Methods and compositions related to prebiotic formulations useful for promoting health of urogenital system

Abstract

The prebiotic formulation is capable of modulating the growth and / or activity of microorganisms in a region of the urogenital system. Such prebiotic formulations can comprise isomaltulose and at least one other prebiotic, such as 1-kestose or lactulose. These modulated microorganisms can be directly related to the health of the urogenital system, and their modulation can be used to prevent, control or treat dysbacteriosis in the region of the urogenital system.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 393,341, filed on July 29, 2022, which is incorporated herein by reference in its entirety. Background Art

[0003] Prebiotics are a group of nutrients that can be degraded by beneficial microorganisms present in the human body. The relationship between these microorganisms and overall health has become an area of ​​growing interest in recent years. Prebiotics are most commonly plant fibers that are selectively used as a food source by healthy bacteria to support their growth.

[0004] The term "microbiome" refers to a population of microorganisms that inhabit a specific biological niche. There are many such specific niches in the human body, for example, the intestine, skin, nasopharynx, mouth, and urogenital areas. The vaginal and urine microbiota represent important microbial niches. The urogenital microbiota can form a mutually beneficial relationship with its host, which has a significant impact on health and disease. Lactobacillus constitutes the majority of bacteria in the vagina and includes Lactobacillus crispatus ("L. crispatus"), Lactobacillus gasseri, Lactobacillus iners, and Lactobacillus jensenii (Verhelst et al., 2005; Chee et al., 2020). Maintaining a proper balance between bacterial species and ensuring the vigorous growth of beneficial bacteria (e.g., Lactobacillus crispatus) is critical to urogenital health.

[0005] To maintain proper urogenital health, it is important to both promote the growth of beneficial microorganisms and reduce the growth of undesirable microorganisms. Uropathogenic Escherichia coli (UPEC), Proteus mirabilis, Klebsiella pneumoniae, Staphylococcus saprophyticus, Enterococcus faecalis, and Streptococcus anginosus ("S. anginosus") are some of the bacteria that cause imbalance and infection in the urogenital area. Controlling the undesirable microbial population can help control or prevent infection, or alleviate symptoms such as urinary tract infection (UTI), urge urinary incontinence (UUI), and overactive bladder (OAB).

[0006] Isomaltulose (also referred to herein as Palatinose TM Isomaltulose (expressed as isomaltulose) is a prebiotic naturally present in products such as honey and sugar cane juice. Isomaltulose has been shown to promote the proliferation of specific probiotics (Su et al., 2021). In addition, the use of isomaltulose as a prebiotic in vaginal administration has been discussed in the art (U.S. Publication 2016 / 0375045A1 and U.S. Publication 2019 / 0231748A1). However, there is a continuing need in the art for prebiotic formulations that provide enhanced support for urogenital health by promoting the growth of beneficial microorganisms (e.g., Lactobacillus) while reducing the growth of undesirable microorganisms (e.g., Streptococcus anginosus). Summary of the invention

[0007] The present invention discloses a prebiotic preparation, which comprises the prebiotic agent isomaltulose and at least one of 1-kestose and lactulose. The combination of isomaltulose and at least one additional prebiotic (1-kestose or lactulose) produces an unexpected synergistic reaction, namely, it can increase the growth or activity of beneficial microorganisms (e.g., Lactobacillus crispatus) and reduce the activity growth of harmful or undesirable bacteria (e.g., Streptococcus anginosus).

[0008] Also disclosed is a method of modulating the growth of urogenital microorganisms in a subject, the method comprising administering a prebiotic formulation to the subject, wherein the prebiotic formulation comprises a prebiotic agent, and wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestulose and lactulose.

[0009] Also disclosed is a method for treating or preventing urogenital dysbacteriosis, the method comprising administering a prebiotic preparation to the subject, wherein the prebiotic preparation comprises a prebiotic agent, and wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestose and lactulose.

[0010] Additional aspects and advantages of the present disclosure will be set forth in part in the detailed description and any claims that follow, and will be derived in part from the detailed description, or may be learned by practicing the various aspects of the present disclosure. The advantages described below will be realized and obtained by means of the elements and combinations specifically indicated in the appended claims. It should be understood that the foregoing general description and the following detailed description both describe examples, are merely illustrative, and do not limit the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate certain examples of the present disclosure and, together with the description, serve to explain the principles of the present disclosure without limitation. In all drawings, like numerals represent like elements.

[0012] Figure 1 Graph showing mean log CFU / mL change (Δ) in bacterial recovery of Lactobacillus crispatus KC18-1173-1 and Streptococcus anginosus KC18-1131-3B after exposure to prebiotics in competition assays. Error bars represent standard error of the mean. Statistically significant differences are indicated by (*) (p<0.05).

[0013] Figure 2 Graph showing the mean log CFU / mL change (Δ) in bacterial recovery of Lactobacillus crispatus KC18-1173-1 and Streptococcus anginosus KC18-1131-3B after exposure to various combinations of isomaltulose and 1-kestose or isomaltulose and lactulose in competition assays. Error bars represent standard error of the mean. Statistically significant differences are indicated by (*) (p<0.05). DETAILED DESCRIPTION

[0014] definition

[0015] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, "a compound" includes a mixture of multiple compounds.

[0016] As used herein, the term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined (i.e., the limitations of the measurement system). For example, according to the practice in the art, "about" can mean within 3 standard deviations or more than 3 standard deviations. Alternatively, "about" can mean a range of up to 20% of a given value, or up to 10% of a given value, or up to 5% of a given value, or up to 1% of a given value. Alternatively, particularly for biological systems or processes, the term can mean within an order of magnitude, or within 5-fold, or within 2-fold of a value.

[0017] As used herein, the terms "inhibit," "inhibiting," and "inhibition" mean to reduce by a measurable amount or to completely prevent (eg, reduce an activity, response, condition, disease, or other biological parameter).

[0018] As used herein, "w / v" (or "wt / vol%" or "wt / vol") refers to the weight of a substance (in grams) divided by the volume of a solution (in milliliters) and then

[0019] Multiply the resulting value by 100.

[0020] As used herein, "urogenital system" refers to the vulva, vagina, urinary tract, bladder, and surrounding areas.

[0021] As used herein, the term "therapeutic effect" refers to the ability of the compositions and formulations of the present disclosure to stimulate the growth of beneficial microorganisms (e.g., Lactobacillus crispatus) relative to undesirable microorganisms (e.g., Streptococcus anginosus), as measured according to the therapeutic effect protocol described below. The therapeutic effect can be expressed as a ratio of beneficial microorganisms to undesirable microorganisms.

[0022] As used herein, "prebiotics" refers to ingredients that allow specific changes in both the composition and / or activity of the urogenital microbiota, which may (or may not) bring beneficial effects to the host subject. For example, in some cases, prebiotics can be added to materials worn by the host subject, such as diapers, pads, or liners. In other cases, prebiotics can be applied to the urogenital area in the form of creams, gels, sprays, or suppositories. Prebiotics can also be applied in the form of wipes. In addition to the combinations of prebiotics disclosed herein, prebiotics can also contain additional complex carbohydrates, amino acids, peptides, minerals, or other nutritional components that are essential for the survival of the bacterial composition.

[0023] "Colonization" of a host organism includes the non-temporary residence of bacteria or other microorganisms. As used herein, "reducing" the colonization of pathogenic or non-pathogenic bacteria in the urogenital tract (or any other microbial niche) of a host subject includes reducing the residence time of bacteria in the urogenital tract and reducing the number (or concentration) of bacteria in the urogenital tract. The reduction in colonization can be permanent or occur over a short period of time.

[0024] A "combination" of two or more bacteria includes the physical co-presence of the two bacteria, whether in the same material or product or in physically connected products, as well as the temporal co-administration or co-localization of the two bacteria.

[0025] As used herein, "treatment," "treat," or "treating" means a method of reducing the effects of a disease or condition. Treatment may also refer to a method of reducing the disease or condition itself, rather than just alleviating symptoms. Treatment may be any reduction relative to pre-treatment levels and may be, but is not limited to, complete elimination of a disease, condition, or symptom of a disease or condition.

[0026] As used herein, "dysbiosis" refers to a state of the microbiota or microbiome of the urogenital tract (or any other microbiota niche) in which the normal diversity and / or function of the ecological network is disrupted. Any disruption of the preferred (e.g., ideal) state of the microbiota can be considered dysbiosis, even if such dysbiosis does not result in a detectable decrease in health. Such a dysbiosis state may be unhealthy (e.g., leading to a disease state), or may only be unhealthy under certain conditions, or may prevent the subject from becoming healthier. Dysbiosis may be due to a reduction in the diversity of the composition of the microbial population, an overgrowth of one or more pathogenic populations (e.g., pathogenic populations) or pathogenic commensal populations, the presence and / or overgrowth of commensal organisms that can cause disease only when certain genetic and / or environmental conditions are present in the patient, or a shift to an ecological network that no longer provides a beneficial function to the host and therefore no longer promotes health.

[0027] As used herein, "microbiota" refers to the community of microorganisms that inhabit (persistently or transiently) a particular biological niche, such as a particular area of ​​a host subject (e.g., a mammal such as a human), including but not limited to eukaryotes (e.g., protozoa), archaea, bacteria, and viruses (including bacterial viruses, i.e., bacteriophages).

[0028] As used herein, "microbiome" refers to the genetic content of the community of microorganisms (including eukaryotes, archaea, bacteria, and viruses (including bacterial viruses (i.e., bacteriophages)) that live continuously or transiently in and on a specific ecological niche of a host subject, where "genetic content" includes genomic DNA, RNA (such as ribosomal RNA), epigenome, plasmids, and all other types of genetic information.

[0029] As used herein, "ecological niche" or simply "niche" refers to the ecological space occupied by an organism or group of organisms (e.g., a bacterial group). The niche describes how an organism (or group of organisms) responds to resource distribution, physical parameters (e.g., host tissue space), and competitors (e.g., growing when resources are abundant and / or when predators, parasites, and pathogens are scarce) and how it in turn changes these same factors (e.g., by limiting other organisms' access to resources or serving as a food source for predators).

[0030] As used herein, "preventing" or "prevention" refers to any method in which a disease state does not occur as a result of the action of a method (such as, for example, the administration of probiotics and / or prebiotics as described herein). In one aspect, it is understood that prevention can also mean that the disease does not develop to the extent that it occurs in an untreated control. For example, the frequency of disease occurrence can be reduced by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% relative to an untreated control. Thus, preventing a disease encompasses reducing the likelihood that a subject will develop a disease relative to an untreated subject (e.g., a subject not receiving a prebiotic as described herein).

[0031] As used herein, the term "subject" or "host subject" refers to any biological or animal subject (e.g., human) that is the object of a method or material. Synonyms used herein include "patient" and "host". In some embodiments, the subject or host may suffer from dysbacteriosis in the urogenital area. Examples include, but are not limited to, urinary tract infection (UTI) and urge urinary incontinence (UUI), common forms of urinary incontinence (UI), and overactive bladder (OAB).

[0032] As used herein, "synergy" or "synergy" refers to the interaction or cooperation of two or more entities to produce a combined effect that is greater than the sum of their individual effects. For example, "synergy" between two or more prebiotics can inhibit the growth ability of one or more pathogens, or can increase the growth ability of beneficial microorganisms.

[0033] General Description

[0034] Disclosed herein are prebiotic formulations that can modulate the growth and / or activity of microorganisms in the urogenital region. These modulated microorganisms can be directly related to urogenital health, and their modulation can be used to prevent, control or treat dysbacteriosis in the urogenital region. The female urogenital system has two major divisions, the urinary tract and the reproductive tract. The urinary tract consists of the kidneys, ureters, bladder and urethra. The reproductive tract consists of the clitoris, vagina, ovaries and uterus.

[0035] The prebiotic formulation disclosed herein comprises isomaltulose and at least one other prebiotic selected from 1-kestose and lactulose. Notably, the combination of isomaltulose and at least one additional prebiotic (1-kestose or lactulose) produces an unexpected synergistic reaction, i.e., it is capable of increasing the growth or activity of beneficial microorganisms and reducing the growth of harmful or undesirable bacterial activity.

[0036] The urogenital microbiota that can be regulated by the prebiotic formulations disclosed herein include, but are not limited to, Lactobacillus and Lactococcus. Lactobacillus and Lactococcus are both lactic acid bacteria, which are known for their ability to produce lactic acid as the sole or main end product of carbohydrate metabolism. They can prevent the adhesion, growth and colonization of pathogenic bacteria in the urinary tract. Therefore, increasing the population or activity of these microorganisms can be beneficial to the host and can be used to treat or prevent dysbiosis that may lead to certain infections, diseases or conditions. For example, studies have shown that healthy Lactobacillus microbiota has a strong inhibitory effect on the potential pathogenic bacteria Escherichia coli (Akgül et al., 2018). Examples of beneficial lactic acid producing bacteria (Lactobacillus and Lactococci) of the urogenital tract include, but are not limited to, Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus johnsonii, Lactobacillus mucosae, Lactobacillus mulieris, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus spp. rhamnosus), Lactobacillus reuteri, and Lactococcus lactis. In a specific embodiment, the growth of Lactobacillus crispatus can be promoted in the presence of a prebiotic formulation as described herein.

[0037] By "increase in the beneficial bacterial population" is meant an increase in the beneficial bacterial population by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0038] Prebiotic formulations disclosed herein can also be used to reduce the growth or activity of undesirable, destructive or harmful microorganisms in the urogenital system area. Examples of such bacteria include Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, saprophytic Staphylococcus, Enterococcus faecalis, Streptococcus anginosus, Candida albicans, Bacteroides fragilis, Peptostreptococcus anaerobius, Prevotella bivia, Atopobium vaginae, Mobiluncus mulieris, and Gardnerella vaginalis.

[0039] By "reduction in harmful or undesirable bacterial populations" is meant a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, %, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 91%, 99%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 91%, 92%, 94%, 96%, 99%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, 99%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99%, 91%, 92%, 93%, 94%, 0%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.

[0040] The inhibition of pathogen or non-pathogen encompasses the inhibition of any desired function or activity of pathogen or non-pathogen. Provided herein and those of ordinary skill in the art can recognize the display of inhibition, such as the reduction of pathogenic bacterial cell group growth or the reduction of pathogenic bacterial species colonization level. The inhibition of "growth" of pathogenic or non-pathogenic bacterial colonies can include the increase of inhibition of pathogenic or non-pathogenic bacterial cell group size and / or the proliferation (or breeding) of inhibition of pathogenic or non-pathogenic bacterial cell groups. The inhibition of pathogenic or non-pathogenic bacterial species colonization can be demonstrated by measuring and comparing the amount or load of bacterial species before and after treatment. As used herein, inhibition includes cell growth inhibition activity and / or cytotoxic activity.

[0041] Prebiotic composition

[0042] The prebiotic formulation disclosed herein comprises isomaltulose or its derivative as the first component. Isomaltulose is sold under the trade name Palatinose TMSale. Isomaltulose (6-0-α-D-pyranoglucopyranosyl-D-fructose) is a disaccharide carbohydrate composed of glucose and fructose. Glucose and fructose are linked by an α-1,6-glycosidic bond. Derivatives of isomaltulose are known to those skilled in the art and are contemplated herein (Weysser 2022; Barea-Alvarez 2014; Kashimura 2008, which are incorporated herein by reference for their teachings on derivatives of isomaltulose). Examples of derivatives include isomaltooligosaccharides, which are glucose oligomers with α-D-(1,6)-bonds (examples are isomaltose, panose, isomaltotriose, isomaltotetraose and isomaltopentose). These isomaltooligosaccharides may contain 3, 4, 5, 6, 7, 8 or 9 monosaccharide units. Although isomaltulose has previously been found to be useful for modulating the microbiota of the urogenital region (U.S. Patent Application Publications 2016 / 0375045A1 and 2019 / 0231748A1, both of which are incorporated herein by reference in their entirety for their teachings on the use of isomaltulose to treat urogenital conditions), it has now been surprisingly and unexpectedly discovered that the combination of isomaltulose with 1-kestose or lactulose or their derivatives provides a synergistic effect in modulating the urogenital microbiota. The synergistic combination of these prebiotics is described in Example 1 and can be found at Figure 1 middle.

[0043] 1-Kestose is a trisaccharide found in vegetables and is composed of β-D-fructofuranosyl and α-D-glucopyranosyl residues attached at positions 1 and 2, respectively. Methods for obtaining 1-kestose are known to those skilled in the art (US20150232898A1, which is incorporated herein by reference for its teachings on obtaining 1-kestose). Lactulose is a synthetic galactosyl fructose disaccharide (e.g., by Also disclosed are derivatives of lactulose.

[0044] By "at least one of" 1-kestose and lactulose is meant that at least one of these compounds (1-kestose or lactulose) is present together with isomaltulose. Thus, for example, a prebiotic preparation may comprise (i) isomaltulose and 1-kestose or (ii) isomaltulose and lactulose.

[0045] The prebiotic formulations disclosed herein may include a carrier. The carrier may be physiologically compatible with the dermal or epithelial tissue of the subject to which it is administered. For example, the carrier may be a "dermatologically acceptable carrier," which refers to a carrier suitable for topical application to the urogenital region. That is, the carrier is preferably substantially inactive except for the surfactant properties used to prepare a suspension of the active ingredient. The composition may include other physiologically active ingredients that do not interfere with the efficacy of the active agent in the composition.

[0046] Dermatologically acceptable carriers can be in a variety of forms, such as simple solutions (water-based or oil-based), solid forms (e.g., gels or sticks), and emulsions. In some embodiments, the carrier can be aqueous or non-aqueous. Water is a particularly preferred aqueous carrier. Non-aqueous carriers can include, for example, glycols such as propylene glycol, butylene glycol, triethylene glycol, hexylene glycol, polyethylene glycol, ethoxydiglycol, and dipropylene glycol; alcohols such as ethanol, n-propanol, and isopropanol; triglycerides; ethyl acetate; acetone; triacetin; and combinations thereof.

[0047] The carrier may comprise more than 75% w / v of the prebiotic formulation. For example, the carrier may comprise 75% w / v, 76% w / v, 77% w / v, 78% w / v, 79% w / v, 80% w / v, 81% w / v, 82% w / v, 83% w / v, 84% w / v, 85% w / v, 86% w / v, 87% w / v, 88% w / v, 89% w / v, 90% w / v, 91% w / v, 92% w / v, 93% w / v, 94% w / v, 95% w / v, 96% w / v, 97% w / v, 98% w / v, or 99% w / v of the pharmaceutical composition, or any amount greater than, less than, or between these values.

[0048] Prebiotic compounds are considered the "active portion" of a prebiotic preparation. The total weight of prebiotic compounds (meaning, as a whole, isomaltulose and at least one other prebiotic compound) in a prebiotic preparation may be 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.50%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.59%, 0.60%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.70%, 0.71%, 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.59%, 0.6%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.7%, 0.71%, 0.72%, 0.73%, 0.74%, 0. 75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.8%, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0.88%, 0.89%, 0.9%, 0.91%, 0.92%, 0.93%, 0.94%, 0.95%, 0.96%, 0.97%, 0.98%, 0.99%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9 %, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8％、7.9％、8.0％、8.1％、8.2％、8.3％、8.4％、8.5％、8.6％、8.7％、8.8％、8.9％、9.0％、9.1％、9.2％、9.3％、9.4％、9.5％、9.6％、9.7％、9.8％、9.9％、10.0％、10.1％、10.2％、10.3％、10.4％、10.5％、10.6％、10.7％、10.8％、10.9％、11.0％、11.1％、11.2％、11.3％、11.4％、11.5％、11.6％、11.7％、11.8％、11.9％、12.0％、12.1％、12.2％、12.3％、12.4％、12.5％、12.6％、12.7％、12.8％、12.9％、13.0％、13.1％、13.2％、13.3％、13.4％、13.5％、13.6％、13.7％、13.8％、13.9％、14.0％、14.1％、14.2％、14.3％、14.4％、14.5％、14.6％、14.7％、14.8％、14.9％、15.0％、15.1％、15.2％、15.3％、15.4％、15.5％、15.6％、15.7％、15.8％、15.9％、16.0％、16.1％、16.2％、16.3％、16.4％、16.5％、16.6％、16.7％、16.8％、16.9％、17.0％、17.1％、17.2％、17.3％、17.4％、17.5％、17.6％、17.7％、17.8％、17.9％、18.0％、18.1％、18.2％、18.3％、18.4％、18.5％、18.6％、18.7％、18.8％、18.9％、19.0％、19.1％、19.2％、19.3％、19.4％、19.5％、19.6％、19.7％、19.8％、19.9％、20.0％、20.1％、20.2％、20.3％、20.4％、20.5％、20.6％、20.7％、20.8％、20.9％、21.0％、21.1％、21.2％、21.3％、21.4％、21.5％、21.6％、21.7％、21.8％、21.9％、22.0％、22.1％、22.2％、22.3％、22.4％、22.5％、22.6％、22.7％、22.8％、22.9％、23.0％、23.1％、23.2％、23.3％、23.4％、23.5％、23.6％、23.7％、23.8％、23.9％、24.0％、24.1％、24.2％、24.3％、24.4％、24.5％、24.6％、24.7％、24.8%, 24.9% or 25.0%, or any amount less than, greater than, or between these values. In a preferred embodiment, the total prebiotic compound comprises 0.5 w / v to 10% w / v. In another preferred embodiment, the prebiotic compound comprises 0.5% w / v to 5% w / v. In yet another preferred embodiment, the prebiotic compound comprises 2% w / v. .

[0049] Specifically, isomaltulose can be present in the prebiotic preparation in the following amounts: 0.1% w / v, 0.2% w / v, 0.3% w / v, 0.4% w / v, 0.5% w / v, 0.6% w / v, 0.7% w / v, 0.8% w / v, 0.9% w / v, 1.0% w / v, 1.1% w / v, 1.2% w / v, 1.3% w / v, 1.4% w / v, 1.5% w / v, 1.6% w / v, 1.7% w / v, 1.8% w / v, 1.9% w / v, 2.0% w / v, 2.1% w / v, 2.2% w / v, 2.3% w / v, 2.4% w / v, 2.5% w / v, 2.6% w / v, 2.7% w / v, 2.8% w / v, 2.9% w / v, 3.0% w / v, 3.1% w / v, 3.2% w / v, 3.3% w / v, 3.4% w / v, 3.5% w / v, 3.6% w / v, 3.7% w / v, 3.8% w / v, 3.9% w / v, 4.0% w / v, 4.1% w / v, 4.2% w / v, 4.3% w / v, 4.4% w / v, 4.5% w / v, 4.6% w / v, 4.7% w / v, 4.8% w / v, 4.9% w / v, 5.0% w / v, 5.1% w / v, 5.2% w / v, 5.3% w / v, 5.4% w / v, 5.5% w / v, 5.6% w / v, 5.7% w / v, 5.8% w / v, 5.9% w / v, 6.0% w / v, 6.1% w / v, 6.2% w / v, 6.3% w / v, 6.4% w / v, 6.5% w / v, 6.6% w / v, 6.7% w / v, 6.8% w / v, 6.9% w / v, 7.0% w / v, 7.1% w / v, 7.2% w / v, 7.3% w / v, 7.4% w / v, 7.5% w / v, 7.6% w / v, 7.7% w / v, 7.8% w / v, 7.9% w / v, 8.0% w / v, 8.1% w / v, 8.2% w / v, 8.3% w / v, 8.4% w / v, 8.5% w / v, 8.6% w / v, 8.7% w / v, 8.8% w / v, 8.9% w / v, 9.0% w / v, 9.1% w / v, 9.2% w / v, 9.3% w / v, 9.4% w / v, 9.5% w / v, 9.6% w / v, 9.7% w / v, 9.8% w / v, 9.9% w / v, 10.0% w / v, 10.1% w / v, 10.2% w / v, 10.3% w / v, 10.4% w / v, 10.5% w / v, 10.6% w / v, 10.7% w / v, 10.8% w / v, 10.9% w / v, 11.0% w / v, 11.1% w / v, 11.2% w / v, 11.3% w / v, 11.4% w / v, 11.5% w / v, 11.6% w / v, 11.7% w / v, 11.8% w / v, 11.9% w / v, 12.0% w / v, 12.1% w / v, 12.2% w / v, 12.3% w / v, 12.4% w / v or 12.5% ​​w / v, or any amount greater than, less than, or between these values. .

[0050] The additional prebiotic compound of the preparation (1-kestotriose or lactulose, or their derivatives) may be present in the following amounts: 0.1% w / v, 0.2% w / v, 0.3% w / v, 0.4% w / v, 0.5% w / v, 0.6% w / v, 0.7% w / v, 0.8% w / v, 0.9% w / v, 1.0% w / v, 1.1% w / v, 1.2% w / v, 1.3% w / v, 1.4% w / v, 1.5% w / v, 1.6% w / v, 1.7% w / v, 1.8% w / v, 1.9% w / v, 2.0% w / v, 2.1% w / v, 2.2% w / v, 2.3% w / v, 2.4% w / v, 2.5% w / v, 2.6% w / v, 2.7% w / v, 2.8% w / v, 2.9% w / v, 3.0% w / v, 3.1% w / v, 3.2% w / v, 3.3% w / v, 3.4% w / v, 3.5% w / v, 3.6% w / v, 3.7% w / v, 3.8% w / v, 3.9% w / v, 4.0% w / v, 4.1% w / v, 4.2% w / v, 4.3% w / v, 4.4% w / v, 4.5% w / v, 4.6% w / v, 4.7% w / v, 4.8% w / v, 4.9% w / v, 5.0% w / v, 5.1% w / v, 5.2% w / v, 5.3% w / v, 5.4% w / v, 5.5% w / v, 5.6% w / v, 5.7% w / v, 5.8% w / v, 5.9% w / v, 6.0% w / v, 6.1% w / v, 6.2% w / v, 6.3% w / v, 6.4% w / v, 6.5% w / v, 6.6% w / v, 6.7% w / v, 6.8% w / v, 6.9% w / v, 7.0% w / v, 7.1% w / v, 7.2% w / v, 7.3% w / v, 7.4% w / v, 7.5% w / v, 7.6% w / v, 7.7% w / v, 7.8% w / v, 7.9% w / v, 8.0% w / v, 8.1% w / v, 8.2% w / v, 8.3% w / v, 8.4% w / v, 8.5% w / v, 8.6% w / v, 8.7% w / v, 8.8% w / v, 8.9% w / v, 9.0% w / v, 9.1% w / v, 9.2% w / v, 9.3% w / v, 9.4% w / v, 9.5% w / v, 9.6% w / v, 9.7% w / v, 9.8% w / v, 9.9% w / v, 10.0% w / v, 10.1% w / v, 10.2% w / v, 10.3% w / v, 10.4% w / v, 10.5% w / v, 10.6% w / v, 10.7% w / v, 10.8% w / v, 10.9% w / v, 11.0% w / v, 11.1% w / v, 11.2% w / v, 11.3% w / v, 11.4% w / v, 11.5% w / v, 11.6% w / v, 11.7% w / v, 11.8% w / v, 11.9% w / v, 12.0% w / v, 12.1% w / v, 12.2% w / v, 12.3% w / v, 12.4% w / v or 12.5% ​​w / v, or any amount greater than, less than, or between these values. .

[0051] For example, when the prebiotic formulation comprises isomaltulose and an additional prebiotic compound selected from the group consisting of 1-kestose and lactulose, isomaltulose may be present at 0.5% w / v and the additional prebiotic may be present at 1.5% w / v. In another embodiment, isomaltulose may be present at 0.7% w / v and the additional prebiotic compound may be present at 1.3% w / v. Alternatively, isomaltulose may be present at 0.8% w / v and the additional prebiotic compound may be present at 1.2% w / v. In another embodiment, isomaltulose may be present at 1.0% w / v and the additional prebiotic may be present at 1.5% w / v.

[0052] The compound may be present at 1.0% w / v. Alternatively, isomaltulose may be present at 1.2% w / v and the additional prebiotic compound may be present at 0.8% w / v. Alternatively, isomaltulose may be present at 1.3% w / v and the additional prebiotic compound may be present at 0.7% w / v. Alternatively, isomaltulose may be present at 1.5% w / v and the additional prebiotic compound may be present at 0.5% w / v. In another embodiment, isomaltulose may be present at 2.0% w / v and the additional prebiotic compound may be present at 1.0% w / v. Alternatively, isomaltulose may be present at 2.0% w / v and the additional prebiotic compound may be present at 2.0% w / v. In another embodiment, isomaltulose may be present at 2.0% w / v and the additional prebiotic compound may be present at 0.2% w / v. Alternatively, isomaltulose may be present at 2.0% w / v and the additional prebiotic compound may be present at 0.1% w / v. In another embodiment, when isomaltulose is present at 1.5% w / v, the additional prebiotic may be present at 1.5% w / v. In other words, the ratio of isomaltulose to the additional prebiotic compound may be 20:1, 13:7, 10:1, 5:1, 4:1, 3:1, 3:2, 2:1, 1:1, 1:2, 2:3, 1:3, 1:4, 1:5, 7:13, or any amount greater than, less than, or between these ratios.

[0053] Prebiotic preparations may additionally include probiotic compositions. It may include live, active cultures. Examples of such cultures are known to those skilled in the art and are found in U.S. Patent Application 20200138881A1, Japanese Patent Application JP6509822B2, PCT Application WO2022100633A1, and Chinese Patent Application CN110960561A, which are incorporated herein by reference for the teachings of probiotic compositions that can be topically applied to the urogenital system region. Specific examples include Lactobacillus crispatus (Li et al., 2019, the disclosure of which is incorporated herein by reference for the topical application of Lactobacillus crispatus), Lactobacillus gasseri, Lactobacillus fermentum, and Lactobacillus rhamnosus.

[0054] The prebiotic formulations disclosed herein may include known antimicrobial agents, known antiviral agents, known antifungal agents, all of which must be compatible with maintaining the viability of the prebiotic compounds. Other agents in the composition may be potentiators or active agents. The composition may additionally contain known antioxidants,

[0055] Buffering agents and other agents to improve appearance, smell or texture. Thickening agents such as polyvinyl pyrrolidone, polyethylene glycol or carboxymethyl cellulose may be added to the composition.

[0056] Other suitable additives that can be included in the microbial compositions of the present disclosure include compatible colorants, deodorants, emulsifiers, defoamers (when foam is not required), lubricants, skin conditioners, skin protectants and skin benefit agents (e.g., aloe and tocopheryl acetate), solvents (e.g., water-soluble ethylene glycols and ethylene glycol ethers, glycerol, water-soluble polyethylene glycols, water-soluble polyethylene glycol ethers, water-soluble polypropylene glycols, water-soluble polypropylene glycol ethers, dimethyl isosorbide), solubilizers, suspending agents, builders (e.g., alkali metal and alkaline earth metal salts of carbonates, bicarbonates, phosphates, hydrogen phosphates, dihydrogen phosphates, hydrogen sulfates), wetting agents, pH adjusting ingredients (suitable pH ranges for the compositions can be 3.5 to 8), chelating agents, propellants, dyes and / or pigments, and combinations thereof.

[0057] How to use prebiotic preparations

[0058] Disclosed herein are methods of modulating the growth of urogenital microorganisms in a subject, the method comprising administering a prebiotic formulation to the subject, wherein the prebiotic formulation comprises a prebiotic agent, and wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestose and lactulose. As described above, modulating can mean increasing the growth of desired microorganisms, and / or decreasing or reducing the growth of undesirable microorganisms.

[0059] Specifically, disclosed herein is a method for treating or preventing urogenital dysbacteriosis, the method comprising administering a prebiotic preparation to the subject, wherein the prebiotic preparation comprises a prebiotic agent, and wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestulose and lactulose, as described above.

[0060] Imbalanced bacterial flora may lead to a variety of diseases, conditions, infections and other uncomfortable or undesirable physical effects. Examples include, but are not limited to, overactive bladder, urge incontinence or urinary tract infections.

[0061] Prior to administration of the prebiotic composition, the subject may be diagnosed as having overactive bladder, urge incontinence, or a urinary tract infection. For example, a physician or other caregiver may provide such a diagnosis. In another example, the subject may be diagnosed as being susceptible to overactive bladder, urge incontinence, or a urinary tract infection. In another example, the subject may use the prebiotic composition as a preventative measure, or a way to prevent overactive bladder, urge incontinence, or a urinary tract infection. The methods and compositions disclosed herein may be used as a regular part of health or may be used as needed.

[0062] The prebiotic preparations disclosed herein can be topically applied. For example, the prebiotic preparation can be placed on or integrated into an absorbent product that contacts the urogenital region of the subject, such as a sanitary product (sanitary pad or tampon), a diaper, a paper towel, toilet paper, or a wipe. Other examples include skin patches, tapes, clothing, panty protectors, or incontinence pads. The prebiotic preparation can be placed on or in the product before the product reaches the end user so that the preparation is integrated into the product before the product reaches the subject, or it can be placed on the product immediately before the subject uses it. In this case, the preparation can be placed on a substrate in the form of a spray, a lotion, a cream, a soap, a liquid, or a gel. In either case, the use of an absorbent product coated with a prebiotic preparation can provide contact between the urogenital region and the absorbent product, thereby contacting the target area with an effective amount of the prebiotic preparation.

[0063] Compositions disclosed herein can be applied to suitable substrates, which can then be used to apply therapeutic agents to users. Suitable application devices include fiber webs, such as wet-laid tissue webs or air-laid fiber webs, gauze, cotton swabs, transdermal patches, containers or retainers. Particularly preferred application devices include fiber webs, including washable and non-washable cellulose webs and non-woven fiber webs of synthetic fiber materials. Available fiber webs can be wet-laid fiber webs, air-laid fiber webs, melt-blown fiber webs or spunbond fiber webs. Suitable synthetic fiber materials include melt-blown polyethylene, polypropylene, copolymers of polyethylene and polypropylene, bicomponent fibers comprising polyethylene or polypropylene, etc. Available non-woven fiber webs can be melt-blown fiber webs, spunbond fiber webs, spunbond fiber webs, air-laid fiber webs, spunlace entangled non-woven fiber webs, spunlace fiber webs or bonded combed fiber webs.

[0064] In certain embodiments, particularly those in which the composition is applied to a web, it may be desirable for the formulation to provide certain physical attributes, such as having a smooth, lubricious, non-greasy texture; being able to at least partially transfer from the web to a user; being able to remain on the web at room temperature; or being compatible with the web manufacturing process. In certain embodiments, it is preferred that at least a portion of the composition transfers from the tissue to the user during use.

[0065] The composition can be applied to the web during the process of forming the web or after the web has been formed and dried, the latter of which is often referred to as an off-line process or post-processing. Suitable methods for applying the composition to the web include methods known in the art, such as gravure printing, flexographic printing, spraying, WEKO TM , slot coating or electrostatic spraying. A particularly preferred off-line application method is rotogravure printing.

[0066] Products

[0067] Also disclosed herein are various articles of manufacture that combine the prebiotic formulations described herein with various medical or personal hygiene devices to modulate the urogenital microbiota. Thus, contemplated herein is a prebiotic formulation comprising isomaltulose and one other prebiotic, such as 1-kestose or lactulose, applied to a solid surface or impregnated into a solid matrix of any device or article intended to contact the urogenital region of a subject. Preferably, the solid surface is a flexible matrix that can be worn or wiped on the skin or mucous membranes. For example, the solid surface can be absorbent.

[0068] Many different types of absorbent products having absorbent structures are well known in the art and may include diapers, towelettes (e.g., baby wipes or feminine hygiene towelettes), sanitary napkins, tampons, panty protectors, skin patches, tapes, bandages, wound or sore dressings, absorbent pads, incontinence pads, bed sheets or protectors, saliva absorbents, articles of clothing (e.g., underwear, pajamas), bath towels, towels, toilet paper, paper towels, etc. Accordingly, described herein is an absorbent product comprising an aqueous liquid absorbent structure and an effective amount of a prebiotic formulation as described herein.

[0069] Hygiene products (e.g., absorbent products such as Absorbent structures in sanitary products are usually produced by loosening cellulose or other fiber pulp into rolls, bales or sheets, for example, to form pulp pads, sometimes with so-called superabsorbent materials mixed in the pulp pads. Superabsorbent materials are usually polymer preparations that are able to absorb several times their own weight in body fluids and are well known in the art. Pulp pads are usually compressed to increase their ability to wick liquids while also reducing the volume of the pulp body, thereby obtaining a product that is as compact as possible to achieve the absorbent properties required for a particular hygiene product.

[0070] The absorbent structure may additionally contain other ingredients, for example, components known in the art that improve liquid acquisition properties, liquid wicking properties, liquid retention properties, and the like. Other included ingredients include components that increase cohesive strength (i.e., the ability to withstand deformation during use). The absorbent structure may contain fibrous woven, knitted or nonwoven materials, occlusive or non-occlusive films or membranes, particles, granules or aggregates of absorbent materials, synthetic polymer fibers, films, membranes and foams (e.g., nylon, polytetrafluoroethylene (PTFE, such as or ), polystyrene, polycarbonate, polyvinyl chloride, and polysulfone). All of these forms are well known in the art and include, for example, knitted or woven fabrics, nonwoven fabrics such as felt and batting, fiber balls of cotton, rayon, cellulose or synthetic fibers, and similar materials.

[0071] The fiber can be a natural fiber, including but not limited to natural fibers such as fluff, silk, cotton, and cellulose fibers. Natural polymers based on polysaccharides can also be used, including but not limited to: modified cellulose and cellulose derivatives (e.g., alkyl cellulose, hydroxyalkyl cellulose, carboxymethyl cellulose); gum resins (e.g., guar gum, locust bean gum, tragacanth gum, gum arabic, pectin, etc.); starch and starch derivatives (e.g., corn starch, corn starch, potato starch, amylose, amylopectin, dextrin, dextran, modified starch, hydroxyethyl starch, cationic starch, starch graft polymers, and similar polymers). The fiber can be a synthetic fiber, including but not limited to: polyester, polyolefin, polyamide, polyvinyl alcohol, polyvinyl acetate, polyvinyl chloride, polyethylene urea, polyurethane, polyurea, polyacrylonitrile, and copolymers of these polymers.

[0072] Absorbent products can be formed in a multi-layer configuration format having an absorbent structure layer, a fluid-permeable top layer that allows liquid wicking but is inherently non-wettable due to its structural composition (e.g., synthetic fiber construction), and a fluid-impermeable bottom layer (i.e., backsheet) that prevents the absorbed fluid from being transferred from the absorbent structure layer to the adjacent tissues of the user when in contact with the absorbent product during use. Such layered configurations are well known in the diaper and panty liner arts.

[0073] The prebiotic formulation can be applied to a solid surface using any of a variety of known methods, including, for example, applying a powder, spray drying the prebiotic formulation onto a material, or soaking the material in a solution containing the prebiotic formulation and then using the wet material or drying the material before use. A porous material can contain the prebiotic compound in the pores or interstices of the solid material. The prebiotic formulation can be attached by adhesion, such as to an adhesive layer, which is then applied to the skin (e.g., a bandage or skin patch). The prebiotic formulation and / or the isolated active agent can be impregnated into the solid material during the manufacturing process of the flexible article (e.g., added to the synthetic composition before or during the polymerization process).

[0074] Any of the solid materials comprising the prebiotic formulation can be packaged individually or in groups suitable for containing materials treated using standard packaging materials (e.g., in shrink wrap, sealed bags, protective packaging materials, or dispensing containers suitable for containing dry or wet materials). The article can have applied thereto any of the additional / optional components of the composition of the present disclosure, including carriers, bactericides, antibacterial agents, salts, FOS, and the like. In particular, the absorbent product can include inert ingredients, neutral fillers, and the like as an integral part of the absorbent structure. Examples of neutral fillers include peat, sand, clay, garden molds, crushed shells of nuts or kernels, wood flour, chitin-containing flour, and similar well-known materials.

[0075] Unless otherwise defined, all scientific and technical terms used herein have the same meaning as those generally understood by those skilled in the relevant art. Unless otherwise stated, the techniques employed or considered herein are standard methods well known to those of ordinary skill in the art. These examples are for illustrative purposes only.

[0076] Example

[0077] To further demonstrate the principles of the present disclosure, the following examples are presented to provide a person of ordinary skill in the art with a complete disclosure and description of how to prepare and evaluate the compositions, articles and methods claimed herein. They are intended to serve as examples and are not intended to limit the scope of their disclosure as the inventors believe. Efforts have been made to ensure the accuracy of numbers (e.g., amounts, temperatures, etc.); however, some errors and deviations should be considered. Unless otherwise stated, temperatures are ° C or ambient temperature, and pressures are atmospheric or near atmospheric. There are many variations and combinations of process conditions that can be used to optimize product quality and performance. Such process conditions can be optimized by simply conducting reasonable, routine experiments.

[0078] Example 1: Screening of prebiotic combinations in co-culture assays

[0079] Prebiotic Palatinose TM / Isomaltulose for Lactobacillus crispatus In this example, prebiotics were combined with five additional compounds (1-kestose, D-lactitol, lactulose, maltitol, and α-methyl-D-glucoside). This was done to evaluate the effects of Palatinose when combined with secondary compounds. TM The effects of Palatinose (i.e., increasing the recovery rate of Lactobacillus crispatus in the co-culture system and reducing the recovery rate of Streptococcus anginosus in the co-culture system) can be synergistic or further increased. Table 1 lists the effects of Palatinose TM / isomaltulose prebiotic compound mixture. Five additional compounds were selected based on their positive or neutral Lactobacillus crispatus recoveries when tested against S. anginosus in a competition assay using the individual compounds. The compound mixtures are listed in Table 2.

[0080] method

[0081] The method used in this article is adapted from the method of Vongsa et al. The method used in this specific embodiment is as follows:

[0082] 1. Use single prebiotics, prebiotic mixtures or positive control (glucose) in API 50 CHL medium (Biomerieux, Marcy-l' The storage medium was produced in France. API 50 CHL medium contains a minimum amount of carbon for growth and bromocresol purple as a pH indicator for fermentation. The storage medium was filter sterilized with a 0.22 μM filter and stored at 4°C-6°C.

[0083] 2. From frozen stock culture, bacteria were subcultured twice into De Man, Rogosa, and Sharpe (MRS) broth (BD Difco, Becton Dickinson, Franklin Lakes, NJ) and incubated anaerobically overnight at 37°C until quiescent.

[0084] 3. Plate the second generation MRS broth subculture onto MRS agar plates (for Lactobacillus crispatus KC18-1173-1) or blood agar plates (for Streptococcus anginosus KC18-1131-3B) and incubate anaerobically at 37°C overnight until stationary.

[0085] 4. Use a sterile pointed swab to transfer each bacterial colony to API suspension medium (Biomerieux, Marcy-l' France) to achieve a turbidity twice that of the MacFarland standard, thereby producing a bacterial suspension.

[0086] 5. Count the starting culture.

[0087] 6. Add 9.8 mL of prebiotic medium to a sterile 15 mL conical tube.

[0088] 7. 100 μL of each prepared bacterial suspension was added to the culture medium and cultured anaerobically at 37°C for 48 hours.

[0089] 8. Harvest 100 μL from each tube containing bacteria and prebiotic media, make serial dilutions, and plate on MRS agar and tryptic soy agar (TSA).

[0090] 9. Count bacteria (both Lactobacillus crispatus and Streptococcus anginosus) on MRS agar. Small colony counts are Streptococcus anginosus and large colony counts are Lactobacillus crispatus.

[0091] 10. S. anginosus has been enumerated on TSA plates. Lactobacillus species do not grow well on TSA, whereas S. anginosus grows well.

[0092] 11. The background growth of bacteria in the negative control (CHL medium without prebiotics) was subtracted from the recovered growth in the presence of the prebiotic mixture.

[0093] 12. Statistical analysis was performed using Student t test, with the significance level set at p < 0.05.

[0094] result

[0095] The data generated here differ from previous findings with single prebiotic compounds because the secondary compounds are similar to Palatinose TM / isomaltulose. The recovery of Lactobacillus crispatus for some prebiotic mixtures was similar to that of the previous method using only Palatinose TM The observed findings were comparable ( Table 3 ; Figure 1 However, when minor compounds such as 1-kestose or lactulose were added to the mixture, the recovery of S. anginosus decreased ( Figure 2 ). Since other pathogenic bacteria use the compounds 1-kestose and lactulose, they are not used alone. The reduction in the recovery of S. anginosus may be due to the rapid metabolism of 1-kestose (trisaccharide) and lactulose (disaccharide) by Lactobacillus crispatus, resulting in the production of lactic acid that is released into the external environment. Lactic acid-producing bacteria, such as Lactobacillus crispatus, have antimicrobial effects against certain pathogenic bacterial species.

[0096] The data contained the unexpected finding of statistical significance (p<0.05) with the following prebiotic combinations showing the best activity against Lactobacillus crispatus when tested in a competition assay:

[0097] Lactulose (1.5% w / v) + Palatinose TM (0.5% w / v) and lactulose (1.3% w / v) + Palatinose TM (0.7%w / v)

[0098] ○Palatinose TM (2% w / v) itself supported a ratio of Lactobacillus crispatus to S. anginosus of 2,681.

[0099] o Lactulose (2% w / v) by itself supported a ratio of Lactobacillus crispatus to S. anginosus of 37,556.

[0100] ○ However, in the above combination, with Palatinose TM Compared with the control group (2.0% w / v), the ratios of Lactobacillus crispatus to S. anginosus showed statistical significance (p<0.05) at 932,446 and 572,164, respectively.

[0101] ■ In addition, the change in S. anginosus log CFU / mL after the competition assay was statistically significant (p<0.05).

[0102] ○ Two additional Lactobacillus crispatus to S. anginosus ratios with statistical significance (<0.05) were lactulose (1.0% w / v) + Palatinose TM (2.0% w / v) and lactulose (0.1% w / v) + Palatinose TM (2.0% w / v), 243,333 and 17,413 respectively.

[0103] ■However, for lactulose (1.0% w / v) + Palatinose TM (2.0% w / v), only the change in logCFU / mL of S. anginosus was statistically significant (p<0.05).

[0104] ● 1-Kestose ranging from 0.1% (w / v) to 1.5% (w / v) + Palatinose ranging from 0.5% (w / v) to 2.0% (w / v) TM Combination of

[0105] ○ Palatinose (2% w / v) by itself supported a ratio of Lactobacillus crispatus to S. anginosus of 2,681.

[0106] o The above combinations were statistically significant (p<0.05), producing ratios of L. crispatus to S. anginosus ranging from 13,005 to 322,274.

[0107] ■ In addition, most combinations (except 1-kestose (0.1% w / v) + Palatinose TM (2.0% w / v) and 1-kestose (0.2% w / v) + Palatinose TM (except 2.0% w / v)) showed that the changes in log CFU / mL of S. anginosus were statistically significant (p<0.05).

[0108] ○ Although 1-kestose (2.0% w / v) + Palatinose TM (2.0% w / v) and 1-kestose (1.0% w / v) + Palatinose TM (2.0% w / v) showed a ratio of Lactobacillus crispatus to S. anginosus of 389, 223 and 336, 363, respectively, but the result was not statistically significant (p>0.05).

[0109] In view of these results, the combination of isomaltulose and at least one additional prebiotic (1-kestose or lactulose) produces an unexpected synergistic response, namely being able to increase the growth or activity of beneficial microorganisms and reduce the growth of harmful or undesirable bacterial activity.

[0110] The combination of prebiotics can be used in a variety of product forms, including moistened and dry wipes, incorporated into pads, panty liners and absorbent garments, formulated into products including lotions, creams, gels, sprays and suppositories.

[0111] Finally, while the present disclosure has been presented in detail with respect to certain illustrative and specific aspects thereof, the present disclosure should not be construed as limited thereto since various modifications may be made without departing from the broader spirit and scope of the present disclosure as defined in the appended claims.

[0112] It will be apparent to those skilled in the art that various modifications and variations may be made to the present disclosure without departing from the scope or spirit of the present disclosure. Other embodiments of the present disclosure will be apparent to those skilled in the art by considering the practice of the specification and method disclosed herein. It is contemplated that the specification and embodiments are only considered as examples.

[0113] surface

[0114] Table 1. Prebiotic compounds tested.

[0115]

[0116] 1 Positive controls for growth and fermentation.

[0117] Table 2. Prebiotic compound mixtures and their concentrations.

[0118]

[0119]

[0120] 1 In all experiments, glucose was used as a positive control for growth and fermentation.

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131] 8 Duplicate E S. anginosus KC18-1131-3B had background growth of -0.04 log CFU / mL on MRS and -0.07 log CFU / mL on TSA, and L. crispatus KC18-1173-1 had background growth of 0.17 log CFU / mL on MRS.

[0132] 9 Replicate F S. anginosus KC18-1131-3B had a background growth of 0.32 log CFU / mL on MRS and 0.54 log CFU / mL on TSA, and L. crispatus KC18-1173-1 had a background growth of 0.17 log CFU / mL on MRS.

[0133] 10 Replicate G S. anginosus KC18-1131-3B had a background growth of -0.46 log CFU / mL on MRS and -0.39 log CFU / mL on TSA, and L. crispatus KC18-1173-1 had a background growth of 0.33 log CFU / mL on MRS.

[0134] 11 The background growth of Duplicate H Streptococcus anginosus KC18-1131-3B was -0.03 log CFU / mL on MRS and 0.09 log CFU / mL on TSA, and the background growth of Lactobacillus crispatus KC18-1173-1 was 0.35 log CFU / mL on MRS.

[0135] 12 Replicate I S. anginosus KC18-1131-3B had a background growth of 0.83 log CFU / mL on MRS and 1.12 log CFU / mL on TSA, and L. crispatus KC18-1173-1 had a background growth of -0.56 log CFU / mL on MRS.

[0136] 13Replicate J S. anginosus KC18-1131-3B had a background growth of 0.70 log CFU / mL on MRS and 0.91 log CFU / mL on TSA, and L. crispatus KC18-1173-1 had a background growth of 0.68 log CFU / mL on MRS.

[0137] 14 The background growth of replicate K S. anginosus KC18-1131-3B was 0.07 log CFU / mL on MRS and 0.07 log CFU / mL on TSA, and the background growth of Lactobacillus crispatus KC18-1173-1 was 0.40 log CFU / mL on MRS.

[0138] 15 The background growth of replicate L. anginosus KC18-1131-3B was 0.55 log CFU / mL on MRS and 0.70 log CFU / mL on TSA, and the background growth of L. crispatus KC18-1173-1 was -0.09 log CFU / mL on MRS.

[0139] 16 The background growth of replicate M Streptococcus anginosus KC18-1131-3B was -0.05 log CFU / mL on MRS and 0.23 log CFU / mL on TSA, and the background growth of Lactobacillus crispatus KC18-1173-1 was 0.98 log CFU / mL on MRS.

[0140] 17 The background growth of replicate N S. anginosus KC18-1131-3B was -0.32 log CFU / mL on MRS and -0.26 log CFU / mL on TSA, and the background growth of L. crispatus KC18-1173-1 was 0.56 log CFU / mL on MRS.

[0141] References

[0142] 1. Li, J., Peed, LA, Joyner, CF, Vongsa, RA, Koenig, DW, & Bartell, RD (2015). Composition for maintaining lactobacillus dominance. United States Patent Publication No. US2018 / 0256615A1.

[0143] 2.Zeng,Z.,Zhang W.,&Wu,K.Shenzhen(2019).Composition for vagina andantimicrobial agent combination for use in preparing composition forvagina.United States Patent Publication No.US2019 / 0231748A1.

[0144] 3.Zeng,Z.,&Zhou,R.(2016).Vaginal composition and use thereof.UnitedStates Patent Publication No.US2016 / 0375045A1.

[0145] 4.Vongsa,R.A.,Minerath,R.A.,Busch,M.A.,Tan,J.,&Koenig,D.W.(2016).Invitro evaluation of nutrients that selectively confer a competitive advantageto lactobacilli.Beneficial Microbes,7,299–304.

[0146] 5.Akgül T,Karakan T.The role of probiotics in women with recurrenturinary tract infections.Turk J Urol.2018Sep;44(5):377-383.doi:10.5152 / tud.2018.48742.Epub 2018Sep 1.PMID:30487041;PMCID:PMC6134985.

[0147] 6.Weysser Felipe de Souza,Francisco Lucas Chaves Almeida,RuannJanser Soares de Castro,Hélia Harumi Sato,Isomaltulose:From origin toapplication and its beneficial properties–A bibliometric approach,FoodResearch International,Volume 155,2022.

[0148] 7.Barea-Alvarez M,Benito MT,Olano A,Jimeno ML,Moreno FJ.Synthesis andcharacterization of isomaltulose-derived oligosaccharides produced bytransglucosylation reaction of Leuconostoc mesenteroides dextransucrase.JAgric Food Chem.2014Sep 17;62(37):9137-44.

[0149] 8.Kashimura J,Nagai Y,Goda T.Inhibitory action of palatinose and itshydrogenated derivatives on the hydrolysis of alpha-glucosylsaccharides inthe small intestine.J Agric Food Chem.2008 Jul 23;56(14):5892-5.

[0150] 9.Li T,Liu Z,Zhang X,Chen X,Wang S.Local Probiotic Lactobacilluscrispatus and Lactobacillus delbrueckii Exhibit Strong Antifungal EffectsAgainst Vulvovaginal Candidiasis in a Rat Model.Front Microbiol.2019 May 8;10:1033.

[0151] 10. Verhelst R, Verstraelen H, Claeys G, Verschraegen G, Van Simaey L, DeGanck C, De Backer E, Temmerman M, Vaneechoutte M. Comparison between Gram stain and culture for the characterization of vaginal microflora: definition of adistinct grade that resembles grade I microflora and revised categorization of grade I microflora. BMC Microbiol. 2005 Oct 14;5:61.

[0152] 11. Wu L, Wu S, Qiu J, Xu C, Li S, Xu H. Green synthesis of isomaltulose from cane molasses by Bacillus subtilis WB800-pHA01-palI in a biologicmembrane reactor. Food Chem. 2017 Aug 15; 229:761-768.

[0153] 12.Chee,WJY,Chew,SY&Than,LTLVaginal microbiota and thepotential of Lactobacillus derivatives in maintaining vaginal health.MicrobCell Fact 19,203(2020)

[0154] Implementation

[0155] Embodiment 1: A prebiotic preparation comprising the prebiotic agents isomaltulose and 1-kestose.

[0156] Embodiment 2: The prebiotic preparation of embodiment 1, wherein the prebiotic preparation further comprises at least one additional prebiotic agent.

[0157] Embodiment 3: The prebiotic formulation of Embodiment 3, wherein the additional prebiotic agent is lactulose.

[0158] Embodiment 4: The prebiotic formulation of any one of Embodiments 1 to 3, wherein the prebiotic formulation further comprises a carrier.

[0159] Embodiment 5: The prebiotic formulation of any one of Embodiments 1 to 4, wherein the prebiotic agent comprises 0.01% w / v to 20% w / v of the prebiotic formulation.

[0160] Embodiment 6: The prebiotic formulation of embodiment 5, wherein the prebiotic agent comprises 1.0% w / v to 3.0% w / v.

[0161] Embodiment 7: The prebiotic formulation of embodiment 6, wherein the prebiotic agent accounts for 2.0% w / v.

[0162] Embodiment 8: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 1.2% w / v and the 1-kestose is present at 0.8% w / v.

[0163] Embodiment 9: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 1.3% w / v and the 1-kestose is present at 0.7% w / v.

[0164] Embodiment 10: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 0.8% w / v and the 1-kestose is present at 1.2% w / v.

[0165] Embodiment 11: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 0.7% w / v and the 1-kestose is present at 1.3% w / v.

[0166] Embodiment 12: The prebiotic formulation of Embodiment 6, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 2.0% w / v and the 1-kestose is present at 0.2% w / v.

[0167] Embodiment 13: The prebiotic formulation of Embodiment 6, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 2.0% w / v and the 1-kestose is present at 0.1% w / v.

[0168] Embodiment 14: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 1.5% w / v and the 1-kestose is present at 0.5% w / v.

[0169] Embodiment 15: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 1.0% w / v and the 1-kestose is present at 1.0% w / v.

[0170] Embodiment 16: The prebiotic formulation of Embodiment 7, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 0.5% w / v and the 1-kestose is present at 1.5% w / v.

[0171] Embodiment 17: A prebiotic preparation comprising the prebiotic agents isomaltulose and lactulose.

[0172] Embodiment 18: The prebiotic formulation of Embodiment 11, wherein the prebiotic formulation further comprises at least one additional prebiotic agent.

[0173] Embodiment 19: The prebiotic formulation of Embodiment 12, wherein the additional prebiotic agent is 1-kestose.

[0174] Embodiment 20: The prebiotic formulation of any one of Embodiments 11 to 13, wherein the prebiotic formulation further comprises a carrier.

[0175] Embodiment 21: The prebiotic formulation of any one of Embodiments 11 to 14, wherein the prebiotic agent comprises 0.01% w / v to 20% w / v of the prebiotic formulation.

[0176] Embodiment 22: The prebiotic formulation of Embodiment 15, wherein the prebiotic agent comprises 1.0% w / v to 3.0% w / v.

[0177] Embodiment 23: The prebiotic formulation of Embodiment 16, wherein the prebiotic agent comprises 2.0% w / v.

[0178] Embodiment 24: The prebiotic formulation of Embodiment 17, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 1.5% w / v and the lactulose is present at 0.5% w / v.

[0179] Embodiment 25: The prebiotic formulation of Embodiment 23, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 0.7% w / v and the lactulose is present at 1.3% w / v.

[0180] Embodiment 26: The prebiotic formulation of Embodiment 23, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 1.0% w / v and the lactulose is present at 1.0% w / v.

[0181] Embodiment 27: The prebiotic formulation of Embodiment 23, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 0.5% w / v and the lactulose is present at 1.5% w / v.

[0182] Embodiment 28: The prebiotic formulation of Embodiment 22, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 2.0% w / v and the lactulose is present at 1.0% w / v.

[0183] Embodiment 29: The prebiotic formulation of Embodiment 22, wherein the isomaltulose is present in the prebiotic formulation at a concentration of 2.0% w / v and the lactulose is present at 0.1% w / v.

[0184] Embodiment 21: A method of modulating the growth of urogenital microorganisms in a subject, the method comprising administering a prebiotic formulation to the subject, wherein the prebiotic formulation comprises a prebiotic agent, wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestose and lactulose.

[0185] Embodiment 22: The method of Embodiment 21, wherein the prebiotic formulation further comprises a carrier.

[0186] Embodiment 23: The method of embodiment 22, wherein the carrier is a cream, liquid, solution, spray, paste or gel.

[0187] Embodiment 24: The method of Embodiment 23, wherein the carrier is water.

[0188] Embodiment 25: The method of any one of Embodiments 21 to 24, wherein the prebiotic formulation increases the growth of at least one beneficial bacteria.

[0189] Embodiment 26: The method of embodiment 25, wherein the beneficial bacteria are Lactobacillus.

[0190] Embodiment 27: The method of embodiment 26, wherein the beneficial bacteria is Lactobacillus crispatus.

[0191] Embodiment 28: The method of any one of Embodiments 21 to 25, wherein the prebiotic formulation reduces the growth of at least one pathogenic bacteria.

[0192] Embodiment 29: The method of Embodiment 28, wherein the pathogenic bacteria is Streptococcus anginosus.

[0193] Embodiment 30: The method of any one of Embodiments 21 to 29, wherein the ratio of isomaltulose to at least one of 1-kestose and lactulose is 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:5.

[0194] Embodiment 31: The method of any one of Embodiments 21 to 30, wherein the prebiotic formulation is administered topically.

[0195] Embodiment 32: The method of embodiment 31, wherein the prebiotic formulation has been applied in a matrix.

[0196] Embodiment 33: A method as described in Embodiment 32, wherein the matrix is ​​an absorbent material.

[0197] Embodiment 34: The method of Embodiment 33, wherein the absorbent material is a wipe, sheet, pad, or diaper.

[0198] Embodiment 35: A method of treating or preventing urogenital dysbacteriosis, the method comprising administering a prebiotic preparation to the subject, wherein the prebiotic preparation comprises a prebiotic agent, wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestose and lactulose.

[0199] Embodiment 36: The method of embodiment 35, wherein the subject suffers from, or is at risk of developing, overactive bladder, urge urinary incontinence, or a urinary tract infection.

[0200] Embodiment 37: The method of embodiment 36, wherein the subject has been diagnosed with overactive bladder, urge urinary incontinence, or a urinary tract infection.

[0201] Embodiment 38: The method of Embodiment 36, wherein the subject has been diagnosed as being at risk for developing overactive bladder, urge incontinence, or a urinary tract infection.

[0202] Embodiment 39: The method of any one of Embodiments 35 to 38, wherein the prebiotic formulation further comprises a carrier.

[0203] Embodiment 40: The method of Embodiment 39, wherein the carrier is a cream, liquid, solution, spray, paste or gel.

[0204] Embodiment 41: A method as described in Embodiment 40, wherein the carrier is water.

[0205] Embodiment 42: The method of any one of Embodiments 35 to 41, wherein the ratio of isomaltulose to at least one of 1-kestose and lactulose is.

[0206] Embodiment 43: The method of any one of Embodiments 35 to 42, wherein the prebiotic formulation is administered topically.

[0207] Embodiment 44: The method of Embodiment 43, wherein the prebiotic formulation has been applied in a matrix.

[0208] Embodiment 45: A method as described in Embodiment 44, wherein the matrix is ​​an absorbent material.

[0209] Embodiment 46: The method of Embodiment 45, wherein the absorbent material is a wipe, a sheet, a pad, or a diaper.

Claims

1. A prebiotic preparation comprising the prebiotic agents isomaltulose and 1-kestose.

2. The prebiotic preparation of claim 1, wherein the prebiotic preparation further comprises at least one additional prebiotic agent.

3. The prebiotic formulation of claim 2, wherein the additional prebiotic agent comprises lactulose.

4. The prebiotic preparation of claim 1, wherein the prebiotic preparation further comprises a carrier.

5. The prebiotic preparation of claim 1, wherein the prebiotic agent comprises 0.01% w / v to 20% w / v of the prebiotic preparation.

6. A prebiotic preparation comprising the prebiotic agents isomaltulose and lactulose.

7. The prebiotic formulation of claim 6, wherein the prebiotic formulation further comprises at least one additional prebiotic agent.

8. The prebiotic formulation of claim 7, wherein the additional prebiotic agent comprises 1-kestose.

9. The prebiotic preparation of claim 6, wherein the prebiotic preparation further comprises a carrier.

10. The prebiotic preparation of claim 6, wherein the prebiotic agent comprises 0.01% w / v to 20% w / v of the prebiotic preparation.

11. A method of modulating the growth of urogenital microorganisms in a subject, the method comprising administering to the subject a prebiotic formulation, wherein the prebiotic formulation comprises a prebiotic agent, wherein the prebiotic agent comprises isomaltulose and at least one of 1-kestose and lactulose.

12. The method of claim 11, wherein the prebiotic formulation further comprises a carrier, wherein the carrier is a cream, a liquid, a solution, a soap, a spray, a paste, or a gel.

13. The method of claim 11, wherein the prebiotic formulation increases the growth of at least one beneficial bacteria.

14. The method of claim 13, wherein the beneficial bacteria is Lactobacillus crispatus.

15. The method of any one of claims 11, wherein the prebiotic formulation reduces the growth of at least one pathogenic bacteria.

16. The method of claim 15, wherein the pathogenic bacteria is Streptococcus anginosus.

17. The method of claim 11, wherein isomaltulose and at least one of 1-kestose and / or lactulose are present in a ratio of 20:1, 13:7, 10:1, 5:1, 4:1, 3:1, 3:2, 2:1, 1:1, 1:2, 2:3, 1:3, 1:4, 1:5 or 7:

13.

18. The method of claim 11, wherein the prebiotic formulation is administered topically.

19. The method of claim 11, wherein the prebiotic formulation has been applied to a matrix, wherein the matrix is ​​an absorbent material.

20. The method of claim 19, wherein the absorbent material is a wipe, sheet, pad or diaper.

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