Composition comprising American acid fruit vine extract and phospholipid

Abstract

The present invention relates to a composition comprising an extract of American fruit tendril in combination with a phospholipid and its use in the prevention and treatment of urinary tract infections. The invention also relates to a method for preparing said composition, and to a formulation for oral administration comprising said composition.

Description

[0001] This application is a divisional application of the international application with the application number PCT / IB2019 / 056261, the title of which is “Compositions comprising American cranberry extract and phospholipids”, filed on July 23, 2019, which entered the Chinese national phase on January 22, 2021, and the application number is 201980049079.8. TECHNICAL FIELD

[0002] The present application relates to a pharmaceutical or nutritional composition comprising a plant extract, in particular to a composition for the treatment and prevention of urinary tract bacterial infections. BACKGROUND

[0003] Juices and extracts of American cranberry (Vaccinium macrocarpon Aiton) have been the subject of a large number of clinical trials for the treatment and prevention of lower urinary tract infections (Burleigh A.E. et al. Nutr J 2013; 12: 139, Ledda A. et al. Eur Rev Med Pharmacol Sci 2017; 21: 389-393, Jepson RG. et al. Cochrane Database Syst Rev 2012; 10: CD001321). Recurrent infections of the lower urinary tract affect about 50% of adult women (Wang A. et al. Prim Care 2013; 40: 687-706) and can also affect men, although less frequently. In men, the incidence of lower urinary tract infections increases significantly with age, due to the structural and functional changes that occur in the urinary tract as a result of aging. The changes are usually manifested as a condition known as benign prostatic hyperplasia (Schaeffer AJ. et al. N Engl J Med 2016; 374: 562-571).

[0004] The main pathogen of lower urinary tract infections is the bacterium Escherichia coli, followed by Proteus spp., Staphylococcus saprophyticus, Klebsiella spp. and other Enterobacteriaceae (Kahlmeter G.J Antimicrob Chemother 2000; 46: 15-22).

[0005] Lower urinary tract infections are classified as recurrent if they occur at least 3 times in a year or 2 times in six months. Antibiotics can be used to prevent reducing the frequency of the episodes, but long-term antibiotic therapy increases health costs, can create resistance to the efficacy of antibacterial therapy and alter the intestinal flora.

[0006] Therefore, the identification of a non-antibiotic treatment that prevents the episodes of recurrent lower urinary tract infections would cause great interest.

[0007] A possible non-antibiotic treatment includes dietary supplements containing American cranberry extract, which has attracted a lot of interest. American cranberry contains many organic substances, including proanthocyanidins (PACs), flavonols, polyphenols and hydroxycinnamic acids, which, as well as their active metabolites, have direct and indirect bactericidal activity. The latter is attributed to their ability to prevent adhesion to the urothelial mucosa of the urinary tract, thus preventing the pathogen from forming a biofilm. The anti-adhesion activity has been demonstrated, in particular for proanthocyanidins - the main active ingredient of American cranberry. In vitro and in vivo tests have demonstrated that proanthocyanidins inhibit the adhesion of P fimbriae of E. coli to the urothelial mucosa of the urinary tract (Howell AB. et al. N Engl J Med 1998; 339: 1085-1086, Di Martino P. et al. J Urol 2006; 24: 21-27).

[0008] For example, Kakkar, Sahil et al. ISRN pharmacology 2014 (2014), Maria, et al. Molecules 20.8 (2015): 13536-13549 and Jayaraman, Premkumar, et al. International journal of biological sciences 6.6 (2010): 556 - Mensah, J.K. et al. Current Science Perspectives 1.2 (2015): 69-76) describe the antibacterial activity of some of the main metabolites present in the active ingredients of American cranberry extract, such as protocatechuic acid and gallic acid, on the strains responsible for lower urinary tract infections.

[0009] However, the efficacy data obtained so far on American cranberry derivatives in the treatment and prevention of urinary tract infections are somewhat erratic. This is partly due to the use of extracts that are neither well characterized nor reproducible, and that have too low a content of active ingredients, and especially to the use of compositions and formulations that are not suitable to promote the absorption of the active ingredients after oral administration, and especially to not allow the active metabolites to reach effective concentrations in the urine.

[0010] Therefore, the availability of compositions that promote the absorption of the active ingredients of American cranberry and of their metabolites after oral administration and ensure that they reach effective concentrations in the urine can represent an important tool as an alternative and complement to antibiotic therapy in the treatment and prevention of urinary tract infections.

[0011] For example, compositions obtained from the combination of plant extracts with natural or synthetic phospholipids are described in EP 0209037 (IdB Holding S.p.A.), EP 0275005 (Indena S.p.A.), EP 0283713 (Indena S.p.A.) and WO 2007 / 118631 (Indena S.p.A.).

[0012] WO 2010 / 150051 (Lipoid GmbH) describes compositions containing active plant ingredients or plant extracts, sugars, preferably maltodextrin, and phospholipids. The compositions are obtained by dissolving or dispersing the ingredients simultaneously or sequentially in water at room temperature. This step is followed by a homogenization, filtration and drying process, and optionally grinding to the desired particle size. In particular, WO 2010 / 150051 claims an active plant ingredient content of 0.5-40% by weight, a maltodextrin content of 10-90% by weight and a phospholipid content of 0.5-30% by weight. The document states that the compositions are characterized by high stability during transport and storage and by ease of incorporation into various dosage forms of the pharmaceutical, nutritional and cosmetic industries. The document also states that the compositions are characterized by good solubility and wettability, which ensures high bioavailability of the active plant ingredients. The active plant ingredients cited in WO 2010 / 150051 include American cranberry extract (Vaccinium macrocarpon). However, no composition containing American cranberry extract is specifically described. Moreover, no in vivo data are cited to demonstrate the actual increase in bioavailability of the active ingredients contained in the compositions, in particular no in vivo data are cited to demonstrate that the active metabolites reach effective concentrations in the urine excreted after oral administration, which is a necessary condition for the therapeutic and preventive action in the treatment and prevention of urinary tract infections.

[0013] Therefore, there is still a need to provide a cranberry-based composition ensuring a measurable effective level of active ingredients, in particular of their metabolites, in urine. SUMMARY

[0014] The present invention relates to a composition, typically in solid form, comprising:

[0015] a) an extract of Vaccinium macrocarpon, in an amount of 20% to 50% by weight of the total weight of the composition; and

[0016] b) at least one phospholipid, in an amount of 50% to 80% by weight of the total weight of the composition.

[0017] The present invention also relates to a process for the preparation of said composition as well as to a pharmaceutical or nutraceutical formulation for oral administration comprising said composition. The present invention also relates to a composition as described above and to the corresponding formulation for the treatment and prevention of urinary tract infections. BRIEF DESCRIPTION OF DRAWINGS

[0018] Fig. 1 shows the area under the curve (AUC) of epicatechin; Figures 1-4 Fig. 2 shows the area under the curve (AUC) of myricetin-3-O-glucoside;

[0019] Figure 1 Fig. 3 shows the area under the curve (AUC) of myricetin-3-β-O-galactoside;

[0020] Figure 2 Fig. 4 shows the area under the curve (AUC) of catechin-3-O-gallate.

[0021] Figure 3 Fig. 5 shows the area under the curve (AUC) of epicatechin gallate.

[0022] Figure 4 Fig. 6 shows the area under the curve (AUC) of quercetin. DETAILED DESCRIPTION

[0023] It has surprisingly been found that by combining an extract of Vaccinium macrocarpon, preferably an extract characterized by a high content of proanthocyanidins (PACs), with a high percentage of phospholipids (PLs), in particular by the preparation process described below, it is possible to reach in vivo a measurable effective urinary concentration of active metabolites of the active ingredients contained in the extract. In particular, it has been found that the composition of the present invention can be administered in a smaller amount compared to a Vaccinium macrocarpon extract having the same content of proanthocyanidins but not combined with phospholipids, while still ensuring that an effective concentration is reached in urine, and therefore in contact with the uroepithelial mucosa.

[0024] It has also been surprisingly found that, after administration of the formulations of the present application, the main metabolite appears in the urine at a different time, in particular later than after administration of a control formulation containing the "as is" extract, i.e. not combined with phospholipids. Therefore, the present application also relates to formulations containing the American cranberry extract combined with phospholipids and formulations of the American cranberry extract "as is", i.e. not combined with phospholipids.

[0025] The composition of the present application contains:

[0026] a) 20% - 50% (w / w) of American cranberry extract, preferably characterized by a proanthocyanidin content of 15% - 40% (w / w);

[0027] b) 50% - 80% (w / w) of phospholipids or a mixture of phospholipids.

[0028] The composition of the present application can optionally contain 1% - 30% by weight, relative to the total weight of the composition, of one or more technological adjuvants, such as diluents, glidants or lubricants, which improve the physical properties of the composition and facilitate their incorporation into the formulation.

[0029] In a preferred aspect, the composition consists of the above mentioned amounts of American cranberry extract and phospholipids or a mixture of phospholipids, corresponding to a weight ratio between extract and phospholipids of 1 : 1 to 1 : 4. In another preferred aspect, the composition consists of the above mentioned amounts of American cranberry extract, phospholipids or a mixture of phospholipids and one or more adjuvants. The proportions and the American cranberry extract and phospholipids under the reported conditions can be in the form of molecular complexes following the interaction between the chemical functions of the extract components and the chemical functions of the phospholipids.

[0030] For the purposes of the present application, the expression "American cranberry extract" means an extract obtainable by organic extraction methods from the pulp of Vaccinium macrocarpon Ait., preferably from the juice obtained by crushing them. The extract can be subsequently concentrated to increase the content of proanthocyanidins, which are considered the main (although not the only) active ingredient of this biomass. The extract has preferably a proanthocyanidin content of 15% - 40% (w / w), more preferably 25% - 35% (w / w), measured by the DMAC (4-dimethylamino cinnamaldehyde) colorimetric method (R.L. Prior et al., J Sci Food Agric 2010; 90; 1473-1478).

[0031] A preferred example of cranberry extract with high proanthocyanidin content is the one available on the market from Indena S.p.A. under the trade name Cran-Max®.

[0032] For the purposes of the present invention, the term "phospholipids" means lecithins, which can be obtained from soy, sunflower or other vegetable or animal sources, preferably vegetable sources, preferably lecithins selected from phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine, which have identical or different acyl groups, mainly palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid derivatives. The term "mixture" in relation to phospholipids means a mixture of lecithins derived from the same vegetable source or from different vegetable sources. The expression "at least one phospholipid" means a single phospholipid or a mixture of phospholipids.

[0033] The adjuvants that can be included in the composition of the present invention are typically selected from one or more of the following: - diluents, which are water-soluble or water-insoluble at room temperature, such as microcrystalline cellulose, calcium phosphate, calcium carbonate,

[0034] mannitol, maltodextrin, isomalt or combinations thereof;

[0035] - lubricants and / or glidants, such as silicon dioxide, talc, stearic acid, magnesium stearate or combinations thereof; - surfactants, such as sucrose esters, polysorbates, polyoxyethylene castor oil derivatives, D-alpha-tocopheryl-polyethylene glycol succinate (vitamin E TPGS) or combinations thereof.

[0036] These excipients and further excipients are described in Remington: "The Science and Practice of Pharmacy", 22ndedition, Pharmaceutical Press, 2013.

[0037] In a preferred aspect, the composition does not contain sugars, in particular does not contain maltodextrin, mannitol, isomalt or combinations thereof.

[0038] In a second aspect, the present invention relates to a process for obtaining said composition and to the composition obtainable by said process.

[0039] In particular, the process comprises dissolving or dispersing the American cranberry extract and at least one phospholipid and, optionally, at least one adjuvant in an organic solvent suitable for pharmaceutical use and / or nutritional use at a temperature of from 30°C to 90°C, preferably from 50°C to 90°C, obtaining a solution or dispersion, from which the organic solvent is subsequently removed, obtaining the solid composition of the present invention, said organic solvent being preferably selected from ethanol, ethyl acetate and acetone.

[0040] In a preferred aspect thereof, the process comprises simultaneously dissolving or dispersing the American cranberry extract and at least one phospholipid in an organic solvent, then optionally adding one or more adjuvants. According to said aspect, the process comprises the following steps:

[0041] a-1) dissolving or dispersing the extract and the phospholipid in 10-20 volumes of an organic solvent, preferably selected from ethanol, ethyl acetate and acetone, until a solution or a homogeneous dispersion is obtained;

[0042] b-1) heating the solution / dispersion obtained in step a) at a temperature ranging from 30°C to 90°C, preferably from 50°C to 90°C, typically maintaining the system under stirring until a solution or a homogeneous dispersion is obtained. The heating is necessary to maximize the solubility of the extract and the phospholipid;

[0043] c-1) optionally adding one or more excipients to the solution or dispersion in step b-1), while maintaining stirring under the same temperature conditions as in step b), until a solution or a homogeneous dispersion is obtained;

[0044] d-1) removing the solvent from the solution or dispersion obtained in step b) or c), preferably by evaporation at low pressure, more preferably at 10-60 MPA, while maintaining the temperature preferably at 50°C-70°C, more preferably at 70°C, optionally by vacuum drying in a stove set at 50°C-90°C, until a solid composition is obtained, which typically has a solvent residue lower than the limits specified in ICH Guideline Q3C (R6) concerning residual solvents;

[0045]

[0046] e-1) sieving the solid composition obtained at the end of step d) with a mesh of 0.25-2.0 mm, and optionally adding a lubricant and / or a glidant, preferably selected from stearic acid, magnesium stearate and silicon dioxide, preferably silicon dioxide.

[0047] In another preferred aspect thereof, the process comprises dissolving or dispersing the extract in a solvent until a solution or a dispersion is obtained, and adding at least one phospholipid to the solution or dispersion and optionally adding one or more excipients. In particular, according to said aspect, the process comprises:

[0048] a-2) dissolving or dispersing the American cranberry extract or the phospholipid in an organic solvent until a homogeneous solution or dispersion is obtained;

[0049] b-2) adding the American cranberry extract or the phospholipid to the obtained solution or dispersion, heating the mixture and maintaining stirring under the experimental conditions described in paragraph b);

[0050] - carrying out steps c-1), d-1) and e-1) as described above, until the process is completed.

[0051] ​For the avoidance of doubt, for the purposes of the present application, the term "solution" means a liquid composition which is clear in appearance upon visual inspection. The term "dispersion" means a liquid composition which contains suspended particles, is opaque and turbid in appearance upon visual inspection, but is still homogeneous.

[0052] Furthermore,

[0053] - where numerical ranges are given, it should be considered that the end values of the range are also included;

[0054] - the terms "process" and "method" are synonymous;

[0055] - the expressions "at least one" and "one or more" are equivalent and can be used interchangeably;

[0056] In its third aspect, the present application relates to a solid formulation for oral administration comprising the composition of the present application in admixture with one or more pharmaceutically or nutraceutically acceptable excipients. Examples of formulations include granules, tablets and capsules, optionally suitable for modified release, which can be prepared by the person skilled in the art by known methods, with known excipients such as those described in Remington: "The Science and Practice of Pharmacy", 22ndedition, Pharmaceutical Press, 2013. The formulation typically comprises 20-50% w / w of the amount of the formulation with respect to the total weight of the formulation, in such a way as to ensure a content of extract of 30-80 mg and an intake of proanthocyanidins of 9-25 mg per dose. According to a preferred embodiment, in addition to the composition of the present application, the formulation preferably also comprises the composition obtainable by the methods described above and the extract "as is", i.e. not in combination with phospholipids.

[0057] In its fourth aspect, the present application relates to a method for the prevention or treatment of urinary tract infections, in particular lower urinary tract infections, comprising administering to a patient in need thereof a composition or a formulation, preferably a formulation. The method preferably comprises administering an amount of the composition of the present application or preferably administering an amount of the formulation to ensure an intake of proanthocyanidins of 9-25 mg per day.

[0058] The following examples illustrate the present application in greater detail.

[0059] Experimental part

[0060] Materials

[0061] American cranberry extract (Vaccinium macrocarpon) was commercially available from Indena S.p.A. under the trade name Cran-Max®

[0062] Soy lecithin or sunflower lecithin was obtained from Cargill. Maltodextrin used in Example 3 was obtained from Roquette. Silicon dioxide used in Example 3 was obtained from Grace GmbH & Co. Microcrystalline cellulose used in Examples 4 and 5 was obtained from FMC Biopolymer, magnesium stearate used in Examples 4 and 5 was obtained from Peter Greven, dicalcium phosphate dihydrate used in Example 5 was obtained from Budenheim, polyvinylpyrrolidone used in Example 5 was obtained from Ashland.

[0063] In Examples 1-3, the percentages are weight percentages relative to the total weight of the composition.

[0064] Preparation Examples

[0065] Example 1 - Composition containing American cranberry extract and soy lecithin (C1)

[0066] American cranberry extract (Vaccinium macrocarpon) 25.0%

[0067] Soy lecithin 75.0%

[0068] The composition was obtained by the following method:

[0069] 1. The extract and the soy lecithin were dispersed in ethyl acetate saturated with 10 volumes of water relative to the total weight of the extract and the lecithin, under magnetic stirring.

[0070] 2. The dispersion obtained in step 1 was heated to reflux (about 70°C) and kept under stirring at said conditions for 3 hours.

[0071] 3. The solvent was removed from the dispersion obtained in step 2 by low pressure evaporation, while maintaining the temperature at < 70°C.

[0072] 4. Drying of the product obtained in step 3 was completed in an oven at 50°C under vacuum until obtaining ethyl acetate residues complying with the limits specified in ICH Guideline Q3C (R6).

[0073] 5. The product obtained in step 4 was calibrated with a 60 mesh sieve.

[0074] Example 2 - Composition containing American cranberry extract and soy lecithin (C2)

[0075] American cranberry extract (Vaccinium macrocarpon) 33.3%

[0076] Soy lecithin 66.7%

[0077] The composition is obtained by the following method:

[0078] 1. Soy lecithin is suspended in 15 volumes of technical ethanol under magnetic stirring, heated to 50°C until a dispersion is obtained.

[0079] 2. American cranberry extract is added to the dispersion obtained in step 1 under magnetic stirring. The dispersion is heated to reflux (about 70°C) and kept under stirring at said conditions for 2 hours.

[0080] 3. The solvent is removed from the dispersion obtained in step 3 by low pressure evaporation, while maintaining the temperature at < 70°C.

[0081] 4. Drying of the product obtained in step 3 is completed in an oven under vacuum at 50°C until the solvent residues in accordance with the limits specified in ICH Guideline Q3C (R6) are obtained.

[0082] 5. The product obtained in step 4 is calibrated with a 20-mesh sieve.

[0083] Example 3 - Composition containing American cranberry extract, sunflower lecithin, maltodextrin and silicon dioxide (C3) Example 4 - Formulation in the form of hard capsules containing composition (C1) (F1)

[0084]

[0085] The composition is obtained by the following method:

[0086] 1. The extract is dispersed in 10 volumes of water-saturated ethyl acetate under magnetic stirring.

[0087] 2. Lecithin is added to the dispersion obtained in step 1 under magnetic stirring.

[0088] 3. The dispersion is heated to reflux (about 70°C) and kept under stirring at said conditions for 2 hours.

[0089] 4. Maltodextrin is added to the dispersion obtained in step 3, while maintaining the same stirring and temperature conditions as described in step 3 for 30 minutes.

[0090] 5. The solvent is removed from the dispersion obtained in step 4 by low pressure evaporation, while maintaining the temperature at < 70°C.

[0091] 6. Drying of the product obtained in step 5 is completed in an oven under vacuum at 50°C until the solvent residues in accordance with the limits specified in ICH Guideline Q3C (R6) are obtained.

[0092] 7. The product obtained in step 6 is calibrated with a 20-mesh sieve.

[0093] 8. Silica is added to the product obtained in step 7, in an amount equal to 2% of the mixture, and mixed in a V-blender (MultiGel) for about 2 minutes.

[0094] Example 5 - Formulation in the form of tablets containing composition (C1) (F2)

[0095]

[0096] Formulation (F1) is obtained by the following method:

[0097] 1. Composition (C1) is mixed with microcrystalline cellulose in a V-blender (MultiGel) for about 5 minutes.

[0098] 2. Silica is added to the mixture obtained in step 1 and mixing is continued for about 2 minutes.

[0099] 3. Magnesium stearate is added to the mixture obtained in step 2 and mixing is continued for about 1 minute.

[0100] 4. The mixture obtained in step 3 is divided into hard or plant-based capsules at a rate of 260 mg / capsule.

[0101] Example 6 - Quantification of the active ingredient and active metabolites of the formulation containing composition (C1) in urine

[0102]

[0103] Formulation (F1) is obtained by the following method:

[0104] 1. Composition (C1) is mixed with calcium hydrogen phosphate dihydrate, microcrystalline cellulose and polyvinylpyrrolidone in a V-blender (MultiGel) for about 10 minutes.

[0105] 2. Silica is added to the mixture obtained in step 1 and mixing is continued for about 2 minutes.

[0106] 3. Magnesium stearate is added to the mixture obtained in step 2 and mixing is continued for about 1 minute.

[0107] 4. The mixture obtained in step 3 is pressed in a rotary tablet press equipped with a circular concave punch with a diameter of 12 mm.

[0108] In vivo test

[0109] Figures 1-4

[0110] The study was performed as follows: healthy volunteers (non-smoking women with a low polyphenol diet) were administered for one week, two capsules per day, each containing 120 mg of American cranberry extract not associated with phospholipids, the extract containing 30% proanthocyanidins, equivalent to 36 mg of proanthocyanidins [control capsules (F0)]. After a washout period of one week, the same volunteers were again administered for one week, two capsules per day, of a formulation (F1) prepared according to Example 4, each containing 120 mg of composition (C1), equivalent to 30 mg of American cranberry extract containing 30% proanthocyanidins. Urine was collected before the first administration and 1, 2, 4, 6, 10, 12 and 24 hours after the last administration of the control capsules (F0) and of the capsules of the formulation (F1). In order to identify and quantify the analytes related to the administered products in the urine, the collected urine samples were diluted 1:5 v / v with a 1% solution of trichloroacetic acid and then extracted with HyperSep C18 cartridges (5 ml) according to the manufacturer's instructions. The extracted metabolites present in the urine were then subjected to acid and enzymatic hydrolysis and then quantified by HPLC-mass spectrometry, using a Zorbax SB-C18 column for separation and a triple quadrupole mass spectrometer (TSQ Quantum Ultra AM®) as detector. TM C18 column solid phase extraction. The extracted metabolites present in the urine were then subjected to acid and enzymatic hydrolysis and then quantified by HPLC-mass spectrometry, using a Zorbax SB-C18 column for separation and a triple quadrupole mass spectrometer (TSQ Quantum TM Triple Quadrupole) as detector.

[0111] Surprisingly, it was found that, despite the fact that the amount of American cranberry extract, and therefore of proanthocyanidins, administered with the formulation (F1) containing 120 mg of composition (C1) was four times lower than the amount administered using capsules containing the "as is" extract [control capsules (F0)], some metabolites from proanthocyanidins and from other active ingredients of the extract (flavonols and phenolic acids) were more abundant after administration of the formulation (F1). In particular, the following metabolites were more abundant with respect to the amount of extract used: epicatechin, catechin-3-O-gallate, myricetin-3-beta-O-galactoside, myricetin-3-O-glucoside, quercetin-3-O-ramnoside, isorhamnetin-3-O-glucoside, kaempferol-3-O-glucoside, gallic acid, hippuric acid and protocatechuic acid.

[0112] The histograms demonstrating the experimental observations are shown in the accompanying Figure 1 figures, which give the area under the curve (AUC) for the four main components of the American cranberry extract: epicatechin ( Figure 2 ), myricetin-3-O-glucoside ( Figure 3 ), myricetin-3-beta-O-galactoside ( Figure 4 ) and catechin-3-O-gallate ( Example 7 - Efficacy study of the formulation containing composition (C1) in inhibiting the adhesion and biofilm formation of various strains of Candida albicans). The term "Cranberry" refers to the data obtained with the administration of the control capsules (F0) while the term Phytosome refers to the data obtained with the administration of the capsules containing the composition (C1).

[0113] The data demonstrate that, unlike the cranberry extract not associated with phospholipids, the composition of the application ensures that the main metabolite of the active ingredient of the extract, which reaches measurable effective levels in the urine, thus in contact with the urothelial mucosa, represents a substrate for bacterial infections.

[0114] It was also surprisingly found that the various metabolites appear in the urine at different times depending on whether the administration is of the control formulation (F0) containing the "as is" cranberry extract or of the formulation (F1) of the application, as clearly shown in the histograms below, in which the values corresponding to the administration of the control formulation (F0) are labelled "Cranberry" while the values corresponding to the administration of the formulation (F1) of the application are labelled "Phytosome".

[0115] This finding indicates that, by combining the "as is" (i.e. not associated with phospholipids) extract with the composition of the application, it is possible to ensure a persistent antibacterial action in the urothelial mucosa of the lower urinary tract, thus ensuring a more effective prophylactic action against said recurrent infections.

[0116] ​ ​

[0117] The efficacy of the formulation containing the composition (C1) was evaluated in an in vitro test, in particular the inhibitory effect on the adhesion of cells and on the biofilm formation of various strains of Candida albicans inoculated from vaginal swabs and of cell cultures from the reference strain of Candida albicans (SC5314). The degree of biofilm formation was quantified by staining with gentian violet and subsequent spectrophotometric reading at 540 nm.

[0118] A healthy female volunteer was treated daily for one week with 2 capsules each containing 120 mg of American cranberry extract not associated with phospholipids containing 30% proanthocyanidins, equivalent to 36 mg of proanthocyanidins [control capsules (F0)]. The cell cultures with biofilm were then brought into contact with the urine of the volunteer, which was freeze-dried and reconstituted with the same medium (RPMI-1640) used for the cell cultures. After a washout period of one week, the same healthy volunteer was treated daily for one week with 2 capsules of the formulation (F1) each containing 120 mg of the composition (C1) (equivalent to 30 mg of American cranberry extract containing 30% proanthocyanidins). In both cases, the urine was collected at 1, 2, 4, 6, 10, 12 and 24 hours after the intake of the last capsule required for the treatment.

[0119] Table 1 below gives absorbance data relating to spectrophotometer readings at 540 nm, which quantifies the extent of biofilm present on the cell culture. The values U pre(F0) and U pre(F1) refer to urine readings before administration.

[0120]

[0121] The comparison of the values shows that the inhibitory activity is clearly shown in the urine samples taken after 1 hour and 12 hours in the case of treatment with 120 mg of American cranberry [control capsule (F0)] and in the urine samples taken after 2 hours and 12 hours in the case of subjects treated with the capsules of the preparation (F1). Surprisingly, it was found that the inhibitory effect at the times indicated above is comparable, despite the fact that the preparation (F1) contains 4 times less American cranberry extract, and therefore proanthocyanidins, than the preparation administered with the capsules containing the "as is" extract [control capsule (F0)]. It was also surprising to find that the maximum inhibitory activity occurs at different times in the case of administration of the extract preparation (F0) (1 hour and 12 hours, P < 0.0001, Mann-Whitney test) and of the preparation (F1) (2 hours and 12 hours, P < 0.0001), which demonstrates that the longer-lasting effect of inhibition of adhesion and biofilm formation against various strains of Candida albicans, and therefore the more effective prophylaxis of recurrent urinary tract infections, is ensured by combining the American cranberry extract "as is" (i.e. not associated with phospholipids) with the composition of the present application.

Claims

1. A composition comprising: a) a Vaccinium macrocarpon extract in an amount of 20-50% by weight of the total weight of the composition; and b) at least one phospholipid in an amount of 50-80% by weight of the total weight of the composition.

2. The composition according to claim 1, wherein the Vaccinium macrocarpon extract has a proanthocyanidin content of 15-40% by weight of the total weight of the extract.

3. The composition according to claim 1 or 2, further comprising one or more technical excipients selected from water-soluble or water-insoluble diluents, lubricants and / or glidants and surfactants.

4. The composition according to claim 3, wherein the diluent is not maltodextrin, mannitol, isomalt or a combination thereof.

5. A process for the preparation of a composition comprising: a) a Vaccinium macrocarpon extract in an amount of 20-50% by weight of the total weight of the composition; and b) at least one phospholipid in an amount of 50-80% by weight of the total weight of the composition, the process comprising dissolving or dispersing the Vaccinium macrocarpon extract, the at least one phospholipid and optionally one or more technical excipients selected from water-soluble or water-insoluble diluents, lubricants and / or glidants and surfactants in an organic solvent at a temperature of 30-90°C to obtain a solution or dispersion, and then drying the solution or dispersion to obtain a solid composition.

6. The process according to claim 5, wherein the organic solvent is selected from ethanol, ethyl acetate and acetone.

7. The process according to claim 5 or 6, comprising simultaneously dissolving or dispersing the Vaccinium macrocarpon extract and the at least one phospholipid, and then optionally adding one or more excipients.

8. The process according to claim 5 or 6, comprising dissolving or dispersing the Vaccinium macrocarpon extract and the at least one phospholipid in an organic solvent to obtain a solution or dispersion, and then adding at least one phospholipid or Vaccinium macrocarpon extract to the solution or dispersion, and optionally adding one or more technical excipients.

9. The process according to any one of claims 5-8, wherein the one or more excipients are used in an amount such that their weight is 1-30% of the total weight of the composition.

10. A solid composition obtainable by the process of any one of claims 5-9.

11. Use of the composition of any one of claims 1-5 or 10 for the preparation of a pharmaceutical and / or nutritional composition for the prevention or treatment of lower urinary tract infections.

12. A solid formulation comprising the composition of any one of claims 1-5 and one or more pharmaceutically and / or nutritionally acceptable excipients mixed therewith.

13. The formulation according to claim 12, further comprising another portion of Vaccinium macrocarpon extract in free form, not mixed with phospholipids.

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