Sponge for vagina

Abstract

The invention relates to a sponge for vagina, which comprises a sponge main body, and the sponge main body is provided with a porous sponge structure. The sponge structure forms at least a portion of the sponge outer surface, and the porous sponge structure is impregnated with a liquid composition comprising one or more bacteriocins. The invention also relates to a liquid composition suitable for use in the vagina, in particular a gel, comprising one or more bacteriocins in an amount that promotes the vaginal microbiota by inhibiting the growth of one or more pathogens. The invention also relates to such a sponge or liquid composition for use in the prevention and / or treatment of dysbiosis, vulvovaginal candidiasis, 10 and / or bacterial vaginosis.

Description

Technical Field

[0001] The present invention relates to a sponge for vaginal use, which can be used as a tampon, wherein the sponge restores the balance of healthy vaginal microflora (dominated by Lactobacillus) by selectively inhibiting pathogens (such as Candida, Staphylococcus aureus, etc.) and retaining Lactobacillus. Background Art

[0002] It is estimated that approximately 75% of women will have a vaginal infection (e.g., candidiasis) at some point in their lives, which may be caused by antibiotic use, use of feminine hygiene products or douching, damp weather, oral contraceptive use, stress, and unknown causes [1]. In approximately 58% of cases, the condition recurs within 12 months. An estimated 8% to 10% of women are susceptible to recurrent VVC (RVVC), with four or more episodes per year [1]. This condition is most commonly caused by Candida albicans, but can also be caused by Candida glabrata, Candida tropicalis, and Candida parapsilosis [2, 3].

[0003] Bacterial vaginosis (BV) is the most common vaginal infection (inflammation) in women of childbearing age and is associated with devastating health problems such as premature birth, risk of human immunodeficiency virus, sexually transmitted disease (STD), and human papillomavirus [4, 5]. The prevalence of BV in the general population is high worldwide, ranging from 23% to 29% per year [6]. BV-related adverse obstetric outcomes range from 8% to 51%. Bacterial vaginosis occurs when Lactobacillus is lost and the microbiota is overgrown with Gardnerella vaginalis and resident anaerobic vaginal bacteria [7].

[0004] Healthy women have an innate vaginal defense mechanism - a vaginal microbiota dominated by Lactobacillus, which regulates the cells of the immune system to prevent infection caused by external pathogens. Multiple lactobacilli predominate in the healthy vagina, and multiple lactobacilli support the defense system and protect against infection by competing for nutrients, preventing adhesion, and secreting antimicrobial compounds (such as bacteriocins, bacteriocin-like substances (such as H 2 O 2 ) and other substances) to inhibit the growth of fungi (e.g., Candida albicans) and pathogenic bacteria [8, 9]. The reduction in the dominance of Lactobacilli leads to various immune changes (e.g., production of pro-inflammatory cytokines / chemokines, increased recruitment of immune cells) and changes in the vaginal lining

[10] .

[0005] Lactobacillus-dominated microbiota can express anti-infective protective factors that inhibit yeast / bacterial growth. Vaginal microbial imbalance (when Lactobacillus is defeated by other bacterial species) is often the cause of gynecological diseases. In general, when the vaginal microbiome is unbalanced, bacterial and fungal infections are promoted. Vaginal infections affect millions of women every year, and these infections usually lead to vaginal dysbiosis-imbalance of the vaginal microbiome often leads to inflammation and / or infection. Therefore, the homeostasis of the microbiome is crucial, especially the need to avoid imbalance of the microbiome or damage to the vaginal mucosa. The vaginal microbiome is a complex ecosystem composed of more than 200 bacterial species, which is influenced by genetic, environmental and behavioral factors, for example

[11] . Vaginal dysbiosis (or lack of Lactobacillus) and infection are even thought to account for 25% to 40% of premature births. Severe deficiency of vaginal Lactobacillus is associated with serious reproductive problems (such as miscarriage and in vitro fertilization (IVF) obstruction)

[12] . Furthermore, the prevalence of abnormal vaginal microbiota is high (30% to 40%) in women undergoing in vitro fertilization

[13] .

[0006] During menstruation, due to the increased pH of the vaginal environment and other factors, lactobacilli are suppressed, resulting in a 100-fold decrease in number, which can then allow pathogens to overgrow [13, 14]. During the rest of the cycle, an increase in bacteria (e.g., Gardnerella vaginalis) is observed in healthy women where lactobacilli dominate, associated with menstruation, accompanied by a decrease in the number of Lactobacillus crispatus and other lactobacilli

[15] . In healthy women, lactobacilli recover after menstruation unless the vaginal environment is compromised by other factors (e.g., stress, medication, hormonal contraception, use of menstrual products). This can lead to dysbiosis and cause infections (e.g., vulvovaginal candidiasis, bacterial vaginosis, BV), and may even lead to premature birth. Medical treatment with antifungal agents or antibiotics cannot restore the balance of the microbiota, and the vicious cycle of recurrent disease continues. Medical treatment is extremely limited, and the inability to prevent frequent, symptomatic recurrences of BV to reduce serious complications (e.g., premature birth) remains a recognized but unresolved problem

[16] . In cases of recurrent candidiasis, treatment is elusive and Candida species evolve with high resistance

[17] .

[0007] Fungal infections (e.g., vulvovaginal candidiasis), bacterial vaginosis, and aerobic vaginosis are the most common categories. Candidiasis is an infection caused by a yeast (a type of fungus) called Candida. Candida can normally reside in the vagina without causing problems. However, if the vaginal environment changes in a way that promotes the growth of Candida, it can multiply and cause infection. Changes in hormone levels, medications, or changes in the immune system can affect the environment.

[0008] In addition, one risk of using tampons is the development of toxic shock syndrome (TSS), which is why some women avoid using tampons. TSS is a rare but life-threatening condition. Up to 99% of these cases are due to using tampons during menstruation. TSS is caused by toxic shock syndrome toxin 1 (TSST-1) produced by Staphylococcus aureus (S. aureus), which can naturally form part of the vaginal microbiota. However, TSS can occur when S. aureus multiplies rapidly. For example, S. aureus growth is promoted when vaginal pH changes and when other adverse conditions are at work. For example, some materials of tampons may promote vaginal colonization or infection more than others

[18] . In particular, high absorbency may lead to toxic shock syndrome, especially because the short, straight, sharp fibers can damage the vaginal mucosa and the toxins produced by S. aureus are rapidly transferred throughout the body. In the UK, 40 cases of this disease are diagnosed each year. Toxic shock syndrome can develop rapidly, and complications may include shock, kidney failure, and death.

[0009] Conventional tampons are made of synthetic materials, polymer foam layers, cotton, or rayon that release dioxins (a pesticide that is harmful to health and the environment) during production and bleaching.

[19] As for cotton tampons, cotton tampons often absorb too much water from the vaginal mucosa (causing excessive drying of the tissue). In addition, cotton tampons are composed of small, sharp fibers that can damage the vaginal mucosa and natural microflora. Such tampons may leave small cotton fragments and old blood particles in the vagina, which can also disrupt the vaginal pH and lead to infection.

[0010] EP 2 448 536A1 describes a compressed tampon, which includes an elongated rod-shaped body with a filler, the elongated rod-shaped body having an insertion end and a withdrawal end, and the withdrawal end has a withdrawal rope. The absorbent body with the filler expands as the fluid is absorbed, and can be made of viscose, cotton or cellulose pulp. The absorbent body with the filler can also include an absorbent sponge or be composed of an absorbent sponge. The tampon also includes a device that is easy to insert, such as an applicator that conforms to the shape of the tampon, and the tampon can be discharged from the applicator. However, this type of tampon is rigid, absorbs excess moisture from the vaginal mucosa, and this type of tampon is composed of small and sharp fibers that damage the vaginal wall and natural microbial flora, thereby leaving small cotton fibers and old blood particles in the vagina. This type of tampon may also disturb the vaginal pH and cause infection, or may cause discomfort due to an uncomfortable shape.

[0011] WO 1992 / 013577A1 discloses a tampon or sanitary napkin for preventing, for example, vaginal infections. Thus, the object of the disclosure is to prevent vaginal infections by introducing a culture of at least one lactic acid-producing bacterium, such as a lactobacillus, into a tampon.

[0012] EP 3 064 072 A1 discloses a composition comprising a bacterial culture of an isolated strain of lactobacillus for use in preventing, for example, candidiasis. However, the disclosure does not relate to a porous sponge structure or to the impregnation of tampons with bacteriocins. Furthermore, when lactobacillus is introduced into the product, it is assumed that the bacteria supports the microflora. However, during menstruation, the vaginal pH level is close to neutral, so that lactobacillus is not in its preferred environment of acidity. Therefore, lactobacillus will not reproduce under these conditions, and the supporting effect on the microflora may be negligible.

[0013] WO 2020 / 003113A1 and US 2021 / 0121596A1 disclose a tampon made of a porous sponge structure of naturally derived glucomannan. The tampon is soft and avoids damage to the vaginal mucosa by being moist, soft and deformable. Summary of the invention

[0014] An object of the present invention is to provide a sponge for the vagina that does not have one or more of the above-mentioned disadvantages. Another object of the present invention is to provide a sponge for the vagina that does not injure and irritate the vaginal wall, does not damage the mucosa, reduces the possibility of infection, facilitates insertion, prevents excessive absorption of body fluids, prevents leakage of menstrual fluids, and / or is convenient during use. Another object of the present invention is to further improve the sponge for the vagina disclosed in WO 2020 / 003113A1 or US 2021 / 0121596A1 by further supporting the natural homeostasis of the vaginal microbiota.

[0015] One or more of the above objects are achieved by impregnating the porous sponge body with a liquid composition comprising one or more bacteriocins. One or more of the above objects are achieved by a liquid composition suitable for vaginal use comprising one or more bacteriocins.

[0016] The present invention provides a sponge for vagina, the sponge comprising a sponge body having a porous sponge structure, as further defined in the claims. The porous sponge body forms at least a portion of the outer surface of the sponge. In addition, the porous sponge body is impregnated with a liquid composition comprising one or more bacteriocins. The present invention also provides a liquid composition suitable for vagina, the liquid composition comprising one or more bacteriocins in an amount that promotes vaginal microflora by inhibiting the growth of one or more pathogens. For example, the one or more pathogens are selected from Staphylococcus aureus, Candida species, Gardnerella vaginalis and combinations thereof. Ideally, the liquid composition is an impregnant for the porous sponge.

[0017] In some embodiments, one or more bacteriocins are synthetic. For example, the bacteriocin can be synthesized in a bacterial strain (e.g., lactobacillus). Alternatively, the bacteriocin can be synthesized by E. coli. For example, nisin can be produced by E. coli or lactobacillus. Alternatively, the bacteriocin can be chemically synthesized. In some embodiments, the sponge does not contain lactobacillus.

[0018] In some embodiments, the sponge is a tampon. For example, the tampon can be a menstrual tampon.

[0019] In some embodiments, the sponge body impregnated with the composition promotes vaginal microbiota during menstruation, during the follicular phase, during the luteal phase, and / or during pregnancy.

[0020] In some embodiments, the composition promotes vaginal microbiota by inhibiting the growth of pathogens, especially Staphylococcus aureus, Candida and / or Gardnerella vaginalis. Ideally, the composition promotes vaginal microbiota by selectively inhibiting the growth of pathogens. In some embodiments, the inhibition ratio of the composition to one or more pathogens is at least 10 times greater than the inhibition ratio of Lactobacillus gasseri and / or Lactobacillus crispatus over a period of 8 hours, particularly, wherein, over a period of 8 hours, the inhibition ratio of the composition to Gardnerella vaginalis is at least 10 times greater than the inhibition ratio of Lactobacillus gasseri and / or Lactobacillus crispatus, more particularly at least 50 times greater, and even more particularly at least 100 times greater. In other words, vaginal microbiota can be promoted by selectively inhibiting pathogens while inhibiting lactobacillus by at least 10 times, at least 50 times, or at least 100 times. In particular, Examples 3 and 4 provide evidence that pathogens are selectively inhibited and lactobacillus is only slightly inhibited or not inhibited.

[0021] In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Staphylococcus aureus compared to culture medium alone under the same conditions. The tampon pouch method can be used to test the growth of Staphylococcus aureus, Candida species and / or Gardnerella vaginalis. The method is disclosed, for example, by Nonfoux et al. (2018)

[20] and is further described in Example 1 herein. In the method, the growth of the pathogen is typically assessed by inserting a tampon / sponge into a sterile plastic bag, adding culture medium (e.g., BHI liquid culture medium) to the bag, inoculating the culture medium with the pathogen, and assessing the growth of the pathogen after incubation. As a positive control in the method, a bag containing an inoculated culture medium without a tampon / sponge is typically used. As used herein, "culture medium alone" refers to a culture medium inoculated with a pathogen (e.g., Staphylococcus aureus, Candida species and / or Gardnerella vaginalis) and without the presence of a porous sponge structure. Otherwise, the conditions used to compare the growth of the pathogen are the same, such as incubation temperature, incubation time, culture medium, and inoculation conditions. By implementing the tampon pouch method, the beneficial effects of the impregnated sponge can be evaluated overall. This is because the combination of the sponge and the impregnation can be compared to the inoculated medium. In some embodiments, the growth of Staphylococcus aureus is inhibited by at least 20 times, at least 50 times, or at least 100 times, or at least 200 times, or at least 350 times after 8 hours of testing by the tampon pouch method.

[0022] In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Candida spp. compared to culture medium alone under the same conditions. In some embodiments, the growth is inhibited by at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 75% after 8 hours of testing by the tampon pouch method.

[0023] In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Gardnerella vaginalis compared to culture medium alone under the same conditions. In some embodiments, the growth of Gardnerella vaginalis is inhibited by at least 50 times, at least 100 times, or at least 250 times, or at least 500 times, or at least 1000 times, or at least 5000 times, or at least 10000 times, or at least 20000 times when tested by tampon pouch method after 24 hours.

[0024] In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Lactobacillus spp. compared to culture medium alone under the same conditions. In some embodiments, the growth of Lactobacillus spp. is inhibited by up to 12-fold when tested by the tampon pouch method after 8 hours.

[0025] In some embodiments, the one or more bacteriocins are selected from class I bacteriocins, class II bacteriocins, class II bacteriocins, and class IV bacteriocins from gram-positive bacteria. In some embodiments, the one or more bacteriocins are selected from class I bacteriocins and / or class II bacteriocins from gram-positive bacteria.

[0026] In some embodiments, the one or more bacteriocins are lantibiotics. In some embodiments, the one or more bacteriocins are type A lantibiotics or type B lantibiotics.

[0027] In some embodiments, the one or more bacteriocins are class II bacteriocins. In some embodiments, the class II bacteriocin is a single-peptide bacteriocin or a double-peptide bacteriocin.

[0028] In some embodiments, the one or more bacteriocins are class IIa bacteriocins. In some embodiments, the class IIa bacteriocins are cyclic bacteriocins.

[0029] In some embodiments, the composition further comprises lactic acid. In some embodiments, the composition comprises an amount of lactic acid such that the pH value of the composition is from 3.5 to 5.5, or from 3.7 to 5, or from 3.9 to 4.5, or about 4.2. In these embodiments, the lactic acid promotes the native lactobacilli in the microbiota by ensuring an acidic pH.

[0030] In certain embodiments, the porous sponge structure is made of natural fibers or synthetic fibers. In particular, the porous sponge structure can be made of natural fibers. In certain embodiments, the porous sponge structure is made of natural polysaccharide-based sponge materials. In certain embodiments, the natural polysaccharide-based sponge materials are glucomannan-based sponge materials. In certain embodiments, the polysaccharide-based materials include glucomannan. In certain embodiments, the polysaccharide-based materials are glucomannan.

[0031] On the other hand, the present invention provides a method for manufacturing a sponge for vagina as disclosed herein, the method comprising the following steps: providing a sponge for vagina, the sponge comprising a sponge body, the sponge body having a porous sponge structure, the porous sponge structure forming at least a portion of the outer surface of the sponge; and impregnating the sponge body with a liquid composition comprising one or more bacteriocins to obtain a sponge for vagina as disclosed herein.

[0032] The present invention also relates to a sponge as disclosed herein or a liquid composition as disclosed, for use in inhibiting the growth of one or more pathogens associated with a condition or disease of the vagina.

[0033] A further aspect of the present invention provides a sponge for vaginal use as disclosed herein for preventing or treating dysbiosis. In addition, the present invention relates to a sponge for preventing or treating vulvovaginal candidiasis and / or bacterial vaginosis. In addition, the present invention relates to a sponge for preventing toxic shock syndrome.

[0034] Furthermore, a liquid composition as described herein for preventing or treating dysbiosis is disclosed. Furthermore, a liquid composition as described herein for preventing or treating vulvovaginal candidiasis and / or bacterial vaginosis is disclosed. Furthermore, a liquid composition as described herein for preventing toxic shock syndrome is disclosed.

[0035] In some embodiments, the liquid composition comprises a fixed amount of one or more bacteriocins. The advantage of a fixed amount is that the amount of available bacteriocins in the vagina does not change. In contrast, if, for example, lactobacilli are incorporated into a sponge, the lactobacilli may produce bacteriocins of different compositions and concentrations. In addition, lactobacilli may produce fewer bacteriocins under adverse conditions, rendering the amount of bacteriocins ineffective. Therefore, impregnating a sponge with a fixed amount of bacteriocins has the advantage that the amount of bacteriocins is always the same and that the technical effect is reproducible. Since the amount of bacteriocins is fixed, the amount of bacteriocins is not affected by the microenvironment.

[0036] In some embodiments, the total concentration of one or more bacteriocins in the liquid composition is from 0.0001% (v / v) to 0.005% (v / v), from 0.0002% (v / v) to 0.002% (v / v), from 0.0004% (v / v) to 0.001% (v / v), from 0.0005% (v / v) to 0.0014% (v / v), from 0.0001% (v / v) to 0.0005% (v / v), or from 0.0014% (v / v) to 0.008% (v / v).

[0037] In some embodiments, the liquid composition is a solution. In some embodiments, the liquid composition is a liquid solution. Typically, a solution is a uniform mixture of two or more substances. In the present case, one or more bacteriocins and other possible ingredients are uniformly mixed to provide a liquid solution.

[0038] In some embodiments, the liquid composition is a gel or lubricant. As known in the art, a gel is a viscous, transparent liquid composition. As also known in the art, lubricants for vaginal use are known in the art, and lubricants are typically water-based or oil-based. In some embodiments, the gel also contains a polysaccharide, such as glucomannan, pectin, inulin, or a combination thereof. Ideally, the gel also contains glucomannan. In some embodiments, the lubricant also contains glucomannan. In particular, Example 4 provides evidence that a gel that additionally contains glucomannan even enhances the growth of lactobacilli while selectively inhibiting pathogens. A gel or lubricant that also contains glucomannan has the following additional advantage, glucomannan also improves the effect of one or more bacteriocins because glucomannan has a prebiotic effect on vaginal lactobacilli. In other words, glucomannan provides "nutrition" for natural lactobacilli, thereby promoting natural lactobacilli. In addition, glucomannan helps to maintain an acidic pH. Therefore, glucomannan also helps, for example, to inhibit vaginal Gardnerella (vaginal Gardnerella is inhibited by acidic pH). Thus, glucomannan promotes natural microflora and / or inhibits pathogens (e.g., Gardnerella vaginalis). At the same time, one or more bacteriocins specifically inhibit pathogenic bacteria and fungi, such as Staphylococcus aureus and Candida. Thus, in these embodiments, both supporting natural lactobacilli by glucomannan and inhibiting pathogens by one or more bacteriocins are achieved.

[0039] The sponge for vagina according to the present invention is impregnated with a liquid composition comprising one or more bacteriocins. A liquid composition comprising said one or more bacteriocins is also disclosed. As shown in Examples 1 and 2, the bacteriocins effectively promote the vaginal microbiota by inhibiting the growth of pathogens (e.g., Staphylococcus aureus, Candida albicans and / or Gardnerella vaginalis). At the same time, the amount of bacteriocins is such that the vaginal microbiota is promoted by inhibiting lactobacilli only to a lesser extent (see Example 1).

[0040] A number of advantages are associated with liquid compositions comprising one or more bacteriocins: the vaginal mucosa is not dried, the natural vaginal pH is maintained as much as possible during menstruation, and the bacteriocins provided support the vaginal microbiota by promoting the microbiota and / or inhibiting the growth of pathogens. In addition, it is advantageous to provide bacteriocins themselves rather than bacteria (e.g., lactobacilli). This is because the production and release of bacteriocins by lactobacilli depends on the optimal growth conditions of lactobacilli, which require, for example, an acidic environment. However, during menstruation, the vaginal pH is close to the pH of blood (pH 7), rather than acidic. Various studies have shown that during menstruation, lactobacilli are reduced by 100 times. During menstruation in healthy women, pathogenic bacteria (e.g., vaginal Gardnerella) increase by 4-log [21, 23]. Therefore, by impregnating a sponge with bacteriocins or by applying a liquid composition comprising one or more bacteriocins, the active agent is directly supplied to the vaginal microbiota. In addition, a specific amount of bacteriocins can be provided, whereas the amount of bacteriocins produced by lactobacilli depends strongly on the environment and is therefore uncontrollable during use. Therefore, the presence of bacteriocins does not depend on the environmental conditions of lactobacilli.

[0041] The combination of a porous sponge structure and impregnation with a liquid composition comprising one or more bacteriocins is particularly advantageous. Impregnation of the porous sponge structure ensures that the bacteriocin is supplied throughout the porous sponge structure. When the porous sponge structure forms at least a portion of the outer surface of the sponge, the liquid composition comprising one or more bacteriocins is in direct contact with the vaginal microbiota, thereby ensuring the supportive effect of the liquid composition. In addition, the porous sponge structure is suitable for absorbing menstrual fluid. The porous sponge structure is suitable for preventing the appearance of fibrils, and is gentle to the vaginal wall, resilient, and shaped to be easily inserted into the vagina.

[0042] Due to the shape and structure of the sponge, liquid is absorbed where it is formed and safely retained within the sponge. Since menstrual blood flows into and is retained in the pores of the sponge, the absorption pattern is fundamentally different from that of conventional tampons, which perform their function when the blood is absorbed by the tampon fabric itself. Menstrual blood accumulated in the pores of the sponge is isolated from the vaginal walls, thereby maintaining a healthier vaginal pH, wherein a healthy vaginal pH (equal to or less than 4.5) is more acidic than blood pH (approximately 7), thereby preserving the healthy microbiome of the vagina and reducing the risk caused by pathogenic bacteria. This effect is further enhanced by the presence of one or more bacteriocins, which further inhibit the growth of pathogens and support natural lactobacilli. The sponge structure enables the sponge to compress and deform during use, ensuring convenience as well as functionality.

[0043] When the sponge is inserted, the sponge regains its shape particularly quickly due to the sponge structure, the material it is made of and the porosity of the sponge. The softness, flexibility and special shape make it easy for the sponge to adapt to the individual physiology without causing discomfort. The sponge is designed to dynamically adapt to the female anatomy during movement, so the sponge is comfortable to wear and effectively absorbs menstrual secretions. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Other objects, principles and features of the present invention will become apparent from the following description, which together with the attached Figure 1 1 and 2 show exemplary embodiments of the present invention. However, it should be understood that these embodiments are not intended to limit the scope of the present invention, but are only used to illustrate exemplary embodiments. In the drawings:

[0045] Figure 1 a, 1b and 1c show a sponge for the vagina without a hollow part according to an embodiment from the front, the side and the top, respectively.

[0046] Figure 2 2a, 2b and 2c show a sponge for the vagina having a hollow portion at a side according to another embodiment from the front, the side and the top, respectively.

[0047] Figure 3 3a, 3b and 3c show a sponge for the vagina having a hollow part at the top according to another embodiment from the front, the side and the top, respectively.

[0048] Figure 4 4a, 4b and 4c show a sponge having a hollow portion at the top according to another embodiment from the top, the side and the top, respectively.

[0049] Figure 5a and Figure 5b The insertion position of the vaginal sponge in the vagina according to two different embodiments of the present invention is schematically shown.

[0050] Figure 6 A sponge for vagina according to still another embodiment is shown in a perspective view.

[0051] Figure 7 Shows the capacity Figure 6 Exemplary packaging of a vaginal sponge.

[0052] Figure 8a Schematically shows the Figure 7 The line AA shown in Figure 6 A cross-sectional view of a sponge for vaginal use.

[0053] Figure 8b Schematically shows the Figure 7 The line BB shown in Figure 6 A cross-sectional view of a sponge for vaginal use.

[0054] Fig. 9 Experimental data are provided for the inhibitory effects of exemplary solutions and gels on Gardnerella vaginalis.

[0055] Fig.10 Experimental data are provided for the effects of exemplary solutions on Lactobacillus gasseri.

[0056] Fig.11a Experimental data on the effect of exemplary gels on Lactobacillus crispatus are provided.

[0057] Fig.11b Experimental data are provided for the effects of exemplary gels on Lactobacillus crispatus and Gardnerella vaginalis. DETAILED DESCRIPTION

[0058] Before providing a detailed description of the preferred embodiments of the present invention with reference to the accompanying drawings, it is noted that like elements are denoted by like reference numerals throughout the drawings.

[0059] It should be understood that many specific details are provided to reveal a complete and understandable description of exemplary embodiments of the present invention. However, it is obvious to those skilled in the art that the details of the embodiments of the present invention do not limit the embodiments of the present invention, and the embodiments of the present invention can be implemented without such specific instructions. Known techniques, processes and components are not described in detail so that the examples of implementing the present invention will not be misleading. In addition, this description should not be considered as limiting the examples presented, but only as embodiments of the examples presented. Any equivalent features or modifications of the features of the present invention are considered to fall within the scope of the present invention.

[0060] Generally, bacteriocins are antimicrobial peptides produced by different bacteria, thereby inhibiting the growth of other bacteria. Bacteriocins can also be effective against yeast. Generally, bacteriocins can kill bacteria or yeast. In particular, bacteriocins are generally more effective for bacteria that do not naturally produce the bacteriocin. By specifically retaining and supporting the natural lactobacillus in the vaginal microbiota with bacteriocins, potential pathogens (such as Candida albicans, Staphylococcus aureus and / or Gardnerella vaginalis) are inhibited from breeding, thereby reducing vaginal infection. This can also prevent recurrence of infection. Ideally, the composition according to the present invention selectively inhibits pathogenic bacteria and fungi (such as Candida albicans, Staphylococcus aureus and / or Gardnerella vaginalis), but protects healthy microbiota (such as Lactobacillus). In this way, the balance of healthy vaginal microbiota is ideally restored and maintained. In particular, the bacteriocin produced by lactobacillus retains natural / vaginal lactobacillus, but inhibits other bacteria and yeast, such as Staphylococcus aureus, Candida albicans and / or Gardnerella vaginalis.

[0061] In some embodiments, one or more bacteriocins are synthetic. Those skilled in the art are aware of commercially available or laboratory-producible synthetic bacteriocins. Synthetic bacteriocins can be chemically synthesized or synthesized in bacterial strains. In the case where the bacteriocin is synthesized in a bacterial strain, the bacteriocin can be separated from the bacterial strain to form a part of the liquid composition. For example, one or more bacteriocins can be synthesized in lactobacillus and then separated from the lactobacillus. In some embodiments, one or more bacteriocins are produced by a bacterial strain. In the case where the bacteriocin is chemically synthesized, the bacteriocin is not produced by a bacterial strain. In addition, the amino acid sequence of the bacteriocin is disclosed in the art, and those skilled in the art are aware of bacteriocin databases that disclose the amino acid sequence of the bacteriocin, such as BAGEL4 (http: / / bagel4.molgenrug.nl / ) or Bactibase (e.g., http: / / bactibase.hammamilab.org / main.php). In addition, products (e.g., tampons) containing bacteria (e.g., lactobacillus) can contain naturally occurring bacteriocins, wherein the bacteriocin is produced by the lactobacillus in the product.

[0062] Regarding liquid compositions suitable for vaginal administration, the liquid compositions include one or more bacteriocins. In particular, one or more bacteriocins are active agents of the liquid compositions. As is well known in the art, an "active agent" can be active. In certain embodiments, the composition is a gel or a lubricant.

[0063] In some embodiments of the invention, the sponge does not contain Lactobacillus. In some embodiments, the sponge does not contain bacteria. In some embodiments, the liquid composition does not contain bacteria (e.g., Lactobacillus).

[0064] In some embodiments, the sponge body impregnated with the composition promotes vaginal microbiota at any stage of the menstrual cycle and / or during pregnancy. Similarly, in some embodiments, the liquid composition promotes vaginal microbiota at any stage of the menstrual cycle and / or during pregnancy. In particular, the sponge body or composition impregnated with the composition can promote vaginal microbiota during menstruation, during the follicular phase, during the luteal phase and / or during pregnancy. Using the sponge during menstruation has the following special advantages: that is, menstrual blood is retained in the pores of the sponge without drying the vaginal mucosa, while providing bacteriocins to the natural lactobacillus of the vagina, thereby inhibiting potential pathogens (such as Staphylococcus aureus, Candida albicans and / or Gardnerella vaginalis). During the follicular phase and the luteal phase, the sponge body or liquid composition impregnated with the composition provides a special advantage of providing one or more bacteriocins to the vaginal microbiota. In particular, recurrent vaginal infections (such as vulvovaginal candidiasis or bacterial vaginosis) can be prevented or treated during the follicular phase and the luteal phase. When the sponge is impregnated with a liquid composition comprising one or more bacteriocins, the sponge is moist and does not dry out the vaginal mucosa. Vaginal infections can recur, particularly during pregnancy. Thus, compared to conventional tampons, the sponge offers the particular advantage of boosting the natural vaginal microflora with bacteriocins while not drying out the mucosa.

[0065] In some embodiments, the sponge is a tampon. In some embodiments, the tampon is a menstrual tampon.

[0066] According to the present invention, the sponge body is impregnated with a liquid composition comprising one or more bacteriocins. As used herein, "impregnation" means that the sponge body is soaked with the liquid composition so that the liquid composition is distributed in the sponge body. For example, the sponge can be made of fibers (e.g., polysaccharide fibers). In this case, the polysaccharide fibers will be impregnated with the liquid composition. If in use, the pores of the porous sponge structure can absorb menstrual fluid, this is advantageous because the sponge will not dry out the vaginal mucosa by retaining menstrual fluid in the pores.

[0067] In some embodiments, the composition promotes vaginal microbiota by inhibiting the growth of one or more pathogens. In some embodiments, the sponge body impregnated with the composition promotes vaginal microbiota by inhibiting the growth of one or more pathogens. By promoting vaginal microbiota, the composition or the sponge body impregnated with the composition has a positive effect on the microbiota. For example, the microbiota is positively affected by promoting the health of the microbiota. For example, the microbiota is positively affected by promoting Lactobacillus. Ideally, the composition or the sponge body impregnated with the composition promotes a balanced vaginal microbiota. The balanced vaginal microbiota is dominated by Lactobacillus. Ideally, the composition or the sponge body impregnated with the composition promotes the dominance of Lactobacillus in the vaginal microbiota. When Lactobacillus is the most in the microbiota, Lactobacillus dominates. Furthermore, when Lactobacillus dominates, no other genus in the microbiota is more than Lactobacillus. Ideally, the health of the microbiota is promoted by the composition or the sponge body impregnated with the composition. In particular, the health of the microbiota can be promoted by promoting the dominance of Lactobacillus. By inhibiting the growth of pathogens, metabolic activity can be changed. For example, the composition or the sponge body impregnated with the composition inhibits the growth of Staphylococcus aureus, Candida and / or Gardnerella vaginalis. In particular, Examples 1 and 2 herein provide data on inhibiting the growth of Staphylococcus aureus, Candida and Gardnerella vaginalis. Advantageously, the composition or the sponge body impregnated with the composition promotes the vaginal microbiota, making it possible to overcome possible imbalances in the vaginal microbiota during, for example, vulvovaginal candidiasis and bacterial vaginosis. More advantageously, the composition or the sponge body impregnated with the composition promotes the vaginal microbiota, making it possible to obtain or maintain a microbiota dominated by Lactobacillus.

[0068] In some embodiments, under the same conditions, the porous sponge structure impregnated with the composition inhibits the growth of Staphylococcus aureus compared to a culture medium alone, wherein the growth is inhibited, for example, by at least 20 times, at least 50 times, or at least 100 times, or at least 200 times, or at least 350 times, as tested by the tampon pouch method after 8 hours. The tampon pouch method is known to those skilled in the art. The method has been described, for example, in Nonfoux L et al. (2018) [7]. In addition, the method is also used in Example 1 and further details are provided. Based on the data disclosed in Example 1, a person skilled in the art is able to test suitable substances and evaluate the inhibition of pathogens (e.g., Staphylococcus aureus, Candida species, and / or Gardnerella vaginalis). Similarly, in some embodiments, under the same conditions, the liquid composition inhibits the growth of Staphylococcus aureus compared to a culture medium alone, wherein the growth is inhibited, for example, by at least 20 times, at least 50 times, or at least 100 times, or at least 200 times, or at least 350 times.

[0069] In some embodiments, under the same conditions, the porous sponge structure impregnated with the composition inhibits the growth of Staphylococcus aureus compared to culture medium alone, wherein the growth is inhibited by at least 100 times after 8 hours as tested by the tampon pouch method. In some embodiments, under the same conditions, the porous sponge structure impregnated with the composition inhibits the growth of Staphylococcus aureus compared to culture medium alone, wherein the growth is inhibited by at least 200 times after 8 hours as tested by the tampon pouch method. In some embodiments, under the same conditions, the porous sponge structure impregnated with the composition inhibits the growth of Staphylococcus aureus compared to culture medium alone, wherein the growth is inhibited by at least 350 times after 8 hours as tested by the tampon pouch method.

[0070] In some embodiments, under the same conditions, the porous sponge structure impregnated with the composition inhibits the growth of Candida compared to a culture medium alone, wherein, after 8 hours, the growth is inhibited by, for example, at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 75% as tested by the sachet method. Similarly, in some embodiments, under the same conditions, the liquid composition inhibits the growth of Candida compared to a culture medium alone, wherein, for example, the growth is inhibited by, for example, at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 75%. In addition, the exemplified data provide further details and demonstrate such growth inhibition. In some embodiments, under the same conditions, the porous sponge structure impregnated with the composition inhibits the growth of Candida compared to a culture medium alone, wherein, after 8 hours, the growth is inhibited by at least 60% as tested by the sachet method. In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Candida compared to culture medium alone under the same conditions, wherein the growth is inhibited by at least 70% after 8 hours as tested by the tampon bag method. In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Candida compared to culture medium alone under the same conditions, wherein the growth is inhibited by at least 80% after 8 hours as tested by the tampon bag method. In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Candida compared to culture medium alone under the same conditions, wherein the growth is inhibited by at least 50%, or at least 60%, or at least 70%, or at least 80% and / or at least 85% after 48 hours as tested by the tampon bag method.

[0071] In some embodiments, after 24 hours, the porous sponge structure impregnated with the composition inhibits the growth of vaginal Gardnerella by at least 50 times, at least 100 times, or at least 250 times, or at least 500 times. Similarly, in some embodiments, under the same conditions, compared with a separate culture medium, the liquid composition inhibits the growth of Candida, wherein the growth is inhibited by, for example, at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 75%. In addition, the data of the examples provide further details and demonstrate the inhibition of this growth. In some embodiments, after 24 hours, the porous sponge structure impregnated with the composition inhibits the growth of vaginal Gardnerella by at least 1000 times. In some embodiments, after 24 hours, the porous sponge structure impregnated with the composition inhibits the growth of vaginal Gardnerella by at least 5000 times. In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of vaginal Gardnerella by at least 10,000 times as tested by the tampon pouch method after 24 hours. In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of vaginal Gardnerella by at least 20,000 times as tested by the tampon pouch method after 24 hours.

[0072] In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Lactobacillus spp. by up to 12-fold compared to culture medium alone under the same conditions, as tested by the tampon pouch method after 8 hours. Up to 12-fold inhibition corresponds to up to 1.079 log 10 In some embodiments, the porous sponge structure impregnated with the composition inhibits the growth of Lactobacillus by up to 12 times, up to 15 times, up to 14 times, or up to 13 times, compared to the culture medium alone, under the same conditions, when tested by the tampon pouch method after 8 hours. Similarly, in some embodiments, the liquid composition inhibits the growth of Lactobacillus by up to 12 times, compared to the culture medium alone, under the same conditions.

[0073] In some embodiments, the one or more bacteriocins are bacteriocins from gram-positive bacteria and / or gram-negative bacteria. In some embodiments, the one or more bacteriocins are bacteriocins from gram-positive bacteria.

[0074] In some embodiments, one or more bacteriocins are selected from class I bacteriocins, class II bacteriocins, class II bacteriocins, and class IV bacteriocins from gram-positive bacteria. For example, one or more bacteriocins can be selected from class I bacteriocins and / or class II bacteriocins from gram-positive bacteria.

[0075] In some embodiments, one or more bacteriocins are bacteriocins from Lactococcus lactis and / or Lactobacillus. In addition, one or more bacteriocins can be from Lactobacillusales. Specifically, Lactobacillusales includes Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, Streptococcus, Aerococcus, Carnosus, Enterococcus, Oenococcus, Tetragenococcus, Rover, Weissella. Alternatively, one or more bacteriocins can be from bacteriocin spore lactobacillus. Bacteriocins derived from lactobacillus have the following special advantages: that is, the bacteriocin has little inhibitory effect on lactobacillus, but has a great effect on other bacteria. For example, one or more bacteriocins can be bacteriocins from Lactobacillus crispatus and / or Lactobacillus gasseri. In some embodiments, one or more bacteriocins are lantibiotics. Lantibiotics are a class of polycyclic peptide antibiotics containing characteristic thioether amino acids lanthionine or methyl lanthionine, as well as unsaturated amino acids dehydroalanine and 2-aminoisobutyric acid. Lantibiotics are produced by Gram-positive bacteria to attack other Gram-positive bacteria (eg, Staphylococcus aureus).

[0076] In particular, one or more bacteriocins can be type A lantibiotics or type B lantibiotics. For example, the lantibiotic can be selected from nisin, lactose, nisin, carnosin, cytolysin, subtilisin, galamycin, epidermin, mesaccharin, actagadine, cinnamycin, duramycin, aramimycin and plant lactobacillus. In some embodiments, the lantibiotic is nisin. In some embodiments, one or more bacteriocins include nisin. In the embodiment where one or more bacteriocins are nisin, nisin can be selected from nisin A, nisin Z, nisin U, nisin Q and nisin F. In some embodiments, nisin is selected from nisin A, nisin Z, or a combination thereof. Ideally, nisin is nisin Z. Alternatively, nisin is nisin A. In some embodiments, the composition comprises a bacteriocin. The bacteriocin may be a nisin, such as nisin A or nisin Z. Ideally, the composition has an acidic pH, such as a pH from 3.5 to 5.5. In some embodiments, the composition comprises a bacteriocin (nisin) and an amount of lactic acid that ensures that the pH of the composition is from 3.5 to 5.5, or from 3.7 to 5, or from 3.9 to 4.5, or about 4.2.

[0077] In some embodiments, one or more bacteriocins are class II bacteriocins. In these embodiments, the class II bacteriocins may be single-peptide bacteriocins or double-peptide bacteriocins. In some embodiments, the class II bacteriocins are single-peptide bacteriocins. In embodiments where one or more bacteriocins are single-peptide bacteriocins, the single-peptide bacteriocins may be selected from pediocin-like bacteriocins and / or non-pediocin-like bacteriocins. In some embodiments, the single-peptide bacteriocin is a non-pediocin-like bacteriocin.

[0078] In some embodiments, the one or more bacteriocins are selected from pediocin PA1, lincomycin A, saxacin P, trevecin A, enterococcin Y105, carnosin BM1, carnosin B2, enterocin A, pierocin 126, bavitacin MN, pierocin V1a, lactococcin A, lactococcin B, biculin 750, lactococcin 972, enterocin B, carnosin A, and capreomycin A. In some embodiments, the one or more bacteriocins are selected from lactococcin A, lactococcin B, biculin 750, lactococcin 972, enterocin B, carnosin A, and capreomycin A. In some embodiments, the one or more bacteriocins are capreomycin A.

[0079] In some embodiments, one or more bacteriocins are class IIa bacteriocins, such as cyclic bacteriocins. In some embodiments, the cyclic bacteriocins are selected from the group consisting of Gacerin A, Rocephin A, butyricin A, lactocyclin Q, cyclin A, lincocyclin Q, amylocyclin A, carnocyclin A, uberrosin A, AS-48, and gavicin ML, preferably, wherein the cyclic bacteriocin is Gacerin A.

[0080] In some embodiments, the composition comprises one, two or three bacteriocins. In some embodiments, the composition comprises one or two bacteriocins. For example, the composition can comprise a Class I and a Class II bacteriocin. An exemplary Class I bacteriocin can be nisin, and an exemplary Class II bacteriocin can be a single peptide bacteriocin, such as capreomycin A. In another example, the composition can comprise a Class I and a cyclic bacteriocin. An exemplary Class I bacteriocin can be nisin, and an exemplary cyclic bacteriocin can be gasterocin A.

[0081] In some embodiments, the pH value of the composition can be from 3.5 to 5.5. In some embodiments, the pH value of the composition can be from 3.7 to 5. The pH value of the composition can be from 3.9 to 4.5. The pH value of the composition can be about 4.2. In some embodiments, the composition also includes lactic acid. For example, the composition can include an amount of lactic acid to bring the pH value of the composition from 3.5 to 5.5. The composition can also include an amount of lactic acid to bring the pH value of the composition from 3.7 to 5. Alternatively, the composition can include an amount of lactic acid to bring the pH value of the composition from 3.9 to 4.5. In some embodiments, the composition can include an amount of lactic acid to bring the pH value of the composition to about 4.2. The lactic acid can be synthetic or can come from a natural source, such as lactic acid bacteria.

[0082] In some embodiments, a person skilled in the art can determine the appropriate concentration of one or more bacteriocins by using the sachet method as disclosed in Examples 1 and 2. In some embodiments, the total concentration of one or more bacteriocins in the liquid composition is from 0.000001% (v / v) to 5% (v / v). Alternatively, the total concentration of the one or more bacteriocins in the composition can be from 0.00001% to 2% of the liquid composition. The total concentration of the one or more bacteriocins in the composition can also be from 0.00005% to 1% of the liquid composition. For example, the total concentration of the one or more bacteriocins in the composition can be from 0.0001% to 0.5% of the liquid composition. Alternatively, the total concentration of the one or more bacteriocins (e.g., capreomycin A) in the composition can be from 0.0005% to 0.25% of the liquid composition. In particular, the total concentration of one or more bacteriocins in the liquid composition can be from 0.0001% (v / v) to 0.005% (v / v). Ideally, the total concentration of one or more bacteriocins (e.g., Gagnerin A) in the liquid composition can be from 0.0002% (v / v) to 0.0020% (v / v). In some embodiments, the total concentration of one or more bacteriocins (e.g., nisin Z) in the liquid composition is from 0.0005% (v / v) to 0.0014% (v / v). In some embodiments, the total concentration of one or more bacteriocins (e.g., nisin A) in the liquid composition is from 0.0005% (v / v) to 0.0014% (v / v). In some embodiments, the total concentration of one or more bacteriocins (e.g., nisin A and / or Z) in the liquid composition is from 0.0006% (v / v) to 0.0013% (v / v). In some embodiments, the total concentration of one or more bacteriocins (e.g., nisin A and / or Z) in the liquid composition is from 0.0007% (v / v) to 0.0012% (v / v). In some embodiments, the total concentration of one or more bacteriocins (e.g., nisin A and / or Z) in the liquid composition is from 0.0008% (v / v) to 0.0011% (v / v). In some embodiments, the total concentration of one or more bacteriocins (e.g., nisin A and / or Z) in the liquid composition is from 0.0009% (v / v) to 0.0010% (v / v). Alternatively, the total concentration of one or more bacteriocins (e.g., nisin A) in the liquid composition can be from 0.0001% (v / v) to 0.0005% (v / v). Ideally, the total concentration of one or more bacteriocins (e.g., nisin) in the liquid composition can be from 0.0014% (v / v) to 0.008% (v / v).

[0083] In some embodiments, the sponge is moist. In these embodiments, the sponge is slightly moist and not dry. The sponge is impregnated with a liquid composition and is therefore moist. The moist sponge is soft and gentle to the vaginal epithelial cells of the mucosa, which keeps the mucosal lining moist and healthy. Because the sponge is soft and therefore easily deformable, the sponge is also comfortable to use.

[0084] The composition can also include one or more excipients. Exemplary excipients are benzalkonium chloride, benzethonium chloride, benzoic acid, benzyl alcohol, boric acid, sodium propionate, sorbic acid, propylene glycol, potassium lactate, calcium acetate, calcium chloride, sodium sorbate, sodium benzoate, sodium chloride, sodium formate, sodium propionate, ethylenediaminetetraacetic acid (EDTA), sodium lactate, organic and inorganic salts and organic and inorganic acids (such as sorbic acid, ascorbic acid, propionic acid and / or fumaric acid). These excipients are kept in the composition by one or more bacteriocins. These excipients also keep sponges. Therefore, excipients have the advantages of stabilizing bacteriocins and keeping sponge materials.

[0085] Those skilled in the art also know suitable buffer solutions for compositions. For example, compositions can also include sodium chloride and EDTA. Alternatively, buffer solutions can also include gelatin phosphate buffer (GPB), sodium phosphate buffer, tris-glycine and tris-tris (hydroxymethyl) methylglycine, octanol phosphate, sodium citrate (BSCSD) combined with sodium diacetate, EDTA, sodium sulfate, sodium phosphate, sodium acetate and sodium citrate, ethylene glycol-bis (β-aminoethyl ether) (EGTA). Compositions can also include benzalkonium chloride, benzethonium chloride, benzoic acid, benzyl alcohol, boric acid, sodium propionate, sorbic acid, propylene glycol, potassium lactate, calcium acetate and / or calcium chloride. For example, the composition may further comprise the following components: benzalkonium chloride at a concentration of 0.01%-0.02% v / v, benzethonium chloride at a concentration of 0.01%-0.02% v / v, benzoic acid at a concentration of 0.1%-0.2% v / v, sodium propionate at a concentration of 5%-10% v / v, and / or sorbic acid at a concentration of 0.05%-0.2% v / v.

[0086] like Figure 1 a to Figure 4 c, an exemplary embodiment of a sponge for vagina 10 includes an absorbent sponge body 12, which includes a porous sponge structure 14. The material of the sponge structure 14 can be a plant-derived material that can be easily deformed and easily re-formed into an undeformed or partially deformed state. This material can be produced from konjac (Amorphophallus) plant root powder by producing glucomannan and then processing the glucomannan by a specific method to obtain a sponge structure, or this material can be made of other naturally occurring polysaccharides (such as but not limited to starch, cellulose, chitosan, chitin and mixtures thereof).

[0087] In some embodiments, the porous sponge structure is made of a natural polysaccharide-based sponge material. The natural polysaccharide-based sponge material can be a glucomannan-based sponge material. In some embodiments, the sponge is made of a completely naturally occurring material (e.g., glucomannan). Due to its biological origin, the glucomannan-based sponge and all its additives are biodegradable and naturally decompose in nature. In these embodiments, the sponge is completely biodegradable and therefore environmentally friendly. In these embodiments, the sponge biodegrades in a natural environment within 30 to 90 days (depending on the environment, humidity level, and number and type of microorganisms). Glucomannan also improves the effect of one or more bacteriocins because glucomannan has a prebiotic effect on vaginal lactobacillus. In other words, glucomannan provides "nutrition" for natural lactobacillus, thereby promoting natural lactobacillus. In addition, glucomannan helps to maintain an acidic pH. Therefore, glucomannan also helps, for example, to inhibit vaginal Gardnerella (vaginal Gardnerella is inhibited by acidic pH). Therefore, glucomannan promotes natural microbiota and / or inhibits pathogens (e.g., vaginal Gardnerella). At the same time, the one or more bacteriocins specifically inhibit pathogenic bacteria and fungi, such as Staphylococcus aureus and Candida. Thus, in these embodiments, both the support of native lactobacilli by glucomannan and the inhibition of pathogens by one or more bacteriocins are achieved.

[0088] Typically, glucomannan is a substance used in the food industry and is also used in wound dressing medicine, which, together with vaginal lactobacilli, inhibits the growth of bacteria such as Staphylococcus aureus and Salmonella typhimurium. The sponge may not contain toxic chemicals and the resulting end product may be biocompatible with the human body, i.e., protection of the natural vaginal microbiota and healthy intimate hygiene is ensured. Glucomannan is hydrophilic and attracts water molecules due to its unique molecular structure, so the sponge wall is always moist and in contact with the vaginal wall through the molecular barrier of the liquid - protecting the extremely sensitive vaginal wall.

[0089] In some embodiments, the porous sponge structure accounts for at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 97%, or at least 99% of the volume of the sponge body. In particular, the porous sponge structure can constitute the entire sponge body. The sponge body can be completely composed of glucomannan.

[0090] According to the present invention, the porous sponge structure forms at least a portion of the outer surface of the sponge. This ensures that the impregnation portion with the liquid composition containing one or more bacteriocins is in direct contact with the vaginal microbiota. This has the advantage that one or more bacteriocins can support natural lactobacilli by inhibiting potential pathogens (such as Staphylococcus aureus and / or Candida). In certain embodiments, the sponge structure forms at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the outer surface of the sponge. In certain embodiments, the porous sponge structure forms 100% of the outer surface of the sponge. This embodiment is particularly advantageous because the surface of the sponge is in contact with the vaginal microbiota to the greatest extent.

[0091] In some embodiments, the sponge includes a sponge body having an insertion end and an extraction end. In some embodiments, the sponge includes a removal facilitator for facilitating removal of the sponge body from a human vagina, the removal facilitator being connected to the sponge body and extending from the extraction end of the sponge body. In addition, the sponge can be placed into the vagina by an applicator.

[0092] In some embodiments, the sponge structure is exposed at least in an area of ​​the outer surface of the body that is located at or near the insertion end of the sponge body. In some embodiments, the outer surface of the body has a fluid capture recess formed at least in part by the exposed portion of the sponge structure. In these embodiments, the fluid capture recess can be positioned closer to the insertion end than the withdrawal end of the sponge body.

[0093] In some embodiments, the sponge body comprises a fluid-impermeable layer covering the sponge structure in a portion of the outer surface of the body. In these embodiments, the fluid-impermeable layer may be made of a non-porous glucomannan-based material.

[0094] exist Figure 1 a to Figure 4 In the embodiment of the present invention, the sponge body 12 further includes a liquid-impermeable layer 16 (i.e., a fluid-impermeable layer; indicated by hatching in the figure), which at least partially covers the sponge structure 14 to keep moisture in the sponge body 12 until the sponge body is used, and prevents liquid from leaking from the sponge body 12 during the use of the sponge body. In some embodiments, the liquid-impermeable layer 16 covers more than half, half, or less than half of the outer surface of the sponge structure 14.

[0095] The sponge may also have an outer fluid barrier layer that provides fluid retention in the sponge even when the sponge deforms during use, i.e., prevents fluid leakage when human movement causes squeezing, bending and similar deformations of the sponge material.

[0096] In some embodiments, the liquid impermeable layer (i.e., fluid impermeable layer) 16 is a non-absorbent, non-porous layer with a thickness of, for example, 0.01 mm to 2 mm. For example, the liquid impermeable layer can be 0.01 mm. It should be understood that the thickness range shown in the liquid impermeable layer 16 is only exemplary and is not intended to limit the scope of the present invention. For example, in other embodiments, the thickness of the liquid impermeable layer 16 can be only a few tenths of a millimeter. The liquid impermeable layer 16 can be made of a glucomannan-based material. The liquid impermeable layer can be directly formed on the sponge structure 14 after the sponge structure 14 has been made. In alternative embodiments, other polysaccharides (e.g., agar, xanthan gum, carrageenan or starch) other than glucomannan can be used as a substrate for making the liquid impermeable layer 16. However, it should be understood that the present invention is not limited to using polysaccharide materials for the liquid impermeable layer 16, and latex or similar polymer materials or resin materials can be used instead.

[0097] In embodiments where the liquid-impermeable layer 16 is omitted, the sponge body 12 may be formed entirely of the sponge structure 14. Thus, the sponge structure 14 may occupy up to 100% of the volume of the sponge body 12. In embodiments with the liquid-impermeable layer 16, the sponge structure 14 may occupy, for example, 90% or more of the volume of the sponge body 12. The remainder of the volume of the sponge body 12 may be formed by the liquid-impermeable layer 16. In some embodiments, the sponge structure 14 occupies at least 95% or at least 97% of the volume of the sponge body 12.

[0098] The sponge may also include an applicator to facilitate insertion and a string, cord, or loop to facilitate removal, but the sponge may also be used without an applicator.

[0099] In some embodiments, the sponge body 12 has an insertion end 18, a withdrawal end 20, and a main body outer surface 22. The insertion end is the front end of the sponge body when the sponge body is pushed into the human vagina, and the withdrawal end is the tail end of the sponge body during the sponge body insertion. The main body outer surface 22 is defined by the outer surface of the sponge body 12. In the case where the sponge structure 14 has one or more exposed portions (i.e., exposed to the outside of the sponge body 12), these exposed portions form at least a portion of the main body outer surface 22. In the case where the liquid-impermeable layer 16 exists and covers at least a portion of the sponge structure 14, the liquid-impermeable layer 16 forms at least a portion of the main body outer surface 22. The extension of the sponge body 12 from the insertion end 18 to the withdrawal end 20 defines the longitudinal extension (or direction) of the sponge body 12. In some drawings, the longitudinal extension is shown by the dotted line 24.

[0100] In some embodiments, the sponge body has a length dimension in the direction from the withdrawal end to the insertion end, wherein the fluid-impermeable layer continuously covers the sponge structure in the following area on the outer surface of the body, which area extends from the withdrawal end over at least one quarter or at least one third or at least half of the length of the sponge body.

[0101] The sponge body 12 may have an insertion end 18, a withdrawal end 20, and a main body outer surface 22. The insertion end is the front end of the sponge body when the sponge body is pushed into the human vagina, and the withdrawal end is the tail end of the sponge body during the sponge body insertion. The main body outer surface 22 is defined by the outer surface of the sponge body 12. In the case where the sponge structure 14 has one or more exposed portions (i.e., exposed to the outside of the sponge body 12), these exposed portions form at least a portion of the main body outer surface 22. In the case where the liquid-impermeable layer 16 exists and covers at least a portion of the sponge structure 14, the liquid-impermeable layer 16 forms at least a portion of the main body outer surface 22. The extension of the sponge body 12 from the insertion end 18 to the withdrawal end 20 defines the longitudinal extension (or direction) of the sponge body 12. In some drawings, the longitudinal extension is shown by a dotted line 24.

[0102] As can be seen in the longitudinal direction 24 from the withdrawal end 20 to the insertion end 18, the sponge body 12 has a lower portion 25, a central portion 26 and an upper portion 28 in sequence. The sponge body has opposite major surfaces 30, 32, each extending between the withdrawal end 20 and the insertion end 18. Considering the expected and appropriate insertion position of the sponge body 12 in the human vagina 34, the major surface 30 is the rear surface of the sponge body 12, and the major surface 32 is the front surface. In this insertion position, the major surface 30 generally faces downward and away from the wearer's uterus 36, and the major surface 32 generally faces upward and toward the uterus 36, as shown in FIG. Figure 5a and Figure 5b As shown. Figure 1 a to Figure 5b In the illustrated embodiment, the major surface 30 has a convex portion in the longitudinal direction 24 substantially over the entire longitudinal length of the sponge body 12. The major surface 32 is generally formed to have a smaller convexity in the longitudinal direction 24 than the major surface 30. In some embodiments (see, for example, Figure 2 b. Figure 3 b. Figure 4 b. Figure 5a , Figure 5b ), when viewed along the longitudinal direction 24, the main surface 32 can be formed to be substantially free of curvature or to have a protrusion in at least a portion of the longitudinal length of the sponge body 12.

[0103] When viewed in a cross-sectional profile (i.e., in a transverse plane), the major surface 30 has a protrusion, wherein the major surface 32 generally has a smaller degree of protrusion than the major surface 30 or has no protrusion at all. The case where there is no protrusion includes that the major surface 32 extends substantially straight in the transverse plane or the profile of the major surface 32 has one or more concave portions. Therefore, when viewed in a cross section orthogonal to the longitudinal direction 24, the sponge body 12 has an irregular, rotationally asymmetric shape. This irregular cross-sectional shape of the sponge body can extend continuously over most of the longitudinal length of the sponge body 12 (e.g., over more than 50%, or more than 60%, or more than 70%, or more than 80%, or more than 90% of the length of the sponge body 12). In some embodiments, the sponge body 12 may also include an outer reinforcing shell (not shown in the figure), which reinforces the structure of the sponge body 12 and helps to maintain the desired shape.

[0104] Exemplary approximate dimensions of the sponge body 12 may be as follows:

[0105] Size 1: Length 4.0cm-4.5cm; Width 1.5cm-2.0cm; Thickness about 1.0cm,

[0106] Size 2: Length 4.5cm; Width 3.6cm; Thickness 1.9cm,

[0107] Size 3: Length 5.8cm; Width 4.5cm; Thickness 2.3cm,

[0108] Size 4: Length 6.0cm; Width 3.9cm; Thickness 2.2cm.

[0109] In some embodiments, the sponge 10 includes a removal facilitator 38, such as a rope, a ring or a line. The removal facilitator 38 is coupled to the sponge body 12 and extends from the withdrawal end 20 in a conventionally known manner per se. The removal facilitator 38 can be attached to the sponge body 12 at both ends on opposite sides of the sponge body 12 approximately in the central portion 26 or the upper portion 28 or the lower portion 25 to form a U-shaped loop, which extends from one side of the sponge body 12 to the other side and forms a possible gap between at least the lower portion 25 of the sponge body 12 and the gripping portion of the removal facilitator 38. The removal facilitator 38 can also be formed as a single element that pierces the sponge body 12 at any position (e.g., the upper portion 28, the central portion 26 or the lower portion 25 of the sponge body 12).

[0110] In some embodiments, the shape of the sponge body 12 is irregular, similar to an oval or any other smooth shape. The sponge body 12 may be generally wider near its insertion end 18 than near its withdrawal end 20. The flat profile mentioned (according to the flat profile mentioned, the width of the sponge body 12 (in Figure 1 a) is greater than the height or thickness of the sponge body (indicated by W in FIG. 1 ) at least over a majority of the longitudinal length of the sponge body 12. Figure 1 b)) is used to reproduce the natural biological vaginal form (https: / / i.pinimg.com / originals / d9 / d1 / 3b / d9d13b90a7d2c5ba6e5314f2c8b3258f.jpg) to create a comfortable experience for the user. This is related to the fact that using a solid form device that produces pressure on the front and back of the vagina (where many nerve endings are concentrated) can cause additional pain or discomfort to the user. Figure 1 a. Figure 2 a. Figure 3 a. Figure 4 As can be seen, the upper portion 28 is generally formed to have a greater width than the lower portion 25. When the sponge body 12 is inserted into the vagina, the sponge body 12 is positioned so that the upper portion 28 of the sponge body is closer to the source of menstrual fluid than the lower portion 25 of the sponge body (see Figure 5a , Figure 5b ). Therefore, the larger width of the upper portion 28 can enhance the absorption of menstrual fluid in the sponge structure 14, and despite the reduced width of the lower portion, the lower portion 28 can effectively retain the accumulated fluid while ensuring high wearing comfort.

[0111] exist Figure 2 a to Figure 5b In the embodiment of the present invention, the sponge body 12 is formed with a hollow portion (recessed portion) 40, which exhibits a generally concave absorption surface 42, where the sponge structure 14 is exposed to the outside of the sponge body 12. If present, the liquid-impermeable layer 16 does not cover the sponge structure 14 at the absorption surface 42. The hollow portion 40 may also be referred to as a fluid capture recess or pocket. In some embodiments, the surface area of ​​the absorption surface 42 is at least 1.5 cm 2 or at least 2.0cm 2 The hollow portion 40 may be concave in all directions, like for example a bowl shape or the like.

[0112] In some embodiments of the present invention, the liquid-impermeable layer 16 may cover the entire outer surface 22 of the sponge body 12 up to the peripheral edge of the hollow part 40. In other embodiments of the present invention, the liquid-impermeable layer 16 may only wrap the sponge structure 14 up to a distance from the peripheral edge of the hollow part 40, so that the sponge structure 14 remains uncovered by the liquid-impermeable layer 16 (i.e., exposed to the outside of the sponge body 12) not only in the area of ​​the hollow part 40 but also outside the hollow part 40 in the area between the liquid-impermeable layer 16 and the peripheral edge of the hollow part 40.

[0113] The hollow portion 40 is designed for the cervix and to facilitate fluid accumulation, and for absorbing fluid faster to prevent blood from stagnating at the cervix. The surface 42 includes a portion of the material of the sponge structure 14 of the sponge body 12, and is generally concave in shape, such as a spoon shape, a hollow hemisphere, an oval shape, or any other form. Although the absorbent surface 42 is referred to as being generally concave, it should be understood that one or more sub-portions of the absorbent surface 42 as follows may still have a planar shape or even a convex shape, and one or more sub-portions of the absorbent surface form a portion of the entire area of ​​the absorbent surface 42. In other words, the absorbent surface 42, while generally forming a concave recess or pocket, may be composed of a concave surface portion and a non-concave surface portion. The hollow portion 40 may be formed on the main surface 32 at a longitudinal position closer to the insertion end 18 than the withdrawal end 20 (e.g., Figure 2 a to Figure 2 c and Figure 5a , Figure 5b ), or may be formed in the region of the insertion end 18 (eg Figure 3 a to Figure 3 c and Figure 4 a to Figure 4 c).

[0114] The sponge body 12 includes at least a non-deformed state when it is fully expanded, a deformed state when it is compressed to occupy the smallest possible volume, and a partially deformed state, in which the sponge body 12 fills the entire vaginal cavity by partially deforming the vaginal cavity wall at the contact site. In this way, the volume of fluid absorbed by the sponge body 12 is increased while ensuring convenient vaginal adaptation. The reversible deformation of the sponge body 12 ensures that the adaptation to the vagina is continuously carried out when the user is at rest and during movement. When the force from the vaginal wall stops acting on the sponge body 12, the deformed sponge body 12 returns to its original shape or part of its original shape.

[0115] from Figure 6 , Figure 7 and Figure 8b It can be seen that the absorbent surface 42 defined by the hollow portion 40 has a concave curved shape in both the width direction and the height direction (the width and height or thickness dimensions are in a plane perpendicular to the longitudinal direction 24) of the sponge body 12. On the side of the main surface 32, the hollow portion 40 forms a deeper neck than on the side of the main surface 30. Therefore, the portion of the hollow portion 40 cut out from the main surface 32 is relatively larger than the portion cut out from the main surface 30.

[0116] In some embodiments, the sponge body 12 generally maintains the shape and volume of the sponge body in an undeformed (i.e., static) state over a wide range of moisture levels of the material of the sponge structure. It is contemplated that, at least for some embodiments, in order to be able to store the sponge 10 for a long period of time without degradation, the material of the sponge structure 14 should be relatively dry rather than relatively wet even when packaged in a sealed manner in a package. Therefore, some embodiments include a capsule or other suitable container (e.g., a pouch) in the package along with the sponge 10, the capsule or container containing a predetermined amount of water or another liquid substance for moistening the sponge body 12 prior to use.

[0117] The embodiment of the present invention also includes a waterproof packaging 44 ( Figure 7 ).exist Figure 7 In the illustrated example, the package 44 is designed as a bag and is configured to maintain sterility and moisture of the body 12 of the sponge 10. It should be understood that many different configurations of the bag are conceivable, and similarly, many other designs of the waterproof package 44 other than the bag are also conceivable, as long as the package 44 is able to maintain a moist and sterile environment for the tampon 10. Figure 6 and Figure 7 The sponge body 12 of the illustrated embodiment has only the sponge structure 14, without the liquid-impermeable layer 16. The package 44 also includes a schematically shown capsule 46 containing, for example, a certain amount of water that the user can use to moisten the sponge body 12 before inserting the sponge body into the human vagina.

[0118] The present invention also relates to a sponge as disclosed herein or a liquid composition as disclosed herein for inhibiting the growth of one or more pathogens associated with a condition or disease of the vagina. Conditions or diseases of the vagina are caused by one or more pathogens, such as bacteria and fungi. Conditions or diseases include, but are not limited to, dysbiosis, bacterial vaginosis, toxic shock syndrome, and vulvovaginal candidiasis.

[0119] For example, a sponge for vagina as disclosed herein is used to prevent or treat vaginal dysbiosis. A liquid composition for vagina as disclosed herein is used to prevent or treat dysbiosis. Dysbiosis (also referred to as dysbacteriosis) is characterized by the destruction of microbial homeostasis caused by an imbalance of the microbial population, a change in the functional composition and metabolic activity of the microbial population, or a change in the local distribution of the microbial population. This dysbiosis may lead to vulvovaginal candidiasis, bacterial vaginosis, and toxic shock syndrome. The present invention also relates to a sponge for vagina as disclosed herein for preventing or treating vulvovaginal candidiasis. The present invention also relates to a liquid composition as disclosed herein for preventing or treating vulvovaginal candidiasis. Vulvovaginal candidiasis can be caused by Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and / or Candida tropicalis. For example, vulvovaginal candidiasis can be caused by Candida albicans. In some embodiments, the vulvovaginal candidiasis is prevented or treated for pregnant women. In some embodiments, the vulvovaginal candidiasis is prevented or treated for menstruating women. The present invention also relates to a sponge for vagina as disclosed herein for preventing or treating bacterial vaginosis. The present invention also relates to a liquid composition as disclosed herein for preventing or treating bacterial vaginosis. Bacterial vaginosis can be caused by Gardnerella vaginalis, Prevotella, Peptostreptococcus, Bacteroides, Leptotrichia and / or Snythia. In some embodiments, the bacterial vaginosis is prevented or treated for pregnant women. In some embodiments, the bacterial vaginosis is prevented or treated for menstruating women. The present invention also relates to a sponge or liquid composition for vagina as disclosed herein for preventing toxic shock syndrome. Toxic shock syndrome can be caused by Staphylococcus aureus. In some embodiments, the toxic shock syndrome is prevented for pregnant women. In some embodiments, the toxic shock syndrome is prevented for menstruating women.

[0120] In some embodiments of the purposes as disclosed herein, the liquid composition inhibits the growth of pathogens (e.g., Staphylococcus aureus, Candida species, and / or Gardnerella vaginalis). In some embodiments, the liquid composition selectively inhibits the growth of one or more pathogens. In these embodiments, the growth of one or more pathogens is strongly inhibited, for example, by at least 10 times, at least 100 times, or at least 1000 times. In some embodiments, within a time period of 8 hours, the liquid composition is at least 10 times greater than the inhibition of Lactobacillus gasseri and / or Lactobacillus crispatus to one or more pathogens. Examples 3 and 4 of the present invention confirm this. In particular, within a time period of 8 hours, the liquid composition is at least 10 times greater than the inhibition of Lactobacillus gasseri and / or Lactobacillus crispatus to vaginalis. For example, the inhibition can be at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 100 times, at least 500 times, or at least 1000 times.

[0121] The sponge and / or liquid composition for vagina according to the present invention advantageously protects the vaginal microbiota and prevents pathogenic infection, particularly during menstruation. As further defined in the claims, the combination of a porous sponge structure impregnated with a liquid composition comprising one or more bacteriocins is advantageous because the combination of the porous sponge structure retains the natural defense mechanism of the vagina, thereby strengthening the natural defense during menstruation. As shown in Examples 1 and 2, the tampon according to the present invention selectively inhibits the growth of Staphylococcus aureus, Candida albicans and / or Gardnerella vaginalis. Therefore, the liquid composition prevents the growth of pathogens, thereby preventing, for example, vulvovaginal candidiasis and / or toxic shock syndrome. In this way, the sponge according to the present invention can also be used for a longer time than a conventional tampon. Due to the inhibitory effect on pathogens, the sponge according to the present invention can also be used for a longer time than a sponge that does not contain a liquid composition with one or more bacteriocins. For example, the sponge according to the present invention can be used for up to 12 hours.

[0122] Although many features and advantages as well as structural details and characteristics of the present invention have been described above, this description is provided as an exemplary embodiment of the present invention. According to the broadest interpretation of the terms used in the claims, changes may be made to the details (especially the form, size and arrangement of materials) without departing from the principles of the present invention.

[0123] The present invention can be further described by the following examples:

[0124] 1. A sponge for vaginal use, comprising:

[0125] The sponge body has a porous structure, and the porous sponge structure forms at least a part of the outer surface of the sponge.

[0126] Therein, the sponge body is impregnated with a liquid composition comprising one or more bacteriocins.

[0127] 2. The sponge according to embodiment 1, wherein the porous sponge structure is made of a polysaccharide-based material comprising glucomannan.

[0128] 3. The sponge of embodiment 1 or 2, wherein the sponge does not contain lactobacilli, in particular, wherein the sponge does not contain bacteria.

[0129] 4. The sponge of any preceding embodiment, wherein the liquid composition comprises a fixed amount of the one or more bacteriocins.

[0130] 5. The sponge of any preceding embodiment, wherein the sponge is moist.

[0131] 6. The sponge of any preceding embodiment, wherein the one or more bacteriocins are synthetic, wherein the one or more bacteriocins are synthesized, for example, in a bacterial strain.

[0132] 4. The sponge of any preceding embodiment, wherein the polysaccharide-based material is glucomannan.

[0133] 5. The sponge of any preceding embodiment, wherein the sponge is a tampon.

[0134] 6. The sponge of any preceding embodiment, wherein the sponge body impregnated with the composition promotes vaginal microbiota during menstruation, during the follicular phase, during the luteal phase, and / or during pregnancy.

[0135] 7. A sponge according to any of the preceding embodiments, wherein the sponge body impregnated with the composition promotes the vaginal microbiota, in particular by inhibiting the growth of pathogens, in particular Staphylococcus aureus, Candida species and / or Gardnerella vaginalis, more particularly, wherein the composition promotes the vaginal microbiota by selectively inhibiting the growth of pathogens, and even more particularly, wherein the composition promotes the vaginal microbiota by selectively not inhibiting the growth of Lactobacillus.

[0136] 8. The sponge of embodiment 7, wherein the sponge body impregnated with the composition inhibits the growth of Staphylococcus aureus compared to culture medium alone under the same conditions, wherein the growth is inhibited, for example, by at least 20 times, or at least 50 times, or at least 100 times, or at least 200 times, or at least 350 times, as tested by the tampon pouch method after 8 hours.

[0137] 9. The sponge of embodiment 7 or 8, wherein the sponge body impregnated with the composition inhibits the growth of Candida spp. compared to culture medium alone under the same conditions, wherein the growth is inhibited by, for example, at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 75% after 8 hours as tested by the tampon pouch method, and by, for example, at least 85% after 48 hours as tested by the tampon pouch method.

[0138] 10. The sponge of any one of embodiments 7 to 9, wherein the porous sponge structure impregnated with the composition inhibits the growth of Gardnerella vaginalis compared to culture medium alone under the same conditions, wherein the growth is inhibited, for example, by at least 50-fold, at least 100-fold, or at least 250-fold, or at least 500-fold, or at least 1000-fold, or at least 5000-fold, or at least 10000-fold, or at least 20000-fold as tested by the tampon pouch method after 24 hours.

[0139] 11. The sponge according to any one of the preceding embodiments, wherein the sponge body impregnated with the composition inhibits the growth of Lactobacillus by up to 12 times compared to the culture medium alone under the same conditions when tested by the tampon pouch method after 8 hours.

[0140] 12. The sponge of any preceding embodiment, wherein the one or more bacteriocins are selected from Class I bacteriocins, Class II bacteriocins, Class II bacteriocins, and Class IV bacteriocins from Gram-positive bacteria, wherein the one or more bacteriocins are, for example, selected from Class I bacteriocins and / or Class II bacteriocins from Gram-positive bacteria.

[0141] 13. The sponge of any preceding embodiment, wherein the one or more bacteriocins are lantibiotics, wherein the one or more bacteriocins are, for example, type A lantibiotics or type B lantibiotics.

[0142] 14. The sponge according to any one of the preceding embodiments, wherein the one or more bacteriocins are class II bacteriocins, wherein the class II bacteriocin is, for example, a monopeptide bacteriocin or a dipeptide bacteriocin.

[0143] 15. The sponge of any preceding embodiment, wherein the one or more bacteriocins are class IIa bacteriocins, wherein class IIa bacteriocins are, for example, cyclic bacteriocins.

[0144] 16. The sponge of any preceding embodiment, wherein the composition further comprises lactic acid, wherein the composition comprises lactic acid, for example, in an amount that ensures the pH of the composition is from 3.5 to 5.5, or from 3.7 to 5, or from 3.9 to 4.5, or about 4.2.

[0145] 17. The sponge of any preceding embodiment, wherein the liquid composition comprises an effective amount of one or more bacteriocins, particularly wherein the effective amount inhibits the growth of Gardnerella vaginalis, Staphylococcus aureus and / or Candida species, more particularly wherein the effective amount selectively inhibits the growth of Gardnerella vaginalis, Staphylococcus aureus and / or Candida species.

[0146] 18. The sponge of any preceding embodiment, wherein the liquid composition comprises a bacteriocin.

[0147] 19. The sponge of any preceding embodiment, wherein the one or more bacteriocins comprises nisin.

[0148] 20. The sponge of embodiment 19, wherein the nisin is nisin A and / or nisin Z.

[0149] 21. The sponge of any preceding embodiment, wherein the liquid composition comprises a bacteriocin, the bacteriocin being nisin, particularly nisin A or nisin Z.

[0150] 22. A method for making a sponge, the method comprising the steps of:

[0151] providing a sponge body, the sponge body having a porous sponge structure, the porous sponge structure forming at least a portion of an outer surface of the sponge; and

[0152] The porous sponge structure is impregnated with a liquid composition comprising one or more bacteriocins to obtain a sponge according to any one of Examples 1 to 21.

[0153] 23. A liquid composition suitable for vaginal administration, the liquid composition comprising one or more bacteriocins in an amount that promotes vaginal microbiota by inhibiting the growth of one or more pathogens.

[0154] 24. The liquid composition of embodiment 23, wherein the pH of the composition is from 3.5 to 5.5.

[0155] 25. The liquid composition of embodiment 23 or 24, wherein the composition does not contain Lactobacillus, in particular, wherein the composition does not contain bacteria.

[0156] 26. The liquid composition of any one of embodiments 23 to 25, wherein the one or more bacteriocins comprises nisin.

[0157] 27. The liquid composition of any one of embodiments 23 to 26, wherein the liquid composition is an impregnating agent for a porous sponge.

[0158] 28. The liquid composition of any one of embodiments 23 to 27, wherein the liquid composition comprises a fixed amount of one or more bacteriocins.

[0159] 29. The liquid composition of any one of embodiments 23 to 28, wherein the liquid composition comprises a bacteriocin.

[0160] 30. The liquid composition of any one of embodiments 23 to 29, wherein the one or more bacteriocins comprises nisin.

[0161] 31. The liquid composition of embodiment 30, wherein the nisin is nisin A and / or nisin Z.

[0162] 32. The liquid composition according to any one of embodiments 23 to 31, wherein the liquid composition comprises a bacteriocin, the bacteriocin being nisin, particularly nisin A or nisin Z.

[0163] 33. The liquid composition of any one of embodiments 23 to 32, wherein the one or more pathogens are selected from Staphylococcus aureus, Candida spp., Gardnerella vaginalis, and combinations thereof.

[0164] 34. The liquid composition of any one of embodiments 23 to 33, wherein the total concentration of the one or more bacteriocins in the liquid composition is from 0.0001% (v / v) to 0.005% (v / v), from 0.0002% (v / v) to 0.002% (v / v), from 0.0004% (v / v) to 0.001% (v / v), from 0.0005% (v / v) to 0.0014% (v / v), from 0.0001% (v / v) to 0.0016 ...2% (v / v) to 0.002% (v / v), from 0.0002% (v / v) to 0.002% (v / v), from 0.0002% (v / v) to 0.002% (v / v), from 0.0002% (v / v) to 0.002% (v / v), from 0.0002% (v / v) to 0.002% (v / v), v / v) to 0.0005% (v / v), or from 0.0014% (v / v) to 0.008% (v / v), in particular, wherein the total concentration of one or more bacteriocins in the liquid composition is from 0.0006% (v / v) to 0.0013%, from 0.0007% (v / v) to 0.0012%, from 0.0008% (v / v) to 0.0011%, or from 0.0009% (v / v) to 0.0010%.

[0165] 35. The liquid composition of any one of embodiments 23 to 34, wherein the liquid composition selectively inhibits the growth of one or more pathogens.

[0166] 36. The liquid composition of any one of embodiments 23 to 35, wherein the liquid composition has at least 10 times greater inhibition of one or more pathogens than Lactobacillus gasseri and / or Lactobacillus crispatus over a period of 8 hours, particularly, wherein the liquid composition has at least 10 times greater inhibition of Gardnerella vaginalis than Lactobacillus gasseri and / or Lactobacillus crispatus over a period of 8 hours, more particularly at least 50 times greater, even more particularly at least 100 times greater inhibition.

[0167] 37. The liquid composition of any one of embodiments 23 to 36, wherein the one or more bacteriocins are further defined in any one of embodiments 12 to 15.

[0168] 38. The liquid composition of any one of embodiments 23 to 37, wherein the composition is a gel, a solution or a lubricant, in particular, wherein the composition is a gel.

[0169] 39. The liquid composition of any one of embodiments 23 to 38, wherein the liquid composition is a gel, and wherein the gel further comprises a polysaccharide, in particular, wherein the polysaccharide is glucomannan.

[0170] 40. The liquid composition of any one of embodiments 23 to 39, wherein the composition further comprises lactic acid, wherein the composition comprises lactic acid, for example, in an amount that ensures that the pH of the composition is from 3.5 to 5.5, or from 3.7 to 5, or from 3.9 to 4.5, or about 4.2.

[0171] 41. The liquid composition of any one of embodiments 23-40, wherein the composition inhibits the growth of one or more pathogens during menstruation, during the follicular phase, during the luteal phase, and / or during pregnancy.

[0172] 42. The sponge of any one of embodiments 1 to 21 or the liquid composition of any one of embodiments 23 to 41 for use in inhibiting the growth of one or more pathogens associated with a condition or disease of the vagina.

[0173] 43. The sponge according to any one of embodiments 1 to 21 or the liquid composition according to any one of embodiments 23 to 41 for use in preventing or treating dysbiosis.

[0174] 44. The sponge of any one of embodiments 1 to 21 or the liquid composition of any one of embodiments 23 to 41 for use in preventing or treating vulvovaginal candidiasis.

[0175] 45. The sponge of any one of embodiments 1 to 21 or the liquid composition of any one of embodiments 23 to 41 for use in preventing or treating bacterial vaginosis.

[0176] 46. ​​The sponge of any one of embodiments 1 to 21 or the liquid composition of any one of embodiments 23 to 41 for use in preventing toxic shock syndrome.

[0177] Example

[0178] The following examples are intended to further illustrate the present invention, but are not intended to limit the present invention. The examples describe the technical features, and the present invention also relates to the combination of the technical features presented in this section.

[0179] Example 1 - Porous sponges impregnated with bacteriocin inhibit the growth of Staphylococcus aureus and Candida albicans

[0180] The purpose of this example is to determine the effect of a test substance on the growth of yeast or bacterial culture. Specifically, in order to identify a beneficial substance for impregnating a porous sponge, various substances were tested for their inhibitory effect on pathogenic bacteria.

[0181] method

[0182] The antifungal effect of menstrual sponges on Candida albicans ATCC 10231 was evaluated. In addition, the antibacterial effect of menstrual sponges on Staphylococcus aureus and Lactobacillus crispatus was evaluated. The tampon pouch method described by Nonfoux L et al. (2018) [7] was used. Briefly, a sample of moistened konjac sponge (glucomannan) (regular size with 6 ml to 9 ml absorbency) was inserted into a 532 ml sterile plastic bag containing 10 ml of brain heart infusion (BHI) liquid culture medium to experiment with Candida ATCC10231 or Lactobacillus crispatus. A sample of moistened konjac sponge (glucomannan) was inserted into a 532 ml sterile plastic bag containing 10 mL of BHI liquid culture medium infused with sheep blood to experiment with Staphylococcus aureus. (BHI and sheep blood were mixed in a 1:1 ratio. The solution was inoculated with 10 5Candida ATCC 10231, Lactobacillus crispatus, Staphylococcus aureus (e.g., Staphylococcus aureus ST20140321) of CFU / ml. Excess air introduced during the insertion of the product into the bag is removed by manual deflation, and the bag is then airtightly sealed. A bag containing inoculated BHI liquid culture medium (without sponge) is used as a positive control. The bag is vertically shaken (about 200rpm) at 37°C and cultivated for 8h or 48h. 4ml solution is sampled from a plastic bag at the end of the cultivation time. Cotton strips are kneaded in the bag for 5 seconds, and liquid is squeezed out and collected by compressing cotton strips for the quantitative determination of bacteria described below. Tryptic soy agar is used to estimate the number of yeast / bacteria in the solution by standard spreading plate technology. After cultivating 8h, 24h and 48h in an aerobic environment at 30°C, colonies are counted.

[0183] result

[0184] Various different types of possible beneficial substances were tested to determine the selective inhibition of pathogenic bacteria or yeast. Surprisingly, bacteriocins were identified as being particularly beneficial for inhibiting pathogens, but not lactobacilli. Natamycin is a known antifungal compound / drug for treating fungal infections around the eyes, for example. Therefore, it can be expected that natamycin will also show antifungal effects in the tampon pouch method. When natamycin from a Streptomyces species was tested for inhibition of Candida albicans by the tampon pouch method, natamycin did not show an inhibitory effect (data not shown).

[0185] Similarly, polylysine has been reported to have bactericidal effects on yeast, fungi, and bacteria. However, when tested using the sachet pouch method, the bactericidal effect on Staphylococcus aureus could not be detected. Specifically, the growth of Staphylococcus aureus was increased from 1.0×10 7 was induced to 2.3×10 8 , and was not inhibited at all. Therefore, although polylysine is expected to have an inhibitory effect on S. aureus, polylysine does not inhibit the growth of S. aureus.

[0186] On the contrary, bacteriocins reliably inhibit the growth of pathogens. Specifically, tampons impregnated with a liquid composition containing nisin and lactic acid at a pH of about 4.2 were tested. The composition reliably inhibited Candida albicans when cultivated for 8 hours, 24 hours, or 48 hours using the tampon pouch method. Specifically, the growth of Candida albicans was evaluated in four separate cultures, and the growth of Candida albicans was reduced by 78%, 75%, 76%, and 73% after 8 hours. In addition, the growth of Candida albicans was reduced by 32%, 35%, 39%, and 28%, respectively, after 24 hours. In addition, the growth of Candida albicans was reduced by 83%, 84%, and 87%, respectively, after 48 hours. In summary, a reduction of about 75% can be observed after 8 hours; a reduction of about 30% can be observed after 24 hours; and a reduction of about 85% can be observed after 48 hours. The percentage represents the number of viable cells (colony counts) compared to control cells grown without a menstrual sponge. The survival rate of the control cells is equivalent to 100%.

[0187] In addition, the bacteriocin also effectively inhibited Staphylococcus aureus. The number of colonies after 8 hours was measured as follows:

[0188]

[0189] On average, the composition is 2.6 log 10 Inhibited Staphylococcus aureus.

[0190] To ensure that the composition did not inhibit vaginal lactobacilli, inhibition of Lactobacillus crispatus was also tested:

[0191]

[0192] On average, the composition inhibited Lactobacillus crispatus only to a small extent, namely 10.95 times, which corresponds to 1.04 log 10 .

[0193] In summary, the tested bacteriocins strongly inhibited the growth of Candida albicans and Staphylococcus aureus, while Lactobacilli were inhibited only to a lesser extent. Thus, tampons impregnated with a liquid composition comprising one or more bacteriocins effectively inhibited Candida albicans and Staphylococcus aureus. In contrast, other tested substances previously reported to have an antibacterial or antifungal effect did not have such an effect.

[0194] Example 2 - Porous sponges impregnated with bacteriocin contain no bacteria and inhibit the growth of Gardnerella vaginalis

[0195] The purpose of this example is to verify that the impregnated porous sponge does not contain bacteria. Additionally, this example tests the inhibition of the growth of Gardnerella vaginalis.

[0196] Materials and methods

[0197] Bacterial strain: Gardnerella vaginalis ATCC 49145.

[0198] Liquid medium and incubation method: BHI (sterile brain heart infusion, Liofilchem, No. 620008) solution supplemented with 1% D-glucose and 2% horse serum (Thermofisher Scientific, Oxoid, No. SR 0035). The inoculated medium was incubated at 37°C in a 50 ml conical centrifuge tube (Thermofisher Scientific, Nunc) in a vertical position without shaking. During the incubation, the tube was gently rotated several times. This method is considered equivalent to the sliver bag method.

[0199] Solid medium: sterile chocolate agar with Vitox plates (Thermo Fisher Scientific, Oak Oak, No. PO5090A). 2 5% CO provided by Gen 2.5L (Thermo Fisher Scientific, Oak Oak, No. CD0025A) 2 The cells were incubated at 37°C in an atmosphere for 43 hours.

[0200] a) Porous sponges impregnated with bacteriocin contain no bacteria

[0201] While other products on the market may contain bacteriocins produced by bacteria within the product, the present invention relates to a porous sponge impregnated with a liquid composition comprising bacteriocins. Ideally, the porous sponge does not contain bacteria.

[0202] To verify that the porous sponge does not contain bacteria (e.g., lactobacilli), three porous sponges (wet konjac sponges made of glucomannan) were incubated in growth medium and tested for potential bacterial growth by counting potential colony forming units (CFU). The tested sponges were impregnated with a liquid composition comprising nisin and lactic acid at a pH of about 4.2.

[0203] Specifically, three sponges were inserted into 50 ml sterile tubes, respectively. 10 ml of liquid culture medium was added to each tube. The control tube contained only culture medium. After 24 hours of cultivation at 37°C, the sponge was compressed several times using a sterile pipette tip. 200 μL of culture medium was taken out from each tube and 200 μL of culture medium was spread on a culture plate. The plates were evaluated after cultivation. This method is considered to be equivalent to the tampon pouch method.

[0204] All the plates were found to be free of microbial colonies. Hence, the sponges tested were not contaminated with bacteria. It can therefore be concluded that the porous sponges tested did not contain bacteria.

[0205] b) Porous sponges impregnated with bacteriocin inhibit the growth of Gardnerella vaginalis

[0206] Since Gardnerella vaginalis is the most common cause of bacterial vaginosis, the inventors also tested the inhibitory effect of the porous sponge on the growth of Gardnerella vaginalis.

[0207] Two independent tests (A, B) were performed. Each test compared bacterial growth in the presence and absence of a sponge. A porous sponge (a moistened konjac sponge made of glucomannan impregnated with a liquid composition containing nisin and lactic acid at a pH of about 4.2) was inserted into the bottom of a 50 ml conical centrifuge tube. Gardnerella vaginalis ATCC 49145 was recovered from frozen storage by streaking and incubating on plates and the OD was measured. 600 The optical density of 0.12 was suspended in liquid culture medium (Test A, Test B). During the preliminary test, the density 100 times and 10 times lower did not provide sufficient bacterial growth.

[0208] Pour 15 ml of bacterial suspension onto the sponge and 15 ml of bacterial suspension into the control tube. Use a serological pipette to evenly wet the sponge.

[0209] Place the tube under 5% CO 2 The sponges were loosely twisted (A) or tightly twisted (B) in air and incubated at 37°C for 24 hours (Test A, Test B). Due to the known slow growth rate of G. vaginalis, 8 hours of incubation were not considered for the evaluation (Janulaitiene, 2018, Figure B in S2 File). After 24 hours of incubation, the sponge was compressed ten times in the tube using a sterile syringe barrel. The OD of the suspension was measured. 600 0.5 ml of culture medium was removed from each tube and serially diluted 10-fold with sterile PBS until the factor was 10. -7 100 μl of the suspension of the relevant dilution was plated in duplicate. Colonies were counted manually on a plate suitable for reading. The counts were used to calculate the colony forming units per milliliter (CFU / ml) of the undiluted liquid culture. The bacterial growth inhibition coefficient was calculated by dividing the CFU / ml of the culture without sponge by the CFU / ml of the sponge-grown culture.

[0210] result:

[0211] (A)

[0212]

[0213] (B)

[0214]

[0215]

[0216] In these tests, the OD of cultures without sponges was 600 The sufficient growth potential of the starting material G. vaginalis suspension was demonstrated. 600 As shown, the tubes with sponges indeed showed significantly less bacterial growth. In summary, the porous sponges impregnated with the liquid composition containing bacteriocin showed a growth rate of 3.6×10 4 and 2.6×10 6 The growth of Gardnerella vaginalis ATCC49145 was inhibited by a coefficient of .

[0217] Example 3 - Solutions containing bacteriocins have an inhibitory effect on Gardnerella vaginalis

[0218] The purpose of this example was to test the inhibitory effect of a composition containing a bacteriocin on Gardnerella vaginalis, the main cause of bacterial vaginosis.

[0219] Materials and methods

[0220] A liquid solution containing nisin (also called "Avodes solution") was tested in this example. In addition, the composition contained lactic acid with a pH of about 4.2. The bacterial strain used in this example was Gardnerella vaginalis ATCC 49145. The culture medium was a liquid medium BHI (brain heart infusion) solution (Levitra) supplemented with horse serum (Thermo Fisher Scientific, Oak Oak). As a solid culture medium, sterile chocolate agar (Thermo Fisher Scientific, Oak Oak) was used. All experiments were performed at 37°C with CO 2 The experiment was carried out under anaerobic conditions provided by Reliance 2.5 (Thermo Fisher Scientific, Oak Oak, OH) to resemble the conditions in the vagina as much as possible.

[0221] Mix 10 mL of liquid culture medium with 6 mL of avodice solution in a sterile glass tube. Use a final concentration of 4-6 × 10 5 Each tube was inoculated with 100 CFU / ml of vaginal Gardnerella. As controls, (i) a sterile glass tube with 10 mL of liquid culture medium (negative control) and no bacteria, (ii) a sterile glass tube with 10 mL of liquid culture medium inoculated with 4-6 × 10 5CFU / ml of vaginal Gardnerella (positive control), and (iii) a sterile 10 mL glass tube mixed with 6 mL of solution (control solution). After 8 hours and 24 hours of cultivation, 100 μL of culture medium was taken out from each tube, 100 μL of culture medium was diluted (if necessary), and 100 μL of culture medium was spread on the culture plate. The results were evaluated after 48 hours of cultivation. The solutions provided were sterile (negative control plates had no microbial colonies).

[0222]

[0223] result

[0224] The experiment showed that despite several hours of incubation at 37°C in a growth medium, the "Avodice solution" had a significant inhibitory effect on the bacterial growth of vaginal Gardnerella. While the number of CFU increased by about 50 times after 8 hours in the control solution, the number of CFU decreased by about 2 times after 8 hours in the "Avodice solution". The effect was even stronger after 24 hours of incubation: while the number of CFU increased by about 7230 times after 24 hours in the control solution, the number of CFU increased by only about 22 times after 24 hours in the "Avodice solution". The bacterial growth inhibition coefficient reached 1.02×10 after 8 hours. 2 , reaching 3.18×10 after 24 hours. 4 .

[0225] These are very promising results, indicating that compositions comprising bacteriocins effectively inhibit the growth of Gardnerella vaginalis, a major cause of bacterial vaginosis. Since Gardnerella vaginalis is typically found in low numbers in the vaginal macrobiota, compositions comprising bacteriocins (e.g., nisin) are expected to effectively keep the number of Gardnerella vaginalis bacteria low and / or even reduce the number of Gardnerella vaginalis bacteria. Importantly, the exemplary solution can strongly inhibit Gardnerella vaginalis while only very mildly inhibiting bacteria that promote the vaginal microbiota (e.g., Lactobacillus crispatus or Lactobacillus gasseri (see also next example)).

[0226] Example 4 - Solutions and gels containing bacteriocins have strong inhibitory effects on Gardnerella vaginalis and only slight inhibitory effects on Lactobacillus

[0227] The purpose of this example was to test the inhibitory effect of solutions and gels containing bacteriocins on Gardnerella vaginalis and Lactobacillus. Since Gardnerella vaginalis is the main cause of bacterial vaginosis, a strong inhibitory effect would be advantageous. In contrast, Lactobacillus supports the vaginal microbiota, so it is advantageous if Lactobacillus is not inhibited or only slightly inhibited. The potential selective effect of the solutions and gels was also tested.

[0228] Materials and methods

[0229] In this example, a liquid solution containing nisin (also referred to as "Avodice solution") was tested. In addition, a gel containing nisin (also referred to as "Avodice gel") was tested in this example. Avodice gel additionally contained glucomannan. In addition, both compositions (solution and gel) contained lactic acid and had a pH value of about 4.2. The bacterial strains used in this example were Gardnerella vaginalis ATCC 49145, Lactobacillus crispatus DSM 20584, and Lactobacillus gasseri DSM20243.

[0230] Culture conditions: BHI (brain heart infusion, Sigma-Aldrich) supplemented with 1% D-glucose and 2% horse serum (Thermo Fisher Scientific, Oak Oak) was used to grow Gardnerella vaginalis. The inoculated culture was incubated at 37°C in a sterile 15 ml conical centrifuge tube in a vertical position without shaking. To determine the number of CFU, plates were plated on chocolate agar with Vitaxel plates (Thermo Fisher Scientific, Oak Oak) with CO 2 Dependent bacteria TM The inoculated culture medium was incubated at 37°C in a sterile 15 ml conical centrifuge tube in a vertical position without shaking and heated with CO 2 Dependent bacteria TM Compact Bag (Thermo Fisher Scientific, Oak Oak) was grown on MRS agar with Tween 80 (Bio-La Roche).

[0231] 0.5 ml of Avodice solution and 0.5 ml of Avodice gel were diluted with 1 ml of MRS or 1 ml of BHI culture medium in a 24-well plate and incubated for 24 hours at 37°C. The control group contained only the corresponding culture medium. 200 μl of culture medium was taken out from each well and 200 μL of culture medium was spread on the culture plate. The plate was evaluated after incubation.

[0232] To evaluate the inhibitory effect on vaginal G. vaginalis, two independent tests (Test A, Test B) were performed with Avodice solution and Avodice gel and then compared with a control group containing the corresponding medium and sodium lactate and lactic acid to maintain a pH of 4.2. 1 ml of Avodice solution was transferred to a 24-well plate and mixed with 1 mL of vaginal G. vaginalis inoculum in BHI liquid medium supplemented with 1% D-glucose and 2% horse serum to a final concentration of 10 5 CFU / ml. The plates were maintained under anaerobic conditions with CO2 Dependent bacteria TM The compact bags were incubated at 37°C for 8 and 24 hours. After incubation, the cultures were serially diluted 10-fold on chocolate agar with sterile PBS until a factor of 10 was reached. -9 The colonies were counted manually on plates suitable for reading. The counts were used to calculate colony forming units per milliliter (CFU / ml) of undiluted liquid culture.

[0233] result:

[0234]

[0235] These results show that the composition (solution or gel) significantly inhibited the growth of G. vaginalis. Therefore, these results further confirm the data of the previous examples. In addition, the data show that the gel is even more beneficial, as the inhibitory effect is even greater, reaching 2.01×10 after 8 hours. 4 times, reaching 3.29×10 after 24 hours. 5 In particular, this indicates that the gel containing nisin strongly inhibited the growth of vaginal Gardnerella. The data reflected in the above table are also Fig. 9 Shown in.

[0236]

[0237] Evaluation of Growth Inhibition of Lactobacillus gasseri

[0238] To evaluate the inhibitory effect on Lactobacillus gasseri, two independent tests (Test A, Test B) were performed with Avodice solution and Avodice gel and then compared with a control group containing the corresponding medium and sodium lactate and lactic acid to maintain a pH of 4.2. 1 ml of Avodice solution was transferred to a 24-well plate and mixed with 1 ml of Lactobacillus gasseri inoculum in MRS liquid medium to a final concentration of 10 5 CFU / ml. The plates were maintained under anaerobic conditions with CO 2 Dependent bacteria TM The compact bags were incubated at 37°C for 8 and 24 hours. After incubation, the cultures were serially diluted 10-fold on chocolate agar with sterile PBS until a factor of 10 was reached. -9 The colonies were counted manually on plates suitable for reading. The counts were used to calculate colony forming units per milliliter (CFU / ml) of undiluted liquid culture.

[0239] result:

[0240]

[0241] The data show that the Avodice solution only very slightly inhibits lactobacilli (e.g., Lactobacillus gasseri). Since, for example, Lactobacillus gasseri supports a healthy microbiome of the vagina, it is particularly advantageous that the solution only slightly inhibits lactobacilli while very strongly inhibiting Gardnerella vaginalis. This selective inhibition is surprising and was not expected.

[0242] Evaluation of Growth Inhibition of Lactobacillus crispatus

[0243] To evaluate the effect on Lactobacillus crispatus, two independent tests (Test A, Test B) were performed with Avodice solution and Avodice gel and then compared with a control containing culture medium with sodium lactate and lactic acid to maintain a pH of 4.2. 1 ml of Avodice solution was transferred to a 24-well plate and mixed with 1 ml of Lactobacillus gasseri inoculum in MRS liquid medium to a final concentration of 10 5 CFU / ml. The plates were maintained under anaerobic conditions with CO 2 Dependent bacteria TM The compact bags were incubated at 37°C for 8 and 24 hours. After incubation, the cultures were serially diluted 10-fold on chocolate agar with sterile PBS until a factor of 10 was reached. -9 The colonies were counted manually on plates suitable for reading. The counts were used to calculate colony forming units per milliliter (CFU / ml) of undiluted liquid culture.

[0244] result:

[0245]

[0246]

[0247] The data further confirm that not only Lactobacillus gasseri, but also Lactobacillus crispatus is only very slightly inhibited or not inhibited at all by the composition comprising bacteriocin. In particular, the gel does not inhibit at all or only very slightly inhibits both tested lactobacilli. In summary, the data show that the composition (solution or gel) selectively inhibits bacteria: the composition strongly inhibits Gardnerella vaginalis and only very slightly inhibits Lactobacillus. In addition, the gel even increases the growth of Lactobacillus crispatus by at least one logarithmic unit, i.e. at least 10 times. This selective inhibition is expected to support the natural microflora of the vagina. Due to this strong effect, this selective inhibition is also expected to prevent bacterial vaginosis, candidiasis and / or dysbiosis. Due to this strong effect, this selective inhibition is also expected to treat bacterial vaginosis, candidiasis and / or dysbiosis.

[0248] References

[0249] 1. J.Yano, J.D.Sobel et al., "Current patient perspectives of vulvovaginal candidiasis: incidence, symptoms, management and posttreatment outcomes", March 29, 2019; 19(1): 48, doi:10.1186 / s12905-019-0748-8.

[0250] 2. F.Bernardis et al., "Candida vaginitis: virulence, host response and vaccine prospects", Med Mycol, April 1, 2018; 56(Suppl_1): 26-31, doi:10.1093 / mmy / myx139.

[0251] 3. J.D.Sobel, "Vulvovaginal candidiasis: epidemiologic, diagnostic, and therapeutic considerations", Am J Obstet Gynecol, February 1998; 178(2): 203-11, doi:10.1016 / s0002-9378(98)80001-x.

[0252] 4. E.Gillet et al., "Bacterial vaginosis is associated with uterine cervical human papillomavirus infection: a meta-analysis", BMC Infect Dis, 2011; 11: 10, doi:10.1186 / 1471-2334-11-10, https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC3023697 / .

[0253] 5.F.M.T. Lewis et al., "Vaginal Microbiome and Its Relationship to Behavior, Sexual Health, and Sexually Transmitted Diseases", Obstet Gynecol, author manuscript; available in PMC on September 13, 2019, doi:10.1097 / AOG.0000000000001932.

[0254] 6.K. Peebles et al., "High Global Burden and Costs of Bacterial Vaginosis: A Systematic Review and Meta-Analysis", Sex Transm Dis, May 2019; 46(5): 304 - 311, doi:10.1097 / OLQ.0000000000000972.

[0255] 7.J.D., "Sobel Bacterial vaginosis", Annu Rev Med, 2000; 51: 349 - 56. doi:10.1146 / annurev.med.51.1.349.

[0256] 8.M. Petrova et al., "Lactobacillus species as biomarkers and agents that can promote various aspects of vaginal health", Front Physiol. 2015; 6: 81, doi:10.3389 / fphys.2015.00081.

[0257] 9.J. Kaewsrichan et al., "Selection and identification of anaerobic lactobacilli producing inhibitory compounds against vaginal pathogens", FEMS Immunol Med Microbiol, October 2006; 48(1): 75 - 83, doi:10.1111 / j.1574 - 695X.2006.00124.x.

[0258] 10. J. Garza et al., "Cytokine profiles and Lactobacillus species presence in pre-menopausal subjects with genital Mycoplasma genitalium or Ureaplasma urealyticum colonization", Womens Health (Lond), 2021; 17: 17455065211009181, doi:10.1177 / 17455065211009181.

[0259] 11. W. Mendling., "Vaginal Microbiota", Adv Exp Med Biol, 2016; 902: 83 - 93, doi:10.1007 / 978-3-319-31248-4_6.

[0260] 12. A. Kiecka, "Can Lactobacillus spp. be a Factor Reducing the Risk of Miscarriage?", Pol J Microbiol., December 2021; 70(4): 431–446; Eckert LO et al., "Relationship of vaginal bacteria and inflammation with conception and early pregnancy loss following in vitro fertilization", Infect Dis Obstet Gynecol, (2003) 11: 11–7.

[0261] 13. Amabebe E, Anumba DOC, "The Vaginal Microenvironment: The Physiologic Role of Lactobacilli", Front Med (Lausanne), June 13, 2018; 5: 181.

[0262] 14. D. Eschenbach et al., "Influence of the Normal Menstrual Cycle on Vaginal Tissue, Discharge, and Microflora", Clinical Infectious Diseases, Vol. 30, No. 6, June 2000, pp. 901 - 907.

[0263] 15. S. Srinivasan et al., "Temporal Variability of Human Vaginal Bacteria and Relationship with Bacterial Vaginosis", PLoS One, 2010; 5(4): e10197.

[0264] 16. C. Bradshaw, J. D. Sobel, "Current Treatment of Bacterial Vaginosis - Limitations and Need for Innovation", J Infect Dis, August 15, 2016; 214(Suppl 1): S14–S20, doi:10.1093 / infdis / jiw159.

[0265] 17. J. Sobel, "Recurrent vulvovaginal candidiasis", Am J Obstet Gynecol, January 2016; 214(1): 15 - 21, doi:10.1016 / j.ajog.2015.06.067, Epub July 9, 2015.

[0266] 18. Vostral, SL, 2011, "Rely and Toxic Shock Syndrome: A Technological Health Crisis", Yale J. Biol. Med., 84, 447 - 459.

[0267] 19. Nicole, W., 2014, "A Question for Women's Health: Chemicals in Feminine Lubricants", Environmental Health Perspect 122, A70 - 75.

[0268] 20. Nonfoux L et al., "Impact of Currently Marketed Tampons and Menstrual Cups on Staphylococcus aureus Growth and Toxic Shock Syndrome Toxin 1 Production In Vitro", Appl Environ Microbiol, May 31, 2018; 84(12): e00351-18.

[0269] 21. S. Srinivasan et al., "Temporal Variability of Human Vaginal Bacteria and Relationship with Bacterial Vaginosis", PLoS One, 2010; 5(4): e10197.

[0270] 22. Janulaitiene M, Gegzna V, Baranauskiene L, A, Simanavicius M, Pleckaityte M, "Phenotypic characterization of Gardnerella vaginalis subgroups suggests differences in their virulence potential", PLoS One, July 12, 2018; 13(7): e0200625, doi:10.1371 / journal.pone.0200625.eCollection 2018.

[0271] 23. J. Sobel et al., "Conventional oral and secondary high dose vaginal metroni-dazole therapy for recurrent bacterial vaginosis: clinical outcomes, impacts of sex and menses", Infect Drug Resist, 2019; 12: 2297-2307, published online on July 24, 2019.

Claims

1. A sponge for vaginal use, comprising: a sponge body, wherein the sponge body has a porous structure, and the porous sponge structure forms at least a part of the outer surface of the sponge, Wherein, the porous sponge structure is made of a polysaccharide-based material containing glucomannan, wherein the sponge does not contain bacteria, and Wherein, the sponge body is impregnated with a liquid composition comprising one or more bacteriocins.

2. The sponge according to claim 1, wherein The liquid composition comprises a fixed amount of the one or more bacteriocins.

3. The sponge according to claim 1 or 2, wherein: The sponge was moistened.

4. A sponge according to any one of the preceding claims, wherein The polysaccharide-based material is glucomannan.

5. A sponge according to any one of the preceding claims, wherein The sponge is a cotton strip.

6. A sponge according to any one of the preceding claims, wherein The sponge body impregnated with the composition promotes vaginal microflora during menstruation, during the follicular phase, during the luteal phase, and / or during pregnancy.

7. A sponge according to any one of the preceding claims, wherein The sponge body impregnated with the composition promotes vaginal microbiota by inhibiting the growth of pathogens, particularly Staphylococcus aureus, Candida species and / or Gardnerella vaginalis, more particularly, wherein the growth of one or more pathogens is inhibited at least 10 times more than the growth of Lactobacillus gasseri and / or Lactobacillus crispatus over a period of 8 hours.

8. The sponge according to claim 7, wherein The porous sponge structure impregnated with the composition inhibits the growth of Gardnerella vaginalis compared to culture medium alone under the same conditions, wherein the growth is inhibited, for example, by at least 50 times, at least 100 times, or at least 250 times, or at least 500 times, or at least 1000 times, or at least 5000 times, or at least 10000 times, or at least 20000 times as tested by the tampon pouch method after 24 hours.

9. A sponge according to any one of the preceding claims, wherein When tested by the tampon pouch method after 8 h, the sponge body impregnated with the composition inhibited the growth of Lactobacillus up to 12 times compared to the culture medium alone under the same conditions.

10. A sponge according to any one of the preceding claims, wherein The liquid composition comprises an effective amount of one or more bacteriocins, in particular, wherein the effective amount inhibits the growth of Gardnerella vaginalis, Staphylococcus aureus and / or Candida species.

11. A sponge according to any one of the preceding claims, wherein The one or more bacteriocins are selected from class I bacteriocins, class II bacteriocins, class II bacteriocins and class IV bacteriocins from gram-positive bacteria, wherein the one or more bacteriocins are, for example, selected from class I bacteriocins and / or class II bacteriocins from gram-positive bacteria.

12. A sponge according to any one of the preceding claims, wherein The liquid composition comprises a bacteriocin.

13. A sponge according to any one of the preceding claims, wherein The one or more bacteriocins include nisin.

14. The sponge according to claim 13, wherein The nisin is nisin A and / or nisin Z.

15. A sponge according to any one of the preceding claims, wherein The liquid composition comprises a bacteriocin, which is nisin, in particular nisin A or nisin Z.

16. A sponge according to any preceding claim, wherein The composition further comprises lactic acid, wherein the composition comprises lactic acid, for example, in an amount that ensures that the pH value of the composition is from 3.5 to 5.5, or from 3.7 to 5, or from 3.9 to 4.5, or about 4.

2.

17. A method for making a sponge, the method comprising the steps of: providing a sponge body having a porous sponge structure, wherein the porous sponge structure forms at least a portion of an outer surface of the sponge; and The porous sponge structure is impregnated with a liquid composition comprising one or more bacteriocins to obtain a sponge according to any one of claims 1 to 16.

18. A liquid composition suitable for vaginal administration, comprising one or more bacteriocins in an amount that promotes vaginal microbiota by inhibiting the growth of one or more pathogens, in, The pH value of the composition is from 3.5 to 5.5, wherein the composition does not contain bacteria, and Wherein, the one or more bacteriocins include nisin.

19. The liquid composition according to claim 18, wherein The composition is an impregnation agent for a porous sponge.

20. The liquid composition according to claim 18 or 19, wherein The composition comprises a fixed amount of the one or more bacteriocins.

21. The liquid composition according to any one of claims 18 to 20, wherein The liquid composition comprises a bacteriocin.

22. The liquid composition according to claim 21, wherein The one bacteriocin is nisin, in particular nisin A or nisin Z.

23. The liquid composition according to any one of claims 18 to 22, wherein The one or more pathogens are selected from Staphylococcus aureus, Candida species, Gardnerella vaginalis, and combinations thereof.

24. The liquid composition according to any one of claims 18 to 23, wherein The total concentration of the one or more bacteriocins in the liquid composition is from 0.0001% (v / v) to 0.005% (v / v), from 0.0002% (v / v) to 0.002% (v / v), from 0.0004% (v / v) to 0.001% (v / v), from 0.0005% (v / v) to 0.0014% (v / v), from 0.0001% (v / v) to 0.0005% (v / v), 0.0014% (v / v) to 0.008% (v / v), in particular, wherein the total concentration of the one or more bacteriocins in the liquid composition is from 0.0006% (v / v) to 0.0013%, from 0.0007% (v / v) to 0.0012%, from 0.0008% (v / v) to 0.0011%, or from 0.0009% (v / v) to 0.0010%.

25. The liquid composition according to any one of claims 18 to 24, wherein The liquid composition is a gel, a solution or a lubricant, in particular, wherein the composition is a gel.

26. The liquid composition according to any one of claims 18 to 25, wherein The liquid composition is a gel, and wherein the gel further comprises glucomannan.

27. The liquid composition according to any one of claims 18 to 26, wherein The composition further comprises lactic acid, wherein the composition comprises lactic acid, for example, in an amount that ensures that the pH value of the composition is from 3.5 to 5.5, or from 3.7 to 5, or from 3.9 to 4.5, or about 4.

2.

28. The liquid composition according to any one of claims 18 to 26, wherein The composition inhibits the growth of one or more pathogens during menstruation, during the follicular phase, during the luteal phase, and / or during pregnancy.

29. A sponge according to any one of claims 1 to 16 or a liquid composition according to any one of claims 18 to 27 for use in inhibiting the growth of one or more pathogens associated with a condition or disease of the vagina.

30. A sponge according to any one of claims 1 to 16 or a liquid composition according to any one of claims 18 to 27 for use in preventing or treating dysbiosis.

31. A sponge according to any one of claims 1 to 16 or a liquid composition according to any one of claims 18 to 27 for use in preventing or treating vulvovaginal candidiasis and / or bacterial vaginosis.

32. A sponge according to any one of claims 1 to 16 or a liquid composition according to any one of claims 18 to 27 for use in preventing toxic shock syndrome.

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