MXPA06010529A - Compositions comprising mannose phosphate for vaginal treatment - Google Patents

Abstract

The invention provides mannose phosphate and salts thereof for increasing vaginal cell growth, vaginal cell maturation and vaginal moisture, as well as compositions, articles and methods for treating and preventing vaginal conditions characterized by poor vaginal cell growth, low vaginal cell differentiation and low vaginal moisture.

Description

COMPOSITIONS COMPRISING MA OSA PHOSPHATE FOR VAGINAL TREATMENT Field of the Invention The invention relates to the use of mannose phosphate-containing compositions to promote maturation and proliferation of vaginal cells.

Background of the Invention Vaginal atrophy is a recognized and common problem in women with menopause. It is found that up to 50% of post-menopausal women as well as 10% to 20% in pre-menopausal women with low levels of estrogen. Vaginal atrophy is thought to be caused by low levels of estrogen resulting in a decrease in vaginal cell proliferation and differentiation. The decrease in cell proliferation leads to thinning of the vaginal epithelium and a lack of glycogen production by intermediate cells. Glycogen plays an important role in maintaining the vaginal ecosystem by serving as food for Lactobacilli acidophilus, the normal flora in the vagina, and by serving as a substrate for the production of acid to maintain a low vaginal pH. The thinning of the vaginal epithelium and the lack of glycogen in patients with vaginal atrophy frequently results in vaginal drying, discomfort and vaginitis.

Even though last-year replacement therapy has been used a bit of success in treating vaginal atrophy, new findings from the Women's Health Initiative indicate that such hormone replacement therapy increases the risk of heart attacks, strokes, of blood clots, and breast cancer.

Therefore, there is a need for non-toxic compositions and methods to treat vaginal atrophy without the need for hormone replacement therapies that may have negative side effects.

Synthesis of the Invention The present invention is directed to the composition and methods for treating and preventing vaginal conditions characterized by low vaginal cell proliferation, lower vaginal cell differentiation, lower vaginal moisture or vaginal atrophy. The composition and methods of the invention employ mannose phosphate, which can promote maturation and vaginal cell proliferation.

Therefore, in some embodiments, the invention is directed to a composition comprising an effective amount of mannose phosphate and a pharmaceutically acceptable carrier. The compositions of the invention can promote vaginal cell growth, vaginal cell maturity and can be used to increase the thickness and restore the health of the vaginal epithelium.

The invention is also directed to a method for treating or preventing a vaginal condition in a mammal. The method involves administering to a mammal an effective amount of a composition including a mannose phosphate or a salt thereof. As illustrated herein, mannose phosphate or salt thereof can increase the growth of mammalian epithelial cells and promote vaginal cell maturity. The compositions of the invention can therefore be used to treat conditions such as low vaginal cell proliferation, lower vaginal cell differentiation or lower vaginal moisture. In one embodiment, the condition is vaginal atrophy.

In one embodiment, the mannose phosphate employed is phosphate-6-mannose. As is known to one skilled in the art, phosphate-6-mannose can assume a number of conformations, all of which are contemplated by the invention. Some of the conformations that phosphate-6-mannose can assume are described below: where X is a double bond, hydrogen or a cation. The cation may be a monovalent or divalent cation, for example, sodium, potassium, than calcium, magnesium, manganese, zinc and the like.

Moreover, in some compositions of the invention some of the mannose phosphate molecules may have a variety of substituents in place of the hydroxy (-0H), aldehyde (-CH0), and hydrogen substituents that are generally found in mannose phosphate preparations. In general, the exact type of substituents in the mannose phosphate composition (s) of the invention can be varied to stimulate optimal levels of vaginal cell growth and / or maturity. Also contemplated are the phosphate-6-mannose polymers that can be broken down into mannose phosphate monomers. Such polymers can include other saccharides or non-saccharides that break to allow the release of mannose phosphate.

The compositions of the invention are generally administered intravaginally. However, other routes are also contemplated. For example, the compositions may also be administered topically. The compositions can be incorporated into feminine products such as washes, tampons, foams, creams, sustained-release implants and the like for easy use and administration. The invention further provides similar applicators to the syringe for the administration of the compositions of the invention. Such applicators may contain a mannose phosphate composition of the invention. Washes, buffers, foams, creams, sustained release implants and applicators can be prepared in a sterile manner to optimize shelf life of the composition to allow the composition to be delivered in a sterile manner.

An effective amount of the compounds of the invention may vary, but in some embodiments the effective amount of mannose phosphate may be in the range of about 0.01 micrograms to about 500 milligrams.

Brief Description of the Drawings Figure 1 illustrates the effect of phosphate-6-mannose (M6P) on cell proliferation in normal human vaginal epithelial cells in culture medium.

Six samples were tested in each group; the bar error represents the standard deviation.

Figure 2 illustrates the treatment of phosphate-6-mannose (M6P) increases the observed numbers of mature vaginal epithelial cells as measured by the production of glycogen. The production of glycogen in the vaginal epithelial cell cytoplasm is a marker of differentiation / maturity for vaginal epithelial cells. Each group had 5 samples and the bar error represents the standard deviation.

Figure 3 is a schematic diagram of a type of applicator similar to the syringe that can be used to deliver a mannose phosphate composition to a vagina of a mammal. The syringe-like applicator consists of a barrel 20 and a plunger 30 with a plunger head 40. The syringe-like applicator may also have a barrier seal 60 distal to the plunger head 40. The applicator similar to the The syringe may also have a chamber 50 within the barrel resting on the plunger 30 and the barrier seal 60. The chamber 50 comprises an effective amount of a mannose phosphate compound or a salt thereof. The presence of the barrier seal 60 seals the applicator and maintains the composition contained with the applicator during shipping and handling. The barrier seal 60 can be removed by the user, or it can be broken when the user depresses the plunger. At the time of use, the applicator is inserted into the vagina and the plunger 30 is depressed. This force can push the composition out of the applicator and into the vagina.

Figure 4 is a schematic diagram of a vaginal insert that can be used to deliver a mammalian phosphate composition to the vaginal of a mammal. The vaginal consists of a tubular shaped material 110 comprising a mannose phosphate compound and a cord 120 or other union for the recovery or positioning of the vaginal insert within the vagina. In some embodiments, cord 120 is optional, for example, because the vaginal insert slowly dissolves, breaks or corrodes to provide sustained release of mannose phosphate. Therefore, recovery may not be necessary. The syringe-like applicator illustrated in Figure 3 can be used for the delivery of the vaginal insert.

Detailed description of the invention The invention provides compositions and methods for increasing vaginal cell proliferation and vaginal cell maturity or differentiation. Such compositions can promote vaginal cell growth, the development of differentiated vaginal cells and can be used to increase the thickness and restore the health of the vaginal epithelium. The compositions employed by the invention are inexpensive, non-toxic and readily available. The effectiveness of these compositions and methods does not depend on hormone replacement or substances that are not naturally found in the body. Therefore, the composition avoids the negative side effects associated with commonly used hormone replacement therapies.

Compositions of Phosphate of Mañosa The compositions employed by the invention to increase vaginal cell proliferation and maturity contain a mannose phosphate, preferably phosphate-6-mannose (M6P).

In some embodiments, other saccharides may also be included in the compositions of the invention. For example, mannose phosphate compositions may include glucose-1-phosphate, glucose-6-phosphate, mannose-1-phosphate, galactose-6-phosphate, fructose-6-phosphate, glucose-1,6-diphosphate, or fructose-1, 6-diphosphate. The percentage of alternating saccharides in the compositions of the invention may vary. For example, the compositions of the invention may include 0% up to about 50% alternating saccharides. In other embodiments, the compositions of the invention may include from about 0% up to about 40%, or 0% up to about 30%, or 0% up to about 20%, or 0% up to about 10% alternating saccharides .

Moreover, the mannose phosphate for use in the present invention can be formulated as pharmaceutically acceptable salts or cosmetically thereof, for example, mono- or disodium salts, as well as any precursor forms that when applied to the epithelium or to the Skin releases the mannose phosphate.

In some embodiments of the sugar cane phosphate units may have a variety of substituents in place of the hydroxy (-OH), the aldehyde (-CHO), and hydrogen substituents that are typically found in the mannose phosphate. For example, the lower alkyl moieties can replace any of the hydrogen atoms of the hydroxy (-OH) or hydrogen substituents of the mannose phosphate compounds employed in the invention. The lower alkyl amino or amino groups can replace any of the OH or hydrogen groups of the mannose phosphate compounds employed in the invention. The sulfate (S0) can replace the phosphate groups of the mannose phosphate compounds employed in the invention. Therefore, substituents that may be present instead of, or in addition to, substituents typically present in the mannose phosphate compounds include sulfate (S04), lower alkoxy, lower alkanoyloxy, and / or lower alkanoylaminoalkyl.

As used here, lower alkyl means (Ci-Cß) alkyl. Tal. { C? -Cβ) alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl. Preferred lower alkyl groups are (C? -C3) alkyl including methyl ethyl, propyl, isopropyl and the like. Lower alkoxy means C? -C6) alkoxy; such (C6C6) alkoxy may, for example, be methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-bitoxy, pentoxy, 3-pentoxy, or hexyloxy. The hydroxy lower alkyl refers to a hydroxy group attached to a lower alkyl or lower alkylene group (for example -CH2-CH2-OH). The lower alkanoyloxy refers to (C2-C6) alkanoyloxy, for example, acetoxy, propanoyloxy, butanoyloxy, isobutanoyloxy, pentanoyloxy, or hexanoyloxy. The lower (Ci-Cg) alkanoylamino may, for example, be acetamino, propanoylamino, butanoylamino, isobutanoylamino, pentanoylamino, or hexanoylamino.

Therefore, while the active ingredients in the compositions of the invention typically include a high percentage of mannose phosphate some variability in the types of substituents and sugar units present in the mannose phosphate preparation employed is acceptable as long as the preparation can increase vaginal cell maturity and proliferation.

In other diments, the polymers and mixtures of mannose phosphate with other polymers can be employed in the compositions and methods of the invention. Polymer phosphate polymers that can be broken to release mannose phosphate monomers are particularly desirable. In some diments, the non-saccharide polymers can be used in the mannose phosphate formulations. Examples of polymers that can be used with mannose phosphate include proteoglycans, poly (ethylene glycol) / (poly (ethylene oxide), poly (vinyl alcohol), poly (vinylpyrrolidone), and poly (2-) hydroxylethyl methacrylate) Such polymers can be covalently bound to the mannose phosphate or simply combined with mannose phosphate to form a mixed composition.

Methods of Use The present invention is directed to methods of treating or preventing or otherwise improving vaginal conditions characterized by poor vaginal cell growth, lower vaginal moisture and poor vaginal cell differentiation. The compositions of the invention can also be used in vaginal moisturizers and in methods to moisturize the vagina and relieve vaginal dryness and restore health to the vaginal epithelium.

In some diments, the compositions and methods of the invention are used in subjects with an abnormally thin vaginal lining or in subjects with an abnormally thin vaginal mucosa. The symptoms that result • Abnormally thin mucosal or vaginal lining include vaginal dryness, discomfort, itching, dyspareunia, infection, inflammation, ulcers, discharge, and bleeding. In some instances, the compositions and methods of the invention are used to treat vaginal atrophy.

Vaginal atrophy is a condition that occurs in some women, typically in postmenopausal women, in whom there is a significant thinning of the vaginal mucosa. The thin vaginal mucosa lacks maturity, which means consists of numerous parabasal and few or non-superficial cells and intermediate cells, resulting in decreased glycogen deposits and a higher pH. Vaginal atrophy is mainly caused by an estrogen deficiency because the vaginal mucosa is a tissue sensitive to estrogen and a well-known organ or target for estrogen. As provided herein, vaginal atrophy and the effects of inferior estrogen in the vagina can be coated by treatment with the compositions and methods of the invention.

These methods include administering to the animal an effective amount of mannose phosphate, for example, a therapeutically effective amount of mannose phosphate. According to the invention, a therapeutically effective amount of mannose phosphate can increase vaginal epithelial cell growth, increase the percentage of differentiated or mature vaginal epithelial cells and increase vaginal moisture.

The treatment of, or treating, a vaginal condition is intended to include the alleviation of or abatement of at least one symptom typically associated with the vaginal condition, for example, at least one symptom typically associated with vaginal atrophy. Treatment also includes alleviating or decreasing more than one symptom. Ideally, treatment cures, for example, substantially eliminate one or more symptoms associated with the vaginal condition.

Administration According to the invention, the mannose phosphate compounds, polymers, copolymers, polymer blends and their salts, can promote vaginal cell proliferation and / or vaginal cell maturity in animals. The term "animal", and as it is used here, generally refers to a warm-blooded animal. Mammals include cattle, buffalo, sheep, goats, pigs, horses, dogs, cats, rats, mice, rabbits, chickens, turkeys, and humans. Also included are other cattle, domesticated animals and captive animals.

An effective amount of a mannose phosphate compound, polymer, copolymer, polymer mixture and salts thereof, to promote vaginal cell growth and / or maturity is an amount that increases the growth of a population of vaginal cells, by example, the epithelial cells, relative to a population of the same type of vaginal cells that did not receive mannose phosphate. To achieve the desired inhibition, the composition can be administered as a single or divided dose, for example, of at least about 0.001 μg to about 100 to 200 milligrams, from about 0.01 μg to about 75 to 100 milligrams, from about 0.1 μg to about 50 to 75 milligrams or about 1.0 μg to about 30 to about 50 milligrams of a no or more phosphate-6-mannose compound, although other doses may provide beneficial results. In some embodiments, the dose may vary from about 0.01 milligram to about 50 milligrams.

Daily doses of the compositions of the invention may also vary. Such daily doses may be in the range, for example, of about 0.001 milligrams per day to about 500 milligrams per day, from about 0.01 milligrams per day to about 250 milligrams per day, from about 0.01 milligrams per day to about 120 milligrams per day, from about 0.1 milligrams per day to about 100 milligrams per day, from about 0.1 milligrams per day to about 75 milligrams per day, and from about 0.1 milligrams per day to about 50 milligrams per day of one or more mannose phosphate compounds.

The amount of administered may vary depending on several factors including, but not limited to, the disease, weight, physical condition, health, age of the animal, and whether the prevention or treatment of vaginal atrophy can be achieved . Such factors can be easily determined by the clinician who employs animal models or other test systems are available in the art.

The compositions of the invention can be administered for prophylactic, therapeutic, and / or hygienic use. Such administration can be topical, vaginal, rectal, and other convenient routes. The administration can be directly to the epithelial cell surfaces. For example, the compositions of the invention can be administered directly to mucosal surfaces. The mucosal surfaces include the vaginal, the urogenital, the rectal and the like. The surfaces of the urogenital tract that can be treated with the compositions and methods of the invention include rectal, urethral, ureteral, vaginal, cervical, uterine, etc. In many embodiments, the epithelial cell surface is vaginal .

The administration of therapeutic agents according to the present invention may be in a single dose, in multiple doses, in a continuous or intermittent manner, depending, for example, on the physiological condition of the container, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to practitioners with skill. For the prevention of certain conditions or diseases (for example vaginal atrophy), the administration of the compositions of the invention can be essentially continuous over an indeterminate period of time, for example, at regular intervals for life. Alternatively, the compositions of the invention can be continuously administered for a preselected period of time or in a series of spaced doses. The local administration is generally contemplated.

The compositions are prepared by combining active ingredients in the appropriate concentrations. Other active or inactive agents selected by one of skill in the art may optionally be added. The absolute weight for an active agent in a unit dose can vary widely.

The compositions of the invention may be administered in the form of an article or carrier such as a vaginal insert, or by means of a syringe-like applicator, a tablet, a suppository, a pessary, a powder / talc or other solid, a solution, a liquid, a sprayer, an aerosol, a wash, an ointment, a tampon, a foam, a cream, a gel, a paste, a microcapsule (s), a vaginal sponge, a vaginal ring, a controlled release formulation, a sustained release formulation or a bioadhesive gel (e.g., a mucoadhesive thermogelant composition (see, e.g., U.S. Application No. 10/135805, filed April 30, 2002 , which is incorporated herein by reference)).

For intravaginal administration, therapeutic agents can be formulated as is known in the art for direct application in the vaginal area. Forms mainly conditioned for vaginal application take the form, for example, of creams, milks, gels, dispersions, micro-emulsions, thick lotions to a greater or lesser extent, impregnated pads, ointments, aerosol formulations ( for example, sprays or foams), creams, lotions, pastes, jellies, sprays, and sprays. Alternatively, the composition can be formulated to be part of an adhesive polymer, such as a polyacrylate or an acrylate / vinyl acetate copolymer.

The ointments and creams can, for example, be formulated with an aqueous or oily base with the addition of thickening agents and / or gelatins. The lotions may be formulated with an aqueous or oily base and may in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents or coloring agents. Liquid sprays are conveniently supplied with pressurized packaging, for example, by means of a specially formed enclosure. The active compositions can also be delivered by means of iontophoresis, for example, as described in US Pat. Nos. 4,140,122; 4,383,529; or 4,051,842. The percentage by weight of a prophylactic agent of the invention present in a formulation may depend on several factors, but can generally be about 0.01% to about 98% of the total weight of the formulation, and typically about 0.1% up to around 90% by weight.

The pharmaceutical formulations of the present invention may include, as optional ingredients, pharmaceutically acceptable carriers, diluents, solubilizing or emulsifying agents, and salts of the type that are available in the art. Examples of such substances include normal saline solutions such as physiologically quenching saline solutions and water. Specific non-limiting examples of carriers and / or diluents that are useful in the pharmaceutical formulations of the present invention include water and physiologically acceptable salt buffer solutions such as phosphate buffer saline solutions with a pH of about 4.5 to about 5.5 Additionally, the active ingredients may also be used in combination with other therapeutic agents, for example, antimicrobial agents, pain pills, anti-inflammatory agents, vitamins (e.g. vitamin B, C or E), aloe vera and the like, either for conditions described here or some other condition. Previous work by the inventors shown in the hyaluronic acid compositions can inhibit pathogen binding to a variety of cell types, including vaginal epithelial cells. See U.S. Patent No. 10 / 401,522 and U.S. Patent No. Serial No. 10 / 608,848, which are incorporated herein by reference. Moreover, the inventors have shown that hyaluronic acid can promote vaginal cell maturity and growth. See U.S. Patent No. Serial No. 10 / 696,547, which is incorporated herein by reference. Therefore, according to the invention, compositions containing mannose phosphate may also contain hyaluronic acid.

The present invention also concerns a pharmaceutically packaged composition for controlling or preventing a vaginal condition (e.g. vaginal atrophy) such as a case or other container. The kit or container maintains a therapeutically effective amount of a pharmaceutical composition for preventing, controlling or inhibiting the vaginal condition and instructions for using the pharmaceutical composition for the prevention, control or inhibition of the vaginal condition. The pharmaceutical composition includes mannose phosphate, in a therapeutically effective amount such that the vaginal condition is prevented, controlled or inhibited.

Additionally, the invention provides a vaginal insert that can release the mannose phosphate in a controlled manner. Such a vaginal insert may be biodegradable or non-biodegradable. The vaginal insert provides sustained release of the active ingredients at an appropriate rate to achieve the desired degree of vaginal cell growth or vaginal cell maturity, or the desired degree of treatment or prevention of vaginal atrophy.

In some embodiments, the active ingredients may be formulated with ointments or oleaginous bases to form a semisolid composition with a desired shape. For example, the composition can be formed for easy insertion into a hole such as the vagina. This class of formulations comprises the active ingredients and the hydrocarbon-based semi-solids containing dissolved and / or suspended bacteriostats / preservatives and a buffer system. The petrolatum component in these bases can be any paraffin in the viscosity range of mineral oil using incorporated isobutylene, colloidal silica, or stearate salts to paraffin waxes. The white and yellow petrolatums are examples of such petrolatum components. Bases of this kind can be made by incorporating top cast waxes into a fluid mineral oil by means of melting or by incorporating polyethylene in mineral oil at elevated temperature. Polysiloxanes (also known as silicones) are suitable for use in these bases and typically have a viscosity in the range of about 0.5 to 106 centistokes. The organic entities attached to the polysiloxane are preferably lower molecular weight hydrocarbon moieties having from 1 to 8 carbons each, such as lower alkyl, lower alkenyl, phenyl and substituted alkyl phenyl, and lower alkyl phenyl, such as the benzyl. In such half, each lower alkyl or alkenyl group preferably has 1 to 3 carbons inclusive, such as in a dimethylsiloxane polymer.

Absorption bases can be used with such oleaginous system. In addition to the active ingredients, additional ingredients with the ability to emulsify a significant amount of water are employed. Water-in-oil demolitions (w / o) can be formed where the external phase is oily in character. Condoms / bacteriostats, such as parabens, shock absorber systems, etc. they can be incorporated in these bases as emulsified aqueous solutions together with the active ingredient. Various additives are conveniently used as the emulsifier, and these include, but are not limited to cholesterol, lanolin (which contain cholesterol and cholesterol esters and other emulsifiers), lanolin derivatives, beeswax, fatty alcohols , wool wax alcohols, HLB emulsifiers (hydrophobic / lipophobic balance), and assorted ionic and nonionic surfactants, singly or in combination.

The water-in-oil emulsifier bases can be used in the compositions, inserts and articles of the invention. These formulations may be an expansion of the general class of absorption bases that include liquids or creams. These can be prepared by taking a mixture of the active ingredients with oil phase ingredients, bacteriostats / preservatives and buffer salts that are dissolved or suspended therein and to which water has been added to form a water-in-oil emulsion.

The oil-in-water emulsion bases can also be used in the compositions, in the inserts and in the articles of the invention. These systems are semisolid emulsions, microemulisions, or foam emulsion systems. Usually such a system has a "creamy white" appearance. Typically, the internal oil phase is in the range in percent of the composition of about 10% to about 40% oil by weight and the external phase may contain 80% or more of water. The oil phase may contain, but is not limited to, long chain alcohols (cetyl, stearyl), long chain esters (myristates, palmitates, stearates), long chain acids (palmitic, stearic), vegetable oils and animals and assorted waxes. These can be made with anionic, cationic, non-ionic or amphoteric surfactants, or with combinations especially of non-ionic surfactants.

The inserts and suppositories containing the active ingredients can be, for example, oleaginous in nature that melt at body temperature, or polyethylene glycol-based compositions that dissolve in mucosal fluids (eg, vaginal). Additional bases for suppositories are glycerin and glycerinated gelatin.

The active ingredients can be formulated in inserts, in articles, in tampons, in transdermal patches, bandages, and bandages using cushioned gels made with gelling agents. Some examples of these gelling agents are: cellulosics, cationic polymers, polyoxyalkylenes, and carboxyvinyl polymers. Cellulosics useful in the formulations of the invention ine, for example, methyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose. Cationic polymers useful in the formulations of the invention ine "Polyquaternium-10", a polymeric quaternary ammonium salt of hydroxylethyl cellulose reactivated with a substituted trimethyl ammonium epoxide, and the like. Polyoxyalkylenes useful in the invention ine the polyoxypropylene polyoxyethylene esters of lanolin and the derivatives thereof. Carboxyvinyl polymers useful for the formulations of the invention ine the crosslinked acrylic acid polymers, for example, those commercially available from B.F. Goodrich Co., Akron, Ohio, under the designation of CARBOPOL ™.

Controlled or sustained release can be achieved by the addition of time release additives, such as matrices, of polymeric structures, which are available in the art. For example, the compositions of the invention may also be administered through the use of hot melt extrusion articles, such as the interlaced hot-melt extruded film (see, for example, U.S. Patent No. 6,375,963, which is incorporated herein by reference). The formulation may comprise an interlaced polycarboxylic acid polymer formulation, generally described in United States of America Patent No. 4,615,697 issued to Robinson (hereinafter "the 697 patent"), which is incorporated herein by reference. In general, about 80% of the polymer monomers in such formulation contain at least one carboxyl functionality. The entangled agent must be present in such an amount in order to provide sufficient adhesion to allow the system to remain attached to the surfaces of the epithelial or endothelial cells for a sufficient time to allow the desired release of mannose phosphate to take place.

An insert or article may comprise a mixture of polymers that provide release of the active agents at a constant rate over a prolonged period of time. In some embodiments, the article or insert comprises water-soluble pore-forming agents, such as polyethylene glycol (PEG) that can be mixed with water-soluble polymers to increase the durability of the insert and to prolong the release of the active ingredients. . Such a water-soluble pore-forming agent may be polyethylene glycol, polypropylene glycol, a mixture or sugars of Lime po (lactose, sucrose, dextrose, etc.), salts, poloxamers, hydroxypropyl cellulose, polyvinyl alcohol and other soluble food grade in water and other excipients.

When polyethylene glycol is used as a pore forming agent, the molecular weight of polyethylene glycol is in the range of about 200 to about 20,000, alternatively, from about 400 to about 8,000. For example, polyethylene glycol having a molecular weight of about 540 to about 8,000 is used. In another embodiment, the polyethylene glycol has a molecular weight of about or above 1,000 to about 8,000. The molecular weight of the polyethylene glycol used for the coating with the formulation of the invention may depend on the ability of the polyethylene glycol to form a coating film that is non-tacky, has sufficient strength and creates an adequate pore size to control the release of the active ingredients over the desired period of time both in vitro and in vivo.

The pore forming agent is used in the formulation of the invention in the amount effective to regulate the release of a mannose phosphate compound at a desired rate. Preferably, the effective amount of the pore forming agent provides long-term supply of the active ingredient thereby increasing the shelf life of a sustained release insert, article or implant. The effective amount of the pore-forming agent may depend on the desired rate and duration of release and the ability to form a continuous microporous film during the coating process. To allow the duration of release over longer periods of time polypropylene glycol with higher molecular weights is used. For example, polyethylene glycol 8000 can provide release over a period of time that is longer than 100 days, when it is used in a concentration of 10% to 50%, preferably 20% to 45% and more preferably 30% to Four. Five%. The polyethylene glycol concentration is expressed here in% by weight per dry basis and represents the concentration of polyethylene glycol in the coating film after drying. Similarly, the thickness of the coating film is from 5 to 50 μm, preferably 30 from 10 to 30 μm and more preferably from 15 to 25 μm.

There is a good correlation between the dissolution rate of active agents and the amount of pore forming agent incorporated in the coating film based on in vitro and in vivo studies. Depending on the desired length of release, the concentration ranges of the polyethylene glycol can be adjusted as needed. For example, the in vivo duration of a coated insert can simply be predicted from the in vitro dissolution rate of the active agent at the 120-hour time point.

The inserts and articles of the invention may also comprise a water insoluble polymer. Examples of such polymers are ethylcellulose, acrylic resins, methacrylic acid copolymer and ethyl ester of acrylic acid, polylactic acid, PLGA, polyurethane, polyethylene vinyl acetate copolymer, polystyrene copolymer. butadiene and silicone rubber, or mixtures thereof. For example, polymers sold under the names Aquacoat ECD 30 and Eudragit RS 30 and NE 30D (trademarks of Rhom Tech, Inc.) may be used.

A polymer suitable for use in this invention is a polymer that can release active ingredients of the invention at a rate that is sufficient for the treatment of a vaginal condition described herein. The rate of the control formulation prepared with such a polymer is stable during implantation. The formulation should have sufficient strength to withstand routine handling, and have the stability to release the active ingredients at an acceptable rate.

In one embodiment, the coating formulation of the invention is used to coat pellets comprising the active ingredients that are compressed to form a biodegradable, solid insert and then administered to promote proliferation and vaginal cell maturity.

A polymer formulation can be used to provide controlled or sustained release. Such a polymer formulation can be adjusted to control the release rate of the mannose phosphate by varying the amount of interlacing agent in the polymer. Suitable entangled agents include divinyl glycol, divinylbenzene, N, N-diallyl acrylamide, 3,4-dihydroxy-l, 5-hexadiene, 2,5-dimethyl-l, 5-hexadiene and the like.

An example of a polymer for use in such a formulation is Polycarbophil, U.S.P., which is commercially available from B.F. Goodrich Specialty Polymers of Cleveland, Ohio under the brand designation NOVEON ™ -Al. The United States Pharmacopeia, 1995 edition, United States Pharmacopeial Convention, Inc., Rockville, Meriland, at pages 1240-41, indicates that polycarbophil is a polyacrylic acid, entangled with divinyl glycol.

Other useful bioadhesive polymers that can be used in such a drug delivery system formulation are mentioned in U.S. Patent No. 4,615,697. For example, these include polyacrylic acid polymers entangled with, for example, 3,4-dihydroxy-l, 5-hexadiene, and polymethacrylic acid polymers entangled with, for example, divinyl benzene. Typically, these polymers can not be used in their salt form, because this may decrease their bioadhesive capacity. Such bioadhesive polymers can be prepared by conventional free radical polymerization techniques using initiators such as benzoyl peroxide, azobisisobutyronitrile, and the like. Exemplary preparations of useful bioadhesives are provided in the '697 patent.

For vaginal administration, the formulation preferably degrades slowly or remains attached to the epithelial or endothelial cell surfaces for a period of at least one to forty-eight hours. Such results can be clinically measured over several time periods, by testing the vagina samples for mannose phosphate. The bioadhesion of a formulation of the invention can be achieved with bioadhesive polymers using an entangled agent that is present at about 0.1% to 6.0% by weight of the polymer, with about 1.0% to 2.0% by weight in some embodiments, for achieve an appropriate level of bioadhesion. Bioadhesion can also be measured by commercially available surface tensiometers used to measure adhesive strength.

The formulation can be in the form of a gel, a cream, a tablet, a capsule, a suppository, a film, or any other pharmaceutically acceptable form that is tolerated by epithelial cells (e.g., the mucosa) and is not washed so easily. Different formulations are additionally described in U.S. Patent No. 4,615,697, which is incorporated herein by reference.

As will be apparent to those skilled in the art, the composition of the formulation can be varied to affect certain properties of the formulation. For example, the viscosity can be varied by adding a polymer or gel former. In some embodiments, a bioadhesive polymer can be included in various concentrations to provide more or less bioadhesion. A pH-sensitive bioadhesive can be used to effect greater release at certain pH values. The particular bioadhesive qualities should prevent the composition from being diluted or washed, thus increasing the usefulness of the present formulation.

The liquid compositions of the invention can be administered from absorbent materials, such as a bandage, a tampon or a sponge, or as a spray / aerosol (applied to the affected area using a spray type pump or aerosol). The use of a buffer, in which the composition of the invention has been incorporated, it is advantageous in that the composition may be slowly or continuously released even though it may be continuously transported away by menstrual blood or other vaginal discharge. Provide the composition in the form of a solution, which may be initially supplied in a concentrated liquid form, or as a dissolvable powder, a tablet or the like which requires the addition of water, saline or other suitable diluents before its use, allows the composition to be administered as a vaginal wash.

The solid compositions of the invention can be applied by any number of means, including the use of applicators or by self-insertion by the patient. For example, creams, lotions, suppositories, foams, pastes, ointments, gels, tablets, or buffers can be administered using an applicator, such as a plunger type or plunger type applicator. Administering the composition as a vaginal suppository is advantageous as it provides increased convenience, ease of application, safety and / or cleanliness. Administering the composition as a cream having a lower surface tension is advantageous since it provides a uniform wetting action which aids in the penetration of the composition into crypts and crevices of the orifice. Such creamy composition can also act as a humectant.

Additionally, the additives may be mixed with the formulation for maximum or desired efficacy of the delivery system or for the comfort of the patient. Such additives include, for example, lubricants, plasticizing agents, preservatives, gel formers, tablet formers, pill formers, suppository formers, film formers, cream formers, de-inking agents, coatings, binders, vehicles, coloring agents, agents that control taste and / or odor, humectants, viscosity controlling agents, pH adjusting agents, and the like agents.

A desired embodiment provides for compositions of the invention in a syringe-like applicator (also known as a plunger-type applicator or similar to the syringe (see Figure 3)). For example, a composition including mannose phosphate can be applied in a chamber 50 with a barrel 20 of an applicator similar to the syringe. The chamber is sealed at the distal end with a barrier seal 60 and the proximal end by the plunger head 40. The presence of the barrier seal 60 seals the applicator and maintains the composition contained with the applicator during shipping and handling. However, the barrier seal 60 may be removed by the user or may be broken when the user depresses the plunger 30. At the time of use, the applicator is inserted into the vagina and the plunger 30 is depressed. This force can push the composition out of the applicator and into the vagina. As an alternative, a tapered tip can be used in place of the barrier seal 60.

An embodiment of the invention provides an aqueous gel containing a mucoadhesive material, such as carboxymethylcellulose (optionally mixed with a mucoadhesive thermo-freezing agent), to be mixed with mannose phosphate to thereby form a composition of the invention. A further incorporation provides for the encapsulation of mannose phosphate into polymeric microparticles. Once in place, the polymer dissolves and the mannose phosphate is released. In this case, the release of the phosphate from the stone can be controlled by the microparticles to provide the extended production of the desired product. (for example, sustained release). The delivery vehicle is not limited to use in the vagina, but can also be applied to a wide variety of biomedical applications where the supply of mannose phosphate is desired. The proper modification of the delivery vehicles described herein is within the skill of those in the art.

Additionally, the composition and / or delivery materials may contain additional beneficial agents that may improve the health of the vagina. For example, polymers used as carriers or for encapsulation or for sustained release can be hydrolytically degraded in an acid or acid producing species. One of such polymer is a poly (vinyl alcohol) backbone with pendant polycaprolactone chains that, upon disintegration, yields polyvinyl (polycaprolactate). Polycaprolactone is hydrolytically degraded into a caproic acid. This acid helps in lowering the pH and controlling harmful bacterial growth, so it helps restore the balance to the epithelium. Additionally, this material is processable melt and can be formed in a system for the controlled supply of the mannose phosphate, additionally, a Laureth-4 peroxide (for example, a Laureth-4 terminal peroxide) will be able to release laureth-4 and peroxide ( for example, hydrogen peroxide). The laureth-4 decreases the production of TSS-1 by S. aureus and the peroxide are available to suppress anaerobes and Gardnerella vaginalis, so it reduces the production of toxins while the vaginal flora is restored.

The examples further illustrate certain aspects of the invention and are intended to limit the invention in any way.

EXAMPLE: Phosphate-6-Mannose Promotes Proliferation and Vaginal Cell Maturity This example describes experiments that show that mannose phosphates can promote vaginal cell proliferation and vaginal cell maturity.

Materials and methods Clonetics normal human vaginal epithelial cells (NHVE 5164) were subcultured in basal PrEBM (Clonetics CC 3165) into 96 well plates at 37 ° C, 5% C02. The cells were then exposed to a medium containing mannose-6-phosphate (Sigma, M3655) at a concentration of 10"5 M. The control groups received a culture medium without mannose-6-phosphate. examined three days later and glycogen positive cells were detected five days after treatment with mannose-6-phosphate.

Cell proliferation was examined by using a solution of CellTiter 96 Aqueous One from Promega (# TB245). Twenty μl of the reagent were added to each well.

The plates were returned to the cell culture incubator for 3 hours. The absorbance in each well was measured at 490 nm with a micro-concentration reader.

A periodic acid Schiff assay (PAS) was used to detect glycogen levels. Before the assay, the cells were washed with PBS and fixed with a 10% by volume solution of Formalin / ethanol for 1 hour at room temperature, followed by a continuous rinse with deionized water (DI) for 1 minute. A Harleco® PAS assay kit (EM Science 64945/43) was used for the detection of glycogen. The basic principle is that the hydroxy groups on the glycogen units were first oxidized in aldehyde groups by periodic acid, and the aldehyde groups reacted with a Schiff agent to produce a red dye. The specific assay operations were all done at room temperature as follows: 1. The periodic acid reagent was added to the well / chamber and allowed to remain for 10 minutes. 2. The samples were continuously rinsed in water for one minute. 3. The Schiff reagent was added and the samples were allowed to remain for 15 minutes. 4. The samples were continuously rinsed with deionized water for one minute.

. The sodium carbonate reagent (2X diluted with deionized water) was added and the samples allowed to remain for 5 minutes. 6. The samples were continuously rinsed in deionized water for 5 minutes. 7. The yellowish green light SF reagent was added and the samples were allowed to stand for 30 seconds. 8. The samples were rinsed in deionized water briefly for 10 seconds. 9. The samples were dehydrated with a series of ethanol / water mixtures (75%, 90%, 95% to 100% ethanol) and xylene.

. The samples were assembled with a mounting medium.

The numbers of positive cells stained with glycogen were manually counted under a microscope. The data was analyzed by the student test t and p < 0.05 was considered significant.

Results As shown in Figure 1, the mannose-6-phosphate treatment significantly increased (p <0.05) vaginal cell proliferation. In addition, the number of glycogen-positive cells was significantly higher than the vaginal cells that were treated with ma-6-phosphate (figure 2).

These results indicate that mannose-6-phosphate can stimulate cell proliferation and vaginal cell maturation. Vaginal atrophy is the thinning of the vaginal epithelium that can be caused by low estrogen levels that result in a decrease in vaginal cell proliferation. Atrophic vaginitis, the overgrowth of pathogens due to the thinning of the protective epithelium, can develop in patients with vaginal atrophy. Therefore, compositions containing mannose-6-phosphate that stimulate vaginal cell proliferation can be used to increase the thickness of the vaginal epithelium, stimulate the maturation of differentiated vaginal cells and restore healthy vaginal epithelium.

All of the patents and publications mentioned herein are indicative of the skill levels of those skilled in the art to which the invention pertains, and each of said patents applies mentioned publications herein incorporated by reference to the same extent that it would have been. incorporated by reference in its entirety individually or as set forth herein in its entirety. Applicants reserve the right to physically incorporate within this description any and all materials and information of such patents or cited publications.

The specific methods and compositions described herein are representative of the preferred embodiments and are exemplary and are not intended as limitations on the scope of the invention. Other objects, aspects and additions will occur to those skilled in the art of the consideration of this description and are encompassed within the spirit of the invention as defined by the attached clauses. It will be readily apparent to the inexpert in the art that various substitutions and modifications can be made to the invention described herein without departing from the scope and spirit of the invention. The illustratively described invention herein suitably may be practiced in the absence of any element or elements or limitation or limitations which are not specifically described as essential. The methods and processes illustratively described herein may be conveniently practiced in different order of steps and these are not necessarily restricted to the step orders indicated herein or in the clauses. As used herein, and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to an "antibody" includes a plurality 'for example, an antibody solution or a series of antibody preparations) of such antibodies and others. Under no circumstances can the patent be interpreted to be limited to the specific examples or incorporations or to the methods especially described herein. Under no circumstances may the patent be construed as being limited by any statement made by any examiner or any other official or employee of the patent and trademark office unless that statement is specifically without qualification or reservation expressly adopted in a written response by the appliers.

The terms and expressions that have been used herein are used as terms of description and not limitation, and therefore there is no intent in the use of such terms and expressions to exclude any equivalent of the features shown and described or parts thereof. , but it is recognized that several modifications are possible within the scope of the invention as claimed. Therefore, it will be understood that while the present invention has been specifically described by preferred embodiments and optional features, modifications and variations of the concepts given herein may be sought by those skilled in the art, and that such modifications and variations are considers that they are within the scope of this invention as defined by the appended claims.

The invention has been described broadly and generally here. Each of the narrower species and sub-generic groupings fall within the generic description that also forms part of the invention. This includes the generic description of the invention with the negative condition or limitation of removing any subject matter of the genre, regardless of or not the cut materials that are specifically recited herein.

Claims (20)

RE I V I N D I C A C I O N S

1. A vaginal insert for increasing vaginal cell growth in a mammal comprising an effective amount of mannose phosphate or a salt thereof, and a carrier, wherein the effective amount is from about 0.001 μg to about 500 mg of phosphate of crafty or a salt thereof.

2. The vaginal insert as claimed in clause 1, characterized in that the insert can increase the production of glycogen by the vaginal epithelial cells.

3. The vaginal insert as claimed in clause 1, characterized in that the insert is used to treat or prevent the proliferation of low vaginal cell, low vaginal cell differentiation or low vaginal moisture.

4. The vaginal insert as claimed in clause 1, characterized in that the insert is used to treat or prevent vaginal atrophy.

5. The vaginal insert as claimed in clause 1, characterized in that the mannose phosphate is a mannose-6-phosphate.

6. The vaginal insert as claimed in clause 1, characterized in that the mannose phosphate is a compound of the formula: wherein X is a monovalent or divalent cation.

7. The vaginal insert as claimed in clause 1, characterized in that the mannose phosphate is mixed with or covalently bound to a polymer.

8. The vaginal insert as claimed in clause 7, characterized in that the polymer is poly (ethylene glycol), poly (vinyl alcohol), poly (vinylpyrrolidone), or poly (2-hydroxyethyl methacrylate).

9. The vaginal insert as claimed in clause 1, characterized in that the composition further comprises an anti-bacterial, anti-fungal or antiviral agent.

10. The vaginal insert as claimed in clause 1, characterized in that the composition further comprises hyaluronic acid.

11. A method for preventing or treating the vaginal condition of a mammal comprising administering intravaginally to a mammal an effective amount of a composition comprising mannose phosphate or a salt thereof wherein the mannose phosphate or the salt thereof can increase the growth of mammalian epithelial cells.

12. The method as claimed in clause 11, characterized in that the condition comprises a vaginal cell proliferation, a low vaginal cell differentiation, a low vaginal moisture or a vaginal atrophy.

13. The method as claimed in clause 11, characterized in that the composition can increase the production of glycogen by vaginal epithelial cells.

14. The method as claimed in clause 11, characterized in that the mannose phosphate is mannose-6-phosphate.

15. The method as claimed in clause 11, characterized in that the mannose phosphate is a compound of the formula: wherein X is a monovalent or divalent cation.

16. The method as claimed in clause 11, characterized in that the mannose phosphate is mixed with or covalently bonded to a polymer.

17. The method as claimed in clause 11, characterized in that the effective amount comprises about 0.001 micrograms to about 500 milligrams of mannose phosphate or salt thereof.

18. The method as claimed in clause 11, characterized in that the composition further comprises an anti-bacterial, anti-fungal or antiviral agent.

19. The method as claimed in clause 11, characterized in that the composition further comprises hyaluronic acid.

20. The use of a composition comprising mannitol phosphate to manufacture a medicament formulated to prevent or treat a vaginal condition comprising low vaginal cell proliferation, low vaginal cell differentiation, vaginal atrophy or low vaginal moisture. SUMMARY The invention provides mannose phosphate and salts thereof to increase vaginal cell growth, vaginal cell maturation and vaginal moisture, as well as compositions, articles and methods for treating and preventing vaginal conditions characterized by vaginal cell growth. poor, a low vaginal cell differentiation and a low vaginal moisture.