High dosage mucoadhesive metronidazole aqueous-based gel formulations their use to treat bacterial vaginosis

Cross-reference of related applications This application claims the interests of US Provisional Patent Application No. 61 / 502,285, filed June 28, 2011, and US Provisional Patent Application No. 61 / 508,058, filed July 14, 2011. Their contents are incorporated herein by reference in their entirety. This application also relates to US Provisional Patent Application No. 61 / 502,288 filed June 28, 2011, and US Provisional Patent Application No. 61 / 508,054 filed July 14, 2011. The contents are incorporated herein by reference in their entirety.

The present disclosure relates to aqueous gel formulations of metronidazole and, for example, their use for treating bacterial vaginosis.

Bacterial vaginosis ("BV") is the most common cause of vaginitis found in women of childbearing age and has been reported to result in 40-50% of all vaginal infections (non-patented). Reference 1). BV can lead to annoying symptoms and can sometimes increase the risk of acquired infections such as HIV. BVs are Gardnerella vaginalis, Bacteroides spp (some now classified as Prevotella spp), anaerobic gram-positive spheres, Mobiluncus spp and mycoplasma ho. Minis (Mycoplasma) It is thought to represent a synergistic multibacterial bacterial infection characterized by overgrowth (100 × ~ 1000 ×) of bacterial species that are often found as part of the normal vaginal microflora, including hominis). With this increase, the number of normally present lactic acid bacteria species (Lactobacillus), which are thought to regulate the growth of other vaginal flora, is significantly reduced. Furthermore, hydrogen peroxide-producing bacteria of lactic acid bacteria, which make up the majority of the vaginal flora of healthy women, are replaced by non-hydrogen peroxide-producing bacteria in women with BV (Non-Patent Document 2). In general, the vaginal microflora of women with BV has more quantitative than qualitative differences compared to healthy women, with one of the clinical signs and symptoms of BV. Studies have shown that it is possible to relate to the above quantitative differences in naturally occurring microbial species (Non-Patent Document 3). Efforts to develop appropriate treatments because the factors involved in the initial disruption of species balance and gradual changes in the vaginal ecosystem are not well understood and the exact location of infection is unknown. Make it difficult.

Metronidazole, and clindamycin, approved by the FDA on July 18, 1963, are the oldest and most commonly used, prescribed to treat women with and / or diagnosed with BV. Two of the antibiotics used ((CDC 2006 STD Treatment Guidelines MMWR 2006; 55 (No.RR-7)).

Metronidazole is available from numerous sources as oral tablets and capsules, injections, 0.75 wt% topical lotions, creams, and gels, 1.0 wt% topical gels, and 0.75 wt% intravaginal gels. Topical creams, lotions, and gels are generally indicated for the treatment of alcohol, and vaginal gels such as 0.75 wt% metronidazole vaginal gel sold by Medicis under the trade name METROGEL VAGINAL® are BVs. Is indicated for treatment of.

Regardless of their general use, treatment with 0.75 wt% intravaginal metronidazole gels such as METROGEL VAGINAL® is not ideal. To be effective, the gel must be applied once or twice daily for a period of 5 days.

In addition, recurrence of BV is commonly observed in approximately 30% of women within 3 months of treatment, whether oral or vaginal. The reason for the recurrence remains unknown. For example, refer to Non-Patent Document 4 and Non-Patent Document 5. A double-blind, placebo-controlled crossmatch showed that intravaginal treatment with 0.75% metronidazole gel resulted in a recurrence rate of approximately 15% one month after treatment. Please refer to Non-Patent Document 6. Vaginal candidiasis, commonly known as yeast infection, is also observed in about 10% of women after treatment for BV. Given the fact that BV is currently the most prevalent form of vaginal infection in women of childbearing age, new treatments that solve one or more of the disadvantages of currently available BV treatments are real. It is necessary immediately and objectively. For example, it provides an effective treatment with a single application and / or provides a more effective treatment than the currently available 0.75 wt% vaginal metronidazole gel and reduces the recurrence rate of BV after a good course of treatment. It is desirable to have available intravaginal treatment that reduces the development of vulvar vaginal candidiasis after a good BV treatment course.

<p num="0008"><nplcit num="1"><text>Sobel, 1997, "Review Article: Vaginitis," New Engl J Med 337: 1896-1903</text></nplcit><nplcit num="2"><text>Amsel et al., 1983, Am J. Med 74:14, Sobel et al, 1990, Infect Med May: 24</text></nplcit><nplcit num="3"><text>Masfari et al., 1986, Genitourin Med 62: 256</text></nplcit><nplcit num="4"><text>Larsson, 1992, Int J Std Aids 3: 239-247</text></nplcit><nplcit num="5"><text>Wilson, 2004, Sex Transm Infect 80: 8-11</text></nplcit><nplcit num="6"><text>Hillier et al., Jun 1993, "Efficacy of Intravaginal 0.75% Metronidazole Gel for the Treatment of Bacterial Vaginosis," Obstet Gynecol 81 (6): 963-967</text></nplcit></p>

<p num="0009"> Formulating metronidazole ("MTZ") in a mucosal adhesive aqueous gel medium at a higher concentration than currently used to treat bacterial vaginosis ("BV") is surprisingly METROGEL VAGINAL (registered). It has been found to provide treatments that overcome some of the shortcomings of current vaginal BV treatments such as (trademark). For example, a "high dose" mucosal adhesive MTZ aqueous gel containing about 1-2% by weight ("% by weight") of MTZ, as detailed below, is surprisingly higher than the expected local concentration of MTZ. Remarkably high concentrations and significantly higher local concentrations of MTZ are delivered by 0.75 wt% MTZ gels similar to METROGEL VAGINAL® in in vitro skin permeation experiments performed on human corpse skin. One embodiment of a high-dose mucosal adhesive aqueous gel containing 1.3 wt% MTZ when used to treat women with and / or diagnosing BV is METROGEL. Excellent results compared to VAGINAL®. For example, when applied once daily for 3 days, the usefulness achieved with the 1.3 wt% high-dose MTZ gels of the present disclosure is their standard with the currently available FDA-approved 0.75 wt% MTZ gels. It was almost identical to that achieved in women treated according to a 5-day treatment regimen. Very surprisingly, a single application of the high dose 1.3 wt% MTZ gel of the present disclosure is as effective as the course of a 0.75 wt% MTZ gel applied once daily for a period of 5 days. Was recognized. This finding is important because it provides an effective treatment for BV with much less total MTZ exposure and reduces the risk of harmful and even more dangerous side effects. The present invention provides, for example: (Item 1) A method of treating subjects with and / or diagnosed with BV, with one or more mucosal adhesive gelled polymers and about 1% to about 2% by weight of metronidazole (MTZ). A method comprising the intravaginal application of a mucosal adhesive aqueous gel containing water to the subject, wherein the amount of the mucosal adhesive aqueous gel applied in a single application contains from about 60 mg to about 100 mg of MTZ. .. (Item 2) METROGEL when the level of MTZ from the mucosal adhesive aqueous gel measured in the stratum corneum and in the receptor is normalized for concentration in in vitro Franzsel skin permeation experiments performed on human corpse skin. The method of item 1, wherein each is at least about 25 to 55 times higher and at least about 1 to 20 times lower than the MTZ level measured from VAGINAL®. (Item 3) The mucosal adhesive aqueous gel has viscosities in the range of about 200,000 mPa to about 400,000 mPa measured at 25 ° C using a controlled shear rate method, the Bohlin CVO100 rheometer, and the rheometer settings in Table 11. , The method described in item 1. (Item 4) The method of item 1, wherein the mucosal adhesive aqueous gel has a degree of mucosal adhesion within about ± 10% of that of MG33PB. (Item 5) The method according to item 1, wherein the mucosal adhesive aqueous gel contains about 30% by weight to about 60% by weight of water. (Item 6) The method of item 1, wherein the mucosal adhesive aqueous gel is substantially free of dextrin, cyclodextrin, niacin, and niacinamide, and optionally a surfactant. (Item 7) The method of item 1, wherein the mucosal adhesive aqueous gel is stable at 25 ° C for a period of at least 6 months. (Item 8) The method of item 1, wherein the mucosal adhesive aqueous gel comprises from about 1% to about 3% by weight total mucosal adhesive gelled polymer and from about 40 to 45% by weight of the MTZ solvent system. (Item 9) 8. The method of item 8, wherein the solvent system comprises one or more solvents having a saturated MTZ solubility of at least about 20 mg / g at 25 ° C. (Item 10) The solvent system has one or more solvents having a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and optionally a saturated MTZ solubility in the range of about 20 mg / g to about 25 mg / g at 25 ° C. 9. The method of item 9, which comprises one or more solvents. (Item 11) The solvent system is one or more lower aromatic alcohols, and optionally one or more lower aliphatic diols and / or one or more polyoxyalkylenes having a molecular weight in the range of about 200 to about 400 ("lower poly"). The method of item 10, comprising "oxyalkylene"). (Item 12) The method of item 11, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and / or one or more lower polyoxyalkylenes. (Item 13) The method of item 12, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more lower polyoxyalkylenes. (Item 14) The method of item 11, wherein the one or more lower aromatic alcohols are selected from the group consisting of benzyl alcohol, 2-methylbenzyl alcohol, phenoxyethanol, and mixtures thereof. (Item 15) Item 10. The item 11 wherein the one or more lower alkylene diols are selected from the group consisting of ethane-1,2-diol (ethylene glycol), propane-1,2-diol (propylene glycol), and mixtures thereof. the method of. (Item 16) The method of item 11, wherein the one or more lower polyoxyalkylenes are selected from the group consisting of polyoxyethylene (polyethylene glycol), polyoxypropylene (polypropylene glycol), and mixtures thereof. (Item 17) The solvent system in which one or more lower aliphatic diols and the one or more lower polyoxyalkylenes have a total lower aliphatic diol: total lower polyoxyalkylene weight ratio in the range of about 1: 1 to about 1: 2. The method according to item 11, which is included in. (Item 18) The solvent system is from about 3.5% to about 5% by weight of the total lower aromatic alcohol, and from about 95% to about 95.5% by weight of the one or more lower alkylene diols and the one or more lower polyoxyalkylenes. 11. The method of item 11, comprising a mixture with. (Item 19) The method of item 18, wherein the weight ratio of total lower alkylene diol: total lower polyoxyalkylene is in the range of about 1: 1 to about 1: 1.67. (Item 20) 19. The method of item 19, wherein the solvent system comprises benzyl alcohol, propane-1,2-diol, and PEG 400. (Item 21) The solvent system comprises from about 3.5% to about 5% by weight of benzyl alcohol and from about 95% to about 96.5% by weight of a mixture of propane-1,2-diol and PEG 400. The method of item 11, wherein the weight ratio of 2-diol: PEG 400 is in the range of about 1: 1 to about 1: 1.67. (Item 22) The method of item 1, wherein the mucosal adhesive aqueous gel further comprises one or more preservatives. (Item 23) The method of item 1, wherein the one or more preservatives are esters of 4-hydroxybenzoic acid. (Item 24) 23. The method of item 23, wherein the one or more preservatives are selected from the group consisting of methyl 4-hydroxybenzoate, propyl 4-hydroxybenzoate, and mixtures thereof. (Item 25) The method according to item 1, wherein the mucosal adhesive aqueous gel has a pH in the range of about pH 3.0 to about pH 5.0. (Item 26) 25. The method of item 25, wherein the mucosal adhesive aqueous gel has a pH of about pH 4.0. (Item 27) The method of item 1, wherein the mucosal adhesive gelled polymer is selected from the group consisting of hydroxyethyl cellulose, carbomer, polycarbophil, and mixtures thereof. (Item 28) The method of item 1, wherein the amount of mucosal adhesive aqueous gel applied in a single application contains approximately 65 mg of MTZ. (Item 29) The method of item 1, wherein the mucosal adhesive aqueous gel comprises from about 1% to about 1.5% by weight of MTZ. (Item 30) 29. The method of item 29, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 1-5 days. (Item 31) 29. The method of item 29, wherein the mucosal adhesive aqueous gel is applied once daily for a period of one day. (Item 32) 29. The method of item 29, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days. (Item 33) The method of item 1, wherein the mucosal adhesive aqueous gel comprises about 1.3% by weight of MTZ. (Item 34) 33. The method of item 33, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 1-5 days. (Item 35) 33. The method of item 33, wherein the mucosal adhesive aqueous gel is applied once daily for a period of one day. (Item 36) 33. The method of item 33, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days. (Item 37) The mucosal adhesive aqueous gel is about 1.3% by weight MTZ, about 2% by weight polycarbofil AA-1, about 2% by weight benzyl alcohol, about 15% by weight propane-1,2-diol, about 25. The method of item 1, comprising weight% PEG400, about 0.02% by weight methyl 4-hydroxybenzoate, and about 0.08% by weight propyl4-hydroxybenzoate. (Item 38) 37. The method of item 37, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 1-5 days. (Item 39) 37. The method of item 37, wherein the mucosal adhesive aqueous gel is applied once daily for a period of one day. (Item 40) 37. The method of item 37, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days. (Item 41) 37. The method of item 37, wherein the amount of mucosal adhesive aqueous gel applied in a single application contains approximately 65 mg of MTZ. (Item 42) A method of treating a subject suffering from and / or being diagnosed with BV, applying a certain amount of aqueous gel intravaginally to the subject a sufficient number of applications to achieve clinical cure. The aqueous gel contains one or more mucosal adhesive polymers, about 1.3% by weight MTZ, and water. (a) In an in vitro Franzcel skin permeation experiment performed on human corpse skin, the level of MTZ from the mucosal adhesive aqueous gel measured in the stratum corneum and in the receptor is normalized for concentration. When METROGEL At least about 25-55 times higher and at least about 1-20 times lower than the MTZ levels measured from VAGINAL®, respectively. (b) Viscosities in the range of about 200,000 mPa to about 400,000 mPa measured at 25 ° C using the controlled shear rate method, the Bohlin CVO100 rheometer, and the rheometer settings in Table 11. (c) Degree of mucosal adhesion within about ± 10% of that of MG33PB, (d) Containing about 30% by weight to about 60% by weight of water, (e) Substantially free of dextrins, cyclodextrins, niacins, and niacin amides, and optionally surfactants, (f) Stable at 25 ° C for at least 6 months, (g) Containing from about 1% to about 3% by weight of total mucosal adhesive polymer and from about 40% to 45% by weight of the solvent system of the MTZ. A method having one or more features or properties selected from the group of. (Item 43) 42. The method of item 42, wherein an amount of the aqueous gel is applied a sufficient number of applications to achieve therapeutic healing. (Item 44) About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of the solvent system of the MTZ, and A mucosal adhesive aqueous gel containing water, MTZ from the mucosal adhesive aqueous gel measured in the stratum corneum and in the receptor in an in vitro Franzcel skin permeation experiment performed on human corpse skin. Mucosal adhesive aqueous gels that, when normalized for concentration, are at least about 25-55 times higher and at least about 1-20 times lower than the MTZ levels measured from METROGEL VAGINAL®, respectively. .. (Item 45) About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of the solvent system of the MTZ, and Mucosa containing water and having viscosities ranging from about 200,000 mPa to about 400,000 mPa measured at 25 ° C using the controlled shear rate method, the Bohlin CVO100 rheometer, and the rheometer settings in Table 11. Adhesive water-based gel. (Item 46) About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of the solvent system of the MTZ, and A mucoadhesive aqueous gel containing water and having a degree of mucosal adhesion within about ± 10% of that of MG33PB. (Item 47) About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of the solvent system of the MTZ, and A mucosal adhesive aqueous gel containing 30% to about 60% by weight of water. (Item 48) About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of the solvent system of the MTZ, and A mucoadhesive aqueous gel containing water and substantially free of dextrin, cyclodextrin, niacin, and niacinamide. (Item 49) About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of the solvent system of the MTZ, and A mucoadhesive aqueous gel that contains water and is stable at about 25 ° C for at least 6 months. (Item 50) 44. The gel of item 44, wherein the mucosal adhesive aqueous gel comprises from about 1% to about 3% by weight total mucosal adhesive gelled polymer and from about 40% to 45% by weight of the MTZ solvent system. .. (Item 51) The gel according to item 50, wherein the solvent system comprises one or more solvents having a saturated MTZ solubility of at least about 20 mg / g at 25 ° C. (Item 52) The solvent system is one or more solvents having a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and optionally a saturated MTZ solubility in the range of about 20 mg / g to about 25 mg / g at 25 ° C. 51. The gel according to item 51, which comprises one or more solvents having. (Item 53) The solvent system is one or more lower aromatic alcohols, and optionally one or more lower aliphatic diols, and / or one or more polyoxyalkylenes having a molecular weight in the range of about 200 to about 400 ("lower"). 52. The gel of item 52, comprising "polyoxyalkylene"). (Item 54) 53. The gel of item 53, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and / or one or more lower polyoxyalkylenes. (Item 55) 54. The gel of item 54, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more lower polyoxyalkylenes. (Item 56) 53. The gel of item 53, wherein the one or more lower aromatic alcohols are selected from the group consisting of benzyl alcohol, 2-methylbenzyl alcohol, phenoxyethanol, and mixtures thereof. (Item 57) 53. Item 53, wherein the one or more lower alkylene diols are selected from the group consisting of ethane-1,2-diol (ethylene glycol), propane-1,2-diol (propylene glycol), and mixtures thereof. Gel. (Item 58) 53. The gel of item 53, wherein the one or more lower polyoxyalkylenes are selected from the group consisting of polyoxyethylene (polyethylene glycol), polyoxypropylene (polypropylene glycol), and mixtures thereof. (Item 59) One or more lower aliphatic diols and the one or more lower polyoxyalkylenes are included in the solvent system in a total lower aliphatic diol: total polyoxyalkylene weight ratio in the range of about 1: 1 to about 1: 2. The gel according to item 53. (Item 60) The solvent system is from about 3.5% to about 5% by weight of the total lower aromatic alcohol, and from about 95% to about 95.5% by weight of the one or more lower alkylene diols and the one or more lower polyoxyalkylenes. 53. The gel according to item 53, which comprises a mixture with. (Item 61) The gel according to item 60, wherein the weight ratio of total lower alkylene diol: total lower polyoxyalkylene is about 1: 1.67. (Item 62) 53. The gel of item 53, wherein the solvent system comprises benzyl alcohol, propane-1,2-diol, and PEG 400. (Item 63) The solvent system comprises from about 3.5% to about 5% by weight of benzyl alcohol and from about 95% to about 96.5% by weight of a mixture of propane-1,2-diol and PEG 400. 2-diol: PEG 53. The gel of item 53, wherein said weight ratio of 400 is in the range of about 1: 1.67 to about 1: 1. (Item 64) 44. The gel of item 44, wherein the mucosal adhesive aqueous gel further comprises one or more preservatives. (Item 65) 64. The gel of item 64, wherein the one or more preservatives are esters of 4-hydroxybenzoic acid (paraben). (Item 66) 65. The gel of item 65, wherein the one or more preservatives are selected from the group consisting of methyl 4-hydroxybenzoate (methylparaben), propyl4-hydroxybenzoate (propylparaben), and mixtures thereof. (Item 67) The gel according to item 44, wherein the mucosal adhesive aqueous gel has a pH in the range of about pH 3.0 to about pH 5.0. (Item 68) 67. The gel of item 67, wherein the mucosal adhesive aqueous gel has a pH of about pH 4.0. (Item 69) 44. The gel of item 44, wherein the mucosal adhesive gelled polymer is selected from the group consisting of hydroxyethyl cellulose, carbomer, polycarbophil, and mixtures thereof. (Item 70) The mucosal adhesive aqueous gel is about 1.3% by weight MTZ, about 2% by weight polycarbofil AA-1, about 2% by weight benzyl alcohol, about 15% by weight propane-1,2-diol, about 25. 44. The gel of item 44, comprising weight% PEG400, about 0.02% by weight methyl 4-hydroxybenzoate, and about 0.08% by weight propyl4-hydroxybenzoate. (Item 71) Mucoadhesive, including about 1.3% by weight MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, and about 40% to about 45% by weight of the MTZ solvent system. Aqueous gel, wherein the solvent system contains benzyl alcohol, propane-1,2-diol, and PEG400, and the mucosal adhesive aqueous gel is at a temperature in the range of about 25 ° C to about 40 ° C. A mucosal adhesive aqueous gel that is stable for a period of 18 months when stored. (Item 72) The gel according to item 71, wherein the one or more mucosal adhesive gelled polymers are selected from the group consisting of crosslinked acrylic acid polymers and cellulose polymers. (Item 73) 72. The gel of item 72, wherein the crosslinked acrylic acid polymer is selected from the group consisting of carbomer and polycarbophil. (Item 74) The gel according to item 72, wherein the cellulose polymer is selected from the group consisting of hydroxyethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose. (Item 75) The gel according to item 71, which comprises about 2% by weight of total mucosal adhesive gelled polymer. (Item 76) A gel according to item 45, which is a unit dose form of a high dose mucosal adhesive MTZ aqueous gel suitable for vaginal application and contains approximately 65 mg to approximately 100 mg of MTZ filled in a container suitable for vaginal application. A unit dose form, including an amount. (Item 77) The unit dose form of item 76, comprising an amount of the gel, comprising about 65 mg of MTZ.</p><p num="0010"> Without being bound by any theory of operation, the surprising usefulness of the illustrated 1.3 wt% MTZ aqueous gel is unexpectedly released as described above by the novel mucosal adhesive aqueous gels described herein. It is thought that this may be due in part to the high concentration of MTZ.</p><p num="0011"> Surprisingly, when a 1.3 wt% MTZ high-dose gel is applied to the vagina of women with BV and / or diagnosed it once daily for a period of 5 days, for example, of cure rate. Compared to METROGEL VAGINAL®, as determined by any one of increased, increased time to resolution of symptoms, increased time to recurrence of symptoms, and / or decreased recurrence rate. It has also been found that its usefulness increases dramatically. Therefore, surprisingly, high-dose mucosal adhesive MTZ aqueous gels are currently used when applied to the vagina of women with BV and / or diagnosed with it once daily for a period of 5 days. It was found that the usefulness achieved was much better and the BV recurrence rate was significantly reduced compared to women treated according to a standard 5-day treatment regimen with a possible FDA-approved 0.75% metronidazole gel. It was.</p><p num="0012"> In addition, women with and / or diagnosed with BV treated for 5 days with the high-dose 1.3 wt% MTZ formulation described herein have other unexpected and prominent results and are treated. There was virtually no subsequent onset of vulvar vaginal candidiasis. As far as the applicant knows, this result is unprecedented.</p><p num="0013"> Thus, in one aspect, the disclosure provides a novel "high dose" mucosal adhesive MTZ aqueous gel that can be applied topically, especially intravaginally, to women, especially to treat BV. High-dose mucosal-adhesive MTZ aqueous gels generally have about 1% to about 2% by weight of MTZ, and in some specific embodiments, about 1.0% by weight, about 1.1% by weight, about 1.2% by weight, about 1.3% by weight. %, About 1.4% by weight, about 1.5% by weight, about 1.6% by weight, about 1.7% by weight, about 1.8% by weight, about 1.9% by weight, or about 2.0% by weight of MTZ, and one or more gelled polymers and / Or include drugs. MTZ may be included in the gel in the form of a free base or salt, eg, a salt formed of a pharmaceutically acceptable acid.</p><p num="0014"> This gel typically uses a controlled shear rate ramp method, a Bohlin CVO100 rheometer, and the rheometer settings in Table 11 below, at about 200,000 to about 400,000 mPa at 25 ° C, eg, at 25 ° C. Contains the total amount of gelled polymer (s) sufficient to produce viscosities in the range of about 250,000 to about 350,000 mPa. Depending on the particular gelled polymer (s) used and the gelling state, aqueous gels with viscosities within this range generally have gelled polymers (s) in the range of about 0.5% to about 5% by weight. Yes) is achieved by including the total amount in the gel. In some specific embodiments, the high dose mucosal adhesive MTZ aqueous gels described herein are from about 1% to about 3% by weight of total gelled polymer (s), and some specifics. In the embodiment, it comprises about 2% by weight of total gelled polymer (s).</p><p num="0015"> Experts will understand that different gelled polymers exhibit different degrees of mucosal adhesion. For gels designed for vaginal application to treat BV, the gelled polymer can be mucosally adhesive. The particular mucosa-adhesive gelled polymer (s) selected is not critical to success and can be selected from any mucosa-adhesive polymer capable of forming a gel in an aqueous solution. An exemplary suitable mucosal adhesive gelled polymer will be further detailed in the detailed description section. All of these mucosal adhesive gelled polymers can be used alone or in combination.</p><p num="0016"> Generally, mucosal adhesive gelled polymers are used to produce aqueous gels with viscosities within the above ranges, using the exemplified high dose mucosal adhesive MTZ aqueous gel MG33PB under the same assay conditions. It should be selected to produce a gel that exhibits a degree of mucosal adhesion within about ± 10% of what is observed. In certain exemplary embodiments, the mucosal adhesive polymer (s) is selected from the group consisting of hydroxyalkyl cellulose, carbomer, polycarbophil, and mixtures thereof. In another particular exemplary embodiment, the mucosal adhesive polymer (s) are polycarbophils.</p><p num="0017"> Experts will understand that in many embodiments, one or more gelled polymers have mucosal adhesive properties, but not necessarily so. Polymers that do not have mucosal adhesion properties or are capable of forming gels in aqueous solutions that produce inadequate degree of mucosal adhesion, with or without gel formation, can be used in combination with mucosal adhesives. The high dose mucosal adhesive MTZ aqueous gels described herein can be produced.</p><p num="0018"> High-dose mucosal adhesive MTZ aqueous gels also include MTZ solvent systems. MTZ is well known to present solubility issues when attempting to formulate gels useful for topical and intravaginal administration with FDA-approved excipients for use in humans. In some cases, MTZ has been attempted to be dissolved in an aqueous solution at a concentration greater than 0.75% by weight or 1.0% by weight. For example, US Pat. No. 6,881,726 describes the use of cyclodextrin and β-cyclodextrin to enhance the solubility of MTZ in aqueous solutions. U.S. Pat. No. 7,348,317 describes the additional use of niacin and niacinamide to enhance the solubility of MTZ in aqueous solutions. In yet another example, surfactants were used to attempt to increase the concentration of MTZ in aqueous solutions. Any of these solvent systems can be used to dissolve the MTZ to the desired concentration of the high dose mucosal adhesive MTZ aqueous gels described herein.</p><p num="0019"> Dissolution accelerators can be used, but surprisingly, MTZ is a high-dose mucosal adhesive MTZ aqueous system containing about 1% to about 2% by weight MTZ without the need to use lysis-promoting compounds such as those mentioned above. It has been discovered that it can be dissolved in sufficient concentrations to produce a gel. Cyclodextrin and β-cyclodextrin often enhance the water solubility of the compound by forming a complex, resulting in a complex formulation in which the compound is not uniformly dissolved in the formulation, and drugs such as niacin and nycinamide This is advantageous because it itself has undesired pharmacological properties at certain concentrations and agents such as surfactants can polyphase the gel.</p><p num="0020"> Therefore, novel solvent systems are also provided that are useful for preparing the high dose MTZ gels described herein. The novel solvent system yields a homogeneous, high-dose mucosal adhesive MTZ aqueous gel, i.e., when the MTZ is dissolved in the gel and stored at a temperature in the range of about 25 ° C to about 40 ° C, for a long period of time For example, it can be stored for 6 months or more. In fact, certain exemplary embodiments of high-dose mucosal adhesive MTZ aqueous gels containing 1.3 wt% MTZ are stable for at least 18 months when stored at temperatures from about 25 ° C to about 40 ° C. It turned out that there was.</p><p num="0021"> The novel solvent system generally utilizes one or more solvents that collectively have sufficient saturated MTZ solubility at 25 ° C to produce a gel containing the desired concentration of MTZ. Generally, such solvents each have a saturated MTZ solubility of at least about 10 mg / g at 25 ° C, but if a skilled person chooses a solvent with a clearly high saturated MTZ solubility, the new solvent system It will be appreciated that it may contain solvents with lower saturated MTZ solubility. A non-limiting list of solvents with suitable saturated MTZ solvents that can be used as components of the novel solvent system is provided in the Examples section.</p><p num="0022"> A class of solvents was found to have a saturated MTZ solubility of at least about 20 mg / g at 25 ° C. For example, there are certain alcohols, such as lower aliphatic alcohols such as ethanol and lower aromatic alcohols such as benzyl alcohol, such as lower aliphatic diols (including, for example, lower aliphatic glycols such as ethylene glycol and propylene glycol). Diols, as well as polyethers such as, for example, polyoxyalkylenes ("lower polyoxyalkylenes") having molecular weights in the range of about 200 to about 600 (including, for example, PEG400), are saturated MTZs in this range at 25 ° C. It was found to have solubility. In fact, some lower aromatic alcohols have a significantly higher saturated MTZ solubility at 25 ° C, for example at least about 50 mg / g. As a particular example, benzyl alcohol has a saturated MTZ solubility of about 72 mg / g at 25 ° C. All of these solvents, and others with a saturated MTZ solubility of at least about 20 mg / g at 25 ° C, are described herein in a novel solvent system alone and in various combinations. MTZ can be dissolved in high-dose mucosal adhesive MTZ aqueous gels.</p><p num="0023"> In some embodiments, the novel solvent system has at least one solvent having a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and optionally at least about 20 mg / g of saturated MTZ solubility at 25 ° C. Contains one or more solvents that have. In some embodiments, the novel solvent system aggregates at least one solvent having a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and a saturated MTZ solubility of at least about 20 mg / g at 25 ° C. Contains one or more solvents that have in. In some particular embodiments, each of the solvents has a saturated MTZ solubility of at least about 20 mg / g at 25 ° C.</p><p num="0024"> In some particular embodiments, the novel solvent system comprises at least one solvent from about 1% to about 5% by weight, having a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and the rest. About 95% to about 99% by weight is composed of one or more solvents that collectively have a saturated MTZ solubility of at least about 20 mg / mL at 25 ° C. In some specific embodiments, the solvent system is about 1.0% by weight, about 1.5% by weight, about 2.0% by weight, about 2.5% by weight, about 3.0% by weight, about 3.5% by weight, about 4.0% by weight, about 4.5% by weight. Contains at least one solvent with a saturated MTZ solubility of at least about 50 mg / mL at 25 ° C by weight%, or about 5.0% by weight (or any range enclosed by any of these values), the rest 25 ° C. Consists of one or more solvents that collectively have a saturated MTZ solubility of at least about 20 mg / mL in C.</p><p num="0025"> It has been found that ternary solvent systems utilizing one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more lower polyoxyalkylenes provide good results. Therefore, in some specific embodiments, the high dose mucosal adhesive MTZ aqueous gels described herein are one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more. Utilizes a novel solvent system of MTZ, including the lower polyoxyalkylenes of.</p><p num="0026"> In some specific embodiments, the solvent systems described herein enhance the solubility of MTZ in aqueous solutions such as cyclodextrin, β-cyclodextrin, niacin, niacinamide, and / or surfactants. Contains or substantially does not contain other drugs known to do. In some specific embodiments, the solvent system comprises only one or more lower aromatic alcohols, one or more lower alkylenediols, and one or more polyoxyalkylenes and any other additional agent or It is a ternary solvent system that does not contain a solvent.</p><p num="0027"> The one or more lower aromatic alcohols can be phenolic, primary, secondary, or tertiary alcohols and thus may include aliphatic or heteroaliphatic groups in addition to aromatic groups. The one or more lower aliphatic diols and lower polyoxyalkylenes can be saturated or unsaturated and may contain primary, secondary, and / or tertiary alcohol groups. Illustrative lower aromatic alcohols, lower aliphatic diols, and lower polyoxyalkylenes useful in this particular embodiment of the novel solvent-based and high-dose mucosal adhesive MTZ aqueous gels described herein are detailed. More details in the explanation section. In some particular embodiments, the one or more lower aromatic alcohols are benzyl alcohols and the one or more lower aliphatic diols are propane-1,2-diols (propylene glycol), one. The above lower polyoxyalkylene is a polyethylene glycol having about 400 MW (PEG400).</p><p num="0028"> In some specific embodiments, the solvent system is from about 3.5% to about 5% by weight of the total lower aromatic alcohol (s), and from about 95% to about 96.5% by weight of the total lower aliphatic diol (s). Approximately 95% to 96.5% by weight of total lower polyoxyalkylenes (s), or approximately 95% to 96.5% by weight of total lower aliphatic diols (s) and total lower polyoxyalkylenes (s) Includes a mixture of (s). When the mixture is used, the weight ratio of total lower aliphatic diol (s): total lower polyoxyalkylene (s) may typically range from about 1: 1 to about 1: 2. , In some specific embodiments it is about 1: 1.67.</p><p num="0029"> Solvent systems generally consist of high dose mucosal adhesive MTZ aqueous gels ranging from about 30% to about 60% by weight. The exact amount used depends at least in part on the desired amount of MTZ contained in the gel. In some specific embodiments, the solvent system represents a high dose mucosal adhesive MTZ aqueous gel of about 40-45% by weight.</p><p num="0030"> The amount of solvent-based components can also be explained for high-dose mucosal-adhesive MTZ aqueous gels. In some specific embodiments, the high dose mucosal adhesive MTZ aqueous gel is from about 1.5% to about 2% by weight of total lower aromatic alcohols (s) (eg, benzyl alcohol), from about 15% by weight. Approximately 25% by weight of total lower aliphatic diols (s) (eg, propane-1,2-diol) and / or approximately 15% to approximately 25% by weight of total lower polyoxyalkylenes (s) (eg, PEG). 400) is included. In some specific embodiments, the high dose mucosal adhesive MTZ aqueous gel is about 1.5% to about 2% by weight of the total lower aromatic alcohol (s) (eg, benzyl alcohol), about 15% by weight. Includes total lower aliphatic diols (s) (eg, propane-1,2-diol) and / or about 25% by weight total lower polyoxyalkylenes (s) (eg, PEG 400). Other specific embodiments are described in the detailed description section as additional solvents and agents useful for dissolving MTZ in combination with the high dose mucosal adhesive MTZ aqueous gels described herein. Will be done.</p><p num="0031"> The pH of the high-dose mucosal adhesive MTZ aqueous gels described herein should generally match the pH of the intended application region, eg, when intended for vaginal application, the pH of a healthy vagina. Should match. Without being bound by any particular theory, matching the pH of a healthy vagina may allow beneficial lactic acid bacteria to grow. Therefore, for gels intended for vaginal application, the pH should generally be in the range of about pH 3 to about pH 5, eg, about pH 4. The pH can be adjusted and / or maintained with acids, bases, and buffers, as is well known in the art. For example, the pH of the gel may be adjusted and / or maintained by utilizing a buffer with appropriate normality and pH as the aqueous phase of the gel. Alternatively, the pH of the aqueous phase may be adjusted with an acid or base prior to adding the gelling agent. After gelation, the pH may be further adjusted with acid or base. This latter method can be advantageous for preparing gels utilizing gelling agents that gel most efficiently outside the desired pH range of the resulting gel. For example, carbomer gel gels most efficiently around neutral pH. A high-dose mucosal adhesive MTZ aqueous gel suitable for vaginal application adjusts the pH of the gelling solution to approximately neutral for gelling, and then uses an acid to bring the resulting gel to a pH of approximately pH 3 ~. It may be obtained by adjusting the pH within the range of about 5. However, high dose mucosal adhesive MTZ aqueous gels utilizing certain pH sensitive mucosal adhesive gelled polymers such as carbomer and polycarbofil polymers, which have a pH within the desired range for intravaginal application, are buffered or It was discovered that it was obtained without pH adjustment.</p><p num="0032"> The high-dose mucosal adhesive MTZ aqueous gels described herein can also contain other additional components, such as one or more preservatives, as is well known in the art. The preservative (s) should generally contain only about 1% or 2% by weight of a high dose mucosal adhesive MTZ aqueous gel when used. The choice of preservative (s) is not important. Suitable useful preservatives are further detailed in the detailed description section. In some particular embodiments, the one or more preservatives (s) are esters of 4-hydroxybenzoic acid, also known as parabens. Suitable parabens include lower alkyl esters of 4-hydroxybenzoic acid, such as methyl 4-hydroxybenzoate (methyl paraben), ethyl 4-hydroxybenzoate (ethyl paraben), and propyl 4-hydroxy-benzoate (propyl paraben).</p><p num="0033"> Experts will understand that the solvents used to dissolve MTZ in the high dose mucosal adhesive MTZ aqueous gels described herein may have storage properties. As a particular example, benzyl alcohol has well-known storage properties. When used as a solvent in a new solvent system, this storage property can be used to benefit. In fact, gels containing solvents with preservative properties do not necessarily have to contain additional preservatives. Gels that utilize solvent-based storage properties decompose so that the gel retains the amount of undecomposed or unoxidized solvent that has effective storage properties after a desired period of time if the solvent can decompose or oxidize over time. And / or contains an excess amount that takes into account oxidation. For example, benzyl alcohol is known to oxidize to benzaldehyde and has no storage properties. In embodiments of high-dose mucoadhesive MTZ aqueous gels described herein using benzyl alcohol as both the MTZ solvent and preservative, an amount of benzyl alcohol should be included in the gel and the MTZ. In addition to dissolving the gel, it provides a shelf life during the expected shelf life of the gel. Excess amounts of benzyl alcohol or other solvents used in solvent systems that are partially used as preservatives can be determined based on the degradation and / or oxidation properties and kinetics of a particular solvent under desired storage conditions. ..</p><p num="0034"> Embodiments of high-dose mucosal adhesive MTZ aqueous gels containing solvents with preservative properties include one or more additional preservatives and / or preservatives designed to protect the solvent from decomposition and / or oxidation. It may be included. For example, in the case of benzyl alcohol, the high dose gels described herein may contain one or more additional preservatives with antioxidant properties that partially protect benzyl alcohol from oxidation. In certain embodiments, high dose mucosal adhesive MTZ aqueous gels containing benzyl alcohol or other solvent that oxidizes include one or more parabens as an additional preservative. In some specific embodiments, the high dose mucosal adhesive MTZ aqueous gel is about 0.1% by weight total paraben, in some specific embodiments about 0.02% by weight methylparaben, and about 0.08% by weight propyl. Including parabens.</p><p num="0035"> High-dose mucosal adhesive MTZ aqueous gels also contain water in either the form of pure water or in the form of an aqueous buffer. Typically, the amount of water contained in the gel is less than about 70% by weight, more typically less than about 60% by weight, and in some particular embodiments, in the range of about 45% to about 55% by weight. Is.</p><p num="0036"> As mentioned above, some embodiments of high-dose mucosal adhesive MTZ aqueous gels prepared in the novel solvent systems described herein use Franz cell with human corpse skin (and silicone membranes). In an in vitro skin permeation experiment performed in, an unexpectedly high local concentration of MTZ and a significantly higher local concentration of MTZ than the conventional 0.75 wt% MTZ gel are delivered. In fact, as further detailed in the Examples section, virtually all tested high-dose MTZ aqueous gels were of the conventional 0.75 wt% MTZ gel when normalized for concentration against the stratum corneum. More MTZ was delivered locally in the range of about 25 to about 55 times. Conventional in vitro skin permeation experiments performed in Franzsel with human corpse skin, regardless of the higher local MTZ concentrations delivered by the high-dose mucosal adhesive MTZ aqueous gels described herein. Significantly less MTZ was observed in the receptor compared to the 0.75 wt% MTZ gel. As further detailed in the detailed description section, virtually all high-dose MTZ gels tested in this experiment, when normalized for concentration, were approximately 1 ~ of the conventional 0.75 wt% MTZ gel. Less MTZ was produced in the receptor in the range of about 20-fold.</p><p num="0037"> Without being bound by any theory, these properties allow high local concentrations of MTZ to be delivered, potentially increasing usefulness, while at the same time reducing systemic exposure in some topical applications. It is considered to be meaningful and important for therapeutic application because it reduces unwanted side effects. Therefore, in some embodiments, the various components of the high-dose mucosal adhesive MTZ aqueous gel are the surface of the MTZ measured in in vitro Franzsel skin permeation experiments performed with human corpse skin or silicone membranes. When the levels are normalized for concentration, they are at least about 25 times higher than the levels measured using conventional 0.75 wt% MTZ gels such as METROGEL VAGINAL®, and in some specific embodiments, about. 55 times higher. In some embodiments, the various components of the high-dose mucosal adhesive MTZ aqueous gel have the levels of MTZ measured in the receptor in an in vitro Franzsel skin permeation experiment performed on human corpse skin. METROGEL, when normalized for concentration To produce high-dose gels that are at least about 1-fold lower than the levels measured using conventional 0.75 wt% MTZ gels such as VAGINAL® and, in some specific embodiments, about 20-fold lower. Be selected. Specific in vitro Franzsel experiments can be used in the comparative studies described in Example 6.</p><p num="0038"> The high-dose mucosal adhesive MTZ aqueous gels described herein can be used for any purpose for which topical treatment of MTZ is desirable. Due to their degree of mucosal adhesion, they are particularly useful for treating women who suffer from BV and / or have been diagnosed with it.</p><p num="0039"> Therefore, in another embodiment, the disclosure describes a method of treating a woman suffering from and / or being diagnosed with BV using the high dose mucosal adhesive MTZ aqueous gel described herein. provide. The method generally uses a sufficient amount of the high-dose mucosal adhesive MTZ aqueous gel described herein to provide a therapeutic effect in the vagina of women suffering from and / or being diagnosed with BV. Accompanied by internal application. The clinical criteria for diagnosis as well as the criteria for establishing therapeutic efficacy are further detailed in the detailed description section. In some specific embodiments, the amount of high dose mucosal adhesive MTZ aqueous gel applied in a single application contains from about 60 mg to about 100 mg of MTZ. In other specific embodiments, the amount of high-dose mucosal adhesive MTZ aqueous gel applied in a single application is from about 60 mg to about 90 mg of MTZ, and in yet other specific embodiments, from about 60 mg to about 80 mg. It contains MTZ, and in yet other particular embodiments, about 60 mg to about 70 mg of MTZ. In yet other specific embodiments, the amount of high-dose mucosal adhesive MTZ aqueous gel applied in a single application contains approximately 65 mg of MTZ.</p><p num="0040"> The frequency and duration of application may vary and may depend on the desired outcome. Generally, the gel is applied once daily for a period of 1, 2, 3, 4, or 5 days. Significant therapeutic effects were found to be achieved with a single application of a high dose mucosal adhesive MTZ aqueous gel containing approximately 65 mg of MTZ. Therefore, in some embodiments, the high dose mucosal adhesive MTZ aqueous gel is applied as a single application. Applying a high-dose mucosal adhesive MTZ aqueous gel containing approximately 65 mg MTZ once daily for a period of 5 days resulted in a lower BV recurrence rate than a similar course of treatment with an FDA-approved 0.75 wt% MTZ gel, in fact. It was also found that vulvar vaginal candidiasis did not occur after treatment. Therefore, in some embodiments, the high dose mucosal adhesive MTZ aqueous gel is applied once daily for a period of 5 days.</p><p num="0041"> For use, the high dose mucosal adhesive MTZ aqueous gel can be encapsulated in any form convenient for the desired mode of application. In certain embodiments useful for vaginal application, the high-dose mucosal adhesive MTZ aqueous gel is encapsulated in a single-dose form, such as a pre-filled single-dose syringe-type applicator, as a particular example. Good.</p><p num="0042"> When used to treat women with and / or diagnosed with BV, the high dose mucosal adhesive MTZ aqueous gels described herein achieve, for example, (1) therapeutic effects. Single, unlike existing treatments with 0.75 wt% MTZ gel, eg METROGEL VAGINAL®, which requires administration once daily or twice daily for a period of 5 days. Effective treatment of BV following application, as well as (2) increased cure rate, decreased time to resolve symptoms, increased time to recurrence of symptoms, and / or incidence of genital vaginal candidiasis It offers a number of surprising benefits, including increased usefulness compared to METROGEL VAGINAL® following once-daily, 5-day application, as determined by any one of the reductions.</p><p num="0043"> It should be understood that the above abstracts do not describe all embodiments or implementations of the various inventions disclosed herein. The detailed description and examples section further exemplifies specific embodiments. The various embodiments described herein are to be disclosed in combination as if each particular combination were explicitly disclosed. The examples are merely representative and should not be construed as exclusive or limiting the scope of the various inventions disclosed herein.</p>

<figref num="1">The graphs showing the mucosal adhesion properties of the exemplary high dose mucosal adhesive MTZ aqueous gels MG32PB and MG33PB to the porcine gastric mucosa are provided. The data show the percentage of MTZ remaining in the gastric mucosa as a function of time. For this experiment, data were taken every 15 seconds for the first 5 minutes.</figref><figref num="2">The graphs showing the mucosal adhesion properties of the exemplary high dose mucosal adhesive MTZ aqueous gels MG32PB and MG33PB to the porcine gastric mucosa are provided. The data show the percentage of MTZ remaining in the gastric mucosa as a function of time. For this experiment, data were taken every minute for the first 15 minutes.</figref><figref num="3A">In vitro Franzsel skin permeation experiments performed on human corpse skin, recovered from various skin layers of various exemplary high-dose mucosal adhesive MTZ aqueous gels.¹⁴A graph showing the average amount (μg; ± standard error) of C-labeled MTZ is provided.</figref><figref num="3B">In vitro Franzsel skin permeation experiments performed on human corpse skin, recovered from various skin layers of various exemplary high-dose mucosal adhesive MTZ aqueous gels.¹⁴A graph showing the average amount (μg; ± standard error) of C-labeled MTZ is provided.</figref><figref num="4A">Total applied¹⁴Graphs showing the recovery data in Figures 3A and 3B are provided, respectively, expressed as the mean percentage (± standard error) of the C-labeled MTZ.</figref><figref num="4B">Total applied¹⁴Graphs showing the recovery data in Figures 3A and 3B are provided, respectively, expressed as the mean percentage (± standard error) of the C-labeled MTZ.</figref><figref num="5A">Graphs are provided showing different views of the recovered data in FIGS. 3A and 3B, respectively.</figref><figref num="5B">Graphs are provided showing different views of the recovered data in FIGS. 3A and 3B, respectively.</figref><figref num="6A">Graphs are provided showing different views of the recovered data in FIGS. 4A and 4B, respectively.</figref><figref num="6B">Graphs are provided showing different views of the recovered data in FIGS. 4A and 4B, respectively.</figref><figref num="7">Average amount of MTZ recovered in surface and vaginal tissue obtained from the exemplary high-dose MTZ mucosal adhesive aqueous gels and control gels 24 hours after application in in vitro Franzsel experiments performed with porcine vaginal tissue (μg) ) Is provided. The data are mean ± standard error (n = 5).</figref><figref num="8">Pig vaginal tissue (μg / cm) in the Franz cell experiment in Figure 7.²A graph showing the average amount of MTZ permeated through vaginal tissue) is provided. The data are mean ± standard error (n = 5).</figref><figref num="9">A graph comparing ATP released from a coating sample infected with Gardnerella placebo following 24-hour treatment with a different exemplary high-dose mucosal-adhesive MTZ aqueous gel is provided (n = 3 for active gels). , Placebo gel and control n = 2).</figref><figref num="10">Provides a graph comparing ATP released from epithelial samples infected with Gardnerella placebo after treatment with the exemplary high-dose MTZ gel MG33PB at different dosing times of 4, 8, and 24 hours (for MG33PB). n = 3, placebo gel and control n = 2).</figref><figref num="11">A graph comparing ATP released from a coating sample infected with Gardnerella placebo following a 2-hour treatment with a different exemplary high-dose mucosal-adhesive MTZ aqueous gel is provided (n = 6 for active gels). , Placebo gel and control n = 3).</figref><figref num="12">Provided are graphs showing regions of inhibition of growth of the illustrated high-dose mucosal adhesive MTZ aqueous gel MG33PB and various placebo versions of Gardonerella baginalis containing different amounts of benzyl alcohol.</figref>

A more complete understanding of the various inventions disclosed herein, and many of the benefits associated therewith, are provided by the detailed description below. 6.1. Definition

As used herein in its entirety and in the appended claims, the following terms and expressions shall have the following meanings:

The indefinite articles "a" and "an", as well as the definite articles "the", shall include both the singular and the plural unless the context in which they are used explicitly states otherwise.

"At least one" and "one or more" are used synonymously to mean that the item may contain one or more of the listed elements.

Unless otherwise indicated, all numerical values used herein and in the claims to describe the amounts, ratios, numerical properties, reaction conditions, etc. of the components are in any case amended by the term "about". Understand what you think you will get. 6.2. Detailed description of a particular exemplary embodiment

As described in the abstract, the present disclosure suffers from bacterial vaginosis ("BV"), or in certain embodiments, particularly useful for topical delivery of MTZ for the treatment of various microbial infections. Specially provided are high-dose mucosal adhesive metronidazole (MTZ) aqueous gels useful for vaginal application as a therapeutic approach to the treatment of women who have been diagnosed with it. High-dose mucosal adhesive MTZ aqueous gels generally include MTZ and one or more gelled polymers, and in certain embodiments, include one or more mucosal adhesive gelled polymers.

MTZ, also known as 2- (2-methyl-5-nitro-1H-imidazol-1-yl) ethanol, is a genus of Fuzobacterium and Bacteroides (eg, B.fragilis, B.distasonis, B.ovatus, B.thetaioaomicron). , And B. vulgates), anaerobic gram-negative rods, including susceptibility strains of Clostridium and eubacteria, and well-known anaerobic gram-positive bacilli, including Peptostreptococcus. It is an antibacterial agent.

MTZ may be included in the high dose gels described herein in the form of free bases or as salts formed with pharmaceutically acceptable acids. Suitable inorganic acids for forming pharmaceutically acceptable salts are, but are not limited to, examples of hydrohalogen acids (eg, hydrochloride, hydrobromic acid, hydroiodic acid, etc.), sulfuric acid, nitric acid. , Phosphoric acid and the like. Suitable organic acids for forming pharmaceutically acceptable salts include, but are not limited to, examples of acetic acid, trifluoroacetic acid, propionic acid, sulphonic acid, cyclopentanepropionic acid, glycolic acid, oxalic acid, pyruvate. , Lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, palmitic acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, transdermal acid, mandelic acid, alkyl sulfonic acid (For example, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, etc.), arylsulfonic acid (eg, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, etc.) , 4-Toluene Sulfonic Acid, Camper Sulfonic Acid, etc.), 4-Methylbicyclo [2.2.2]-oct-2-en-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid , Lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid.

MTZ can be commercially available or synthesized by well-known methods.

The high dose mucosal adhesive MTZ aqueous gels described herein generally contain an amount of MTZ in the range of about 1% to about 2% by weight. In certain embodiments, the gels described herein are about 1.0% by weight, about 1.1% by weight, about 1.2% by weight, about 1.3% by weight, about 1.4% by weight, about 1.5% by weight, about 1.6% by weight. Includes about 1.7% by weight, about 1.8% by weight, about 1.9% by weight, or about 2.0% by weight of MTZ.

High-dose mucoadhesive MTZ aqueous gels are one or more gelled polymers and, in certain embodiments, one or more mucoadhesive gelled polymers that impart their mucosal adhesiveness and gel-like properties to the gel. Also includes. Various polymers that form mucosal adhesive gels in aqueous solutions suitable for use in mucoadhesive aqueous gels described herein are known in the art and are not limited, but by way of example, by way of example. Polysaccharide hydrophilic colloids (including, for example, mucilage, gums such as xanthan gum and tragacant, and glucan), cellulose and modified cellulose (including, for example, alkyl cellulose, hydroxyalkyl cellulose, carboxy cellulose, and sodium carboxy cellulose), polymers (polyoxy). Copolymers of ethylene and polyoxypropylene, also known as PLURONICS®, carbomer (crosslinked polymer of acrylic acid), polycarbofyl (polymer of polyacrylic acid crosslinked with divinyl glycol), veegum (aluminum silicate) Includes (magnesium), polyvinyl alcohol (PVA), gelatin, sodium arginate, and polyvinylpyrrolidone (PVP). An exemplary suitable mucus can be found, for example, in Malviya et al, 2011, "Applications of Mucilages in Drug Delivery-A Review," Advan Biol Res 5 (1): 1-7, and references cited therein. These disclosures can be incorporated herein by reference.

In some specific embodiments, the gelled polymer (s) are crosslinked polymers of acrylic acid, such as, for example, carbomer or polycarbophils, and / or cellulose polymers. Suitable cellulose polymers include, but are not limited to, carboxymethyl cellulose (CMC), methyl cellulose, ethyl cellulose, hydroxyethyl cellulose (NEC or HHX), hydroxylpropyl cellulose (HPC), and hydroxylpropyl methylcellulose (HPMC). Suitable carbomeres include, but are not limited to, various polymers sold by Lubrizol Advanced Materials (Cleveland, Ohio) under the trade name CARBOPOL®, such as CARBOPOL Homopolymers (allyl scroll or allyl) Polymer of acrylic acid cross-linked with pentaerythritol), eg CARBOPOL® 71G NF, CARBOPOL® 971P NF, CARBOPOL® 974P NF, CARBOPOL® 980 NF, and CARBOPOL® 981 NF; CARBOPOL® copolymers (polymers of acrylic acid and C10-C30 alkyl acrylate crosslinked with allylpentaerythritol), such as PEMULEN TR-1NF and PEMULEN . TR-2 NF; CARBOPOL® interpolymers (carbomer homopolymers or copolymers containing block copolymers of polyethylene glycol and long chain alkyl acid esters) such as CARBOPOL® ETD 2020 NF and CARBOPOL® Ultrez 10 NF; "conventional" polymers such as Carbomer 934 (CARBOPOL® 934 NF), Carbomer 934P (CARBOPOL® 934P NF), Carbomer 940 (CARBOPOL® 940 NF), Carbomer 941 (CARBOPOL) (Registered Trademarks) 941 NF), and Carbomer 1342 (CARBOPOL® 1342P NF), and PolyCarbophil (NOVEON® AA-1). USP).

Any of these and / or other mucosal adhesive gelled polymers can be used alone or in combination in the high dose mucosal adhesive MTZ aqueous gels described herein.

One or more mucosal adhesive gelled polymers typically yield high dose mucosal adhesive MTZ aqueous gels in the range of about 200,000 to 400,000 mPa at 25 ° C, in some specific embodiments 25. Used in an amount such that it has a viscosity in the range of about 250,000-350,000 mPa at ° C, this viscosity is controlled by the shear rate ramp method, Bohlin. Measured using the CVO100 rheometer and the rheometer settings listed in Table 11 (hereafter, Example 5). Depending on the particular gelled polymer (s) used and the gelling conditions, aqueous gels with viscosities within this range generally have gelled polymers (s) in the range of about 0.5% to about 5% by weight. ) Is included in the gel. In some specific embodiments, the high dose mucosal adhesive MTZ aqueous gels described herein contain from about 1% to about 3% by weight of total gelling polymer (s) and some. In certain embodiments, it comprises about 2% by weight of total gelled polymer (s).

Gels designated for vaginal application should ideally exhibit a degree of mucosal adhesion that prevents the gel from leaking during application. Gels that utilize cellulose and / or acrylic acid mucosal adhesive gelled polymers with viscosities within the above ranges need to have an appropriate degree of mucosal adhesion. Different gelled polymers exhibit different degrees of mucosal adhesion. Gelled polymers (listed in Examples 2 and 5) that produce high-dose mucosa-adhesive MTZ aqueous gels with a degree of mucosal adhesion within about ± 10% of that of the exemplary gel MG33PB are suitable. Can be used. In certain exemplary embodiments, the mucosal adhesive polymer (s) is selected from the group consisting of hydroxyalkyl cellulose, carbomer, polycarbophil, and mixtures thereof. In another particular exemplary embodiment, the mucosal adhesive polymer (s) is a polycarbophil, eg, a polycarbophil sold by Lubrizol, Inc. under the trade name NOVEON® AA-1 polycarbophil. Is.

A high-dose mucosal adhesive MTZ aqueous gel with suitable viscosity, mucosal adhesion, and other desirable properties for utilizing Carbomer 934, hydroxyethyl cellulose, or polycarbofil as a mucosal adhesive gelled polymer is described in Example 2. Provided.

As described in the abstract, the solubility of MTZ presents a problem when attempting to formulate MTZ in aqueous formulations such as aqueous gels. Certain embodiments of high-dose mucosal adhesive MTZ aqueous gels described herein are long-term, eg, at least about 6 months, when stored at temperatures ranging from about 25 ° C to about 40 ° C. Utilize a novel solvent system that has been found to produce homogeneous gels containing MTZ at high concentrations of stable 2% by weight or higher. Surprisingly, such high dose mucosal adhesive MTZ aqueous gels are commonly used to enhance the solubility of MTZ in aqueous solutions such as cyclodextrin, β-cyclodextrin, niacin, niacinamide, and / or surfactants. It can be prepared without using any chemicals.

As used herein, "stable" means that the gel has the following properties: (1) MTZ purity of at least about 95% when stored at a temperature of about 25 ° C for a period of at least 6 weeks. (2) When stored for a period of at least 6 weeks at a temperature of about 25 ° C, there is a significant increase in crystal and / or particle formation compared to reference values during microscopic visual inspection at 40 × magnification. Not (for example, only about 2 to 3 times increase), (3) When stored at a temperature of about 25 ° C for a period of about 6 weeks, at a reference value or at a temperature within the range of about 2 to 8 ° C Means that it exhibits only about ± 50% change in viscosity as measured at 25 ° C compared to the stored control sample. In addition, the stable gel has the following additional properties: (4) 6 weeks at a temperature of about 25 ° C compared to a reference sample or a control sample stored at a temperature in the range of about 2-8 ° C. When stored for a period of about 25 ° C, compared to pH changes of less than about ± 0.3 pH units, (5) reference values or control samples stored at temperatures in the range of about 2-8 ° C. It is desirable, but not essential, to exhibit a change in the usefulness of the preservative of only about ± 20% when stored at the same temperature for a period of at least 6 weeks. Assays suitable for measuring the stability of high-dose mucosal adhesive MTZ aqueous gels are provided in Examples 4 and 5. Typically, the particular constituents and amounts of the novel solvent system are selected to produce stable, high dose mucosal adhesive MTZ aqueous gels as defined herein. Guidance for selecting useful solvent systems for preparing stable, high-dose MTZ aqueous gels with a particular amount of MTZ is provided by the various exemplary high-dose gels disclosed in the Examples section.

In some specific embodiments, the components and amounts of the novel solvent system, when stored at a temperature of about 40 ° C for a period of about 6 weeks, the MTZ purity, homogeneity, and viscosity properties described above, as well as It is optionally selected to produce a stable, high-dose mucosal-adhesive MTZ aqueous gel with the above-mentioned pH and storage potency properties. In other specific embodiments, the components and amounts of the novel solvent system are at temperatures in the range of about 25-40 ° C, and in certain embodiments at temperatures of about 25 ° C or about 40 ° C for 6 months. Stable, high-dose mucoadhesive MTZ with the above-mentioned MTZ purity, homogeneity, and viscosity properties, and optionally the above-mentioned pH and storage efficacy properties, when stored for a period of time or longer, for example as long as 18 months. Selected to produce aqueous gels. In yet other specific embodiments, the components and amounts of the novel solvent system are at temperatures in the range of about 25-40 ° C, and in certain embodiments at temperatures of about 25 ° C or about 40 ° C. Stable, high-dose mucosal adhesion with the above-mentioned MTZ purity, homogeneity, viscosity, pH, and optionally the above-mentioned storage efficacy properties when stored for a period of 6 months or longer, for example as long as about 18 months. Selected to produce MTZ aqueous gels.

Various new solvent systems are described in the summary section above. In some specific embodiments, the novel solvent system is of one or more lower aromatic alcohols, one or more lower aliphatic diols, and / or about 200 to about 600 ("lower polyoxyalkylenes"). A ternary system containing one or more polyoxyalkylenes having a MW in the range.

As used herein, a "lower alcohol" comprises saturated and unsaturated non-aromatic and aromatic alcohols having 1 to 15 carbon atoms, optionally one or more heteroatoms, eg, It may contain one or more oxygen atoms instead of carbon atoms (s). Lower non-aromatic alcohols may include linear, branched, or cyclic primary, secondary, or tertiary lower aliphatic alcohols and lower heteroaliphatic alcohols. Examples of lower aliphatic alcohols include, but are not limited to, methanol, ethanol, propane-1-ol, propane-2-ol, butane-1-ol, butane-2-ol, 2-methylpropane-1-. Ol, 2-methylpropane-2-ol, pentane-1-ol, pental-2-ol, pentane-3-ol, 3-methylbutane-1-ol, hexane-1-ol, hexane-2-ol, hexane -3-ol and cyclohexanol can be mentioned. Examples of lower heteroaliphatic alcohols include, but are not limited to, alkylene glycol monoalkyl ethers such as ethylene glycol monoalkyl ethers, propylene glycol monoalkyl ethers, and tetraglycols. In some specific embodiments, lower non-aromatic alcohols, such as lower aliphatic and lower heteroaliphatic alcohols, contain 1-8 non-hydrogen atoms, including any heteroatom.

Lower aromatic alcohols include lower alcohols with aromatic properties that can be contributed by aromatic (eg, phenyl, naphthyl, etc.) pendant groups on non-aromatic alcohols such as aliphatic alcohols or heteroaliphatic alcohols. Therefore, the alcohol group may be phenolic, primary, secondary, or tertiary. Examples of lower aromatic alcohols include, but are not limited to, phenols, benzyl alcohols, 2-methylbenzyl alcohols, and phenoxyethanol.

As used herein, "lower aliphatic diols" include saturated or unsaturated linear, branched, or cyclic aliphatic diols containing 2-10 carbon atoms. The alcohol groups may independently be primary, secondary, or tertiary to each other. In some specific embodiments, the lower aliphatic diol is a saturated or unsaturated linear or branched diol and is referred to herein as a "lower alkylene diol" (also referred to as a "lower glycol"). ). In some specific embodiments, the lower alkylene diol is a saturated linear or branched diol and is referred to herein as a "lower alkyl diol". In some specific embodiments, the lower alkyl diol is a linear diol and is referred to herein as a "lower n-alkyl diol". Specific examples of lower aliphatic diols useful in the solvent systems and gels described herein include, but are not limited to, ethane-1,2-diol (ethylene glycol), propane-1,2-diol (propylene). Glycol), propane-1,3-diol, butane-1,2-diol, butane-1,3-diol, butane-2,3-diol, butane-1,4-diol, pentane-1,5-diol , Pentane-1,2-diol, 2-methyl-2,4-pentanediol and the like.

As used herein, a "lower polyoxyalkylene" comprises a polymer formed from a lower aliphatic diol and having a molecular weight in the range of about 200 to about 600. Specific examples of lower polyoxyalkylenes useful in solvent systems and gels described herein include, but are not limited to, polyethylene glycols (eg, PEG 200, PEG 400, and PEG 600), polypropylene glycols (eg, PEG 600). , PPG-9), and dipropylene glycol.

The novel ternary solvent systems generally consist of about 3.5% to about 5% by weight of total lower aromatic alcohols (s) and about 95% to about 96.5% by weight of total lower aliphatic diols and polyoxyalkylenes. The weight ratio of total lower aliphatic diol: total polyoxyalkylene, including the mixture, is from about 1: 1 to about 1: 2, and in some specific embodiments it is about 1: 1.67.

The solvent system may include additional agents and solvents, such as additional agents or solvents that enhance the solubility of MTZ in aqueous solution, such as cyclodextrin, β-cyclodextrin, niacin, and / or niacinamide. However, high-dose MTZ aqueous gels that are homogeneous and have good storage stability properties can be prepared using solvent systems that do not contain such additional solubilizers, and in certain embodiments, lower aromatic alcohols. , Lower aliphatic diols, and lower polyoxyalkylenes only. Thus, in some particular embodiments, the solvent system and the resulting gel substantially contain agents that enhance the solubility of MTZ in aqueous solution, such as cyclodextrin, β-cyclodextrin, niacin, and / or niacinamide. Not included. In other specific embodiments, the solvent system is a ternary system consisting of one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more lower polyoxyalkylenes.

Solvent systems are generally from about 30% to about 60% by weight high dose mucosal adhesive MTZ aqueous gels, in certain embodiments from about 40% to about 50% by weight gels, and some specific embodiments. Represents a gel of about 40% to about 45% by weight, which may contain more or less total solvent.

Certain useful ternary solvent systems are benzyl alcohol as one or more lower aromatic alcohols, propane-1,2-diol (propylene glycol) as one or more lower aliphatic diols, and one or more lower polys. Contains polyethylene glycol having 400 MW (PEG 400) as oxyalkylene.

In some specific embodiments, a solvent system useful for dissolving MTZ in the high-dose mucosal-adhesive aqueous gels described herein is for the resulting high-dose mucosal-adhesive MTZ aqueous gel. , (A) Polyoxyalkylenes such as polyethylene glycol, for example PEG 400, from about 10% to about 50% by weight, for example, about 20% to about 40% by weight, or about 20% to about 30% by weight, In certain embodiments, it is contained in a concentration of about 25% by weight, (b) a lower aromatic alcohol such as phenoxyethanol or benzyl alcohol, and in certain embodiments, about 1.3% by weight to about 2.5% by weight, or benzyl alcohol. Containing from about 1.5% to 2.0% by weight, in certain embodiments at a concentration of about 2.0% by weight, and / or (c) lower aliphatic diols such as propane-1,2-diol, by more than about 3% by weight. Included in concentrations of ~ about 20% by weight, such as about 15% by weight to 20% by weight, and, in certain embodiments, about 15% by weight.

In yet other specific embodiments, the solvent system is (a) PEG 400, (b) PEG. 400 and lower aromatic alcohols such as benzyl alcohol, (c) PEG 400, lower aromatic alcohols such as benzyl alcohol and propane-1,2-diol, (d) lower aromatic alcohols such as benzyl alcohol, and / Or (e) contains propane-1,2-diol.

In yet other specific embodiments, the solvent system is (a) about 25% by weight PEG 400, (b) about 25% by weight PEG 400 and about 25% by weight of the resulting high dose MTZ mucosal adhesive aqueous gel. It contains 2% by weight benzyl alcohol, or (c) about 25% by weight PEG 400, about 2% by weight benzyl alcohol, and about 15% by weight propane-1,2-diol.

For gels designed for vaginal application, it is preferable to use a concentration of benzyl alcohol that does not cause irritation at the target site within the vaginal region. Thus, in some particular embodiments where the solvent system comprises benzyl alcohol, the amount of benzyl alcohol contained in the gel ranges from about 1.3% to about 2.5% by weight, for example from about 1.5% to about 2% by weight. In certain embodiments, it is about 2% by weight.

In yet another particular embodiment, the high dose mucosal adhesive MTZ aqueous gels described herein contain at least about 3% to about 20% by weight, for example about 15% by weight, of propane-1,2-diol. ~ About 20% by weight, including about 15% by weight in certain embodiments. In yet another particular embodiment, the gel comprises about 1.3% by weight MTZ and about 25% by weight PEG 400. In yet another particular embodiment, the gel comprises about 1.3% by weight MTZ, about 25% by weight PEG 400, and about 2% by weight benzyl alcohol. In yet another particular embodiment, the gel comprises about 1.3% by weight MTZ, about 25% by weight PEG 400, about 2% by weight benzyl alcohol, and about 15% by weight propylene glycol.

In yet another particular embodiment, the solvent systems useful for making the high dose mucosal adhesive MTZ aqueous gels described herein are (a) about 10% to about 50% by weight based on the gel. PEG 400 by weight, eg, about 20% to about 40% by weight, or about 20% to about 30% by weight, in certain embodiments about 25% by weight, (b) about 1.3% by weight. Approximately 2.5% by weight, eg, about 1.5% to about 2% by weight, in certain embodiments, about 2% by weight of benzyl alcohol, (c) at least about 3% to about 20% by weight, such as about 15% by weight. About 20% by weight, in certain embodiments, about 15% by weight of propane-1,2-diol, (d) about 25% by weight PEG 400, (e) about 25% by weight PEG 400 and about 2. It contains one of (f) about 25% by weight PEG 400, about 2% by weight benzyl alcohol, and about 15% by weight propylene glycol.

The pH of the high-dose mucosal adhesive MTZ aqueous gels described herein should generally match the pH of the intended application region, eg, for a healthy vaginal application when intended for vaginal application. Must match pH. Therefore, for gels intended for vaginal application, the pH should generally be in the range of about pH 3 to about pH 5, for example about pH 4. The pH can be adjusted and / or maintained using acids, base buffers, and other pH adjusters, well known in the art and discussed in the summary section.

The high-dose mucosal adhesive MTZ aqueous gels described herein can also contain other additional components, such as one or more preservatives, as is well known in the art. When used, preservatives (s) generally contain only about 1% by weight or 2% by weight of high dose mucosal adhesive MTZ aqueous gels, typically as high as about 0.25% by weight to about 1.0% by weight. Dose Includes Mucosal Adhesive MTZ Aqueous Gel. The choice of preservative (s) is not important. Many preservatives suitable for use in pharmaceutical formulations are well known to those of skill in the art. Any of these preservatives, and in some specific embodiments, those having antibacterial properties, used alone or in combination in the high dose mucosal adhesive MTZ aqueous gels described herein. Can be done. In some specific embodiments, one or more preservatives are esters of 4-hydroxybenzoic acid, also known as parabens. Suitable parabens include lower alkyl esters of 4-hydroxybenzoic acid, such as methyl 4-hydroxybenzoate (methylparaben), ethyl 4-hydroxybenzoate (ethylparaben), and propyl4-hydroxy-benzoate (propylparaben). ..

Experts will understand that the solvent used to dissolve the MTZ in the high dose mucosal adhesive MTZ aqueous gels described herein may have storage properties. For example, benzyl alcohol has well-known storage properties. Preservation properties can be used advantageously when used as a solvent in a new solvent system. In fact, gels containing solvents with preservative properties do not need to contain additional preservatives. Gels that utilize the storage properties of the solvent contained in the solvent system provide effective storage properties after a desired period of time in addition to the MTZ dissolution properties that the gel is effective in, in examples where the solvent can decompose and / or oxidize over time. It should contain an amount of excess that takes into account decomposition and / or oxidation so as to retain some amount of undifferentiated and / or unoxidized solvent. For example, benzyl alcohol is known to oxidize to benzaldehyde, which has no storage properties. In embodiments of high-dose MTZ gels described herein using benzyl alcohol as an MTZ solvent and as a preservative, certain amounts of benzyl alcohol not only dissolve the MTZ, but also the expected storage of the gel. Must be included in a gel that provides a shelf life for a period of time. The excess amount of benzyl alcohol, or any other solvent used as a partial preservative in the solvent system, can be determined based on the degradation and / or oxidation properties of the particular solvent, as well as its kinetics, under the desired storage conditions.

Embodiments of high-dose mucosal adhesive MTZ aqueous gels containing solvents with preservative properties include one or more additional preservatives and / or preservatives designed to protect the solvent from decomposition and / or oxidation. May include. For example, in the case of benzyl alcohol, the high dose mucosal adhesive MTZ aqueous gels described herein provide one or more additional preservatives with antioxidant properties to protect some benzyl alcohol from oxidation. May include. In certain embodiments, the high dose mucosal adhesive MTZ aqueous gel containing benzyl alcohol comprises one or more parabens as an additional preservative. In some specific embodiments, the high dose mucosal adhesive MTZ aqueous gel has about 0.1% by weight of total parabens, and in some specific embodiments, about 0.02% by weight of methylparaben and about 0.08% by weight of propyl. Including parabens.

The high dose mucosal adhesive MTZ aqueous gel may contain water in either the form of pure water or in the form of an aqueous buffer. Typically, the amount of water contained in the gel is less than about 70% by weight, more typically less than about 60% by weight, and in some specific embodiments from about 45% to about 55% by weight. The range.

Certain exemplary high-dose mucosal adhesive MTZ aqueous gels (a) At least 1.3% by weight MTZ, about 1.2% to about 2% by weight MTZ, about 3% to about 1.5% by weight MTZ, or about 1.3% by weight MTZ, (b) With one or more mucosal adhesive gelled polymers, including.

Another specific exemplary high-dose mucosal adhesive MTZ aqueous gel (a) MTZ from about 1.2% to about 2% by weight, such as about 1.3% to about 1.5% by weight MTZ, and in certain embodiments about 1.3% by weight MTZ. (b) With one or more mucosal adhesive gelled polymers, (i) Approximately 25% by weight PEG 400 and / or (ii) Approximately 2% by weight benzyl alcohol and / or (iii) Approximately 15% by weight propane-1,2-diol including.

Another specific exemplary high-dose mucosal adhesive MTZ aqueous gel (a) Approximately 1.3% by weight MTZ and 25% by weight PEG 400, (b) (a) Approximately 1.3% by weight MTZ, approximately 25% by weight PEG 400, approximately 2% by weight benzyl alcohol, or (c) Approximately 1.3% by weight MTZ, approximately 25% by weight PEG 400, approximately 2% by weight benzyl alcohol, and approximately 15% by weight propane-1,2-diol. including. 6.3. How to make a gel

The high-dose mucosal-adhesive MTZ aqueous gel described herein generally comprises dissolving the water-soluble component of the gel in water or a buffer to produce an aqueous solution, and mixing the MTZ solvent-based components to mix the MTZ solvent-based It can be prepared by dissolving the desired amount of MTZ in it to produce an MTZ solution, mixing the required amount of aqueous solution and MTZ solution together, and adding the desired amount of gelling agent to the mixture. Depending on the gelled polymer used, the pH of the MTZ solution should be adjusted to a certain range with an acid and / or base prior to addition to the gelled polymer (s) and then the pH of the resulting gel should be adjusted. It may be desirable to adjust to the desired specific range with an acid or base. A specific method for making high-dose mucosal adhesive MTZ aqueous gels is provided in Example 2. 6.4. Use

The high-dose mucosal adhesive MTZ aqueous gels described herein can be used to administer MTZ topically in any context in which such administration is advantageous. In certain embodiments further described below, the gel can be advantageously used intravaginally to treat women with or diagnosed with BV.

As used herein, if a woman experiences one or more of the symptoms, conditions, or presentations that are found to be associated with BV, she is said to be "affected" or diagnosed with BV. It is said. Individuals with BV typically present with a particularly unpleasant "fishy" off-white, thin, homogeneous vaginal discharge and no inflammatory response. Individuals have a decrease in the spread and concentration of Lactobacillus (particularly the form that produces hydrogen peroxide), as well as the accompanying Gardnerella vaginalis, Mobiluncus spp, (Bacteroid, Prebotera, and Porphyro). It also presents with an increase in anaerobic gram-positive rods (of the genus Mobiluncus), Peptostreptococcus spp, and Mycoplasma hominis. Predispositions are non-white species, pre-pregnancy, IUD use, sexual activity, new sexual partners, and recent antibiotic use. BV is also associated with trichomoniasis.

Criteria for establishing a clinical diagnosis of BV are provided, in particular, in the draft FDA Guidance entitled "Guidance for Industry: Bacterial Vaginosis-Developing Antimicrobial Drugs for Treatment" dated July 1998. The whole is incorporated herein (hereinafter referred to as "FDA Guidance"). As per FDA guidance, the clinical diagnosis of BV includes the following observations: 1. Off-white (milky or gray) thin homogeneous secretions with or without vulvar and vaginal pruritus and inflammation 2. Presence of "filamentous cells" in 20% or more of total epithelial cells during microscopic examination of saline "wet mount" 3. Vaginal discharge above pH 4.5, and 4. Vaginal secretion fish odor when 10% KOH is added dropwise (ie, "odor test" positive).

These observations are commonly referred to as the "Amcel criteria" and women who present all four criteria are referred to herein as "Amcel positive".

Women who experience vaginitis, which is considered to be BV, typically also have a Gram-stained slide Nugent score of 4 or higher. The Nugent score is based on the total weight of the following three bacterial polymorphisms calculated from a slide review under oil immersion at a 1000-fold magnification. . Lactic acid bacteria: large gram-positive bacilli . Gardonerella / Bacteroid: Small gram indefinite bacilli / Small gram negative bacilli Mobiluncus: a thin curved gram indefinite bacillus

BV criteria that comply with the Nugent criteria are a total score of 7 or higher, scores 4-6 are considered intermediate, and scores 0-3 are considered normal. A score of 3 or higher is considered abnormal by the FDA.

Morphological types are scored as the average number found per oil-soaked area (minimum 10-20 areas should be examined). Next, each form type is given a numerical score as follows. 0 = No polymorphisms seen, 1 + = 1 less than 1 polymorphism per region, 2 + = 1 ~ 4 polymorphisms per region, 3 + = 1 5-30 polymorphisms per region, and 4 + = 1 More than 30 polymorphisms per area. The total Nugent score is calculated by summing the scores of the individual polymorphisms. For more information on the Nugent scoring system, see Nugent et al., 1991, Reliability of Diagnosing Bacterial Vaginosis is Improved by a Standardized Method of Gram Stain Interpretation, J. See Clin Micrbiol 29 (2): 297-301. Women with a Nugent score greater than 4 are referred to herein as "Nugent Positive".

In some embodiments, individuals presenting at least three of the Amsel criteria described above are considered to have and / or have been diagnosed with BV. In some embodiments, Nugent-positive individuals are considered to have and / or have been diagnosed with BV. In certain embodiments, individuals suffering from and / or being diagnosed with BV are Amsel positive. In yet other specific embodiments, individuals suffering from, and diagnosed with, BV are both Amsel-positive and Nugent-positive.

Preferably, the individual suffering from and / or being diagnosed with BV does not have other vaginal infections including, but not limited to, Chlamydia, Trichomonas, gonorrhea, and Candida. Preferably, individuals suffering from and / or being diagnosed with BV have not been treated for other vaginal infections including, but not limited to, Chlamydia, Trichomonas, gonorrhea, and Candida. In one embodiment, an individual suffering from and / or being diagnosed with BV is being treated with fluconazole.

This method generally provides the therapeutic benefit to women suffering from and / or being diagnosed with BV a certain amount of the high dose mucosal adhesive MTZ aqueous gel described herein. It involves vaginal application with a sufficient number of applications. As used herein, a "therapeutic benefit" is one or more of the symptoms of BV and / or one or more of the clinical signs of BV, including, for example, one or more of the Amsel criteria or Nugent scores. Refers to mitigation, extinction, eradication, or improvement. Treatment plans can provide therapeutic benefits to a subject without curing the underlying BV condition.

As used herein, symptoms or signs of BV are alleviated when the degree after treatment diminishes or approaches levels considered normal. For example, the Amsel criterion is that if vaginal discharge returns to normal, the number of filamentous cells in the wet mount is less than about 20% of vaginal epithelial cells, eg, about 19%, 18%, 17%, 16%. , 15%, 10%, less than 5%, or even less, the pH of vaginal discharge is from about pH 4.7 to about pH 5.3 before treatment to about pH 4.0 to about pH 4.5 after treatment. Improved within the range, eg, when reduced to pH4.3, pH4.2, pH4.1, or pH4.0, and / or when the odor test does not produce large amounts of amine or "fishy odor". Symptomatology of BV is eliminated or eradicated when the symptoms are no longer detected using well-known detection means (eg, those disclosed in the various detection means provided in the Examples section and FDA guidance). It is thought that it was.

As used herein, "therapeutically effective amount" refers to a treatment scheme with a high-dose mucosal adhesive MTZ aqueous gel that provides therapeutic benefits.

In some embodiments, an amount of high dose mucosal adhesive MTZ aqueous gel shows improvement in at least one Amsel criterion, preferably at least 2 or 3, and most preferably all four Amsel criteria, and / or Applies enough times to result in a Nugent score of less than 4.

In some specific embodiments, a certain amount of high dose mucosal adhesive MTZ aqueous gel is applied with sufficient number of applications to result in clinical healing. As used herein, "clinical cure" is achieved if all four Amsel criteria are neutralized as described below. 1. The original secretory properties of BV have returned to normal physiological secretion, which varies in appearance and consistency with the physiological cycle. 2. Saline wet mount is negative for filamentous cells. 3. The pH of vaginal discharge is less than 4.7 pH and is typically measured using pH paper measuring pH 4.0-pH 6.0. 4. The odor test is negative for any amine ("fish") odor.

In some specific embodiments, a certain amount of high dose mucosal adhesive MTZ aqueous gel is applied with sufficient number of applications to result in bacteriological healing. As used herein, "bacteriological cure" results in a Nugent score of less than 4, for example, a Nugent score in the range of 1, 2, or 3, or 0-3. Sometimes achieved.

In yet other specific embodiments, certain amounts of high dose mucosal adhesive MTZ aqueous gels described herein are applied in sufficient number of applications to result in therapeutic healing. As used herein, "therapeutic cure" is achieved when both clinical and bacteriological cures are obtained.

In yet another particular embodiment, a certain amount of high dose mucosal adhesive MTZ aqueous gel is applied with sufficient number of applications for the physician to determine that no additional treatment is required. In yet another specific embodiment, certain amounts of high-dose mucosal adhesive MTZ aqueous gels described herein are applied in sufficient number of applications to achieve any healing level as defined in the FDA Guidance. Will be done.

In some specific embodiments, the amount of high dose mucosal adhesive MTZ aqueous gel applied in a single application contains from about 60 mg to about 100 mg MTZ. In some specific embodiments, the amount of high dose mucosal adhesive MTZ aqueous gel applied in a single application contains from about 60 mg to about 80 mg, or from about 60 mg to about 70 mg of MTZ. In some specific embodiments, a high dose MTZ aqueous gel applied in a single application in an amount contains about 65 mg of MTZ.

The frequency and duration of application may vary and may depend on the desired outcome. Generally, the gel is applied once daily for a period of 1, 2, 3, 4, or 5 days. A single application of a high-dose mucosal adhesive MTZ aqueous gel containing approximately 65 mg of MTZ was found to achieve significant therapeutic benefits. Therefore, in some embodiments, the gel is applied in a single application, i.e. without the additional application of a high dose mucosal adhesive MTZ aqueous gel. Application of high-dose mucosal adhesive MTZ aqueous gel containing approximately 65 mg MTZ, applied once daily for a 5-day period, is lower than a similar course of treatment with FDA-approved 0.75 wt% metronidazole gel. It was also found that there was virtually no occurrence of vulvar vaginal candidiasis after treatment, resulting in a recurrence rate. Therefore, in some embodiments, the gel is applied once daily for a period of 5 days.

As used herein, the term "recurrence" means that after treatment with the high-dose MTZ gels described herein, at least one of the symptoms of BV as defined herein reappears and the end of treatment. It means that the subject will later be diagnosed with at least a second BV.

"Reduced recurrence rate" is due to the high dose MTZ gels described herein as compared to the time to historical recurrence observed with conventional 0.75 wt% MTZ gels such as METROGEL VAGINAL®. It means an increase in the time to recurrence of BV symptoms after the end of treatment. In certain embodiments, the reduction in recurrence rate means that the symptoms of BV are at least about 19 days after the end of treatment, eg, 20, 21, 22, 23, 24, 25, 26, 27. It means that it will not be allowed for a period of days, 28 days, 29 days, or more. BV symptoms may not recur for at least 29 days after the end of treatment, eg, 30, 35, 40, 45, 50, 3 months, 6 months, 9 months, 1 year, or longer. , Suggesting that there is no recurrence in the subject after the end of treatment.

"Reduced recurrence rate" can also be defined for the study population. For example, the high-dose mucoadhesive MTZ aqueous gel treatment regimen described herein, which statistically significantly reduces the number of women experiencing BV symptoms more than 20 days after treatment, is 0.75 weight by weight. The recurrence rate of BV is considered to be reduced compared to the number of people who reported symptoms of BV more than 20 days after treatment with% MTZ gel (eg, METROGEL VAGINAL®).

For use, the high dose mucosal adhesive MTZ aqueous gel can be packaged in any form that is convenient for the desired application form. In certain embodiments useful for vaginal application, the high dose mucosal adhesive MTZ aqueous gel is packaged in unit dose form, as a particular example, as a prefilled dose syringe type applicator. 6.5. Additional non-limiting aspects

The components of the high-dose mucosal adhesive MTZ aqueous gel, as well as additional non-limiting aspects of the methods used to treat women suffering from and / or being diagnosed with BV, are provided below. .. 6.5.1. Method

A method of treating a subject suffering from and / or being diagnosed with M1.BV, one or more mucosal adhesive gelled polymers, about 1% to about 2% by weight of metronidazole (MTZ), And a method comprising intravaginal application of a mucosal adhesive aqueous gel containing water, wherein a certain amount of mucosal adhesive aqueous gel applied in a single application contains from about 60 mg to about 100 mg of MTZ.

M2. In an in vitro Franzsel skin permeation experiment performed on human corpse skin, when the level of MTZ from the mucosal adhesive aqueous gel measured in the stratum corneum and the receptor was normalized for concentration. The method according to embodiment M1, which is at least about 25 to 55 times higher and at least about 1 to 20 times lower than the MTZ level measured from METROGEL VAGINAL®, respectively.

M3. Mucoadhesive aqueous gel has viscosities ranging from about 200,000 mPa to about 400,000 mPa measured at 25 ° C using controlled shear rate method, Bohlin CVO100 rheometer, and rheometer settings in Table 11. , The method according to aspect M1.

M4. The method of embodiment M1, having a degree of mucosal adhesion within about ± 10% of that of MG33PB.

M5. The method according to embodiment M1, wherein the mucosal adhesive aqueous gel comprises from about 30% to about 60% by weight of water.

M6. The method of embodiment M1, wherein the mucosal adhesive aqueous gel is substantially free of dextrin, cyclodextrin, niacin, and niacinamide.

M7. The method according to embodiment M1, wherein the mucosal adhesive aqueous gel is stable at 25 ° C for a period of at least 6 months.

M8. Any one of embodiments M1 to M7, wherein the mucosal adhesive aqueous gel comprises from about 1% to about 3% by weight total mucosal adhesive gel polymer and from about 40 to 45% by weight of MTZ solvent system. The method described in.

M9. The method of embodiment M8, wherein the solvent system comprises one or more solvents having a saturated MTZ solubility of at least about 20 mg / g at 25 ° C.

M10. The solvent system is one or more solvents with a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and optionally a saturated MTZ solubility in the range of about 20 mg / g to about 25 mg / g at 25 ° C. The method of aspect M9, comprising one or more solvents having.

M11. The solvent system is one or more lower aromatic alcohols, and optionally one or more lower aliphatic diols, and / or one or more polyoxyalkylenes having a molecular weight in the range of about 200 to about 400 ("" The method of embodiment M10, comprising "lower polyoxyalkylene").

M12. The method of embodiment M11, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and / or one or more lower polyoxyalkylenes.

M13. The method of embodiment M12, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more lower polyoxyalkylenes.

M14. The method according to any one of aspects M11 to M13, wherein one or more lower aromatic alcohols are selected from the group consisting of benzyl alcohol, phenoxyethanol, and mixtures thereof.

M15.1 One or more lower alkylene diols are selected from the group consisting of ethane-1,2-diol (ethylene glycol), propane-1,2-diol (propylene glycol), and mixtures thereof, embodiments M11 ~. The method according to any one of M14.

M16.1 In any one of aspects M11 to M15, one or more lower polyoxyalkylenes are selected from the group consisting of polyoxyethylene (polyethylene glycol), polyoxypropylene (polypropylene glycol), and mixtures thereof. The method described.

M17.1 One or more lower aliphatic diols and one or more lower polyoxyalkylenes in the total lower aliphatic diol: total lower polyoxyalkylene weight ratio in the range of about 1: 1 to about 1: 2 said solvent. The method according to any one of aspects M11 to M16, which is included in the system.

M18. Solvent system with about 3.5% to about 5% by weight of total lower aromatic alcohols and about 95% to about 95.5% by weight of one or more lower alkylene diols and one or more lower polyoxyalkylenes. The method according to any one of aspects M13 to M17, comprising a mixture of.

M19. The method of aspect M18, wherein the weight ratio of total lower alkylene diol: total lower polyoxyalkylene is in the range of about 1: 1 to about 1: 1.67.

M20. Solvent system is benzyl alcohol, propane-1,2-diol, and PEG The method according to any one of aspects M8 to M19, comprising 400.

M21. The solvent system consists of about 3.5% to about 5% by weight benzyl alcohol and about 95% to about 96.5% by weight of a mixture of propane-1,2-diol and PEG 400. The method according to any one of aspects M8 to M20, wherein the weight ratio of 2-diol: PEG 400 is in the range of about 1: 1 to about 1: 1.67.

M22. The method according to any one of aspects M1 to M21, wherein the mucosal adhesive aqueous gel further comprises one or more preservatives.

M23. The method according to any one of aspects M1 to M22, wherein the preservative is an ester of 4-hydroxybenzoic acid (paraben).

M24. The method of embodiment M23, wherein one or more preservatives are selected from the group consisting of methyl 4-hydroxybenzoate (methylparaben), propyl4-hydroxybenzoate (propylparaben), and mixtures thereof.

M25. The method according to any one of aspects M1 to M23, wherein the mucosal adhesive aqueous gel has a pH in the range of about pH 3.0 to about pH 5.0.

M26. The method of embodiment M25, wherein the mucosal adhesive aqueous gel has a pH of about pH 4.0.

M27. The method according to any one of aspects M1 to M26, wherein the mucosal adhesive gelled polymer is selected from the group consisting of hydroxyethyl cellulose, carbomer, polycarbophil, and mixtures thereof.

M28. The method of any one of embodiments M1 to M27, wherein the amount of mucosal adhesive aqueous gel applied in a single application contains approximately 65 mg of MTZ.

M29. The method according to any one of aspects M1 to M28, wherein the mucosal adhesive aqueous gel comprises from about 1% to about 1.5% by weight of MTZ.

M30. The method of embodiment M29, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 1-5 days.

M31. The method of embodiment M29, wherein the mucosal adhesive aqueous gel is applied once daily for a period of one day.

M32. The method of embodiment M29, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days.

M33. The method according to any one of aspects M1 to M28, wherein the mucosal adhesive aqueous gel comprises about 1.3% by weight of MTZ.

M34. The method of embodiment M33, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 1-5 days.

M35. The method of embodiment M33, wherein the mucosal adhesive aqueous gel is applied once daily for a period of one day .

M36. The method of embodiment M33, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days.

M37. Mucoadhesive aqueous gel, about 1.3% by weight MTZ, about 2% by weight polycarbophil AA-1, about 2% by weight benzyl alcohol, about 15% by weight propane-1,2-diol, about The method according to any one of aspects M1 to M7, comprising 25% by weight PEG400, about 0.02% by weight methyl 4-hydroxybenzoate, and about 0.08% by weight propyl4-hydroxybenzoate.

M38. The method of embodiment M37, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 1-5 days.

M39. The method of embodiment M37, wherein the mucosal adhesive aqueous gel is applied once daily for a period of one day.

M40. The method of embodiment M37, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days.

M41. The method of any one of embodiments M37-M40, wherein the amount of mucosal adhesive aqueous gel applied in a single application contains approximately 65 mg of MTZ.

A method of treating subjects with and / or diagnosed with M42.BV, which is applied intravaginally to a certain amount of aqueous gel in sufficient number of applications to achieve clinical cure. The aqueous gel contains one or more mucosal adhesive polymers, about 1.3% by weight MTZ, and water. (a) When the levels of MTZ from mucosal adhesive aqueous gels measured in the stratum corneum and in the receptor are normalized for concentration in in vitro Franzsel skin permeation experiments performed on human corpse skin. , At least about 25-55 times higher and at least about 1-20 times lower than the MTZ levels measured from METROGEL VAGINAL®, respectively. (b) Viscosities in the range of about 200,000 mPa to about 400,000 mPa, measured at 25 ° C, using the controlled shear rate method, the Bohlin CVO100 rheometer, and the rheometer settings in Table 11. (c) Degree of mucosal adhesion within about ± 10% of that of MG33PB, (d) Containing about 30% by weight to about 60% by weight of water, (e) Substantially free of dextrin, cyclodextrin, niacin, and niacin amide, (f) Stable at 25 ° C for at least 6 months, (g) Containing about 1% to about 3% by weight of total mucosal adhesive polymer and about 40% to 45% by weight of MTZ solvent system, A method having one or more features or properties selected from the group of.

M43. The method of embodiment M42, wherein an amount of aqueous gel is applied a sufficient number of applications to achieve therapeutic healing.

A method of treating women with and / or diagnosed with M44.BV, one or more mucoadhesive gels of about 1.3% by weight MTZ, about 1% to about 2% by weight. A single application of a mucosal adhesive MTZ aqueous gel containing a polymer, about 40% to about 50% by weight of MTZ solvent, and about 50% to about 55% by weight of water can be applied intravaginally to women. Containing, the solvent system is about 3% to about 5% by weight of one or more solvents having a saturated MTZ solubility of at least about 50 mg / g at 25 ° C, and about 20 mg / g to about 25 mg / g at 25 ° C. The amount of mucoadhesive MTZ aqueous gel applied, containing one or more solvents from about 95% to about 97% by weight, collectively having saturated MTZ solubility in the range of g, is from about 60 mg to about 100 mg. A method containing MTZ.

M45. The method of embodiment M44, wherein the applied amount of mucosal adhesive aqueous gel contains approximately 65 mg of MTZ.

46. The method of any one of embodiments M43-M45, wherein the solvent is benzyl alcohol with a saturated MTZ solubility of at least 50 mg / g at 25 ° C.

One or more solvents collectively having saturated MTZ solubilities in the range of about 20 mg / g to about 25 mg / g at M47.25 ° C are selected from the group consisting of lower aliphatic diols and lower polyoxyalkylenes, respectively. The method according to any one of aspects M43 to M46.

M48. Lower aliphatic diols in which the solvent system comprises from about 3% to about 5% by weight benzyl alcohol and from about 95% to about 97% by weight a mixture of lower aliphatic diols and lower polyoxyalkylenes: The method according to any one of aspects M43 to M47, wherein the weight ratio of the lower polyoxyalkylene is from about 1: 1 to about 1: 2.

M49. The method of embodiment M48, wherein the lower aliphatic diol: lower polyoxyalkylene has a weight ratio of about 1: 1.67.

M50. The method according to any one of aspects M48 to M49, wherein the lower aliphatic diol is propane-1,2-diol and the lower polyoxyalkylene is PEG 400.

M51. The method according to any one of aspects M43 to M50, wherein one or more mucosal adhesive gelled polymers are selected from the group consisting of crosslinked acrylic acid polymers and cellulose polymers, respectively.

M52. The method according to any one of aspects M43 to M51, wherein one or more mucosal adhesive gelled polymers are selected from the group consisting of carbomer and polycarbophil, respectively.

M53. The method according to any one of aspects M43 to M52, wherein the mucosal adhesive gelled polymer is polycarbophil.

M54. Mucoadhesive MTZ aqueous gel, about 1.3% by weight MTZ, about 2% by weight polycarbofyl AA-1, about 2% by weight benzyl alcohol, about 15% by weight propane-1,2-diol, The method of any one of embodiments M43-M53, comprising about 25% by weight PEG400, about 0.1% by weight one or more preservatives, and about 54.6% by weight water.

M55. The method of embodiment M54, wherein each of the two or more preservatives is paraben.

M56. The method of embodiment M55, wherein the one or more preservatives are methylparaben and propylparaben.

M57. Mucoadhesive MTZ aqueous gel, about 1.3% by weight MTZ, about 2% by weight polycarbofyl AA-1, about 2% by weight benzyl alcohol, about 15% by weight propane-1,2-diol, The method according to any one of aspects M43 to M56, comprising about 25% by weight PEG400, about 0.08% by weight methylparaben, about 0.02% by weight propylparaben, and about 54.6% by weight water.

A method of treating a subject suffering from and / or being diagnosed with M58.BV with a single application of a composition suitable for it, one or more mucosal adhesive gelled polymers, water, and. The amount of mucosal adhesive aqueous gel applied in a single application, including intravaginal application to a mucosal adhesive aqueous gel containing about 1.3% by weight of MTZ, contains about 65 mg of MTZ. A method in which a single application is sufficient to treat or cure BV in the complete or substantial absence of further treatment or application.

A method of treating a subject suffering from and / or being diagnosed with M59.BV, including intravaginal application of a single dose of mucosal adhesive aqueous MTZ gel to the subject, mucosal adhesive aqueous MTZ. The gel contains one or more mucosal adhesive gelled polymers, about 1.3% by weight MTZ, and water, and the amount of mucosal adhesive aqueous gel applied in a single dose contains about 65 mg MTZ. Method.

A method of treating a subject suffering from and / or being diagnosed with M60.BV, which comprises intravaginally applying a mucosal adhesive aqueous MTZ gel to the subject, (i) a mucosal adhesive aqueous MTZ gel. However, it contains one or more mucosa-adhesive gelled polymers, about 1.3% by weight of MTZ, and water, and (ii) the amount of mucosa-adhesive aqueous MTZ gel applied contains about 65 mg of MTZ, ( iii) A method in which a mucosal adhesive aqueous MTZ gel is applied once without further application.

A method of treating a subject suffering from and / or being diagnosed with M61.BV, which comprises intravaginally applying a mucosal adhesive aqueous MTZ gel to the subject, (i) a mucosal adhesive aqueous MTZ gel. Contains one or more mucosal adhesive gelled polymers, about 1.3% by weight MTZ, and water, (ii) the amount of mucosal adhesive aqueous gel applied in a single application contains about 65 mg MTZ. And (iii) a method in which a mucosal adhesive aqueous MTZ gel is applied once a day for a period of one day.

A method of treating subjects with and / or diagnosed with M62.BV, including intravaginal application of mucosal adhesive aqueous MTZ gels to subjects once daily for a period of 5 days. (i) Mucosal adhesive aqueous MTZ gel contains one or more mucosal adhesive gelled polymers, about 1.3% by weight MTZ, and water, and (ii) mucosal adhesive aqueous MTZ gel applied in a single application. The method, the amount of which contains about 65 mg of MTZ.

M63. A method of treating subjects with and / or diagnosed with BV, vaginal application to a single application of a sufficient amount of mucosal adhesive aqueous MTZ gel to cure BV. A method in which a mucosal adhesive aqueous MTZ gel comprises one or more mucosal adhesive gelled polymers, about 1.3% by weight of MTZ, and water.

M64. The method according to any one of aspects M58 to M63, wherein the mucosal adhesive aqueous MTZ gel is the gel according to any one of aspects C1 to 32 below. 6.5.2. Composition

C1. About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of MTZ solvent system, In an in vitro Franzcel skin permeation experiment performed with human corpse skin containing and water, the level of MTZ from mucosal adhesive aqueous gels measured in the stratum corneum and in the receptor was normalized for concentration. Mucosal adhesive aqueous gels, when, at least about 25-55 times higher and at least about 1-20 times lower than the MTZ levels measured from METROGEL VAGINAL®, respectively.

C2. About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of MTZ solvent system, Mucosal adhesions containing water and with viscosities ranging from about 200,000 mPa to about 400,000 mPa measured at 25 ° C using a controlled shear rate method, the Bohlin CVO100 rheometer, and the rheometer settings in Table 11. Rheometer gel.

C3. About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of MTZ solvent system, And a mucoadhesive aqueous gel containing water and having a degree of mucosal adhesion within about ± 10% of that of MG33PB.

C4. About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of MTZ solvent system, And a mucoadhesive aqueous gel containing about 30% to about 60% by weight of water.

C5. About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of MTZ solvent system, A mucosal adhesive aqueous gel containing water and substantially free of dextrin, cyclodextrin, niacin, and niacinamide, and optionally a surfactant.

C6. About 1% to about 2% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, about 30% to about 60% by weight of MTZ solvent system, And a mucoadhesive aqueous gel containing water and stable at 25 ° C for a period of at least 6 months.

C7. Any one of aspects C1 to C6, wherein the mucosal adhesive aqueous gel comprises from about 1% to about 3% by weight of total mucosal adhesive gel polymer and from about 40 to 45% by weight of MTZ solvent system. The gel described in.

C8. The gel according to embodiment C7, wherein the solvent system comprises one or more solvents having a saturated MTZ solubility of at least about 20 mg / g at 25 ° C.

C9. The solvent system is saturated at least about 50 mg / g at 25 ° C. One or more solvents with MTZ solubility and / or optionally saturation in the range of about 20 mg / g to about 25 mg / g at 25 ° C. The gel according to embodiment C8, which comprises one or more solvents having MTZ solubility.

C10. The solvent system is one or more lower aromatic alcohols, and optionally one or more lower aliphatic diols, and / or one or more polyoxyalkylenes having a molecular weight in the range of about 200 to about 400 ("" The gel according to aspect C9, comprising "lower polyoxyalkylene").

C11. The gel according to embodiment C10, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and / or one or more lower polyoxyalkylenes.

C12. The gel according to embodiment C11, wherein the solvent system comprises one or more lower aromatic alcohols, one or more lower aliphatic diols, and one or more lower polyoxyalkylenes.

C13. The gel according to any one of aspects C10 to C12, wherein one or more lower aromatic alcohols are selected from the group consisting of benzyl alcohol, phenoxyethanol, and mixtures thereof.

C14. One or more lower alkylene diols are selected from the group consisting of ethane-1,2-diol (ethylene glycol), propane-1,2-diol (propylene glycol), and mixtures thereof, embodiments C10 ~ The gel according to any one of C13.

C15.1 In any one of aspects C10 to C14, one or more lower polyoxyalkylenes are selected from the group consisting of polyoxyethylene (polyethylene glycol), polyoxypropylene (polypropylene glycol), and mixtures thereof. The gel described.

C16.1 One or more lower aliphatic diols and one or more lower polyoxyalkylenes are solvent-based in a total lower aliphatic diol: total lower polyoxyalkylene weight ratio in the range of about 1: 1 to about 1: 2. The gel according to any one of aspects C10 to C15, which is contained in.

C17. Solvent system with about 3.5% to about 5% by weight of total lower aromatic alcohols and about 95% to about 95.5% by weight of one or more lower alkylene diols and one or more lower polyoxyalkylenes. The gel according to any one of aspects C10 to C16, which comprises a mixture of.

C18. The gel according to embodiment C17, wherein the total lower alkylene diol: total lower polyoxyalkylene has a weight ratio of about 1: 1.67.

C19. Solvent systems are benzyl alcohol, propane-1,2-diol, and PEG The gel according to any one of aspects C10 to C18, which comprises 400.

C20. The solvent system consists of about 3.5% to about 5% by weight benzyl alcohol and about 95% to about 96.5% by weight of a mixture of propane-1,2-diol and PEG 400. The gel according to any one of aspects C10 to C19, wherein the weight ratio of 2-diol: PEG 400 is in the range of about 1: 1 to about 1: 1.67.

C21. The gel according to any one of aspects C1 to C20, further comprising one or more preservatives.

C22. The gel according to any one of aspects C1 to C21, wherein the preservative is an ester of 4-hydroxybenzoic acid (paraben).

C23. The gel according to embodiment C22, wherein one or more preservatives are selected from the group consisting of methyl 4-hydroxybenzoate (methylparaben), propyl4-hydroxybenzoate (propylparaben), and mixtures thereof.

C24. The gel according to any one of aspects C1 to C23, having a pH in the range of about pH 3.0 to about pH 5.0.

C25. The gel according to embodiment C24, having a pH of about pH 4.0.

C26. The gel according to any one of aspects C1 to C25, wherein one or more mucosal adhesive gelled polymers are selected from the group consisting of hydroxyethyl cellulose, carbomer, polycarbophil, and mixtures thereof.

C27. Mucoadhesive aqueous gel, about 1.3% by weight MTZ, about 2% by weight polycarbofil AA-1, about 2% by weight benzyl alcohol, about 15% by weight propane-1,2-diol, about The gel according to any one of aspects C1 to C6, comprising 25% by weight PEG400, about 0.02% by weight methyl 4-hydroxybenzoate, and about 0.08% by weight propyl4-hydroxybenzoate.

C28. Solvent system containing about 1.3% by weight of MTZ, about 1% to about 3% by weight of one or more mucosal adhesive gelled polymers, and about 40% to about 45% by weight of MTZ solvent system. However, when the mucosal adhesive aqueous gel contains benzyl alcohol, propane-1,2-diol, and PEG400 and is stored at a temperature in the range of about 25 ° C to about 40 ° C, a period of 18 months. A mucosal adhesive aqueous gel that is stable during.

C29. The gel according to embodiment C28, wherein one or more mucosal adhesive gelled polymers are selected from the group consisting of crosslinked acrylic acid polymers and cellulose polymers.

C30. The gel according to embodiment C29, wherein the crosslinked acrylic acid polymer is selected from the group consisting of carbomer and polycarbophil.

C31. The gel according to any one of aspects C29 to C30, wherein the cellulose polymer is selected from the group consisting of hydroxyethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose.

C32. The gel according to any one of aspects C29 to C31, comprising about 2% by weight of total mucosal adhesive gelled polymer. 6.5.3. Use

Use of the mucosal adhesive aqueous gel according to any one of embodiments C1-C32 for the treatment of women suffering from and / or being diagnosed with U1.BV.

U2. Use of embodiment U1, in which the mucosal adhesive aqueous gel is applied intravaginally with sufficient application times to provide bacteriological, clinical, or therapeutic cure.

U3. The use according to any one of aspects U1 to U2, wherein the amount of mucosal adhesive aqueous gel applied in a single application contains an MTZ of about 60 mg to about 100 mg.

U4. The use according to any one of aspects U1 to U3, wherein the amount of mucosal adhesive aqueous gel applied in a single application contains approximately 65 mg of MTZ.

U5. The use according to any one of aspects U1 to U4, wherein the mucosal adhesive aqueous gel is applied intravaginally once daily for a period of 1 to 5 days.

U6. The use according to any one of embodiments U1 to U5, wherein the mucosal adhesive aqueous gel is applied intravaginally once daily for a period of one day.

U7. The use according to any one of aspects U1 to U5, wherein the mucosal adhesive aqueous gel is applied once daily for a period of 5 days. 6.5.4. Unit dose form

D1. A high dose suitable for vaginal application, comprising a certain amount of the gel according to any one of aspects C1 to C32 containing approximately 65 mg to approximately 100 mg of MTZ encapsulated in a container suitable for vaginal application. A unit dose form of mucosal adhesive MTZ aqueous gel.

D2. A unit dose form of embodiment D1, comprising an amount of gel containing approximately 65 mg of MTZ.

Unless otherwise defined, all technical and scientific terms used herein are common to those of ordinary skill in the art to which the various inventions described herein belong. Has the same meaning as what is understood. Methods and materials similar to or equivalent to those described herein may be used in the practice or testing of the disclosed invention, but suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, this specification, including the definitions, shall prevail. In addition, the materials, methods, and examples are merely exemplary and not limiting.

<p num="0218">7. Example Although the inventions of the present disclosure have been generally described herein, they are more easily understood by reference to the following examples provided as examples and do not limit the scope of the inventions described herein. Example 1: An exemplary solvent useful for solvent systems</p><p num="0219"> The solubility of MTZ in various solvents at 25 ° C was determined to identify solvents useful for dissolving MTZ at the concentrations required for the high dose mucosal adhesive MTZ aqueous gels described herein. did. The saturated MTZ solubility of many solvents at 25 ° C is shown in Table 1 below. Solvent systems useful for preparing high-dose mucosal adhesive MTZ aqueous gels with the desired concentration of MTZ can be devised using these saturated solubilities as a guide. Additional solvents suitable for use with these (and other) solvents for preparing the high dose mucosal adhesive MTZ aqueous gels described herein can be readily identified based on their saturated MTZ solubility. Table 2 below shows the saturated MTZ solubility at 25 ° C in a number of solvent systems mixed with an aqueous phase suitable for preparing high-dose mucosal adhesive MTZ aqueous gels.<tables num="1"><img id="000002" he="103" wi="148" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="2"><img id="000003" he="207" wi="69" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> Example 2: Illustrated high-dose mucosal adhesive MTZ aqueous gel</p><p num="0220"> A number of exemplary high-dose mucoadhesive MTZ aqueous gels containing different amounts of MTZ and utilizing a variety of different solvent systems were prepared and various different homogeneity tests, as described in the Examples below. It was subjected to rheology, stability, biological, and clinical trials. All gels were prepared using a similar process described below.</p><p num="0221"><u style="single">Gel preparation</u> The required amount of preservative (s), eg, methylparaben and propylparaben, were weighed and placed in a Duran® bottle, followed by the aqueous phase (phosphate buffer or deionized water). The inclusions were thoroughly mixed to dissolve the preservative. The remaining solvents (eg, benzyl alcohol, ethanol, propylene glycol, and PEG 400) were weighed and placed in separate Duran® bottles, mixed thoroughly, and then the required amount of metronidazole was added. The Duran® bottle was then placed in a 55 ° C water bath and stirred continuously until metronidazole was dissolved. The Duran® bottle was removed from the water bath and placed at room temperature for continued mixing until the solution equilibrated to room temperature. The storage solution was then added to the MTZ solution with constant stirring. The required amount of gelling agent was weighed and placed in a weighing boat, the solution was added with constant stirring and stirred until the gelling agent was completely hydrated. Care was taken to mix the inclusions thoroughly to ensure homogeneity.</p><p num="0222"> For gels that utilize pH-sensitive or pH-dependent gelling polymers, the pH of the MTZ solution can be adjusted to a value within the optimum range for gelation prior to the addition of the gelling polymer, and the pH of the gel will then be adjusted. Adjusted as needed.</p><p num="0223"><u style="single">Placebo gel preparation</u> In some of the experiments discussed herein, placebo gels are used as comparisons or controls. Bracebogel was prepared as described above. For placebo, MTZ was omitted and replaced with equal weight% water or buffer, depending on the aqueous phase used in the active gel.</p><p num="0224"><u style="single">Preparation of phosphate buffer pH 4</u> Disodium hydrogen phosphate dihydrate (3.9 g) is weighed and placed in a 500 mL Duran® bottle and 450 mL of deionized water is added. The solution is mixed thoroughly to dissolve the buffer salt. The pH of the solution was adjusted to pH 4 using o-phosphoric acid to a volume in a 500 mL volumetric flask with deionized water.</p><p num="0225"> The compositions of various exemplary gels containing 1.5% MTZ, 2.0% by weight, 1.3% by weight, and 1.18% by weight MTZ are shown in Tables 3, 4, 5, and 6 below, respectively.</p><p num="0226"> The formulations MG03, MG04, MG08, and MG09 listed in Table 3 formed a clear, highly viscous pale yellow gel, whereas MG26 formed a milky white gel. The ideal pH of carbomer, polycarbophil AA-1, and carbomer 974P for forming gels is neutral. Since the desired pH of the gel for vaginal application is about pH 4, large amounts of these gelling agents were used in comparison to HEC. MG03 contained β-hydroxypropyl cyclodextrin as a solubilizer.<tables num="3"><img id="000004" he="172" wi="85" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="4"><img id="000005" he="168" wi="90" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="5"><img id="000006" he="214" wi="88" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="6"><img id="000007" he="80" wi="155" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> Example 3: Mucosal adhesion properties of the exemplary gel</p><p num="0227"> Two exemplary high-dose mucosal adhesive MTZ gels (MG32PB and MG33PB) used two different methods to adhere to the isolated porcine gastric mucosa and leologic measurements with and without mucin. And tested. The method and results are described below. Mucosal adhesion using isolated porcine gastric mucosa</p><p num="0228"><u style="single">Method</u> For the experiment, a small piece of porcine gastric mucosa (15 x 10 cm) was placed on an incline at a 10 ° angle and washed with a peristaltic pump in pH 4 buffer for 10 minutes. A small aliquot (50 μL) test gel was spread on the mucosa on a slope and left for 10 minutes. The pump was then switched on and the buffer recovered after t = 1, 2, 3, 4, 5, 10, and 15 minutes was placed in separate vials at each time point. Blank samples without any test gel were also analyzed for any background or interference. Samples were filtered through a 0.45 μm PVDF syringe filter and analyzed by HPLC. The rate of loss of MTZ from the mucosa was plotted against time for each test gel and the data obtained were compared to determine relative mucosal adhesion of each test gel. A second experiment was also performed for 5 minutes at the time of sampling at 15 second intervals.</p><p num="0229"><u style="single">result</u> The data are shown in Figures 1 and 2. The results show that MG32PB has slightly better mucosal adhesion than MG33PB, as determined by the amount of metronidazole remaining on the mucosa at each time point. b. Rheology measurement</p><p num="0230"><u style="single">Method</u> Mucin was weighed and placed in a glass vial, followed by the gel under investigation. A magnetic follower was placed in the sample and mixed until all mucin was hydrated. Rheological parameters of the test gel were analyzed using vibration experiments with and without mucin (Bohlin CVO rheometer). An amplification sweep using the corresponding mucin-free placebo gel followed by a frequency sweep was performed first to determine the linear viscoelastic region. The parameters used to determine the frequency sweep of the gel are shown in Table 7.<tables num="7"><img id="000008" he="57" wi="134" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0231"> The test gel (approximately 1 g) was placed on the bottom plate of the rheometer fitted to the parallel plate. The lower plate was raised to capture the sample in a predetermined gap. The sample was then subjected to a series of sinusoidal vibrations from 5 to 20 Hz in 0.15 Hz increments. The applied torque (4Pa) was pre-determined to be within the linear viscoelastic region for both the highest and lowest frequencies. Each gel was tested 3 times. At the end of each run, Bohlin computer software (version 6.50.5.7) was used to determine the storage modulus G'. The data obtained for each parameter was plotted as a function of frequency and the average value of G'was calculated at a fixed frequency of 10 Hz. The log ratio of ΔG'to mucin G'with and without mucin using the average G'value of the mucin-containing active gel and compared to the average G'value of the mucin-free active gel. Was calculated.</p><p num="0232"><u style="single">result</u> The addition of mucin, which is also a polymer, increased the modulus of elasticity of the test gel as determined by ΔG'(Table 8). Higher log G'ratio values are expected to correlate with more mucin interactions and thus better mucosal adhesion. However, this increase may depend on the endogenous interaction between the gelled polymer used in the preparation of the gel and the mucin. The log G'ratio, which is the ratio of log G'of the test gel with and without mucin, provides insight into the increased viscoelastic properties of the formulation.<tables num="8"><img id="000009" he="33" wi="133" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> Example 4: Short-term storage stability of the exemplary gel</p><p num="0233"> Various different embodiments of the exemplary high-dose mucosal adhesive MTZ aqueous gels were tested for short-term (4 week) storage stability at 25 ° C and 45 ° C. The gel was placed in a closed amber vial and stored in a Vindon Scientific stability cabinet. The parameters tested included homogeneity (visual and microscopic observation of the presence of crystals and / or particles), MTZ stability, storage potency, viscosity, and pH. This stability was tested at 25 ± 2 ° C, 60 ± 5% RH and 40 ± 2 ° C, 75 ± 5% RH at t = 0, 2, and 4 weeks. Placebo gels without MTZ were also stored for the same period and evaluated for physical and chemical stability.</p><p num="0234"> To evaluate MTZ stability, MTZ was extracted from the gel and analyzed by HPLC method 1 (parameters in Table 9 below). For extraction, about 0.5 g gel was placed in a 50 mL volumetric flask. Approximately 30 mL of HPLC mobile phase 65:35 methanol / 60 mM phosphate buffer (prepared as follows) was added and the mixture was swirled until gel was found to be dispersed in the solution. The solution was volumetricized with an HPLC mobile phase. For HPLC analysis, the aliquot was filtered through a 0.45 μm PTFE syringe filter and the first 2 mL was discarded. The residual filtrate was analyzed by HPLC method 1.<tables num="9"><img id="000010" he="94" wi="133" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0235"> To prepare the mobile phase, 1.5 g of monobasic sodium phosphate and 1.3 g of dibasic sodium phosphate were weighed and placed in a Duran® bottle and dissolved in 350 mL of deionized water. The solutions were mixed thoroughly, 650 mL of methanol was added, the resulting solution was thoroughly mixed with a magnetic stirrer and filtered through a 0.22 μm filter.</p><p num="0236"> For HPLC analysis, MTZ peaks (retention time of about 3.312 minutes for 254 nm detection) were integrated and compared to control samples. The amount of MTZ remaining after storage was quantified as a percentage of theoretical concentration (TC) at t = 0. The measured concentration (MC) was determined using the calibration curve obtained with a known concentration of MTZ stock solution (prepared by serial dilution of 1 mg / mL stock).</p><p num="0237"> Physical stability of the gel was evaluated using a light microscope (Leica DME SOP) 3091). The test gel was compared to the corresponding placebo control to assess the presence of particles.</p><p num="0238"><u style="single">result</u> The percentage of MTZ recovered from the gel is shown in Table 10 below. MTZ was stable at all concentrations tested and in all gels. Low recovery was observed in MG37PB, MG42PB, and MG43PB from the sample at 25 ° C after 4 weeks. However, the recovery from the same gel stored at 40 ° C was 100%, suggesting a potential extraction / measurement error for the sample at 25 ° C. The peak purity% of all samples was 100%.<tables num="10"><img id="000011" he="98" wi="142" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> Example 5: Long-term storage stability of the exemplary gel</p><p num="0239"> Various different embodiments of the exemplary high-dose mucosal adhesive MTZ aqueous gels were tested for long-term stability at 25 ° C and 40 ° C.</p><p num="0240"> In this study, 20 g of each test gel was placed in a sealed vial at 25 ± 2 ° C / 60 ± 5%. Stored at RH and 40 ± 2 ° C / 75 ± 5% RH. Backup samples were also stored at -20 ° C, 2-8 ° C, and 30 ° C / 65% RH for analysis if either the real-time or acceleration-time samples failed. The corresponding placebo gel without MTZ was stored with the test gel.</p><p num="0241"> After 1, 2, 3, 6, 12, and 18 months, the gel was removed from the storage cabinet and tested for physical and chemical stability using microscopic and viscosity analysis as described below. ..</p><p num="0242"><u style="single">Microscopic analysis</u> The gel was viewed under a light microscope (Leica® DME). A small amount of gel sample was placed on a microscope slide using a microspatula. The microscope slide was covered with a cover glass and the gel was viewed using a 40x objective. Active gels were compared to placebo gels for the presence of crystals.</p><p num="0243"> If crystals were found, the particle size was measured using a scale on a calibrated Graticle (Olympus® Objective Micrometer 0.01 mm). The microscope was set up so that the camera (Nikon Coolpix® 4500 digital camera) could be attached to the relay lens of the microscope, and the 40x objective lens was set where the sample could be seen. Camera settings: Image size: 1280 x 960 pixels, Image quality: Fine. I took a picture (400x magnification) when I got a clear and clear view.</p><p num="0244"><u style="single">Viscosity analysis</u> Gel rheology was measured at each time point (t = 0, 1, 2, 3, 6, 12, and 18 months). A cone and plate measurement system for flow curve measurement is preferred because the shear rate is constant across the diameter of the cut cone. Therefore, a controlled shear rate ramp method was used. Shear velocity 0.16s^-1Each gel was compared and the viscosity was determined using points on the flow curve at. The rheometer settings used are shown in Table 11 below.<tables num="11"><img id="000012" he="57" wi="133" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0245"> Gels were also tested for MTZ content, benzyl alcohol content, paraben content, storage potency, and pH. With the exception of 18-month samples, MTZ content was evaluated as described above for short-term stable samples. For 18-month samples, the MTZ content was evaluated as described below.</p><p num="0246"> For extraction, 0.5 g ± 20 mg gel was weighed and placed in a 25 mL volumetric flask and diluted with approximately 20 mL sample diluent (3: 7 methanol / 10 mM KH).₂PO₄). The mixture was swirled until the gel was dispersed in the solution (about 5 minutes). The solution was volumetricized with sample diluent and vortex mixed for about 1 minute. Approximately 14 mL of sample was centrifuged at 3500 rpm for 10 minutes and the resulting supernatant was filtered through a 0.45 μm PTFE syringe filter and the first 2 mL was discarded. The residual filtrate was analyzed by HPLC method 2 (parameters are shown in Table 12 below). In addition to MTZ, recovery of benzyl alcohol, methylparaben, and propylparaben was also determined.<tables num="12"><img id="000013" he="126" wi="138" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0247"> Preservative efficacy tests were performed in a manner that complies with European and US Pharmacopeia specifications. Inoculated samples were tested on t = 0 hours, 24 hours, 48 hours, 7, 14, 21, and 28 days.</p><p num="0248"> Gels corresponding to MG33PB but containing reduced preservatives were tested for preservative efficacy. The test gels in Table 13 below were prepared as described in Example 2.<tables num="13"><img id="000014" he="54" wi="113" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0249"> Placebos for gels MG33 (75%) and MG33 (90%) were also prepared.</p><p num="0250"> pH tests were performed at t = 0, 1, 2, 3, 6, 12, and 18 months.</p><p num="0251"><u style="single">result</u> Table 14 shows the recovery rate of MTZ from the gel. The recovery of MTZ from gels stored at 25 ° C and 40 ° C for 6 months corresponds to t = 0 data. The peak purity% of all gels was found to be 100%, indicating that MTZ is stable in all gels tested. The recovery of MTZ from MG33PB and MG32PB stored at 25 ° C and 40 ° C for 18 months corresponds to t = 0 data.<tables num="14"><img id="000015" he="170" wi="101" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0252"> Physical stability of metronidazole gel formulation. Gels stored at 25 ° C and 40 ° C were visually and microscopically evaluated for the presence of any particles or crystals after 6 months. The gels were found to be physically stable, but there was a slight change in color from ultra-light yellow to light yellow in all active gels after 6 months storage at 40 ° C. This was not seen with the corresponding placebo formulation (Table 17). A slight difference in pH was observed during storage. However, no significant pH shift was observed in the active gel, indicating that the gel was stable after 6 months (Table 15).</p><p num="0253"> The viscosity of the gel was measured using a flow curve. The point of the flow curve comparing the viscosity of the gel is 0.16 s in shear rate.^-1(Table 11). After reading (n = 2) at t = 0, the curves were tested and points to compare were selected. All gels exhibited pseudo-plastic action or shear thinning, i.e., the viscosity decreased as the shear rate increased. As the shear rate increased, the instantaneous viscosities of the gels gradually resembled each other so that there was no significant difference between the viscosities of the gels. Points on the curve were selected for comparison between gels with all different flow curves. 0.16s^-1Viscosities were calculated using the interpolation of the two closest shear rates above and below. All gels after 6 months storage at 25 ° C, and MG33PB stored at 25 ° C for up to 18 months, showed slight variability in viscosity and were generally constant between each time point up to 18 months. It was found that there was (Table 16). A decrease in viscosity was observed after storage at 40 ° C, which was more pronounced for placebo gels than for active gels.</p><p num="0254"> Backup active gels stored at 2-8 ° C for 6 months were also tested for crystal formation, similar to placebo gels stored under the same conditions as the corresponding optimal gels. Large crystals were observed in gels MG32PB and MG33PB. Large sheet crystals were also formed on the gels MG33PB and MG36PB. Single small crystals formed in gels MG34PB and MG37PB. These data indicate that storage at 2-8 ° C is not recommended for high dose mucosal adhesive MTZ aqueous gels. Storage at these temperatures is more appropriate as no crystal formation was observed at 25 ° C or 40 ° C.</p><p num="0255"> Table 17 below shows observations of the physical appearance of placebo and the test product stored at 25 ° C and 40 ° C for 6 months.<tables num="15"><img id="000016" he="218" wi="107" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="16"><img id="000017" he="220" wi="126" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="17-1"><img id="000018" he="228" wi="126" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="17-2"><img id="000019" he="225" wi="76" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0256"><u style="single">PET test results of MG33PB stored at 40 ° C for 12 months</u> The results of the storage efficacy test (PET) are summarized in Tables 18 and 19. Gel MG33PB stored at 40 ° C for 12 months is consistent with the log reduction required for all organisms tested, as specified by European Pharmacopoeia 6.3.<tables num="18"><img id="000020" he="61" wi="143" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="19"><img id="000021" he="186" wi="56" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0257"><u style="single">Analysis of 18-month samples stored for 12 months using valid analysis method 2 and then stored at room temperature for an additional 6 months</u> Samples stored at 25 ° C for 12 months and then stored at ambient temperature for an additional 6 months, samples stored at 25 ° C and 40 ° C for 18 months meet MTZ and preservative content specifications. (Table 20 and Table 21).<tables num="20"><img id="000022" he="197" wi="56" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="21"><img id="000023" he="202" wi="62" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0258"><u style="single">Effect of 75 and 90% w / w preservative compositions on MG33PB</u> Tables 22 and 23 summarize the results of PET studies performed with variants of the exemplary gel MG33PB containing reduced doses of preservatives, MG33 (75%) and MG33 (90%). MG33 (75%) and MG33 (90%) meet the log reduction required for all organisms tested, as specified by the European Pharmacopoeia 6.3.<tables num="22"><img id="000024" he="52" wi="148" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="23"><img id="000025" he="199" wi="150" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables> Example 6: Topical delivery and skin permeation properties of the exemplary gel</p><p num="0259"> The in vitro skin permeation properties of some of the illustrated highly mucosally adherent MTZ aqueous gels were tested in Franzsel using the full layer of human abdominal skin formed from informed consent. For this study, all subcutaneous fat was removed with a scalpel and the skin was placed between the donating and receiving compartments.</p><p num="0260"> For this experiment¹⁴C Radiation-labeled MTZ gels were prepared as described in Example 2 with the amounts of components listed in Table 24 below. In Table 24, all amounts are mg¹⁴The exception is the C-labeled MTZ, which is μL.<tables num="24"><img id="000026" he="200" wi="105" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0261">¹⁴The C radiolabeled MTZ (GE Healthcare) had a specific activity of 57 mCi / mmol with a radiochemical purity of 99.2%.¹⁴C-labeled MTZ stock solution is 37 MBq of 3.5 mL of water¹⁴Add to C-labeled MTZ,¹⁴Prepared by vortex mixing until the C-labeled MTZ was dissolved.</p><p num="0262"> As a control, 10g 0.75% by weight corresponding to FDA approved 0.75% by weight MTZ gel (eg METROGEL VAGINAL®)¹⁴A C-labeled MTZ gel was prepared as follows.</p><p num="0263"><u style="single">Paraben phase</u>: Methylparaben (8.03 mg), propylparaben (2.05 mg), and propylene glycol (303.28 mg) were weighed and placed in a 28 mL glass vial and stirred until dissolved. The vial was then equilibrated at 55 ° C and water (3.5 g) was added with stirring. MTZ (75 mg) was added and stirring was continued until all drugs were dissolved. next¹⁴A C-labeled MTZ stock solution (465 μL) was added with stirring.</p><p num="0264"><u style="single">Carbomer phase</u>: Disodium EDTA (5.03 mg) was weighed and placed in a 28 mL glass vial bottle supplemented with purified water (5.698 g). Using magnetic furia, the formulation was stirred until all disodium EDTA was completely dissolved. Carbomer 974P (200.89 mg) was added and the formulation was stirred overnight to hydrate the polymer.</p><p num="0265"><u style="single">composite</u>: The paraben phase was added to the carbomer phase with stirring. Water (200 mg) was used to pour the paraben phase into the carbomer phase. The gel was left overnight to cool to room temperature.</p><p num="0266"> For this experiment, full-thickness human skin was placed in Franz cell with pH 4 phosphate buffer (pH 4) as a receptor to ensure sedimentation conditions. 10mg / cm²Apply a finite amount of test gel equivalent to¹⁴The diffusion of C-labeled MTZ was determined over time.</p><p num="0267"> A test gel (7 μL) was applied to the surface of the membrane using a positive displacement pipette. Since many gels were tested, the study was divided into several separate experiments. Two skin donors were randomly assigned over all experiments so that each gel was tested on both skin donors (n = 6 cells / gel).</p><p num="0268"> The receiving compartment of Franz cell was filled with receptive fluid and the cell was fixed in a water bath maintained at 37 ° C. The contents of the receiving chamber were continuously stirred by a small magnetic follower. Samples of receptive fluid were taken from the receptive compartment at t = 1, 2, 3, 4, 6, 8 and 24 hours, replaced with fresh receptive medium and analyzed by scintillation counting.</p><p num="0269"> At the end of the experiment, mass balance was performed and the donor chamber, surface residue, stratum corneum (SC), residual epithelium, dermis, and receiving chamber were analyzed. This method involved removal of SC by tape stripping, as well as treatment of the residual epithelial layer and dermis using standard procedures. This method is briefly described below.</p><p num="0270"><u style="single">Non-absorbable product</u>: The surface of each Franz cell donor chamber was gently wiped with a cotton swab 5 times clockwise and 5 times counterclockwise. This procedure was repeated 4 times, alternating between wet and dry swabs (pre-soaked in the receptive solution). A cotton swab was added to the scintillation cocktail prior to analysis. Two pieces of tape were removed from the skin and considered as non-absorbent and included in the total surface activity. The tape pieces were placed in scintillation vials with 1 mL of water added. These were immersed for 72 hours to dissolve the pharmaceuticals and disperse them in water. Prior to analysis by liquid scintillation (LSC), a scintillation cocktail (4 mL) was added to the vial. The surface of each Franz cell receiving chamber was gently wiped with a cotton swab 5 times clockwise and 5 times counterclockwise. This procedure was repeated twice, alternating between wet and dry swabs (pre-soaked in the receptive solution). A cotton swab was added to the scintillation cocktail prior to analysis.</p><p num="0271"><u style="single">Stratum corneum (SC)</u>: SC was removed by carefully tape-peeling the skin 10 times using Scotch adhesive tape. The first five pieces of tape were placed together in one scintillation vial and the next five pieces were placed together in a second vial. 1 mL of water was placed in each vial and these were soaked for 72 hours to dissolve the formulation and disperse it in water. Prior to analysis by LSC, a scintillation cocktail (4 mL) was added to the vial.</p><p num="0272"><u style="single">Epithelium</u>: Residual epithelial fragments (after tape removal) were carefully removed from the dermis with a scalpel. The epithelium was placed in a glass vial containing 2 mL Soluene 350 and cultured at 50 ° C for 72 hours prior to analysis by LSC.</p><p num="0273"><u style="single">Dermis</u>: The residual dermis layer was placed in a glass vial containing 2 mL Soluene 350 and cultured at 50 ° C for 72 hours prior to analysis by LSC.</p><p num="0274"><u style="single">result</u> Each gel tested was recovered from different skin layers¹⁴The amount of C-labeled MTZ is shown in Table 25 below. These are schematized in Figure 3A. The same data, represented as the percentage of dose applied, is shown in Table 26 below and schematized in Figure 4A.</p><p num="0275"> Additional diagrams are provided in Figures 5A and 6A.</p><p num="0276"> There were no statistical differences in the amount of MTZ detected in the donor chamber, receiving chamber, epithelium, and dermis for any of the formulations tested (p&gt; 0.05). However, when using a high-dose MTZ gel, there was a significant difference in the amount detected in the receptor and stratum corneum compared to 0.75 wt% Metrogel®. The level of MTZ observed in the receptor after application of Metrogel® (4.83 ± 0.97 μg) is as low as 0.31 ± 0.27 from the new PB formulation (MG33PB to 0.31 ± 0.27) with the possible exception of MG37PB (3.62 ± 2.02 μg). It was 4 to 16 times higher than μg (up to 1.17 ± 0.25 μg from MG36PB) (2 to 9 times when normalized for concentration; p &lt;0.05). However, in the stratum corneum after application of the exemplary test gel¹⁴C-labeled MTZ levels (minimum 17.80 ± 3.45 μg from MG36PB, maximum 33.15 ± 19.86 μg from MG35PB) were 50-90 times higher than those obtained for Metrogel® (0.37 ± 0.74 μg) ( 29-53 times when normalized to concentration). This difference is reflected in the amount of non-absorbable radiolabeled MTZ recovered from the skin surface, and the amount of drug recovered from the Metrogel® formulation (45.03 ± 1.20 μg) is due to the difference in initial concentration. Nevertheless, it corresponded to the exemplary test gel (minimum 55.69 ± 5.42 μg from MG33PB, maximum 65.23 ± 3.29 μg from MG34PB).</p><p num="0277"> Similar experiments were performed with an exemplary gel containing 1.5 wt% MTZ and similar results were obtained (provided in Figures 3B, 4B, 5B, and 6B).</p><p num="0278"> Therefore, compared to the conventional 0.75 wt% MTZ, the high-dose mucosal adhesive MTZ aqueous gels described herein retain large amounts of MTZ on the surface of the skin, while being lower in the receptive fluid. It produces levels and corresponds to the low systemic levels expected when applied topically to the skin.<tables num="25"><img id="000027" he="222" wi="58" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="26"><img id="000028" he="222" wi="59" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables>Example 7: Permeation study using porcine vaginal tissue</p><p num="0279"> Permeation studies using the exemplary test gel MG33PB and a 0.75 wt% MTZ control gel similar to that described in Example 6 were performed in Franz cell using porcine vaginal tissue from which excess muscle had been removed. It was. MTZ was extracted from the tissue using Soluene. In this experiment, the gel¹⁴Did not contain C-labeled MTZ. MTZ recovery was measured via HPLC. The average amount of MTZ recovered from the vaginal tissue surface and within the vaginal tissue 24 hours after application of the test gel or control gel is shown in Table 27 below. In the data below, n = 5, and one outlier was excluded from each of the test gel and control experiments.<tables num="27"><img id="000029" he="32" wi="152" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0280"> The data is schematized in FIG. The cumulative amount of MTZ that permeated vaginal tissue as a function of time throughout the course of the experiment is shown in Figure 8. In FIG. 8, the data are vaginal tissue (μg / cm).²) Is expressed as the average (± standard error) amount (μg) of the transmitted MTZ per unit area. Both test and control batches have N = 5.</p><p num="0281"> In this experiment, only about 30% of the total amount of MTZ applied to both the test gel and the control gel was recovered.</p><p num="0282"> Only about 30% of the total amount of MTZ was recovered from the study and control groups in this experiment. Example 8: An exemplary high-dose mucosal water-based gel containing 1.3 wt% MTZ is surprisingly effective and FDA-approved when used clinically to treat women diagnosed with bacterial vaginosis. Much more effective than 0.75 wt% MTZ gel</p><p num="0283"><u style="single">Introduction</u> A clinical evaluation of a specific embodiment of a high-dose mucosal adhesive MTZ aqueous gel containing 1.3 wt% MTZ and the preparation MG33PB was performed, and its safety and efficacy were evaluated. In this study, MG33PB was evaluated against 0.75 wt% METROGEL VAGINAL® ("MGV") and different study groups were used to determine efficacy at days 1, 3, and 5.</p><p num="0284"><u style="single">General method</u> Multicenter, randomized, inspector-blind, administration of MG33PB (QD x 1 day, QD x 3 consecutive days, and QD x 5 consecutive days) compared to MGV (QD x 5 consecutive days) for the treatment of BV Quantitative efficacy and safety studies were conducted in accordance with FDA guidance (Draft FDA Guidance dated July 1998, entitled "Guidance for Industry: Bacterial Vaginosis-Developing Antimicrobial Drugs for Treatment"). Subjects were evaluated at three time points: screening / baseline visits, post-treatment phone calls, and end-of-study [EOS] / cure study [TOC] visits. Subjects participated for up to 30 days. The study design is outlined in Table 28.<tables num="28"><img id="000030" he="26" wi="148" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0285"><u style="single">Research group</u> Participants who met all of the inclusion criteria listed below were allowed to participate in the study. Those who met one of the exclusion criteria listed below were not admitted to the study.<u style="single">Inclusion criteria</u> 1. Can voluntarily give informed consent in writing. 2. A woman at least 18 years old. 3. Good general health. There is no known medical history that the inspector determines may prevent participation in the study, as confirmed by medical history at the time of the screening visit. 4. Be able to voluntarily participate in the study as an outpatient, visit the research center when needed, and comply with all research requirements, including concomitant medications and other treatment restrictions. 5. For women of childbearing potential, urine pregnancy tests are negative before the start of study treatment. 6. Have a clinical diagnosis of BV, defined by having all of the following criteria: 1. Off-white (milky or gray) thin homogeneous secretions 2. Presence of glomerular cells in excess of 20% of total epithelial cells in saline wet mount microscopic testing Vaginal fluid pH above 3.4.7 4.10% KOH odor test positive 7. Agree to refrain from sexual intercourse for the first 7 days of the study. After the first 7 days, agree to use non-lubricated condoms when having sexual intercourse. 8. Accept to refrain from alcohol inoculation during and one day after treatment. 9. Agree to avoid the use of vaginal products during the study period (eg bidets, women's odor sprays, spermicides, lubricating condoms, tampons, and pessaries).<u style="single"> Exclusion criteria</u> 1. You plan to become pregnant, breastfeeding, or during the study period. 2. Menstruating at the time of diagnosis or expected to begin during the treatment period of the study. 3. Experience clinically significant medical events within 90 days of the visit (eg, stroke, myocardial infarction, etc.). 4. Any other known or suspected cause of infection of vaginitis (eg, candidiasis, trichomonas vaginalis, chlamydia trachomatis, gonorrhea, or active simple herpes) or, at the discretion of the inspector, confusing the interpretation of the clinical response. Has other vaginal or vaginal conditions (Chlamydia trachomatis and gonococcal test results are not available at the time of randomization). It has a Gram-stained slide Newgent score of less than 5.4 (results are not available at the time of randomization, but are used to determine the post-study analytical population). 6. Received antifungal or antibacterial treatment (systemic or intravaginal) within 14 days of randomization. Antiviral treatment (non-vaginal) is acceptable. 7. Disulfiram was inoculated within 14 days of randomization. 8. Pre-hypersensitivity to metronidazole given orally or topically or in any form of paraben. 9. Have primary or secondary immunodeficiency. 10. During the study period, cervical intraepithelial neoplasia (CIN) or cervical cancer is being treated or will be treated. 11. I am using anticoagulant therapy with Kumazine (Wafarin). 12. You are using systemic corticosteroids or systemic antibiotics. 13. Have previously enrolled in this study. 14. Participated in another clinical trial or ingested an experimental drug or device within the last 30 days. 15. Does not or cannot comply with protocol requirements.</p><p num="0286"><u style="single">Treatment group</u> After all screening / baseline assessments and assessments have been completed, the subject is instructed on appropriate techniques for the application of the study drug, and an open-label drug dosing coordinator is assigned to one of the following treatment groups 1: 1: 1: 1: Randomly assigned in a ratio of 1. . MGV: QD x 5 consecutive days . MG33PB: QD x 1 day . MG33PB: QD x 3 consecutive days, and . MG33PB: QD x 5 consecutive days</p><p num="0287"> Each subject received one kit of study drug. Each kit contains one of the following: -Research drug, MG33PB, or MG33PB, supplied with the appropriate number of applicators in a 70 g tube. -MGV, a comparative drug supplied by 5 applicators in a 70 g tube.</p><p num="0288"> Outcomes of various subgroup populations were evaluated (more detailed below). "<u style="single">Comprehensive analysis</u>The (ITT) population includes all randomized subjects in the study. "<u style="single">Modified Comprehensive Analysis "</u>The (MITT) population included all randomized subjects who received any study drug, underwent at least one post-treatment baseline visit / assessment, received a negative test for N. gonorrhoeae and Chlamydia trachomatis, and visited. It had a Gram-stained slide nougant score of 4 or higher at 1. Subjects lacking primary efficacy data were counted as failed primary efficacy endpoints and included in the MITT population. "<u style="single">Par protocol</u>The (PP) population included subjects from the MIT population that met the following criteria: -Meets all inclusion and exclusion criteria and has no protocol violations. Start the study drug on the day of randomization or within 2 days of the date of randomization. . Comply with research drugs. Do not use antibiotics during the study period (from randomization to TOC visits) (other than the permissible par protocol). Do not use additional vaginal products during the study period. -TOC Gram stain Nugent score results were obtained between 20 and 31 days relative to the first day of treatment. Data show that subjects whose study visits ended prior to day 21 had a clinical failure of another unspecified cause of bacterial vaginosis (ie, Trichomonas, Chlamydia, gonorrhea). If so, it was included in the PP group.</p><p num="0289"> 「<u style="single">safety</u>The population included all randomized subjects to whom any amount of study drug was applied.</p><p num="0290"><u style="single">Blinded</u> Because the study and comparative drugs were administered using different treatment regimens, the inspector-blind study design was utilized. Treatment assignments were not known to the inspectors and research coordinators at each institution where the treatment assignments and medications were given, as well as the independent dosing coordinators.</p><p num="0291"><u style="single">Limited medication / treatment</u>The following drugs, preparations, and treatments that may affect the results of the study were prohibited during this study. 1. Alcohol intake during and 1 day after treatment 2. Disulfiram during and 1 day after treatment 3. Vaginal products during the study period (eg bidets, women's deodorant sprays, spermicides, lubricating condoms, tampons, and pessaries) 4. Systemic or vaginal antibacterial therapy during the study period, with the exception of oral antifungal therapy (eg, oral fluconazole) to treat comorbidities (eg, Candida). Antiviral therapy (non-vaginal) was tolerated. 5. Treatment of CIN or cervical cancer 6. Kumajin or Warfarin 7. Systemic corticosteroids</p><p num="0292"><u style="single">Effectiveness and safety variables</u> Efficacy analysis was performed on both PP and MITT populations.</p><p num="0293"> The primary efficacy variable will be evaluated at Visit 2 (EOS / TOC) as evidence of therapeutic cure. Subjects considered to be therapeutically cured must have achieved both clinical and bacteriological cures at EOS / TOC.</p><p num="0294"> Clinical cure is defined as a solution to the clinical findings obtained from the baseline visit and is further defined as: 1. Absence of off-white (milky or gray) thin homogeneous secretions 2.10% KOH odor test negative 3. Absence of filamentous cells in saline wet mount 4. Vaginal fluid pH less than 4.7</p><p num="0295"> In addition, the subject must not have received any antimicrobial agents (other than the permissible perprotocol) during the study period, and the inspector said at the time of EOS, "In your opinion, the patient is infected with BV at this point. You must answer "no" to the question "Do you need additional treatment?"</p><p num="0296"> Bacteriological cure is defined as a Nugent score of less than 4.</p><p num="0297"> The key secondary effectiveness variables for this study are: . Percentage of subjects with clinical cure at TOC / EOS visit Percentage of subjects with bacteriological cure at TOC / EOS visit The time it takes to resolve symptoms (abnormal secretions and odors), defined as the time interval (days) from randomization to the day the subject's diary indicates resolution of symptoms. Pelvic test results including pruritus, irritation, and inflammation . Target questionnaire and diary data</p><p num="0298"> Subjects classified as clinical failures on unscheduled interim visits advanced their clinical and bacteriological outcomes and treatment assessments to the EOS / TOC visit.</p><p num="0299"> A safety analysis was performed on the safety population and included an evaluation of vaginitis history and previous treatment, pelvic studies, concomitant medication monitoring, and AE / SAE monitoring.</p><p num="0300"><u style="single">Statistical methods and analysis</u></p><p num="0301"><u style="single">Analytical research means</u> The various research groups are defined above. The ITT population was used to summarize the subject's nature, demographic and basic characteristics, medical history, and previous drug / concomitant medications. The MITT population was used for symptomatic efficacy analysis. The PP population was used for the primary analysis. The safety population was used for study drug exposure and all safety analyzes. Subjects were analyzed as treated.</p><p num="0302"><u style="single">Statistical analysis of primary and secondary endpoints</u> For population variables, the data were organized by treatment group with an accurate 95% confidence interval (CI). Subjects who received other antibiotics during the study (ie, at any time from the date of screening / randomization to the date of the last visit), that is, anything other than the permissible perprotocol, were counted as failures.</p><p num="0303"> The Kaplan-Meyer survival curve for time to resolution was plotted for all treatment groups.</p><p num="0304"> No formal statistical trials were conducted to compare treatment groups. The final choice of treatment plan was based on cure rate, consistency of results, safety, and convenience information.</p><p num="0305"><u style="single">Safety analysis</u> Safety data are associated with all treatment-related adverse events (TEAEs), any serious adverse events (AEs), TEAEs, serious TEAEs, and AEs that result in study cessation for each treatment group. Consists of evaluations (number and%) of treatments to be performed.</p><p num="0306"> The inspector selects the following terms: mild, moderate, or severe to describe the severity of AE. The inspector evaluated the relationship between each AE and the study drug as unrelated, perhaps unrelated, perhaps related, or related. Note: Treatment-related AEs include those that are probably related or classified as related.</p><p num="0307"> Adverse events (TEAEs) that occur during treatment have an onset time at or after the start of the study drug and less than 30 days after the last dose of the study drug, or continue at the start of the study drug. It is defined as an adverse event that has become more severe or closely related to the study drug during treatment or during the 30-day period from the last dose of the study drug.</p><p num="0308"> The TEAE was summarized by the overall incidence of at least one event, the incidence by organ classification (SOC), and the incidence by SOC and preferred time period. TEAEs are also summarized by severity and association with research products. Adverse events that occurred during the following treatments were summarized as follows: severe TEAEs, treatment-related TEAEs, severe TEAEs and TEAEs leading to discontinuation of research.</p><p num="0309"><u style="single">Results of clinical evaluation of safety and efficacy</u></p><p num="0310"><u style="single">Target processing</u> The processing of the research subject is shown in Table 29 below. A total of 255 subjects were randomly assigned to 4 study treatments and were administered to all but 1 (MGV group) and included in the safety population. 234 subjects (91.8%) completed the study. Of the 21 subjects (8.2%) who discontinued early, 9 (3.5%) were unable to follow up and 5 (2.0%) were positive for gonococcal and / or Chlamydia trachomatis screening.</p><p num="0311"> The ITT population had 255 subjects (100.0%), the MIT population had 228 subjects (89.4%), and the PP population had 189 subjects (74.1%). Twenty-seven (10.6%) subjects were excluded from the MITT population, and the majority of 23 (90.0%) were due to baseline nougant scores. Sixty-six (25.9%) subjects were excluded from the PP population, and the most common reason that occurred in 30 (11.8%) subjects was a nugent score within 20-31 days of the first dose of the study drug. It wasn't. A summary of the target population is shown in Table 30 below.</p><p num="0312"><u style="single">Demographics</u> At baseline, the mean age of subjects enrolled in the study was 35.1 (± 9.93) years. Subjects were evenly divided into groups under 35 years (51.4%) and groups over 35 years (48.6%) and balanced by age range across the treatment group. A summary of demographics is shown in Table 31 below.<tables num="29"><img id="000031" he="223" wi="89" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="30"><img id="000032" he="221" wi="94" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="31"><img id="000033" he="222" wi="103" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0313"><u style="single">History of bacterial vaginosis and treatment of previous bacterial vaginosis</u> The history of bacterial vaginosis (BV) and previous treatments for BV are summarized in Table 32 below for the ITT population.<tables num="32"><img id="000034" he="90" wi="148" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0314"> Overall, 95 (37.3%) subjects reported previous episodes of BV, with an average duration of current BV episodes of 151.6 days. The treatment groups were similar with respect to the history and baseline characteristics of bacterial vaginosis.</p><p num="0315"><u style="single">Validity result</u> Summary of cure rate. A summary of cure rates in the PP and MITT populations (primary end point therapeutic cure rate and secondary end point clinical and bacteriological cure rate) is shown in Table 33 below.<tables num="33"><img id="000035" he="67" wi="145" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0316"> The results of the MITT population are similar to those of the PP population, as shown in Table 34 below.<tables num="34"><img id="000036" he="67" wi="145" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0317"><u style="single">Summary of presence / absence according to each therapeutic cure standard at the time of study completion / visit</u> For each of the four defined therapeutic cure criteria, the results of the presence or absence of EOS / TOC visits are presented in Table 35 for the PP population. The results are similar to the MITT population.<tables num="35"><img id="000037" he="129" wi="147" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0318"><u style="single">Time to resolve symptoms</u> In the PP population, the median time to resolve both abnormal secretions and fishy odors, as well as the median time to resolve fishy odors alone, was MGV 0. in all three metronidazole 1.3% treatment groups. Shorter than the 75% group (median time was 5 days vs. 6 days and 2 days vs. 3 days, respectively), and the median time to resolve abnormal secretions was equal in all treatment groups (median time) The median time is 3 days). Table 36 below summarizes the time it takes to resolve the symptoms (s).<tables num="36"><img id="000038" he="68" wi="147" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0319"><u style="single">Summary of therapeutic cure rates by subgroup</u> Therapeutic cure rates in the PP population are presented by subgroup in Table 37 below (age group, race, ethnicity, duration of current BV episode at baseline, and previous episodes of BV).<tables num="37"><img id="000039" he="114" wi="147" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0320"> Subgroup analysis showed no significant trend in achieving therapeutic cure.</p><p num="0321"><u style="single">Question result to the target</u> At the end of the study, we asked all subjects to fill out a questionnaire. Ask specific subjects the following questions and provide their feedback on the use of research drugs. How easy was it to apply the research drug? How convenient was the length of treatment with the research drug? How satisfied are you with the treatment you received? If you have BV symptoms in the future, do you prefer treatment with this study drug or would you like another treatment?</p><p num="0322"> A summary of the results is shown in Table 38 below.<tables num="38"><img id="000040" he="127" wi="149" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0323"> Of the four treatment groups, MG33PB was most likely to rate this treatment as "very easy to apply," "very convenient," (in terms of length of treatment), and "very satisfied." It was a 1-day group (data not shown).</p><p num="0324"><u style="single">Safety results</u> Drug exposure. In this 30-day study, subjects received MGV for QD x 5 consecutive days, MG33PB for QD x 1 day, MG33PB for QD x 3 consecutive days, or MG33PB for QD x 5 consecutive days. A summary of the mean days of drug exposure in the safety population study is shown in Table 39 below.<tables num="39"><img id="000041" he="33" wi="149" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0325"> In all treatment groups, the majority of subjects reported no or minimal leaks of study products.</p><p num="0326"><u style="single">Summary of adverse events</u> Overall, 92 (36.2%) subjects reported adverse events (AR). All adverse events were adverse events (TEAEs) that occurred during treatment. Twenty-nine (11.4%) subjects had TEAEs across the four treatment groups, and the examiner assessed them as related to the study drug. A summary of AEs is shown in Table 40 below.<u style="single">Adverse events that occurred during treatment</u> The most frequently reported organ-specific major categories (SOCs) across TEAE are infection and invasion (47 [18.5%] subjects), nervous system disorders (32 [12.6%] subjects), and reproductive and mammary gland disorders (subjects). 27 [10.6%] subjects). A summary of all TEAEs by Organ Classification (SOC) is shown in Table 41 below.<tables num="40"><img id="000042" he="219" wi="77" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="41"><img id="000043" he="222" wi="89" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0327"> The most frequently reported TEAEs with vulvar vaginal candidiasis (25 [9.8%] subjects) and headache (21 [21] 8.3%] Target). The table below summarizes all TEAEs with an incidence of&gt; 2.0% of the total subject, and those reported with an incidence of&gt; 2.0% of the total subject, regardless of their relationship to the study drug, in descending order. Shown in 42.</p><p num="0328"> Perhaps the most frequently reported TEAEs associated with or related to research drugs are vulvar vaginal candidiasis in the SOC category "Infection and Invasion" and "Reproductive and Breast Diseases" SOC. Vaginal itching and burning sensation. Table 43 below summarizes TEAEs in relation to SOC, preferred time period, and study drugs with an overall incidence of at least 6.0%.<tables num="42"><img id="000044" he="221" wi="57" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="43-1"><img id="000045" he="220" wi="71" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables><tables num="43-2"><img id="000046" he="221" wi="127" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0329"> The majority of adverse events were mild or moderate. A total of 5 subjects reported 6 severe events, including hypoglycemia, dizziness, abdominal pain, tooth abscess, vulvar edema, and burning sensation in the vagina. One subject (# 2904, 1.3% 1 day) reported two severe events that may be associated with the study product: vaginal burning sensation and vulvar edema that began on day 3 of the study. These events co-existed with pruritus vulva, headache, low back pain, and vulvar vaginal candidiasis. There were no differences between treatment groups in the incidence of severe AE.</p><p num="0330"><u style="single">Severe adverse events</u> During this study, 1 (1.5%) subjects had 1 SAE and hypoglycemia in the MG33PB QD x 1-day treatment group.</p><p num="0331"><u style="single">Summary</u> In this example, the efficacy and safety of 1.3% vaginal gel (test MG33PB gel) against bacterial vaginosis was explored. MG33PB once daily for 1, 3, or 5 days is similar to or numerically better therapeutic cure rate of over-the-counter MGV 0.75% vaginal gel (MGV) applied once daily for a total of 5 days. (Tables 33 to 36). Analysis of the subgroups analyzed showed no tendency for efficacy (Table 37). The highest percentage of subjects rated treatment as "very easy to apply," "very convenient in length of treatment," and "very satisfied" were in the MG33PB 1-day group (Table). 38). The majority of adverse events were found to be mild and moderate (Tables 40-43). Surprisingly, it was found that the 5-day treatment plan with MG33PB did not result in the observed post-treatment vulvar vaginal candidiasis (Table 42). Example 9: Antibacterial efficacy in an in vitro skin infection model</p><p num="0332"> The antibacterial efficacy of some exemplary high-dose MTZ gels against Gardnerella baginalis has been demonstrated in in vitro experiments. In this experiment, it was treated with a test gel and recovered from ATP from quantified infected skin. The lower the amount of ATP recovered, the more effective the gel is against infection.</p><p num="0333"> Two studies: Pilot studies and full-scale studies were conducted. The general method of pilot research is as follows. -Fresh topcoats were prepared according to standard protocols and cryopreserved at -20 ° C until needed. The membrane was cut into 3 mm x 3 mm sections and heat treated at 60 ° C for 15 minutes (previous experiments demonstrated that this was the optimum heating time to reduce background interference from the topcoat. . No data presented), 10 μL Gardonerella baginalis (approx. 5 × 10)⁷cfu) Infected using cell suspension. The viable cell count was measured to obtain the number of cell-forming units after the preparation of the cell suspension. -ChubTur® cells were set up and 3 mL of Ringer's solution was added to the cells to ensure a humidity-controlled environment. -After culturing at 35 ° C for 24 hours, 10 μL of test gel was applied on the surface of the topcoat. Following culture of the inoculated topcoat, dosing was performed by applying 2 μL of test gel to the topcoat sample (n = 3 for each gel and start time point, do not measure multiple time points from the same sample). ). Samples were analyzed for the presence of ATP from living organisms at 4, 8, and 24 hours following dosing. -Includes positive and negative controls. Negative controls include a topcoat sample with no organism or formulation added at t = 0, and positive controls include a topcoat sample with only an organism added (no formulation added) at t = 0. It is.</p><p num="0334"> For full-scale studies, the following changes were made to the protocol: -The initial infection of the epithelium and the anaerobic culture time before administration were reduced to 30 minutes. The gel exposure time was reduced to 2 hours to minimize the natural decrease in organism viability over time. -ChubTur® cells were removed from the culture and an ATP assay was performed to test the effectiveness of the gel 2 hours after application of the gel to a topcoat sample infected with Gardnerella baginalis.</p><p num="0335"><u style="single">Results: Pilot study</u> Figure 9 shows the variation in ATP release from an EP sample infected with Gardnerella baginalis (10 application administration) and its untreated infected EP sample when the exemplary active gel MG32PB (n = 3, at initiation) was applied. The comparison is shown. The corresponding placebo, MG32PB-P, and control were tested at n = 2. The lower the amount of ATP recovered, the more effective the test gel was for the test organism.</p><p num="0336"> The data presented in Figure 10 showed that positive controls (infected, untreated epithelium) resulted in high ATP recovery, as expected, while negative controls had low amounts of background interference. It was observed (uninfected, untreated epidermis). The administration time of the drug also shows the effect on ATP recovery. It can be seen that following 24-hour dosing, ATP levels decrease near background ATP (negative control) levels. This phenomenon was observed at 8 hours to varying degrees. From this pilot study, it was determined that the initial culture time (4, 8, and 24 hours) should be reduced to reduce the effect of spontaneous cell death over time. In a full-scale study, the dosing time was reduced to 2 hours.</p><p num="0337"><u style="single">Results: Full-scale study</u> Figure 11 shows the variation in ATP released from the epidermal sample infected with Gardnerella baginalis (ATCC14018) when the test gel was applied (n = 6 at the start of the active test gel and n = 3 for placebo). And a comparison with the over-the-counter comparative drug Metrogel®. Positive controls (untreated topcoat samples infected with Gardnerella baginaris) and negative controls (uninfected, untreated topcoat samples) were also included.</p><p num="0338"> The data presented in FIG. 11 show a reduction in ATP recovery in infected skin samples treated with the activity test gel when compared to recovery of ATP from the corresponding placebo gel-treated infected skin samples. Is shown. The product MG32PB showed a significant difference in efficacy between the active product and placebo with respect to the recovered ATP levels (data not shown). There is a significant difference (p &lt;0.05) showing a significant reduction in ATP recovery of the active formulation MG32PB compared to the infected subject, indicating the favorable efficacy of the formulation.</p><p num="0339"> Infected skin samples treated with the exemplary test gel MG33PB showed significantly lower (p &lt;0.05) ATP recovery for both the active formulation and placebo compared to the infected control. Considering the main differences between this formulation and other formulations, the efficacy of the formulation MG33PB placebo is expected because the polycarbofyl AA-1, bioadhesive polymer and excipients are present only in the MG33PB formulation. It was outside.</p><p num="0340"> The product MG35PB had little or no effect on reducing the viability of the organism for the active product when compared to infected samples that had not been previously treated with any of the products.</p><p num="0341"> All of the formulations MG32PB-A, MG33PB-A, and MG33PB-P showed comparable levels of activity against gardonerella baginaris on the human epithelium (P &lt;0.05). .. However, Metrogel® was significantly more effective when compared to MG32PB-P, MG33PB-A, and MG35PB-P (p &lt;0.05). ..</p><p num="0342"><u style="single">Summary</u> MG33PB-A and MG32PB-A were found to have higher efficacy than other formulations. Surprisingly, however, MG33PB-P (without metronidazole) was found to be as effective against Gardonerella baginalis as MG33PB (1.3% metronidazole). Example 10: In vitro evaluation of a novel metronidazole formulation for antibacterial efficacy using a disc diffusion inhibitory assay</p><p num="0343"><u style="single">Introduction</u> Studies were conducted to determine the minimum amount of benzyl alcohol required to suppress microbial growth. In this study, the antibacterial efficacy of three placebo and one active high-dose MTZ gel against Gardonerella baginaris was tested by a disk diffusion inhibitory assay.</p><p num="0344"> The gels tested are shown in Table 44 below.<tables num="44"><img id="000047" he="69" wi="149" file="JP6235087B2_D0001.tif" img-format="tif" img-content="drawing" /></tables></p><p num="0345"> Approximately 1 x 10 for the assay⁹A 3 mL Gardonerella baginalis suspension prepared at cfu / mL was prepared. A 100 μL aliquot suspension was pipetted and placed on the surface of a pre-injection Columbia blood agar (CBA) plate and the suspension was spread evenly over the surface of the agar medium using a sterile sprayer. The agar plate was then dried under a laminar flow cabinet. Inhibition assay was then performed by applying 20 μL of the test gel to the surface of the 1/4 antibiotic assay disc (n = 6 for each gel, note: disc contains no antibiotics) and under a laminar flow cabinet. Air dried for 1 minute. The discs were then inverted and Gardnerella baginaris was placed on the surface of a pre-sown plate and cultured at 35 ° C. for 48 hours in an anaerobic bottle. Anaerobic conditions were maintained by the use of AnaeroGen gas packs. Following culturing, the inhibitory areas of all formulations were measured with calipers.</p><p num="0346"><u style="single">result</u> The results shown in FIG. 12 indicate that MG33PB is effective against Gardonerella baginaris, and an inhibitory region is observed on the CBA plate (data not shown). Activity with either placebo MG33PB placebo (0%, 1%, and 2% BA) was observed, as indicated by the absence of inhibitory zones observed on the plate compared to the active formulation.</p><p num="0347"><u style="single">Summary</u> MG33PB is effective against Gardonerella baginalis compared to MTZ-free placebo gel.</p><p num="0348"> The disclosures of patents, patent documents, articles, abstracts, and other publications cited herein are incorporated herein by reference in their entirety, as if they were individually incorporated. In case of conflict, this specification, including the definitions, shall prevail. Various modifications and modifications to the present invention will be apparent to those skilled in the art without departing from the scope and gist of the present invention. Illustrated embodiments and examples are provided merely as examples and do not limit the scope of the invention. The scope of the invention is limited only by the claims set forth below.</p><p num="0349"> All publications, patents, patent applications, and other documents cited in this application may be incorporated by reference to their respective publications, patents, patent applications, or other documents for any purpose. To the same extent as indicated individually, the whole is incorporated herein by reference for any purpose.</p><p num="0350"> Various specific embodiments have been illustrated and described, but of course various modifications can be made without departing from the spirit and scope of the invention (s).</p>