Use of Lactobacillus fermentum strains alone or in combination with Saccharomyces cerevisiae for prevention and / or treatment of vaginal candidiasis

Abstract

The present invention relates to a strain of Lactobacillus fermentum selected from the group consisting of strains preserved in CNCM at number I-5777 or number I-5778 on November 17, 2021, for use in the prevention and / or treatment of vaginal candidiasis. The present invention further relates to a strain of Lactobacillus fermentum as defined above, characterized in that it is used in combination with a strain of Saccharomyces cerevisiae preserved in CNCM at number I-3856 on October 17, 2007 for the prevention and / or treatment of vaginal candidiasis.

Description

Technical Field

[0001] The present invention relates to the field of vaginal health. In particular, the present invention relates to the use of novel Lactobacillus fermentum strains in preventing and / or treating vaginal candidiasis. The present invention also relates to the use of these novel Lactobacillus fermentum strains in combination with Saccharomyces cerevisiae in preventing and / or treating vaginal candidiasis. Background Art

[0002] Vaginal or vulvovaginal candidiasis (also called vaginal or vulvovaginal mycosis) is mainly caused by infection with pathogenic fungi, Candida albicans yeast, in the female intimate area (hence the name candidiasis). Candida albicans, which is naturally present in the vaginal flora, can proliferate abnormally and lead to the development of vaginal candidiasis. Vaginal candidiasis or vaginal mycosis causes itching and inflammation of the vulva and vagina.

[0003] Vaginal mycosis is currently considered a major public health problem and constitutes the second cause of vaginal infection after bacterial vaginosis. It is estimated that 75% of women will develop vaginal mycosis at some point in time. About 50% of them experience at least 2 to 3 episodes of vaginal mycosis in their lives. 10 to 20% of women suffer from recurrent vaginal mycosis, with an average of 4 episodes per year.

[0004] The antifungal treatment commonly used today is based on the use of compounds, mainly azole derivatives such as fluconazole, ketoconazole, econazole, miconazole or clotrimazole, usually applied in the form of creams or suppositories to be inserted into the vagina. In some cases, oral treatment is supplemented with topical treatment.

[0005] However, one of the major problems with this type of treatment is that when there are repeated episodes of vaginal mycoses, the pathogenic fungi, particularly Candida albicans, can develop resistance to existing antifungal treatments.

[0006] Another problem with the use of azole derivatives in the treatment of mycoses is irritation and a sensation of heat in the vulvar area.

[0007] In order to provide an alternative to therapies based on azole derivatives, probiotics have been described in the literature.

[0008] In 2001, the World Health Organization (WHO) defined probiotics as "live microorganisms which when administered in adequate amounts confer a health benefit beyond the effects of conventional nutritional products".

[0009] Thus, document PCT / SE2006 / 001311 describes the use of the Lactobacillus fermentum strain “Ess-1” deposited with the “Deutsche Sammlung von Mikroagencen und Zellkulturen GmbH” under the number “DSM 17851” for the prevention and / or treatment of vaginal candidiasis.

[0010] Document PCT / FR2013 / 051643 describes the use of a strain of Saccharomyces cerevisiae deposited at the CNCM under number I-3856 for the prevention and / or treatment of vaginal mycoses, in particular vaginal candidiasis.

[0011] Probiotics do not have the disadvantages of the above-mentioned chemical treatments (resistance phenomena, side effects such as irritation, burns), while providing an effectiveness at least comparable to said treatments. Probiotics therefore represent a natural alternative to chemical compounds without the side effects. SUMMARY OF THE INVENTION

[0013] Therefore, one of the objects of the present invention is to propose a novel probiotic for preventing and / or treating vaginal candidiasis.

[0014] The inventors were surprised to find that at least one fermentation Lactobacillus strain deposited on November 17, 2021 with number I-5777 or number I-5778 in CNCM (National Center for the Collection of Microorganisms, 25 Docteur Roux Street, 75724, Cedex 15, Paris, France) can achieve this purpose.

[0015] Therefore, one object of the present invention is to select a fermentation Lactobacillus strain deposited in CNCM on November 17, 2021 under number I-5777 or number I-5778.

[0016] It is important to note that regardless of whether a given strain is a bacterial strain or a yeast strain, its probiotic effect is specific to that strain and not to the genus and species in question.

[0017] Another object of the present invention is a Lactobacillus fermentum strain selected from the strains deposited in CNCM on November 17, 2021 under number I-5777 or number I-5778, for use in the prevention and / or treatment of vaginal candidiasis.

[0018] A Lactobacillus fermentum strain is a strain of bacteria or a bacterial strain. The term "bacterial strain" refers to a relatively homogeneous population of bacterial cells.

[0019] Bacterial strains are obtained from the isolation of clones, which are populations of cells obtained from single bacterial cells.

[0020] The invention also relates to bacteria obtained by culturing a strain of Lactobacillus fermentum selected from the strains deposited in CNCM on November 17, 2021 under number I-5777 or number I-5778, for use in the prevention and / or treatment of vaginal candidiasis.

[0021] In the present application, the term "bacteria" is used in the plural, but may also be used in the singular. Thus, "bacteria" means a bacterium or bacteria obtained by culturing a strain of Lactobacillus fermentum as defined above. The phrases "bacteria obtained by culturing a strain of Lactobacillus fermentum deposited at the CNCM on November 17, 2021 under the number I-5777", "bacteria obtained by culturing strain I5777" and "bacteria I-5777" are used interchangeably in the present application, and these three phrases have the same meaning as the term "bacteria" given above.

[0022] Similarly, the phrases "bacteria obtained by cultivating the Lactobacillus fermentum strain deposited in the CNCM under the number I-5778 on November 17, 2021", "bacteria obtained by cultivating strain I-5778" and "bacteria I-5778" are used interchangeably in this application, and these three phrases have the same meaning as given above.

[0023] For the purposes of this application, the terms "bacterial strain", "strain of bacteria", "Lactobacillus fermentum strain" all refer to strain I-5777 and / or strain I-5778.

[0024] For the purposes of this application, the Lactobacillus fermentum bacteria of the present invention are denoted bacteria I-5777 and / or bacteria I-5778.

[0025] As an indication, the fermentative Lactobacillus bacteria of the present invention can be obtained by culturing the strain I-5777 or the strain I-5778 overnight at pH 6.5 and 37° C. in MRS (“De Man Rogosa Sharpe”) agar medium (Biomerieux-AEB140652) without stirring.

[0026] However, fermentative lactobacillus bacteria can also be obtained by an industrial scale process comprising the following steps:

[0027] - Cultivation of bacterial strains in optimal media for biomass production,

[0028] - centrifuging the bacteria thus produced to separate them from their culture medium, to obtain a concentrated biomass in an emulsion containing 10 to 20% of dry matter, said concentrated biomass being called "centrate",

[0029] - freezing the centrifuge, preferably with liquid nitrogen,

[0030] - Possible storage of frozen centrifuges at -80°C for up to one year,

[0031] - The frozen centrifuge was lyophilized to obtain a powder containing 90 to 98% dry matter.

[0032] The powder obtained by freeze-drying corresponds to the meaning of "preferably freeze-dried dry form" in the present application.

[0033] According to one embodiment of the present invention, the bacteria as defined above for use in the prevention and / or treatment of vaginal candidiasis are in a frozen form or a dried form, preferably in a dried form, more preferably in a lyophilized dried form.

[0034] According to an advantageous embodiment of the invention, in dry form, preferably in freeze-dried form, at 11×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The bacterial strain or bacterium I-5777 is used for the prevention and / or treatment of vaginal candidiasis in a daily dose of CFU.

[0035] According to another advantageous embodiment of the invention, in dry form, preferably in freeze-dried form, at 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 A daily dose of CFU of the bacterial strain or bacterium I-5778 is used for the prevention and / or treatment of vaginal candidiasis.

[0036] The abbreviation "CFU" stands for "colony forming units".

[0037] The term "vagina" is used broadly in this application, and the terms "vagina" and "vulvovaginal" are synonymous herein.

[0038] "Vaginal candidiasis" is understood to mean a specific type of vaginal mycoses caused by fungi of the genus Candida, such as those selected from the group consisting of Candida albicans, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida glabrata and mixtures thereof.

[0039] Vaginal candidiasis is also called candidal vaginitis.

[0040] According to another embodiment, an object of the present invention is a Lactobacillus fermentum strain as defined above or a Lactobacillus fermentum bacterium as defined above, for use in the prevention and / or treatment of vaginal candidiasis caused by a fungus chosen from the group comprising Candida albicans, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida glabrata and mixtures thereof.

[0041] The Lactobacillus fermentum strain or the Lactobacillus fermentum bacterium may be used prophylactically in women susceptible to and / or sensitive to vaginal candidiasis, or may be used therapeutically, for example during an episode of infection.

[0042] The inventors have also discovered that when one of these two strains or both strains of bacteria are combined with a strain of Saccharomyces cerevisiae deposited on October 17, 2007 under the number I-3856 at the CNCM (National Collection of Microorganisms, 25 Docteur Roux Street, 15 Cedex, Paris, France, 75724), a particularly surprising synergistic effect is obtained, resulting in a particularly effective means for treating vaginal candidiasis.

[0043] In this respect, another object of the invention is a fermentative Lactobacillus strain or bacterium as defined above for the prevention and / or treatment of vaginal candidiasis, characterized in that it is used in combination with a Saccharomyces cerevisiae strain deposited with the CNCM on October 17, 2017 under number I-3856 and / or with a yeast obtained by cultivating said Saccharomyces cerevisiae strain.

[0044] Saccharomyces cerevisiae strains are yeast strains. The term "yeast strain" refers to a relatively homogenous population of yeast cells. Yeast strains are obtained from the isolation of clones, which are populations of cells obtained from a single yeast cell.

[0045] For the purposes of this application, Saccharomyces cerevisiae refers to Saccharomyces cerevisiae 1-3856.

[0046] Thus, according to one embodiment of the present invention, a Lactobacillus fermentum strain or bacteria may be combined with a Saccharomyces cerevisiae strain and / or Saccharomyces cerevisiae.

[0047] In other words, according to an advantageous embodiment of the invention, the Lactobacillus fermentum bacteria I-5777 and / or I-5778 are combined with the Saccharomyces cerevisiae bacteria I-3856.

[0048] As an indication, Saccharomyces cerevisiae can be obtained by culturing strain No. 1-3856 in a culture medium, for example as described in the reference book ("Yeast Technology", 2nd edition, 1991, G. Reed and TW Nagodawithana, published by Van Nostrand Reinhold, ISBN 0-442-31892-8).

[0049] However, Sc yeast I-3856 can also be obtained by an industrial scale process comprising the following steps:

[0050] - Cultivation of yeast strains in culture medium in several stages, first semi-anaerobic, then aerobically,

[0051] The yeast thus produced is centrifuged to separate it from its culture medium, so as to obtain a liquid yeast milk containing 12 to 25% of dry matter, or even higher amounts of dry matter, especially if the yeast milk is mixed with a permeate product.

[0052] The terms "yeast obtained by cultivating the Saccharomyces cerevisiae strain deposited in the CNCM under the number 1-3856", "yeast obtained by cultivating strain 1-3856" and "yeast Sc 1-3856" are used interchangeably in the present application, the three terms having the same meaning as given above.

[0053] The Saccharomyces cerevisiae strain deposited at the CNCM under number 1-3856 may also be referred to as "strain Sc 1-3856".

[0054] According to another advantageous embodiment of the invention, the yeast ScI-3856 used in combination with the Lactobacillus fermentum strain or bacteria is live. The live yeast is then in the form of dry yeast or fresh yeast, preferably in the form of dry yeast.

[0055] For example, in the case of yeast Sc I-3856 in combination with a Lactobacillus fermentum strain or bacteria for the prevention and / or treatment of vaginal candidiasis:

[0056] - in dry form, preferably in freeze-dried form, the fermented Lactobacillus strain or bacteria at 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 CFU daily dose, and

[0057] - Dry yeast Sc I-3856 at 1×10 7 Up to 1×10 11 CFU, preferably 1×10 9 Up to 1×10 10 CFU, even more preferably 2.5 × 10 9 Up to 5×10 9 CFU daily dose used.

[0058] Fresh yeast is characterized by a high water content. Fresh yeast is for example selected from liquid yeast or pressed yeast.

[0059] Liquid yeast, also called "liquid milky yeast", is an aqueous suspension comprising 12% to 50% yeast dry matter, more usually 12% to 25% yeast dry matter.

[0060] Compressed yeast is obtained by a / filtering liquid milky yeast, usually on a drum vacuum filter, to obtain fresh dehydrated yeast containing 26% to 37% dry matter, and b / kneading the fresh dehydrated yeast, and c / extruding.

[0061] Compressed yeast contains 26% to 37% yeast dry matter. Compressed yeast can be crushed or uncrushed.

[0062] Among dry yeasts we can cite for example active dry yeast, instant dry yeast and frozen medium moisture yeast.

[0063] One advantage of dry yeast is its long shelf life.

[0064] Active dry yeast or instant dry yeast has a yeast dry matter content of more than 90%, preferably a dry matter content of 92% to 96%.

[0065] Active dry yeast is obtained by dehydrating compressed or liquid yeast by a combined action of heat (at low temperatures) and mechanical activity, which allows the conversion of a pasty product (compressed or liquid yeast) into a dry product in pellet form. For example, active dry yeast is obtained by extrusion and drying compressed or liquid yeast in a fluidized bed.

[0066] Active dry yeast is in the form of pellets with diameters typically ranging from 0.1 μm to 2.5 mm.

[0067] Instant dry yeast is obtained by dehydrating pressed or liquid yeast, transforming the pasty product (pressed or liquid yeast) into thin dry strands by the action of a hot air gradient. Instant dry yeast must be packaged in the absence of oxygen in order to remain stable.

[0068] The frozen medium moisture yeast contains, for example, 70% to 85% yeast dry matter.

[0069] According to another advantageous embodiment of the present invention, the yeast Sc I-3856 used in combination with a Lactobacillus fermentum strain or bacteria for preventing and / or treating vaginal mycoses is in the form of active dry yeast or instant dry yeast.

[0070] According to another advantageous embodiment of the invention, the yeast Sc I-3856 used in combination with a Lactobacillus fermentum strain or bacteria for the prevention and / or treatment of vaginal mycoses is in the form of inactivated yeast (also called killed yeast).

[0071] Inactivated yeast is yeast in which metabolism has been irreversibly stopped.

[0072] Inactivated yeast can be obtained by techniques well known to those skilled in the art, such as heat treatment of the yeast, a treatment consisting of subjecting the yeast to several consecutive freeze-thaw cycles, a spraying treatment, atomization treatment or a combination of these treatments.

[0073] For example, in the case of inactivated yeast combined with Lactobacillus fermentum strains or bacteria for the prevention and / or treatment of vaginal candidiasis:

[0074] - in dry form, preferably in freeze-dried form, the fermented Lactobacillus strain or bacteria at 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 CFU daily dose, and

[0075] - Yeast Sc I-3856 in the form of inactivated yeast is used in a daily dose of 1 to 10,000 mg, preferably 100 to 1000 mg.

[0076] According to another advantageous embodiment of the present invention, the yeast ScI-3856 used in combination with the Lactobacillus fermentum strain or bacteria is used in the form of a yeast derivative selected from the cell wall of the yeast, the cell wall glucan of the yeast, the cell wall mannoprotein of the yeast or a mixture thereof.

[0077] The cell wall of yeast refers broadly to the wall and plasma membrane of yeast. Typically, the cell wall of yeast is obtained by a process comprising a yeast autolysis step followed by a step of separating the soluble fraction from the insoluble fraction, the separated insoluble fraction corresponding to the yeast wall, which may then be dried.

[0078] For example, in the case of a combination of the cell wall of yeast Sc I-3856 and a Lactobacillus fermentum strain or bacteria for the prevention and / or treatment of vaginal candidiasis:

[0079] - in dry form, preferably in freeze-dried form, the fermented Lactobacillus strain or bacteria at 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 CFU daily dose, and

[0080] - The cell wall of yeast Sc I-3856 is used in a daily dose of 0.5 to 5,000 mg, preferably 50 to 500 mg.

[0081] For example, in the case of cell wall glucans from yeast Sc I-3856 in combination with a Lactobacillus fermentum strain or bacteria for the prevention and / or treatment of vaginal candidiasis:

[0082] - in dry form, preferably freeze-dried form, of the fermented Lactobacillus strain or bacteria at 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 CFU daily dose, and

[0083] - Cell wall glucan of yeast Sc I-3856 is used in a daily dose of 0.125 to 1250 mg, preferably 12.5 to 125 mg.

[0084] As an example, in the case of a cell wall mannoprotein of yeast Sc I-3856 in combination with a Lactobacillus fermentum strain or bacteria for the prevention and / or treatment of vaginal candidiasis:

[0085] - in dry form, preferably in freeze-dried form, the fermented Lactobacillus strain or bacteria at 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 CFU daily dose, and

[0086] - The cell wall mannoprotein of yeast Sc I-3856 is used in a daily dose of 0.125 to 1250 mg, preferably 12.5 to 125 mg.

[0087] According to an advantageous embodiment of the invention, the Lactobacillus fermentum strains and / or bacteria on the one hand and the strain Sc I-3856 and / or yeast Sc I-3856 on the other hand are applied simultaneously or sequentially, preferably simultaneously.

[0088] When administered sequentially, yeast Sc I-3856 can, for example, be administered in the morning, while bacteria I-5777 and / or I-5778 are administered in the evening.

[0089] Therefore, the present invention also relates to a Lactobacillus fermentum strain and / or bacteria in combination with strain Sc I-3856 and / or yeast Sc I-3856 for simultaneous or sequential administration for the prevention and / or treatment of vaginal candidiasis.

[0090] Depending on the desired mode of administration (simultaneous or sequential), the Lactobacillus fermentum strains and / or bacteria on the one hand and the strain Sc I-3856 and / or yeast Sc I-3856 on the other hand are in the form of a single composition or in the form of separate compositions.

[0091] By a single composition is meant that the Lactobacillus fermentum strain and / or bacteria on the one hand and the strain Sc I-3856 and / or yeast Sc I-3856 on the other hand are present in a single composition.

[0092] By "isolated" composition it is meant that the Lactobacillus fermentum strain and / or bacteria is present in one composition, whereas the strain Sc I-3856 and / or yeast Sc I-3856 is present in another composition separate from the composition comprising said strain and / or bacteria.

[0093] According to a preferred embodiment, the present invention relates to the use of bacterial strains and / or bacteria I-5777 in combination with strain Sc I-3856 and / or yeast Sc I-3856 for the prevention and / or treatment of vaginal candidiasis.

[0094] Preferably, the strain and / or bacterium I-5777 on the one hand and the strain Sc I-3856 and / or yeast Sc I-3856 on the other hand are present in a single composition and are thus administered simultaneously.

[0095] According to another preferred embodiment, the present invention relates to the use of bacterial strains and / or bacteria I-5778 in combination with strain Sc I-3856 and / or yeast Sc I-3856 for the prevention and / or treatment of vaginal candidiasis.

[0096] Preferably, the bacterial strain and / or the bacterium I-5778 on the one hand and the strain Sc I-3856 and / or the yeast Sc I-3856 on the other hand are present in a single composition and are thus administered simultaneously.

[0097] Another object of the present invention relates to a composition comprising:

[0098] - a Lactobacillus fermentum strain chosen from the strains deposited at the CNCM on November 17, 2021 under number I-5777 or number I-5778, and / or a bacterium obtained by culturing said Lactobacillus fermentum strain,

[0099] - possible excipients,

[0100] It is used to prevent and / or treat vaginal candidiasis.

[0101] Another object of the present invention is a composition comprising:

[0102] - a Lactobacillus fermentum strain chosen from the strains deposited at the CNCM on November 17, 2021 under number I-5777 or number I-5778, and / or a bacterium obtained by culturing said Lactobacillus fermentum strain,

[0103] - the Saccharomyces cerevisiae strain deposited with the CNCM on October 17, 2007 under reference number I-3856 and / or yeast obtained by cultivating said Saccharomyces cerevisiae strain,

[0104] - possible excipients,

[0105] It is used to prevent and / or treat vaginal candidiasis.

[0106] The composition according to the invention for use as defined above may be a pharmaceutical composition, a food supplement or a food composition.

[0107] A food composition refers to any food, drink or confectionery. Examples of food compositions are granola bars, chewing gum, dairy products.

[0108] When the composition is a food composition, some examples of excipients are lactose, corn starch, carboxymethylcellulose, mannitol, sucrose.

[0109] "Food supplement" is understood to mean foodstuffs which are intended to supplement the normal diet and which, alone or in combination, constitute concentrated sources of nutrients or other substances having a nutritional or physiological effect.

[0110] Food supplements are marketed in dosage forms, for example in liquid or powder preparations such as capsules, lozenges, tablets, pills and other similar forms, powder sachets, liquid ampoules, bottles with droppers and other similar forms, which are intended to be taken in small dosing units.

[0111] The food compositions and food supplements are intended for oral administration.

[0112] The pharmaceutical composition of the invention is intended for oral or vaginal administration, preferably oral. The excipients used in the pharmaceutical composition are those conventionally used and suitable for the preparation of oral or vaginal forms.

[0113] Examples of pharmaceutical compositions suitable for oral administration forms include tablets, capsules, gelatin capsules, sachets, powders, emulsions, syrups or ampoules.

[0114] Examples of pharmaceutical compositions suitable for vaginal administration include suppositories, capsules, gelatin capsules, emulsions or tablets.

[0115] The daily dosage depends on the method of administration (oral or vaginal) and the type of treatment (therapeutic or prophylactic).

[0116] The composition for use as defined above is in a form suitable for oral or vaginal administration, preferably oral administration.

[0117] According to an advantageous embodiment of the invention, the composition for use as defined above does not contain any active ingredient other than the Lactobacillus fermentum strains I-5777 and / or I-5778 and / or the Lactobacillus fermentum bacteria I-5777 and / or I-5778 on the one hand, and the strain Sc I-3856 and / or the yeast Sc I-3856 on the other hand.

[0118] Therefore, the present invention also relates to a composition for use as defined above, characterized in that it comprises or consists of:

[0119] - on the one hand the Lactobacillus fermentum strains I-5777 and / or I-5778, and / or the Lactobacillus fermentum bacteria I-5777 and / or I-5778,

[0120] - strain Sc 1-3856 and / or yeast Sc 1-3856 on the other hand,

[0121] - Possible excipients.

[0122] If the composition comprises an excipient, the excipient will be selected according to the final use of the composition (pharmaceutical, food, food supplement).

[0123] According to another advantageous embodiment of the invention, the composition for use as defined above is characterized in that it also comprises an active ingredient, in particular selected from the group comprising vitamin K, vitamin B9, glutathione, S-adenosyl-L-methionine (SAM-e), chondroitin sulfate and mixtures thereof. This composition is preferably used for oral administration.

[0124] According to yet another embodiment of the invention, the composition for use as defined above is more particularly characterized in that:

[0125] - Lactobacillus fermentum strain or bacteria I-5777 and / or I-5778 in dry form, preferably lyophilized form, and in a concentration of 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The amount of CFU present, and

[0126] - Yeast Sc I-3856 was in dry form and was used at 1×10 7 Up to 1×10 11 CFU, preferably 1×10 9 Up to 1×10 10 CFU, more preferably 2.5×10 9 Up to 5×10 9 The amount of CFU present.

[0127] According to another embodiment, the composition for use as defined above is more particularly characterized in that:

[0128] - Lactobacillus fermentum strain or bacteria I-5777 and / or I-5778 in dry form, preferably lyophilized form, and in a concentration of 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The amount of CFU present, and

[0129] - Yeast Sc I-3856 is in the form of inactivated yeast and is present in an amount of 1 to 10,000 mg, preferably 100 to 1000 mg.

[0130] According to yet another embodiment, the composition for use as defined above is more particularly characterized in that:

[0131] - Lactobacillus fermentum strain or bacteria I-5777 and / or I-5778 in dry form, preferably lyophilized form, and in a concentration of 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The amount of CFU present, and

[0132] - Yeast Sc I-3856 is in the form of yeast cell walls and is present in an amount of 0.5 to 5,000 mg, preferably 50 to 500 mg.

[0133] According to another embodiment, the composition for use as defined above is more particularly characterized in that:

[0134] - Lactobacillus fermentum strain or bacteria I-5777 and / or I-5778 in dry form, preferably lyophilized form, and in a concentration of 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The amount of CFU present

[0135] - Yeast Sc I-3856 is in the form of cell wall glucan and is present in an amount of 0.125 to 1250 mg, preferably 12.5 to 125 mg.

[0136] According to another embodiment, the composition for use as defined above is more particularly characterized in that:

[0137] - Lactobacillus fermentum strain or bacteria I-5777 and / or I-5778 in dry form, preferably lyophilized form, and in a concentration of 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The amount of CFU present, and

[0138] - Yeast Sc I-3856 is in the form of a cell wall mannoprotein and is present in an amount of 0.125 to 1250 mg, preferably 12.5 to 125 mg. BRIEF DESCRIPTION OF THE DRAWINGS

[0139] Other features, details, and advantages will become apparent from reading the following detailed description and analyzing the accompanying drawings.

[0140] Figure 1

[0141] [ Figure 1] is a bar graph obtained from the first experiment, which shows the percentage of inhibition of Candida albicans hyphae formation by the lactobacilli denoted "LS1" to "LS14", Lactobacilli LS4 and LS5 corresponding to the bacterial strains of the present invention, namely strains CNCM I-5777 and CNCM I-5778, respectively.

[0142] The y-axis represents the percentage of hyphae formation normalized to Candida albicans.

[0143] The "CA" group (Candida albicans) represented the control group.

[0144] Significant differences compared with the control group are annotated with asterisks **p < 0.01.

[0145] Figure 2

[0146] [ Figure 2 ] is a bar graph obtained from the second experiment, which shows the percentage of inhibition of Candida albicans hyphae formation by the control CA and Lactobacillus LS4 (CNCM I-5777), LS5 (CNCM I-5778) and LS7, respectively.

[0147] The y-axis represents the percentage of hyphae formation normalized to Candida albicans.

[0148] Significant differences compared with the control group are annotated with asterisks ****p < 0.0001.

[0149] Figure 3

[0150] [ Figure 3 ] is a bar graph obtained from the third experiment, which shows the percentage of inhibition of Candida albicans hyphae formation by the control CA and Lactobacillus LS4 (CNCM I-5777) and LS5 (CNCM I-5778), respectively.

[0151] The y-axis represents the percentage of hyphae formation normalized to Candida albicans.

[0152] Significant differences compared with the control group are annotated with asterisks ****p < 0.0001.

[0153] Figure 4

[0154] [ Figure 4 ] is a bar graph showing the percentage of inhibition of Candida albicans hyphae formation by the control CA alone or in combination with Saccharomyces cerevisiae CNCM I-3856 ("Sc"), and by Lactobacillus (LS4, LS7 and LS11) alone (light gray bar graph) or in combination with Saccharomyces cerevisiae CNCM I-3856 (Sc) (striped bar graph).

[0155] The y-axis represents the percentage of hyphae formation normalized to Candida albicans.

[0156] "-Sc" indicates the absence of Saccharomyces cerevisiae, while "+Sc" indicates the presence of Saccharomyces cerevisiae.

[0157] Significant differences compared with the control group are annotated with asterisks ****p < 0.0001. DETAILED DESCRIPTION OF THE INVENTION

[0159] In the following Example 1, reference is made to Figures 1 to 4 . Example 1

[0160] Mycelium test

[0161] The hyphal test performed under in vitro conditions allows the evaluation of the ability of a strain (of bacteria or yeast) to block hyphal formation of Candida albicans (ie to inhibit hyphal formation).

[0162] In fact, hyphae formation is a key step in the virulence and invasiveness of C. albicans : thus, hyphal morphogenesis is an important virulence factor of C. albicans .

[0163] Hyphae inhibition / reduction represents an interesting approach to combat Candida infections.

[0164] Tested strains

[0165] Fourteen Lactobacillus strains (LS1 to LS14) were tested, alone or in combination with the Saccharomyces cerevisiae strain CNCM 1-3856.

[0166] Strains LS1 to LS14 are strains isolated from sourdough starter.

[0167] LS1: Lactiplantibacillus plantarum, identification reference BCC_002300,

[0168] LS2: Lactobacillus plantarum, identification reference BCC_002377,

[0169] LS3: Lactobacillus plantarum, identification reference BCC_003240,

[0170] LS4: Lactobacillus fermentum, identification reference BCC_002284 (strain deposited in CNCM under number I-5777 on November 17, 2021),

[0171] LS5: Lactobacillus fermentum, identification reference BCC_002321 (strain deposited in CNCM under number I-5778 on November 17, 2021),

[0172] LS6: Lactobacillus paracasei, identification reference BCC_002365,

[0173] LS7: Bacillus plantarum, identification reference BCC_002378 (strain deposited at CNCM under number I-5779 on November 17, 2021),

[0174] LS8: Lactobacillus plantarum, identification reference BCC_004104,

[0175] LS9: Lactobacillus plantarum, identification reference BCC_004132,

[0176] LS10: Lactobacillus plantarum, identification reference BCC_004299,

[0177] LS11: Lactobacillus paracasei, identification reference BCC_002274,

[0178] LS12: Lactobacillus paracasei, identification reference BCC_002301,

[0179] LS13: Lactobacillus paracasei, identification reference BCC_003027,

[0180] LS14: Lactobacillus paracasei, identification reference BCC_003258.

[0181] Test plan

[0182] Lactobacillus monotherapy

[0183] The concentrations of overnight cultures of Lactobacillus strains (LS1 to LS14) and Candida albicans strains were estimated by spectrophotometry (optical density at 600 nm). The cultures were then adjusted to 1 × 10 9 CFU / ml concentration and Candida albicans 1×10 6 The concentration of CFU / ml.

[0184] A mixture of Candida albicans and Lactobacillus was produced by adding 50 μl of a suspension of each test microorganism to 125 μl of YPD broth (yeast-peptone-dextrose broth) supplemented with fetal calf serum (FCS) to give a total volume of 500 μl.

[0185] For each condition, four technical replicates were included (results from four different wells, but from the same initial overnight culture). After incubation for 3 hours at 37°C, 2 μl of the mixture was used for microscopic evaluation to count the number of yeast cells and the number of hyphae-forming cells. At least 100 cells were counted, the ratio was calculated and normalized to the negative control (i.e., Candida albicans + FCS).

[0186] Combination of Lactobacillus and Saccharomyces cerevisiae

[0187] Supernatants from overnight cultures of Saccharomyces cerevisiae (Sc I-3856) were obtained by centrifugation (10 min, 2000 g) and filtration (0.20 μm cellulose acetate filter).

[0188] The concentration of overnight cultures of Lactobacillus and Candida albicans strains was estimated by spectrophotometry (optical density at 600 nm). The cultures were then adjusted to 1 × 10 9 The concentration of CFU / m and Candida albicans 1×10 6 The concentration of CFU / ml.

[0189] A mixture of Candida albicans and Lactobacillus was produced by adding 50 μl of a suspension of each test microorganism to 125 μl of YPD broth supplemented with fetal calf serum (FCS) to give a total volume of 500 μl.

[0190] The rest of the regimen was the same as that for Lactobacillus alone described above.

[0191] result

[0192] The results obtained are described below and in Figures 1 to 4 It is noted that in each of these figures, the inhibition of hyphae formation by the control CA (Candida albicans) was 0 percent (0%) (which means 100% of hyphae formation by C. albicans).

[0193] Lactobacillus alone ( Figures 1 to 3 )

[0194] from Figure 1 It can be clearly seen that, among the 14 lactobacilli tested (LS1 to LS14), only Lactobacillus fermentum strains LS4 (CNCM I-5777) and LS5 (CNCM I-5778) provided strong inhibition of hyphae formation of Candida albicans.

[0195] Indeed, for strains LS4 and LS5, about 72% inhibition of hyphae formation of C. albicans (ie hyphae formation about 28%) (LS4) and about 78% inhibition of hyphae formation of C. albicans (ie hyphae formation about 22%) (LS5) were observed, respectively.

[0196] exist Figure 2 In the present study, strains LS4 (CNCM I-5777) and LS5 (CNCM I-5778) were compared with strain LS7 (CNCM I-5779), since this strain represented a large number of strains tested.

[0197] from Figure 2 It is also clear that strains LS4 (CNCM I-5777) and LS5 (CNCM I5778) provide a strong inhibition of Candida albicans hyphae formation compared to strain LS7 (CNCM I-5779). In fact, for strains LS4 and LS5, we observed about 74% inhibition (LS4) and about 75% inhibition (LS5), respectively, while for strain LS7 (CNCM I-5779), only 20% inhibition.

[0198] exist Figure 3 In the comparison, only strains LS4 (CNCM I-5777) and LS5 (CNCM I-5778) were compared. Once again, the results obtained are promising, since these strains lead to a strong inhibition of Candida albicans hyphae formation, namely about 81% (LS4) and 93% (LS5) inhibition of Candida albicans hyphae formation, respectively.

[0199] Combination of Lactobacillus and Saccharomyces cerevisiae ( Figure 4 )

[0200] from Figure 4 It can be clearly seen that the combination of LS4 (CNCM I-5777) with S. cerevisiae CNCM I-3856 leads to an unexpected synergistic effect, which is not the case in the combination of strains LS7 (CNCM I-5779) and LS11 with Sc I-3856, respectively.

[0201] Indeed, under the experimental conditions in question, the use of the Saccharomyces cerevisiae strain CNCM I-3856 (see "CA + Sc") alone (ie not in combination with Lactobacillus) results in an inhibition of Candida albicans hyphae formation of about 9%. The use of the strain LS4 alone (see "LS4-Sc") results in an inhibition of Candida albicans hyphae formation of about 52%. However, when the strains LS4 and Sc CNCM I-3856 are combined, the inhibition of Candida albicans hyphae formation is almost complete, since it is about 97%.

[0202] The invention is not limited to the embodiments described above, which are given as examples only, but covers all variants that are conceivable to a person skilled in the art within the scope of the claimed protection.

[0203] References to biological materials deposited in repositories

[0204] In this application, reference is made to the following biological materials and derived variants or mutant strains:

[0205] - Identification reference "BCC_002284" and accession number "I-5777", deposited on November 17, 2021 by Lesaffre SA, 41 rue Etienne Marcel 75009 Paris, at the National Collection of Microorganisms (CNCM), 25 rue Docteur Roux, 15 Cedex, 75724 Paris, France;

[0206] - Identification reference "BCC_002321" and accession number "I-5778", deposited on November 17, 2021 by Lesaffre SA, 41 rue Etienne Marcel 75009 Paris, at the National Collection of Microorganisms (CNCM), 25 rue Docteur Roux, 15 Cedex, 75724 Paris, France;

[0207] - Identification reference "BCC_002378" and accession number "I-5779", deposited on November 17, 2021 by Lesaffre SA, 41 rue Etienne Marcel 75009 Paris, at the National Collection of Microorganisms (CNCM), 25 rue Docteur Roux, 15 Cedex, 75724 Paris, France;

[0208] - Identified with reference "SCPro-1" and accession number "I-3856", deposited on October 17, 2007 by Lesaffre SA, 41 rue Etienne Marcel 75009 Paris, at the National Collection of Microorganisms (CNCM), 25 rue Docteur Roux, 15 Cedex, 75724 Paris, France.

Claims

1. A fermented Lactobacillus strain selected from the strains deposited in CNCM on November 17, 2021 under number I-5777 or number I-5778, for use in preventing and / or treating vaginal candidiasis.

2. Bacteria obtained by culturing a Lactobacillus fermentum strain selected from the strains deposited in CNCM on November 17, 2021 under number I-5777 or number I-5778, for use in the prevention and / or treatment of vaginal candidiasis.

3. The bacteria for use according to claim 2, characterized in that It is present in frozen form or in dried form, preferably in dried form, more preferably in lyophilized dried form.

4. The strain for use according to claim 1 or the bacterium for use according to claim 2 or 3, wherein the vaginal candidiasis is caused by a fungus selected from the group consisting of Candida albicans, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida glabrata and mixtures thereof.

5. The strain or bacterium for use according to any one of claims 1 to 4, wherein it is used in combination with the Saccharomyces cerevisiae strain deposited at the CNCM on October 17, 2007 under the number I-3856 and / or with a yeast obtained by cultivating said Saccharomyces cerevisiae strain.

6. The strain or bacterium for use according to claim 5, wherein the yeast obtained by culturing the Saccharomyces cerevisiae strain is in the form of dry yeast or fresh yeast, preferably in the form of dry yeast.

7. The strain or bacterium for use according to claim 6, wherein the dry yeast is in the form of active dry yeast or instant dry yeast.

8. The strain or bacterium for use according to claim 5, wherein the yeast is in the form of an inactivated yeast.

9. The strain or bacterium for use according to claim 5, wherein the yeast is in the form of a yeast derivative selected from the group consisting of the cell wall of the yeast, the cell wall glucan of the yeast, the cell wall mannoprotein of the yeast or a mixture thereof.

10. A composition comprising: - a Lactobacillus fermentum strain chosen from the strains deposited at the CNCM on November 17, 2021 under number I-5777 or number I-5778, and / or a bacterium obtained by culturing said Lactobacillus fermentum strain, - the Saccharomyces cerevisiae strain deposited with the CNCM on October 17, 2007 under reference number I-3856 and / or yeast obtained by cultivating said Saccharomyces cerevisiae strain, - possible excipients, It is used to prevent and / or treat vaginal candidiasis.

11. The composition for use according to claim 10, wherein the composition is a pharmaceutical composition, a food composition or a food supplement.

12. The composition for use according to claim 10 or 11, wherein the composition is in a form suitable for oral or vaginal administration, preferably oral administration.

13. The composition for use according to any one of claims 10 to 12, further comprising an active ingredient selected from the group comprising vitamin K, vitamin B9, glutathione, S-adenosyl-L-methionine (SAM-e), chondroitin sulfate and mixtures thereof.

14. A composition for use according to any one of claims 10 to 13, wherein: - Lactobacillus fermentum strain or bacteria I-5777 and / or I-5778 in dry form, preferably lyophilized form, and in a concentration of 1×10 7 Up to 1×10 10 CFU, preferably 2×10 8 Up to 2×10 9 The amount of CFU is present, and - Yeast Sc I-3856 was in dry form and was used at 1×10 7 Up to 1×10 11 CFU, preferably 1×10 9 Up to 1×10 10 CFU, even more preferably 2.5 × 10 9 Up to 5×10 9 The amount of CFU present.

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