In one embodiment, the portions contact each other at the central axis of the tablet. In one embodiment, the first portion comprises the first pharmaceutically active agent and the second portion comprises the second pharmaceutically active agent.

[0074] In one embodiment, the first portion comprises a first pharmaceutically active agent and the second portion comprises a second pharmaceutically active agent. In one embodiment, one of the portions comprises a third pharmaceutically active agent. In one embodiment, one of the portions comprises a second immediate release portion that is the same pharmaceutically active agent as that contained in the tablet core.

[0075] In one embodiment, the outer coating portion is prepared as a dry blend of materials prior to addition to the coated tablet core. In another embodiment, the outer coating portion comprises dry granules containing the pharmaceutically active agent.

[0076] Preparations with different drug release mechanisms described above can be combined into final dosage forms comprising single or multiple units. Examples of multiple units include multilayer tablets, capsules containing tablets, beads or particles in solid or liquid form. Typical rapid release formulations include, for example, compressed tablets, gels, films, coatings, liquid dosage forms and particles that can be encapsulated in gelatin capsules. Many methods for preparing coatings, coverings or incorporating drugs are known in the art.

[0077] The outer layer of the immediate release dosage, unit dosage form (i.e., tablet), multiple drug-containing beads, granules, or coated core dosage forms contains a therapeutically effective amount of active agent and conventional pharmaceutical excipients. The immediate release dosage unit may be coated or uncoated, and may or may not be mixed with a sustained release dosage unit (e.g., in an encapsulated mixture of immediate release drug-containing granules, granules or beads and sustained release drug-containing granules or beads).

[0078] Sustained-release formulations are usually prepared in the form of diffusion or osmotic systems, for example, as described in the 20th edition of "Remington—The Science and Practice of Pharmacy", Lippincott Williams & Wilkins, Baltimore, Md., 2000. Diffusion systems usually consist of one of two types of devices, i.e., reservoirs and matrices, which are well known and described in the art. Matrix devices are usually prepared by compressing the drug into tablet form with a slowly dissolving polymer carrier.

[0079] The immediate release portion can be incorporated into the sustained release system by applying an accelerated release layer onto a sustained release core using a coating or compression process or by a multiple unit system such as a capsule containing both delayed and immediate release beads.

[0080] Delayed release dosage forms are prepared by coating a solid dosage form with a polymer film that is insoluble in the acidic environment of the stomach but soluble in the neutral or slightly alkaline environment of the small intestine. Delayed release dosage units can be prepared, for example, by coating a drug or drug-containing composition with a selected coating material. The drug-containing composition can be a tablet incorporated into a capsule, a tablet used as the inner core of a "coated core" dosage form, or a variety of drug-containing beads, microparticles or granules for incorporation into tablets or capsules.

[0081] A pulsatile release dosage form is a dosage form that simulates multiple dosing without the need for repeated dosing, typically allowing for at least a two-fold reduction in dosing frequency compared to conventional dosage forms (e.g., solutions or conventional solid dosage forms of immediate-release drugs). A pulsatile release profile is characterized by no release (lag time) or reduced release followed by rapid release of the drug.

[0082] Each dosage form comprises any one or a combination of a therapeutically effective amount of a probiotic and an effective amount of a prebiotic (in the present disclosure, each of "probiotics" and "prebiotics" is independently or synonymously referred to as an "active agent" or "active ingredient"). In one embodiment of a dosage form that simulates a twice-daily dosing profile, about 30 wt% to 70 wt%, preferably 40 wt% to 60 wt% is released in the initial pulse, and correspondingly about 70 wt% to 30 wt%, preferably 60 wt% to 40 wt% is released in the second pulse. For a dosage form that simulates a twice-daily dosing profile, it is preferably released at about 3 hours to less than 14 hours after administration, and more preferably the second pulse is released at about 5 hours to 12 hours.

[0083] Another dosage form comprises a compressed tablet or capsule having an immediate release dosage unit containing a drug, a delayed release dosage unit, and an optional second delayed release dosage unit. In this dosage form, the immediate release dosage unit comprises a plurality of beads, particles, which release the drug substantially immediately after oral administration to provide an initial dose. The delayed release dosage unit comprises a plurality of coated beads or particles, which release the drug about 3 hours to 14 hours after oral administration to provide a second dose.

[0084] Methods for preparing the various pharmaceutical compositions disclosed herein are well within the reach of those skilled in the art. For examples of methods for preparing pharmaceutical compositions, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 19th edition (1995).

[0085] In addition, in certain embodiments, the subject composition of the present application can be lyophilized or adopt another suitable drying technology, such as spray drying. The subject composition can be administered once, or can be divided into multiple smaller doses, administered at different time intervals, which depends in part on the release rate of the composition and the required dosage.

[0086] The method for preparing these preparations or compositions comprises the step of combining the compound with a carrier and optionally one or more auxiliary ingredients. In general, the preparations are prepared by uniformly and intimately combining the compound with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product.

[0087] In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the subject compositions are mixed with one or more pharmaceutically acceptable carriers and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) Disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retarders such as paraffin; (6) absorption promoters such as quaternary ammonium compounds; (7) wetting agents such as acetyl alcohol and glyceryl monostearate; (8) absorbents such as kaolin and bentonite; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical composition may also contain a buffer. Solid ingredients of a similar type may also be used as fillers for soft and hard-filled gelatin capsules using lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0088] Oral liquid dosage forms include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the subject composition, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed oil, corn oil, peanut oil, sunflower oil, soybean oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuran alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof.

[0089] Suspensions, in addition to the active ingredients, may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures thereof.

[0090] In one embodiment, each of the prebiotics and probiotics can be prepared into a separate preparation (e.g., tablets, capsules, powders, etc.), and can be orally administered to a subject in need. As disclosed herein, each prebiotic and probiotic can be administered simultaneously, sequentially or intermittently (e.g., oral ingestion), so as to maintain the beneficial effects of each of the prebiotics and probiotics. It should also be understood that any other active ingredient, herbal medicine, neutral drug or excipient known or understood by those skilled in the relevant art can be co-administered with the composition and / or included in the composition of the present disclosure without departing from the scope and spirit of the present disclosure. The term "co-administration" refers to the administration of more than one active agent so that the duration of the physiological effect of an active substance overlaps with the physiological effect of a second agent. In some embodiments, co-administration includes administering an agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20 or 24 hours of the second agent. Co-administration includes administering two agents simultaneously, approximately simultaneously (e.g., about 1, 5, 10, 15, 20 or 30 minutes apart from each other) or in any order. In some embodiments, co-administration can be achieved by co-formulation, i.e., preparing a single pharmaceutical composition comprising the two substances. In other embodiments, the agents can be formulated separately. In another embodiment, the agents can be linked or conjugated to each other. In one embodiment, the prebiotic and probiotic are formulated into a single formulation (e.g., a tablet, capsule, or powder comprising a desired dose of prebiotics and a desired dose of probiotics). A formulation comprising both a desired dose of prebiotics and a desired dose of probiotics can be a single dose formulation or a multi-dose formulation.

[0091] In one embodiment, a combination of 1 billion counted Lactobacillus plantarum cells and about 150 mg oligofructose, with or without one or more excipients such as maltodextrin, is mixed with 1-2 ml of saline solution or dextrose saline solution (5% DNS) or other suitable carrier liquid, and can be orally given to newborn infants (i.e., given orally to 1-day-old human infants) once a day starting from day 1, and fed daily for 7 days (one week). For the second month, the same treatment regimen can be followed, i.e., a mixture of 1 billion counted Lactobacillus plantarum cells and about 150 mg oligofructose, with or without one or more excipients such as maltodextrin, can be mixed with 1-2 ml of saline solution or dextrose saline solution (5% DNS) or other suitable carrier liquid, and can be orally administered once a day for one week. In the 3rd, 4th and 5th months, a mixture of 2 billion counted Lactobacillus plantarum cells and about 150 mg oligofructose can be mixed with 1-2 ml of saline solution or dextrose saline solution (5% DNS) or other suitable carrier liquid with or without one or more excipients such as maltodextrin or a combination of excipients, and can be taken orally once a day for one week. For the 6th to 12th months, a mixture of 4 billion counted Lactobacillus plantarum cells and about 300 mg oligofructose can be mixed with 3 ml of saline solution or dextrose saline solution (5% DNS) or other suitable carrier liquid with or without one or more excipients such as maltodextrin and can be taken orally once a day for one week. For the 13th to 24th month, a mixture of 5 billion counted Lactobacillus plantarum cells and about 500 mg of oligofructose can be mixed with 5 ml of saline solution or dextrose saline solution (5% DNS) or other suitable carrier liquid, with or without an excipient or combination of excipients such as maltodextrin, and can be taken orally once a day for one week. Alternatively, without departing from the scope and spirit of the present invention, the composition of the present invention can be administered in accordance with any other suitable treatment regimen to obtain the beneficial effects disclosed in the present application. Without departing from the scope and spirit of the present invention, the appropriateness of such a treatment regimen can be determined and / or selected by a doctor based on one or more parameters (including dosage, frequency of administration, route of administration or any changes therein), such as the age of the child / infant, race, complications (e.g., diarrhea, vomiting, etc.), patient compliance, etc., and all such changes in the treatment / treatment regimen will be handled as part of the present disclosure.

[0092] The compositions of the present disclosure provide a number of health benefits including, but not limited to, promoting intestinal maturation, promoting maturation of the enteric nervous system, enhancing intestinal health, enhancing protection against later stages, promoting immune system maturation, helping to support natural defenses, helping to support growth, enhancing intestinal comfort, reducing crying time, spasms and / or colic, and at least partially meeting the nutritional needs of a child.

[0093] Although various embodiments of the present invention have been described above, other and further embodiments of the present invention may be devised without departing from the basic scope of the present invention. The scope of the present invention is determined by the claims which follow. The present invention is not limited to the described embodiments, versions or examples, which are included to enable one of ordinary skill in the art to make and use the present invention when combined with the information and knowledge available to one of ordinary skill in the art.

[0094] Example

[0095] Composition comprising Lactobacillus plantarum ATCC 202195 and oligofructose

[0096] Powder formulations as shown in Table 1 below were prepared, each formulation comprising Lactobacillus plantarum ATCC 202195, oligofructose and maltodextrin.

[0097] Table 1: Powder formulations

[0098]

[0099] Each of the above-mentioned preparations 1 to 3 was packaged in a sachet and stored in a cool, moisture-proof container for further use.

[0100] Administration of Lactobacillus plantarum ATCC 202195 and oligofructose to infants

[0101] A group of infants received a synergistic biologic containing Lactobacillus plantarum ATCC 202195 and oligofructose orally after birth. Neonates were recruited from more than 70 community sites and two tertiary care centers in Odisha, India—Capital Hospital, Bhubaneswar and SCB Medical College, Cuttack. All infants had a gestational age of more than 35 weeks and weighed more than 1800 grams at birth. The group designated as "community infants" were born by normal vaginal delivery and then discharged within 24-48 hours. Infants born in hospitals were delivered by caesarean section and stayed in the hospital for 6-7 days as per the hospital's protocol. After enrollment, neonates were randomly assigned to the synergistic treatment group (oral Lactobacillus plantarum and oligofructose) or placebo group, which was taken orally within the first 24 hours after birth and then daily for 7 consecutive days. Starting from day 1, each infant (treatment group, referred to herein as "symbiotic drug treatment") (i.e., one-day-old human infants) was orally fed with Formulation 1 mixed with 1-2 ml of 5% dextrose saline solution (5% DNS) once a day for 7 days (one week). For the second month, the same treatment method was repeated, i.e., Formulation 1 mixed with 1-2 ml of 5% dextrose saline solution (5% DNS) was orally fed once a day for one week. For the third, fourth and fifth months, each infant was orally fed with Formulation 2 mixed with 1-2 ml of 5% dextrose saline solution (5% DNS) once a day for one week in each of the third, fourth and fifth months. For the sixth to twelfth months, each infant was orally fed with Formulation 3 mixed with 3 ml of 5% dextrose saline solution (5% DNS) once a day for one week in each of the sixth to twelfth months. Stool samples (for both commensal treatment and placebo groups) were randomly selected for examination at three time points (2 / 3 months, 5 / 6 months, and 9 / 12 months).

[0102] Analysis of bacterial isolates

[0103] Stool samples were cultured on primary stool medium to isolate bacterial organisms. No selective medium was used. Gram-negative isolates were identified by rapid test (BioMeiurix). Table 1 below provides the primer sequences used for multiplex PCR.

[0104] Table 1: Primer sequences used for multiplex PCR

[0105]

[0106]

[0107] Molecular characterization of ESBL phenotypes

[0108] DNA Isolation: DNA was also purified directly from infant feces using a commercial kit (Qiagen Inc.).

[0109] Multiplex PCR of ESBL genes: In order to detect ESBL genes simultaneously, published primer pairs for three ESBL genes (SHV, CTX-M and TEM) were used in PCR detection (Jiang et al., Detection of extended-spectrum β-lactamases in clinical isolates of Pseudomonas aeruginosa. Antimicrob Ag Chemother. 2006, 50: 2990-2995; Monstein et al., Multiplex PCR amplification assay for the detection of blaTEM, blaSHV and blaCTX-M genes in Enterobacteriaceae. APMIS. 2007, 115: 1400-08, the contents of both are incorporated herein by reference in their entirety). These specific primer pairs amplified 867, 593 and 445 bp gene targets, respectively. 2 μl of DNA extract was used for each 25 μl PCR reaction. The amplification steps included: initial denaturation at 94°C for 3 minutes; 35 cycles of denaturation at 94°C for 30 seconds, annealing at 58°C for 30 seconds, extension at 72°C for 1 minute, and a final amplification at 72°C for 5 minutes. The collected stool samples were analyzed for ESBL, and the results (number of samples with ESBL) are shown in Tables 2A and 2B below.

[0110] Table 2A: Analysis of ESBLs present in stool samples

[0111]

[0112] Table 2B: Analysis of ESBLs present in stool samples

[0113]

[0114] In conclusion, a significant reduction in the acquisition of ESBL gene-carrying bacteria (particularly ESBL-producing Gram-negative bacteria) was observed in both hospital-born and community-born infants after treatment with symbiotics compared with placebo recipients. These surprising results demonstrate for the first time that oral administration of symbiotics, particularly Lactobacillus plantarum (strain ATCC202195) and a combination of prebiotics in early infancy reduces the acquisition of ESBL gene-carrying bacteria (particularly ESBL-producing Gram-negative bacteria) in both hospital-born and community-born infants.

[0115] Advantages

[0116] The present invention provides a pharmaceutical composition comprising a combination of a prebiotic and a probiotic.

[0117] The present invention provides a pharmaceutical composition for effectively preventing and treating infections caused by organisms producing extended-spectrum beta-lactamases (ESBL) and other drug-resistant pathogens.

[0118] The present invention provides a pharmaceutical composition which promotes intestinal maturation of newborns and infants and enhances intestinal health and later protection.

[0119] The present invention provides a pharmaceutical composition that promotes maturation of the immune system and helps support natural defenses to enhance intestinal comfort while reducing crying time, spasms and / or colic in newborns and infants.

Claims

1. Use of a composition for promoting the maturation of a child or infant's immune system, the composition comprising an effective amount of Lactobacillus plantarum strain ATCC202195.

2. The use of claim 1, wherein the composition further comprises an effective amount of a prebiotic.

3. The use of claim 1 or 2, wherein the composition further comprises an excipient selected from fillers, adsorbents, binders, disintegrants, lubricants, glidants, release-modifying excipients, superdisintegrants, antioxidants or mixtures thereof.

4. The use according to any one of the preceding claims, wherein the effective amount of Lactobacillus plantarum is 1 to 5 billion cell counts.

5. Use according to any of the preceding claims, wherein the effective amount of the prebiotic is 100-500 mg.

6. Use according to any of the preceding claims, wherein the prebiotic comprises at least one oligosaccharide.

7. Use according to any of the preceding claims, wherein the prebiotic comprises one or more oligosaccharides selected from the group consisting of fructooligosaccharides, pectin, pectin polysaccharides, mannans, pentosans, β-glucans, arabinans and galactans or mixtures thereof.

8. Use according to any preceding claim, wherein the prebiotic is fructo-oligosaccharide.

9. Use according to any preceding claim, wherein the child or infant is between 0 and 59 months old.

10. The use according to any one of the preceding claims, wherein the administration is daily for a week, once a month for two months or more.

11. The use according to any one of the preceding claims, wherein the administration is daily for a week, once a month, for three months or longer.

12. The use according to any one of the preceding claims, wherein the administration is daily for a period of one week, once a month, for six months or longer.

13. Use according to any one of the preceding claims, wherein the administration is daily for one week, once a month for twelve months.

14. Use according to any preceding claim, wherein the composition is administered orally.

15. Use according to any preceding claim, wherein the probiotics and prebiotics are administered parenterally.

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