Patent Application
20220395481
The present invention relates to a bacteriostatic composition and a method for inhibiting harmful microorganisms. The present invention also relates to a method for modulating vaginal flora, a use of fatty acids and/or salts thereof in the preparation of the bacteriostatic composition for modulating vaginal flora, and a preparation method of the bacteriostatic composition.
Exposed to the external environment, human skin and mucosa are easily contaminated with pathogens, which may lead to pathogen colonization and infection of skin and mucosa, or lead to the carry and spread of pathogens. Cleaning skin and/or mucosa with bacteriostatic products can effectively reduce the pathogen load on skin and/or mucosa and even kill pathogens, thus helping to prevent the infection and spread of pathogens.
To prevent the spoilage of various personal cleaning and care products, such as skin care products, cosmetics, bath products, and shampoo products, as well as various medical products and pharmaceutical products, as a result of microbial contamination during their storage and use, it is essential to inhibit the growth of contaminated microorganisms and even to kill them, such as inhibiting or killing molds, yeast, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus.
In particular, the surface of the vaginal mucosa is inhabited by a large number of bacteria, fungi, and other microorganisms. As the most abundant bacteria in vagina of a healthy female, lactobacilli metabolize glycogens in vaginal mucosal epithelial cells, produce acids, maintain the vaginal pH in the range between 3.5-4.5, and produce inhibitory substances such as hydrogen peroxide and bacteriocin against pathogens and opportunistic pathogens. They are therefore beneficial and are known as “normal vaginal flora”.
The vagina is also inhabited by Gardnerella, Prevotella, Mobiluncus, Escherichia coli, Staphylococcus, Candida, and the like, which are not pathogenic when in small numbers, but may produce harmful metabolites, toxins, etc. when in large numbers thus resulting in pathological changes and diseases in human body. These microorganisms are therefore called “opportunistic pathogens”. Among Staphylococcus, the pathogenicity of Staphylococcus aureus is relatively high and representative. Among Candida infections, Candida albicans infection is the most common and representative, accounting for more than 70% of total vaginal and/or vulvar Candida infections.
When the vaginal flora is abnormal and lactobacilli exist in low numbers, the vagina is not only inhabited by a higher number of opportunistic pathogens such as Gardnerella, but also has a decreased resistance to pathogens of high toxicity and pathogenicity. The risk of communicable diseases of the reproductive tract, such as those caused by gonococci, Trichomonas, Chlamydia, mycoplasmas, and viruses such as HIV and HPV, and the like, then increases.
Currently, the antibacterial therapy remains the primary treatment for vaginal microbial diseases. Although antibacterial therapy can inhibit or kill pathogenic bacteria, it often inhibits or kills beneficial lactobacilli also and results in the reduction of vaginal resistance to infections. Thus, recurrent or persistent vaginal infections occur. It is a hot topic in medical research how to protect the beneficial bacteria in vagina during the course of antibacterial therapy and improve the efficacy of the prevention and treatment.
Therefore, there is a huge practical demand for a safe and effective bacteriostatic product, which is not only suitable for use in cleaning, hygiene, and bacteriostasis of human skin and mucosa, in the antisepsis of personal cleaning and care products such as bath products, skin care products, and cosmetics, and in the antisepsis of medical products, pharmaceutical products, and the like, but also can be used to inhibit abnormal vaginal flora, to restore and/or maintain vaginal lactobacilli, to restore and/or maintain normal vaginal flora, and to restore and/or maintain normal vaginal microecology.
The information in the Background section is intended only to describe the general context of the present invention and shall not be considered as an acknowledgment or in any way as an implication that such information constitutes the prior art known to those of ordinary skill in the art.
An objective of the present invention is to provide a bacteriostatic composition.
Another objective of the present invention is to provide a method for inhibiting harmful microorganisms.
A further objective of the present invention is to provide a method for modulating vaginal flora, referring to inhibiting abnormal vaginal flora, and/or restoring and/or maintaining vaginal lactobacilli.
A further objective of the present invention is to provide the use of fatty acids and/or salts thereof in the preparation of the bacteriostatic composition for modulating vaginal flora.
And a further objective of the present invention is to provide a method for preparing the bacteriostatic composition.
The present invention provides a bacteriostatic composition, wherein the composition includes the following ingredients:
(1) one or more of the fatty acids and/or salts thereof selected from the group consisting of acetic acid, glycolic acid, lactic acid, propionic acid, levulinic acid, butyric acid, isobutyric acid, hydroxybutyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof, preferably selected from the group consisting of propionic acid, butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, and salts thereof; the total content of the ingredient(s) (1) as described, calculated as fatty acid, is in the range of 0.001-3.00% (w/w);
(2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof, preferably adipic acid, and salt thereof; he total content of the ingredient(s) (2) as described, calculated as dicarboxylic acid, is in the range of 0.05-5.00% (w/w);
(3) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol, preferably selected from the group consisting of phenethyl alcohol and cinnamyl alcohol; the total content of the ingredient(s) (3) as described, is in the range of 0.03-1.00% (w/w);
(4) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, and salts thereof, preferably selected from the group consisting of benzoic acid, cinnamic acid, p-hydroxybenzoic acid, and salts thereof; the total content of the ingredient(s) (4) as described, calculated as aromatic carboxylic acid, is in the range of 0.03-1.00% (w/w);
The bacteriostatic composition is in one of the following dosage forms: aqueous solutions, water-soluble gels, foams, sprays, ointments, powders, films, capsules, suppositories, tablets, preferably aqueous solutions, water-soluble gels, foams, sprays, or ointments.
In some embodiments, the total content of one or more of the fatty acids and/or salts thereof, calculated as fatty acid, is preferably in the range of 0.001-2.00% (w/w), more preferably in the range of 0.001-1.50% (w/w), and the most preferably in the range of 0.002%-1.25% (w/w); and/or the total content of one or more of the dicarboxylic acids and/or salts thereof, calculated as dicarboxylic acid, is preferably in the range of 0.10-3.50% (w/w), more preferably in the range of 0.50-2.50% (w/w), and the most preferably in the range of 0.50-1.50% (w/w); and/or the total content of one or more of the aromatic alcohols is preferably in the range of 0.03-0.70% (w/w), more preferably in the range of 0.05-0.60% (w/w), and the most preferably in the range of 0.08-0.50% (w/w); and/or the total content of one or more of the aromatic carboxylic acids and/or salts thereof, calculated as aromatic carboxylic acids, is preferably in the range of 0.03-0.50% (w/w), more preferably in the range of 0.05-0.25% (w/w), and the most preferably in the range of 0.08-0.20% (w/w).
In some embodiments, the bacteriostatic composition further includes one or more of the dicarboxylic acids, polybasic carboxylic acids, and salts thereof selected from the group consisting of malic acid, citric acid, succinic acid, tartaric acid, maleic acid, isocitric acid, suberic acid, azelaic acid, sebacic acid, and salts thereof, preferably selected from the group consisting of malic acid, citric acid, succinic acid, and salts thereof; the total content of one or more of the dicarboxylic acids, polybasic carboxylic acids, and salts thereof as described is in the range of 0.10-2.50% (w/w). Wherein the acids and/or salts thereof have a buffering effect, which improves the pH stability of the bacteriostatic composition in the present disclosure, and further strengthens the modulating effect of the bacteriostatic composition on the pH of human skin and/or mucosa.
In some embodiments, the bacteriostatic composition can optionally include one or more of the monosaccharides and/or oligosaccharides and/or polysaccharides selected from the group consisting of glucose, fructose, mannose, galactose, maltose, isomaltose, sucrose, isomaltulose, lactose, lactulose, trehalose, cellobiose, melibiose, gentiobiose, 1-kestose, nystose, 1F-fructofuranosylnystose, isomaltotriose, isomaltotetraose, isomaltopentaose, gentiooligosaccharide, raffinose, panose, maltooligosaccharide, palatinose-oligosaccharide, oligofructose, glucomannan, galactooligosaccharide, dextrin, starch, and glycogen; the total content of which is in the range of 0.01-20.00% (w/w). The preferable saccharides are selected from the group consisting of isomaltulose, lactose, lactulose, maltose, isomaltose, trehalose, glycogen, and mixtures thereof; the total content of which is in the range of 0.10-2.00% (w/w). The saccharide-containing bacteriostatic composition in the present disclosure can not only inhibit Candida, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Gardnerella, Prevotella, Mobiluncus, Aspergillus niger, abnormal flora of skin and/or mucosa, and the like, but also can restore and/or maintain lactic acid bacteria on skin and/or mucosa, and restore and/or maintain the acidity of skin and/or mucosa. Therefore, it is suitable for use in the prevention and/or treatment of the decrease of lactic acid bacteria on skin and/or mucosa and the imbalance of skin and/or mucosa flora, such as the use in the prevention and/or treatment of bacterial vaginosis, aerobic vaginitis, or atrophic vaginitis, or the use in the adjuvant treatment of vaginitis, or the use in the recovery after vaginitis treatment, or the use in the treatment of reproductive tract infections, and the like.
In some embodiments, the bacteriostatic composition can optionally include one or more of the amino acids and/or salts thereof selected from the group consisting of L-glutamic acid, glutamine, L-aspartic acid, asparagine, leucine, isoleucine, phenylalanine, valine, proline, threonine, and salts thereof; the total content of which is in the range of 0.10-6.00% (w/w); the preferable amino acids are glutamic acid, aspartic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.50-3.00% (w/w). The bacteriostatic composition in the present disclosure containing amino acids and/or salts thereof can reduce acids production from lactic acid bacteria, thus decreasing the acidity of skin and/or mucosa. It is suitable for use in the modulation of the microenvironment of skin and/or mucosa, such as the use in the treatment or adjuvant treatment of cytolytic vaginosis, vulvovaginal candidiasis, and the like.
In some embodiments, the bacteriostatic composition can optionally include one or more of monoterpene and/or sesquiterpene compounds, including but not limited to citronellol, linalool, geraniol, nerol, eucalyptol, terpineol, carveol, menthol, and lavandulol; the total content of the monoterpene and/or sesquiterpene compounds as described is in the range of 0.001-0.50% (w/w). The monoterpene and/or sesquiterpene compounds may enhance the bacteriostatic effect of the bacteriostatic composition as well as provide a fragrance for the composition.
In some embodiments, the bacteriostatic composition can optionally include one or more of plant aromatic oils, including but not limited to rose essential oil, clove oil, red thyme oil, lavender oil, peppermint oil, Artemisia leaf oil, Eucalyptus oil, Sassafras oil, Litsea cubeba oil, cinnamon essential oil, laurel leaf oil, and thyme oil; the total content of the plant aromatic oils as described is in the range of 0.0001-0.05% (w/w). The plant aromatic oils can enhance the bacteriostatic effect of the composition, as well as provide a fragrance for the composition.
In some embodiments, the bacteriostatic composition can optionally include one or more of the vitamins selected from the group consisting of vitamin A, vitamin C, vitamin D, and vitamin E; the total content of which is in the range of 0.001-0.50% (w/w). The vitamins can be highly pure vitamins or the vitamins contained in plant extracts. The vitamins as described have effects of antioxidation, promoting the growth of skin cells, modulating the immune function, or maintaining the integrity of mucosal epithelial cells, etc., and can enhance the stability of the bacteriostatic composition, or protect human skin and/or mucosa.
In some embodiments, the bacteriostatic composition can optionally include one or more of the ingredients selected from the group consisting of dehydroacetic acid, sodium dehydroacetate, sorbic acid, potassium sorbate, sodium sorbate, natamycin, bergenin, tropolone, cinnamaldehyde, pseudolaric acid, chlorogenic acid, 1,2-pentanediol, 1,2-hexanediol, 1,6-hexanediol, 1,2-octanediol, 1,2-decanediol, p-hydroxyacetophenone, 2,4-dihydroxyacetophenone, methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, lysozyme, glycerol monocaprylate, glycerol monodecanoate, and glycerol monolaurate. The ingredients as described are used to further enhance the bacteriostatic or antibacterial effect of the bacteriostatic composition in the present disclosure against Candida, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Gardnerella, Prevotella, Mobiluncus, Aspergillus niger, abnormal flora of skin and/or mucosa, and the like, and other harmful microorganisms including but not limited to viruses such as HIV and HPV.
In some embodiments, the bacteriostatic composition further includes one or more of antibacterial drugs, including but not limited to nitroimidazoles such as metronidazole, tinidazole, and ornidazole; aminoglycosides such as gentamicin, tobramycin, amikacin, sisomicin, and netilmicin; quinolones such as ciprofloxacin, ofloxacin, and levofloxacin; furanes such as nifuratel, nifuroxime, furacilin, furazolidone, and furantoin; sulfonamides such as silver sulfadiazine and sodium sulfacetamide; pyrroles such as clotrimazole, fluconazole, miconazole, and ketoconazole; acrylamides such as naftifine and terbinafine; polyenes such as amphotericin B, nystatin, levorin, and natamycin; preferably metronidazole, nifuratel, clotrimazole, or mixtures thereof. The bacteriostatic composition in the present disclosure containing antibacterial drugs is suitable for skin and/or mucosa use, such as for the prevention and/or treatment of bacterial and/or fungal infections of skin, the prevention and/or treatment of bacterial or fungal infections of the oral mucosa, and the prevention and/or treatment of vaginal mucosa infections such as bacterial vaginosis, aerobic vaginitis, and vulvovaginal candidiasis.
In some embodiments, the bacteriostatic composition as described is in one of the following dosage forms: aqueous solutions, water-soluble gels, foams, sprays, or ointments; the pH value of the aqueous solutions, water-soluble gels, foams, sprays, or ointments is in the range of 3.1-4.8, preferably in the range of 3.6-4.6, and more preferably in the range of 3.8-4.4.
In some embodiments, the bacteriostatic composition is a water-soluble gel. The water-soluble gel includes one or more of non-flowable, viscous, and water-soluble colloidal excipients, including but not limited to xanthan gum, carbomer, polycarbophil, dextran, glucomannan, tragacanth gum, gummitragacanthae, methyl cellulose (MC), carboxymethyl cellulose (CMC), hydroxyethyl cellulose (HEC), hydroxyethyl methyl cellulose (HEMC), and hydroxypropyl methyl cellulose (HPMC), the preferred are xanthan gum and carbomer.
The bacteriostatic composition in the present disclosure has an inhibitory effect on harmful microorganisms, which is referred to but not limited to Candida, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Gardnerella, Prevotella, Mobiluncus, Aspergillus niger, abnormal flora of skin and/or mucosa, as well as various other pathogenic or opportunistically pathogenic aerobic bacteria, facultative bacteria, anaerobic bacteria, molds, and viruses such as HPV and HIV.
Therefore, the bacteriostatic composition in the present disclosure can be used for the cleaning and/or bacteriostasis and/or disinfection of human skin and/or mucosa such as vaginal mucosa, for the antisepsis of various personal cleaning and care products such as bath products, shampoo products, skin care products and cosmetics, or for the antisepsis of medical products, pharmaceutical products, and the like.
In some embodiments, the bacteriostatic composition in the present disclosure can be a therapeutic product, an active ingredient thereof, and an antiseptic thereof; the therapeutic product form includes but are not limited to drugs, disinfectants, topical microbicides, antibacterial agents, bacteriostatic agents, microecological modulators, flora modulators, microenvironment modulators, microbial modulators, disposable medical supplies, and the like, or the components of medical devices, the components of pharmaceutical devices, the components of disinfection devices, and the components of devices for vagina use.
In some embodiments, the bacteriostatic composition in the present disclosure can be a non-therapeutic product, an active ingredient thereof, and an antiseptic thereof; the non-therapeutic product form is one of the following group: health care products, hygiene products, personal cleaning and care products, cosmetics, disposable hygiene products, cleaning products, daily necessities, microecological care products, deodorants, lubricants, humectants, lotions, cleaning agents, body care products, antipruritic agents, refreshing agents, and the components of sanitary napkins, sanitary pads, and tampons.
In some embodiments, the bacteriostatic composition of the present invention is a vaginal bacteriostatic composition, wherein the vaginal bacteriostatic composition includes:
(1) one or more of the fatty acids and/or salts thereof selected from the group consisting of acetic acid, glycolic acid, lactic acid, propionic acid, levulinic acid, butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w). The preferred fatty acids and/or salts are selected from the group consisting of propionic acid, butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-1.50% (w/w), more preferably in the range of 0.002-1.25% (w/w);
(2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof; the total content of which, calculated as dicarboxylic acids, is in the range of 0.50-2.50% (w/w). The preferred is adipic acid and/or salt thereof; the total content of which, calculated as adipic acid, is in the range of 0.50-1.50% (w/w);
(3) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol; the total content of which is in the range of 0.05-0.60% (w/w). The preferred aromatic alcohols are phenethyl alcohol, cinnamic alcohol, and mixtures thereof; the total content of which is in the range of 0.08-0.50% (w/w);
(4) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, and salts thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.05-0.25% (w/w). The preferred aromatic carboxylic acids and/or salts thereof are selected from the group consisting of benzoic acid, cinnamic acid, p-hydroxybenzoic acid, salts thereof, and mixtures thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.08-0.20% (w/w);
The vaginal bacteriostatic composition as described is in one of the following dosage forms: aqueous solutions, water-soluble gels, foams, sprays, ointments, powders, films, capsules, suppositories, or tablets, preferably aqueous solutions, water-soluble gels, foams, sprays, or ointments.
In some embodiments, the vaginal bacteriostatic composition in the present disclosure further includes one or more of the estrogens selected from the group consisting of diethylstilbestrol, hexoestrol, estradiol, estrone, estriol, nilestriol, ethinyloestradiol, quinestrol, mestranol, and promestriene, preferably, estriol and promestriene; the total content of which is in the range of 0.001-1.00% (w/w). The vaginal bacteriostatic composition in the present disclosure containing estrogens can promote the glycogen synthesis in vaginal mucosal epithelial cells, and promote the growth of beneficial lactobacilli. It is especially suitable for use in menopause, postmenopausal, and postpartum.
In some embodiments, the vaginal bacteriostatic composition in the present disclosure further includes one or more of the phytoestrogens selected from the group consisting of daidzin, daidzein, glycitein, puerarin, coumestrol, genistein, equol, apigenin, genistin, genisteol, biochanin, coumestrol, formononetin, resveratrol, secoisolariciresinol, and lignan; the total content of which is in the range of 0.001-1.00% (w/w). The vaginal bacteriostatic composition in the present disclosure containing phytoestrogens can promote glycogen synthesis in vaginal mucosal epithelial cells, and promote the growth of beneficial lactobacilli. It is especially suitable for use in menopause, postmenopausal, and postpartum.
According to practical needs, the vaginal bacteriostatic composition in the present disclosure can optionally contain different excipients for different dosage forms such as water, xanthan gum, or carbomer. It can also optionally include one or more of the aforementioned ingredients further, such as the dicarboxylic acid(s), polybasic carboxylic acid(s), and salt(s) thereof, e.g., malic acid, citric acid, succinic acid, and salts thereof, in a total content of 0.10-2.50% (w/w); and/or the antibacterial or bacteriostatic agent(s) such as dehydroacetic acid and glycerol monocaprylate; and/or the saccharide(s) such as isomaltulose, lactose, and maltose, in a total content of 0.01-20.00% (w/w); and/or amino acid(s) such as glutamic acid, aspartic acid, and salts thereof, in a total content of 0.10-6.00% (w/w); and/or the monoterpene or sesquiterpene compound(s) such as citronellol and linalool, in a total content of 0.001-0.50% (w/w); and/or the plant aromatic oil(s) such as rose essential oil and clove oil, in a total content of 0.0001-0.05% (w/w); and/or the vitamin(s) such as vitamin A, and vitamin C, in a total content of 0.001-0.50% (w/w); and/or antibacterial drug(s) such as metronidazole, nifuratel, and clotrimazole.
Having an inhibitory effect on abnormal vaginal flora but an effect of restoring and/or maintaining and/or promoting vaginal lactobacilli, the vaginal bacteriostatic composition in the present disclosure can be used to restore and/or maintain normal vaginal flora, to restore and/or maintain normal vaginal microecology, and to restore and/or maintain normal vaginal acidity.
Therefore, the vaginal bacteriostatic composition in the present disclosure can be used to clean and take care of vagina and/or vulva, and/or to reduce and/or eliminate vaginal pruritus, and/or soreness, and/or dryness, and/or irritation, and/or dyspareunia, and/or to reduce and/or eliminate abnormal vaginal discharge, and/or unpleasant odor of vaginal discharge. It can also be used for the prevention and/or treatment and/or adjuvant treatment of the imbalance of vaginal flora, and/or bacterial vaginosis, and/or aerobic vaginitis, and/or cytolytic vaginosis, and/or vulvovaginal Candidiasis, and/or atrophic vaginitis, etc.
The present invention also provides a method for inhibiting harmful microorganisms, wherein the method includes the steps of using a bacteriostatic composition, wherein the bacteriostatic composition includes the following ingredients:
(1) one or more of the fatty acids and/or salts thereof selected from the group consisting of acetic acid, glycolic acid, lactic acid, propionic acid, levulinic acid, butyric acid, isobutyric acid, hydroxybutyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof, preferably selected from the group consisting of propionic acid, butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, and salts thereof; the total content of the ingredient(s) (1) as described, calculated as fatty acid, is in the range of 0.001-3.00% (w/w), preferably in the range of 0.001-2.00% (w/w), more preferably in the range of 0.001-1.50% (w/w), and the most preferably in the range of 0.002%-1.25% (w/w);
(2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof, preferably adipic acid, salt thereof; the total content of the ingredient(s) (2) as described, calculated as dicarboxylic acid, is in the range of 0.05-5.00% (w/w), preferably in the range of 0.10-3.50% (w/w), more preferably in the range of 0.50-2.50% (w/w), and the most preferably in the range of 0.50-1.50% (w/w);
(3) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol, preferably phenethyl alcohol, cinnamyl alcohol, and mixtures thereof; the total content of the ingredient(s) (3) as described is in the range of 0.03-1.00% (w/w), preferably in the range of 0.03-0.70% (w/w), more preferably in the range of 0.05-0.60% (w/w), and the most preferably in the range of 0.08-0.50% (w/w);
(4) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, and salts thereof, preferably selected from the group consisting of benzoic acid, cinnamic acid, p-hydroxybenzoic acid, and salts thereof; the total content of the ingredient(s) (4) as described, calculated as aromatic carboxylic acid, is in the range of 0.03-1.00% (w/w), preferably in the range of 0.03-0.50% (w/w), more preferably in the range of 0.05-0.25% (w/w), and the most preferably in the range of 0.08-0.20% (w/w);
Wherein the bacteriostatic composition is in one of the following dosage forms: aqueous solutions, water-soluble gels, foams, sprays, ointments, powders, films, capsules, suppositories, or tablets, preferably aqueous solutions, water-soluble gels, foams, sprays, or ointments.
In some embodiments, the harmful microorganisms as described in the method for inhibiting harmful microorganisms in the present disclosure refer to but are not limited to at least one of the following microorganisms: Candida, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Gardnerella, Prevotella, Mobiluncus, Aspergillus niger, abnormal flora of skin and/or mucosa, as well as other various pathogenic or opportunistically pathogenic aerobic bacteria, facultative bacteria, anaerobic bacteria, molds, and viruses such as HPV and HIV.
Therefore, the bacteriostatic composition as described in the method for inhibiting harmful microorganisms in the present disclosure can be used not only for cleaning and/or bacteriostasis and/or disinfection of human skin and/or mucosa, such as for cleaning and/or bacteriostasis and/or disinfection of vaginal mucosa, but also for the antisepsis of various personal cleaning and care products such as bath products, shampoo products, skin care products, and cosmetics, or for the antisepsis of medical products, pharmaceutical products, and the like.
In some embodiments, the bacteriostatic composition as described in the method for inhibiting harmful microorganisms in the present disclosure can be a therapeutic product, an active ingredient thereof, and an antiseptic thereof; wherein the therapeutic product form includes but are not limited to drugs, disinfectants, antibacterial agents, bacteriostatic agents, topical microbicides, flora modulators, microecological modulators, microenvironment modulators, microbial modulators, disposable medical supplies, the components of medical devices, the components of pharmaceutical devices, the components of disinfection devices, and the components of vaginal devices.
In some embodiments, the bacteriostatic composition as described in the method for inhibiting harmful microorganisms in the present disclosure can be a non-therapeutic product, an active ingredient thereof, or an antiseptic thereof; wherein the non-therapeutic product form includes but are not limited to health care products, personal cleaning and care products, cosmetics, hygiene products, disposable hygiene products, cleaning products, daily necessities, microecological care products, deodorants, lubricants, humectants, lotions, cleaning agents, body care products, antipruritic agents, refreshing agents, or can be the components of hygiene products or cleaning care products such as sanitary napkins, sanitary pads, or tampons.
When the bacteriostatic composition as described in the method in the present disclosure for inhibiting harmful microorganisms is used for antisepsis, the content of each ingredient of the bacteriostatic composition in the present disclosure contained in bath products, shampoo products, skin care products, cosmetics, pharmaceutical products, and other medical products, and the like, i.e. the total content of one or more of the fatty acids and/or salts thereof calculated as fatty acid, the total content of one or more of the dicarboxylic acids and/or salts thereof calculated as dicarboxylic acid, the total content of one or more of the aromatic alcohols, the total content of one or more of the aromatic carboxylic acids and/or salts thereof calculated as aromatic carboxylic acid, shall be within the total content ranges of the four ingredients in the bacteriostatic composition in the present disclosure.
The present invention also provides a method for modulating vaginal flora, wherein the method for modulating vaginal flora refers to at least one of the following: inhibiting abnormal vaginal flora, restoring and/or maintaining vaginal lactobacilli. Wherein the method includes the steps of using a vaginal bacteriostatic composition, wherein the vaginal bacteriostatic composition includes one or more of the fatty acids and/or salts thereof selected from the group consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w), preferably in the range of 0.001-1.50% (w/w). The more preferable fatty acids and/or salts thereof are selected from the group consisting of butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.002-1.25% (w/w);
Wherein the vaginal bacteriostatic composition is in one of the following dosage forms: aqueous solutions, water-soluble gels, foams, sprays, ointments, powders, films, capsules, suppositories, or tablets, preferably aqueous solutions, water-soluble gels, foams, sprays, or ointments.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present invention also includes one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof; the total content of which, calculated as dicarboxylic acid, is in the range of 0.50-2.50% (w/w). The preferable dicarboxylic acid and/or salt thereof is adipic acid and/or salt thereof, the total content of which is in the range of 0.50-1.50% (w/w). The combination of dicarboxylic acids such as adipic acid, salts thereof as described and fatty acids such as butyric acid, salts thereof in the present disclosure can enhance the bacteriostatic effect of the fatty acids and/or salts thereof as described against Candida albicans.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present invention also includes one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol; the total content of which is in the range of 0.05-0.60% (w/w). The preferable aromatic alcohols are phenethyl alcohol, cinnamyl alcohol, or mixtures thereof; the total content of which is in the range of 0.08-0.50% (w/w). The combination of aromatic alcohols such as phenethyl alcohol as described and fatty acids such as butyric acid, and salts thereof in the present disclosure can enhance the bacteriostatic effect of the fatty acids and/or salts thereof as described against Candida albicans, Staphylococcus aureus, Escherichia coli, abnormal vaginal flora, and the like.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present invention also includes one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, and salts thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.05-0.25% (w/w). The preferable aromatic carboxylic acids and/or salts thereof are selected from the group consisting of benzoic acid, cinnamic acid, p-hydroxybenzoic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.08-0.20% (w/w). The combination of aromatic carboxylic acids such as benzoic acid, salts thereof as described and fatty acids such as butyric acid, salts thereof in the present disclosure can enhance the bacteriostatic effect of the fatty acids and/or salts thereof as described against Candida albicans, Staphylococcus aureus, Escherichia coli, abnormal vaginal flora, and the like.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present invention includes: (1) one or more of the fatty acids and/or salts thereof selected from the group consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w), preferably in the range of 0.001-1.50% (w/w). The preferable fatty acids and salts thereof are selected from the group consisting of butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.002-1.25% (w/w); (2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof; the total content of which, calculated as dicarboxylic acid, is in the range of 0.50-2.50% (w/w); preferably adipic acid and/or salt thereof, the total content of which is in the range of 0.50-1.50% (w/w); (3) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol; the total content of which is in the range of 0.05-0.60% (w/w); the preferable aromatic alcohols are phenethyl alcohol, cinnamyl alcohol, or mixtures thereof; the total content of which is in the range of 0.08-0.50% (w/w).
The combination of dicarboxylic acids such as adipic acid and/or salts thereof, aromatic alcohols such as phenethyl alcohol, and fatty acids such as butyric acid and/or salts thereof in the present disclosure has a synergetic bacteriostatic effect against Candida albicans, Staphylococcus aureus, and Escherichia coli, and has an inhibitory effect on abnormal vaginal flora.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present disclosure includes: (1) one or more of the fatty acids and/or salts thereof selected from the group consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w), preferably in the range of 0.001-1.50% (w/w). The preferable fatty acids and/or salts thereof are selected from the group consisting of butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.002-1.25% (w/w); (2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof; the total content of which, calculated as dicarboxylic acid, is in the range of 0.50-2.50% (w/w); preferably adipic acid and/or salt thereof in the total content of 0.50-1.50% (w/w); (3) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, and salts thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.05-0.25% (w/w). The preferable aromatic carboxylic acids and/or salts thereof are benzoic acid, cinnamic acid, p-hydroxybenzoic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.08-0.20% (w/w).
The combination of dicarboxylic acids such as adipic acid and/or salts thereof, aromatic carboxylic acids such as benzoic acid and/or salts thereof, and fatty acids such as butyric acid and/or salts thereof in the present disclosure has a synergetic bacteriostatic effect against Candida albicans, Staphylococcus aureus, and Escherichia coli, and has an inhibitory effect on abnormal vaginal flora.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present disclosure includes: (1) one or more of the fatty acids and/or salts thereof selected from the group consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w), preferably in the range of 0.001-1.50% (w/w). The preferable fatty acids and/or salts thereof are selected from the group consisting of butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.002-1.25% (w/w); (2) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol; the total content of which is in the range of 0.05-0.60% (w/w). The preferable aromatic alcohols are phenethyl alcohol, cinnamyl alcohol, or mixtures thereof; the total content of which is in the range of 0.08-0.50% (w/w); (3) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, salts thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.05-0.25% (w/w). The preferable aromatic carboxylic acids and/or salts thereof are selected from the group consisting of benzoic acid, cinnamic acid, p-hydroxybenzoic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.08-0.20% (w/w).
The combination of aromatic alcohols such as phenethyl alcohol, aromatic carboxylic acids such as benzoic acid and/or salts thereof, and fatty acids such as butyric acid and/or salts thereof in the present disclosure has a synergetic bacteriostatic effect against Candida albicans, Staphylococcus aureus, and Escherichia coli, and has an inhibitory effect on abnormal vaginal flora.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present disclosure includes: (1) one or more of the fatty acids and/or salts thereof selected from the group consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w), preferably in the range of 0.001-1.50% (w/w). The preferable fatty acids and/or salts thereof are selected from the group consisting of butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.002-1.25% (w/w); (2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof; the total content of which, calculated as dicarboxylic acid, is in the range of 0.50-2.50% (w/w); preferably adipic acid and/or salt thereof in the total content of 0.50-1.50% (w/w); (3) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol; the total content of which is in the range of 0.05-0.60% (w/w). The preferable aromatic alcohols are phenethyl alcohol, cinnamyl alcohol, or mixtures thereof; the total content of which is in the range of 0.08-0.50% (w/w); (4) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, salts thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.05-0.25% (w/w). The preferable aromatic carboxylic acids and/or salts thereof are selected from the group consisting of benzoic acid, cinnamic acid, p-hydroxybenzoic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.08-0.20% (w/w).
The combination of dicarboxylic acids such as adipic acid and/or salts thereof, aromatic alcohols such as phenethyl alcohol, aromatic carboxylic acids such as benzoic acid and/or salts thereof, and fatty acids such as butyric acid and/or salts thereof in the present disclosure has a synergetic bacteriostatic effect against Candida albicans, Staphylococcus aureus, and Escherichia coli, and has an inhibitory effect on abnormal vaginal flora. After vaginal application of the bacteriostatic composition, there was a significant decrease in abnormal vaginal flora and a significant increase in lactobacilli. Therefore, the bacteriostatic composition as described has the effects of inhibiting abnormal vaginal flora, restoring and/or maintaining and/or promoting vaginal lactobacilli, and restoring and/or maintaining normal vaginal acidity.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present disclosure further includes one or more of the estrogens selected from the group consisting of diethylstilbestrol, hexoestrol, estradiol, estrone, estriol, nilestriol, ethinyloestradiol, quinestrol, mestranol, and promestriene; the total content of which is in the range of 0.001-1.00% (w/w), preferably estriol and promestriene. As estrogens can promote the glycogen synthesis of vaginal mucosal epithelial cells and promote the growth of beneficial lactobacilli, the bacteriostatic composition in the present disclosure containing estrogens is especially suitable for use in menopause, postmenopausal, or postpartum.
In some embodiments, the vaginal bacteriostatic composition as described in the method for modulating vaginal flora in the present disclosure further includes one or more of the phytoestrogens selected from the group consisting of daidzin, daidzein, glycitein, puerarin, coumestrol, genistein, equol, apigenin, genistin, genisteol, biochanin, coumestrol, formononetin, resveratrol, secoisolariciresinol, and lignan; the total content of which is in the range of 0.001-1.00% (w/w). As phytoestrogens can promote the glycogen synthesis of vaginal mucosal epithelial cells and promote the growth of beneficial lactobacilli, the bacteriostatic composition in the present disclosure containing phytoestrogens is especially suitable for use in menopause, postmenopausal, or postpartum.
In some embodiments, the method for modulating vaginal flora in the present invention includes the use of the vaginal bacteriostatic composition to restore and/or maintain normal vaginal flora, and/or to restore and/or maintain normal vaginal microecology, and/or to restore and/or maintain normal vaginal acidity, and/or to clean and take care of the vagina and/or vulva, and/or to reduce and/or eliminate vaginal pruritus, soreness, dryness, irritation, and dyspareunia, and/or to reduce and/or eliminate abnormal vaginal discharge and unpleasant odor of vaginal discharge.
In some embodiments, the method for modulating vaginal flora in the present invention includes the use of the vaginal bacteriostatic composition to prevent and/or treat the imbalance of vaginal flora, bacterial vaginosis, aerobic vaginitis, cytolytic vaginosis, vulvovaginal Candidiasis, and/or atrophic vaginitis.
The present invention also provides the use of fatty acids and/or salts thereof in the preparation of the bacteriostatic composition for modulating vaginal flora. The fatty acids and/or salts thereof are at least one selected from the groups consisting of butyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-2.00% (w/w), preferably in the range of 0.001-1.50% (w/w). The preferable fatty acids and/or salts thereof are selected from the group consisting of butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof; the total content of which is in the range of 0.002-1.25% (w/w).
In some embodiments, the use of fatty acids and/or salts thereof in the preparation of the bacteriostatic composition for modulating vaginal flora according to the present invention, wherein the fatty acids and/or salts thereof are in the same concentration and properties as those of the fatty acids and/or salts thereof as described in the method for modulating vaginal flora in the present disclosure.
In some embodiments, the use of fatty acids and/or salts thereof in the preparation of a bacterial bacteriostatic composition for modulating vaginal flora according to the present invention, wherein the modulating vaginal flora refers to at least one of the following: inhibiting abnormal vaginal flora, and restoring and/or maintaining vaginal lactobacilli, the same as that of the method for modulating vaginal flora in the present disclosure.
In some embodiments, the use of fatty acids and/or salts thereof in the preparation of a bacterial bacteriostatic composition for modulating vaginal flora according to the present invention, wherein the modulating vaginal flora includes the use of the vaginal bacteriostatic composition to restore and/or maintain normal vaginal flora, and/or to restore and/or maintain normal vaginal microecology, and/or to restore and/or maintain normal vaginal acidity, and/or to clean and take care of the vagina and/or vulva, and/or to reduce and/or eliminate vaginal pruritus, soreness, dryness, irritation, and dyspareunia, and/or to reduce and/or eliminate abnormal vaginal discharge and unpleasant odor of vaginal discharge.
In some embodiments, the use of fatty acids and/or salts thereof in the preparation of a bacterial bacteriostatic composition for modulating vaginal flora according to the present invention, wherein modulating vaginal flora includes the use of the vaginal bacteriostatic composition to prevent and/or treat the imbalance of vaginal flora, bacterial vaginosis, aerobic vaginitis, cytolytic vaginosis, vulvovaginal Candidiasis, and atrophic vaginitis.
The present invention also provides a preparation method of the bacteriostatic composition, wherein the preparation method includes the following steps: to add the following ingredients to an excipient, wherein the excipient is used for aqueous solutions, water-soluble gels, foams, suppositories, or tablets:
(1) one or more of the fatty acids and/or salts thereof selected from the group consisting of acetic acid, glycolic acid, lactic acid, propionic acid, levulinic acid, butyric acid, isobutyric acid, hydroxybutyric acid, valeric acid, hexanoic acid, heptanoic acid, caprylic acid, nonanoic acid, capric acid, undecylic acid, undecylenic acid, lauric acid, and salts thereof; the total content of which, calculated as fatty acid, is in the range of 0.001-3.00% (w/w); preferably propionic acid, butyric acid, hexanoic acid, caprylic acid, capric acid, undecylic acid, salts thereof, and mixtures thereof;
(2) one or more of the dicarboxylic acids and/or salts thereof selected from the group consisting of glutaric acid, adipic acid, pimelic acid, and salts thereof; the total content of which, calculated as dicarboxylic acid, is in the range of 0.05-5.00% (w/w); preferably adipic acid, and salt thereof;
(3) one or more of the aromatic alcohols selected from the group consisting of benzyl alcohol, 2,4-dichlorobenzene methanol, phenethyl alcohol, phenoxyethanol, and cinnamyl alcohol; the total content of which is in the range of 0.03-1.00% (w/w); preferably phenethyl alcohol, cinnamic alcohol, or mixtures thereof;
(4) one or more of the aromatic carboxylic acids and/or salts thereof selected from the group consisting of benzoic acid, p-hydroxybenzoic acid, p-methoxybenzoic acid, salicylic acid, cinnamic acid, gentianic acid, caffeic acid, and salts thereof; the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.03-1.00% (w/w); preferably benzoic acid, cinnamic acid, p-hydroxybenzoic acid, salts thereof, and mixtures thereof.
The composition as described is in one of the following dosage forms: aqueous solutions, water-soluble gels, foams, sprays, ointments, powders, films, capsules, suppositories, or tablets, preferably, aqueous solutions, or water-soluble gels, or foams, or sprays, or ointments.
In some embodiments, the preparation method includes the addition of one or more of the fatty acids and/or salts thereof as described, the total content of which, calculated as fatty acid, is preferably in the range of 0.001-2.00% (w/w), more preferably in the range of 0.001-1.50% (w/w), and the most preferably in the range of 0.002%-1.25% (w/w); and/or the addition of one or more of the dicarboxylic acids and/or salts thereof as described, the total content of which, calculated as dicarboxylic acid, is preferably in the range of 0.10-3.50% (w/w), more preferably in the range of 0.50-2.50% (w/w), and the most preferably in the range of 0.50-1.50% (w/w); and/or the addition of one or more of the aromatic alcohols, the total content of which is preferably in the range of 0.03-0.70% (w/w), more preferably in the range of 0.05-0.60% (w/w), and the most preferably in the range of 0.08-0.50% (w/w); and/or the addition of one or more of the aromatic carboxylic acids, the total content of which is preferably in the range of 0.03-0.50% (w/w), more preferably in the range of 0.05-0.25% (w/w), and the most preferably in the range of 0.08-0.20% (w/w).
In the preparation of water-soluble gels, a non-flowable, viscous, and water-soluble colloidal excipient is needed, which enables the composition to homogeneously contact with the vaginal mucosa and to stay for a longer time to take effect. In some embodiments, wherein the excipient is xanthan gum, carbomer, polycarbophil, dextran, glucomannan, tragacanth gum, gummitragacanthae, methyl cellulose (MC), carboxymethyl cellulose (CMC), hydroxyethyl cellulose (HEC), hydroxyethyl methyl cellulose (HEMC), and hydroxypropyl methyl cellulose (HPMC), preferably xanthan gum, and carbomer.
In some embodiments, wherein the composition can be prepared according to the following technological process: weigh and take (1) one or more of the fatty acids and/or salts thereof such as propionic acid and/or sodium salt thereof, (2) one or more of the dicarboxylic acids and/or salts thereof such as adipic acid and/or salts thereof, (3) one or more of the aromatic alcohols such as phenethyl alcohol, (4) one or more of the aromatic carboxylic acids and/or salts thereof such as benzoic acid and/or sodium salts thereof, (5) one or more of the colloidal substrates such as xanthan gum, and other ingredients, mix well, add purified water quantitatively, stir and mix well, so that each ingredient is dissolved and the colloidal substrate swells into homogeneous colloid; adjust the pH value of the composition with acid and/or alkali to the range of 3.1-4.8, preferably 3.6-4.6, and more preferably 3.8-4.4. In some embodiments, sterilization can be further carried out, wherein the sterilization technology may be selected from the following: radiation sterilization, high-temperature sterilization (for example, sterilizing at 115.6° C. for 15-20 minutes; or sterilizing at 100° C. for >30 minutes), intermittent sterilization (for example, sterilizing at 80° C. for 30 minutes, then placing at 36° C. for 5-10 hours, then once again at 80° C. for 30 minutes, then at 36° C. for 5-10 hours, and finally at 80° C. for 30 minutes). Alternatively, in some embodiments, wherein ingredients such as benzoic acid and/or sodium salt thereof are dissolved separately into solutions, sterilized by filtration, and then added into a sterilized water-soluble gel.
In some embodiments, when a solution is prepared, all of the above ingredients except xanthan gum can be mixed well, dissolved with water, sterilized, and packaged; or dissolved, filtered, and packaged.
In some embodiments, when an emulsion-type ointment is prepared, the selection of excipient and the specific preparation process can be referred to methods known to those skilled in the art, such as the method introduced in Pharmacy (see Reference 1) edited by Fang Liang.
In some embodiments, a tablet can be prepared by referring to methods known to those skilled in the art, such as the method introduced in Pharmacy (see Reference 2) edited by Fang Liang, where quantified fatty acid and/or salt thereof, dicarboxylic acid and/or salt thereof, aromatic alcohol, aromatic carboxylic acid and/or salt thereof, and other ingredients are thoroughly mixed with excipients, then directly tableting. Optionally, excipients can be added, e.g., lubricants such as magnesium stearate or disintegrants such as sodium carboxymethyl starch can be added, homogeneously mixed, then tableting. In some embodiments, the prepared tablets can also be packaged into administration apparatus, disinfection devices, medical devices, or pharmaceutical devices.
In some embodiments, a film or suppository can be prepared by referring to methods known to those skilled in the art, such as the method introduced in Pharmacy (see References 3 and 4) edited by Fang Liang.
In some embodiments, when sanitary napkins, sanitary pads, or tampons containing the bacteriostatic composition in the present disclosure are prepared, ingredients such as fatty acid and/or salt thereof, dicarboxylic acid and/or salt thereof, aromatic alcohol, aromatic carboxylic acid and/or salt thereof in the present disclosure, e.g., sodium benzoate, sodium propionate, hexanoic acid, phenethyl alcohol, adipic acid, and other ingredients, can be weighed and taken, then added with corresponding excipients, and prepared into powders, films, tablets, or capsules, which are then placed inside the sanitary napkins, sanitary pads, or tampons through an appropriate method or process. Optionally, ingredients such as fatty acid and/or salt thereof, dicarboxylic acid and/or salt thereof, aromatic alcohol, aromatic carboxylic acid and/or salt thereof in the present disclosure, such as sodium propionate, hexanoic acid, phenethyl alcohol, sodium benzoate, adipic acid, and other ingredients, and appropriate excipients, are attached to the inner-layer materials of sanitary napkins, sanitary pads, or tampons via an appropriate process.
In some embodiments, when the dosage forms such as capsules, suppositories, or tablets are prepared, each unit dosage form includes a specific content range of the ingredients of the composition in the present disclosure. For example, each unit dosage form includes one or more of the fatty acids and/or salts thereof, the total content of which, calculated as fatty acid, is in the range of 0.00005-0.15 g; one or more of the dicarboxylic acids and/or salts thereof, the total content of which, calculated as dicarboxylic acid and/or polybasic carboxylic acid, is in the range of 0.0025-0.25 g; one or more of the aromatic alcohols, the total content of which is in the range of 0.0015-0.050 g; and one or more of the aromatic carboxylic acids and/or salts thereof, the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.0015-0.05 g. Preferably, each unit dosage form includes one or more of the fatty acids and/or salts thereof, the total content of which, calculated as fatty acid, is in the range of 0.00005-0.100 g; one or more of the dicarboxylic acids and/or salts thereof, the total content of which, calculated as dicarboxylic acid and/or polybasic carboxylic acid, is in the range of 0.005-0.175 g; one or more of the aromatic alcohols, the total content of which is in the range of 0.0015-0.035 g; and one or more of the aromatic carboxylic acids and/or salts thereof, the total content of which, calculated as aromatic carboxylic acid, is in the range of 0.0015-0.025 g.
Various organic acids and/or salts thereof used in the preparation method in the present disclosure, such as fatty acid and/or salt thereof, dicarboxylic acid and/or salt thereof, or aromatic carboxylic acid and/or salt thereof, are dissolved in water into non-ionized molecules and ionized ions. The ratio of the non-ionized molecules to the ionized ions depends on the pH value of the solutions and the ionization constant (pKa) of the organic acid. Evidently, both organic acids and salts thereof are organic acid molecules or organic acid radical ions after dissolution in water. From this perspective, there is no substantive difference between the two.
To make the aforementioned more comprehensible, several embodiments accompanied with drawings are described in detail as follows.
Multiple exemplary embodiments of the present invention are described in detail below. This detailed description should not be considered a limitation to the invention but a more detailed description of certain aspects, features, and embodiments of the invention. It should be understood that the terms in the present invention are intended to describe particular embodiments only, rather than to limit the invention. Furthermore, wherein a range of values is provided in this invention, it should be understood that the upper and lower limits of that range, and each intermediate value between them, are also specifically disclosed. Each smaller range between any stated value or intermediate value in a stated range and any other stated value or intermediate value in that stated range is included in the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All literature cited in the specification is incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the literature. In case of conflict with any incorporated literature, this Specification shall prevail. Unless otherwise stated, “%” is a percentage by weight.
Bacteriostatic agents, commonly used for the cleaning, bacteriostasis, or disinfection of skin and/or mucosa such as povidone iodine and chlorhexidine, are mostly broad-spectrum bacteriostatic agents that inhibit not only Escherichia coli, Staphylococcus aureus, and fungi, but also lactic acid bacteria such as lactobacilli. Antiseptics, commonly used in skin care products, cosmetics, bath products, as well as medical products and pharmaceutical products such as chlorobutanol and benzalkonium chloride, usually also have an inhibitory effect on lactic acid bacteria such as lactobacilli. There is a lack of highly selective bacteriostatic agents or bacteriostatic compositions that strongly inhibit harmful bacteria, but weakly inhibit beneficial bacteria such as lactobacilli.
The prior art (e.g., PCT/CN2017/105296, ZL201080036139, and U.S. Pat. No. 8,765,819) discloses compositions formulated with the combination of low-concentration bacteriostatic agents, such as the low-concentration of phenethyl alcohol, propionic acid and/or salt thereof, and/or benzoic acid and/or salt thereof, which can inhibit Escherichia coli and Staphylococcus aureus, with an inhibition rate against Escherichia coli and Staphylococcus aureus of up to more than 50% and a low inhibition rate against Candida albicans of <50% according to the test methods and judgment criteria of results in the National Standards of the People's Republic of China Hygienic Standard for Disposable Sanitary Products (GB15979-2002).
Increasing the concentration of the bacteriostatic agent can enhance the bacteriostatic effect against Candida albicans. Studies have shown that an appropriate increase in the concentration of each ingredient in the bacteriostatic combination of “propionic acid and/or salt thereof+benzoic acid and/or salt thereof+phenethyl alcohol” can not only enhance the bacteriostatic effect of the combination of bacteriostatic agents, therefor more effectively inhibit Escherichia coli, Staphylococcus aureus, and the like, but also inhibit Candida albicans by an inhibition rate of more than 50%. However, the higher the concentration of the bacteriostatic agent is, the stronger the inhibition against lactobacilli and other lactic acid bacteria becomes. The experimental example I disclosed in PCT/CN2017/105296 showed that when the concentration of sodium propionate was 0.40% (w/v) or 0.50% (w/v), the growth of lactobacilli was not significantly inhibited; when the concentration of sodium propionate was 0.70% (w/v), the growth of lactobacilli was inhibited. When the concentration of phenethyl alcohol was 0.40% (w/v), the growth of lactobacilli was not significantly inhibited; when the concentration of phenethyl alcohol was 0.50% (w/v), the growth of lactobacilli and acid production was inhibited.
The inventor has continuously carried out an in-depth study to further develop bacteriostatic compositions that effectively inhibit harmful microorganisms but not beneficial bacteria such as lactobacilli, and that can be used for the cleaning, bacteriostasis, or disinfection of skin and/or mucosa, and can be used as antiseptics for skin care products, cosmetics, and bath products, as well as medical products and pharmaceutical products. The inventor has found that the selective combination of appropriate fatty acids and/or salts thereof with dicarboxylic acids, aromatic alcohols, and aromatic carboxylic acids has a synergetic bacteriostatic effect against not only Staphylococcus aureus and Escherichia coli, but also Candida albicans, as well as significantly inhibits the growth of Pseudomonas aeruginosa and Aspergillus niger. The vaginal use of bacteriostatic composition prepared accordingly can result in a significant decrease in abnormal vaginal flora and a significant increase in lactobacilli, thus effectively modulate vaginal flora and vaginal acidity. Therefore, the bacteriostatic composition in the present disclosure can be used for the cleaning, bacteriostasis, or disinfection of skin and/or mucosa, and for antisepsis of bath products, skin care products, and cosmetics, as well as medical products, and pharmaceutical products.
In order to facilitate the accurate understanding of relevant nouns or phrases or expressions or standards mentioned herein, the meanings thereof are hereby clarified and defined as follows:
“Lactobacillus” refers to the bacteria of the genus Lactobacillus, a kind of gram-positive, rod-shaped, non-spore-forming bacteria capable of producing a large amount of lactic acid as a by-product of glucose metabolism, including hundreds of species and subspecies.
“Lactic acid bacteria” is a general term of bacteria capable of metabolizing fermentable carbohydrates to produce a large amount of lactic acid, and refer to bacteria of more than 200 species of 18 genera. Lactobacillus is also a kind of lactic acid bacteria.
“Normal vaginal flora” refers to a Nugent score of 1-3. The vaginal bacteria are dominated by Lactobacillus species and with a small number of other bacteria. Wherein the “other bacteria” refer to gram-positive cocci such as Staphylococcus and Streptococcus, gram-negative bacilli such as Gardnerella and Escherichia coli, gram-negative cocci such as Veillonella parvula, and obligate anaerobes such as Prevotella and Mobiluncus.
“Abnormal vaginal flora” refers to a Nugent score of 5-10. The vaginal bacteria are dominated by other bacteria and with a small number of Lactobacillus species. Wherein the “other bacteria” refer to gram-positive cocci such as Staphylococcus and Streptococcus, gram-negative bacilli such as Gardnerella and Escherichia coli, gram-negative cocci such as Veillonella parvula, and obligate anaerobes such as Prevotella and Mobiluncus.
“Harmful microorganisms” generally refer to various highly pathogenic microorganisms, or various opportunistically pathogenic microorganisms that cause disease when human immunity is weakened, or microorganisms that cause the spoilage of food, pharmaceutical products, cosmetics, and hygiene products, including but not limited to the following group of microorganisms: Candida, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Gardnerella, Prevotella, Mobiluncus, Aspergillus niger, the abnormal flora of skin and/or mucosa, as well as viruses such as HPV and HIV.
“Ingredient” refers to various ingredients in the composition, including bacteriostatic agents and other ingredients without bacteriostatic activity.
“Bacteriostatic agent” refers to various ingredients with bacteriostatic effects, or combinations thereof, not limited to conventional bacteriostatic agents or combinations thereof recognized by those skilled in the art.
“Inhibition rate” is the inhibition rate of bacteriostatic agents against Escherichia coli, Staphylococcus aureus, Candida albicans, or other microorganisms obtained by referring to the test methods and judgment criteria in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products.
“Bacteriostatic effect” refers to the inhibition of bacteria, fungi, or other microorganisms by bacteriostatic agents. The strength of inhibition is judged herein based on the inhibition rate, specifically as follows: inhibition rate >50%, indicating “possessing bacteriostatic effect”; inhibition rate >90%, indicating “strong bacteriostatic effect”.
“Low-concentration bacteriostatic agent” is a relatively low concentration bacteriostatic agent with an inhibition rate of <50% against Candida albicans.
“High-concentration bacteriostatic agent” is a relatively high concentration bacteriostatic agent with an inhibition rate of >50% against Candida albicans.
“No difference in inhibition rates” or “no influence on bacteriostatic effect” indicates that the difference between two inhibition rates is <15%.
“A difference in inhibition rates” or “influence on bacteriostatic effect” indicates that the difference between two inhibition rates is 15-30%.
“A significant difference in inhibition rates” or “significant influence on bacteriostatic effect” indicates that the difference between two inhibition rates is >30%.
“Synergistic bacteriostatic effect” refers to the combination of two or more bacteriostatic agents acting against Escherichia coli, Staphylococcus aureus, or Candida albicans, which has an inhibition rate that is more than 15% higher than the sum of the inhibition rates of each bacteriostatic agent acting alone.
This embodiment provides a plurality of exemplary ingredients of the bacteriostatic composition. Unless specifically stated otherwise, exemplary ingredients of the composition, described hereunder, are the following substances:
Propionic acid CAS: 79-09-4, adipic acid CAS: 124-04-9, phenethyl alcohol (2-phenylethanol) CAS: 60-12-8, butyric acid (n-butyric acid) CAS: 107-92-6, hexanoic acid (n-hexanoic acid) CAS: 142-62-1, pimelic acid CAS: 111-16-0, malic acid (L-hydroxysuccinic acid) CAS: 97-67-6, cinnamic acid (trans-cinnamic acid) CAS: 140-10-3, salicylic acid (2-hydroxybenzoic acid) CAS: 69-72-7, lauric acid (dodecanoic acid) CAS: 143-07-7, valeric acid (n-valeric acid) CAS: 109-52-4, heptanoic acid (enanthic acid) CAS: 111-14-8, caprylic acid (n-caprylic acid) CAS: 124-07-2, nonanoic acid (n-nonanoic acid) CAS: 112-05-0, capric acid (n-capric acid) CAS: 334-48-5, undecylic acid (undecanoic acid) CAS: 112-37-8, cinnamyl alcohol (3-phenyl-2-propen-1-ol) CAS: 104-54-1, succinic acid (butanedioic acid) CAS: 110-15-6, tartaric acid (L-tartaric acid) CAS: 87-69-4, maleic acid (cis-butenedioic acid) CAS: 110-16-7, citric acid CAS: 77-92-9, fumaric acid (trans-butenedioic acid) CAS: 110-17-8, undecylenic acid (10-undecylenic acid) CAS: 112-38-9, glutaric acid CAS: 110-94-1.
Add 1.50 g of adipic acid, 2.00 g of propionic acid, 0.25 g of phenethyl alcohol, 1.00 g of citric acid, and 2.50 g of maltose to 80 g of purified water and stir to dissolve them, then add 2.15 g of xanthan gum, and complement purified water to render a total weight of 100 g. Stir for the xanthan gum to swell into a homogeneous viscous gel, then adjust pH to 3.1 with 1.0 mol/L of sodium hydroxide solution, and sterilize at 115.6° C. for 15 minutes, to obtain the water-soluble gel composition in the present disclosure.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of the composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of the composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of composition by basically following the method in example 1.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of the composition by basically following the method in example 1.
Adipic acid 1.50 g, benzoic acid 0.20 g, propionic acid 1.25 g;
Glutamic acid 0.76 g, glutamine 0.34 g, aspartic acid 0.60 g, asparagine 0.98 g, isoleucine 0.31 g, methionine 0.35 g, phenylalanine 0.20 g, valine 0.46 g, leucine 0.78 g, proline 0.89 g;
Xanthan gum 2.50 g;
add purified water to reach the total weight of 100 g, and adjust pH to 4.0.
Raw materials were weighed and taken according to the following formula, to prepare 100 g of the composition by basically following the method in example 1.
A tablet containing 60 mg of adipic acid, 5 mg of sodium benzoate, 30 mg of propionic acid, and 60 mg of sucrose was prepared basically following the method described in Reference 1.
A vaginal suppository containing 60 mg of adipic acid, 5 mg of sodium benzoate, 30 mg of propionic acid, and 60 mg of maltose was prepared basically following the method described in Reference 3.
This embodiment is used to validate the efficacy of the composition.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231. The percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 1.
Results:
1. It was evident in the result of Group 1 in Table 1 that, when the pH value was 3.8, the solution containing “0.50% propionic acid, 1.50% adipic acid, and 0.20% sodium benzoate” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 61.29%;
2. It was evident in the results of Groups 2-3 in Table 1 that, when the pH value was 3.8, the combinations of “0.50% propionic acid, 1.50% adipic acid, and 0.20% sodium benzoate” and benzyl alcohol of two different concentrations (0.50%, 0.70%) respectively, had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 81.52% and 99.27%, respectively.
In summary, it could be seen from the comparison of experimental results of Groups 2-3 and Group 1 that, when the pH value was 3.8, 0.50% and 0.70% benzyl alcohol respectively affected the bacteriostatic effect of the solution containing “0.50% propionic acid, 1.50% adipic acid, and 0.20% sodium benzoate” against Candida albicans, and could enhance the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231. The percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.0. The experimental results are shown in Table 2.
Results:
1. It was evident in the result of Group 1 in Table 2 that, when the pH value was 4.0, the inhibition rate of the solution containing “1.50% propionic acid, 0.35% phenethyl alcohol, and 0.20% benzoic acid” against Candida albicans was 15.91%;
2. It was evident in the result of Group 2 in Table 2 that, when the pH value was 4.0, the inhibition rate of the solution containing “1.50% propionic acid, 0.35% phenethyl alcohol, and 0.25% benzoic acid” against Candida albicans was 46.09%;
3. It was evident in the result of Group 3 in Table 2 that, when the pH value was 4.0, the solution containing “1.50% propionic acid, 1.50% adipic acid, 0.35% phenethyl alcohol, and 0.20% benzoic acid” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 67.39%;
4. It was evident in the result of Group 4 in Table 2 that, when the pH value was 4.0, the solution containing “1.50% propionic acid, 1.50% adipic acid, 0.35% phenethyl alcohol, and 0.25% benzoic acid” had a strong bacteriostatic effect against Candida albicans, and the inhibition rate was 92.17%.
In summary,
1. It was evident in the comparison of experimental results of Groups 1 and 2, and the result of Groups 3 and 4 that, when the pH value was 4.0, the increase in the concentration of benzoic acid from 0.20% to 0.25% affected or significantly affected the bacteriostatic effects of the solution containing “1.50% propionic acid and 0.35% phenethyl alcohol” and the solution containing “1.50% propionic acid, 1.50% adipic acid, and 0.35% phenethyl alcohol” against Candida albicans, and enhanced the bacteriostatic effect;
2. It was evident in the comparison of experimental results of Groups 1 and 3, and Groups 2 and 4 that, when the pH value was 4.0, 1.50% adipic acid significantly affected the bacteriostatic effect of the solution containing “1.50% propionic acid, 0.35% phenethyl alcohol, and 0.20% benzoic acid” and the solution containing “1.50% propionic acid, 0.35% phenethyl alcohol, and 0.25% benzoic acid” against Candida albicans, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 3.
Results:
1. It was evident in the result of Group 1 in Table 3 that, when the pH value was 3.8, the inhibition rate of the solution containing “1.35% adipic acid, 0.25% benzyl alcohol, and 0.20% sodium benzoate” against Candida albicans was 45.12%;
2. It was evident in the results of Groups 2-5 in Table 3 that, when the pH value was 3.8, the combinations of propionic acid of four different concentrations respectively (0.50%, 1.25%, 1.50%, 3.00%) and “1.35% adipic acid, 0.25% benzyl alcohol, and 0.20% sodium benzoate” had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 61.60%, 89.72%, 94.42%, and 100.00%, respectively.
In summary, it could be seen from the comparison of experimental results of Groups 2-5 and Group 1 that, when the pH value was 3.8, propionic acid affected or significantly affected the bacteriostatic effect of the solution containing “1.35% adipic acid, 0.25% benzyl alcohol, and 0.20% sodium benzoate” against Candida albicans, and enhanced the bacteriostatic effect. The higher the concentration of propionic acid was, the stronger the bacteriostatic effect against Candida albicans became.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.3. The experimental results are shown in Table 4.
Results:
1. It was evident in the result of Group 1 in Table 4 that when the pH value was 4.3, the inhibition rate of the solution containing “0.54% propionic acid, 1.50% adipic acid, and 0.30% phenethyl alcohol” against Candida albicans was 26.05%;
2. It was evident in the results of Groups 2 and 3 in Table 4 that when the pH value was 4.3, the combinations of “0.54% propionic acid, 1.50% adipic acid, and 0.30% phenethyl alcohol” with 0.50% or 1.00% benzoic acid respectively had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 64.80% and 100.00% respectively.
In summary, when the pH value was 4.3, 0.50% and 1.00% benzoic acid significantly affected the bacteriostatic effect of the solution containing “0.54% propionic acid, 1.50% adipic acid, and 0.30% phenethyl alcohol” against Candida albicans, and enhanced the bacteriostatic effect. The higher the concentration of benzoic acid was, the stronger the bacteriostatic effect against Candida albicans became.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.6. The experimental results are shown in Table 5.
Results:
1. It was evident in the result of Group 1 in Table 5 that when the pH value was 4.6, the solution containing “1.50% adipic acid, 0.30% phenethyl alcohol, and 1.18% sodium benzoate” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 84.12%;
2. It was evident from the result of Group 2 in Table 5 that, when the pH value was 4.6, the solution containing “2.00% propionic acid, 1.50% adipic acid, 0.30% phenethyl alcohol, and 1.18% sodium benzoate” had a strong bacteriostatic effect against Candida albicans, and the inhibition rate was 100.00%.
In summary, it could be seen from the comparison of experimental results of Group 1 and Group 2 that, when the pH value was 4.6, 2.00% propionic acid affected the bacteriostatic effect of the solution containing “1.50% adipic acid, 0.30% phenethyl alcohol, and 1.18% sodium benzoate” against Candida albicans, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 6.
Results:
1. It was evident in the results of Groups 1-2 in Table 6 that when the pH value was 3.8, the combinations of “0.20% sodium benzoate and 0.40% phenethyl alcohol” and two fatty acids of different concentrations (0.88% butyric acid, 0.12% hexanoic acid) respectively had a bacteriostatic effect against Candida albicans, and the inhibition rates were 71.52% and 70.65% respectively;
2. It was evident in the results of Groups 3-4 in Table 6 that when the pH value was 3.8, the solution containing “0.20% sodium benzoate, 0.40% phenoxyethanol, and 0.88% butyric acid” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 56.75%; the solution containing “0.20% sodium benzoate, 0.40% phenoxyethanol, and 0.12% hexanoic acid” had an inhibition rate of 35.88% against Candida albicans;
3. It was evident in the results of Groups 5-6 in Table 6 that, when the pH value was 3.8, the combinations of “1.46% adipic acid, 0.20% sodium benzoate, and 0.40% phenethyl alcohol” and two fatty acids of different concentrations (0.88% butyric acid, 0.12% hexanoic acid) respectively had a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 100%;
4. It was evident in the results of Groups 7-8 in Table 6 that when the pH value was 3.8, the combinations of “1.46% adipic acid, 0.20% sodium benzoate, and 0.40% phenoxyethanol” and two fatty acids of different concentrations (0.88% butyric acid, 0.12% hexanoic acid) respectively had a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 100%.
In summary,
1. It was evident in the comparison of experimental results of Groups 5-6 and Groups 1-2 in Table 6 that, when the pH value was 3.8, 1.46% adipic acid affected the bacteriostatic effect of the solution containing “0.20% sodium benzoate, 0.40% phenethyl alcohol, and 0.88% butyric acid” and the solution containing “0.20% sodium benzoate, 0.40% phenethyl alcohol, and 0.12% hexanoic acid” against Candida albicans respectively, and enhanced the bacteriostatic effect;
2. It was evident in the comparison of experimental results of Groups 7-8 and Groups 3-4 in Table 6 that, when the pH value was 3.8, 1.46% adipic acid significantly affected the bacteriostatic effect of the solution containing “0.20% sodium benzoate, 0.40% phenoxyethanol, and 0.88% butyric acid” and the solution containing “0.20% sodium benzoate, 0.40% phenoxyethanol, and 0.12% hexanoic acid” against Candida albicans respectively, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 7.
Candida albicans ATCC 10231
indicates data missing or illegible when filed
Results:
1. It was evident in the result of Group 1 in Table 7 that, when the pH value was 3.8, the solution containing “0.21% p-hydroxybenzoic acid, 0.06% phenethyl alcohol, and 2.50% propionic acid” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 62.76%;
2. It was evident in the result of Group 2 in Table 7 that, when the pH value was 3.8, the combination of 1.60% pimelic acid and “0.21% p-hydroxybenzoic acid, 0.06% phenethyl alcohol, and 2.50% propionic acid” had a strong bacteriostatic effect against Candida albicans, and the inhibition rate was 94.84%;
3. It was evident in the result of Group 3 in Table 7 that, when the pH value was 3.8, the combination of 1.34% malic acid and “0.21% p-hydroxybenzoic acid, 0.06% phenethyl alcohol, and 2.50% propionic acid” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 65.68%;
In summary, it could be seen from the comparison of experimental results of Groups 1 and 2, and Groups 1 and 3 that, when the pH value was 3.8, 1.60% pimelic acid significantly affected the bacteriostatic effect of the solution containing “0.21% p-hydroxybenzoic acid, 0.06% phenethyl alcohol, and 2.50% propionic acid” against Candida albicans, and enhanced the bacteriostatic effect, while 1.34% malic acid did not affect the bacteriostatic effect of the solution containing “0.21% p-hydroxybenzoic acid, 0.06% phenethyl alcohol, and 2.50% propionic acid” against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.9. The experimental results are shown in Table 8.
Results:
1. It was evident in the results of Groups 1-4 in Table 8 that, when the pH value was 3.9, four solutions containing 1.46% adipic acid, 0.60% butyric acid, 0.14% sodium benzoate, and 0.40% phenethyl alcohol respectively had inhibition rates of 11.52%, 1.57%, 19.90%, and −1.05% respectively against Candida albicans;
2. It was evident in the result of Group 5 in Table 8 that, when the pH value was 3.9, the solution containing “1.46% adipic acid and 0.60% butyric acid” had an inhibition rate of 13.61% against Candida albicans;
3. It was evident in the result of Group 6 in Table 8 that, when the pH value was 3.9, the solution containing “0.60% butyric acid and 0.14% sodium benzoate” had an inhibition rate of 17.80% against Candida albicans;
4. It was evident in the result of Group 7 in Table 8 that, when the pH value was 3.9, the solution containing “0.60% butyric acid and 0.40% phenethyl alcohol” had an inhibition rate of 8.90% against Candida albicans;
5. It was evident in the result of Group 8 in Table 8 that, when the pH value was 3.9, the solution containing “1.46% adipic acid, 0.60% butyric acid, and 0.14% sodium benzoate” had an inhibition rate of 27.23% against Candida albicans;
6. It was evident in the result of Group 9 in Table 8 that, when the pH value was 3.9, the solution containing “1.46% adipic acid, 0.60% butyric acid, and 0.40% phenethyl alcohol” had an inhibition rate of 25.65% against Candida albicans;
7. It was evident in the result of Group 10 in Table 8 that, when the pH value was 3.9, the solution containing “0.60% butyric acid, 0.40% phenethyl alcohol, and 0.14% sodium benzoate” had an inhibition rate of 2.62% against Candida albicans;
8. It was evident in the result of Group 11 in Table 8 that, when the pH value was 3.9, the solution containing “1.46% adipic acid, 0.14% sodium benzoate, and 0.40% phenethyl alcohol” had an inhibition rate of 42.41% against Candida albicans;
9. It was evident in the result of Group 12 in Table 8 that, when the pH value was 3.9, the solution containing “1.46% adipic acid, 0.60% butyric acid, 0.14% sodium benzoate, and 0.40% phenethyl alcohol” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 89.53%;
In summary, it could be seen that when the pH value was 3.9, the inhibition rate of the combination of 1.46% adipic acid, 0.60% butyric acid, 0.14% sodium benzoate, and 0.40% phenethyl alcohol against Candida albicans was much higher than that of each of the ingredients or the combination of any two or three of these ingredients, indicating the combination of these four ingredients had a synergistic bacteriostatic effect against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.3. The experimental results are shown in Table 9.
Results:
1. It was evident in the results of Groups 1˜4 in Table 9 that, when the pH value was 4.3, the inhibition rates against Candida albicans of four solutions containing 1.35% adipic acid, 0.20% sodium benzoate, 0.18% hexanoic acid, and 0.30% phenethyl alcohol respectively were all <10%;
2. It was evident in the result of Group 5 in Table 9 that, when the pH value was 4.3, the solution containing “1.35% adipic acid, 0.20% sodium benzoate, 0.18% hexanoic acid, and 0.30% phenethyl alcohol” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 58.86%;
3. It was evident from the results of Groups 6-9 in Table 9 that, when the pH value was 4.3, the combination of any three ingredients selecting from 1.35% adipic acid, 0.20% sodium benzoate, 0.18% hexanoic acid, and 0.30% phenethyl alcohol, had an inhibition rate of less than 50% against Candida albicans.
In summary, it could be seen that the inhibition rate of the combination of 1.35% adipic acid, 0.20% sodium benzoate, 0.18% hexanoic acid, and 0.30% phenethyl alcohol, against Candida albicans was significantly higher than that of the combination of any three of these four ingredients, and also significantly higher than the sum of the inhibition rate of each ingredient, indicating that the combination of these four ingredients had a synergistic bacteriostatic effect against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 10.
Candida albicans ATCC 10231
indicates data missing or illegible when filed
Results:
1. It was evident in the result of Group 1 in Table 10 that, when the pH value was 3.8, the solution containing 1.50% adipic acid, 0.45% phenethyl alcohol, and 1.50% propionic acid had an inhibition rate of 30.96% against Candida albicans;
2. It was evident in the results of Groups 2-5 in Table 10 that, when the pH value was 3.8, the solution combining cinnamic acids of four different concentrations (0.03%, 0.05%, 0.08%, 0.10%) respectively with the solution containing “1.50% adipic acid, 0.45% phenethyl alcohol, and 1.50% propionic acid” had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 59.04%, 71.19%, 96.38%, and 100.00% respectively;
3. It was evident in the results of Groups 6-9 in Table 10 that, when the pH value was 3.8, the combination of 0.03% salicylic acid with “1.50% adipic acid, 0.45% phenethyl alcohol, and 1.50% propionic acid” had an inhibition rate of 37.40% against Candida albicans, while the combination of salicylic acid of three higher concentrations (0.05%, 0.08%, 0.10%) respectively with “1.50% adipic acid, 0.45% phenethyl alcohol, and 1.50% propionic acid” had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 55.58%, 86.96%, and 94.81%, respectively.
In summary, it could be seen that 0.03-0.10% cinnamic acid and 0.05-0.10% salicylic acid affected or significantly affected the bacteriostatic effect of the solution containing “1.50% adipic acid, 0.45% phenethyl alcohol, and 1.50% propionic acid”, against Candida albicans. The higher the concentration of cinnamic acid or salicylic acid was, the further enhanced the bacteriostatic effect became.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 11.
Candida albicans ATCC 10231
Results:
1. It was evident in the results of Groups 1-6 in Table 11 that, when the pH value was 3.8, each one of 0.001-0.002% undecylic acid, 0.001-0.002% undecylenic acid, or 0.001-0.002% lauric acid respectively had inhibition rates of <15% against Candida albicans;
2. It was evident in the result of Group 7 in Table 11 that, when the pH value was 3.8, the solution containing “0.20% sodium benzoate, 0.25% phenethyl alcohol, and 1.50% adipic acid” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 50.02%;
3. It was evident in the results of Groups 8-9 in Table 11 that, when the pH value was 3.8, the combinations of “0.20% sodium benzoate, 0.25% phenethyl alcohol, and 1.50% adipic acid” and undecylic acid of two different concentrations (0.001%, 0.002%) respectively had a strong bacteriostatic effect against Candida albicans, and the inhibition rates were higher than 90%;
4. It was evident in the results of Groups 10-11 in Table 11 that, when the pH value was 3.8, the combinations of “0.20% sodium benzoate, 0.25% phenethyl alcohol, and 1.50% adipic acid” and undecylenic acid of two different concentrations (0.001%, 0.002%) respectively had a bacteriostatic effect against Candida albicans, and the inhibition rates were higher than 50%;
5. It was evident in the results of Groups 12-13 in Table 11 that, when the pH value was 3.8, the combinations of “0.20% sodium benzoate, 0.25% phenethyl alcohol, and 1.50% adipic acid” and lauric acid of two different concentrations (0.001%, 0.002%) respectively had a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 100%.
In summary, it could be seen that 0.001-0.002% undecylic acid, 0.002% undecylenic acid, and 0.001-0.002% lauric acid significantly affected the bacteriostatic effect of the solution containing “0.20% sodium benzoate, 0.25% phenethyl alcohol, and 1.50% adipic acid” against Candida albicans respectively, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, and the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w). The experimental results are shown in Tables 12-14.
albicans ATCC 10231
Results:
1. It was evident in the results in Table 12 that, when the pH value was 3.6-3.8, each one of the ten fatty acids of different concentrations (2.00% propionic acid, 2.20% butyric acid, 0.60% valeric acid, 0.20% hexanoic acid, 0.07% heptanoic acid, 0.02% caprylic acid, 0.009% nonanoic acid, 0.002% capric acid, 0.001% undecylic acid, 0.001% lauric acid) or 0.25% sodium benzoate respectively had inhibition rates of <20% against Candida albicans;
2. It was evident in the results in Table 13 that, when the pH value the 3.6-3.8, the combinations of 0.20-0.25% sodium benzoate with each one of the ten fatty acids of different concentrations (2.00% propionic acid, 2.20% butyric acid, 0.60% valeric acid, 0.20% hexanoic acid, 0.07% heptanoic acid, 0.02% caprylic acid, 0.009% nonanoic acid, 0.002% capric acid, 0.001% undecylic acid, 0.001% lauric acid) respectively had a bacteriostatic effect against Candida albicans, and the inhibition rates were >50%, indicating that the combination of these two ingredients had a synergistic bacteriostatic effect against Candida albicans, and enhanced the bacteriostatic effect;
3. It was evident in the results in Table 14 that, when the pH value was 3.6-3.8, the combinations of “0.20% sodium benzoate and 0.40% phenethyl alcohol” with each one of the ten fatty acids of different concentrations (2.00% propionic acid, 2.20% butyric acid, 0.60% valeric acid, 0.20% hexanoic acid, 0.07% heptanoic acid, 0.02% caprylic acid, 0.009% nonanoic acid, 0.002% capric acid, 0.001% undecylic acid, 0.001% lauric acid) respectively had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 50%-90% or >90%, indicating that the combination of these three ingredients had a synergistic bacteriostatic effect against Candida albicans, and enhanced the bacteriostatic effect;
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 15.
Results:
1. It was evident in the results of Groups 1-10 in Table 15 that, when the pH value was 3.8, each one of the nine fatty acids of different concentrations (2.00% propionic acid, 2.20% butyric acid, 0.60% valeric acid, 0.20% hexanoic acid, 0.07% heptanoic acid, 0.02% caprylic acid, 0.005% capric acid, 0.003% lauric acid, 0.003% undecylic acid) or 0.30% cinnamyl alcohol respectively had inhibition rates of <25% against Candida albicans;
2. It was evident in the results of Groups 11-19 in Table 15 that, when the pH value was 3.8, the combinations of each one of the nine fatty acids of different concentrations (2.00% propionic acid, 2.2% butyric acid, 0.60% valeric acid, 0.20% hexanoic acid, 0.07% heptanoic acid, 0.02% caprylic acid, 0.005% capric acid, 0.003% lauric acid, 0.003% undecylic acid) respectively with 0.30% cinnamyl alcohol had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 50%-90% or >90%, indicating that the combination of these two ingredients had a synergistic bacteriostatic effect against Candida albicans, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 16.
Results:
1. It was evident in the results of Groups 1-3 in Table 16 that, when the pH value was 3.8, each one of 1.00% butyric acid, 1.00% pimelic acid, or 0.60% phenethyl alcohol respectively had inhibition rates of <10% against Candida albicans;
2. It was evident in the result of Group 4 in Table 16 that, when the pH value was 3.8, the inhibition rate of the solution containing 1.00% butyric acid and 0.60% phenethyl alcohol against Candida albicans was 11.09%;
3. It was evident in the result of Group 5 in Table 16 that, when the pH value was 3.8, the solution containing 1.00% butyric acid, 1.00% pimelic acid, and 0.60% phenethyl alcohol had a bacteriostatic effect against Candida albicans, and the inhibition rate was 65.22%.
In summary, it could be seen that the solution containing 1.00% butyric acid, 1.00% pimelic acid, and 0.60% phenethyl alcohol had a synergistic bacteriostatic effect against Candida albicans, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing sodium propionate and dicarboxylic acid or tricarboxylic acid were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.6. The experimental results are shown in Table 17.
Results:
1. It was evident in the results of Groups 1-8 in Table 17 that, when the pH value was 3.6, each one of the 2.88% sodium propionate and seven dicarboxylic acids or polybasic carboxylic acids of different concentrations (4.38% adipic acid, 3.54% succinic acid, 4.02% malic acid, 4.50% tartaric acid, 3.50% maleic acid, 5.76% citric acid, 3.50% fumaric acid) respectively had inhibition rates of <25% against Candida albicans;
2. It was evident in the result of Group 9 in Table 17 that, when the pH value was 3.6, the solution containing 1.25% sodium propionate and 4.38% adipic acid had a bacteriostatic effect against Candida albicans, and the inhibition rate was 72.46%;
3. It was evident in the results of Groups 10-15 in Table 17 that, when the pH value was 3.6, the combinations of 1.25% sodium propionate and each one of the six dicarboxylic acids or polybasic carboxylic acids of different concentrations (3.54% succinic acid, 4.02% malic acid, 4.50% tartaric acid, 3.50% maleic acid, 5.76% citric acid, 3.50% fumaric acid) respectively had inhibition rates of <25% against Candida albicans.
In summary, it could be seen that the combination of adipic acid and sodium propionate had a synergistic bacteriostatic effect against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, the combination of citric acid, malic acid, or succinic acid respectively with a basic bacteriostatic solution containing “1.46% (w/w) adipic acid+0.16% (w/w) sodium benzoate+0.62% (w/w) propionic acid+0.35% (w/w) phenethyl alcohol” was studied for the bacteriostatic effect of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of the basic bacteriostatic solution was 3.6. The experimental results are shown in Table 18.
Results:
1. The basic bacteriostatic solution had a bacteriostatic effect against Candida albicans, and the inhibition rate was 53.77%;
2. The combinations of basic bacteriostatic solution with 1.00% citric acid, 0.70% malic acid, or 0.61% succinic acid respectively had inhibition rates of 49.25%, 54.77%, and 53.27% against Candida albicans respectively.
In summary, it could be seen that citric acid, malic acid, or succinic acid did not affect the bacteriostatic effect of the basic bacteriostatic solution containing “1.46% (w/w) adipic acid+0.16% (w/w) sodium benzoate+0.62% (w/w) propionic acid+0.35% (w/w) phenethyl alcohol” against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 19.
Results:
1. It was evident in the results of Groups 1-9 in Table 19 that, when the pH value was 3.8, each one of the seven fatty acids of different concentrations (1.32% butyric acid, 0.70% valeric acid, 0.20% hexanoic acid, 0.12% heptanoic acid, 0.03% caprylic acid, 0.0075% nonanoic acid, 0.005% capric acid) and two dicarboxylic acids of different concentrations (1.50% adipic acid, 2.00% pimelic acid) respectively had inhibition rates of <35% against Candida albicans;
2. It was evident in the results of Groups 10-16 in Table 19 that, when the pH value was 3.8, the combinations of each one of the seven fatty acids of different concentrations (1.32% butyric acid, 0.70% valeric acid, 0.20% hexanoic acid, 0.12% heptanoic acid, 0.03% caprylic acid, 0.0075% nonanoic acid, 0.005% capric acid) respectively with 1.50% adipic acid had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 50%-90% or >90%;
3. It was evident in the results of Groups 17-23 in Table 19 that, when the pH value was 3.8, the combinations of each one of the seven fatty acids of different concentrations (1.32% butyric acid, 0.70% valeric acid, 0.20% hexanoic acid, 0.12% heptanoic acid, 0.03% caprylic acid, 0.0075% nonanoic acid, 0.005% capric acid) respectively with 2.00% pimelic acid had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 50%-90% or >90%.
In summary, it could be seen that, when the pH value was 3.8, the combinations of each one of the seven fatty acids of different concentrations (1.32% butyric acid, 0.70% valeric acid, 0.20% hexanoic acid, 0.12% heptanoic acid, 0.03% caprylic acid, 0.0075% nonanoic acid, 0.005% capric acid) respectively and 1.50% adipic acid or 2.00% pimelic acid respectively had a synergistic bacteriostatic effect against Candida albicans, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538, and Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 20.
Candida
Escherichia
Staphylococcus
albicans
coli
aureus
It was evident in the results in Table 20 that, when the pH value was 3.8, each one of the nine fatty acids of different concentrations (3.00% butyric acid, 0.45% hexanoic acid, 0.30% heptanoic acid, 0.05% caprylic acid, 0.03% nonanoic acid, 0.008% capric acid, 0.006% undecylic acid, 0.006% undecylenic acid, 0.006% lauric acid) respectively and 5.00% pimelic acid had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 50%-90% or >90%, and had a strong bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were 100.00%.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 21.
Results:
1. It was evident in the result of Group 1 in Table 21 that, when the pH value was 3.8, the solution containing 1.50% pimelic acid, 0.20% sodium benzoate, and 2.00% propionic acid had an inhibition rate of 41.25% against Candida albicans;
2. It was evident in the results of Groups 2-4 in Table 21 that, when the pH value was 3.8, the combinations of cinnamyl alcohol of 0.03%, 0.05%, or 0.08% respectively with “1.50% pimelic acid, 0.20% sodium benzoate, and 2.00% propionic acid” had a bacteriostatic effect or a strong bacteriostatic effect against Candida albicans, and the inhibition rates were 50%-90% or >90% respectively.
In summary, it could be seen that each one of the cinnamyl alcohol of 0.03, 0.05%, or 0.08% significantly affected the bacteriostatic effect of the solution containing “1.50% pimelic acid, 0.20% sodium benzoate, and 2.00% propionic acid” against Candida albicans respectively, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.8. The experimental results are shown in Table 22.
Results:
1. It was evident in the results of Groups 1-3 in Table 22 that, when the pH value was 4.8, each one of 5.00% pimelic acid, 1.18% sodium benzoate, or 0.10% propionic acid respectively had inhibition rates of less than 20% against Candida albicans;
2. It was evident in the result of Group 4 in Table 22 that, when the pH value was 4.8, the solution containing “5.00% pimelic acid, 1.18% sodium benzoate, and 0.10% propionic acid” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 61.02%.
In summary, it could be seen that, when the pH value was 4.8, the solution containing 5.00% pimelic acid, 1.18% sodium benzoate, and 0.10% propionic acid had a synergistic bacteriostatic effect against Candida albicans, and enhanced the bacteriostatic effect.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their inhibition rates of 20 minutes of action against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 6538, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.0. The experimental results are shown in Table 23.
Escherichia
Staphylococcus
coli
aureus
Results:
1. It was evident in the results of Groups 1˜4 in Table 23 that, when the pH value was 4.0, the combinations of 0.06% sodium benzoate and each one of 0.30% phenethyl alcohol, 0.50% propionic acid, 0.45% butyric acid, or 0.06% hexanoic acid respectively had a bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were higher than 50%;
2. It was evident in the results of Groups 5-7 in Table 23 that, when the pH value was 4.0, the combinations of 0.30% phenethyl alcohol and each one of 0.50% propionic acid, 0.45% butyric acid, or 0.06% hexanoic acid respectively had a bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were higher than 50%;
3. It was evident in the results of Groups 8-10 in Table 23 that, when the pH value was 4.0, the combination of “0.045% sodium benzoate, 0.30% phenethyl alcohol” and 0.35% propionic acid had a bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were higher than 50%; the combinations of “0.045% sodium benzoate, 0.30% phenethyl alcohol” and 0.30% butyric acid or 0.06% hexanoic acid respectively had a strong bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were higher than 90%;
4. It was evident in the results of Groups 11-13 in Table 23 that, when the pH value was 4.0, the combinations of “1.50% adipic acid, 0.045% sodium benzoate, and 0.30% phenethyl alcohol” and each one of the three fatty acids of different concentrations (0.35% propionic acid, 0.30% butyric acid, 0.06% hexanoic acid) respectively had a strong bacteriostatic effect against Escherichia coli, and the inhibition rates were higher than 90%; and had a bacteriostatic effect against Staphylococcus aureus, and the inhibition rates were 50%-90%.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, and the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w). The experimental results are shown in Table 24.
Results:
It was evident in the results of Groups 1-3 in Table 24 that, when the pH value was 4.0 and 4.2 respectively, the solution containing “0.06% citric acid, 1.17% adipic acid, 0.20% propionic acid, 0.60% butyric acid, 0.20% sodium benzoate, and 0.40% phenethyl alcohol” had a bacteriostatic effect against Candida albicans, and the inhibition rates were 86.56% and 66.74% respectively. When the pH value was 4.4, the solution containing these ingredients had an inhibition rate of 37.61% against Candida albicans.
In summary, it could be seen that the lower the pH value of the solution containing “0.06% citric acid, 1.17% adipic acid, 0.2.0% propionic acid, 0.60% butyric acid, 0.20% sodium benzoate, and 0.40% phenethyl alcohol” was, the stronger the bacteriostatic effect of the solution against Candida albicans became.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.86. The experimental results are shown in Table 25.
Results:
1. It was evident in the result of Group 1 in Table 25 that, when the pH value was 3.86, the solution containing “1.46% adipic acid, 0.20% malic acid, 0.12% sodium benzoate, 0.20% propionic acid, 0.45% butyric acid, 0.30% phenethyl alcohol, and 1.80% palatinose” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 75.14%;
2. It was evident in the result of Group 2 in Table 25 that, when the pH value was 3.86, the solution containing “1.46% adipic acid, 0.20% malic acid, 0.12% sodium benzoate, 0.20% propionic acid, 0.45% butyric acid, 0.30% phenethyl alcohol, 1.80% palatinose, and 0.04% puerarin” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 78.45%;
3. It was evident in the result of Group 3 in Table 25 that, when the pH value was 3.86, the solution containing “1.46% adipic acid, 0.20% malic acid, 0.12% sodium benzoate, 0.20% propionic acid, 0.45% butyric acid, 0.30% phenethyl alcohol, 1.80% palatinose, and 0.06% puerarin” had a bacteriostatic effect against Candida albicans, and the inhibition rate was 76.24%.
In summary, it could be seen that when the pH value of the solution was 3.86, the 0.04% and 0.06% puerarin contained in the solution respectively did not affect the inhibitory effect of the solution against Candida albicans, and did not weaken the bacteriostatic effect.
Understandably, other estrogens such as diethylstilbestrol, hexoestrol, estradiol, estrone, estriol, nilestriol, ethinyloestradiol, quinestrol, mestranol, and promestriene, and other phytoestrogens such as daidzin, daidzein, glycitein, puerarin, coumestrol, genistein, equol, apigenin, genistin, genisteol, biochanin, coumestrol, formononetin, resveratrol, secoisolariciresinol, and lignan are similar to puerarin in property and action principle. Therefore, it can be inferred that said estrogens and phytoestrogens will not significantly weaken the inhibitory effect of the compositions against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Candida albicans ATCC 10231, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 4.0. The experimental results are shown in Table 26.
Results:
It was evident in the comparison of experimental results of Group 1 to Group 2 in Table 26 that, when the pH value was 4.0, 0.55% glutamic acid did not affect the bacteriostatic effect of the solution containing “0.14% malic acid, 1.46% adipic acid, 0.20% sodium benzoate, 1.67% propionic acid, and 0.40% phenethyl alcohol” against Candida albicans.
Understandably, other appropriate amino acids such as glutamine, L-aspartic acid, asparagine, leucine, isoleucine, phenylalanine, valine, proline, and threonine are similar to glutamic acid in property and action principle. Therefore, it can be inferred that said amino acids will not affect the inhibitory effect of the compositions against Candida albicans.
According to the method described in Appendix C of GB 15979-2002 Hygienic Standard for Disposable Sanitary Products, aqueous solutions containing different ingredients were studied for their bacteriostatic effects of 20 minutes of action against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 6538, the percentage concentration of each ingredient therein was the weight percentage concentration % (w/w), and the pH value of each group was 3.8. The experimental results are shown in Table 27.
Escherichia
Staphylococcus
coli
aureus
Results:
1. It was evident in the results of Groups 1-4 in Table 27 that, when the pH value was 3.8, the combinations of 1.75% adipic acid and each one of 1.50% propionic acid, 1.30% butyric acid, 0.50% valeric acid, or 0.20% hexanoic acid respectively had a strong bacteriostatic effect against Escherichia coli, and the inhibition rates were 100.00%, and had a bacteriostatic effect or a strong bacteriostatic effect against Staphylococcus aureus, and the inhibition rates were 50%-90% or >90%;
2. It was evident in the results of Groups 5-8 in Table 27 that, when the pH value was 3.8, the combinations of “0.03% cinnamyl alcohol and 1.75% adipic acid” and each one of 1.50% propionic acid, 1.30% butyric acid, 0.50% valeric acid, and 0.20% hexanoic acid respectively had a strong bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were 100.00%;
3. It was evident in the results of Groups 9-12 in Table 27 that, when the pH value was 3.8, the combinations of “0.03% cinnamic acid and 1.75% adipic acid” and each one of 1.50% propionic acid, 1.30% butyric acid, 0.50% valeric acid, or 0.20% hexanoic acid respectively had a strong bacteriostatic effect against Escherichia coli and Staphylococcus aureus, and the inhibition rates were 100.00%.
Basically by referring to the methods and results determination criteria in Volume IV 1121 Bacteriostatic Effectiveness Testing of the Pharmacopoeia of the People's Republic of China 2020, the bacteriostatic composition was studied for their bacteriostatic effects against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Aspergillus niger. The experimental results are shown in Table 28.
Staphylococcus
NI3
aureus
Pseudomonas
aeruginosa
Escherichia
coli
Candida
albicans
Aspergillus
niger
1The composition included: 1.50% (w/w) adipic acid, 0.14% (w/w) sodium benzoate, 0.32% (w/w) phenethyl alcohol, 0.54% (w/w) propionic acid, 0.20% (w/w) hexanoic acid, 0.20% (w/w) palatinose, 0.60% (w/w) maltose, 0.0005% (w/w) rose essential oil, and 2.15% (w/w) xanthan gum.
2The difference between the lg value of bacteria count measured at each interval and the lg value of the count of bacteria inoculated in 1 mL (g) of the test sample.
3No increase (NI) is defined as not more than 0.5 lg in the test bacteria count compared to the previous measurement time.
It was evident in the results in Table 28 that, when the pH value was 4.2, the reduced logarithm of the bacterial concentration was 5 on day 14 relative to day 0, and did not increase on day 28 relative to day 14 for all test bacteria, which met the requirements. It could be seen that the composition with a pH value of 4.2 in the test had an antiseptic effect.
It should be noted that even if the experimental conditions and methods are identical, the results of the in vitro bacteriostatic experiments, such as inhibition rate, may still differ due to the existence of experimental errors when the experiments are repeated, but such differences are reasonable and are understandable to those skilled in the art. Therefore, if there is any inconsistency between the in vitro experimental data in this disclosure and the data in the previous application documents, the data in this disclosure shall prevail.
In Vivo Experiment I
Three groups of gels containing different ingredients respectively were vaginally administered to Cynomolgus Monkeys, 0.5 mL once a day, for five consecutive days. Vaginal swabs were collected for the test of pH value and for smear staining and microscopic examination to observe the effects of the gels on the pH value of vaginal secretions and on the vaginal flora of Cynomolgus Monkeys. The experimental results are shown in Table 29:
1The gel of Group B included 2.50% (w/w) maltose, 0.20% (w/w) isomaltulose, 0.16% (w/w) sodium benzoate, 0.60% (w/w) sodium propionate, 0.35% (w/w) phenethyl alcohol, 1.00% (w/w) citric acid, 1.168% (w/w) adipic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.6.
2The gel of Group C included 2.50% (w/w) maltose, 0.20% (w/w) isomaltulose, 0.16% (w/w) sodium benzoate, 0.60% (w/w) sodium propionate, 0.35% (w/w) phenethyl alcohol, 0.61% (w/w) succinic acid, 1.168% (w/w) adipic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.6.
3The gel of Group 2 included 1.15% (w/w) sodium propionate, 0.45% (w/w) phenethyl alcohol, 0.20% (w/w) sodium benzoate, 1.50% (w/w) maltose, 1.00% (w/w) citric acid, and 2.15% (w/w) xanthan gum. with pH value adjusted to 3.6.
4Nugent score refers to the scoring based on the staining, morphology, and quantity of bacteria under microscopic examination of vaginal secretions after smearing and Gram staining, specified as follows: ≥7: Vaginal flora is mainly gram-negative bacilli, and/or negative cocci and positive cocci, with no or few gross gram-positive bacilli; 4-6: Vaginal flora is mainly gram-negative bacilli, and/or negative cocci and positive cocci, with few gross gram-positive bacilli; 0-3: Vaginal flora is normal, with mainly gross gram-positive bacilli.
Results:
1. It was evident in the results of Group B in Table 29 that, after five administrations of the gel that had a pH value of 3.6 and contained “2.50% (w/w) maltose, 0.20% (w/w) isomaltulose, 0.16% (w/w) sodium benzoate, 0.60% (w/w) sodium propionate, 0.35% (w/w) phenethyl alcohol, 1.00% (w/w) citric acid, 1.168% (w/w) adipic acid, and 2.15% (w/w) xanthan gum”, the pH values of the vaginal secretions decreased to 3.8 and the Nugent scores of the vaginal flora decreased to 0-3 in all the three Cynomolgus Monkeys;
2. It was evident in the results of Group C in Table 29 that, after five administrations of the gel that had a pH value of 3.6 and contained “2.50% (w/w) maltose, 0.20 (w/w) isomaltulose, 0.16% (w/w) sodium benzoate, 0.60% (w/w) sodium propionate, 0.35% (w/w) phenethyl alcohol, 0.61% (w/w) succinic acid, 1.168% (w/w) adipic acid, and 2.15% (w/w) xanthan gum”, the pH values of the vaginal secretions decreased to 3.8 in all the three Cynomolgus Monkeys, and the Nugent scores of the vaginal flora decreased to 0-3 in one Cynomolgus Monkey and to 4-6 in two Cynomolgus Monkeys;
3. It was evident in the results of Group 2 in Table 29 that, after five administrations of the gel that had a pH value of 3.6 and contained “1.15% (w/w) sodium propionate, 0.45% (w/w) phenethyl alcohol, 0.20% (w/w) sodium benzoate, 1.50% (w/w) maltose, 1.00% (w/w) citric acid, and 2.15% (w/w) xanthan gum”, the pH values of the vaginal secretions decreased to 3.8 in three of the five Cynomolgus Monkeys and to 4.4 in one Cynomolgus Monkey, and one terminated due to menstruation; and the Nugent scores of the vaginal flora decreased to 0-3 in three Cynomolgus Monkeys, one remained >7, and one terminated due to menstruation.
In summary, it could be seen that all the three groups of gels, which had a pH value of 3.6, had the efficacies of inhibiting abnormal vaginal flora, restoring vaginal acidity, and restoring vaginal lactobacilli. Between the gels of Groups B and C, which both contained adipic acid, and contained either citric acid or succinic acid respectively, Group B had better efficacy than Group C and Group 2, the latter did not contain adipic acid. The gel of Group 2 containing no adipic acid had the weakest efficacy in inhibiting abnormal vaginal flora, in restoring vaginal acidity, and in restoring vaginal lactobacilli among the three groups of gels.
In Vivo Experiment II
Three groups of gels containing different ingredients respectively were vaginally administered to Rhesus Monkeys, 0.5 ml once a day, for five consecutive days. Vaginal secretions were collected for the test of pH value and for smear staining and microscopic examination, to observe the effects of the gels on the pH values of vaginal secretions and on the vaginal flora of Rhesus Monkeys. The experimental results are shown in Table 30:
1The gel of Group A included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.60% (w/w) butyric acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 4.1.
2The gel of Group B included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium
3The gel of Group C included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.60% (w/w) butyric acid, 0.0015% (w/w) capric acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 4.1.
4Same as Note 4 in in vivo experiment I.
Results:
1. It was evident in the results of Group A that, after five administrations of the gel, which had a pH value of 4.1 and contained “1.31% (w/w) adipic acid, 0.35% (w/w) malic acid, 0.30% (w/w) phenethyl alcohol, 0.12% (w/w) sodium benzoate, and 0.60% (w/w) butyric acid”, as well as 2.00% (w/w) isomaltulose and 2.15% (w/w) xanthan gum, the pH values of vaginal secretions decreased to 3.8 in three of the four Rhesus Monkeys, while their Nugent scores of the vaginal flora decreased to 0-3; the pH value of vaginal secretion remained 5.4 in one Rhesus Monkey, and the Nugent score of vaginal flora decreased to 4-6.
2. It was evident in the results of Group B that, after five administrations of the gel, which had a pH value of 4.1 and contained “1.31% (w/w) adipic acid, 0.35% (w/w) malic acid, 0.30% (w/w) phenethyl alcohol, 0.12% (w/w) sodium benzoate, 0.60% (w/w) butyric acid, and 0.0010% (w/w) capric acid”, as well as 2.00% (w/w) isomaltulose and 2.15% (w/w) xanthan gum, the pH values of vaginal secretions decreased to 3.8 in three of the five Rhesus Monkeys, while their Nugent scores of the vaginal flora decreased to 0-3; the pH value decreased to 4.1 in one of the rest, while the Nugent score decreased to 4-6; and the pH value remained 5.4 in another one of the rest, while the Nugent score decreased to 4-6.
3. It was evident in the results of Group C that, after five administrations of the gel, which had a pH value of 4.1 and contained “1.31% (w/w) adipic acid, 0.35% (w/w) malic acid, 0.30% (w/w) phenethyl alcohol, 0.12% (w/w) sodium benzoate, 0.60% (w/w) butyric acid, and 0.0015% (w/w) capric acid”, as well as 2.0% (w/w) isomaltulose and 2.15% (w/w) xanthan gum, the pH values of vaginal secretions decreased to 3.8 in one of the two Rhesus Monkeys, while the Nugent score of the vaginal flora decreased to 0-3; and the pH value in another Rhesus Monkeys decreased to 4.1, while the Nugent score decreased to 4-6.
In summary, it could be seen that the three groups of gels containing 0.60% (w/w) butyric acid, 0.00125% (w/w) capric acid, or 0.0015% (w/w) capric acid respectively in the experiment had the effect of inhibiting abnormal vaginal flora, modulating vaginal flora, restoring vaginal lactobacilli, and restoring vaginal acidity to normal.
In Vivo Experiment III
Five groups of gels containing different ingredients respectively were vaginally administered to Cynomolgus Monkeys, 0.5 mL once a day, for five consecutive days. Vaginal swabs were collected for the test of pH value and for smear staining and microscopic examination to observe the effects of the gels on the pH values of vaginal secretions and on the vaginal flora of Cynomolgus Monkeys. The experimental results are shown in Table 31:
1The gel of Group 1 included 0.14% (w/w) sodium benzoate, 0.54% (w/w) propionic acid, 1.46% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
2The gel of Group 2 included 2.00% (w/w) maltose, 0.14% (w/w) sodium benzoate, 0.54% (w/w) propionic acid, 1.46% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
3The gel of Group 3 included 2.0% (w/w) isomaltulose, 0.14% (w/w) sodium benzoate, 0.54% (w/w) propionic acid, 1.46% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
4The gel of Group 4 included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.0015% (w/w) capric acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.9.
5The gel of Group 5 included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.01% (w/w) caprylic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.9.
6Same as Note 4 in in vivo experiment I.
Results:
1. It was evident in the results of Groups 1-3 in Table 31 that all of the ingredients except saccharide of the gels of the three groups are identical. The gel of Group 1 did not contain any saccharide, the gel of Group 2 contained 2.00% (w/w) maltose, and the gel of Group 3 contained 2.00% (w/w) isomaltulose. The gels had the same pH value of 3.8, and all contained “0.14% (w/w) sodium benzoate, 0.54% (w/w) propionic acid, 1.46% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum”.
After five administrations, the pH values of vaginal secretions decreased to 4.1 or below in all three groups; the Nugent scores of the vaginal flora in Group 1 without saccharide decreased to 4-6 in two of the four Cynomolgus Monkeys, and to 0-3 in the other two Cynomolgus Monkeys; The Nugent scores in Group 2 with maltose decreased to 0-3 in two Cynomolgus Monkeys. The Nugent scores in Group 3 with isomaltulose decreased to 0-3 in two of the three Cynomolgus Monkeys and to 4-6 in the other one Cynomolgus Monkey.
2. It was evident in the results of Group 4 in Table 31 that, after five administrations of the gel, which had a pH value of 3.9 and contained “2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, 0.0015% (w/w) capric acid, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 3.8 in two Cynomolgus Monkeys, and the Nugent score of the vaginal flora decreased to 0-3 in one Cynomolgus Monkey and to 4-6 in another Cynomolgus Monkey.
3. It was evident in the results of Group 5 in Table 31 that, after three administrations of the gel, which had a pH value of 3.9 and contained “2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, 0.01% (w/w) caprylic acid, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 3.8 and the Nugent score of the vaginal flora decreased to 0-3 in two Cynomolgus Monkeys.
In summary, it could be seen that:
1. The gel which had a pH value of 3.8 and contained “1.46% (w/w) adipic acid, 0.35% (w/w) malic acid, 0.54% (w/w) propionic acid, 0.30% (w/w) phenethyl alcohol, and 0.14% (w/w) sodium benzoate” but no saccharide had the efficacy of inhibiting abnormal vaginal flora, restoring vaginal acidity, and restoring vaginal lactobacilli, as shown in the results of Group 1. However, the gel of Group 2 which contains maltose further and the gel of Group 3 which contains isomaltulose further had a faster and stronger effect in restoring vaginal lactobacilli than the gel of Group 1 without saccharide, as shown in the results of Group 2 and Group 3.
2. The gels which contained “1.31% (w/w) adipic acid, 0.35% (w/w) malic acid, 0.38% (w/w) propionic acid, 0.30% (w/w) phenethyl alcohol, 0.12% (w/w) sodium benzoate, and 2.0% (w/w) isomaltulose”, as well as 0.0015% (w/w) capric acid or 0.01% (w/w) caprylic acid respectively had the efficacy of inhibiting abnormal vaginal flora, restoring the vaginal acidity, and restoring vaginal lactobacilli, as shown in the results of Group 4 and Group 5.
In Vivo Experiment IV
Three groups of gels that contained different ingredients were vaginally administered to Cynomolgus Monkeys, 0.5 ml once a day, for five consecutive days. Vaginal swabs were collected for the test of pH value, and for smear staining and microscopic examination to observe the effects of the gels on the pH values of vaginal secretions and on the vaginal flora of Cynomolgus Monkeys. The experimental results are shown in Table 32:
1The gel of Group 1 included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.04% (w/w) heptanoic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.9.
2The gel of Group 2 included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.10% (w/w) hexanoic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.9.
3The gel of Group 3 included 2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.20% (w/w) valeric acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.9.
4Same as Note 4 in in vivo experiment I
Results:
1. It was evident in the result of Group 1 in Table 32 that, after five administrations of the gel, which had a pH value of 3.9 and contained “2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, 0.04% (w/w) heptanoic acid, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretion decreased to 3.8 and the Nugent score of the vaginal flora decreased to 0-3 in Cynomolgus Monkeys.
2. It was evident in the results of Group 2 in Table 32 that, after five administrations of the gel, which had a pH value of 3.9 and contained “2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, 0.10% (w/w) hexanoic acid, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 4.1 or below, and the Nugent scores of the vaginal flora decreased to 0-3 in the two Cynomolgus Monkeys.
3. It was evident in the results of Group 3 in Table 32 that, after five administrations of the gel, which had a pH value of 3.9 and contained “2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, 0.20% (w/w) valeric acid, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions remained 5.4 in the two Cynomolgus Monkeys, and the Nugent scores of the vaginal flora decreased to 0-3 in one Cynomolgus Monkey and to 4-6 in another Cynomolgus Monkey.
In summary, it could be seen that the three groups of gels, which had a pH value of 3.9 and contained “2.00% (w/w) isomaltulose, 0.12% (w/w) sodium benzoate, 0.38% (w/w) propionic acid, 1.31% (w/w) adipic acid, 0.30% (w/w) phenethyl alcohol, 0.35% (w/w) malic acid, and 2.15% (w/w) xanthan gum”, as well as 0.04% (w/w) heptanoic acid, 0.10% (w/w) hexanoic acid, or 0.20% (w/w) valeric acid respectively had the efficacy of inhibiting abnormal vaginal flora and restoring vaginal lactobacilli. The two groups of gels containing 0.04% (w/w) heptanoic acid or 0.10% (w/w) hexanoic acid respectively also had the efficacy of restoring vaginal acidity.
In Vivo Experiment V
Six groups of gels containing different ingredients were vaginally administered to Cynomolgus Monkeys, 0.5 mL once a day, for five consecutive days. The vaginal swabs were collected for the test of pH values and for smear staining and microscopic examination to observe the effects of the gels on the pH values of vaginal secretions and on the vaginal flora of Cynomolgus Monkeys. The experimental results are shown in Table 33:
1The gel of Group 1 included 0.10% (w/w) hexanoic acid, 1.50% (w/w) adipic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.1.
2The gel of Group 2 included 2.00% (w/w) propionic acid, 0.30% (w/w) cinnamyl alcohol, 0.60% (w/w) succinic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
3The gel of Group 3 included 0.02% (w/w) caprylic acid, 0.25% (w/w) benzoic acid, 1.00% (w/w) citric acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
4The gel of Group 4 included 0.20% (w/w) benzoic acid, 0.25% (w/w) benzyl alcohol, 0.001% (w/w) undecylic acid, 1.00% (w/w) malic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.80.
5The gel of Group 5 included 0.20% (w/w) benzoic acid, 0.50% (w/w) propionic acid, 1.50% (w/w) adipic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
6The gel of Group 6 included 1.00% (w/w) pimelic acid, 0.60% (w/w) phenethyl alcohol, 1.25% (w/w) butyric acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
7Same as Note 4 in in vivo experiment I.
Results:
1. It was evident in the results of Group 1 in Table 33 that, after five administrations of the gel, which had a pH value of 3.1 and contained “0.10% (w/w) hexanoic acid, 1.50% (w/w) adipic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the experiment was terminated in one of the three Cynomolgus Monkeys due to menstruation; the pH values of vaginal secretions remained 5.4 in one Cynomolgus Monkey, and decreased to 3.8 in another one; The Nugent scores of the vaginal flora decreased to 4-6 in one Cynomolgus Monkey and decreased to 0-3 in another one;
2. It was evident in the results of Group 2 in Table 33 that, after five administrations of the gels, which had a pH value of 3.8 and contained “2.00% (w/w) propionic acid, 0.30% (w/w) cinnamyl alcohol, 0.60% (w/w) succinic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 4.1 in one of the two Cynomolgus Monkeys and remained 5.4 in another one; The Nugent scores of the vaginal flora decreased to 4-6 in one Cynomolgus Monkey and to 0-3 in another one.
3. It was evident in the results of Group 3 in Table 33 that, after five administrations of the gel, which had a pH value of 3.8 and contained “0.02% (w/w) caprylic acid, 0.25% (w/w) benzoic acid, 1.00% (w/w) citric acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 3.8 in one of the two Cynomolgus Monkeys and to 4.1 in another, and the Nugent scores of the vaginal flora decreased to 0-3 in the two Cynomolgus Monkeys;
4. It was evident in the results of Group 4 in Table 33 that, after five administrations of the gel, which had a pH value of 3.8 and contained“0.20% (w/w) benzoic acid, 0.25% (w/w) benzyl alcohol, 0.001% (w/w) undecylic acid, 1.00% (w/w) malic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 4.6 in one of the two Cynomolgus Monkeys and to 3.8 in another, and the Nugent score of the vaginal flora decreased to 4-6 in one Cynomolgus Monkey and to 0-3 in another;
5. It was evident in the results of Group 5 in Table 33 that, after five administrations of the gel, which had a pH value of 3.8 and contained “0.20% (w/w) benzoic acid, 0.50% (w/w) propionic acid, 1.50% (w/w) adipic acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH value of vaginal secretions decreased to 4.6 in one of the two Cynomolgus Monkeys and to 3.8 in another, and the Nugent score of vaginal flora decreased to 4-6 in one Cynomolgus Monkey and to 0-3 in another;
6. It was evident in the results of Group 6 in Table 33 that, after five administrations of the gel, which had a pH value of 3.8 and contained “1.00% (w/w) pimelic acid, 0.60% (w/w) phenethyl alcohol, 1.25% (w/w) butyric acid, 2.00% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH value of vaginal secretions decreased to 3.8 in one of the two Cynomolgus Monkeys and to 4.1 in another, and the Nugent score of the vaginal flora decreased to 0-3 in one Cynomolgus Monkey and to 4-6 in another.
In summary, it could be seen that these 6 gel compositions had the effect of inhibiting abnormal vaginal flora, restoring vaginal acidity, and restoring vaginal lactobacilli.
In Vivo Experiment VI
The gels containing adipic acid or fumaric acid were vaginally administered to Cynomolgus Monkeys, 0.5 mL once a day, for five consecutive days, and vaginal swabs were collected for the test of pH value and for smear staining and microscopic examination to observe the effects of the gels on the pH values of vaginal secretions and on the vaginal flora of Cynomolgus Monkeys. The experimental results are shown in Table 34:
1The gel of Group 1 included 1.50% (w/w) fumaric acid, 1.00% (w/w) propionic acid, 0.08% (w/w) cinnamic acid, 0.25% (w/w) phenethyl alcohol, 1.40% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
2The gel of Group 2 included1.50% (w/w) adipic acid, 1.00% (w/w) propionic acid, 0.08% (w/w) cinnamic acid, 0.25% (w/w) phenethyl alcohol, 1.40% (w/w) maltose, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.8.
3Same as Note 4 in in vivo experiment I.
Results:
1. It was evident in the results of Group 1 in Table 34 that, after five administrations of the gel, which had a pH value of 3.8 and contained “1.50% (w/w) fumaric acid, 1.00% (w/w) propionic acid, 0.08% (w/w) cinnamic acid, 0.25% (w/w) phenethyl alcohol, 1.40% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions remained 5.4 in two Cynomolgus Monkeys, and the Nugent score of the vaginal flora remained >7 in one Cynomolgus Monkey and decreased to 4-6 in another.
2. It was evident from the results of Group 2 in Table 34 that, after five administrations of the gel, which had a pH value of 3.8 and contained “1.50% (w/w) adipic acid, 1.00% (w/w) propionic acid, 0.08% (w/w) cinnamic acid, 0.25% (w/w) phenethyl alcohol, 1.40% (w/w) maltose, and 2.15% (w/w) xanthan gum”, the pH values of vaginal secretions decreased to 4.1 and the Nugent scores of the vaginal flora decreased to 0-3 in both of the two Cynomolgus Monkeys.
In summary, it could be seen that the gel containing 1.50% (w/w) adipic acid had the effect of inhibiting abnormal vaginal flora, restoring vaginal lactobacilli, and restoring vaginal acidity.
Clinical Observation I
The gel A was vaginally administered in 10 patients with bacterial vaginosis (BV) and 5 healthy volunteers, 4.5 g once a day, for 5 consecutive days, and vaginal swabs were collected on the third day of administration (V1) and 3 days after drug discontinuance (V2) for the test of pH value and for smear staining and microscopic examination to observe the effects of the gel on the pH values of vaginal secretions and on the vaginal flora. The experimental results are shown in Table 35:
1Gel A included 1.46% (w/w) adipic acid, 0.70% (w/w) sodium propionate, 0.14% (w/w) sodium benzoate, 0.32% (w/w) phenethyl alcohol, 1.00% (w/w) citric acid, 1.60% (w/w) maltose, 0.20% (w/w) palatinose, 0.0005% (w/w) rose essential oil, and 2.15% (w/w) xanthan gum, with pH value adjusted to 3.70.
2pH value was measured using a pH range standard colorimetric card. The colorimetric card has two ranges: 3.8-4.1 indicates normal; 4.6-5.4 indicates abnormal.
3Same as Note 7 in in vivo experiment I.
Results:
1. It was evident in the results of BV patients in Table 35 that, after three administrations of the gel, which had a pH value of 3.70 and contained “1.46% (w/w) adipic acid, 0.70% (w/w) sodium propionate, 0.14% (w/w) sodium benzoate, 0.32% (w/w) phenethyl alcohol, 1.00% (w/w) citric acid, 1.60% (w/w) maltose, 0.20% (w/w) palatinose, 0.0005% (w/w) rose essential oil, and 2.15% (w/w) xanthan gum”, the pH values decreased to 3.8-4.1 in seven of the ten BV patients, and the Nugent scores of the vaginal flora decreased to 3 or below in three BV patients and to 4-6 in four BV patients.
After five administrations and an interval of 3 days after drug discontinuance, the pH values decreased to 3.8-4.1 in nine of the ten BV patients, and the Nugent scores of the vaginal flora decreased to lower than 3. In one patient, the pH value remained 4.6-5.4, and the Nugent score decreased to 6.
2. It was evident in the results of healthy volunteers in Table 35 that, three administrations of the gel, which had a pH value of 3.70 and contained “1.46% (w/w) adipic acid, 0.70% (w/w) sodium propionate, 0.14% (w/w) sodium benzoate, 0.32% (w/w) phenethyl alcohol, 1.00% (w/w) citric acid, 1.60% (w/w) maltose, 0.20% (w/w) palatinose, 0.0005% (w/w) rose essential oil, and 2.15% (w/w) xanthan gum”, did not affect the pH values of vaginal secretions or the vaginal flora in healthy volunteers.
After five administrations and an interval of 3 days after drug discontinuance, the results showed that the administrations did not affect the pH value of vaginal secretions or the vaginal flora in healthy volunteers.
In summary, it could be seen in the experiment that, the gel, which had a pH value of 3.70 and contained“1.46% (w/w) adipic acid, 0.70% (w/w) sodium propionate, 0.14% (w/w) sodium benzoate, 0.32% (w/w) phenethyl alcohol, 1.00% (w/w) citric acid, 1.60% (w/w) maltose, 0.20% (w/w) palatinose, 0.0005% (w/w) rose essential oil, and 2.15% (w/w) xanthan gum”, could decrease significantly the abnormal vaginal flora, increase significantly vaginal lactobacilli, and lowered the vaginal pH value in BV patients, but had no effect on the normal pH values of vaginal secretions or normal flora in healthy volunteers, indicating that the gel had the effect of inhibiting abnormal vaginal flora, restoring and/or maintaining vaginal lactobacilli, and restoring and/or maintaining normal vaginal acidity.
Although the present invention has been described by referring to exemplary embodiments, it should be understood that the present invention is not limited to the disclosed exemplary embodiments. Various adjustments or variations can be made to the exemplary embodiments of the specification without departing from the scope or spirit of the invention. The scope of the claims is to be based on the broadest interpretation to cover all modifications, equivalent structures, and functions.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202010101402.6 | Feb 2020 | CN | national |
| 202011324292.6 | Nov 2020 | CN | national |
The present application is a continuation of PCT application No. PCT/CN2021/075655, filed on Feb. 5, 2021, which claims the priority of China Patent Application No. 202010101402.6, filed on Feb. 12, 2020, and China Patent Application No. 202011324292.6, filed on Nov. 15, 2020. The entirety of each of the above mentioned patent applications is incorporated by reference herein and made a part of this specification.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/CN2021/075655 | Feb 2021 | US |
| Child | 17886476 | US |
