USE OF A COMPOSITION

专利摘要:
composition, uses of a composition and uses of bifidobacterium lactis bl04 The present invention relates to a method of treating or prophylaxis of diseases of the respiratory tract in patients which comprises administering to said patient a composition comprising bifidobacterium lactis bl04 and/or a fermentation product of bifidobacterium lactis bl04 and/or a fermentation product of bifidobacterium lactis bl04 and/or a cell lysate of bifidobacterium lactis bl04.
公开号:BR112013031659B1
申请号:R112013031659-4
申请日:2012-06-08
公开日:2021-08-31
发明作者:David Pyne；Allan Cripps；Nicholas WEST；Sampo Lahtinen
申请人:Dupont Nutrition Biosciences Aps；
IPC主号:

专利说明:

FIELD OF THE INVENTION
[001] The present invention relates to a method and uses of Bifidobacterium lactis BL-04 for the treatment of respiratory tract diseases. BACKGROUND OF THE INVENTION
[002] According to a 2006 report by the Australian Institute of Health and Welfare, upper respiratory tract disease URTI is the second most commonly managed problem in general practice and a major cause of hospital admissions among children (Welfare, 2006). These infections are accompanied by substantial economic expenditures. A quarter of all absences from work were the result of URTI (Leder et al, 2003), while an economic analysis estimated the direct cost of respiratory tract infections at $9 billion in the United States (Dixon, 1985). In many cases, these diseases reduce social interaction, health and well-being (Hashem and Hall, 2003). URTI may have greater clinical importance in population subgroups. Staying free from respiratory disease is important for athletes, as evidence shows that those who remain disease free perform better than their disease-reporting competitors (Pyne et al, 2001). Reducing the occurrence of these infections would appear to have substantial economic and community benefits.
[003] There are several placebo-controlled clinical studies that have examined the efficacy of probiotics alone (de Vrese et al, 2006; Cox et al, 2008) and in combination with other intended preventive agents (Pregliasco et al, 2008) with respect to common diseases of the respiratory tract. These studies indicate that probiotic supplementation reduces the amount, duration, and severity of disease compared to placebo supplements (de Vrese et al, 2005; Winkler et al, 2005; de Vrese et al, 2006). More recently, a study in a pool of athletes (mean age 27.0 ± 11.6 years; mean ± standard deviation, reported training load of about ten hours per week) concluded that regular intake of probiotics appears to be beneficial. for the reduction in the frequency of URTI, which may be related to better maintenance of IgA levels in saliva during a period of training and competition in winter. These studies indicate that probiotic supplementation can be a useful nutritional product to reduce the frequency, duration and severity of common infectious diseases.
[004] The inventors have surprisingly found that the use of Bifidobacterium lactis BL-04 (Barrangou, R. et al; J. Bacteriol. 191: 4144-4151 (2009) provides better probiotic effects in the treatment and prevention of tract diseases respiratory compared to other probiotic bacteria or placebo in a pool of healthy physically active adults. BRIEF DESCRIPTION OF THE INVENTION
[005] The present invention is based on the inventors' surprising discovery that the use of the probiotic bacterium Bifidobacterium lactis BL-04 (available through DuPont Nutrition BioSciences Aps (formerly Danisco A/S) from Langebrogade 1, PO Box 17, DK - 1001 Copenhagen K, Denmark) in the treatment and/or prophylaxis of respiratory tract diseases provides beneficial results. It is particularly surprising that this bacterium has improved therapeutic effect compared to other probiotic bacteria and mixtures of probiotic bacteria.
[006] Bifidobacterium lactis BL-04 is also known as Bifidobacteirum animalis subsp. lactis BL-04; these expressions are used interchangeably herein. Bifidobacterium lactis BL-04 is also known as DGCC2908 and RB4825.
[007] Bifidobacterium lactis BL-04 was originally isolated from a healthy adult fecal sample and is a commercial strain that has been used and deposited in the North American Type Culture Collection (ATCC) safe deposit as strain SD5219 (see Barrangou , R. et al; J. Bacteriol. 191:4144-4151(2009)).
[008] This is a publicly available bloodline.
[009] The inventors have found that administration of Bifidobacterium lactis BL-04 to healthy physically active adults results in improved outcomes with respect to severity, duration and frequency of respiratory tract illnesses. In the prior art, the groups of patients described as benefiting from the administration of probiotic bacteria are generally those with less robust immune systems, that is, children and/or immunocompromised patients. This unexpected finding is surprising, as healthy physically active adults generally have a more robust immune system and thus are only infected by more severe or more virulent infections, which are usually more difficult to fight off.
[010] In one aspect of the present invention, therefore, there is provided a composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 for use in the treatment or prophylaxis of respiratory tract diseases in patients.
[011] In another aspect of the present invention, a method of treatment or prophylaxis of diseases of the respiratory tract is provided which comprises administering an effective amount of a composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a Bifidobacterium lactis BL-04 cell lysate to patients.
[012] In a further aspect of the present invention, the use of Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 in the manufacture of medicines is provided. for the treatment of diseases of the respiratory tract.
[013] In another aspect of the present invention, there is provided a composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 for use in modification of the absorption of medications (such as cold and/or flu medications) in patients.
[014] In another aspect of the present invention, a method of modifying the absorption of medications (such as cold and/or flu medications) in patients is provided, which comprises administering an effective amount of a composition comprising Bifidobacterium lactis BL -04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 to patients.
[015] In a further aspect of the present invention, the use of Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 in the manufacture of medicines is provided. of modifying the absorption of medications (such as cold and/or flu medications) in patients. DETAILED DESCRIPTION OF THE INVENTION
[016] The detailed aspects of the present invention are described below. Some of the detailed aspects are partially discussed in separate chapters. This is presented for ease of reference and is not limiting in any way. All of the embodiments described above are equally applicable to all aspects of the present invention, unless the context specifically indicates otherwise.
[017] As used herein, "respiratory tract disease" designates diseases of the upper and lower respiratory tracts. The term disease, as used herein, is synonymous with the term disorder.
[018] In one embodiment, respiratory tract disease is an upper respiratory tract disease.
[019] Upper respiratory tract disorders include, for example, tonsillitis, otitis, rhinitis media (inflammation of the nasal mucosa); rhinosinusitis or sinusitis (inflammation of the nostrils and paranasal sinuses, including frontal, ethmoid, maxillary, and sphenoid); nasopharyngitis, rhinopharyngitis, or common cold (inflammation of the nostrils, pharynx, hypopharynx, uvula, and tonsils); pharyngitis (inflammation of the pharynx, hypopharynx, uvula, and tonsils); epiglottitis or supraglottitis (inflammation of the upper larynx and upraglottic area); laryngitis (inflammation of the larynx); laryngotracheitis (inflammation of the larynx, trachea and subglottic area).
[020] In one embodiment, respiratory tract disease is selected from the group consisting of one or more of the following: tonsillitis, otitis, rhinitis media (inflammation of the nasal mucosa); rhinosinusitis or sinusitis (inflammation of the nostrils and paranasal sinuses, including frontal, ethmoid, maxillary, and sphenoid); nasopharyngitis, rhinopharyngitis, or common cold (inflammation of the nostrils, pharynx, hypopharynx, uvula, and tonsils); pharyngitis (inflammation of the pharynx, hypopharynx, uvula, and tonsils); epiglottitis or supraglottitis (inflammation of the upper larynx and upraglottic area); laryngitis (inflammation of the larynx); laryngotracheitis (inflammation of the larynx, trachea and subglottic area); and tracheitis (inflammation of the trachea and subglottic area).
[021] In one embodiment of the present invention, the respiratory tract disease is selected from the group consisting of one or more of the following: sore throat, sneezing, stuffy nose, runny nose and coughing.
[022] One or more of a sore throat, sneezing, stuffy nose, runny nose, or cough is a symptom of upper respiratory tract disease.
[023] In one embodiment, the composition according to the present invention can be used for the treatment or prophylaxis of two or more (including three or more; four or more; or all) among sore throat, sneezing, stuffy nose, runny nose or cough, which are symptoms of upper respiratory disease.
[024] One or more of sore throat, sneezing, stuffy nose, runny nose or cough is a symptom of one or more of the group consisting of: tonsillitis, otitis, rhinitis media (inflammation of the nasal mucosa); rhinosinusitis or sinusitis (inflammation of the nostrils and paranasal sinuses, including frontal, ethmoid, maxillary, and sphenoid); nasopharyngitis, rhinopharyngitis, or common cold (inflammation of the nostrils, pharynx, hypopharynx, uvula, and tonsils); pharyngitis (inflammation of the pharynx, hypopharynx, uvula, and tonsils); epiglottitis or supraglottitis (inflammation of the upper larynx and upraglottic area); laryngitis (inflammation of the larynx); laryngotracheitis (inflammation of the larynx, trachea and subglottic area); and tracheitis (inflammation of the trachea and subglottic area).
[025] In one embodiment, respiratory tract disease is a lower respiratory tract disease.
[026] Lower respiratory tract disorders include, for example, bronchitis, acute bronchitis, pneumonia, and lung abscesses.
[027] In one embodiment of the present invention, the respiratory tract disease is selected from the group consisting of one or more of the following: cough with chest congestion and cough with sneezing.
[028] In one embodiment of the present invention, respiratory tract disease is selected from the group consisting of both of the following: cough with chest congestion and cough with sneezing.
[029] One or more of cough with chest congestion and cough with sneezing is a symptom of lower respiratory tract disease.
[030] One or more of cough with chest congestion and cough with sneezing is a symptom of one or more of the group consisting of bronchitis, acute bronchitis, pneumonia, and lung abscesses.
[031] In one embodiment, the present invention relates to the reduction of one or more of the incidence, duration or severity (preferably the incidence) of respiratory tract diseases, such as upper respiratory tract disease or lower respiratory tract disease .
[032] In another embodiment, the present invention relates to reducing the incidence, duration or severity (preferably the incidence) of symptoms of respiratory tract diseases, such as upper respiratory tract disease or lower respiratory tract disease.
[033] Preferably, the reduction in the incidence (number or frequency), duration or severity of respiratory tract diseases or symptoms of respiratory tract diseases occurs after administration of Bifidobacterium lactis BL-04 compared to the incidence (number or frequency ), duration or severity, respectively, of respiratory tract diseases or symptoms of respiratory tract diseases in comparative patients without administration of Bifidobacterium lactis BL-04.
[034] In one embodiment, the composition is particularly effective in reducing the incidence, duration or severity (preferably the incidence) of respiratory tract diseases in patients who exhibit symptoms for more than seven days. Without wishing to be bound by theory, if symptoms are exhibited for more than seven days this indicates severe respiratory tract illness. In one embodiment, the composition of the present is particularly effective in reducing the incidence, duration or severity (preferably the incidence) of severe respiratory tract diseases.
[035] In one embodiment, the term "incidence" designates the amount and/or frequency of respiratory tract illnesses or their symptoms. In a further embodiment, the term "incidence" designates the amount and/or frequency of respiratory tract illnesses or their symptoms that last for at least three days, at least five days, or at least seven days.
[036] In one embodiment, the term "prophylaxis" refers to diseases of the respiratory tract or its symptoms that last for at least three days, at least five days, or at least seven days. In a further embodiment, "prophylaxis" refers to severe respiratory tract illnesses or their symptoms.
[037] The expression "product of fermentation of Bifidobacterium lactis BL-04", as used herein, indicates a composition that results from the cultivation (such as fermentation) of Bifidobacterium lactis BL-04 in an appropriate medium; or one of its supernatants, fractions or components. In one embodiment, the product of fermentation of Bifidobacterium lactis BL-04 is the integral composition that results from the cultivation (such as fermentation) of Bifidobacterium lactis BL-04 in an appropriate medium. The fermentation product can be dried before use.
[038] The fermentation product of Bifidobacterium lactis BL-04, in one embodiment, may comprise viable Bifidobacterium lactis BL-04. The fermentation product of Bifidobacterium lactis BL-04, in another embodiment, can be a cell-free fermentation product. A cell-free fermentation product can be a fermentation product of Bifidobacterium lactis BL-04 that results from cultivating (such as fermentation) Bifidobacterium lactis BL-04 in appropriate media, which has been modified to remove and/or inactivate cells bacteria to provide a cell-free ferment. In another embodiment, the fermentation product of Bifidobacterium lactis BL-04 can comprise non-viable Bifidobacterium lactis BL-04 which can be whole or lysed.
[039] The term "cell free", as used herein, indicates that the fermentation product (preferably the fermented one) is substantially free of viable bacterial cells, typically containing less than about 105 viable bacterial cells/ml of product of fermentation, less than about 104 viable bacterial cells/ml of fermentation product, less than about 103 viable bacterial cells/ml of fermentation product, less than about 102 viable bacterial cells/ml of fermentation product or less than about 10 viable bacterial cells/ml of fermentation product. Preferably, the fermentation product is substantially cell-free, typically containing less than about 105 cells/ml of fermentation product, less than about 104 cells/ml of fermentation product, less than about 103 cells/ml of product of fermentation, less than about 102 cells/ml of fermentation product or less than about 10 cells/ml of fermentation product.
[040] In one aspect, one or more cells can be separated from the fermentation product (such as a fermented one). Such separation can be achieved by any means known in the art, including by means of centrifugation and/or filtration. The fermentation product can, for example, be filtered (one or more times in a multi-step process) to remove components such as particulate matter, cells and the like. Alternatively or additionally, one or more cells and/or one or more spores can be separated from the fermentation product (such as fermented) by means of centrifugation, in order to produce a supernatant. Depending on the speed and duration of centrifugation, the supernatant can be cell-free (i.e., a cell-free supernatant) or the supernatant can contain cells, which can be filtered or further centrifuged to provide cell-free supernatant.
[041] In some respects, the fermentation product of Bifidobacterium lactis BL-04 can be a crude extract from the culture medium.
[042] In some aspects, the fermentation product of Bifidobacterium lactis BL-04 may comprise a mixture of components present after cultivation (such as at the end) of Bifidobacterium lactis BL-04. Thus, the expression product of fermentation can include, in addition to active ingredients, other components such as particulate matter, solids, substrates not used during cultivation, fragments, media and cell residues.
[043] The expression "cell lysate of Bifidobacterium lactis BL-04", as used herein, indicates the cell fragment and fluid produced by lysis of cell(s) of Bifidobacterium lactis BL-04. Preferably, the Bifidobacterium lactis BL-04 cell(s) is(are) isolated prior to lysis.
[044] Preferably, Bifidobacterium lactis BL-04 cells are lysed by the following method: Bifidobacterium lactis BL-04 is cultured at 37 °C in anaerobic medium in MRS (or other suitable culture medium) supplemented with 0, 05% cysteine; bacterial cells are harvested by means of centrifugation (6000 rpm/5 min); the supernatant is aspirated and the pellet is optionally frozen at -70°C; 1.5 ml of bacterial cell culture pellets to which 150 µl of T10E1 (10 mM Tris-HCl, pH 7.5; 1 mM EDTA) are added and vortexed to resuspend the cell pellet; 1 μl of Lysozyme Ready-Lise® (Epicentre, Vol. 10, No. 3, 2003) is added to each pellet of re-suspended bacteria (1 to 1.5 ml of culture); incubation at 37°C for thirty minutes to one night; 1 μl of Proteinase K (50 μg/μl) is diluted in 150 μl of 2X T&C Lysis Solution (both are provided in the MasterPure DNA Purification Kit or sold separately) for every 1 to 1.5 ml of pelleted culture; 150 μl of the Lysis/Proteinase K solution is added to the sample and thoroughly mixed; incubation at 65 °C to 70 °C for fifteen minutes, rapid vortexing every five minutes; cooling the samples to 37 °C; 1 μl of RNase A (5 μg/μl, supplied in the kit or sold separately) is added to each sample and thoroughly mixed; incubation at 37 °C for thirty minutes; samples are placed on ice.
[045] Alternative lysis methods for Gram positive bacteria can be used to lyse Bifidobacterium lactis BL-04 - they are known to those skilled in the art.
[046] Suitably, the fermentation product of Bifidobacterium lactis BL-04 can be a fermentation product that is present in the isolated supernatant phase of a culture of Bifidobacterium lactis BL-04 grown under the following conditions: 37 °C anaerobicly in MRS (or other appropriate medium) supplemented with 0.05% cysteine.
[047] In one embodiment, the fermentation product can be obtained (preferably is obtained) by culturing the bacteria in a culture medium until the culture OD at À600 reaches at least 0.6, preferably 0.6 at 1.5; optional removal of bacteria by means of centrifugation and/or filtration (such as centrifugation at 25°C, five minutes, 3000 g and/or sterile filtration) to result in a cell-free fermented product comprising active ingredient(s) .
[048] Suitably, the fermentation product can be obtained (preferably is obtained) using a MRS culture medium with 1.0% sugar or without sugar. Suitably, the fermentation product can be obtained (preferably is obtained) by culturing the bacteria at 37°C. Suitably, the fermentation product can be obtained (preferably is obtained) by culturing the bacteria in an anaerobic manner.
[049] The cultivation of Bifidobacterium lactis BL-04 can take place in about 1 to about 72 hours (h), about 5 to about 60 hours, about 10 to about 54 hours or 24 to 48 hours .
[050] In one aspect, cultivation can be conducted until nutrient depletion occurs (preferably complete nutrient).
[051] In one aspect, cultivation is carried out for an effective time to reach the stationary phase of bacterial growth.
[052] The temperature during cultivation can be from about 30 to about 50 °C, about 32 to about 40 °C, from about 34 to about 38 °C, or about 37 °C.
[053] The pH during cultivation can be pH from about 5 to about 9, about 5 to about 6, about 6 to about 7 or about 7 to about 8.
[054] In one aspect, cultivation preferably takes place under aeration.
[055] Continuous and batch cultivation are known to those of ordinary skill in the field. The fermentation product according to the present invention can be prepared using continuous or batch cultivation.
[056] Suitably, the fermentation product can be harvested during or at the end of the cultivation process.
[057] In one aspect, the fermentation product according to the present invention is harvested during or at the end of the exponential phase.
[058] In one aspect, the fermentation product according to the present invention is harvested during or at the end of the stationary phase.
[059] In one aspect of the present invention, the fermentation product can be prepared in a barrel under commercial conditions.
[060] In one aspect, the crop is shaken and/or moved during cultivation (such as during fermentation).
[061] In one aspect, the level of oxygenation is monitored and/or controlled during cultivation.
Suitably, the composition comprising Bifidobacterium lactis BL-04 according to the present invention or the fermentation product of Bifidobacterium lactis BL-04 or cell lysate of Bifidobacterium lactis BL-04 may be in the form of bacterial suspension, before or after freezing, or in the form of concentrates, whether in dry, lyophilized or frozen form. Whichever form is used, the strain can be frozen.
[063] In one embodiment, the composition comprising Bifidobacterium lactis BL-04 according to the present invention does not comprise an additional microorganism; it does not comprise, for example, an additional probiotic bacteria.
[064] In one embodiment, the composition according to the present invention consists of Bifidobacterium lactis BL-04, such as together with excipients, diluents or vehicles.
Suitably, the composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 according to the present invention may contain one or more additions. Additives may suitably be added during drying and/or during lyophilization of the composition.
[066] The composition comprising Bifidobacterium lactis BL-04 used according to the present invention may comprise from 106 to 1012 CFU of bacteria/g of composition and, more specifically, from 108 to 1012 CFU of bacteria/g of composition, preferably 109 to 1012 CFU/g for the lyophilized form.
Suitably, the composition comprising Bifidobacterium lactis BL-04 used in accordance with the present invention may be administered in dosage of about 106 to about 1012 CFU of Bifidobacterium lactis BL-04/dose, preferably about 108 to about of 1012 CFU of Bifidobacterium lactis BL-04/dose. The expression “per dose” indicates that this amount of Bifidobacterium lactis BL-04 is supplied to patients per day or by ingestion, preferably per day. If Bifidobacterium lactis BL-04 is to be administered in food (such as a yoghurt), for example, the yoghurt will preferably contain about 108 to 1012 CFU of Bifidobacterium lactis BL-04. Alternatively, however, this amount of Bifidobacterium lactis BL-04 can be divided into several administrations, each consisting of a smaller amount of microbial load, provided that the final amount of Bifidobacterium lactis BL-04 received by the patient at any specific time (such as per 24 hour period) is from about 106 to about 1012 CFU of Bifidobacterium lactis BL-04, preferably 108 to about 1012 CFU of Bifidobacterium lactis BL-04.
According to the present invention, an effective amount of Bifidobacterium lactis BL-04 can be at least 106 CFU microorganism/dose, preferably about 106 to about 1012 CFU microorganism/dose, preferably about 108 to about 1012 CFU microorganism/dose.
[069] In one embodiment, preferably, the Bifidobacterium lactis BL-04 used in accordance with the present invention can be administered at a dosage of about 106 to about 1012 CFU microorganism/day, preferably about 108 to about 1012 microorganism CFU/dose. Thus, the effective amount in the present embodiment can be from about 106 to about 1012 CFU microorganism/day, preferably about 108 to about 1012 CFU microorganism/day.
[070] In one embodiment, preferably, the Bifidobacterium lactis BL-04 used in accordance with the present invention can be administered at a dosage of about 108 to about 1010 CFU/day, preferably about 1x109 to about 3x109 CFU/day , such as about 2x109 CFU microorganism/day.
[071] CFU stands for "colony forming units".
[072] In one embodiment, the composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 according to the present invention can be used to modify the absorption of medications (such as cold and/or flu medications) in patients.
[073] In one realization, the term "modify" indicates "reduce".
[074] Modification and/or reduction of medication indicates modification or reduction compared to placebo control.
[075] The placebo control does not receive administration of composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 according to the present invention . Placebo control can receive administration of sucrose.
[076] The term "medication", as used herein, preferably indicates medication against colds and/or flu. This may include one or more of the following types of medication: sore throat treatment, phlegm treatment, cough treatments, decongestants, antihistamines, fever reducers, analgesics, cough suppressants (antitussives), antibiotics, or expectorants ( to reduce the thickness of the mucus).
[077] Medication may comprise or be one or more of the following: decongestant, antihistamine, fever reducer, analgesic (such as for headaches and/or other pain), non-steroidal anti-inflammatory drugs (NSAIDs), cough suppressants (or cough suppressants), antibiotics or expectorants (to reduce the thickness of mucus).
[078] The analgesic may include one or more of the following active ingredients: acetaminophen, aspirin, or ibuprofen, for example.
[079] The fever reducer may include one or more of the following active ingredients: aspirin, ibuprofen, magnesium salicylate, and naproxen, for example.
[080] The decongestant may include one or more of the following active ingredients: phenylpropanolamine (PPA) and/or pseudoephedrine and/or phenylephrine, for example.
[081] Cough suppressant (or cough suppressant) may comprise folcodin and/or noscoapine, for example.
[082] The expectorant may include one or more of the following active ingredients: guaifenesin, acetylcysteine, or ambroxol, for example.
[083] The antihistamine may include one or more of the following active ingredients: bromopheniramine; chlorpheniramine, dimenehydrinate, diphenhydramine, doxylamine; loratadine; cetrizine or fexofenadine, for example.
[084] The antibiotic may include one or more of the following active ingredients: azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, penicillin, amoxicillin, rimacillin, or ampicillin.
[085] The expression "medication intake", as used herein, indicates the total amount of medications (cold and/or flu) ingested by the patient and/or the total number of days the patient uses medication (cold and /or flu).
[086] In one embodiment, the composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 according to the present invention can be used to reduce the total amount of cold and/or flu medication taken by the patient and/or reduce the total number of days the patient uses cold and/or flu medication (compared, for example, to a placebo control) .
[087] The composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 according to the present invention may be administered in or as a food product or can be administered as a pharmaceutically acceptable composition.
[088] In one embodiment, the methods or uses in accordance with the present invention result in increased phagocytic activity of granulocytes (neutrophils) in the patient. Preferably, this increase in activity is at least 1%, at least 3%, at least 5%, at least 7%, at least 10%, at least 12%, at least 15%, at least 17%, at least 20%, at least 25%, at least 30%, at least 40% or at least 50%.
[089] In one embodiment, the methods or uses in accordance with the present invention result in increased phagocytic activity of monocytes in the patient. Preferably, this increase in activity is at least 1%, at least 3%, at least 5%, at least 7%, at least 10%, at least 12%, at least 15%, at least 17%, at least 20%, at least 25%, at least 30%, at least 40% or at least 50%.
[090] In one embodiment of the present invention, the composition comprising Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 according to the present invention comprises at least one additional probiotic microorganism, preferably at least one additional probiotic bacteria.
[091] In this specification, the term “probiotic micro-organism” is defined as covering any non-pathogenic micro-organism that, when administered live (as viable) in adequate amounts, confers health benefit on the host. These probiotic strains generally have the ability to survive passage through the upper part of the digestive tract. They are non-pathogenic, non-toxic and exert their beneficial effect on health, on the one hand, through ecological interactions with the flora residing in the digestive tract and, on the other hand, through their ability to influence the immune system in a way positive by means of the “GALT” (gut-associated lymphoid tissue). Depending on the definition of probiotic, these microorganisms, when supplied in sufficient quantity, have the ability to progress alive through the intestine, but do not cross the intestinal barrier and their primary effects are therefore induced in the lumen and/or wall of the gastrointestinal tract. They form part of the resident flora during the administration period. This colonization (or transient colonization) allows the probiotic microorganism to exert beneficial effects, such as the repression of potentially pathogenic microorganisms present in the flora and interactions with the intestinal immune system.
[092] In preferred embodiments, the additional probiotic micro-organism is a bacterium, preferably a probiotic lactic acid bacterium and/or probiotic Bifidobacterium.
[093] In some embodiments, the bacterium may be a bacterium of one or more of the following genera: Lactococcus, Streptococcus, Pediococcus, Enterococcus, Leuconostoc, Carnobacterium, Propionibacterium, Bifidobacterium, Lactobacillus, Brevibacterium and Vagococcus. In a preferred embodiment, the at least one additional probiotic microorganism is selected from the genera Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium and Saccharomyces.
[094] Preferably, the additional microorganism to be used in accordance with the present invention is a microorganism that is generally recognized as safe and that is preferably GRAS approved.
[095] The technicians in the subject will easily know specific species and/or strains of microorganisms of the genera described herein that are used in the food and/or agricultural industries and that are generally considered suitable for human and/or animal consumption.
[096] Preferably, the additional micro-organism used according to the present invention is one that is suitable for human and/or animal consumption.
[097] In one embodiment, preferably, the additional micro-organism is of the Lactobacillus genus, of the Bifidobacterium genus or a mixture of these. Suitably, the microorganism may be a strain of the species L. acidophilus, L. curvatus, L. casei, L. paracasei, L. salivarius, B. lactis, B animalis, B. longum and/or B. bifidum. In one embodiment, the microorganism may preferably be a strain from the species L. acidophilus, L. curvatus, L. salivarius and/or B. lactis.
[098] In one embodiment, preferably, the microorganism is of the genus Streptococcus.
[099] In one embodiment, preferably, the microorganism is of the Enterococcus genus.
[0100] Preferably, the microorganism can be a strain of the species B. lactis, such as B. lactis 420 or B. lactis HN019.
[0101] For some embodiments, the additional microorganism may be a mixture of more than one probiotic microorganism (preferably more than one probiotic bacteria); a mixture of more than one lactic acid bacteria; or a mixture of one or more probiotic micro-organisms (preferably probiotic bacteria) and one or more lactic acid bacteria. Preferably, the mixture may comprise one or more strains of Lactobacillus spp and/or Bifidobacterium spp.
[0102] The additional micro-organism, preferably Lactobacillus spp, such as L. acidophilus, L. salivarius and L. curvatus, for example, for use in accordance with the present invention is preferably a Gram positive strain.
[0103] Conveniently, it may be a catalase negative strain, with a homofermentative metabolism that generates the production of lactic acid.
[0104] The additional micro-organism, preferably Lactobacillus spp such as L. acidophilus, L. salivarius and L. curvatus, for example, for use in accordance with the present invention, can also produce a bacteriocin, such as lactacin, active against other microorganisms.
[0105] Preferably, the additional micro-organism, preferably Lactobacillus spp, such as L. acidophilus, L. salivarius and L. curvatus, for example, for use in accordance with the present invention, has good resistance to pepsin, under conditions of Acid pH, good resistance to pancreatin and/or good tolerance to bile salts.
[0106] In one embodiment, the additional micro-organism, preferably Lactobacillus spp, such as L. acidophilus, for example, according to the present invention may be a micro-organism, preferably Lactobacillus spp, such as L. acidophilus, per example, which can be described as "hydrophobic", that is, one that has a strong affinity for polar or non-polar hydrophobic organic solvents such as n-decane, chloroform, hexadecane or xylene.
[0107] The preferred Lactobacillus acidophilus according to the present invention may be Lactobacillus acidophilus PTA-4797. This strain of Lactobacillus acidophilus was registered by Rhodia Chimie, 26, quai Alphonse Le Gallo, 92512 Boulogne-Billancourt Cedex, France, in accordance with the Budapest Treaty, in the North American Type Cultivation Collection (ATCC), in which it is located. registered with the registration number PTA-4797.
[0108] It will be understood that the composition according to the present invention may be formulated as a medicine, food product or food supplement. In a preferred embodiment, the composition is formulated as a food supplement.
[0109] Conveniently, when the product is food, Bifidobacterium lactis BL-04 and/or a fermentation product of Bifidobacterium lactis BL-04 and/or a cell lysate of Bifidobacterium lactis BL-04 remain effective (the bacterium, for example, remains viable) until the normal “expiry” or “sales period” date during which the food product is offered for sale by the retailer. Preferably, the time of effectiveness should extend beyond these dates, until the end of the normal period of freshness, when the deterioration of the food becomes apparent. Desired time periods and normal shelf life will vary from food to food and those of ordinary skill in the art will recognize that storage lifetimes will vary with food type, food size, storage temperatures, processing conditions, packing material and packing equipment.
[0110] Preferably, the methods and uses according to the present invention result in modulation of the expression of at least one cytokine. Preferably, the cytokine is selected from the group consisting of GM-CSF, IL-1RA, IL-6, IL-8, IL-10, TNF-α and INF-y.
[0111] In one embodiment, preferably, Bifidobacterium lactis BL-04 in the composition and/or the fermentation product according to the present invention are viable.
[0112] The term “viable” indicates that a microorganism (bacteria) is metabolically active or capable of differentiating.
[0113] In a preferred embodiment, the composition comprises viable Bifidobacterium lactis BL04.
[0114] In some embodiments, Bifidobacterium lactis BL04 (such as viable cells) is isolated from the medium in which it was cultured or from the fermentation product prior to formation of the composition comprising Bifidobacterium lactis BL04 according to the present invention.
[0115] Patient: The term "patient", as used herein, indicates an animal. Preferably, the patient is a mammal, including, for example, farm animals (including cattle, horses, pigs, chickens and sheep) and humans. In some aspects of the present invention, the animal is a companion animal (including pets), such as a dog or cat. In some aspects of the present invention, the patient may suitably be human.
[0116] In one embodiment, the patient is a human being.
[0117] In one embodiment, the patient may be female.
[0118] In one embodiment, the patient may be male.
[0119] In one realization, the patient is not a child. The term “child”, as used herein, indicates a human being seven years of age or younger.
[0120] In one embodiment, the patient is a human being 8 years of age or older.
[0121] In one embodiment, the patient is a human being 16 years of age or older.
[0122] In one embodiment, the patient is a human being 18 years of age or older.
[0123] In one embodiment, the patient is not an immunocompromised patient.
[0124] In one embodiment, the patient is a healthy patient.
[0125] In one embodiment, the patient is not exercise-induced immunosuppressed.
[0126] In one achievement, the patient is not an athlete; he is not, for example, an elite athlete; he is not, for example, a long-distance runner.
[0127] In one embodiment, the patient is a healthy and physically active adult. Adult, as used herein, may indicate a human being 18 years of age or older.
[0128] In one embodiment, the patient has a fully developed and uncompromised immune system.
[0129] Advantages: The inventors have surprisingly found that the use of Bifidobacterium lactis BL-04 significantly reduces the incidence, duration and/or severity of respiratory tract diseases (such as URTI and/or lower respiratory tract diseases) or respiratory tract disease symptoms compared to other probiotic bacteria or placebo in healthy physically active adults.
[0130] Surprisingly, this effect was found using a single strain of Bifidobacterium (as isolated or without additional microorganism in the composition).
[0131] An advantage of the present invention is the fact that Bifidobacterium lactis BL-04 significantly reduces the incidence, duration and/or severity of URTI and lower respiratory tract diseases; or significantly reduces the incidence, duration and/or severity of URTI symptoms and lower respiratory tract diseases.
[0132] This is the first time that a strain of Bifidobacterium has been shown to have an effect on URTI and lower respiratory tract diseases or on the symptoms of URTI and lower respiratory tract diseases.
[0133] It is important that patients are healthy and physically active adults. This is in contrast to previous studies that have been carried out in children or individuals with immunosuppression (ie, individuals whose immune system is not fully developed or otherwise impaired). Some studies have focused on elite athletes (such as long-distance runners), again where individuals undergo strenuous training and suffer from exercise-induced immunosuppression.
[0134] In the present case, the inventors surprisingly found beneficial effects in individuals without immunosuppression.
[0135] Furthermore, the present inventors have surprisingly found that Bifidobacterium lactis BL-04 can significantly reduce the incidence, duration and/or severity of respiratory tract diseases (such as lower respiratory tract diseases and/or URTI) or the symptoms of respiratory tract diseases, even when not in a symbiotic composition or a composition with vitamins and/or minerals.
[0136] The present inventors have found that the beneficial effects observed in the present can be obtained using only Bifidobacterium lactis BL-04. This can have many advantages, including simplifying supplement production and/or reducing supplement manufacturer and supplement manufacturer costs.
[0137] An additional advantage is the fact that Bifidobacterium lactis BL-04 can be used without additional probiotic bacteria; this has the advantage of simplifying stability issues with respect to the supplement. An advantage of the present invention is, therefore, the fact that the composition comprising Bifidobacterium lactis BL-04 (such as alone or without the presence of another micro-organism) is easy to stabilize, as the technicians in the subject are only concerned with the stability of a single bacterium instead of more than one bacterium.
[0138] Additionally or alternatively, the use of a single bacterial strain in a composition may have the advantage of reducing the cost of using the composition compared to mixed strain compositions.
[0139] A further significant advantage of the present invention is the fact that the inventors have surprisingly found that the use of a composition (comprising Bifidobacterium lactis BL-04) according to the present invention can be employed to modify or reduce the absorption of medication in patients.
[0140] In this way, the present invention can conveniently generate reduction in the use of medication by the individual. This includes reducing the amount of one or more of the following medications used by the individual: decongestant, antihistamine, fever reducer, analgesic (such as for headaches and/or other pain), non-steroidal anti-inflammatory drugs ( NSAIDs), cough suppressants (or cough suppressants), antibiotics, or expectorants (to reduce mucus thickness).
[0141] Medicine: The term "medicine", as used herein with respect to the present invention, encompasses medicines for human and animal use in human and veterinary medicine. In addition, the term "medicament", as used herein, indicates any substance that provides a therapeutic and/or beneficial effect. The term "drug", as used herein, is not necessarily limited to substances that require Approval for Sale, but may include substances that can be used in cosmetics, nutraceuticals, foods (including, for example, feed and beverages) and natural remedies.
[0142] Treatment: It should be appreciated that all references herein to treatment include curative, palliative, and prophylactic treatment.
[0143] Substantially pure form and/or isolated form: For some aspects, the microorganism and/or fermentation product and/or cell lysate according to the present invention may be in substantially pure form or may be in isolated form.
[0144] The expression "substantially pure form" is used to indicate that the microorganism and/or fermentation product and/or cell lysate according to the present invention is present at a high level. When the microorganism and/or fermentation product and/or cell lysate is in substantially pure form, the microorganism and/or fermentation product and/or cell lysate is desirably the predominant component present in a composition. Preferably, it is present at a level of more than 30%, more than 50%, more than 75%, more than 90% or even more than 95%, wherein said level is determined on a dry weight/dry weight basis with relative to the total composition under consideration.
[0145] At very high levels (such as levels of greater than 90%, greater than 95% or greater than 99%), the microorganism and/or fermentation product and/or cell lysate can be considered to be “isolated ”.
[0146] Biologically active substances according to the present invention (including polypeptides, nucleic acid molecules, carbohydrates identified/identifiable through selection, lipids identified/identifiable through selection, portions identified/identifiable through selection etc.) may be provided in a form that is substantially free of one or more contaminants with which the substance may otherwise be associated. Thus, for example, it can be substantially free of one or more potentially contaminating nucleic acid molecules and/or polypeptides. They can be provided in a form that is substantially free of other cellular components (such as cell membranes, cytoplasm etc.). When a composition is substantially free of a given contaminant, the contaminant will be at a low level (such as less than 10% level, less than 5% or less than 1% based on dry weight/dry weight defined above).
[0147] Combination with other components: Bifidobacterium lactis BL04 and/or its fermentation product and/or its cell lysate for use in the present invention can be used in combination with other components. Thus, the present invention also relates to combinations. Bifidobacterium lactis BL04 and/or its fermentation product and/or its cell lysate may be termed herein "composition according to the present invention".
[0148] The combination according to the present invention comprises the composition according to the present invention and another component that is suitable for animal or human consumption and is capable of providing medical or physiological benefit to the consumer.
[0149] Other components of the combinations according to the present invention include polydextrose, such as Litesse® and/or maltodextrin and/or lactitol. These other components can optionally be added to the composition to assist the drying process and aid the survival of the microorganisms.
[0150] Additional examples of other suitable components include one or more of: thickeners, gelling agents, emulsifiers, binders, crystal modifiers, sweeteners (including artificial sweeteners), rheology modifiers, stabilizers, antioxidants, dyes, enzymes, carriers, vehicles , excipients, diluents, lubricating agents, flavoring agents, coloring material, suspension-forming agents, disintegrants, granulation binders, etc. These other components can be natural. These other components can be prepared using chemical and/or enzymatic methods.
[0151] In one embodiment, the microorganism and/or fermentation product and/or its cell lysate can be encapsulated.
[0152] In a preferred embodiment, the microorganism and/or fermentation product and/or its cell lysate for use in the present invention can be used in combination with one or more lipids.
[0153] The micro-organism and/or fermentation product and/or its cell lysate for use in the present invention can be used, for example, in combination with one or more lipid micelles. The lipid micelle can be a simple lipid micelle or a complex lipid micelle.
[0154] The lipid micelle can be aggregation of oriented molecules of amphiphatic substances.
[0155] Lipid micelles can be an aggregate with colloidal dimensions of oriented molecules of amphiphatic substances existing in equilibrium in solution with the chemical substance from which they are formed. Micelles are usually electrically charged. In aqueous solution, the individual molecules of the micellar aggregate are oriented with their polar groups pointing towards the aqueous medium and their hydrophobic portion directed towards the center of the micelle.
[0156] Lipid molecules can comprise a lipid and/or an oil.
[0157] As used herein, the expression "thickener or gelling agent" designates a product that prevents separation by reducing the speed or preventing the movement of particles, whether they are droplets of immiscible liquids, air or insoluble solids. Thickening occurs when individual hydrated molecules cause an increase in viscosity, reducing the speed of separation. Gel formation occurs when hydrated molecules bind to form a three-dimensional network that captures the particles and immobilizes them.
[0158] The term "stabilizer", as used herein, is defined as an ingredient or combination of ingredients that prevents a product (such as a food product) from changing over time.
[0159] The term "emulsifier", as used herein, designates an ingredient (such as a food product ingredient) that prevents the separation of emulsions. Emulsions are two immiscible substances, one present in the form of droplets, contained within the other. Emulsions can consist of oil in water, where the droplet or dispersed phase is oil and the continuous phase is water; or water-in-oil, where water becomes the dispersed phase and the continuous phase is oil. Foams, which are gas-to-liquid, and suspensions, which are solid-to-liquid, can also be stabilized using emulsifiers. Aeration can occur in a three-phase system, where air is captured by liquid oil and then stabilized by agglomerated fat crystals stabilized with an emulsifier. Emulsifiers have a polar group with affinity for water (hydrophilic) and a non-polar group that is attracted to oil (lipophilic). They are adsorbed at the interfaces of the two substances, providing an interfacial film that acts to stabilize the emulsion. The hydrophilic/lipophilic properties of emulsifiers are affected by the structure of the molecule. These properties are identified by the hydrophilic/lipophilic balance (HLB) value. Low HLB values indicate greater lipophilic tendencies, which are used to stabilize water-in-oil emulsions. High HLB values are attributed to hydrophilic emulsifiers, typically used in oil-in-water emulsions. These values are derived from simple systems. As foods often contain other ingredients that affect emulsification properties, HLB values may not always be a reliable guide for selecting emulsifiers.
[0160] As used herein, the term "binder" designates an ingredient (such as a food ingredient) that binds the product together through physical or chemical reaction. During “gelling”, for example, water is absorbed, providing a binding effect. Binders can, however, absorb other liquids, such as oils, keeping them inside the product. In the context of the present invention, binders would typically be used in solid or low moisture products, for example, for bakery products: pastries, donuts, breads and others.
[0161] The term "crystal modifier", as used herein, designates an ingredient (such as a food ingredient) that affects the crystallization of fat or water. Stabilization of ice crystals is important for two reasons. The first is directly related to product stability from the standpoint of separation. The more freeze/thaw cycles a product encounters, the larger the ice crystals become. These large crystals can break down the structure of the product, whether it is naturally occurring, such as in the case of cell walls, or what is created through “elation”. As water is no longer held in place, the product may exhibit syneresis or oozing after thawing. Second, in the case of a product that is consumed frozen, these large crystals result in an unwanted grainy mouthfeel.
[0162] "Carriers" or "vehicles" indicate materials suitable for administering compounds and include any material known in the art, such as any liquid, gel, solvent, liquid diluent, solubilizer or the like, that is non-toxic and does not interact with no component of the composition in a harmful way.
[0163] Examples of nutritionally acceptable carriers include, for example, water, salt solutions, alcohol, silicone, waxes, petroleum jelly, vegetable oils, polyethylene glycols, propylene glycol, liposomes, sugars, gelatin, lactose, amylose, stearate magnesium, talc, surfactants, silicic acid, viscous paraffin, perfume oil, fatty acid monoglycerides and diglycerides, petroetral fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone and the like.
[0164] Examples of excipients include one or more of: microcrystalline cellulose and other celluloses, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, glycine, starch, milk sugar and high molecular weight polyethylene glycols.
[0165] Examples of disintegrants include one or more of: starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates.
[0166] Examples of granulation binders include one or more of: polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, maltose, gelatin and acacia.
[0167] Examples of lubricating agents include one or more of: magnesium stearate, stearic acid, glyceryl behenate and talc.
[0168] Examples of diluents include one or more of: water, ethanol, propylene glycol, glycerin and combinations thereof.
[0169] The other components can be used simultaneously (when they are, for example, in mixture with each other or even when they are supplied by different routes) or sequentially (when they can, for example, be supplied by different routes).
[0170] Preferably, when the composition according to the present invention is mixed with any other components, the microorganisms remain viable.
[0171] As used herein, the expression "component suitable for human or animal consumption" indicates a compound that is or can be added to the composition according to the present invention as a supplement that can be of nutritional benefit, a fiber substitute or has a generally beneficial effect on the consumer. The ingredients can be used in a wide variety of products that require gelling, texturing, stabilizing, suspending, film forming and structuring and juice retention, without unnecessary increase in viscosity. Preferably, the ingredients will be able to increase shelf life and stability of viable crop.
[0172] Components can be prebiotics such as alginate, xanthan, pectin, locust bean gamma (LBG), inulin, guar gum, galacto-oligosaccharide (GOS), fructo-oligosaccharide (FOS), polydextrose (ie , Litesse®), lactitol, lactosaccharose, soy oligosaccharides, palatinose, isomalto-oligosaccharides, gluco-oligosaccharides and xylo-oligosaccharides.
[0173] The ideal amount of the composition to be used in the combination according to the present invention will depend on the product to be treated and/or the method of contact of the product with the composition and/or the intended use for it. The amount of viable microorganism used in the compositions should be an amount sufficient to be effective and remain effective enough to improve aroma, taste, smoothness, consistency, texture, body, mouthfeel, viscosity, structure and/or organoleptic properties , nutrition and/or health benefits of food products containing said composition. This period of effectiveness should extend to at least the time of use of the product.
[0174] Concentrates: The compositions for use in the present invention may be in the form of concentrates. Typically, such concentrates comprise a substantially high concentration of Bifidobacterium lactis BL04 (such as viable Bifidobacterium lactis BL04) and/or its fermentation product and/or cell lysate.
[0175] Powders, granules and liquid compositions in the form of concentrates can be diluted with water or resuspended in water or other suitable diluents, such as a suitable growth medium such as milk or mineral or vegetable oils, to generate ready-to-use compositions .
[0176] The combinations according to the present invention in the form of concentrates can be prepared according to methods known in the art.
[0177] In one aspect of the present invention, the product is brought into contact with a composition in a concentrated form. Preferably, the product is brought into contact with a spray dried and/or resuspended composition.
[0178] The compositions according to the present invention can be spray dried or freeze dried by methods known in the art.
[0179] Typical particle elaboration processes using a spray drying process involve a solid material that is dissolved in an appropriate solvent (such as a microorganism cultivation in a fermentation medium). Alternatively, the material can be suspended or emulsified in non-solvent to form a suspension or emulsion. Other ingredients (as discussed above) or components such as antimicrobial agents, stabilizing agents, dyes and agents that assist in the drying process may optionally be added at this stage.
[0180] The solution is then atomized to form a fine mist of droplets. The droplets immediately enter a drying chamber, where they come into contact with a drying gas. The solvent is evaporated from the droplets into the drying gas to solidify the droplets to form particles. The particles are then separated from the drying gas and collected.
[0181] Products: Any product that can benefit from the composition can be used in the present invention. These include, but are not limited to, dairy foods and products derived from dairy foods, food supplements and pharmaceuticals.
[0182] By way of example, the composition according to the present invention can be used as an ingredient in soft drinks, fruit juices or drinks comprising whey protein, healthy teas, cocoa drinks, dairy drinks and acid bacteria drinks dairy, yogurt and yogurt drinks, cheese, ice cream, ice cream and desserts, confections, cookies, cakes and cake mixes, snacks, balanced drinks and foods, fruit fillings, toppings, bakery chocolate filling, flavored cake filling cheese, fruit flavored cake filling, topping for cakes and donuts, instant bakery filling creams, biscuit fillings, ready-to-use bakery filling, low-calorie fillings, nutritional drinks for adults, juice and soy drinks acidified, purified/aseptic chocolate drinks, bar mixes, powdered drinks, calcium fortified chocolate and whole/soy milk and calcium fortified coffee drink .
[0183] The composition can be additionally used as an ingredient in food products such as American cheese sauce, anti-caking agent for grated and grated cheese, snack sauce, cream cheese, fat-free sour cream for whipped and dry mixed topping, frozen/thawed sour cream, frozen/thawed stable whipped topping, natural light and low-fat cheddar cheese, low-fat Swiss yogurt, aerated frozen desserts, hard-packed ice cream, hard-packed ice cream with appropriate label , more economical and more indulgent, low-fat ice cream: to serve soft, barbecue sauce, dipping cheese sauce, cottage cheese topping, dry-mix Alfredo sauce, blended cheese sauce, tomato blend drought and others.
[0184] For certain aspects, the present invention can be preferably used in connection with the production of yogurt, such as fermented yogurt drinks, yogurt, drinking yogurt, cheese, fermented cream, milk-based desserts and others.
[0185] Suitably, the composition can be additionally used as an ingredient in one or more of cheese applications, meat applications or applications comprising protective crops.
[0186] The present invention also provides a method of preparing food or food ingredients, wherein the method comprises mixing the composition according to the present invention with another food ingredient.
[0187] Conveniently, the present invention relates to products that have been brought into contact with the composition according to the present invention (and optionally with other components/ingredients), wherein the composition is used in an amount that is able to enhance the nutrition and/or health benefits of the product.
[0188] As used herein, the term "put in contact" designates the direct or indirect application of the composition according to the present invention to the product. Examples of application methods that can be used include, but are not limited to, treating the product in a material comprising the composition, directly applying by mixing the composition with the product, spraying the composition onto the surface of the product, or dipping the product in a preparation of makeup.
[0189] When the product according to the present invention is a food, the composition according to the present invention is preferably mixed with the product. Alternatively, the composition can be included in the emulsion or starting ingredients of a food. In a further alternative, the composition can be applied in the form of a seasoning, coating, color mixture and the like.
[0190] For some applications, it is important that the composition is made available on the surface of a product to be affected/treated. This allows the composition to provide one or more of the following favorable characteristics: nutrition and/or health benefits.
[0191] The compositions according to the present invention can be applied to the intermediate space, coating and/or impregnating a product with a controlled amount of a viable microorganism.
[0192] Food: The composition according to the present invention can be used as food or in its preparation. The term “food” is used in a broad sense here and covers food for humans as well as food for animals (ie, feed). In a preferred aspect, the food is intended for human consumption.
[0193] The food can be presented in the form of a solution or solid, depending on the use and/or mode of application and/or mode of administration.
[0194] When used as a food, such as a functional food, or in its preparation, the composition according to the present invention may be employed in conjunction with one or more of: a nutritionally acceptable carrier, a nutritionally acceptable diluent, a nutritionally excipient acceptable, a nutritionally acceptable adjuvant or a nutritionally active ingredient.
[0195] Preferably, the composition is used to ferment milk, milk fortified with sucrose or lactic medium with sucrose and/or maltose, wherein the resulting medium containing all the components of the composition (i.e., the aforementioned microorganisms according to with the present invention) can be added as an ingredient to yoghurt milk at appropriate concentrations, such as concentrations in the final product offering a daily dose of 106 to 1010 cfu. The microorganism according to the present invention can be used before or after the yogurt fermentation.
[0196] For some aspects, the microorganisms according to the present invention or the composition according to the present invention are used as animal feeds, such as feed for farm animals, particularly feed from poultry (such as chickens) or pet food, or in its preparation.
[0197] Food ingredient: The composition according to the present invention can be used as a food ingredient and/or feed ingredient.
[0198] As used herein, the term "food ingredient" or "feed ingredient" includes a formulation that is or can be added to functional foods or feeds as a nutritional supplement.
[0199] The food ingredient can be in the form of solution or solid, depending on the use and/or mode of application and/or mode of administration.
[0200] Food supplements: The composition according to the present invention can be a food supplement or be added thereto.
[0201] Functional food: The composition according to the present invention can be a functional food or be added thereto.
[0202] As used herein, the expression "functional food" indicates a food that is able to provide not only nutritional effect, but is also able to provide additional beneficial effect to the consumer.
[0203] Consequently, functional foods are common foods that contain components or ingredients (such as those described herein) incorporated into them to provide the food with a specific function (such as medical or physiological benefit) other than a purely nutritional effect.
[0204] While there is no legal definition of a functional food, most parties with an interest in this area agree that they are foods marketed as having specific health effects beyond the basic nutritional effects.
[0205] Some functional foods are nutraceuticals. At present, the expression “nutraceutical” indicates a food that is capable of providing not only a nutritional effect and/or taste satisfaction, but is also capable of providing a therapeutic (or other beneficial) effect to the consumer. Nutraceuticals cross traditional boundaries between food and medicine.
[0206] Probiotics: For some applications, it is believed that the viable Bifidobacterium lactis BL04 in the composition according to the present invention can exert probiotic cultivation effect. It is also within the scope of the present invention to add additional probiotics and/or prebiotics to the composition according to the present invention.
[0207] At present, prebiotic is: "a non-digestible food ingredient that beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited amount of beneficial bacteria".
[0208] The term "probiotic cultivation", as used herein, defines live microorganisms (including, for example, bacteria or yeast) that, for example, when ingested or applied locally in sufficient quantities, beneficially affect the host organism , that is, conferring one or more demonstrable health benefits on the host organism. Probiotics can improve microbial balance on one or more mucosal surfaces. The mucosal surface can be, for example, the intestine, the urinary tract, the respiratory tract or the skin. The term "probiotic" as used herein also encompasses live microorganisms that can stimulate the beneficial branches of the immune system and, at the same time, reduce inflammatory reactions on the surface of the mucosa, such as the intestine.
[0209] Although there are no upper or lower limits of absorption of probiotics, it has been suggested that at least 106-1012, preferably at least 106-1010, preferably 108-109 cfu as a daily dose will be effective in achieving beneficial health effects in host organisms such as humans.
[0210] In addition to the probiotic effect, the microorganism according to the present invention can possess and is also within the scope of the present invention to provide prebiotics as other compounds that can be included in combination together with the composition. The prebiotic component of the combination comprising the composition according to the present invention is characterized by slow fermentation in the large intestine. These prebiotics can have a positive effect on the intestinal flora, specifically on the left side of the colon, an area of the intestine that is especially prone to disorders, particularly bowel cancer and ulcerative colitis.
[0211] Prebiotics are typically non-digestible carbohydrates (oligo or polysaccharides) or sugar alcohol that is neither degraded nor absorbed in the upper digestive tract. Known prebiotics used in commercial products and useful in accordance with the present invention include inulin (fructooligosaccharides, or FOS) and transgalactooligosaccharides (GOS or TOS). Other suitable prebiotics include palatino-oligosaccharide, soy oligosaccharide, gentio-oligosaccharide, xylo-oligomers, non-degradable starch, lactulose, lactulose, lactitol, maltitol, polydextrose (i.e., Litesse®) or the like.
[0212] In one embodiment, the present invention relates to the combination of Bifidobacterium lactis BL04 and/or its fermentation product and/or its cell lysate according to the present invention with a prebiotic.
[0213] The prebiotic may be administered simultaneously (such as in admixture or provided simultaneously by the same or different routes) or sequentially (such as by the same or different routes) with the microorganism hereunder invention and/or its fermentation product and/or its cell lysate.
[0214] Symbiotics: The present invention also contemplates the use of pre and probiotics as ingredients in a combination together with the composition according to the present invention which, when combined, become symbiotic. The purpose of this is to combine the effects of beneficial bacteria and the stimulation of the body's own beneficial bacteria. There is high potential in the development and consumption of these blends, as some of them may also exhibit powerful synergistic nutritional and/or health effects.
[0215] In this way, the composition according to the present invention can be specifically designed to contain different components that can provide symbiotic effect for the consumer.
[0216] Pharmaceuticals: The composition according to the present invention can be used as a pharmaceutical product or in its preparation. At present, the term “pharmaceutical product” is used in a broad sense and covers pharmaceuticals for humans as well as pharmaceuticals for animals (ie veterinary applications). In a preferred aspect, the pharmaceutical product is intended for human and/or animal mating use. The pharmaceutical product may be intended for therapeutic purposes, which may be curative, palliative or preventive in nature. The pharmaceutical product may even be intended for diagnostic purposes.
[0217] When used as a pharmaceutical product or in its preparation, the composition according to the present invention can be used in conjunction with one or more of: a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, a pharmaceutically acceptable excipient, a pharmaceutically adjuvant acceptable or a pharmaceutically active ingredient.
[0218] The pharmaceutical product may be in the form of a solution or solid, depending on the use and/or mode of application and/or mode of administration.
[0219] Pharmaceutical ingredient: The microorganisms according to the present invention can be used as pharmaceutical ingredients. At this point, the composition can be the only active component or it can be at least one of a series (that is, two or more) of active components.
[0220] The pharmaceutical ingredient may be in the form of a solution or solid, depending on the use and/or mode of application and/or mode of administration.
[0221] Forms: The microorganism according to the present invention and/or its fermentation product and/or its cell lysate can be used in any appropriate way, either alone or when present in a combination with other components or ingredients. Similarly, combinations comprising the composition according to the present invention and other components and/or ingredients (i.e. ingredients such as food ingredients, functional food ingredients or pharmaceutical ingredients) can be used in any suitable form.
[0222] The microorganism according to the present invention or the composition according to the present invention can be used in the form of solid or liquid preparations, or alternatives thereof. Examples of solid preparations include, but are not limited to, tablets, capsules, dusts, granules and powders which can be wettable, spray dried or freeze dried. Examples of liquid preparations include, but are not limited to, aqueous, organic or organic aqueous solutions, suspensions and emulsions.
[0223] Suitable examples of forms include one or more of: lozenges, pills, capsules, eggs, solutions or suspensions, which may contain flavoring or coloring agents, for immediate, delayed, modified, sustained, pulsed or controlled release applications.
[0224] As an example, if the composition according to the present invention is used in tablet form, such as for use as a functional ingredient, the tablets may also contain one or more of: excipients such as microcrystalline cellulose, lactose, citrate sodium, calcium carbonate, dibasic calcium phosphate and glycine; disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates; granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia; lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate and talc may be included.
[0225] Examples of nutritionally acceptable carriers for use in preparing the forms include, for example, water, salt solutions, alcohol, silicone, waxes, petroleum jelly, vegetable oils, polyethylene glycols, propylene glycol, liposomes, sugars, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oil, fatty acid monoglycerides and diglycerides, petroetral fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone and the like.
[0226] Preferred excipients for the forms include lactose, starch, cellulose, milk sugar or high molecular weight polyethylene glycols.
[0227] For aqueous suspensions and/or elixirs, the composition according to the present invention can be combined with various flavoring or sweetening agents, coloring material or dyes, with emulsifying and/or suspension-forming agents and with diluents such as water, propylene glycol and glycerin, as well as combinations thereof.
[0228] Forms may also include gelatin capsules; fiber capsules, fiber pellets etc.; or even fiber drinks.
[0229] Further examples of shape are found, for example, in cream form. For some aspects, the microorganism and/or one of its metabolites can be included in cosmetic and/or pharmaceutical creams such as sun creams and/or after sun creams, for example.
[0230] In one aspect, the composition according to the present invention can be administered in an aerosol, such as in the form of a nasal spray, for example, for administration to the respiratory tract. EXAMPLES
[0231] The present invention will be further described by reference to the following examples and figures, in which: - Figure 1 shows a schematic representation of the experiments performed to show the present invention; and - Figure 2 displays a Consort flowchart that details patient recruitment, processing, and analysis.
[0232] Methodology: Experimental part: The study involved a double-blind placebo-controlled trial of healthy physically active individuals from the community to establish whether 150 days of probiotic supplementation reduces URTI during the winter period between June and October 2010 (Figure 1 ). There were two experimental groups, a placebo group and a probiotic group, comprising 309 healthy physically active adults, 157 men and 152 women, in equivalent numbers. Participants completed a fourteen-day drive during which all use of probiotics and fortified foods/probiotic supplements and medications was discontinued. After baseline sampling, patients underwent a 150-day supplementation period. All participants were asked to keep a disease diary to record disease patterns (duration and severity). A set of participants from each group was selected for secondary analysis of immune and microbiological function. Each set of participants provided blood and stool samples and a throat swab to examine the effect of supplementation on URT and enteric microbiota, indices of innate immune function (NK cell function and phagocytosis). Saliva samples were collected, but the analysis will depend on whether or not beneficial clinical results are identified. Secondary analysis involved collecting fecal swabs from patients who traveled to Asia to determine whether probiotic supplementation reduces colonization of antibiotic-resistant Escherichia coli during the trip.
[0233] Granted ethics committee clearance by the Australian Institute of Sports Ethics Committee (February 19, 2010) and Griffith University (March 11, 2010).
[0234] Patients and recruitment: There were 268 healthy physically active community members recruited into the study. Of these, 226 individuals were included in the statistical analysis of physical activity and disease measures. Characteristics of patients included in the statistical analysis are detailed in Table 1. There were no substantial differences between groups.
[0235] Inclusion criteria: Inclusion in the study was determined according to levels of physical activity, in which participants needed to undergo at least three exercise sessions weekly.
[0236] Exclusion criteria: All participants were asked to declare their use of food aids and/or ergogenics that could influence the underlying immune function. All participants with immunomodulatory medications were excluded, including those with steroid-based asthma-fighting treatments. Patients who were on antibiotic treatments in the previous month were also excluded. Patients with any symptoms of gastrointestinal disease such as Crohn's disease, celiac disease and related conditions were excluded.
[0237] Primary outcome measures: The primary clinical outcome measure was URT disease in participants over the study period. Patients were asked to record any symptoms of URTI and chest disease in a diary of illness throughout the study period. Briefly, symptoms of URTI included sore throat, sneezing, stuffy nose or runny nose, and coughing. Symptoms of lower respiratory illness included coughing up chest congestion and/or sneezing. A classification of an episode of illness was performed when two or more symptoms were recorded on consecutive days. Functional impact or symptom severity for physically active individuals was self-rated as mild, moderate, or severe based on the impact of symptoms on daily activity for that day: mild, no change; moderate, reduced normal activity; and severe: complete termination of the activity.
[0238] Secondary outcome measures: Perceived tension and resilience: The tension and perceived resilience of the participants were measured using a questionnaire before and after supplementation. Social and psychological factors represent a source of tension that can affect immunity and health. All patients completed the Connor Davidson Resilience Questionnaire before and at the end of supplementation.
[0239] One set of participants (53 in the B. lactis BL-04 group, 51 in the placebo group) from each of the treatment groups provided samples for the following secondary outcome measures.
[0240] Faecal microbiology: - Total bacterial count (eubacteria): - Quantification of bacterial groups in faecal samples: These groups may include, but are not limited to, Bacteroidetes, Enterobacteriaceae, Lactobacillus spp, Bifidobacterium spp, Clostridium XIV set and other clostridial sets , Clostridium difficile, Collinsella, Escherichia coli, Enterococcus spp, Faecalibacterium prausnitzii, Roseburia spp, Veillonella spp and sulfate-reducing bacteria. Bacterial quantification will be conducted with qPCR and/or other relevant independent culture methods. Analysis of throat pad bacteria by qPCR will focus on particularly relevant bacterial groups, which may include, but are not limited to, Staphylococcus aureus, Pneumococcus spp and Streptococcus spp. - Antibiotic-resistant Escherichia coli: - L. acidophilus NCFM and B. lactis Bi-07 and B. lactis BL-04: Serum: - Natural killer cell activity: - Phagocytosis: Data analysis: A practical approach was used to perform inference (conclusion) about the clinical and physiological effects of probiotic treatments. This approach has been detailed in several articles (6). This approach is also consistent with the guidelines of the International Committee of Medical Publication Editors for determining clinical trials. The merits of this approach and some of the disadvantages of an approach based on testing hypotheses and statistical significance are well recognized. The approach is based on whether the uncertainty range of the true value of an effect is related to limits of values that are clinically important. The uncertainty of the true value is the confidence interval. As there are a large number of effects in this study, a conservative 99% confidence interval was selected; in other words, there is a 99% chance that the true value of each effect lies within the confidence interval that is calculated from the data. When the confidence interval includes values that are substantial in some positive and negative sense, such as beneficial and harmful, the effect can be both beneficial and detrimental and, therefore, the effect is deduced to be inconclusive or uncertain. Otherwise, it is deduced that the effect is clear and the magnitude attributed to the effect is the observed magnitude, such as beneficial, trivial, or harmful difference.
[0241] Thresholds selected as clinically important differ from the different types of variable outcomes in the study. For variables such as symptom intensity on a three-point scale, the effect of probiotic treatment was analyzed as a simple difference in means: probiotic indicates less than the placebo mean. For this type of effect, the standard limits are positive and negative difference in the mean equal to 0.20 of the standard deviation pooled between patients in both groups. This minor effects approach is known as standardization and provides moderate, large, and very large effect thresholds (0.60, 1.20, and 2.0 standard deviations). Other variables for which we selected magnitude limits in this way were the amount of medications taken per hundred days, the intensity of physical activity, the number of exercise days per week, total hours of exercise per week, total activity load per week (sum of the product of exercise intensity and the number of exercise days per week) and the variables from the saliva samples that were measured for their potential role as mechanisms of any treatment effect. Measures of training hours and training load were log-transformed before analysis to allow adequate analysis of the treatment effect as a percentage, but the magnitude of the effect was determined for the log-transformed variable. Saliva measurements were also log-transformed prior to analysis, but the variability and effects of these variables were generally much greater than for the training variables and therefore were expressed as factors.
[0242] The magnitude limits of variables that represent or involve the presence or symptom count needed to be determined differently, as the use of standard deviation for these variables is not appropriate. The variables in question were the number of episodes of a given symptom per one hundred days, the total number of symptom days per one hundred days, and the total symptom load per one hundred days (the sum of the product of the symptom intensity and the number of days of symptom for one hundred days). The effect of probiotic treatment on all these variables was analyzed as ratio: the mean of the probiotic group divided by the mean of the placebo group. We consider a ratio of 1.10 (ie, 10% higher mean value of the variable in the probiotic group) as the limit for substantial increase. For statistical reasons, the substantial reduction threshold in the probiotic group was therefore a ratio of 1/1.2 or 0.83. These ratios are similar to the risk of illness and injury ratios in public health studies. Epidemiologists consider that the risk ratio of 1.1 to 1.3 represents a substantial increase in risk. Unfortunately, so far, there is no consensus on limits that represent moderate, large and very large increases and decreases in risk.
[0243] Data from "shoulder" periods defined as two weeks after the start and end of supplementation were analyzed using a linear weighting factor to assign appropriate proportion of training and symptom ratings to baseline and treatment periods total. Thus, on the first day after starting treatment, 13/14 of the patient values were assigned to the baseline period and 1/14 was assigned to the treatment period. On the second day of the shoulder, the fractions were 12/14 and 2/14 and so on.
[0244] Baseline data were also taken into account. When recording baseline values, it is common to adjust for differences between patients at baseline by subtracting baseline ratings from treatment ratings. This strategy typically results in greater precision of the treatment effect than that provided by the analysis of treatment assessments alone, and therefore allows for the use of smaller sample sizes. It is generally not appreciated, however, that adjusting a baseline assessment in this way results in better accuracy only when the variable under analysis is reasonably reliable (ie, patient assessments tend to be consistently different from each other on prior and prior tests. after treatment). In this study, training variables and symptoms were analyzed as treatment-only assessments. It was clear that the effect of symptom treatment was more accurate for the treatment-only analysis, while the effect on training was more accurate for baseline treatment changes: evidently, patient training was more reliable and well defined by Baseline monitoring that your disease symptoms. The treatment-only analysis is therefore presented for the symptoms and baseline analyzes of the treatment for training. When making the decision to use treatment versus treatment-only baseline analyses, we deliberately avoided examining the magnitude of the effect and focused only on comparing the accuracy of the treatment effect estimate. Saliva variables were also presented as baseline treatment analyses.
[0245] The confidence limits of symptom assessments were estimated using an empirical method known as bootstrapping, as the usual analytical approaches involve hypotheses that are difficult to justify for measures involving the duration of the symptom. Bootstrapping was also used with training measures. These analyzes were performed using programs written in the Statistical Analysis System.
[0246] Treatments: Supplement Z contained a Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 combined; the dosage was 5x109 CFU/day for each bacterium and, therefore, with a total dosage of 109 CFU/day.
[0247] Supplement X contained Bifidobacterium lactis BL-04; the dosage was 2x109 CFU/day.
[0248] The placebo supplement Y contained sucrose.
[0249] Supplements Z and X were freeze-dried bacteria mixed in a cold drink (no alcohol would be consumed and the drink was not hot).
[0250] Results: A Consort flowchart that details the recruitment, processing, and analysis of patients is presented in Figure 2.
[0251] Patient details: Physical and physiological characteristics: The characteristics of patients included for analysis are detailed in Table 1 for each of the groups. There were no substantial differences in gender, age or body mass index between groups. TABLE 1
[0252] Characteristics of individuals included for analysis in each of the groups (mean ± standard deviation):

[0253] Care: Details of care and the average number of supplement days completed by participants in each group are described in Table 2. TABLE 2
[0254] Degree of care and average number of supplement days completed by participants in each of the three treatment groups over the study period:

[0255] Harmful effects: Four patients experienced diarrhea and cramping at the start of supplementation. Three of these patients withdrew and symptoms appeared in the third. One patient withdrew due to headaches that started with supplementation, including after a break in taking the assigned supplement. One patient suffered urticaria after starting supplementation and withdrew. One patient experienced intestinal pain after traveling to Asia. The travel date coincided with taking the supplement and the patient left.
[0256] Dietary information: Dietary information regarding fiber intake in each of the treatment groups is found in Table 3. There were no substantial differences in fiber servings between the groups. TABLE 3
[0257] Number of standard fiber servings per day per treatment group and sex at midpoint (Mid), end of study (End) and mean of two time points (Mean):

[0258] Pla: placebo.
[0259] Physical activity information: Details of participant training by treatment group during the study are presented in Table 4. There were no substantial differences in physical activity patterns between groups. TABLE 4
[0260] Training of patients during the study. Intensity rated on a scale from 1 to 10; training load is the sum of the product of training intensity and training duration in arbitrary units.


[0261] Disease episodes: Upper respiratory tract disease: The effect of probiotic supplementation on the number of respiratory tract disease episodes of variable duration is shown in Table 5a. B. lactis BL04 reduced the amount of longer-lasting respiratory tract diseases. Furthermore, this effect was more pronounced as the episodes of illness became longer. TABLE 5A
[0262] Effect of probiotic treatment on upper respiratory tract symptoms:

[0263] Chest infection (lower respiratory tract infection): The effect of B. lactis on chest infection is found in Table 5b. Similar to the effects of B. lactis BL04 on URTI, supplementation reduced the number of episodes of chest infection that lasted five days compared to episodes of shorter duration. TABLE 5b
[0264] Effect of probiotic treatment on chest symptoms:
DISEASE PATTERNS
[0265] The difference in frequency, duration, severity, and combined burden of upper respiratory disease between probiotic and placebo groups during the treatment period is shown in Table 6. There was substantially lower disease duration and burden among those taking BL- 04 compared to placebo. Based on sex, these reductions were also more pronounced in women than in men. TABLE 6
[0266] Effect of probiotic treatment on the amount, duration, severity, and combined burden of respiratory tract infections:


[0267] The difference in frequency, duration, severity and combined burden of breast disease (lower respiratory disease) between probiotic and placebo groups during the treatment period is shown in Table 7. There is a reduction in the severity of symptoms of breast infection. breast in BL-04 compared to placebo. TABLE 7
[0268] Effect of probiotic treatment on the amount, duration, severity, and combined burden of breast infections:



[0269] The difference in frequency, duration, severity and combined burden of medication use between groups during the treatment period is shown in Table 8. Briefly, participants with BL-04 had substantially total amount of medications and total days of medication lower than those on placebo. When examined by gender, the effect was maintained in men, but less so in women. TABLE 8
[0270] Effect of probiotic treatment on the amount, duration, severity, and combined burden of medication episodes:



[0271] Conclusions: The results demonstrate that supplementation with B. lactis BL-04 allowed a substantial reduction in the amount of upper and lower respiratory diseases and reduction in the severity and duration of URTI and lower respiratory tract symptoms.
[0272] Full data analysis: Provisional data analysis at 99% confidence interval suggested that B. lactis BL-04 may be effective in preventing respiratory tract infections and reducing the need for medications associated with respiratory infections (see Tables 5a, 5b, 6, 7 and 8). The encouraging interim results ensured complete data analysis, as shown below.
[0273] Complete data analysis (n = 399) was based on determining clinical significance as well as testing for statistical significance. Statistical significance was determined using traditional 95% confidence intervals. For the clinical significance test, the previously defined threshold of clinical relevance was set at 20% symptom reduction compared to the placebo group.
[0274] As shown in Table 9, treatment with B. lactis BI-04 markedly reduced symptoms of upper respiratory tract disease. The reducing effect was stronger for longer-lasting illnesses, that is, the most severe episodes of illness. Diseases lasting seven days or more were reduced by 46% compared to placebo. In all categories of disease duration (three days, five days and seven days or more), the reduction was greater than or equal to the previously defined cut-off value of clinical significance. Surprisingly, the effects of BL-04 on upper respiratory disease were stronger than for the combination of NCFM and Bi-07. This was particularly surprising because the BL-04 probiotic was given at a lower dose than the probiotic combination.
[0275] Both BL-04 and the combination of NCFM and Bi-07 were equally effective in reducing lower respiratory chest infections (see Table 9). Separate analysis also demonstrated that NCFM + Bi-07 combined, as well as BL-04 as a single strand, had a significant reducing effect on the duration of disease episodes.


[0276] Table 9 - Effect of probiotic supplements on the rate of upper and lower respiratory tract diseases and use of cold and flu medications, stratified by disease duration: Effects that exceed the previously defined clinical significance cut-off value (20 % reduction) are marked in bold. The BI-04 group showed greater reduction in disease velocity compared to placebo than the NCFM + Bi-07 group combined compared to placebo. REFERENCES 1. Albers, R., J.M. Antoine, R. Bourdet-Sicard, P.C. Calder, M. Gleeson, B. Lesourd, S. Samartin, I.R. Sanderson, J. Van Loo, F.W. Vas Dias and B. Watzl. Markers to measure immunomodulation in human nutrition intervention studies. Br J Nutr. 94: 452-481, 2005. 2. Berg, A., H.M. Muller, S. Rathmann and P. Deibert. The gastrointestinal system--an essential target organ of the athlete's health and physical performance. Exerc Immunol Rev. 5: 78-95, 1999. 3. Cox, A.J., D.B. Pyne, P.U. Saunders, R. Callister and M. Gleeson. Cytokine responses to treadmill running in healthy and illness-prone athletes. Med Sci Sports Exercise 39: 1918-1926, 2007. 4. Cox, A.J., D.B. Pyne, P.U. Saunders and P.A. Fricker. Oral administration of the probiotic Lactobacillus fermentum VRI-003 and mucosal immunity in endurance athletes. Br J Sports Med, 2008. 5. de Vrese, M. and J. Schrezenmeir. Probiotics and non-intestinal infectious conditions. Br J Nutr. 88 Supp. 1: S59-66, 2002. 6. de Vrese, M. and J. Schrezenmeir. Probiotics, Prebiotics, and Synbiotics. Adv Biochem Eng Biotechnol, 2008. 7. de Vrese M et al, Probiotic bacteria reduced duration and severity but not the incidence of common cold episodes in a double blind, randomized, controlled trial, Vaccine, 2006 Nov 10, vol. 24, no. 44-46, p. 6670 (Epub June 6, 2006). 8. de Vrese, M., P. Winkler, P. Rautenberg, T. Harder, C. Noah, C. Laue, S. Ott, J. Hampe, S. Schreiber, K. Heller, and J. Schrezenmeir. Effect of Lactobacillus gasseri PA 16/8, Bifidobacterium longum SP 07/3, B. bifidum MF 20/5 on common cold episodes: a double blind, randomized, controlled trial. Clin Nutr. 24: 481-491, 2005. 9. Dixon, R.E., Economic costs of respiratory tract infections in the United States, Am J Med, 1985, vol. 78, no. 6B, 45-51 10. Fricker, P.A., D.B. Pyne, P.U. Saunders, A.J. Cox, M. Gleeson and R.D. Telford. Influence of training loads on patterns of illness in elite distance runners. Clin J Sport Med. 15: 246-252, 2005. 11. Genton, L., W. van Gemert, C. Pichard and P. Soeters. Physiological functions should be considered as true end points of nutritional intervention studies. Proc Nutr Soc. 64: 285-296, 2005. 12. Gleeson, M., W.A. McDonald, D.B. Pyne, A.W. Cripps, J.L. Francis, P.A. Fricker and R.L. Clancy. Salivary IgA levels and infection risk in elite swimmers. Med Sci Sports Exercise 31: 67-73, 1999. 13. Gleeson, M., D.B. Pyne and R. Callister. Exercise effects on mucosal immunity and risk of upper respiratory illness. International Sports Medicine Journal. 4: 1-14, 2003. 14. Hashem M and Hall C B, Respiratory syncytial virus in healthy adults: the cost of a cold, Journal of Clinical Virology, 2003, vol 27, no. 1, p. 14-21. 15. Kang, W. and K.A. Kudsk. Is there evidence that the gut contributes to mucosal immunity in humans JPEN J Parenter Enteral Nutr. 31: 246-258, 2007. 16. Kekkonen, R.A., T.J. Vasankari, T. Vuorimaa, T. Haahtela, I. Julkunen and R. Korpela. The effect of probiotics on respiratory infections and gastrointestinal symptoms during training in marathon runners. Int J Sport Nutr Exerc Metab. 17: 352-363, 2007. 17. Koh, D., Y. Yong, V. Ng and S. E. Chia. Stress, mucosal immunity, upper respiratory tract infections, and sickness absence. J Occup Environ Med. 44: 987-988, 2002. 18. Leder K et al, A community-based study of respiratory episodes in Melbourne, Australia, Aust. N Z J Public Health, 2003, vol. 27, no. 4, p. 399-404. 19. Müns, G., P. Singer, F. Wolf and I. Rubinstein. Impaired nasal mucociliary clearance in long distance runners. Int J Sports Med. 16: 209-213, 1995. 20. Peters, H.P., W.R. De Vries, G.P. Vanberge-Henegouwen and L.M. Akkermans. Potential benefits and hazards of physical activity and exercise on the gastrointestinal tract. Gut. 48: 435-439, 2001. 21. Pregliasco F et al, A new chance of preventing winter diseases by the administration of synbiotic formulations, J Clin Gastroenterol., September 2008, vol. 42, sup. 3, part 2, S224-233 22. Pyne, D.B. and M. Gleeson. Effects of intensive exercise training on immunity in athletes. Int J Sports Med. 19 Suppl. 3: S183-191; Discussion S191-184, 1998. 23. Pyne, D.B., W.G. Hopkins, A.M. Batterham, M. Gleeson and P.A. Fricker. Characterizing the individual performance responses to mild illness in international swimmers. Br J Sports Med. 39: 752-756, 2005. 24. Pyne, D.B., W.A. McDonald, M. Gleeson, A. Flanagan, R.L. Clancy and P.A. Fricker. Mucosal immunity, respiratory illness, and competitive performance in elite swimmers. Med Sci Sports Exercise 33: 348-353, 2001. 25. Saavedra, J.M. and A. Tschernia. Human studies with probiotics and prebiotics: clinical implications. Br J Nutr. 87 Supp. 2: S241-246, 2002. 26. Welfare, 2006 Australian Institute of Health and Welfare 2006, Australia's Health 2006, Australia's health no. 10 Cat. No. AUS 73. Canberra: AIHW. Page 107. 27. West, N.P., D.B. Pyne, J.M. Kyd, G.M.C. Renshaw, P. Fricker and A.W. Cripps. The effect of exercise on innate mucosal immunity. British Journal of Sports Medicine, 2008. 28. West, N.P., D.B. Pyne, G. Renshaw and A.W. Cripps. Antimicrobial peptides and proteins, exercise and innate mucosal immunity. FEMS Immunol Med Microbiol. 48: 293-304, 2006. 29. Winkler P et al, Effect of a dietary supplement containing probiotic bacteria plus vitamins and minerals on common cold infections and cellular immune parameters, Int. J Clin. Pharmacol. Ther., July 2005, vol. 43, no. 7, pp. 318-326. 1/1

权利要求:
Claims (4)
[0001]
1. USE OF A COMPOSITION, comprising Bifidobacterium lactis BL04 and/or a fermentation product of Bifidobacterium lactis BL04 and/or a cell lysate of Bifidobacterium lactis BL04, characterized in that it is in the manufacture of a drug for the treatment or prophylaxis of a disease of the respiratory tract in a patient selected from tonsillitis, otitis media; rhinitis; rhinosinusitis, sinusitis; nasopharyngitis, rhinopharyngitis, common cold; pharyngitis; epiglottitis; supraglottitis; laryngitis; laryngotracheitis; tracheitis; bronchitis; acute bronchitis; and pneumonia.
[0002]
2. USE according to claim 1, characterized in that the phagocytic activity of granulocytes in the patient is increased.
[0003]
3. USE according to any one of claims 1 to 2, characterized in that the phagocytic activity of monocytes in the patient is increased.
[0004]
4. USE according to any one of claims 1 to 3, characterized in that the use is prophylactic.

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WO2017028813A1|2015-08-19|2017-02-23|Lin Xu|A composition for reducing lung and/or systemic inflammation associated with pm2.5 exposure and the use thereof|

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CN112569262A|2015-11-20|2021-03-30|4D制药研究有限公司|Compositions comprising bacterial strains|

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GB201520497D0|2015-11-20|2016-01-06|4D Pharma Res Ltd|Compositions comprising bacterial strains|

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GB201520638D0|2015-11-23|2016-01-06|4D Pharma Res Ltd|Compositions comprising bacterial strains|

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JP6441536B2|2016-03-04|2018-12-19|フォーディー ファーマ ピーエルシー４Ｄ Ｐｈａｒｍａ Ｐｌｃ|Composition comprising a bacterial strain|

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TW201821093A|2016-07-13|2018-06-16|英商４Ｄ製藥有限公司|Compositions comprising bacterial strains|

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GB201612191D0|2016-07-13|2016-08-24|4D Pharma Plc|Compositions comprising bacterial strains|

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GB201621123D0|2016-12-12|2017-01-25|4D Pharma Plc|Compositions comprising bacterial strains|

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KR20190126309A|2017-01-31|2019-11-11|캔사스 스테이트 유니버시티 리서치 파운데이션|Microbial cells, methods of production thereof, and uses thereof|

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JP2020520911A|2017-05-22|2020-07-16|フォーディー ファーマ リサーチ リミテッド４Ｄ Ｐｈａｒｍａ Ｒｅｓｅａｒｃｈ Ｌｉｍｉｔｅｄ|Composition comprising a bacterial strain|

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WO2018229188A1|2017-06-14|2018-12-20|4D Pharma Research Limited|Compositions comprising bacterial strains|

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JPWO2019151371A1|2018-01-31|2021-01-07|ニュートリー株式会社|Preventive and / or therapeutic agents for pneumococcal infections|

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CN110055185B|2018-12-25|2020-12-29|江南大学|Bifidobacterium breve with anti-influenza capacity and application thereof|

法律状态:
2018-01-23| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|

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2018-04-03| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2019-03-19| B07E| Notification of approval relating to section 229 industrial property law [chapter 7.5 patent gazette]|Free format text: NOTIFICACAO DE ANUENCIA RELACIONADA COM O ART 229 DA LPI |

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2019-08-27| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2021-03-16| B06A| Patent application procedure suspended [chapter 6.1 patent gazette]|

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2021-07-13| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2021-08-31| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 08/06/2012, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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申请号 | 申请日 | 专利标题

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US201161495614P| true| 2011-06-10|2011-06-10|

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US61/495,614|2011-06-10|

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GBGB1110095.5A|GB201110095D0|2011-06-15|2011-06-15|Method of treatment|

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GB1110095.5|2011-06-15|

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PCT/GB2012/051302|WO2012168732A1|2011-06-10|2012-06-08|Treatment of respiratory tract illness with bifidobacterium lactis bl-04|

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