Compositions for promoting growth of lactobacillus crispatus

Abstract

The present invention relates to methods and compositions for promoting and increasing growth of Lactobacillus crispatus.

Description

[0001] COMPOSITIONS FOR PROMOTING GROWTH OF LACTOBACILLUS CRISPATUS

[0002] Technical field

[0003] The present invention relates to methods and compositions for promoting and increasing growth of Lactobacillus crispatus.

[0004] Background

[0005] Lactobacillus crispatus and Gardnerella vaginalis are among the most prominent vaginal bacteria associated with health and disease of the vagina. While Lactobacillus species are associated with a healthy microflora due to their production of hydrogen peroxide and lactic acid which lowers the vaginal pH and discouraging the growth of anaerobic bacteria, G. vaginalis is one of the species associated with bacterial vaginosis (BV) and considered a key component of the anaerobic diversity responsible for symptoms found in BV patients. G. vaginalis are known for their production of a biofilm, clusters of bacteria attached to the vaginal epithelia. Bacteria in a biofilm are protected from the immune system and antibiotics. A BV infection is proceeded by what is referred to as “a shift in vaginal flora”, whereby the Lactobacillus species are lost, and this shift paves way for the anaerobic pathogenic bacteria, e.g. G. vaginalis (Kalia et al. 2020, Falconi-McCahill 2019).

[0006] A disrupted reproductive tract microbiome has been associated with unfavourable outcomes of pregnancy and reproductive complications. Ascending bacterial infection from the vagina through the cervix into the uterine cavity is considered to be a major cause of preterm birth (Goldenberg et al, Lancet, 2008, 37, 9606, 75-849; MacIntyre et al, Sci Rep, 2015, 5, 8988). Many of these cases also exhibit chorioamnionitis, constituting a major threat also to the mother, before the pathogen is identified and effective antibiotic treatment is commenced. The link between vaginal microbial community structure and ascending infection is also highlighted in late miscarriage, in recurrent spontaneous abortion, in chronic cervicitis / endometritis leading to implantation problems and female infertility. A healthy vaginal microbiome during pregnancy should be dominated by only one or a few Lactobacillus species (MacIntyre et al, Sci Rep, 2015, 5, 8988, Romero et al, Microbiome, 2014, 2, 1 ,4), something that is an acknowledged problem with a great medical need among pregnant women. Summary

[0007] It has previously been shown that glucono-delta-lactone (GDL) inhibits bacterial growth, for example the growth of pathogenic bacteria of the vaginal microbiome such as Gardnerella vaginalis. In particular, it has previously been shown that a lower minimal inhibitory concentration (MIC) of GDL is required to inhibit the growth of G. vaginalis as compared to the MIC required to inhibit the growth of L. crispatus (Example 13, Table 9 of WO 2019 / 0668862) and thus that GDL is more efficient against G. vaginalis than against L. crispatus. Thus, administration of GDL at a suitable concentration may enable simultaneous decrease of levels of G. vaginalis and maintenance of levels of L. crispatus.

[0008] The inventors of the present invention have however surprisingly found that administration of GDL in fact increases the growth of Lactobacillus crispatus. In other words, GDL enables and promotes growth of L. crispatus rather than inhibits growth of L. crispatus at relevant concentrations. Hence, the present invention provides methods and compositions for increasing (i.e. promoting) growth of L. crispatus comprising administering said composition to a subject in need thereof.

[0009] In one aspect, the present invention relates to a method of increasing growth and / or reproduction of Lactobacillus crispatus in a subject, the method comprising administering to the subject a composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof.

[0010] In one aspect, the present invention relates to use of a composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof, for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

[0011] In one aspect, the present invention relates to a composition comprising glucono-delta- lactone (GDL), or a pharmaceutically acceptable salt thereof, for use for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

[0012] In one aspect, the present invention relates to an in vitro method of increasing growth and / or reproduction of Lactobacillus crispatus, the method comprising incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising glucono-delta-lactone (GDL),

[0013] In one aspect, the present invention relates to an in vitro method of growing Lactobacillus crispatus, the method comprising incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising glucono-delta- lactone (GDL),

[0014] Description of Drawings

[0015] Figure 1. Growth of L. crispatus for up to 24h in the absence (empty circles) or presence of 0.5% (black circles), 2.5% (black squares), 5% (black triangles) of glucono delta lactone and sodium gluconate as measured by OD600. The experiment was performed without oil overlay, i.e. in aerobic conditions.

[0016] Figure 2. Growth of G. vaginalis for up to 24h in the absence (empty circles) or presence of 0.5% (black circles), 2.5% (black squares), 5% (black triangles) of glucono delta lactone and sodium gluconate as measured by OD600. The experiment was performed with oil overlay, i.e. in anaerobic conditions.

[0017] Figure 3. Average change per person in abundance of total Lactobacilli between day 0 (before treatment) and day 7 (after treatment), as measured with qPCR of vaginal samples from patients treated with Formulation A tablet (black bar, n=103) and a placebo tablet (empty bar, n=25).

[0018] Figure 4. Changes in abundance of total Lactobacilli per person between day 0 (before treatment) and day 7 (after treatment), as measured with qPCR of vaginal samples from patients treated with Formulation A tablet (n=103) divided into the percentage of patients having an increase at Day 7 compared to Day 0 in abundance (black bar), decrease (white bar) and no change (grey bar).

[0019] Figure 5. Changes in abundance of Lactobacillus crispatus (L. crispatus', black bar), and Gardnerella vaginalis (G. vaginalis’, white bar) respectively per person between day 0 (before treatment) and day 7 (after treatment), as measured with qPCR of vaginal samples from patients treated with Formulation A tablet (n=103).

[0020] Figure 6. Classification of the vaginal samples bacterial composition, as measured with qPCR, Day 0 (before treatment) and Day 7 (after treatment) into different community state types (CST) based on the Vaginal Community State Type Nearest Centroid Classifier (VALENCIA, France et al. 2020). Most of the samples falls in category IV-B at day 0 (before treatment), which is the community state type (CST) associated with a low relative abundance of any Lactobacillus species and high to moderate abundance of G. vaginalis and A. vaginae. For patients treated with Formulation A (n=103), there is a shift in CST category at day 7 (after treatment) as compared to Day 0 (before treatment) to a lower CST category, mainly to CST III, which denotes a vaginal microbiome community with increased levels of Lactobacilli and reduction of vaginal dysbiosis. For placebo (n=25), this shift is not seen. Detailed description

[0021] Definitions

[0022] The terms “dysbiosis” or dysbiotic” as used herein refers to a disrupted microbiome resulting in an imbalance in the microbiota and a shift in the distribution of microbial species of the microbiome as compared to a non-disrupted, balanced and healthy microbiome. Preferably, the term as used herein refers to a dysbiosis in the urogenital, such as in the vaginal, microbiome, wherein the normally dominating species, such as Lactobacillus in the vaginal microbiome, have become underrepresented and the normally non-dominating species, such as Gardnerella vaginalis, Escherichia coli, Mobiluncus spp., Staphylococcus aureus etc. in the vaginal microbiome, have increased or taken over. Dysbiosis of the vaginal microbiome can result in various conditions, for example bacterial vaginosis. Whether or not a vaginal microbiome is dysbiotic can be determined by methods known in the art, for example as described by France et al., 2020.

[0023] The term “increasing growth and / or reproduction” as used herein refers to the process of increasing, promoting or enabling growth of a microorganism. In other words, the term refers to the process of increasing, promoting or enabling the division of microbial cells, such as L. crispatus cells, so that the total number (i.e. the level or amount) of microbial cells increases over time. The increase in growth or reproduction of microbial cells can be determined using various methods that are well known in the art. For example, cell growth or reproduction can be measured by measuring the optical density (OD) (i.e. the absorbance at 600 nm) of a cell culture over time. If the OD remains constant or decreases over time, no growth or reproduction has occurred. If the OD increases over time, growth or reproduction has occurred. The growth or reproduction can also be measured by quantitative polymerase chain reaction (qPCR) (also over time, e.g. at two time point), which can provide the absolute or relative amount of a bacterial species, such as L. crispatus, in a microbiome, such as in the vaginal microbiome. If the absolute or relative amount of a bacterial species increased over time, growth or reproduction of the bacteria has occurred.

[0024] The growth and / or reproduction of a specific microorganism may for example have increased after administration of a composition as described herein, wherein said increase is relative to the growth and / or reproduction of the same microorganism grown under the same conditions but in the absence of said composition.

[0025] Compositions and methods for increasing growth of Lactobacillus crispatus

[0026] The present invention relates to a method of increasing growth and / or reproduction of Lactobacillus crispatus in a subject, the method comprising administering to the subject a composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof.

[0027] The present invention further relates to use of a composition comprising glucono-delta- lactone (GDL), or a pharmaceutically acceptable salt thereof, for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

[0028] The present invention further relates to a composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof, for use for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

[0029] The expressions “the composition comprising the compound” and “the composition comprising GDL” are used interchangeably herein. In one embodiment, the composition further comprises sodium gluconate (NaG).

[0030] In one embodiment, the composition is administered to the urogenital system of the subject. Hence, in one embodiment, the composition for use if administered to the urogenital system of the subject.

[0031] As stated above under “Definitions”, the term “increasing growth and / or reproduction” as used herein refers to increasing the amount, such as the level, of the relevant microorganism. The increased growth and / or reproduction may be absolute and / or relative. Hence, the increase may for example be an increase in absolute abundance of the microorganism and / or an increase in relative abundance, such as abundance in relation to other microorganisms in e.g. the microbiome. Preferably, the increased is an increase in absolute abundance, The increase in growth and / or reproduction may be measured by using methods known in the art, such as for example by qPCR, by measuring in vitro growth, and or by measuring colony forming units (CFU) over time.

[0032] In a preferred embodiment, the increase in growth and / or reproduction of a specific microorganism, such as L. crispatus, takes place after administration of a composition as described herein, such as a composition comprising GDL, wherein said increase is relative to the growth and / or reproduction of the same microorganism in the same conditions but in the absence of said composition.

[0033] Hence, in one embodiment, the growth and / or reproduction of L. crispatus is increased as compared to the growth and / or reproduction of L. crispatus grown in the absence of the composition disclosed herein, but otherwise grown and / or cultivated under the same conditions.

[0034] In a preferred embodiment, the growth and / or reproduction of L. crispatus is increased in a subject after administration of the composition disclosed herein, wherein the increase is as compared to the growth and / or reproduction of L. crispatus in said subject but prior to being administered the composition disclosed herein. In other words, the level, such as the amount, of L. crispatus is increased in a subject after administration of the composition disclosed herein, wherein the increase is as compared to the level, such as the amount, of L. crispatus in said subject but prior to being administered the composition disclosed herein. In yet another embodiment, the growth and / or reproduction of L. crispatus is increased in a subject after administration of the composition disclosed herein, wherein the increase is as compared to, i.e. relative to, the growth and / or reproduction of L. crispatus in another subject being administered a placebo composition. In other words, the level, such as the amount, of L. crispatus is increased in a first subject after administration of the composition disclosed herein, wherein the increase is as compared to the level, such as the amount, of L. crispatus in another subject that is not being administered the composition disclosed herein. Preferably, both subjects suffer from the same condition, such as wherein both subjects suffer from bacterial vaginosis.

[0035] In one embodiment, the level, such as the amount, of L. crispatus is increased at least 1 .5-fold, such as at least 2-fold, such as at least 5-fold, such as at least 10-fold, such as at least 20-fold, such as at least 50-fold, such as at least 100-fold, such as at least 500-fold, such as at least 1000-fold, such as at least 5000-fold, such as at least 10,000-fold.

[0036] In one embodiment, the level of L. crispatus is increased between 1.5-fold and 10,000- fold, such as between 1.5-fold and 10-fold, such as between 1000-fold and 10,000-fold, such as between 4000-fold and 6000-fold, such as between 3000-fold and 7000-fold, such as between 10-fold and 500-fold, such as between 100-fold and 300-fold.

[0037] In one embodiment, the increase is as compared to the level of L. crispatus prior to administration of the composition comprising the compound, such as wherein the increase is an increase in the level of L. crispatus in a subject after administration of the composition disclosed herein as compared to the level of L. crispatus in the same subject prior to administration of the composition disclosed herein.

[0038] In one embodiment, the increase is an increase as in the level of L. crispatus in a subject after administration of the composition disclosed herein as compared to the level of L. crispatus in another subject that has not been administered the composition disclosed herein.

[0039] In one embodiment, the level of L crispatus is increased after at least 1 day of administration, such as at least 3 days of administration, such as after at least 6 days of administration, such as after at least 7 days of administration of the composition disclosed herein.

[0040] In one embodiment, the level of L crispatus remains increased for at least 14 days after administration, such as for at least 35 days after administration, such as for at least 4 months after administration of the composition disclosed herein.

[0041] In one embodiment, the level of L. crispatus is measured as the absolute abundance, such as for example the absolute abundance of L. crispatus in the urogenital microbiome, such as the vaginal microbiome, of a subject.

[0042] In one embodiment, the level of L. crispatus is measured as the mean relative abundance, such as for example the mean relative abundance of urogenital microbiome, such as the vaginal microbiome, of a subject.

[0043] In one embodiment, the increase in relative abundance of L. crispatus after administration of the composition disclosed herein is at least 50-fold, such as at least 75-fold, such as at least 100-fold, such as at least 200-fold, such as at least 300-fold, such as at least 400-fold, such as at least 500-fold.

[0044] Methods for measuring levels L. crispatus in a subject are well known in the art. Such methods include, for example, qPCR of and / or measuring the amount of colony forming units (CFUs) in a sample from the subject.

[0045] In one embodiment, the mean relative abundance of L. crispatus is at least 10%, such as at least 20%, such as at least 30%, such as at least 40%, such as at least 50%, such as at least 60% after the final day of administration.

[0046] In yet another aspect, the present invention relates to an in vitro method of increasing growth and / or reproduction of Lactobacillus crispatus, the method comprising incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising glucono-delta-lactone (GDL),

[0047] Thus, in one aspect, the present invention relates to an in vitro method of growing Lactobacillus crispatus, the method comprising incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising glucono-delta- lactone (GDL),

[0048] In one embodiment, the concentration of GDL, such as the concentration of GDL in the composition or in the growth medium, is at least 0.75 wt%, such as at least 1 wt%, such as at least 1.5 wt%, such as at least 2 wt%, such as at least 2.5 wt%, such as at least 5 wt%, such as at least 10 wt%, such as at least 15 wt%, such as at least 20 wt%, such as at least 25 wt%.

[0049] In one embodiment, the concentration of GDL, such as the concentration of GDL in the composition or in the growth medium, is at most 30 wt%, such as at most 35 wt%, such as at most 20 wt%, such as at most 15 wt%, such as at most 10 wt%, such as at most 7.5 wt%, such as at most 5 wt%.

[0050] In one embodiment, the concentration of GDL, such as the concentration of GDL in the composition or in the growth medium, is between 1 wt% and 30 wt%, such as between 5 wt% and 20 wt%, such as between 10 wt% and 20 wt%, such as between 1 wt% and 10 wt%, such as between 1 wt% and 7.5 wt%.

[0051] In one embodiment, the increase in growth and / or reproduction is relative to the growth and / or reproduction of L. crispatus prior to incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising GDL. In one embodiment, the increase in growth and / or reproduction is relative to the growth and / or reproduction of L. crispatus in a similar growth medium not comprising or consisting of a composition comprising GDL.

[0052] Administration of the composition

[0053] The composition may be administered in any way suitable. In one embodiment, the composition is administered vaginally. Hence, in one embodiment the method comprises vaginal administration of the composition.

[0054] In one embodiment, the composition is administered at least once daily, such as at least twice daily, such as at least three times daily. Hence, in one embodiment, the method comprises administering the composition is administered at least once daily, such as at least twice daily, such as at least three times daily.

[0055] In one embodiment, the composition is administered no more than every second day, such as no more than every third day, such as no more than once a week. Hence, in one embodiment, the method comprises administering the composition no more than every second day, such as no more than every third day, such as no more than once a week.

[0056] In one embodiment, the composition is administered for no more than six days. Hence, in one embodiment, the method comprises administering the composition for no more than six days.

[0057] In one embodiment, the composition is administered for at least one week, such as for at least two weeks, such as for at least three weeks, such as for at least four weeks. Hence, in one embodiment, the method comprises administering the composition for at least one week, such as for at least two weeks, such as for at least three weeks, such as for at least four weeks.

[0058] In one embodiment, the composition adapted for administration for: a. at least once daily, such as at least twice daily, such as at least three times daily; and / or b. no more than every second day, such as no more than every third day, such as no more than once a week; and / or c. no more than six days; and / or d. at least one week, such as for at least two weeks, such as for at least three weeks, such as for at least four weeks; and / or e. at least once daily during at least a week.

[0059] Hence, in one embodiment, the method comprises administering the composition according to a., b., c., d., and / or e. above.

[0060] In one embodiment, the composition is administered vaginally once daily for at least six days.

[0061] In one embodiment, GDL is released for a prolonged time upon administration of the composition. In one embodiment, GDL is released for at least 6 h in vivo, such as for at least 12 h, such as for at least 18 h, such as for at least 24 h, such as for at least 36 h, such as for at least 42 h in vivo. In one embodiment, GDL is released for at least 24 h, such as for at least 36 h in vivo. In one embodiment, GDL is released for no more than 72 h, such as for no more than 48 h in vivo.

[0062] Formulation of the composition

[0063] In one embodiment, the composition further comprises gluconic acid, or a pharmaceutically acceptable salt thereof.

[0064] In water solution, gluconic acid is in equilibrium with glucono-y-lactone and GDL. Thus, in one embodiment, GDL (structure to the left in the figure below) is in equilibrium with gluconic acid (structure in the middle in the figure below) and glucono-y-lactone (structure to the right in the figure below) in aqueous solution. In one embodiment, the composition further comprises sodium gluconate (NaG).

[0065] In one embodiment, the composition further comprises glucono-gamma-lactone, or a pharmaceutically acceptable salt thereof.

[0066] In one embodiment, the composition comprises at least 5 wt %, such as at least 10 wt %, such as at least 15 wt %, such as at least 20 wt %, such as at least 25 wt %, such as at least 30 wt %, such as at least 40 wt %, such as at least 50 wt %, such as at least 60 wt % of GDL.

[0067] In one embodiment, the composition comprises no more than 99 wt %, such as no more than 95 wt %, such as no more than 90 wt %, such as no more than 85 wt %, such as no more than 80 wt %, such as no more than 75 wt % of GDL.

[0068] In one embodiment, the composition comprises 5 to 99 wt %, such as in 10 to 95 wt %, such as in 15 to 95 wt %, such as in 20 to 90 wt %, such as in 40 to 95 wt %, such as in 40 to 95 wt %, such as in 50 to 95 wt % of GDL.

[0069] In one embodiment, the composition comprises no more than 10 wt % water, preferably no more than 5 wt % water.

[0070] In one embodiment, the composition comprises: a. Glucono-delta-lactone (GDL); and b. A gluconate salt, such as sodium gluconate (NaG).

[0071] In one embodiment, the composition comprises: a. Glucono-delta-lactone (GDL); and b. Hypromellose (HPMC).

[0072] In one embodiment, the composition comprises: a. Glucono-delta-lactone (GDL); b. A gluconate salt; and c. Hypromellose (HPMC). In one embodiment, the gluconate salt is selected from the group consisting of sodium gluconate, potassium gluconate, calcium gluconate, magnesium gluconate, iron(ll)gluconate, copper(ll)gluconate, and zinc gluconate.

[0073] In one embodiment, the composition comprises: a. Glucono-delta-lactone (GDL); b. Sodium gluconate (NaG); and c. Hypromellose (HPMC).

[0074] In one embodiment, the composition is formulated as a tampon, vagitorium, vaginal aerosol, vaginal cup, vaginal gel, vaginal insert, vaginal patch, vaginal ring, vaginal sponge, vaginal suppository, vaginal cream, vaginal emulsion, vaginal foam, vaginal lotion, vaginal ointment, vaginal powder, vaginal shampoo, vaginal solution, vaginal spray, vaginal suspension, vaginal tablet, vaginal rod, vaginal disc, vaginal device, and any combination thereof, or wherein the pharmaceutical formulation is present on a sanitary article, such as a tampon, a sanitary napkin, an incontinence pad or diaper, or a panty liner.

[0075] In one embodiment, the composition is formulated as a vagitorium, vaginal insert, vaginal ring, vaginal suppository, vaginal tablet, vaginal rod, or vaginal disc, preferably wherein the composition is formulated as a vaginal tablet.

[0076] In one embodiment, the composition is formulated as a vaginal pessary. In one embodiment, the composition is in the form of a pharmaceutical dosage form. In one embodiment, the composition is a pharmaceutical formulation. In a preferred embodiment, the composition is a tablet.

[0077] In one embodiment, the mass of the tablet is at least 0.5 g, such as at least 0.7 g, such as at least 0.85 g, such as at least 1.0 g, such as at least 1.25 g, such as at least 1.4 g.

[0078] In one embodiment, the mass of the tablet is no more than 2.0 g, such as no more than 1.75 g, such as no more than 1.5 g, such as no more than 1.0 g, such as no more than 0.9 g. In one embodiment, the mass of the tablet is in the range of 0.5 to 2.0 g, such as in the range of 0.7 to 1.75 g, such as in the range of 1.4 to 1.6 g, such as about 1.5 g. In one embodiment, the mass of the tablet is in the range of 0.5 to 2.0 g, such as in the range of 0.5 to 1.25 g, such as in the range of 0.7 to 1.0 g, such as about 0.85 g.

[0079] In one embodiment, the tablet is bullet shaped.

[0080] In one embodiment, the tablet has a width of at least 5 mm, such as at least 7 mm, such as at least 9 mm, such as at least 10 mm. In one embodiment, the tablet has a width of no more than 15 mm, such as no more than 12 mm, such as no more than 11 mm, such as no more than 10 mm. In one embodiment, the tablet has a width in the range of 8 to 12 mm, such as in the range of 8.5 to 11 mm, such as 10 to 10.5 mm. In one embodiment, the tablet has a width in the range of 7.0 to 12 mm, such as in the range of 8.0 to 10 mm, such as 8.5 to 9.5 mm.

[0081] In one embodiment, the tablet has a length of at least 10 mm, such as at least 15 mm, such as at least 20 mm. In one embodiment, the tablet has a length of no more than 30 mm, such as no more than 25 mm, such as no more than 20 mm. In one embodiment, the tablet has a length in the range of 10 to 30 mm, such as in the range of 15 to 20 mm, such as in the range of 16 to 18 mm. In one embodiment, the tablet has a length in the range of 10 to 30 mm, such as in the range of 22 to 27 mm, such as in the range of 24 to 25 mm. In one embodiment, the tablet has a length of 24.5 mm, a width of 10.3 mm and a height of 6.5 mm.

[0082] In one embodiment, the amount of GDL is 50 to 500 mg, such as 100 to 400 mg, such as 250 to 350 mg, such as about 300 mg.

[0083] In one embodiment, the composition comprises at least 5 wt% GDL, such as at least 10 wt%, such as at least 15 wt%, such as at least 20 wt% GDL. In one embodiment, the composition comprises no more than 50 wt% GDL, such as no more than 40 wt%, such as no more than 35 wt%, such as no more than 30 wt%, such as no more than 25 wt%, such as no more than 20 wt% GDL. In one embodiment, the composition comprises in the range of 10 to 30 wt% GDL, such as 15 to 25 wt% GDL, such as about 20 wt% GDL.

[0084] In one embodiment, the composition comprises a gluconate salt in at least a concentration that corresponds to an equimolar concentration of 10 wt% NaG, such as of at least 15 wt% NaG, such as of at least 20 wt% NaG. In one embodiment, the composition comprises at least 10 wt% NaG, such as at least 15 wt% NaG, such as at least 20 wt% NaG. In one embodiment, the composition comprises no more than 30 wt% NaG, such as no more than 25 wt%, such as no more than 20 wt% NaG. In one embodiment, the composition comprises in the range of 10 to 30 wt% NaG, such as 15 to 25 wt%, such as 20 to 25 wt%, such as 24 to 25 wt% NaG.

[0085] In one embodiment, the molar ratio of GDL and gluconate salt is 2:1 to 1:2, such as 3:2 to 2:3, such as about 1:1. In one embodiment, the molar ratio of GDL and NaG is 2:1 to 1 :2, such as 3:2 to 2:3, such as about 1:1.

[0086] In one embodiment, the HPMC is selected from the group consisting of HPMC substitution types 2208, 2910, 1828 and 2906. In one embodiment, the HPMC is of substitution type 2208 or 2910.

[0087] In one embodiment, the HPMC has a viscosity of at least 10 mPas, such as at least 15 mPas, such as at least 20 mPas, such as at least 25 mPas, such as at least 30 mPas, such as at least 35 mPas, such as at least 40 mPas, such as at least 45 mPas, such as at least 50 mPas, such as at least 55 mPas, such as at least 60 mPas, such as at least 65 mPas, such as at least 70 mPas, such as at least 75 mPas.

[0088] In one embodiment, the HPMC has a viscosity of no more than 400 000 mPas, such as no more than 300 000 mPas, such as no more than 200 000 mPas, such as no more than 150 000 mPas, such as no more than 100 000 mPas, such as no more than 50 000 mPas, such as no more than 10 000 mPas, such as no more than 1 000 mPas, such as no more than 500 mPas, such as no more than 250 mPas.

[0089] In one embodiment, the HPMC has a viscosity in the range of 50 to 200 mPas, such as about 100 mPas. In another embodiment, the HPMC has a viscosity in the range or 50 000 mPas to 200 000 mPas, such as about 100 000 mPas.

[0090] In one embodiment, the composition comprises at least 5 wt% HPMC, such as at least 10 wt%, such as at least 10 wt%, such as at least 15 wt%, such as at least 20 wt%, such as at least 25 wt%, such as at least 30 wt%, such as at least 35 wt%, such as at least 40 wt%, such as at least 45 wt%, such as at least 50 wt% HPMC. In one embodiment, the composition comprises no more than 60 wt% HPMC, such as no more than 55 wt%, such as no more than 50 wt%, such as no more than 45 wt%, such as no more than 45 wt%, such as no more than 40 wt%, such as no more than 35 wt%, such as no more than 30 wt%, such as no more than 25 wt%, such as no more than 20 wt%, such as no more than 15 wt%, such as no more than 10 wt% HPMC. In one embodiment, the composition comprises in the range of 5 to 60 wt% HPMC, such as 15 to 40 wt%, such as 20 to 35 wt% HPMC.

[0091] In one embodiment, the composition further comprises silica. In one embodiment, the silica is selected from the group consisting of colloidal anhydrous silica, colloidal silicone dioxide, colloidal hydrated silica and hydrophobic colloidal silica. In one embodiment, the composition comprises in the range of 0.1 to 5 wt% silica, such as 0.1 to 2 wt%, such as about 0.5 wt% silica.

[0092] In one embodiment, the composition further comprises a lubricant. In one embodiment, the lubricant is selected from the group consisting of magnesium stearate, calcium stearate, stearic acid, sodium stearylfumarate, glyceryl behenate, glyceryl monooloeate, glyceryl monostearate, glyceryl palmitostearate, sucrose palmitate and sucrose stearate. In one embodiment, the lubricant is magnesium stearate.

[0093] In one embodiment, the composition comprises in the range of 0.1 to 5 wt% lubricant, such as 0.1 to 2 wt%, such as 0.70 to 1.3 wt%, such as about 1 wt% lubricant.

[0094] In one embodiment, the composition further comprises starch. In one embodiment, the starch is pregelatinised starch.

[0095] In one embodiment, the composition comprises at least 10 wt% starch, such as at least 15 wt%, such as at least 20 wt%, such as at least 30 wt %, such as at least 40 wt%. In one embodiment, the composition comprises no more than 45 wt% starch, such as no more than 40 wt%, such as no more than 35 wt%, such as no more than 30 wt%, such as no more than 25 wt%, such as no more than 20 wt%. In one embodiment, the composition comprises in the range of 10 to 50 wt% starch, such as 10 to 30 wt%, such as 15 to 25 wt%, such as 10 to 20 wt% starch. In one embodiment, the composition further comprises cellulose. In one embodiment, the cellulose is selected from the group consisting of microcrystalline cellulose (MCC), silicified microcrystalline cellulose and powder cellulose. In one embodiment, the cellulose is MCC.

[0096] In one embodiment, the MCC has a nominal particle size of at least 20 pm, such as at least 50 pm, such as at least 75 pm, such as at least 100 pm, such as at least 125 pm, such as at least 140 pm, such as at least 150 pm, such as at least 160 pm. In one embodiment, the MCC has a nominal particle size of no more than 200 pm, such as no more than 190 pm, such as no more than 180 pm, such as no more than 170 pm, such as no more than 160 pm. In one embodiment, the MCC has a nominal particle size in the range of 140 to 200 pm, such as about 150 to about 180 pm.

[0097] In one embodiment, the composition comprises at least 20 wt% cellulose, such as at least 30 wt%, such as at least 40 wt%, such as at least 50 wt% cellulose. In one embodiment, the composition comprises no more than 60 wt% cellulose, such as no more than 50 wt%, such as no more than 40 wt%, such as no more than 30 wt% cellulose. In one embodiment, the composition comprises in the range of 20 to 60 wt% cellulose, such as 20 to 30 wt%, such as 30 to 40 wt% or such as 50 to 60 wt% cellulose.

[0098] In one embodiment, the composition further comprises lactose or a sugar alcohol. In one embodiment, the lactose is a-lactose. In one embodiment, the lactose is in the form of a monohydrate. In one embodiment, the sugar alcohol is mannitol.

[0099] In one embodiment, the composition comprises in the range of 40 to 70 wt% lactose or sugar alcohol, such as 50 to 60 wt% lactose or sugar alcohol.

[0100] In one embodiment, the composition comprises or consists of GDL, gluconate salt, HPMC, starch, silica and magnesium stearate.

[0101] In one embodiment, the composition comprises or consists of: 10 to 25 wt% GDL; 15 to 30 wt% NaG; 30 to 40 wt% HPMC; 15 to 45 wt% starch; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate. In one embodiment, the composition comprises or consists of: 15 to 25 wt% GDL; 20 to 30 wt% NaG; 30 to 40 wt% HPMC; 15 to 25 wt% starch; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate.

[0102] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0103] In one embodiment, the HPMC is of viscosity type 100 000 mPas. In one embodiment, the HPMC has a degree of substitution of 2208.

[0104] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 30 wt% HPMC; 24.4 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0105] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 35 wt% HPMC; 19.4 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0106] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0107] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19.25 wt% starch; 0.5 wt% silica; and 0.75 wt% magnesium stearate.

[0108] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 18.75 wt% starch; 0.5 wt% silica; and 1.25 wt% magnesium stearate.

[0109] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate. In one embodiment, the composition comprises or consists essentially of about: 10 wt% GDL; 12.25 wt% NaG; 35 wt% HPMC; 41.5 wt% starch; 0.5 wt% silica; and 1.25 wt% magnesium stearate.

[0110] In one embodiment, the composition comprises or consists essentially of about: 10 wt% GDL; 12.25 wt% NaG; 35 wt% HPMC; 41.5 wt% starch; 0.5 wt% silica; and 0.75 wt% magnesium stearate.

[0111] In one embodiment, the composition comprises or consists essentially of about: 15 wt% GDL; 18.375 wt% NaG; 35 wt% HPMC; 30.125 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0112] In one embodiment, the composition comprises or consists of: 15 to 25 wt% GDL; 20 to 30 wt% NaG; 45 to 65 wt% HPMC; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate.

[0113] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 54.4 wt% HPMC; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0114] In one embodiment, the composition comprises or consists of: 15 to 25 wt% GDL; 20 to 30 wt% NaG; 25 to 40 wt% HPMC; 10 to 30 wt% MCC; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate.

[0115] In one embodiment, the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 30 wt% HPMC; 24.4 wt% MCC; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0116] In a preferred embodiment, the composition comprises or consists of essentially about: 15 wt% GDL; 18.4 wt% NaG; 15 wt% HPMC; 14.2 wt% starch; 36 wt% MCC; 0.4 wt% silica; and 1 wt% magnesium stearate.

[0117] In one embodiment, the composition comprises or consists of the compounds as disclosed herein, with the proviso that the sum of the components does not exceed 100 wt%. In a preferred embodiment, the composition comprises or consists essentially of about: 150.0 mg GDL; 184.0 mg NaG; 150.0 mg HPMC; 142.5 mg starch; 360.0 mg MCG; 3.8 mg silica; and 10.0 mg magnesium stearate.

[0118] In one embodiment, the pharmaceutical composition is stable for at least 6 months in 25 °C, or for at least 6 months in 40 °C.

[0119] In one embodiment, the composition does not comprise a further anti-fungal agent. In one embodiment, the anti-fungal agent is selected from the group consisting of miconazole, terconazole, isoconazole, fenticonazole, fluconazole, nystatin, ketoconazole, clotrimazole, butoconazole, econazole, tioconazole, itraconazole, 5- fluoracil, and metronidazole.

[0120] In one embodiment, the composition further comprises a carrier, a filler, and / or a buffering or pH-adjusting agent.

[0121] Subject, individual

[0122] In one embodiment, wherein the subject is a mammal. In one embodiment, the subject is a human. In one embodiment, the subject is a woman.

[0123] In one embodiment, the subject is a pregnant woman. In one embodiment, the subject is a pregnant woman at risk for late miscarriage or pre-term birth.

[0124] In one embodiment, the subject suffers from a microbial infection, such as a urogenital infection.

[0125] In one embodiment, the subject suffers from a microbial infection selected from the group consisting of fungal infections, bacterial infections and mixed fungal and bacterial infections.

[0126] In one embodiment, the fungal infection is a mycosis. In one embodiment, the mycosis is Candidiasis. In one embodiment, the Candidiasis is Candidiasis of vulva and vagina or Candidiasis of urogenital sites. In one embodiment, the bacterial infection is bacterial vaginosis.

[0127] In one embodiment, the fungal infection is caused by a pathogen selected from the group consisting of Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, Trichophyton verrucosum, Trichophyton rubrum, Trichophyton violaceum, Trichophyton tonsurans, Microsporum canis, Malassezia and Aspergillus.

[0128] In one embodiment, the bacterial infection is caused by a pathogen selected from the group consisting of Gardnerella vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum (syphilis), Atopobium vaginae, Prevotella spp, Mobiluncus spp, Peptostreptococcus spp, Poryphyromonas spp, Mycoplasma hominis, Bacteroides spp, Ureaplasma urealyticum, Streptococcus spp, Enterobacteriaceae, Enterococci, Staphylococcus spp, Propionibacterium, Escherichia coli, Klebsiella, Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, Acetinobacter baumanii, Streptococcus pyogenes, Streptococcus agalactiae, Beta- Hemolytic Streptococci Groups C and G and Porphyromonas gingivalis..

[0129] Items

[0130] 1 . A method of increasing growth and / or reproduction of Lactobacillus crispatus in a subject, the method comprising administering to the subject a composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof.

[0131] 2. Use of a composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof, for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

[0132] 3. A composition comprising glucono-delta-lactone (GDL), or a pharmaceutically acceptable salt thereof, for use for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

[0133] 4. An in vitro method of increasing growth and / or reproduction of Lactobacillus crispatus, the method comprising incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising glucono-delta- lactone (GDL),

[0134] 5. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at least 0.75 wt% GDL, such as at least 1 wt%, such as at least 1.5 wt%, such as at least 2 wt%, such as at least 2.5 wt%, such as at least 5 wt%, such as at least 10 wt%, such as at least 15 wt%, such as at least 20 wt%, such as at least 25 wt% GDL.

[0135] 6. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at most 30 wt GDL%, such as at most 35 wt%, such as at most 20 wt%, such as at most 15 wt%, such as at most 10 wt%, such as at most 7.5 wt%, such as at most 5 wt% GDL.

[0136] 7. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises between 1 wt% and 30 wt%, such as between 5 wt% and 20 wt%, such as between 10 wt% and 20 wt%, such as between 1 wt% and 10 wt%, such as between 1 wt% and 7.5 wt% GDL.

[0137] 8. The method, the use or the composition for use according to any one of the preceding items, wherein the increase in growth and / or reproduction is relative to the growth and / or reproduction of L. crispatus prior to incubating Lactobacillus crispatus in a growth medium comprising or consisting of a composition comprising GDL.

[0138] 9. The method, the use or the composition for use according to any one of the preceding items, wherein the increase in growth and / or reproduction is relative to the growth and / or reproduction of L. crispatus in a similar growth medium not comprising or consisting of a composition comprising GDL.

[0139] 10. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is administered to the urogenital system.

[0140] 11. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises a gluconate salt, such as sodium gluconate (NaG).

[0141] 12. The method, the use or the composition for use according to any one of the preceding items, wherein the increase in growth and / or reproduction is relative to the growth and / or reproduction in the subject prior to administration of the composition comprising GDL.

[0142] 13. The method, the use or the composition for use according to any one of the preceding items, wherein the increase in growth and / or reproduction of L. crispatus is measured by measuring the absolute or the mean relative abundance of L. crispatus prior to and after administration of the composition comprising GDL.

[0143] 14. The method, the use or the composition for use according to any one of the preceding items, wherein the growth, such as the level or amount, of L. crispatus is increased at least 1.5-fold, such as at least 2-fold, such as at least 5-fold, such as at least 10-fold, such as at least 20-fold, such as at least 50-fold, such as at least 100-fold, such as at least 500-fold, such as at least 1000-fold, such as at least 5000-fold, such as at least 10,000-fold as compared to the level of L. crispatus prior to administration of the composition comprising GDL.

[0144] 15. The method, the use or the composition for use according to any one of the preceding items, wherein the level of L. crispatus is increased between 1.5-fold and 10,000-fold, such as between 1.5-fold and 10-fold, such as between 1000- fold and 10,000-fold, such as between 4000-fold and 6000-fold, such as between 3000-fold and 7000-fold, such as between 10-fold and 500-fold, such as between 100-fold and 300-fold as compared to the level of L. crispatus prior to administration of the composition comprising GDL.

[0145] 16. The method, the use or the composition for use according to any one of the preceding items, wherein the level of L. crispatus is increased after at least 1 day of administration of the composition comprising GDL, such as after at least 3 days of administration, such as after at least 6 days of administration, such as at least 7 days of administration of the composition comprising GDL.

[0146] 17. The method, the use or the composition for use according to any one the preceding items, wherein the level of L. crispatus remains increased for at least 14 days after administration of the composition comprising GDL, such as for at least 35 days after administration, such as for at least 4 months after administration.

[0147] 18. The method, the use or the composition for use according to any one of the preceding items, wherein the increase in growth, such as the increase in level or amount of L. crispatus, is measured by measuring the mean relative abundance of L. crispatus prior to and after administration of GDL.

[0148] 19. The method, the use or the composition for use according to any one of the preceding items, wherein the mean relative abundance of L. crispatus is at least 10%, such as at least 25%, such as at least 50%, after the final day of administration. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is administered vaginally. The method, the use or the composition for use according to any one of the preceding items, wherein the method, the use, or the composition for use comprises administration of the composition at least once daily, such as at least twice daily, such as at least three times daily. The method, the use or the composition for use according to any one of the preceding items, wherein the method, the use, or the composition for use comprises administration of the composition no more than every second day, such as no more than every third day, such as no more than once a week. The method according to any one of the preceding items, wherein the method, the use, or the composition for use comprises administration of the composition for no more than six days. The method, the use or the composition for use according to any one of the preceding items, wherein the method, the use, or the composition for use comprises administration of the composition for at least one week, such as for at least two weeks, such as for at least three weeks, such as for at least four weeks. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is adapted for administration for: a. at least once daily, such as at least twice daily, such as at least three times daily; and / or b. no more than every second day, such as no more than every third day, such as no more than once a week; and / or c. no more than six days; and / or d. at least one week, such as for at least two weeks, such as for at least three weeks, such as for at least four weeks; and / or e. at least once daily during at least a week. 26. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is administered vaginally once daily for at least six days.

[0149] 27. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises gluconic acid, or a pharmaceutically acceptable salt thereof.

[0150] 28. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises glucono-gamma- lactone, or a pharmaceutically acceptable salt thereof.

[0151] 29. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at least 5 wt %, such as at least 10 wt %, such as at least 15 wt %, such as at least 20 wt %, such as at least 25 wt %, such as at least 30 wt %, such as at least 40 wt %, such as at least 50 wt %, such as at least 60 wt % of GDL.

[0152] 30. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises no more than 99 wt %, such as no more than 95 wt %, such as no more than 90 wt %, such as no more than 85 wt %, such as no more than 80 wt %, such as no more than 75 wt % of GDL.

[0153] 31. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises 5 to 99 wt %, such as in 10 to 95 wt %, such as in 15 to 95 wt %, such as in 20 to 90 wt %, such as in 40 to 95 wt %, such as in 40 to 95 wt %, such as in 50 to 95 wt % of GDL.

[0154] 32. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises between 5 and 25 wt%, such as between 10 and 20 wt%, such as 15 wt% GDL. 33. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises no more than 10 wt % water, preferably no more than 5 wt % water.

[0155] 34. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is formulated as a tampon, vagitorium, vaginal aerosol, vaginal cup, vaginal gel, vaginal insert, vaginal patch, vaginal ring, vaginal sponge, vaginal suppository, vaginal cream, vaginal emulsion, vaginal foam, vaginal lotion, vaginal ointment, vaginal powder, vaginal shampoo, vaginal solution, vaginal spray, vaginal suspension, vaginal tablet, vaginal rod, vaginal disc, vaginal device, and any combination thereof, or wherein the pharmaceutical formulation is present on a sanitary article, such as a tampon, a sanitary napkin, an incontinence pad or diaper, or a panty liner.

[0156] 35. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is formulated as a vagitorium, vaginal insert, vaginal ring, vaginal suppository, vaginal tablet, vaginal rod, or vaginal disc, preferably wherein the composition is formulated as a vaginal tablet.

[0157] 36. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is formulated as a vaginal pessary.

[0158] 37. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is a pharmaceutical formulation.

[0159] 38. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises: a. Glucono-delta-lactone (GDL); b. A gluconate salt, such as sodium gluconate (NaG); and

[0160] 39. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises: a. Glucono-delta-lactone (GDL); and b. Hypromellose (HPMC). The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises: a. Glucono-delta-lactone (GDL); b. A gluconate salt; and c. Hypromellose (HPMC). The method, the use or the composition for use according to any one of the preceding items, wherein the gluconate salt is selected from the group consisting of sodium gluconate, potassium gluconate, calcium gluconate, magnesium gluconate, iron(ll)gluconate, copper(ll)gluconate, and zinc gluconate. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises: a. Glucono-delta-lactone (GDL); b. Sodium gluconate (NaG); and c. Hypromellose (HPMC). The method, the use or the composition for use according to any one of the preceding items, wherein GDL is released for at least 6 h in vivo, such as for at least 12 h, such as for at least 18 h, such as for at least 24 h, such as for at least 36 h, such as for at least 42 h in vivo. The method, the use or the composition for use according to any one of the preceding items, wherein GDL is released for at least 24 h, such as for at least 36 h in vivo. The method, the use or the composition for use according to any one of the preceding items, wherein GDL is released for no more than 72 h, such as for no more than 48 h in vivo. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is in the form of a pharmaceutical dosage form. The method, the use or the composition for use according to any one of the preceding items, wherein the composition is a tablet. The method, the use or the composition for use according to item 47, wherein the mass of the tablet is at least 0.5 g, such as at least 0.7 g, such as at least 0.85 g, such as at least 1.0 g, such as at least 1.25 g, such as at least 1.4 g. The method, the use or the composition for use according to any one of items 47 to 48, wherein the mass of the tablet is no more than 2.0 g, such as no more than 1.75 g, such as no more than 1.5 g, such as no more than 1.0 g, such as no more than 0.9 g. The method, the use or the composition for use according to item 47, wherein the mass of the tablet is in the range of 0.5 to 2.0 g, such as in the range of 0.7 to 1.75 g, such as in the range of 1.4 to 1.6 g, such as about 1.5 g. The method, the use or the composition for use according to item 47, wherein the mass of the tablet is in the range of 0.5 to 2.0 g, such as in the range of 0.5 to 1.25 g, such as in the range of 0.7 to 1.0 g, such as about 0.85 g. The method, the use or the composition for use according to any one of the items 47 to 51 , wherein the tablet is bullet shaped. The method, the use or the composition for use according to any one of the items 47 to 52, wherein the tablet has a width of at least 5 mm, such as at least 7 mm, such as at least 9 mm, such as at least 10 mm. The method, the use or the composition for use according to any one of the items 47 to 53, wherein the tablet has a width of no more than 15 mm, such as no more than 12 mm, such as no more than 11 mm, such as no more than 10 mm. The method, the use or the composition for use according to any one of the items 47 to 54, wherein the tablet has a width in the range of 8 to 12 mm, such as in the range of 8.5 to 11 mm, such as 10 to 10.5 mm. The method, the use or the composition for use according to any one of the items 47 to 55, wherein the tablet has a width in the range of 7.0 to 12 mm, such as in the range of 8.0 to 10 mm, such as 8.5 to 9.5 mm. The method, the use or the composition for use according to any one of the items 47 to 56, wherein the tablet has a length of at least 10 mm, such as at least 15 mm, such as at least 20 mm. The method, the use or the composition for use according to any one of the items 47 to 57, wherein the tablet has a length of no more than 30 mm, such as no more than 25 mm, such as no more than 20 mm. The method, the use or the composition for use according to any one of the items 47 to 58, wherein the tablet has a length in the range of 10 to 30 mm, such as in the range of 15 to 20 mm, such as in the range of 16 to 18 mm. The method, the use or the composition for use according to any one of the items 47 to 59, wherein the tablet has a length in the range of 10 to 30 mm, such as in the range of 22 to 27 mm, such as in the range of 24 to 25 mm. The method, the use or the composition for use according to any one of items 47 to 60, wherein the tablet has a length of 24.5 mm, a width of 10.3 mm and a height of 6.5 mm. The method, the use or the composition for use according to any one of the preceding items, wherein the amount of GDL is 50 to 500 mg, such as 100 to 400 mg, such as 250 to 350 mg, such as about 300 mg. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at least 5 wt% GDL, such as at least 10 wt%, such as at least 15 wt%, such as at least 20 wt% GDL. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises no more than 50 wt% GDL, such as no more than 40 wt%, such as no more than 35 wt%, such as no more than 30 wt%, such as no more than 25 wt%, such as no more than 20 wt% GDL.

[0161] 65. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises in the range of 10 to 30 wt% GDL, such as 15 to 25 wt% GDL, such as about 20 wt% GDL.

[0162] 66. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises a gluconate salt in at least a concentration that corresponds to an equimolar concentration of 10 wt% NaG, such as of at least 15 wt% NaG, such as of at least 20 wt% NaG.

[0163] 67. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at least 10 wt% NaG, such as at least 15 wt% NaG, such as at least 20 wt% NaG.

[0164] 68. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises no more than 30 wt% NaG, such as no more than 25 wt%, such as no more than 20 wt% NaG.

[0165] 69. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises in the range of 10 to 30 wt% NaG, such as 15 to 25 wt%, such as 20 to 25 wt%, such as 24 to 25 wt% NaG.

[0166] 70. The method, the use or the composition for use according to any one of the preceding items, wherein the molar ratio of GDL and gluconate salt is 2: 1 to 1 :2, such as 3:2 to 2:3, such as about 1 :1.

[0167] 71 . The method, the use or the composition for use according to any one of the preceding items, wherein the molar ratio of GDL and NaG is 2: 1 to 1 :2, such as 3:2 to 2:3, such as about 1 :1. 72. The method, the use or the composition for use according to any one of the preceding items, wherein the HPMC is selected from the group consisting of HPMC substitution types 2208, 2910, 1828 and 2906.

[0168] 73. The method, the use or the composition for use according to any one of the preceding items, wherein the HPMC is of substitution type 2208 or 2910.

[0169] 74. The method, the use or the composition for use according to any one of the preceding items, wherein the HPMC has a viscosity of at least 10 mPas, such as at least 15 mPas, such as at least 20 mPas, such as at least 25 mPas, such as at least 30 mPas, such as at least 35 mPas, such as at least 40 mPas, such as at least 45 mPas, such as at least 50 mPas, such as at least 55 mPas, such as at least 60 mPas, such as at least 65 mPas, such as at least 70 mPas, such as at least 75 mPas.

[0170] 75. The method, the use or the composition for use according to any one of the preceding items, wherein the HPMC has a viscosity of no more than 400000 mPas, such as no more than 300 000 mPas, such as no more than 200 000 mPas, such as no more than 150 000 mPas, such as no more than 100 000 mPas, such as no more than 50 000 mPas, such as nomore than 10 000 mPas, such as no more than 1 000 mPas, such as no more than 500 mPas, such as no more than 250 mPas.

[0171] 76. The method, the use or the composition for use according to any one of the preceding items, wherein the HPMC has a viscosity in the range of 50 to 200 mPas, such as about 100 mPas.

[0172] 77. The method, the use or the composition for use according to any one of the preceding items, wherein the HPMC has a viscosity in the range or 50 000 mPas to 200 000 mPas, such as about 100 000 mPas.

[0173] 78. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at least 5 wt% HPMC, such as at least 10 wt%, such as at least 10 wt%, such as at least 15 wt%, such as at least 20 wt%, such as at least 25 wt%, such as at least 30 wt%, such as at least 35 wt%, such as at least 40 wt%, such as at least 45 wt%, such as at least 50 wt% HPMC. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises no more than 60 wt% HPMC, such as no more than 55 wt%, such as no more than 50 wt%, such as no more than 45 wt%, such as no more than 45 wt%, such as no more than 40 wt%, such as no more than 35 wt%, such as no more than 30 wt%, such as no more than 25 wt%, such as no more than 20 wt%, such as no more than 15 wt%, such as no more than 10 wt% HPMC. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises in the range of 5 to 60 wt% HPMC, such as 15 to 40 wt%, such as 20 to 35 wt% HPMC. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises silica. The method, the use or the composition for use according to item 81 , wherein the silica is selected from the group consisting of colloidal anhydrous silica, colloidal silicone dioxide, colloidal hydrated silica and hydrophobic colloidal silica. The method, the use or the composition for use according to any one of items item 81 to 82, wherein the composition comprises in the range of 0.1 to 5 wt% silica, such as 0.1 to 2 wt%, such as about 0.5 wt% silica. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises a lubricant. The method, the use or the composition for use according to item 84, wherein the lubricant is selected from the group consisting of magnesium stearate, calcium stearate, stearic acid, sodium stearylfumarate, glyceryl behenate, glyceryl monooloeate, glyceryl monostearate, glyceryl palmitostearate, sucrose palmitate and sucrose stearate. 86. The method, the use or the composition for use according to any one of items 84 to 85, wherein the lubricant is magnesium stearate.

[0174] 87. The method, the use or the composition for use according to any one of items 84 to 86, wherein the composition comprises in the range of 0.1 to 5 wt% lubricant, such as 0.1 to 2 wt%, such as 0.70 to 1.3 wt%, such as about 1 wt% lubricant.

[0175] 88. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises starch.

[0176] 89. The method, the use or the composition for use according to item 88, wherein the starch is pregelatinised starch.

[0177] 90. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises at least 10 wt% starch, such as at least 15 wt%, such as at least 20 wt%, such as at least 30 wt %, such as at least 40 wt%.

[0178] 91. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises no more than 45 wt% starch, such as no more than 40 wt%, such as no more than 35 wt%, such as no more than 30 wt%, such as no more than 25 wt%, such as no more than 20 wt%.

[0179] 92. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises in the range of 10 to 50 wt% starch, such as 10 to 30 wt%, such as 15 to 25 wt%, such as 10 to 20 wt% starch.

[0180] 93. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises cellulose. 94. The method, the use or the composition for use according to item 93, wherein the cellulose is selected from the group consisting of microcrystalline cellulose (MCC), silicified microcrystalline cellulose and powder cellulose.

[0181] 95. The method, the use or the composition for use according to any one of items 93 to 94, wherein the cellulose is MCC.

[0182] 96. The method, the use or the composition for use according to item 95, wherein the MCC has a nominal particle size of at least 20 pm, such as at least 50 pm, such as at least 75 pm, such as at least 100 pm, such as at least 125 pm, such as at least 140 pm, such as at least 150 pm, such as at least 160 pm.

[0183] 97. The method, the use or the composition for use according to any one of items 95 to 96, wherein the MCC has a nominal particle size of no more than 200 pm, such as no more than 190 pm, such as no more than 180 pm, such as no more than 170 pm, such as no more than 160 pm.

[0184] 98. The method, the use or the composition for use according to any one of items 95 to 97, wherein the MCC has a nominal particle size in the range of 140 to 200 pm, such as about 150 to about 180 pm.

[0185] 99. The method, the use or the composition for use according to any one of items 93 to 98, wherein the composition comprises at least 20 wt% cellulose, such as at least 30 wt%, such as at least 40 wt%, such as at least 50 wt% cellulose.

[0186] 100. The method, the use or the composition for use according to any one of items 93 to 99, wherein the composition comprises no more than 60 wt% cellulose, such as no more than 50 wt%, such as no more than 40 wt%, such as no more than 30 wt% cellulose.

[0187] 101. The method, the use or the composition for use according to any one of items 93 to 100, wherein the composition comprises in the range of 20 to 60 wt% cellulose, such as 20 to 30 wt%, such as 30 to 40 wt% or such as 50 to 60 wt% cellulose. 102. The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises lactose or a sugar alcohol.

[0188] 103. The method, the use or the composition for use according to item 102, wherein the lactose is a-lactose.

[0189] 104. The method, the use or the composition for use according to any one of items 102 to 103, wherein the lactose is in the form of a monohydrate.

[0190] 105. The method, the use or the composition for use according to item 102, wherein the sugar alcohol is mannitol.

[0191] 106. The method, the use or the composition for use according to any one of items 102 to 105, wherein the composition comprises in the range of 40 to 70 wt% lactose or sugar alcohol, such as 50 to 60 wt% lactose or sugar alcohol.

[0192] 107. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists of GDL, gluconate salt, HPMC, starch, silica and magnesium stearate.

[0193] 108. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists of: 10 to 25 wt% GDL; 15 to 30 wt% NaG; 30 to 40 wt% HPMC; 15 to 45 wt% starch; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate.

[0194] 109. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists of: 15 to 25 wt% GDL; 20 to 30 wt% NaG; 30 to 40 wt% HPMC; 15 to 25 wt% starch; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate.

[0195] 110. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate. 111. The method, the use or the composition for use according to item 110, wherein the HPMC is of viscosity type 100000 mPas.

[0196] 112. The method, the use or the composition for use according to any one of items 110 to 111 , wherein the HPMC has a degree of substitution of 2208.

[0197] 113. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 30 wt% HPMC; 24.4 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0198] 114. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 35 wt% HPMC; 19.4 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0199] 115. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate.

[0200] 116. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19.25 wt% starch; 0.5 wt% silica; and 0.75 wt% magnesium stearate.

[0201] 117. The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 18.75 wt% starch; 0.5 wt% silica; and 1.25 wt% magnesium stearate.

[0202] 118. The method, the use or the composition for use according to any one of the preceding items 1 , wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.5 wt% NaG; 35 wt% HPMC; 19 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 10 wt% GDL; 12.25 wt% NaG; 35 wt% HPMC; 41.5 wt% starch; 0.5 wt% silica; and 1.25 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 10 wt% GDL; 12.25 wt% NaG; 35 wt% HPMC; 41.5 wt% starch; 0.5 wt% silica; and 0.75 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 15 wt% GDL; 18.375 wt% NaG; 35 wt% HPMC; 30.125 wt% starch; 0.5 wt% silica; and 1 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists of: 15 to 25 wt% GDL; 20 to 30 wt% NaG; 45 to 65 wt% HPMC; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 54.4 wt% HPMC; 0.5 wt% silica; and 1 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists of: 15 to 25 wt% GDL; 20 to 30 wt% NaG; 25 to 40 wt% HPMC; 10 to 30 wt% MCC; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 20 wt% GDL; 24.1 wt% NaG; 30 wt% HPMC; 24.4 wt% MCG; 0.5 wt% silica; and 1 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, with the proviso that the sum of the components does not exceed 100 wt%. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists of essentially about: 15 wt% GDL; 18.4 wt% NaG; 15 wt% HPMC; 14.2 wt% starch; 36 wt% MCC; 0.4 wt% silica; and 1 wt% magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition comprises or consists essentially of about: 150.0 mg GDL; 184.0 mg NaG; 150.0 mg HPMC; 142.5 mg starch; 360.0 mg MCC; 3.8 mg silica; and 10.0 mg magnesium stearate. . The method, the use or the composition for use according to any one of the preceding items, wherein the pharmaceutical composition is stable for at least 6 months in 25°C, or for at least 6 months in 40°C. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition does not comprise a further antifungal agent. . The method, the use or the composition for use according to item 130, wherein the anti-fungal agent is selected from the group consisting of miconazole, terconazole, isoconazole, fenticonazole, fluconazole, nystatin, ketoconazole, clotrimazole, butoconazole, econazole, tioconazole, itraconazole, 5-fluoracil, and metronidazole. . The method, the use or the composition for use according to any one of the preceding items, wherein the composition further comprises a carrier, a filler, and / or a buffering or pH-adjusting agent. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject is a mammal. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject is a human. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject is a woman. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject is a pregnant woman. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject is a pregnant woman at risk for late miscarriage or pre-term birth. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject suffers from a microbial infection, such as a urogenital infection. . The method, the use or the composition for use according to any one of the preceding items, wherein the subject suffers from a microbial infection selected from the group consisting of fungal infections, bacterial infections and mixed fungal and bacterial infections. . The method, the use or the composition for use according to item 139, wherein the fungal infection is a mycosis. . The method, the use or the composition for use according to item 140, wherein the mycosis is Candidiasis. . The method, the use or the composition for use according to item 141, wherein the Candidiasis is Candidiasis of vulva and vagina or Candidiasis of urogenital sites. 143. The method, the use or the composition for use according to item 139, wherein the bacterial infection is bacterial vaginosis.

[0203] 144. The method, the use or the composition for use according to item 139, wherein the fungal infection is caused by a pathogen selected from the group consisting of Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, Trichophyton verrucosum, Trichophyton rubrum, Trichophyton violaceum, Trichophyton tonsurans, Microsporum canis, Malassezia and Aspergillus.

[0204] 145. The method, the use or the composition for use according to item 139, wherein the bacterial infection is caused by a pathogen selected from the group consisting of Gardnerella vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum (syphilis), Atopobium vaginae, Prevotella spp, Mobiluncus spp, Peptostreptococcus spp, Poryphyromonas spp, Mycoplasma hominis, Bacteroides spp, Ureaplasma urealyticum, Streptococcus spp, Enterobacteriaceae, Enterococci, Staphylococcus spp, Propionibacterium, Escherichia coli, Klebsiella, Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, Acetinobacter baumanii, Streptococcus pyogenes, Streptococcus agalactiae, Beta-Hemolytic Streptococci Groups C and G and Porphyromonas gingivalis..

[0205] Examples

[0206] Example 1 - GDL promotes growth of L. crispatus but not G. vaginalis

[0207] Material and methods

[0208] Pellets of freeze-dried bacteria were resuspended in species-specific broths, plated on agar and cultured in 37° C.

[0209] A single colony from each agar plate was inoculated in the species-specific broth and grown to exponential phase.

[0210] The cultures were thereafter inoculated in 96-well flat-bottom culture plates comprising varying concentrations of GDL + NaG. G. vaginalis cultures were covered with a layer of mineral oil. The plates were covered with a sterile plate sealers and placed in a plate reader at 200 rpm shaking, and the OD600 was read at intervals for 24 hours. Thereafter, the bacterial cultures from were pipetted and a drop was added on the indicator part of a pH-strip to measure pH.

[0211] Results

[0212] The effects on growth of L. crispatus and G. vaginalis treated with GDL was assessed by culturing the bacteria for 24h in aerobic and anaerobic conditions, respectively.

[0213] L. crispatus

[0214] Analyses showed that GDL + NaG promoted the growth of L. crispatus (Figure 1) in media comprising 2,5- 5% GDL + NaG. The pH for the cultures incubated in 2,5- 5% GDL + NaG decreased with increasing concentration of GDL + NaG.

[0215] G. vaginalis

[0216] Analyses showed a clear growth inhibition of G. vaginalis grown for 24 h in the presence of 2,5 and 5% GDL + NaG. The pH of said cultures was decreased compared to other samples, to approximately pH 4 vs pH 7, and slightly lower for 0,5% GDL + NaG.

[0217] Example 2 - Clinical performance in patients with bacterial vaginosis

[0218] The performance of Formulation A tablets in increasing the amount of Lactobacilli in general and Lactobacillus crispatus in particular in human patients with bacterial vaginosis were investigated in a randomized, double-blinded placebo-controlled study. 152 non-pregnant post-menarche, pre-menopausal women with confirmed bacterial vaginosis diagnosis, defined as having at least three out of four of the following criteria (Amsel criteria):

[0219] 1. Thin, white, yellow, homogenous discharge

[0220] 2. Clue cells on microscopy (more than 20 percent of epithelial cells)

[0221] 3. pH of vaginal fluid above 4.5

[0222] 4. Release of fishy odor, “i.e. a positive whiff test” when alkali (10% potassium hydroxide [KOH] solution) is added. On day 0, the patients were included in the study. The patients self-administered Formulation A tablets or a placebo-control tablet once daily for six days (Days 0-5) and a vaginal swab sample for analyses was taken at Day 0 and at Day 7.

[0223] Material and methods

[0224] Formulation A

[0225] Formulation A tablets comprising 150.0 mg GDL; 184.0 mg sodium gluconate (NaG); 150.0 mg hypromellose (HPMC); 142.5 mg starch; 360.0 mg microcrystalline cellulose (MCG); 3.8 mg silica; and 10.0 mg magnesium stearate were prepared according to the following steps: a. GDL, NaG, HPMC, starch and MCC was sieved; b. Silica was mixed with the sieved components of a.; c. Magnesium stearate was mixed with a portion of the mixture of b.; d. The mixture of c. was added to the remaining mixture of b.; and e. The powder mixture in formed d. to tablets.

[0226] Tablets were produced by direct compression in a 10.1 x 17.9 mm punch / die set.

[0227] Vaginal swab procedure

[0228] Vaginal secretion samples were collected by a gynecologist with a kit from DNAGenotek (Omnigene vaginal OMR-130). The sample was taken with the swab which was inserted to the container, the shaft of the swab was snapped off, and the container was closed, with the tip of the swab remaining in the tube. The samples were stored and shipped in -80°C degree freezer until analysed.

[0229] DNA extraction

[0230] Prior to DNA extraction, the samples were thawed and centrifuged and the supernatant was removed. The material was diluted in yeast lysis buffer and incubated at 37°C for 10 min. The samples were centrifuged again and the supernatant was removed. The samples were thereafter extracted according to the MagPure extraction protocol (D6364-02). Purification was performed in a Tecan EVO robot with MagPure’s buffer GW1 as well as 75% ethanol according to the MagPure protocol. The samples were lastly dissolved in EB buffer. qPCR analysis was performed with a validated method, starting with DNA quantification of samples, using the fluorescent nucleic stain Quant-iT™ PicoGreen™ dsDNA reagents and assay kits on Tecan Infinite™ F200. Samples were thereafter transferred to the OpenArray-format, and analysed using the QuantStudio 12K Flex Real-Time PCR System for the targeted organisms. Samples were analysed in triplicates and both negative and positive controls were used.

[0231] Results

[0232] The effects on abundance of Lactobacilli in general and Lactobacillus crispatus in particular in the vaginal swab samples were assessed by qPCR analysis of the vaginal swab samples from patients treated with the Formulation A tablets as compared to patients treated with the placebo tablet, at day 0 and day 7 respectively.

[0233] The abundance of total Lactobacilli increased significantly between day 0 (before treatment) and day 7 (after treatment) for almost all patients treated with tablets of Formulation A (Fig. 3 and Fig. 4).

[0234] In particular, the abundance of L. crispatus increased significantly between day 0 (before treatment) and day 7 (after treatment) for patients treated with Formulation A tablet (Fig. 5). Simultaneously, a significant decrease could be seen in the abundance of G. vaginalis (Fig. 5).

[0235] Further, there is a significant reduction of vaginal dysbiosis (lower CST category) at day 7 (after treatment) as compared to Day 0 (before treatment) in patients treated with Formulation A. For placebo, this shift is not seen (Fig. 6).

[0236] References

[0237] Falconi-McCahill, A., Bacterial Vaginosis: A Clinical Update with a Focus on Complementary and Alternative Therapies. J Midwifery Womens Health, 2019. 64(5): p. 578-591.

[0238] Kalia, N., J. Singh, and M. Kaur, Microbiota in vaginal health and pathogenesis of recurrent vulvovaginal infections: a critical review. Ann Clin Microbiol Antimicrob, 2020. 19(1): p. 5 France, M.T., Ma, B., Gajer, P. et al. VALENCIA: a nearest centroid classification method for vaginal microbial communities based on composition. Microbiome 8, 166

[0239] (2020). htps: / / doi.Org / 10.1186 / S40168-020-00934-6

Claims

Claims1. Use of a composition comprising glucono-delta-lactone (GDL),or a pharmaceutically acceptable salt thereof, for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

2. A composition comprising glucono-delta-lactone (GDL),or a pharmaceutically acceptable salt thereof, for use for increasing growth and / or reproduction of Lactobacillus crispatus in a subject.

3. The use or the composition for use according to any one of the preceding claims, wherein the composition is administered to the urogenital system.

4. The use or the composition for use according to any one of the preceding claims, wherein the increase in growth and / or reproduction is relative to the growth and / or reproduction in the subject prior to administration of the composition comprising GDL.

5. The use or the composition for use according to any one of the preceding claims, wherein the increase in growth and / or reproduction of L. crispatus is measured by measuring the absolute or the mean relative abundance of L. crispatus prior to and after administration of the composition comprising GDL.

6. The use or the composition for use according to any one of the preceding claims, wherein the growth, such as the level or amount, of L. crispatus is increased at least 1.5-fold, such as at least 2-fold, such as at least 5-fold, suchas at least 10-fold, such as at least 20-fold, such as at least 50-fold, such as at least 100-fold, such as at least 500-fold, such as at least 1000-fold, such as at least 5000-fold, such as at least 10,000-fold after the final day of administration as compared to the level of L. crispatus prior to administration of the composition comprising GDL.

7. The use or the composition for use according to any one of the preceding claims, wherein the mean relative abundance of L. crispatus is at least 10%, such as at least 25%, such as at least 50%, after the final day of administration.

8. The method, the use or the composition for use according to any one of the preceding claims, wherein the composition comprises in the range of 1 wt% to 30 wt% GDL, such as 5 wt% to 20 wt%, such as 10 wt% to 20 wt%, such as 15 wt% GDL.

9. The use or the composition for use according to any one of the preceding claims, wherein the composition is administered vaginally.

10. The use or the composition for use according to any one of the preceding claims, wherein the use or the composition for use comprises administration of the composition at least once daily, such as at least twice daily, such as at least three times daily.

11. The use or the composition for use according to any one of the preceding claims, wherein the composition is administered vaginally once daily for at least six days.

12. The use or the composition for use according to any one of the preceding claims, wherein the composition further comprises gluconic acid, or a pharmaceutically acceptable salt thereof.

13. The use or the composition for use according to any one of the preceding claims, wherein the composition further comprises a gluconate salt, such as sodium gluconate (NaG).

14. The use or the composition for use according to any one of the preceding claims, wherein the composition comprises in the range of 10 to 30 wt% NaG, such as 15 to 25 wt%, such as 20 to 25 wt%, such as 24 to 25 wt% NaG.

15. The use or the composition for use according to any one of the preceding claims, wherein the composition further comprises hypromellose (HPMC).

16. The use or the composition for use according to any one of the preceding claims, wherein the composition comprises in the range of 5 to 60 wt% HPMC, such as 15 to 40 wt%, such as 20 to 35 wt% HPMC.

17. The use or the composition for use according to any one of the preceding claims, wherein the composition is a tablet.

18. The use or the composition for use according to claim 17, wherein the amount of GDL in the tablet is 50 to 500 mg, such as 100 to 400 mg, such as 100 to 200 mg, such as about 150 mg.

19. The use or the composition for use according to any one of the preceding claims, wherein the composition comprises or consists of: 10 to 15 wt% GDL;10 to 25 wt% NaG; 10 to 20 wt% HPMC; 30 to 40 wt% MCC; 0.25 to 1.25 wt% silica; and 0.5 to 1.5 wt% magnesium stearate.

20. The use or the composition for use according to any one of the preceding claims, wherein the composition comprises or consists of essentially about: 15 wt% GDL; 18.4 wt% NaG; 15 wt% HPMC; 14.2 wt% starch; 36 wt% MCC; 0.4 wt% silica; and 1 wt% magnesium stearate.

21. The use or the composition for use according to any one of the preceding claims, wherein the composition comprises or consists essentially of about: 150.0 mg GDL; 184.0 mg NaG; 150.0 mg HPMC; 142.5 mg starch; 360.0 mg MCC; 3.8 mg silica; and 10.0 mg magnesium stearate.

22. The use or the composition for use according to any one of the preceding claims, wherein the subject is a mammal.

23. The use or the composition for use according to any one of the preceding claims, wherein the subject is a human.

24. The use or the composition for use according to any one of the preceding claims, wherein the subject is a woman.

25. The use or the composition for use according to any one of the preceding claims, wherein the subject is a pregnant woman.

26. The use or the composition for use according to any one of the preceding claims, wherein the subject is a pregnant woman at risk for late miscarriage or pre-term birth.

27. The use or the composition for use according to any one of the preceding claims, wherein the subject suffers from a microbial infection, such as a urogenital infection.

28. The use or the composition for use according to any one of the preceding claims, wherein the subject suffers from a fungal infection, a bacterial infection or a mixed fungal and bacterial infection.

29. The use or the composition for use according to any one of the preceding claims, wherein the subject suffers from bacterial vaginosis, mycosis, Candidiasis, Candidiasis of vulva and vagina and / or Candidiasis of urogenital sites.

Images