KR20070057884A - Non-absorbent and absorbent articles for inhibiting the production of exoproteins - Google Patents

Abstract

Non-absorbent and absorbent articles for inhibiting the production of exoproteins from Gram positive bacteria are disclosed. The non-absorbent and absorbent articles include an effective amount of a precursor compound having the general formula (I): wherein R1 is selected from the group consisting of formula (II) and formula (III); R7 is -OCH2-; X is 0 or 1; R5 is a substituted or unsubstituted aromatic ring or a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl moiety that may or may not be substituted with hetero atoms; R6 is selected from the group consisting of an amino acid, a methyl ester of an amino acid, and an ethyl ester of an amino acid; R2, R3, and R4 are independently selected from the group consisting of H, OH, COOH.

Description

NON-ABSORBENT AND ABSORBENT ARTICLES FOR INHIBITING THE PRODUCTION OF EXOPROTEINS}

The present invention relates to the inhibition of exocrine protein production in or around the vagina of a woman associated with a non-absorbable or absorbent article. More particularly, the present invention incorporates a precursor compound into a non-absorbent or absorbent article or on a non-absorbent or absorbent article, so that, in use, the precursor compound is hydrolyzed by enzymatic activity in or around the vagina to It is directed to allowing active species to be produced which reduces exocrine protein production.

Disposable absorbent articles, such as vaginal tampons, for the absorption of vaginal excretion are widely used. Such disposable articles typically have a compacted mass of absorbent shaped into the desired form, typically as dictated by the intended consumer use. In the field of vaginal tampons, the instrument is intended to be inserted into the vagina for the absorption of body fluids that are usually drained during the female menstrual period.

There is a complex process in the woman's body that keeps the vaginal and physiologically relevant areas healthy. In women between menarche and menopause, normal vagina provides an ecosystem for a variety of microorganisms. Bacteria are the predominant type of microorganisms present in the vagina, and most women inhabit about 10 ⁹ bacteria per gram of vaginal fluid. The bacterial flora of vagina consists of both aerobic and anaerobic bacteria. More commonly isolated bacteria include Lactobacillus spp., Corynebacteria spp., Gardnerella spp. vaginalis), Staphylococcus aureus (Staphylococcus) species, peptidase Toko kusu (Peptococcus) species, and aerobic and anaerobic Streptococcus (Streptococcus) species, and peeling teroyi des (Bacteroides) species. Sometimes with other microorganisms isolated from vaginal yeast (Candida albicans (Candida albicans )), protozoa ( Trichomonas baginalis) vaginalis)), mycoplasma (Mycoplasma hoe Nice (Mycoplasma hominis)), chlamydia (Chlamydia trachomatis (Chlamydia trachomatis)), and viral (herpes simplex (Herpes simplex )). These latter organisms are generally associated with vaginitis or sexually transmitted diseases, but they can be present in small numbers without causing symptoms.

Physiological, social and idiosyncratic factors affect the amount and species of bacteria present in the vagina. Physiological factors include age, menstrual cycle date, and pregnancy. For example, microorganisms present in the vagina throughout the menstrual cycle may include Lactobacillus, Corynebacterium, Urea Plasma and Mycoplasma. The number of microorganisms and the types of microorganisms are unique to each individual. Social and specific constitution factors include birth control methods, sexual behavior, systemic diseases (eg, diabetes), and medication.

Bacterial proteins and metabolites produced in the vagina can affect other microorganisms and human hosts. For example, the vagina between menstrual cycles is moderately acidic with a pH ranging from about 3.8 to about 4.5. This pH range is generally regarded as the most favorable condition for the maintenance of a normal flora. At this pH, microorganisms in many species of balanced ecology live in the vagina. These microorganisms play a beneficial role in providing protection and resistance to infections and keep some species of bacteria, such as Staphylococcus aureus , from retaining in the vagina. Low pH is a result of the growth of lactobacillus and the production of their acidic products. Microorganisms in the vagina can also produce antimicrobial compounds such as fungicides and hydrogen peroxide for other bacterial species. One example is lactocin, a bacteriocin-like product of Lactobacillus against other Lactobacillus species.

Some microbial products produced in the vagina can negatively affect human hosts. For example, Staphylococcus aureus can be a variety of enzymes, including enterotoxins, Toxic Shock Syndrome Toxin-1 (TSST-1), and enzymes such as esterases and amidases. It can produce and secrete exocrine proteins into its environment. When absorbed into the bloodstream of a host, TSST-1 can cause toxic shock syndrome (TSS) in non-immune humans.

Staphylococcus aureus is found in the vagina of about 16% of healthy women of menstrual age. Not all strains of Staphylococcus aureus can produce TSST-1. About 25% of these women will live with Staphylococcus aureus, which produces TSST-1. TSST-1 and some staphylococcal enterotoxin have been shown to cause TSS in humans.

Symptoms of toxic shock syndrome generally include fever, diarrhea, vomiting and rash followed by a sudden drop in blood pressure. In about 6% of people with disease, multiple organ failure occurs. Staphylococcus aureus does not initiate toxic shock syndrome as a result of the invasion of microorganisms into the vaginal cavity. As Staphylococcus aureus grows and proliferates, it can produce TSST-1. Only after entering the bloodstream, TSST-1 acts systemically, causing symptoms due to toxic shock syndrome.

The pH of menstrual fluid is about 7.3. During menstruation, the pH of the vagina migrates toward neutral and can become slightly alkaline. This change causes microorganisms to proliferate whose growth is inhibited by the acidic environment. For example, Staphylococcus aureus is more frequently isolated from vaginal swabs collected during menstruation than from vaginal swabs collected between menstrual cycles.

When Staphylococcus aureus is present in the realm of the human body in which normal microbial populations, such as vaginas, it can be difficult to eradicate Staphylococcus aureus without harming the members of the normal microflora required for a healthy ecosystem. Typically, antibiotics that kill Staphylococcus aureus are not selected for use in products that are inserted into the vagina due to their effect on normal vaginal microflora. An alternative to complete eradication is a technique designed to prevent or substantially reduce the ability of bacteria to produce toxins.

Numerous attempts have been made to reduce or eliminate toxic shock syndrome caused by pathogenic microorganisms and menstruation by incorporating one or more bacteriostatic, bactericidal and / or detoxifying compounds into the vaginal product. For example, L-ascorbic acid was applied to menstrual tampons to detoxify toxins found in the vagina. Others have incorporated monoesters and diesters of polyhydric aliphatic alcohols such as glycerol monolaurate as bactericidal compounds (see, eg, US Pat. No. 5,679,369). Others have introduced other nonionic surfactants such as alkyl ethers, alkyl amines and alkyl amides as detoxification compounds (see, eg, US Pat. Nos. 5,685,872, 5,618,554 and 5,612,045).

One important problem associated with some of the existing attempts above is that the compounds used can be very volatile during incorporation into non-absorbent or absorbent articles and during further fabrication processes. In particular, it has been found that compounds such as some aromatic compounds, terpenes, and isoprenoids are completely volatilized from the non-absorbent or absorbent product during the high temperature fabrication step. In addition, some compounds may have volatility problems during storage prior to use by the consumer.

Thus, personal hygiene products such as non-absorbing or absorbing products that contain inhibitory compounds that effectively inhibit the production of endocrine proteins such as TSST-1 from Gram-positive bacteria without being substantially harmful to the natural flora found in the vaginal region. This is constantly required. In addition, such inhibitory compounds need to remain active even in the presence of enzymes such as lipases, esterases and amidases that can adversely affect efficacy and may also be present in the vagina. In order to deliver effective inhibitors to the consumer, it is desirable for the compound to have low volatility and remain in the product during manufacture, storage and transportation.

Summary of the Invention

The present invention relates to non-absorbable products and absorbent articles that inhibit the production of exocrine proteins from Gram-positive bacteria. More particularly, the present invention relates to vaginal tampons or non-absorbing substrates, such as tampon applicators, incorporating at least one precursor compound formed by linking at least one aromatic compound with at least one second compound via an ester or amide bond. will be. Once introduced into the vagina, these precursor compounds can be hydrolyzed by enzymes produced by the natural vaginal flora, inhibiting the production of exocrine proteins from Gram-positive bacteria without substantially affecting the flora in the vagina. Active species can be produced. In some embodiments, the precursor compound itself can also inhibit the production of exocrine proteins from Gram positive bacteria.

Thus, the present invention relates to an exocrine protein inhibitor for inhibiting the production of exocrine proteins from Gram-positive bacteria. Exocrine protein inhibitors include non-absorbing substrates suitable for incorporation into the vagina, with an effective amount of a precursor compound of the formula having been deposited thereon, which inhibits the production of endocrine proteins from Gram-positive bacteria upon hydrolysis. Effective active species can be produced:

및

으로 구성되는 군으로부터 선택되고; R⁷은 -OCH₂-이고; X는 0 또는 1이고; R⁵는 치환 또는 비치환 방향족 고리, 또는 헤테로 원자로 치환되거나 치환되지 않을 수 있는, 1가의 포화 또는 불포화, 치환 또는 비치환, 분지쇄 또는 직쇄 히드로카르빌 잔기이고; R⁶은 아미노산, 아미노산의 메틸 에스테르, 및 아미노산의 에틸 에스테르로 구성되는 군으로부터 선택되고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is

And

It is selected from the group consisting of; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

The present invention also relates to tampon applicators for inhibiting the production of exocrine proteins from Gram-positive bacteria. Tampon applicators comprise a non-absorbing material suitable for insertion into the vagina and in which an effective amount of a precursor compound of the formula is deposited, wherein the precursor compound is effective to inhibit the production of exocrine proteins from Gram-positive bacteria upon hydrolysis. Active species can be produced:

으로 구성되는 군으로부터 선택되고; R⁷은 -OCH₂-이고; X는 0 또는 1이고; R⁵는 치환 또는 비치환 방향족 고리, 또는 헤테로 원자로 치환되거나 치환되지 않을 수 있는, 1가의 포화 또는 불포화, 치환 또는 비치환, 분지쇄 또는 직쇄 히드로카르빌 잔기이고; R⁶은 아미노산, 아미노산의 메틸 에스테르, 및 아미노산의 에틸 에스테르로 구성되는 군으로부터 선택되고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is And

It is selected from the group consisting of; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

The present invention also relates to an irrigation formulation for inhibiting the production of exocrine proteins from Gram-positive bacteria located in and around the vagina of women. Irrigation bath formulations include vaginal cleansing formulations comprising a pharmaceutically acceptable carrier and an effective amount of a precursor compound of the formula wherein the precursor compound is effective to inhibit the production of exocrine proteins from Gram-positive bacteria upon hydrolysis. Active species can be produced and vaginal cleansing formulations are suitable for use in the vagina of women:

및

으로 구성되는 군으로부터 선택되고; R⁷은 -OCH₂-이 고; X는 0 또는 1이고; R⁵는 치환 또는 비치환 방향족 고리, 또는 헤테로 원자로 치환되거나 치환되지 않을 수 있는, 1가의 포화 또는 불포화, 치환 또는 비치환, 분지쇄 또는 직쇄 히드로카르빌 잔기이고; R⁶은 아미노산, 아미노산의 메틸 에스테르, 및 아미노산의 에틸 에스테르로 구성되는 군으로부터 선택되고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is

And

It is selected from the group consisting of; R ⁷ is —OCH ₂ —; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

The invention also relates to absorbent articles for inhibiting the production of exocrine proteins from Gram-positive bacteria. Absorbent articles are suitable for insertion into the vagina and include an absorbent construct and an effective amount of a precursor compound of the formula wherein the precursor compound is capable of producing an active species that is effective at inhibiting the production of exocrine proteins from Gram-positive bacteria upon hydrolysis. have:

및

으로 구성되는 군으로부터 선택되고; R⁷은 -OCH₂-이고; X는 0 또는 1이고; R⁵는 치환 또는 비치환 방향족 고리, 또는 헤테로 원자로 치환되거나 치환되지 않을 수 있는, 1가의 포화 또는 불포화, 치환 또는 비치환, 분지쇄 또는 직쇄 히드로카르빌 잔기이고; R⁶은 아미노산, 아미노산의 메틸 에스테르, 및 아미노산의 에틸 에스테르로 구성되는 군으로부터 선택되고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is

And

It is selected from the group consisting of; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

The present invention also relates to vaginal tampons for inhibiting the production of exocrine proteins from Gram-positive bacteria. Vaginal tampons comprise an absorbent tampon material and an effective amount of a precursor compound of the formula: wherein the precursor compound can produce an active species that is effective at inhibiting the production of exocrine proteins from Gram-positive bacteria upon hydrolysis:

및

으로 구성되는 군으로부터 선택되고; R⁷은 -OCH₂-이고; X는 0 또는 1이고; R⁵는 치환 또는 비치환 방향족 고리, 또는 헤테로 원자로 치환되거나 치환되지 않을 수 있는, 1가의 포화 또는 불포화, 치환 또는 비치환, 분지쇄 또는 직쇄 히드로카르빌 잔기이고; R⁶은 아미노산, 아미노산의 메틸 에스테르, 및 아미노산의 에틸 에스테르로 구성되는 군으로부터 선택되고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is

And

It is selected from the group consisting of; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

Other features and advantages of the invention will be in part apparent and in part pointed out hereinafter.

In general, the present invention relates to non-absorbing or absorbent articles comprising precursor compounds from which active species are produced that can inhibit the production of exocrine proteins from Gram-positive bacteria upon hydrolysis. In particular, the present invention provides a ratio comprising a precursor compound formed by linking an aromatic compound to a second compound by ester or amide bonds, which can be readily hydrolyzed by enzymatic action within the vagina so that the active species and the second compound are produced. It relates to an absorbent product or an absorbent article. The active species has been found to substantially inhibit the production of exocrine proteins such as TSST-1 from Gram-positive bacteria. Typically, the precursor compound itself can also inhibit the production of exocrine proteins from Gram-positive bacteria. Additionally, precursor compounds can be used in combination with surfactants such as, for example, mireth-3 myristate, glycerol monolaurate, and / or laureth-4 to substantially inhibit the production of exocrine proteins from Gram-positive bacteria. can do.

While the present invention will be described in detail with reference to tampon applicators, those skilled in the art will appreciate that other non-absorbent articles, instruments, and / or products, such as non-absorbent incontinence instruments, barrier birth control instruments, non-absorbable contraceptive instruments, and tubes It will be appreciated that the present invention is applicable to a bathing apparatus.

As used herein, the idiom “non-absorbent article” generally refers to a substrate or apparatus that includes an outer layer formed from a substantially hydrophobic material that repels fluid, such as menses, blood products, and the like. Suitable materials for the construction of the non-absorbent articles of the present invention include, for example, rubber, plastic and cardboard.

Typically, the tampon applicator comprises an outer tube, which is preferably in the form of a hollow tube. The tube is formed from paper, cardboard, cardboard, plastic, thermoplastic film, aqueous coating or combinations thereof. If paper, cardboard or cardboard is used, it may be coated with wax or water-insoluble polymer which makes it waterproof. Suitable plastic materials include polyolefins such as low density polyethylene and low density polypropylene. The outer tube must have sufficient strength and rigidity to prevent collapse under normal vaginal pressure. The outer tube can also be shaped into a cylindrical shape with longitudinal seams, or wound spirally or in a circular shape. The diameter of the outer tube is relatively small, from about 10 mm to about 20 mm.

The outer tube has a spaced first end and a second end. The outer tube is formed from two or more separate layers, which can be built with the same or different board weights. The layers may be made from different materials, for example cardboard and film, or may be made from similar materials of different properties, for example board weights. In one embodiment, the outer layer can be formed from about 0.06 mm of coated thin cardboard or from a film material of about 0.01 mm in thickness, while the one or more inner layers are from uncoated material of higher board weight. Can be formed. The outer layer may consist of a high gloss coated paper that is water-degradable or water-dispersible. Alternatively, the outer layer can have different finishes, such as semi-gloss or satin finishes. The coating on the outer tube can be selected from a variety of materials. Suitable coatings include polyethylene, polypropylene, polyvinylidene chloride and polychloride alcohols. The outer layer can also be lubricated or contain additives. Suitable lubricants and additives include any pharmaceutically acceptable lubricant or additive commonly used in tampon applicators. Such lubricants and additives include organic compounds, long chain aliphatic groups such as derivatives of fatty acids such as stearamide and oleamide.

The paper used to construct the tampon applicator has a board weight / layer between about 20 pounds and about 200 pounds per year, suitably between about 25 pounds and about 100 pounds per year, more suitably about 30 per year. Should be pounds to about 50 pounds. "Lead" is defined as a material having dimensions 24 inches (609.6 mm) x 36 inches (914.4 mm) x 500 sheets. The thickness of each cardboard layer should be less than about 0.4 mm, suitably about 0.04 mm to about 0.2 mm, more suitably about 0.05 mm to about 0.16 mm. Typically, the outer layer will be thinner than the inner cardboard layer (s).

If one of the layers is made from a thermoplastic film, it may be polyethylene. Suitable polyethylene films have high slip properties and low density. The thermoplastic film should be thin, less than about 0.1 mm, suitably about 0.010 mm to about 0.050 mm, more suitably about 0.012 mm to about 0.040 mm. Other kinds of films can also be used. Such films include cellulose ethers selected from the group of aliphatic and aromatic ethers; Films in which ethylcellulose is an essential base component, or films of methyl cellulose; Flexible highly plasticized cellulose acetates, formate and similar other alkyl esters; Vinyl vinylidene chloride or rubber hydrochloride such as Pliofilm.TM., Or vinyllite resins.

The thermoplastic film may be transparent or opaque. The film may extend over the entire length of the outer tube, or only extend along a portion thereof. The film may be on the outer layer of the outer tube, or on one of the inner layers.

The layers of the outer tube can be held together by an adhesive, such as a paste, or by heat, pressure, ultrasonic waves or the like. The adhesive can be water soluble or water insoluble. Water soluble adhesives are preferred for environmental reasons that the outer tube will break quickly when impregnated with water. This impregnation will occur when the outer tube is discarded by flushing the toilet.

The outer tube is sized and shaped to accommodate an absorbent menstrual tampon. The inner diameter of the outer tube is sized to accommodate a tampon of typical size. Generally, the inner diameter of the outer tube is less than about 0.75 inches (about 19 mm), more suitably less than about 0.625 inches (about 16 mm). Although the outside diameter of tampons varies, most tampons used by women are less than about 0.75 inches (about 19 mm).

The outer surface of the outer tube should be substantially smooth, which will facilitate insertion of the tampon applicator into the vagina of the woman. If the surface of the outer layer is smooth / soft or smooth, the outer tube will easily slide into the woman's vagina without causing the internal tissue of the woman's vagina to be abrasion. The outer tube can be coated to provide high smoothness to it. Waxes, polyethylenes, blends of waxes and polyethylenes, cellophane and clays are representative coatings that can be applied to the outer layer to facilitate comfortable insertion. The outer tube may be a straight long cylindrical tube formed on the longitudinal axis of the central portion. It is also possible to mold the outer tube into an arcuate shape. The arched or curved shape can assist in providing comfort when inserting the outer tube into the vagina of the woman. With the curved tampon applicator, it is possible to use curved tampons, which may be more comfortable for some women to use, since the shape of the tampon may better fit the curvature of the female vagina.

An insertion tip is integrally formed on the first end of the outer tube and extends out therefrom. The insertion tip is designed to facilitate inserting the outer tube into the vagina of the woman in a comfortable manner. The insertion tip should be made of thin flexible material or membrane, which prevents rapid absorption of vaginal fluid during the period of insertion of the tampon applicator into the vagina of the woman. The insertion tip can be constructed from paper, cardboard or film material. If there are only two layers in the outer tube, the insertion tip should be formed in a layer of lower board weight. Layers with lower board weight are generally thinner layers. Film materials are preferred because they are thin, flexible, and flexible. Suitable materials for the insertion tip include thin bonded nonwoven layers coated with low density polyethylene, plasticized polyvinyl chloride or polyurethane. The insertion tip may also contain a coating or impregnation that inhibits any substantial absorption of the vaginal fluid. The coating can be an oil, a wax, or an acceptable organic compound. Alternatively, the insertion tip can be self-lubricating. Such materials can be made of a polymer that essentially provides an outer surface with a low coefficient of friction. Typical polymers of this type are fluorinated, such as polytetrafluoroethylene (PTFE), fluorinated ethylene-propylene (FEP) and polyethylene oxide (PEO).

The outer diameter of the insertion tip should be approximately equal to or smaller than the outer diameter of the outer tube. If the diameter is smaller than the diameter of the outer tube, it should be noted that the difference should be small so that the female cannot feel the end of the outer layer during insertion. In general, the diameter of the insertion tip is smaller than the diameter of the outer tube. The insertion tip can be shaped into a circular, hemispherical or conical shape. Other nose or dome-shaped shapes may also be used. The circular shape of the insertion tip prevents exhibiting abrasive action in the vaginal wall as in the case where the forward end of the tampon is not covered.

The insertion tip may be formed from one or more of the layers forming the outer tube and may be formed from more than one layer if desired, provided that the thickness is thinner. The insertion tip may be formed from at least one less layer than the number of layers in which the outer tube is built. The thickness of the insertion tip is less than the thickness of the outer tube to ensure that the tip is soft and flexible. The thickness of the insertion tip should be less than about 50% of the thickness of the outer tube, suitably less than about 75% of the thickness of the outer tube, and more suitably less than about 80% of the thickness of the outer tube.

The insertion tip may have a plurality of soft and flexible petals, which are arranged to form a dome-shaped nose. The drawing boards are separated by narrow slots. The drawing board may be radially bent or bent outwards to provide an enlarged outlet through which the tampon can exit when the tampon is pushed forward by the inner tube. Odd or even drawing boards may be used, but odd drawing boards such as 3, 5, 7, etc. exist because odd drawing boards will prevent the outer tube from collapsing or flattening after the tampon is released. By preventing the outer tube from collapse, it can be assured that the vaginal tissue will not be pinched when the tampon applicator is removed from the user's vagina. For example, the insertion tip will each have a long, approximately truncated shape of circular ends, and will contain five drawing boards, each about 7/16 inches (about 11.1 mm) in length.

As mentioned above, the tampon applicator comprises an inner tube. The inner tube, like the outer tube, can be a hollow tube spirally wound, spirally wound or longitudinally sewn from paper, cardboard, cardboard, plastic, film, aqueous coating or combinations thereof. have. The inner tube can also be shaped into a hollow tube by overlaying the material on itself. The inner tube can be constructed from the same material as the outer tube or can be made of a different material. In addition, the inner tube can be constructed as a stack of two or more plies, which are spirally wound, circularly wound or longitudinally sewn into a cylindrical tube. Both wound tubes or longitudinally sewn tubes are preferred because the final tube will have walls of constant thickness. However, some manufacturers may prefer to build the inner tube with a solid stick or use some other unique shape. The inner tube also has a distant or free end on which the user's index finger can be placed to facilitate movement of the inner tube into the outer tube. Thereby the far end acts as a site for the index finger. It is also possible to form an enlarged ring or flange on the far end of the inner tube to provide a larger contact surface.

The inner tube functions by nesting and moving into the outer tube. When the inner tube is pushed into the outer tube, the tampon is pushed forward against the insertion tip. The contact by the tampon causes the drawing board to radially open to a diameter sufficient to allow the tampon to be released from the outer tube. Once tampons are properly placed in the vagina of a woman, the tampon applicator is recovered and discarded.

The weight of the tampon applicator will depend on the size and absorbency of the tampon. For example, longer and better absorbent tampons will be heavier than shorter and less absorbent tampons. Typically, an applicator suitable for use in the present invention with a general absorbent tampon will weigh about 3.62 g; Applicators suitable for use with superabsorbent tampons or superabsorbent tampons will weigh about 4.12 g.

Other suitable non-absorbent articles for the present invention include, for example, non-absorbable incontinence devices, non-absorbable contraceptive devices such as barrier birth control devices, irrigation devices. As used herein, “non-absorbable incontinence device” refers to a device that is designed to be inserted and expanded into the vagina of a woman to mitigate or eliminate the involuntary movement of urine from the bladder across the urethra. Inflation of the non-absorbable urinary incontinence device provides a stable backdrop of muscle and body tissue located around the urethral-vaginal fascia region and allows the urethra to squeeze to itself during episodes of increased internal-abdominal pressure. In addition, expansion of the incontinence instrument in the vagina will assist the urethral sphincter in maintaining a circular cross-sectional shape. One example of a suitable non-absorbable incontinence device is disclosed in US Pat. No. 6,679,831, issued January 20, 2004 to Zunker, et al.

Suitable non-absorbable contraceptive devices such as barrier birth control devices for the present invention are known in the art. For example, US Pat. No. 4,711,235, issued December 8, 1987 to Willis, discloses a mechanism that includes a ring having a central sheet of elastic impermeable material sandwiched between two layers of foam rubber. have. Foam rubber traps sperm and bacteria in the recesses of the vagina for a time sufficient to be destroyed by normal acidic pH of the vaginal secretions. The ring confines the sheet material to a cup shape with four sides, with a pair of opposing sides comprising an inwardly bent wire core, one of the other pair of sides rounded outward and the other fork It has a shape. The elastic impermeable material is typically a neoprene non-absorbent rubber material.

The present invention also relates to absorbent articles and will be described in detail herein with respect to vaginal tampons, but those skilled in the art will appreciate that other disposable absorbent articles such as sanitary napkins, panty liners, adult incontinences that benefit from inhibition of exocrine proteins from Gram-positive bacteria. It will be appreciated that the present invention is applicable to underwear, absorbent contraceptive sponges, diapers, wound dressings, medical bandages and other absorbent tampons such as those intended for medical, dental, surgical and / or nasal applications.

As used herein, the idiom “absorbent tampon” generally refers to vaginal tampons, medical tampons, dental tampons, surgical tampons, and nasal tampons. The phrase “absorbent article” generally refers to an apparatus that absorbs and contains body fluids, and more specifically, absorbs and contains various fluids that are expelled from the body against or near the skin, or within the body cavity. Refers to a mechanism to. The term "disposable" is used herein to describe an absorbent article that is not intended to be washed after use or otherwise restored or reused as an absorbent product. Examples of such disposable absorbent articles include bandages and tampons such as health care-related products, including those intended for medical, dental, surgical and / or nasal use, which are beneficial for inhibiting the production of endocrine proteins from Gram-positive bacteria; Personal hygiene absorbent products such as, but not limited to, feminine hygiene products (eg, sanitary napkins, panty liners and vaginal tampons), diapers, training pants, incontinence products, and the like.

Vaginal tampons suitable for use with the present invention are typically made from absorbent materials, such as absorbent fibers, including natural and synthetic fibers, and compressed into a single body sized for easy insertion into the vaginal steel. Suitable fibers include, for example, cellulose fibers such as cotton and rayon. The fibers can be 100% cotton, 100% rayon, blends of cotton and rayon, or other materials known to be suitable for tampons.

Vaginal tampons are typically made in a long cylindrical form so as to have a sufficiently large body of material that provides the necessary absorbing capacity, but can be made in a variety of shapes. Tampons may or may not be compressed, but the compressed type is generally preferred. Tampons may be made from a variety of fiber blends, including both absorbent and nonabsorbent fibers, which may or may not be encased in a cover or wrap. Typically the cover or sheath can be formed from a nonwoven material such as polyolefin, in particular polypropylene or polyethylene. Suitable materials are spunbond materials. The cover or wrap is advantageous in that it ensures that the fibers of the tampon do not directly contact the female vaginal lining. This ensures that no fibers remain in the vagina after tampons are removed. The cover may be wrapped into the end of the far distance of the tampon so that the fiber is completely enclosed and encased. The cover or wrap may also be constructed from a heat-sealable material that assists in bonding it to the fiber, such as by heat and / or pressure. Suitable methods and materials for the production of tampons are well known to those skilled in the art.

In one embodiment, tampons suitable for use in the present invention have a cover or wrap. Typically, the weight of a tampon with a cover or wrap will depend on the absorbency level of the tampon. For example, more absorbent tampons will be heavier than less absorbent tampons. Typically, the weight of a typical absorbent tampon with a cover or lid will be from about 1.77 g to about 2.67 g, suitably about 2.22 g; The superabsorbent tampon with the cover or lid will weigh about 2.67 g to about 3.57 g, suitably about 3.12 g; The superabsorbent tampon with the cover or lid will weigh about 3.67 g to about 4.97 g, suitably about 4.32 g.

Tampons vary in size. In another embodiment, tampons for use in the present invention do not have a cover or wrap. Typically, a typical absorbent tampon without a cover or wrapper suitable for the present invention will weigh from about 1.60 g to about 2.50 g, suitably about 2.05 g; The superabsorbent tampon without cover or wrap will weigh from about 2.49 g to about 3.39 g, suitably about 2.94 g; The superabsorbent tampon without cover or wrap will weigh from about 3.49 g to about 4.79 g, suitably about 4.14 g.

As mentioned above, the non-absorbable and absorbent articles of the present invention, when hydrolyzed, have the activity capable of substantially inhibiting the production of exocrine proteins by Gram-positive bacteria, in particular the production of TSST-1 from Staphylococcus aureus bacteria. An effective amount of precursor compound to produce the species. As used herein, the term “precursor compound” is used in non-absorbable or absorbent articles and can be hydrolyzed within and / or around the vagina to produce active species that can inhibit exocrine protein production from Gram-positive bacteria. And / or a compound introduced onto such an article. The precursor compound is formed by linking the aromatic compound to the second compound via an ester or amide bond. Ester or amide bonds contained in the precursor compound are hydrolyzed by enzymes produced by bacteria found in the natural vaginal flora, such as lipases, esterases and / or amidases, to produce exocrine protein production from Gram-positive bacteria. Active species that can be inhibited can be produced. With the active species produced, hydrolysis reactions produce a second compound which is not critical to the function of the active species. In some embodiments, the second compound will be the same or similar to the compound naturally occurring in the human body. As mentioned above, although the second compound is not critical, the precursor compound should be designed so that the second compound formed upon hydrolysis is substantially harmless to the vagina or bacteria located in the vagina.

During the hydrolysis process, the precursor compound slowly degrades into the active species and the second compound; As mentioned above, both the precursor compound and the active species can inhibit the production of exocrine proteins from Gram-positive bacteria. This property is advantageous because it allows long-term, continuous inhibition of exocrine protein production by Gram-positive bacteria. The precursor compounds of the present invention can inhibit the production of exocrine proteins prior to hydrolysis; Thereafter, when the precursor compound is hydrolyzed, the active species can be produced, which can further inhibit the production of the exocrine protein.

Precursor compounds described herein and suitable for introduction onto and / or into a non-absorbent or absorbent article are both within and / or within the non-absorbent or absorbent article, throughout the manufacturing process and during storage. Both phases are substantially stable. In other words, the precursor compound does not readily volatilize away from or within the non-absorbent or absorbent article during the hot manufacturing process or during transportation and storage. This property of the precursor compound is highly desirable because it is important that the effective amount of the precursor compound remain in or on the non-absorbent or absorbent article product for end use by the consumer. As mentioned above, in the past, the volatility of the active compounds from non-absorbent or absorbent articles has been a problem, resulting in non-absorbent articles or absorbent articles free of active compounds.

Without being limited to a particular theory, it is believed that the precursor compounds of the present invention may remain in or on non-absorbent or absorbent articles in increased amounts relative to existing components due to the increased molecular weight. Although the molecular weight of the precursor compounds of the invention is generally higher compared to some of the active ingredients used in the past, the precursor compounds of the invention are adequately hydrolyzed in the body to produce highly preferred active species such as benzyl alcohol and benzoic acid. They are highly effective at inhibiting the production of exocrine proteins by Gram-positive bacteria.

In addition to reducing volatility and being able to remain in and / or on non-absorbing or absorbent articles throughout manufacture, transportation, and storage, the precursor compounds described herein are hydrolyzed active species produced in the body. And, along with the second compound, does not kill a significant amount of naturally occurring bacteria found in the vagina. This property is important because a natural bacterial flora is necessary to maintain a healthy vagina, because completely or substantially completely non-specific killing of bacteria located in and around the vagina can be very harmful to the host. In contrast to antimicrobial or antiviral agents that non-specifically kill all microorganisms in the vagina, the precursor compounds and hydrolyzed compounds produced in and around the vaginal cavity have a substantial killing effect on bacteria at concentrations incorporated into the product. Do not have; Active species produced by hydrolysis can substantially inhibit the production of exocrine proteins from Gram-positive bacteria.

In one embodiment of the invention, the precursor compound has the formula:

이고; R⁷은 -OCH₂-이고; X는 0 또는 1이고; R⁵는 치환 또는 비치환 방향족 고리, 또는 헤테로 원자로 치환되거나 치환되지 않을 수 있는, 1가의 포화 또는 불포화, 치환 또는 비치환, 분지쇄 또는 직쇄 히드로카르빌 잔기이고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is

ego; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

As mentioned above, the hydrocarbyl residues described herein include both straight and branched chain hydrocarbyl residues which may or may not be substituted with various substituents such as, for example, hydroxyl groups. In addition, the hydrocarbyl residue may or may not involve a hetero atom. Heteroatoms that may be involved in the hydrocarbyl residue include, for example, oxygen, nitrogen and sulfur. In one embodiment, R ⁵ is 1 to 15 carbon atoms, more suitably 1 to 12 carbon atoms. Monovalent saturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue with

Some specific precursor compounds having ester bonds and suitable for the above described embodiments of the invention can be found in Table 1 below.

compound Chemical formula Benzyl (s)-(-) lactate

Benzyl ethyl malonate

Benzyl laurate

Benzyl benzoate

Benzyl paraben

Benzyl salicylate

Phenoxy-ethyl paraben

The compounds may be used alone or in combination.

As mentioned herein, enzymes such as esterases produced by bacteria such as Staphylococcus aureus found in the vaginal flora, along with enzymes that occur naturally in menstrual fluid, hydrolyze the precursor compounds described herein to produce active species. And a second compound. For example, esterase enzymes can be reacted with ester bonds of the precursor compounds described above to form active species benzyl alcohol and hydrocarbons. Benzyl alcohol has been found to substantially inhibit exocrine proteins from Gram-positive bacteria without substantially removing Gram-positive bacteria.

In another embodiment, the precursor compound has the formula:

이고; R⁶은 아미노산, 아미노산의 메틸 에스테르, 및 아미노산의 에틸 에스테르로 구성되는 군으로부터 선택되고; R², R³ 및 R⁴는 H, OH, COOH로 구성되는 군으로부터 독립적으로 선택된다].[Wherein R ¹ is

ego; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH.

An amino acid is an organic compound containing an amino group and a carboxylic acid group. Suitable amino acids that can be used for R ⁶ are all 20 amino acids found naturally in the human body. More particularly, amino acids suitable for use in the present invention include, for example, valine, leucine, cysteine and combinations thereof.

Suitable compounds for this embodiment can be found in Table 2 below.

compound Chemical formula N-benzoyl-DL-valine

N-benzoyl-DL-leucine

N-benzoyl-DL-cysteine

The compounds described herein can be used alone or in combination.

As described above, enzymes such as amidase produced by bacteria such as Staphylococcus aureus found in the vaginal flora can hydrolyze precursor compounds containing amino acids with enzymes that occur naturally in menstrual fluid. For example, amidase enzymes can react with precursor compounds to cleave the amide bonds of the compounds of this embodiment, releasing benzoic acid and amino acids. Like benzyl alcohol discussed above, benzoic acid has been found to substantially inhibit exocrine proteins from Gram-positive bacteria without substantially removing naturally occurring bacterial flora.

In accordance with the present invention, non-absorbable or absorbent articles, when hydrolyzing a precursor compound, produce a sufficient amount of active agent such that an endocrine protein such as TSST-1 is present when the non-absorbable or absorbent article is exposed to Staphylococcus aureus bacteria. An effective amount of precursor compound is included so that the formation of is substantially inhibited. Various methods are known in the art for testing the effectiveness of potential inhibitors, such as benzyl alcohol or benzoic acid, for inhibiting TSST-1 production in the presence of Staphylococcus aureus. One such preferred method is described in Example 1. When tested according to the methods described herein, preferably, the active species produced from the hydrolysis of the precursor compound exhibits at least about 40%, more preferably at least about 50%, formation of TSST-1 by Staphylococcus aureus, Even more preferably at least about 60%, even more preferably at least about 70%, even more preferably at least about 80%, even more preferably at least about 90%, even more preferably at least about 95% reduction Let's do it. Additionally, as mentioned above, precursor compounds may also inhibit the production of exocrine proteins from Gram-positive bacteria in some embodiments. The test procedure described in Example 1 can also be used to determine the amount of inhibition by the precursor compound. In some embodiments, the precursor compound may reduce at least about 50%, preferably at least about 80%, TSST-1 formation.

In accordance with the present invention, non-absorbable or absorbent articles, when used, substantially inhibit the production of exocrine proteins from Gram-positive bacteria with precursor compounds and / or active species produced in or around the vagina as discussed herein. It may contain an appropriate amount of precursor compound to be inhibited. Generally, the non-absorbent product will comprise from about 0.15% (by weight of the non-absorbent substrate) to about 2.0% (by weight of the non-absorbent substrate) and the absorbent article will contain about 0.15% (absorbent structure). By weight); and from about 2.0% (by weight of absorbent structure). Such percentages are commonly referred to as "add on weight percentages". In a preferred embodiment, the non-absorbent product will comprise from about 0.17% (by weight of the non-absorbent substrate) to about 1.7% (by weight of the non-absorbent substrate) and the absorbent product will contain about 0.17% ( By weight of absorbent structure) to about 1.7% (by weight of absorbent structure).

In one particular embodiment, a tampon applicator as described herein, when used, comprises a precursor compound as discussed herein, and / or an active species produced in or around the vagina from an exocrine protein from Gram-positive bacteria. An appropriate amount of precursor compound is included to substantially inhibit production. In general, the tampon applicator will comprise from about 0.15% (by weight of the non-absorbing substrate) to about 2.0% (by weight of the non-absorbing substrate). In a preferred embodiment, the tampon applicator will comprise from about 0.17% (by weight of the non-absorbing substrate) to about 1.7% (by weight of the non-absorbing substrate).

In another particular embodiment, vaginal tampons as described herein, without a cover or wrap material, are, when in use, a precursor compound as discussed herein and / or an active species produced in or around the vagina from the precursor compound as discussed herein. An appropriate amount of precursor compound is included so as to substantially inhibit the production of exocrine proteins from positive bacteria. Generally, vaginal tampons without cover or wrap material will comprise from about 0.15% (by weight of absorbent tampon material) to about 2.0% (by weight of absorbent tampon material). In a preferred embodiment, the vaginal tampon will comprise from about 0.17% (by weight of the absorbent tampon material) to about 1.7% (by weight of the absorbent tampon material).

In another embodiment, in an amount suitable for use, the precursor compound as discussed herein and / or active species produced therein or around the vagina can substantially inhibit the production of exocrine proteins from Gram-positive bacteria. A vaginal tampon as described herein is provided having a cover or sheath material, comprising a precursor compound of. The precursor compound is introduced into the cover or wrapper or directly onto the cover or wrapper as opposed to being introduced into or on the absorbent substrate of the vaginal tampon. In a preferred embodiment, the cover or wrap material of the vaginal tampon will comprise from about 2.6% (by weight of the cover or wrap material) to about 35.0% (by weight of the cover or wrap material). More preferably, the cover or wrap material of the vaginal tampon will comprise from about 2.95% (by weight of the cover or wrap material) to about 29.5% (by weight of the cover or wrap material).

In a preferred embodiment of the invention, the precursor compounds described herein are non-absorbent articles in combination with one or more surfactants to further reduce the production of endocrine proteins such as TSST-1 without substantially removing the beneficial bacterial flora. Or into an absorbent article and / or on a non-absorbent article or absorbent article. Surfactants used in combination with precursor compounds may also act as lubricants and / or softeners to further improve product performance. When used in combination with vaginal tampons, surfactants may further assist in the removal of "dry tampons". In one embodiment where a tampon applicator or vaginal tampon is included, a suitable surfactant is mireth-3-myristate, which is a Kraft Chemical Corp. (Melrose Park, Illinois) available as CETIOL 1414. Other surfactants suitable for the present invention include, for example, glycerol monolaurate and laureth-4.

According to the present invention, a non-absorbing or absorbing article may comprise a sufficient amount of surfactant such that, in use, the surfactant may further inhibit the production of exocrine proteins from Gram-positive bacteria. Generally, the non-absorbent product will include from about 0.4% (by weight of the non-absorbent substrate) to about 1.1% (by weight of the non-absorbent substrate). In one particular embodiment, the non-absorbent product is a tampon applicator comprising about 0.75% (by weight of the non-absorbent substrate) surfactant. Generally, the absorbent article will include from about 0.4% (by weight of the absorbent structure) to about 1.1% (by weight of the absorbent structure). In one particular embodiment, the absorbent article is a vaginal tampon with a cover and will comprise about 0.75% (based on the weight of the total tampon including the cover) of the surfactant.

In another embodiment, the surfactant component may be introduced into the cover or wrap material or directly onto the cover or wrap material as opposed to being introduced into or on the absorbent substrate of the vaginal tampon. Typically, the cover or wrap material of the vaginal tampon will comprise about 13% (based on the weight of the cover material) of the surfactant.

Additionally, the precursor compounds described herein may be combined with one or more second agents to further reduce the production of endocrine proteins such as TSST-1 without substantially removing the beneficial bacterial flora and / or into a non-absorbing or absorbing product. Or on a non-absorbent or absorbent article. Suitable examples of second agents useful in the present invention include agents selected from the group consisting of ethers, esters, amides, glycosides, or compounds having amine bonds that link C ₈ -C ₁₈ fatty acids to aliphatic alcohols.

In one embodiment, the precursor compounds described herein can be used in combination with ester compounds of the formula:

Wherein R ²⁷ is linear or branched alkyl or linear or branched alkenyl having 8 to about 18 carbon atoms and R ²⁸ is selected from the group consisting of alcohols, polyhydric alcohols and ethoxylated alcohols. The term "multivalent" as used herein refers to the presence of two or more hydroxyl (OH) groups in a compound. Suitable compounds include glyceryl monolaurate, glyceryl dilaurate, mireth-3-myristate, and mixtures thereof.

In another embodiment, the precursor compounds described herein can be used in combination with ether compounds of the formula:

[Wherein R ¹⁰ is linear or branched alkyl or linear or branched alkenyl having 8 to about 18 carbon atoms, R ¹¹ is alcohol, ethoxylated alcohol, polyalkoxylated sulfate salt and polyalkoxylated sulfosuccisine Nate salts. Suitable compounds include laureth-3, laureth-4, laureth-5, PPG-5 lauryl ether, 1-0-dodecyl-rac-glycerol, sodium laureth sulfate, potassium laureth sulfate, di Sodium laureth (3) sulfosuccinate, dipotassium laureth (3) sulfosuccinate, and polyethylene oxide (2) sorbitan ether.

In another embodiment, the precursor compounds described herein can be used in combination with alkyl polyglycoside compounds. Suitable alkyl polyglycosides for use in combination with precursor compounds include alkyl polyglycosides of the formula:

Wherein Z is a saccharide having 5 or 6 carbon atoms, n is an integer from 1 to 6, and R ¹⁴ is a linear or branched alkyl group having from about 8 to about 18 carbon atoms. Glucopons 220, 225, 425, 600, and 625 (all commercially available from Henkel Corporation (Ambler, Pennsylvania)) and TL 2141 (commercially available from ICI Surfactants (Wilmington, Delaware)) Alkyl polyglycosides suitable for use in combination.

In another embodiment, the precursor compounds described herein can be used in combination with amide containing compounds of the formula:

[Wherein R ¹⁷ including carbonyl carbon is an alkyl group having 8 to 18 carbon atoms, R ¹⁸ and R ¹⁹ are independently hydrogen or an ester group, an ether group, an amine group, a hydroxyl group, a carboxyl group, Selected from carboxyl salts, sulfonate groups, sulfonate salts, and mixtures thereof, and alkyl groups having 1 to about 12 carbon atoms which may or may not be substituted. Preferred amide compounds for use in combination with the precursor compounds described herein include sodium lauryl sarcosinate, lauryl monoethanolamide, lauryl diethanolamide, lauamidopropyl dimethylamine, disodium lauric Amido monoethanolamide sulfosuccinate, and disodium lauroampodiacetate.

In another embodiment, the precursor compounds described herein can be used in combination with amine compounds of the formula:

[Wherein R ²⁰ is an alkyl group having from about 8 to about 18 carbon atoms, R ²¹ and R ²² independently have hydrogen and 1 to about 18 carbon atoms, and hydroxyl, carboxyl, carboxyl salts, And an alkyl group which may have one or more substitution moieties selected from the group consisting of imidazolines. Preferred amine compounds for use in combination with the precursor compounds described herein include triethanolamide laureth sulfate, laumine, lauamino propionic acid, sodium lauriminodinodipropionic acid, lauryl hydroxyethyl imidazoline and these Mixtures thereof.

In another embodiment, the amine compound can be an amine salt of the formula:

[Wherein R ²³ is an anionic moiety associated with an amine and is derived from an alkyl group having from 8 to about 18 carbon atoms, R ²⁴ , R ²⁵ , and R ²⁶ represent hydrogen, and from 1 to about 18 carbon atoms Independently selected from the group consisting of an alkyl group which may have at least one substituent residue selected from the group consisting of hydroxyl, carboxyl, carboxyl salt, and imidazoline. R ²⁴ , R ²⁵ , and R ²⁶ may be saturated or unsaturated. Exemplary preferred compounds of the amine salts are TEA laureth sulfate.

The amount of second compound described herein added to the non-absorbent product to further reduce the production of TSST-1 ranges from about 0.15% (by weight of the non-absorbing substrate) to about 2.0% (by weight of the non-absorbing substrate) It was found to be the second compound of). More suitably, the non-absorbent product comprises from about 0.17% (by weight of the non-absorbing substrate) to about 1.7% (by weight of the non-absorbing substrate).

The amount of the second compound described herein added to the absorbent article to further reduce the production of TSST-1 is from about 0.15% (by weight of the absorbent structure) to about 2.0% (by weight of the absorbent structure) Was found. More suitably, the absorbent article includes from about 0.17% (by weight of the absorbent structure) to about 1.7% (by weight of the absorbent structure).

The precursor compound can be applied to the non-absorbing substrate using conventional methods for applying the chemical agent to the desired non-absorbing substrate. For example, the non-absorbent article can be directly immersed into a liquid bath containing the precursor compound and then, if necessary, air dried to remove any volatile solvent. Alternatively, non-absorbent articles of the invention can be sprayed or otherwise coated with the inhibitory compounds of the invention.

One or more pharmaceutically acceptable and compatible carrier materials useful for the desired application, such as to facilitate absorption into absorbent articles, may additionally be used in the precursor compounds. Precursor compounds used in non-absorbent or absorbent articles may be co-dissolved or suspended in the carrier. Carrier materials suitable for use in the present invention include those well known for use in the cosmetic and medical industry as base materials for ointments, lotions, creams, plasters, aerosols, suppositories, gels and the like. For example, one suitable carrier is cetiol. In addition, as discussed above, cethiol may also function as both a lubricant and a softener. Other suitable carrier materials include water, various alcohols, and other organic solvents.

In another embodiment, the precursor compounds of the invention can be formulated into a variety of formulations such as those currently available in irrigation formulations or higher viscosity irrigation apparatuses. For example, the precursor compounds of the present invention can be incorporated into formulations used to irrigate the vagina to prevent vaginal infections. In addition, the precursor compound may be combined with a surfactant, preferably a nonionic surfactant such as Cremophos RH60, Tween 20 and the like. Formulations of the present invention may also contain a preservative. Compounds that can give better viscosity, such as propylene glycol, may be added to the formulations of the present invention. In general, higher viscosity formulations are preferred to produce formulations that tend to remain in the vagina for a relatively long time after administration.

When the precursor compound is incorporated into a formulation comprising a pharmaceutically acceptable carrier, the formulation is typically at least about 0.01% (weight / volume), preferably at least about 0.4% (weight / volume) of the precursor compound (the entirety of the formulation By weight). Generally, formulations contain up to about 0.3% (weight / volume) of precursor compounds. Particularly suitable formulations for use in vaginal cleaning applications include at least about 0.2 mM / L to about 50 mM / L, suitably about 0.3 mM / L to about 30 mM / L, more suitably about 1.0 mM / L to about May contain 15 mM / L of precursor compound.

Precursor compounds of the invention may be prepared and applied in any suitable form of absorbent article, but preferably include, but are not limited to, aqueous solutions, lotions, balms, gels, plasters, ointments, bolus, suppositories, and the like. It is manufactured in the form.

The precursor compound can be applied to the absorbent article using conventional methods for applying a chemical agent to the desired absorbent article. For example, a single tampon without a separate wrap may be sprayed with the desired concentration of precursor compound and then air dried if necessary to remove any volatile solvent. For compressed tampons, it is generally desirable to impregnate tampons with any compound prior to compression. Precursor compounds when incorporated onto and / or into tampon materials may be fugitive, loosely attached, bound, or a combination thereof. The term "temporary" as used herein, means that the composition can move through tampon material.

Generally, it is not necessary to impregnate the precursor compound into the overall absorbent of tampons. Economically and functionally optimal results can be obtained by concentrating the precursor compound on or around the exterior surface of tampons or other absorbent articles that may be most effective during use.

In some embodiments, additional ingredients commonly found in pharmaceutical compositions can be used in combination with the precursor compounds of the invention in a manner established in the art and at concentrations that will not alter the normal vaginal flora. For example, the composition may contain additional compatible pharmaceutically active substances for combination therapy, such as adjuvant selective antimicrobials, antioxidants, repellents, anti-itches, astringents, local anesthetics, or anti-inflammatory agents.

In one embodiment, the precursor compound may be microencapsulated in a shell-like material that breaks down, disintegrates, ruptures, or otherwise breaks upon contact with menses or other vaginal secretions to secrete components. In such embodiments, the encapsulating material is wetted with humoral secretions to delay volatilization of the precursor compound until the release of the precursor compound is caused. Such encapsulation significantly increases the amount of precursor compound present in the product after fabrication and storage. Suitable microencapsulated shell materials are known in the art and include cellulosic polymeric materials (eg ethyl cellulose), carbohydrate-based materials (eg cationic starch and sugars) and materials derived from them (eg Dextrins and cyclodextrins) as well as other materials compatible with human tissues.

The microencapsulated shell thickness can vary and is generally prepared so that the encapsulated precursor compound is covered with a thin layer of encapsulation material, which layer can be a single layer or a thicker stack, or can be a composite layer. The microencapsulation layer should be thick enough to withstand cracking or breakage of the shell during handling or transportation of the product. The microencapsulation layer should be constructed so that humidity from atmospheric conditions during storage, transportation, or wear does not cause the microencapsulation layer to break and release the precursor compound.

Microencapsulated formulations or ingredients applied directly to non-absorbable or absorbent articles should be sized such that the user cannot feel the encapsulated shell on the skin or mucous membranes during use. Typically, the diameter of the capsule is about 25 μm or less, preferably about 10 μm or less. At this size, there is no feeling of "gritty" or "scratchy" when the formulation is in contact with the skin.

The invention is illustrated by the following examples, which are for illustrative purposes only and are not to be construed as limiting the scope of the invention or the manner in which the invention may be practiced.

Example  One

In this example, the effect of various test compounds on the growth of Staphylococcus aureus and TSST-1 production was determined. Test compounds at the desired concentration (expressed in weight percent (weight / volume)) were placed in 10 ml growth medium in conical sterile 50 ml polypropylene tubes (Sarstedt, Inc., Newton, North Carolina).

Growth medium was prepared by dissolving 37 g of brain heart infusion broth (BHI) (Difco Laboratories (Cockeysville, Maryland)) in 880 ml of distilled water and sterilizing the broth according to the manufacturer's instructions. BHI was supplemented with 100 ml fetal bovine serum (FBS) (Sigma Chemical Company (St. Louis, Missouri)). 10 ml of a 0.021M sterile solution of magnesium chloride hexahydrate (Sigma Chemical Company (St. Louis, Missouri)) was added to the BHI-FBS mixture. 10 mL of 0.027M sterile solution of L-glutamine (Sigma Chemical Company (St. Louis, Missouri)) was also added to the BHI-FBS mixture.

Compounds to be tested included N-benzoyl-DL-leucine (Sigma B-1504) and N-benzoyl-DL-valine (Sigma B-6500). Test compound was provided as a solid. The solid was dissolved in BHI prepared as described above. Test compounds were added to the growth medium in the amount necessary to obtain the desired final concentration. Cetiol 1414E (Mireth-3-myristate) (Kraft Chemical Corp. (Melrose Park, Illinois)) was included in the growth medium at a concentration of 10 mM in some assays.

In preparation for inoculation of tubes of growth medium containing test compounds, inoculation broths were prepared as follows: Staphylococcus MN8 was tryptic soy agar plate (TSA; Difco Laboratories (Cockeysville, Maryland)) Streaked onto phase and incubated at 35 ° C. Test organisms in this example were obtained from Dr. Pat Schlievert (Department of Microbiology, University of Minnesota Medical School, Minneapolis, Minnesota). After 24 hours of incubation, 3 to 5 individual colonies were picked up with a sterile inoculation loop and inoculated in 10 ml of growth medium. Tubes of inoculated growth medium were incubated at 35 ° C. in atmospheric air. After 24 hours of incubation, the cultures were removed from the incubator and mixed well on an S / P brand vortex mixer. A second tube containing 10 ml of growth medium was inoculated with 0.5 ml of the above described 24-hour culture and incubated at 35 ° C. in atmospheric air. After 24 hours of incubation, the cultures were removed from the incubator and mixed well on an S / P brand vortex mixer. The optical density of the culture was determined in a microplate reader (Bio-Tek Instruments (Winooski, Vermont), Model EL309). The amount of inoculum required to provide 5 × 10 ⁶ CFU / ml in 10 ml growth medium was determined using standard curves prepared previously.

This example included a tube of growth medium containing varying concentrations of test compound, a tube of growth medium containing varying concentrations of test compound and cethiol 1414E, and a tube of growth medium without test compound (control). Each tube was inoculated with a determined amount of inoculum as described above. The tube was closed with a foam plug (IDENTI-PLUG plastic foam plug from Jaece Industries, purchased from VWR Scientific Products (South Plainfield, New Jersey)). The tube was incubated at 35 ° C. in atmospheric air containing 5% by volume CO ₂ . After 24 hours of incubation, the tubes were removed from the incubator and the cultures analyzed for the number of colony forming units of Staphylococcus aureus and cultures prepared for TSST-1 analysis as described below.

The number of colony forming units per ml after incubation was determined using standard plate counting procedures. In preparation for TSST-1 analysis, the culture broth was centrifuged at 2500 rpm for 15 minutes at 2-10 ° C., and the supernatant was then filter sterilized through a 0.2 μm pore sized FISHERBRAND 0.45 μm MCE filter. The resulting fluid was frozen at −70 ° C. in a FISHERBRAND 12 × 75 mm polystyrene culture tube (Fisher Scientific (Pittsburgh, Pennsylvania)).

The amount of TSST-1 per ml was determined by non-competitive sandwich enzyme linked immunosorbent assay (ELISA). The method used was as follows: TSST-1 (# TT-606), rabbit polyclonal anti-TSST-1 IgG (LTI-101), rabbit polyclonal anti-TSST conjugated to horseradish peroxidase Four reagents of -1 IgG (# LTC-101), and normal rabbit serum (NRS) (# NRS-10) that were found to be free of anti-TSST-1 were selected from Toxin Technology, Inc. (Sarasota, Florida). A 10 μg / ml solution of polyclonal rabbit anti-TSST-1 IgG was prepared in phosphate buffered saline (PBS), pH 7.4. PBS was prepared from 0.016 M NaH ₂ PO ₄ , 0.004 M NaH ₂ PO ₄ -H ₂ O, 0.003 M KCl and 0.137 M NaCl (all available from Sigma Chemical Company (St. Louis, Missouri)). 100 μl of polyclonal rabbit anti-TSST-1 IgG solution was pipetted into the inner wells of polystyrene microplates (Nunc-Denmark, Cat # 439454). The plate was covered and incubated overnight at room temperature. Drain until dry to remove unbound anti-toxin.

Phosphate buffered saline (PBS) containing 0.05% (volume / volume) Tween-20 (pH 7.4) (PBS-Twin) (Sigma Chemical Company (St. Louis, Missouri)) and 1% (Volume / Volume) Dilute to 10 ng / ml with NRS and incubate overnight at 4 ° C. Test samples were combined with 1% NRS (volume / volume) and incubated overnight at 4 ° C. Samples of the culture and TSST-1 reference standards were analyzed in triplicate.

100 μl of a 1% (weight / volume) solution of the sodium salt of casein (Sigma Chemical Company (St. Louis, Missouri)) in PBS was pipetted into the inner wells of a polystyrene microplate. The plate was covered and incubated at 35 ° C. for 1 hour. Wash three times with PBS-Twin to remove unbound BSA. TSST-1 reference standards (10 ng / ml) treated with NRS, test samples treated with NRS, and reagent controls were pipetted into respective wells on rows 1 and 7 of the plates in 200 μl volume. 100 μl of PBS-Tween was added to the remaining wells. Thereafter, 100 μl of TSST-1 reference standard and test sample were transferred from the wells to the wells and serially diluted five times in PBS-twin. Samples were mixed prior to migration by repeated aspiration and expression. Test samples and samples of TSST-1 reference standards were analyzed in triplicate. It was then incubated at 35 ° C. for 1.5 hours and washed five times with PBS-T and three times with distilled water to remove unbound toxins. Rabbit polyclonal anti-TSST-1 IgG conjugated to horseradish peroxidase was diluted according to the manufacturer's instructions and 50 μl was added to each microtiter well except well A-1, the conjugate control well. The plate was covered and incubated at 35 ° C. for 1 hour.

After incubation, the plates were washed 5 times with PBS-Twin and 3 times with distilled water. After washing, the wells were horseradish consisting of 5 μl 30% hydrogen peroxide and 5 mg o-phenylenediamine in 11 mL citrate buffer (pH 5.5), both available from Sigma Chemical Company (St. Louis, Missouri). Treatment with 100 μl peroxidase substrate buffer. Citrate buffer was prepared from 0.012 M citric anhydride and 0.026 M dibasic sodium phosphate, both available from Sigma Chemical Company (St. Louis, Missouri). Plates were incubated at 35 ° C. for 15 minutes. The reaction was stopped by adding 50 μl of 5% sulfuric acid solution. The intensity of the color reaction in each well was evaluated using a Bio-Tek model EL309 microplate reader (OD 490 nm). TSST-1 concentrations in the test samples were determined from the reference toxin regression equation derived during each assay procedure. The efficacy of the compounds in inhibiting TSST-1 production is shown in Table 3 below.

Amino acid-containing compounds Amount of Test Compound (wt%) Cetiol 1414E CFU / mL ELISA: TSST-1 ng / ml (% inhibition) none none 2.63E + 0.8 178.4 (N / A) N-benzoyl-DL-leucine 0.25% (weight / volume) none 2.69E + 0.8 14.6 (92.8%) N-benzoyl-DL-valine 0.20% (weight / volume) none 1.61E + 0.8 25.1 (79%) none 10 mM 1.75E + 0.8 53.1 (60%) N-benzoyl-DL-leucine 0.25% (weight / volume) 10 mM 2.23E + 0.8 7.9 (95%) N-benzoyl-DL-valine 0.20% (weight / volume) 10 mM 2.07E + 0.8 10.6 (93.2%) N / A = not applicable

In accordance with the present invention, the data in Table 3 indicate that Staphylococcus MN8 produced less TSST-1 in the presence of test compounds containing amino acids compared to the control. At the concentrations tested, these compounds reduced the amount of toxin produced by 79% to 93%. The data also show that Staphylococcus MN8 produced less TSST-1 in the presence of test compounds containing amino acids when combined with Cetiol 1414E compared to the control. At the concentrations tested, these compounds reduced the amount of toxin produced when combined with cethiol 1414E, between 93% and 95%. However, although the amount of toxin produced was significantly reduced, the effect on the growth of Staphylococcus aureus was minimal, if any.

Example  2

In this example, the effects of benzyl alcohol (Aldrich 40,283-4) and benzyl ethyl malonate (Aldrich 30,069-1) (Sigma Chemical Corporation (St. Louis, Missouri)) on the growth of Staphylococcus aureus and TSST-1 production Was determined. The effect of the test compound tested in Example 2 was determined by placing the desired concentration, expressed in% (volume / volume), in 10 ml of growth medium as described in Example 1. Thereafter, the test compound was tested and evaluated as in Example 1 except that each test was performed in triplicate. The results presented represent the average of four values. The effect of test compounds on the growth and Staphylococcus aureus MN8 production and TSST-1 production is shown in Table 4 below.

compound Amount of test compound (% (volume / volume)) Cetiol 1414E CFU / mL ELISA: TSST-1 ng / ml (% inhibition) none none 7.36E + 0.8 338 (N / A) Benzyl alcohol 0.3% none 7.52E + 0.8 50 (85%) Benzyl ethyl malonate 0.8% none 3.78E + 0.8 19 (94%) none 10 mM 2.61E + 0.8 70 (79%) Benzyl alcohol 0.3% 10 mM 5.02E + 0.8 4 (99%) Benzyl ethyl malonate 0.8% 10 mM 1.51E + 0.8 6 (98%) N / A = not applicable

At the concentrations tested, both compounds of benzyl ethyl malonate and the active species benzyl alcohol reduced the amount of toxin produced by 85% to 94%. The data also show that Staphylococcus MN8 produced less TSST-1 in the presence of benzyl alcohol or benzyl ethyl malonate when combined with cethiol 1414E (mireth-3-myristate) compared to the control. . At the concentrations tested, these compounds, when combined with cethiol 1414E, reduced the amount of toxin produced 98-99%.

Statistical analysis of treatments was performed using pairwise comparisons to Least Squares Means in the Analysis of Variance environment. Pair comparison is equivalent to the standard T test. Comparison results show that growth with Staphylococcus alone is the growth of Staphylococcus aureus in the absence of cethiol or in the presence of only benzyl alcohol or with benzyl ethyl malonate in the presence or absence of cethiol. Significantly more suppressed. Growth in the presence of benzyl ethyl malonate and growth in the presence of a combination of cethiol and benzyl alcohol or benzyl ethyl malonate significantly reduced toxin production compared to growth in the presence of cetiol alone or growth in the absence of additives. . However, although the amount of toxin produced was significantly reduced under these conditions, the effect on the growth of Staphylococcus aureus was minimal, if any.

Example  3

In this example, the effect of benzyl (s)-(-)-lactate (Aldrich 42,484-6) (Sigma Chemical Corporation (St. Louis, Missouri)) on the growth of Staphylococcus aureus and TSST-1 production was determined. It was. The effect of the test compound was determined by placing the desired concentration, expressed in% (volume / volume), in 10 ml of growth medium as described in Example 1. Thereafter, the test compound was tested and evaluated as in Example 1 except that each test was performed in triplicate. The results presented represent the average of four values. The effect of test compounds on the growth of Staphylococcus aureus MN8 and TSST-1 production is shown in Table 5 below.

compound Amount of test compound (% (volume / volume)) Cetiol 1414E CFU / mL ELISA: TSST-1 ng / ml (% inhibition) none none 4.60E + 0.8 604 (N / A) Benzyl (s)-(-)-lactate 0.3% none 4.86E + 0.8 18 (96.9%) none 10 mM 2.48E + 0.8 61 (89.9%) Benzyl (s)-(-)-lactate 0.3% 10 mM 1.48E + 0.8 5 (99.2%) N / A = not applicable

At the tested concentrations, benzyl (s)-(-)-lactate reduced the amount of toxin produced by 97%. In addition, the data indicate that Staphylococcus MN8 was less TSST-1 in the presence of benzyl (s)-(-)-lactate when combined with cethiol 1414E (mireth-3-myristate) compared to the control. Indicates production. At the tested concentrations, benzyl (s)-(-)-lactate, when combined with cethiol 1414E, reduced the amount of toxin produced by 99%.

Statistical comparisons of treatments were performed by the method discussed in Example 2. Comparison results show that growth in Staphylococcus aureus growth in the absence of cetiol or growth in the presence of cethiol or growth in the presence of cethiol and benzyl (s)-(-)-lactate or benzyl (s)-(- It was shown to be significantly more inhibited than growth with) -lactate alone. Growth in the absence of additives or in the absence of additives, growth in the presence of benzyl (s)-(-)-lactate and growth in the presence of a combination of cethiol and benzyl (s)-(-)-lactate Compared to growth, toxin production was significantly reduced.

Example  4

In this example, the effect of benzyl (s)-(-)-lactate in combination with surfactant Cetiol 1414E (mireth-3-myristate) was tested using a 5 × 4 checkerboard experimental design. This evaluated the interaction of the two test compounds with Staphylococcus aureus growth and TSST-1 production.

Sorting 20 tubes, 5 concentrations of benzyl (s)-(-)-lactate (0.00%, 0.06%, 0.13%, 0.25%, and 0.50%) with 4 concentrations of Cetiol 1414E (10 mM, 5 mM, 2.5 mM, and 0.0 mM). For example, Tube # 1 contained 0.0 mM cethiol 1414E and 0.0% benzyl (s)-(-)-lactate (weight / volume) in 10 ml of growth medium (as prepared in Example 1). . Each tube # 1- # 20 contained a unique combination of benzyl (s)-(-)-lactate and cethiol. The solution was tested and evaluated as in Example 1. The effect of test compounds on the growth of Staphylococcus aureus MN8 and TSST-1 production is shown in Table 6 below.

Cetiol mM Benzyl (s)-(-)-lactate% (weight / volume) CFU / mL × 10 ⁷ TSST-1ng / ml TSST-1 ng / 10 ⁸ CFU % decrease 0 0 15.3 390 255 N / A 0 0.5 20.0 49 24 90% 0 0.25 24.6 24 10 96% 0 0.13 18.0 66 37 86% 0 0.06 18.3 124 68 73% 2.5 0 22.5 148 66 74% 2.5 0.5 1.5 3 19 93% 2.5 0.25 15.4 8 5 98% 2.5 0.13 26.2 47 18 93% 2.5 0.06 20.4 51 25 90% 5.0 0 25.6 203 79 69% 5.0 0.5 2.30 2 7 97% 5.0 0.25 12.0 7 6 98% 5.0 0.13 18.3 19 11 96% 5.0 0.06 26.2 28 11 96% 10.0 0 17.4 57 33 87% 10.0 0.5 6.1 2 4 99% 10.0 0.25 14.7 8 6 98% 10.0 0.13 20.0 16 8 97% 10.0 0.06 22.6 41 18 93% N / A not applicable

At all concentrations of cetiol 1414E, benzyl (s)-(-)-lactate increases TSST-1 production inhibition. The effect appears to be additive.

Example  5

In this example, the effect of benzyl laurate (Penta Manufacturing (Fairfield, New Jersey)) on the growth of Staphylococcus aureus MN8 and TSST-1 production was determined. The effect of the test compound was determined by placing the desired concentration, expressed in% (volume / volume), in 10 ml of growth medium as described in Example 1. Thereafter, the test compound was tested and evaluated as in Example 1. The effect of benzyl laurate on the growth of Staphylococcus aureus MN8 and TSST-1 production is shown in Table 7 below.

compound Amount of test compound (% (volume / volume)) Cetiol 1414E CFU / mL ELISA: TSST-1 ng / ml (% inhibition) none none 4.10E + 0.8 450 (N / A) Benzyl laurate 0.4% none 4.05E + 0.8 225 (62%) Benzyl laurate 0.6% none 3.80E + 0.8 55 (91%) none 10 mM 2.05E + 0.8 153 (79%) Benzyl laurate 0.4% 10 mM 2.05E + 0.8 60 (90%) Benzyl laurate 0.6% 10 mM 3.05E + 0.8 83 (86%) N / A = not applicable

At the tested concentrations, benzyl laurate reduced the amount of toxin produced by 62% to 91%. At the tested concentrations, benzyl laurate reduced the amount of toxin produced by 86% to 90% when combined with cethiol 1414E (mireth-3-myristate).

Example  6

In this example, the commercial tampons (superabsorbent KOTEX, Kimberly-Clark Worldwide, Inc. (Neenah, Wisconsin)) were treated with benzyl alcohol to effect the treated tampons on the growth and Staphylococcus aureus MN8 production and TSST-1 production. Was determined. As mentioned above, commercial tampons will contain about 0.75% (by weight of total tampons with cover) of Cetiol 1414E (Mireth-3-myristate). In preparation for the experiment, the cord of the tampon was cut off and the gauze was placed in a sterile 50 ml polystyrene test tube with the end of the cord down.

The gauze was inoculated with 5 ml of benzyl alcohol in five concentrations (0.0%, 0.3%, 0.15%, 0.075%, and 0.03%) dissolved in brain heart leach broth (BHI). Tampon gauze was left at room temperature for 1 hour.

Each gauze was then inoculated with 5.5 ml of inoculation broth containing 5 × 10 ⁶ ± 1 × 10 ⁶ CFU / ml Staphylococcus MN8 to achieve a final volume of 10.5 ml. The tube was blocked with a foam plug (IDENTI-PLUG plastic foam plug from Jaece Industries, purchased from VWR Scientific Products (South Plainfield, New Jersey)) and incubated at 37 ° C. for 24 hours. The gauze was removed from the incubator and individually placed in a sterile STOMACHER bag (Seward Ltd. (Norfolk, United Kingdom)) containing 50 ml sterile BHI. The gauze and fluid were then stommed or blended in the bag. Fluid aliquots were removed from the STOMACHER bag and placed in a sterile tube for testing.

Plate counting samples were prepared by vortexing the sample and collecting 5 ml of the sample and placing it in a new sterile 50 ml centrifuge tube. The samples were then sonicated at 8% power for 15 seconds using a Virsonic 600 ultrasonic cell mill (Virtis Company (Gardiner, New York)). After all samples were sonicated, the number of colony forming units (CFU) per ml was determined using a standard plate counting procedure.

In preparation for the analysis of TSST-1, the broth broth was centrifuged at 9000 rpm for 5 minutes at 4 ° C., and the supernatant was substantially filter sterilized through a 0.2 μm pore size 0.45 μm MCE filter. The resulting fluid was frozen at −70 ° C. in two 1 ml aliquots in a 1.5 ml polypropylene freezer vial with screw cap.

The amount of TSST-1 per ml was determined by non-competitive sandwich enzyme linked immunosorbent assay (ELISA). The method used was as follows: TSST-1 (# 606), rabbit polyclonal anti-TSST-1 IgG (LTI-101), rabbit polyclonal anti-TSST-1 IgG conjugated to horseradish peroxidase Four reagents of (# LTC-101), and normal rabbit serum (NRS) (# NRS-10), which were found to be free of anti-TSST-1, were prepared by Toxin Technology, Inc. (Sarasota, Florida). 62 μl of # LTI-101 was diluted so that 1: 100 dilution gave an absorbance of 0.4 at 205 nm, then added to 6.5 mL of Na ₂ CO ₃ buffer, pH 7.2, 0.5 M carbonate buffer, pH 9.6 , 100 μl of solution was pipetted into each inner well of a polystyrene microplate (available from Nunc-Denmark, catalog # 439454). The plate was covered and incubated overnight at 37 ° C.

Unbound anti-toxin was obtained from phosphate buffered saline (0.016 M Na ₂ HPO ₄ , available from Sigma Chemical Co. (St. Louis, Missouri)), pH 7.2, and 0.5% (volume / volume) Tween 20 (Sigma Chemical) Removed by washing four times in an automatic plate washer with 0.9% (weight / volume) NaCl (VWR Scientific Products (South Plainfield, New Jersey)) containing Co. (St. Louis, Missouri). Plates were treated with 100 μl of a 1% (weight / volume) solution of bovine serum albumin (BSA) fraction V (Sigma Chemical Co. (St. Louis, Missouri)) in Na ₂ CO ₃ + NaHCO ₃ buffer described above. The plate was covered again and incubated at 37 ° C. for 1 hour. Unbound BSA was removed by 6 washes with 250 μl PBS-twin.

The test sample was then treated with normal rabbit serum (10% (volume / volume) concentration) for 15 minutes at room temperature. TSST-1 reference standard (continuously diluted from 2-20 ng / ml in PBS-Twin) and NRS treated test samples (continuously diluted in PBS-Twin so that the resulting TSST-1 concentration is between 2-20 ng) Were pipetted into each well in a volume of 100 μl. Samples were then incubated at 37 ° C. for 2 hours, after which unbound toxins were removed by four washes with 250 μl PBS-twin. Rabbit polyclonal anti-TSST-1 IgG conjugated to horseradish peroxidase was diluted according to manufacturer's instructions. Final use dilution of the conjugates was determined by following the standard curve of the TSST-1 reference standard to undiluted conjugates, 1: 2 dilution conjugates, and 1: 4 dilution conjugates. Dilutions were chosen that gave results comparable to many existing conjugates. 100 μl of this dilution was added to each microtiter well. The plate was covered and incubated at 37 ° C. for 1 hour.

After incubation, the plates were washed six times with 250 μl PBS-twin. After washing, the wells were washed with 0.015 M sodium citrate, pH 4.0, 0.6 mM 2,2'-azino-bis- (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt and 0.009% (volume / volume) hydrogen peroxide (all 100 ml of horseradish peroxidase substrate solution consisting of Sigma Chemical Co. (available from St. Louis, Missouri). The intensity of the color response in each well was evaluated over time using VersaMax Molecular Devices microplate reader (OD 405 nm) and SoftMax Pro software (all available from Molecular Devices, Inc.). TSST-1 concentrations in the test samples were derived from the reference toxin regression equation for each assay procedure.

The efficacy of benzyl alcohol to inhibit TSST-1 production by Staphylococcus aureus is shown in Table 8 below.

Benzyl Alcohol Impregnation (wt%) CFU / mL (× 10 ⁸ ) TSST-1 ng / ml (% inhibition) 0 ^* 8.17 2.66 (N / A) 0.03% 6.13 2.46 (8%) 0.075% 2.83 2.22 (17%) 0.15% 0.5 1.33 (50%) N / A = not applicable * As mentioned above, commercial tampons are used in this embodiment. Thus, this sample contains about 0.75% (by weight of tampon with cover) of Cetiol 1414E (Mireth-3-myristate).

According to the present invention, the data shows TSST-1 in the presence of tampons containing both benzyl alcohol and cethiol 1414E (mireth-3-myristate) compared to control tampons containing Staphylococcus MN8 containing only cetiol 1414E. Indicates less production. At the tested concentrations, benzyl alcohol reduced the amount of toxin produced by 8% to 50%.

Example  7

In this example, the effect of growth of Staphylococcus MN8 on the integrity of various test compounds was determined by measuring the amount of test compound degradation caused by the enzymes produced by Staphylococcus MN8 bacteria. Desired concentrations of test compound were placed in 100 ml growth medium in 500 ml sterile Corning Fleaker (Fisher Scientific (Pittsbury, Pennsylvania)). Growth medium and inoculum were prepared as in Example 1.

Compounds to be tested include 0.2% (volume / volume) benzyl alcohol (Aldrich 40,283-4), 0.5% (weight / volume) sodium benzoate, 0.3% (volume / volume) benzyl (s)-(-)-lactate (Aldrich 42,484-6), 0.8% (volume / volume) benzyl ethyl malonate (Aldrich 30,069-1), and 0.3% (weight / volume) N-benzoyl-DL-leucine (Sigma B-1504). Test compounds were added to the growth medium in the amount necessary to obtain the desired final concentration.

Each flicker was inoculated with a determined amount of inoculum as described above. Flicker was blocked with sterile aluminum foil and incubated in atmospheric air at 35 ° C. in a Lab-Line orbital bath (available from VWR Scientific Products (McGaw Park, Illinois)) at 180 rpm. 15 ml samples were removed at 3, 6, 9, and 24 hours. The optical density of the cultures (595 nm) was determined and the cultures collected at 24 hours were analyzed for the number of colony forming units of Staphylococcus aureus MN8 using standard plate counting procedures. At the tested concentrations, the test compound did not inhibit the growth of Staphylococcus aureus.

In preparation for the integrity analysis of the test compounds, the cultures were centrifuged at 3000 rpm for 2 minutes at 2-10 ° C. Supernatants were filter sterilized through a 0.45 μm pore size, syringeless AUTOVIAL 5 filter (available from Whatman, Inc., Clifton, New Jersey). The resulting fluid was frozen in FISHERBRAND (12 mm × 75 mm) polystyrene culture tubes (Fisher Scientific (Pittsburgh, Pennsylvania)) at −70 ° C. until chemical analysis was performed.

Assays for 3, 6, 9, and 24 hours of undiluted solution were performed using Agilent Technologies 5973N GC / MS to assess the ability of Staphylococcus aureus MN8 to degrade test compounds. Based on the analysis, the control sample without the test compound added contained mostly acetic acid at all time intervals. Samples containing benzoic acid and benzyl alcohol were found to have benzoic acid and benzyl alcohol, respectively, as main compounds, but over time the benzoic acid and benzyl alcohol concentrations decreased. Samples containing N-benzoyl-DL-leucine as the test compound were found to contain benzoic acid as the main compound and the concentration decreased over time. Samples containing benzyl (s)-(-)-lactate as test compounds were found to contain benzyl alcohol as the main compound. It has also been found that the concentration of benzyl alcohol has increased over time. Finally, a sample containing benzyl ethyl malonate as the test compound was found to contain benzyl alcohol as the main compound. It has also been found that the concentration of benzyl alcohol has increased over time.

According to the present invention, the above GC / MS analysis indicates that the precursor compound was degraded by the enzyme produced by Staphylococcus aureus MN8 to produce the active species. It can also be seen that the precursor compound degrades slowly over time, allowing long-term inhibition of endocrine protein production by active species.

Further analysis of the 6 and 24 hour samples was performed by liquid chromatography. In contrast to chromatography, samples were diluted 10-fold with water. The dilutions were then analyzed using an ion exclusion column to achieve separation. Detection of the compound was carried out via UV absorption at 230 nm.

According to the present invention, liquid chromatography analysis of the test compound showed that the compound degraded into the active species benzoic acid and benzyl alcohol over a period of 24 hours. In particular, the 6 hour and 24 hour samples containing 0.3% N-benzoyl-DL-leucine showed evidence that the compound was degraded to benzoic acid. Additionally, 6 and 24 hour samples containing benzyl (s)-(-)-lactate and benzyl ethyl malonate showed evidence that the compound was degraded with benzyl alcohol. As can be seen further, the 24-hour sample containing benzyl (s)-(-)-lactate and benzyl ethyl malonate showed elevated levels of evidence of benzyl alcohol compared to the 6-hour sample.

In view of the foregoing, it will be presented that several objects of the invention are achieved and other advantageous results obtained.

In introducing the elements of the present invention or preferred embodiment (s) thereof, the indefinite article, the definite article, and “above” are intended to mean that one or more elements are present. The terms "comprising", "included" and "having" are intended to include and mean that there may be additional elements other than the listed elements.

As various changes may be made in the above without departing from the spirit of the invention, it is intended that all matter contained in the above and shown in the accompanying drawings shall be interpreted as illustrative and not in a limiting sense.

Claims (20)

As an endocrine protein inhibitor for inhibiting the production of exoproteins from Gram-positive bacteria, including non-absorbing substrates suitable for insertion into the vagina, an effective amount of a precursor compound of the formula Precursor compound inhibitors that can produce active species that are effective at inhibiting the production of endocrine proteins from Gram-positive bacteria upon hydrolysis:

[Wherein R ¹ is

And

It is selected from the group consisting of; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH. The endocrine protein inhibitor of claim 1, wherein R ⁵ is a monovalent saturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue having 1 to 15 carbon atoms, which may or may not be substituted with a hetero atom. The endocrine protein inhibitor of claim 1, wherein R ² is selected from the group consisting of H and OH, and R ³ and R ⁴ are independently H. ³ . The endocrine protein inhibitor of claim 1, further comprising a surfactant selected from the group consisting of mireth-3-myristate, glycerol monolaurate, and laureth-4. The process of claim 1 wherein the precursor compound consists of benzyl (s)-(-)-lactate, benzyl ethyl malonate, benzyl-laurate, benzyl benzoate, benzyl paraben, benzyl salicylate and phenoxyethyl paraben. Exocrine protein inhibitors selected from the group. The exocrine protein inhibitor of claim 1, wherein the precursor compound is present in an amount from about 0.15% (by weight of the non-absorbing substrate) to about 2.0% (by weight of the non-absorbing substrate). The endocrine protein inhibitor of claim 1, wherein R ⁶ is an amino acid and the amino acid is selected from the group consisting of valine, leucine and cysteine. The endocrine protein inhibitor of claim 1, wherein the precursor compound is selected from the group consisting of n-benzoyl-dl-valine, n-benzoyl-dl-leucine and n-benzoyl-dl-cysteine. The endocrine protein inhibitor of claim 1, wherein the non-absorbing substrate is selected from the group consisting of non-absorbable incontinence devices, barrier birth control devices, non-absorbable contraception devices, tampon applicators, and irrigation devices. 10. The endocrine protein inhibitor of claim 9, wherein the endocrine protein inhibitor is an irrigation apparatus comprising a vaginal cleansing formulation and the precursor compound is present in the vaginal cleansing formulation in an amount of about 0.2 mM / L to about 50 mM / L. An absorbent article for inhibiting the production of endocrine proteins from Gram-positive bacteria comprising an absorbent construct and an effective amount of a precursor compound of the formula: wherein the precursor compound is effective in inhibiting the production of endocrine proteins from Gram-positive bacteria upon hydrolysis. Absorbent articles that can produce species:

[Wherein R ¹ is

And

It is selected from the group consisting of; R ⁷ is -OCH _2- ; X is 0 or 1; R ⁵ is a monovalent saturated or unsaturated, substituted or unsubstituted, branched or straight chain hydrocarbyl residue, which may or may not be substituted by a substituted or unsubstituted aromatic ring, or a hetero atom; R ⁶ is selected from the group consisting of amino acids, methyl esters of amino acids, and ethyl esters of amino acids; R ² , R ³ and R ⁴ are independently selected from the group consisting of H, OH, COOH. 12. The absorbent article of claim 11 wherein R ⁵ is a monovalent saturated, substituted or unsubstituted, branched or straight chain hydrocarbyl moiety which may or may not be substituted with a hetero atom having 1 to 15 carbon atoms. The absorbent article of claim 11 wherein R ² is selected from the group consisting of H and OH, and R ³ and R ⁴ are independently H. 13. The absorbent article of claim 11 further comprising a surfactant selected from the group consisting of mireth-3-myristate, glycerol monolaurate and laureth-4. The compound of claim 11, wherein the precursor compound consists of benzyl (s)-(-)-lactate, benzyl ethyl malonate, benzyl-laurate, benzyl benzoate, benzyl paraben, benzyl salicylate, and phenoxyethyl paraben. An absorbent article selected from the group. The absorbent article of claim 11 wherein the precursor compound is present in an amount from about 0.15% (by weight of the absorbent structure) to about 2.0% (by weight of the absorbent structure). The absorbent article of claim 11 wherein R ⁶ is an amino acid and the amino acid is selected from the group consisting of valine, leucine and cysteine. The absorbent article of claim 11 wherein the precursor compound is selected from the group consisting of n-benzoyl-dl-valine, n-benzoyl-dl-leucine and n-benzoyl-dl-cysteine. The absorbent article of claim 11 wherein the absorbent article is selected from the group consisting of vaginal tampons, sanitary napkins, panty liners, incontinence underwear, contraceptive sponges, diapers, wound dressings, dental tampons, medical tampons, surgical tampons and nasal tampons. Absorbent supplies. The method of claim 19, wherein the absorbent article comprises a vaginal tampon comprising an absorbent tampon material and a cover material, wherein the precursor compound is at least about 2.6% (by weight of the cover material) to about 35% (of the cover material) Absorbent articles) by weight.