Composition for peritoneal drug delivery and method of use thereof

Abstract

Compositions for the vaginal administration of an activator or salt to a subject are disclosed herein, comprising the activator or a salt thereof, a bioadhesive, and an emulsion. Methods for treating a disease condition by vaginal administration of a pharmaceutical composition to a subject, and kits comprising the pharmaceutical composition are provided herein. Further methods are provided for localizing an activator into the peritoneal cavity of a subject while minimizing systemic circulation of the activator. Provided herein are compositions, methods, and kits for delivering a drug into the peritoneal cavity. Provided herein are compositions comprising a cream, a polybioadhesive gel, or a liquid emulsion provided herein.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 442,054, filed on 30 January 2023, the contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] background Endometriosis is a condition in women characterized by the presence of endometrial tissue (endometrial glands and stroma) outside the uterine cavity, and is commonly associated with chronic pelvic pain and infertility. Examples of these lesions can be peritoneal lesions or deeply invasive diseases. Nearly 50% of infertile women suffer from endometriosis. Unfortunately, the effectiveness of treatments for endometriosis and other endometrial disorders includes hormonal therapy and analgesics to treat symptomatic endometriosis, which are limited, and endometriosis has a high recurrence rate in many women. Therefore, there is a great unmet need for treatments that can alleviate and prevent endometrial disease and related disorders in the long term, while reducing the systemic side effects associated with oral administration of current medications. [Overview of the project] [Means for solving the problem]

[0003] Brief overview Provided herein are compositions, methods, and kits for delivering drugs into the peritoneal cavity.

[0004] Provided herein are compositions comprising a cream, polybioadhesive gel, or liquid emulsion.

[0005] Provided herein is a cream comprising: (a) micronized danazol or a salt thereof present in an amount of 5.0% to about 7.0% by weight per volume (w / v) of the cream; (b) polyglycolized oleic acid glyceride present in an amount of 10% to 20% (w / v) of the cream; (c) cellulose polymer present in an amount of 1.0% to 3.0% (w / v) of the cream; and (d) one or more water-insoluble bioadhesives, wherein at least one water-insoluble bioadhesive is polycarbophil present in an amount of 1.0% to about 2.0% (w / v) of the cream, and the cream comprises at least 60% (w / v) water and one or more components selected from carbomer, methylparaben, sorbic acid, poloxamer, and sodium carboxymethylcellulose, and one or more water-insoluble bioadhesives.

[0006] Provided herein are compositions comprising a polybioadhesive gel containing: (a) an activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel, which, upon administration to a subject, causes the activator to localize in peritoneal tissue and induce pelvic transit metabolism in the subject.

[0007] Provided herein are compositions comprising the components shown in Table 3 and the respective amounts (w / v) of each component.

[0008] Provided herein is a method for treating a disease or condition of the endometrium of a subject, comprising administering a polybioadhesive gel transvaginally to the subject, comprising (a) an activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel, wherein the transvaginal administration of the polybioadhesive gel is a method for localizing the activator to the peritoneal tissue, thereby treating the disease or condition of the endometrium.

[0009] Provided herein is a method for inducing pelvic transit metabolism of an activator in a subject, comprising: (a) the activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel, wherein the vaginal administration increases the level of the activator in the peritoneal tissue relative to the level of the activator in the subject's serum, and maintains the ratio of the level of the activator in the subject's ascites fluid to the level of the activator in the subject's ascites fluid, wherein the vaginal administration is substantially the same as the oral administration of an oral pharmaceutical composition containing a substantially equivalent dose of the activator or a salt thereof, thereby localizing the activator to the peritoneal tissue and inducing pelvic transit metabolism in the subject.

[0010] Provided herein are methods for treating a disease or condition in a subject, comprising administering a cream or composition provided herein via vaginal delivery to the subject, thereby treating the disease or condition.

[0011] Provided herein is a method for maintaining ovarian function in a subject and reducing pain associated with endometriosis, comprising administering a cream or composition provided herein to the subject via vaginal delivery, thereby maintaining ovarian function in the subject and reducing pain associated with endometriosis.

[0012] Provided herein is a method for treating a disease or condition of the endometrium of a subject, comprising administering to the subject via vaginal administration a polybioadhesive gel comprising (a) an activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel, wherein the vaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or a salt thereof in the subject by at least 30 times compared to oral administration of the activator or a salt thereof to the subject; and the vaginal administration of the polybioadhesive gel localizes the activator to the peritoneal tissue of the subject, thereby treating the disease or condition of the endometrium of the subject.

[0013] Provided herein is a method for maintaining ovarian function in a subject and reducing pain associated with endometriosis, comprising administering to the subject via vaginal administration a polybioadhesive gel comprising (a) an activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel (thereby maintaining ovarian function in the subject and reducing pain associated with endometriosis), wherein the vaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or a salt thereof in the subject by at least 30 times compared to oral administration of the activator or a salt thereof to the subject; and the vaginal administration of the polybioadhesive gel localizes the activator to the peritoneal tissue of the subject.

[0014] Novel features are described in detail in the attached claims. A better understanding of the features and advantages can be obtained by referring to the following detailed description and attached drawings illustrating exemplary embodiments utilizing the exemplary principle. [Brief explanation of the drawing]

[0015] [Figure 1] Figure 1 shows a diagram of the peritoneal cavity and the organs contained within it.

[0016] [Figure 2] Figure 2 shows the VML-0501-001 study design and timeline.

[0017] [Figure 3] Figure 3 shows a graph of the ratio of danazol concentrations between serum and ascites in the (ITT) population. X-axis: condition; Y-axis: plasma ratio.

[0018] [Figure 4] Figure 4 shows a graph of the ratio of danazol concentrations between serum and ascites fluid in the PP population. X-axis: condition; Y-axis: plasma ratio.

[0019] [Figure 5]Figure 5 shows a graph of the concentration of danazol in the serum and tissues of the VML-0501 administration group. X-axis: conditions; Y-axis: concentration.

[0020] [Figure 6] Figure 6 shows a graph of the concentration of danazol in the serum and tissues of the oral danazol group. X-axis: conditions; Y-axis: concentration.

[0021] [Figure 7] Figure 7 shows a graph of the danazol concentration in the peritoneal tissue of the vaginal administration group. X-axis: conditions; Y-axis: concentration. N = 6. DETAILED DESCRIPTION OF THE INVENTION

[0022] Compositions and methods for transvaginal delivery of a drug are disclosed herein for reaching the pelvic region in a manner that can achieve high ascites and tissue concentrations at detectably low systemic circulation levels. In some cases, concentrations of at least 2 ng / mL, at least 5 ng / mL, at least >10 ng / mL or even higher can be achieved. Compared to other delivery methods (e.g., oral administration), the drug delivery methods disclosed herein can improve therapeutic efficacy. In some cases, such delivery can maintain a substantially zero-order release profile of a therapeutically effective amount of the active agent or its salt into ascites over a predetermined time interval when applied transvaginally to a subject.

[0023] Methods are provided herein for achieving a desired pharmacokinetic profile by transvaginal delivery of a pharmaceutical composition to a subject. In some cases, the pharmaceutical composition can include an emulsion that can be mixed with an active agent and a bioadhesive substance. In some embodiments, the bioadhesive substance is water-insoluble. In some embodiments, the water-insoluble bioadhesive substance is dispersed in at least a portion of the aqueous phase.

[0024] The vaginal administration of the pharmaceutical compositions described herein may be used to deliver the activator or a salt thereof locally into the peritoneal cavity. Using such delivery instead of systemic delivery may reduce the amount of the activator or salt present in circulation after vaginal administration of the pharmaceutical composition compared to systemic administration (e.g., oral administration) of the pharmaceutical composition, which may contain substantially equivalent doses of the activator or salt thereof. In some embodiments, the methods described herein may reduce the potential for systemic adverse events and undesirable side effects that may occur with systemic administration of the activator or salt thereof. Detection of the amount of the activator or salt thereof in a subject sample is also intended using an assay employing the detection of albumin conjugates by mass spectrometry.

[0025] Vaginal administration of a pharmaceutical composition may be used to treat a disease or condition in a subject. In some cases, the subject may be a subject who requires such treatment, for example, a subject who is suspected of having or has been previously diagnosed with a disease or condition treatable by vaginal administration of the pharmaceutical composition described herein. The pharmaceutical composition described herein may comprise at least one activator that can be selected based on the disease or condition to be treated.

[0026] A kit that may contain the pharmaceutical compositions described herein is provided herein. Such a kit may include instructions for application of the pharmaceutical composition and means for applying the pharmaceutical composition. definition

[0027] All definitions defined and used herein should be understood to take precedence over dictionary definitions, definitions in documents incorporated by reference, and / or the ordinary meanings of the defined terms.

[0028] As used herein and in the claims, the indefinite articles "a" and "an" should be understood to mean "at least one" unless explicitly stated otherwise.

[0029] As used herein and in the claims, the phrase “and / or” should be understood to mean “either or both” of the elements thus combined, that is, elements that exist associatively in some cases and disjunctively in others. Multiple elements enumerated by “and / or” should similarly be interpreted as “one or more” of the elements thus combined. Other elements may be present, as necessary, in addition to those specifically identified by the “and / or” clause, whether related to the specifically identified elements or not. Therefore, as a non-restrictive example, a reference to “A and / or B,” when used in combination with open-ended language such as “comprising,” may refer, in one embodiment, only A (including elements other than B as necessary); in another embodiment, only B (including elements other than A as necessary); in yet another embodiment, both A and B (including other elements as necessary); and so on.

[0030] Where used herein and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” should be interpreted as inclusive, that is, including at least one element but more than one of several elements or lists of elements, and, where applicable, additional unlisted items. Only terms explicitly indicated in contrast, such as “only one of” or “exactly one of” or, where used in the claims, “consisting of,” refer to the inclusion of exactly one element of several elements or lists of elements. In general, the term “or” as used herein shall be interpreted only as indicating an exclusive choice (i.e., “one or the other but not both”) when preceded by an exclusive term such as “either,” “one of,” “only one of,” or “exactly one of.” When used in claims, “consisting essentially of” shall have the usual meaning as used in the field of patent law.

[0031] Where used herein, “optional” or “optionally” means that the following circumstances may or may not occur, and as a result, the description includes both cases in which the circumstances occur and cases in which they do not.

[0032] As used herein, the terms “about” or “approximately” mean a range of up to ±20% of a given value. Alternatively, particularly with respect to biological systems or processes, the term may mean within one order of magnitude, preferably two times, of a given value. Where a particular value is stated in this application and claims, unless otherwise specified, the term “about” is implicit and in this context means within an acceptable margin of error for that particular value.

[0033] The terms "effective amount" or "therapeutic effective amount" refer to an amount sufficient to achieve, or at least partially achieve, the desired effect.

[0034] The term "zero-order release rate profile" or "zero-order release profile" may refer to a profile in which a given concentration of an activator can be released into the subject's vascular system at a constant rate over a given time interval. In some cases, the activator may have a substantially zero-order release profile into the subject's serum, lymphatic, or peritoneal vascular system upon administration of the pharmaceutical composition.

[0035] As used herein, the term "bioavailability" may refer to the extent to which a drug, such as an activator, salt, metabolite, or other substance, becomes available to target tissue after administration.

[0036] The parameters frequently used in pharmacokinetic (PK) studies are Tmax, Cmax, AUC(0-∞), AUC(0-t), and T 1 / 2 and may include CL / F. "Tmax" may refer to the time to reach the maximum plasma concentration ("Cmax") after administration of the therapeutic agent; "AUC(0-∞)" may refer to the area under the plasma concentration-time curve from time 0 to infinity; "AUC(0-t)" may refer to the area under the plasma concentration-time curve from time 0 to time t; "T 1 / 2 " may refer to the half-life of the therapeutic agent in plasma; "T 1 / 2,elim" may refer to the half-life of the drug's removal from circulation; and "CL / F" may refer to the apparent clearance rate of the therapeutic agent.

[0037] composition Provided herein are compositions for the topical delivery of activators or salts thereof. In some embodiments, the composition comprises an emulsion which may be a substantially homogeneous mixture of an organic phase and an aqueous phase, an activator or salt thereof, and a bioadhesion substance.

[0038] In some embodiments, the compositions provided herein may be formulated for transvaginal delivery. Formulation of the pharmaceutical composition may involve mixing an organic phase and an aqueous phase with a bioadhesive and an activator or a salt thereof. To improve the delivery of the activator or a salt thereof, a bioadhesive may be used to adhere the pharmaceutical composition to the epithelial surface upon transvaginal administration.

[0039] The inventors have discovered a surprising and unexpected result: when pharmaceutical compositions described herein, comprising a liquid emulsion, are formulated with a bioadhesive substance by emulsifying an insoluble substance, they can prevent or minimize vaginal aggregates or discharges, thereby minimizing or eliminating unsightly aggregates or discharges. Such emulsions may comprise a substantially homogeneous mixture of an organic phase containing an activator or a salt thereof and a bioadhesive substance, and an aqueous phase. The aqueous and organic phases may be provided as a homogeneous mixture, or each component may be provided separately for mixing before administration.

[0040] Emulsions containing an organic phase and an aqueous phase may contain a variety of components or components. In some cases, the organic phase may contain at least one oleogel. The oleogel may contain at least one oily agent and at least one polymer.

[0041] The oily agents may be monoglycerides, diglycerides, triglycerides, or any combination thereof. In some cases, the oily agents may be separated and purified. In some cases, the oily agents may be synthetic diglycerides, synthetic triglycerides, propylene glycol isostearate, polyoxyethylene-derived oleic acid glyceride mixtures, plant or natural oils, or any combination thereof.

[0042] In some cases, the amount of oily agent is approximately 0.1-1%, 0.1-2%, 0.1-3%, 0.1-4%, 0.1-5%, 0.1-6%, 0.1-7%, 0.1-8%, 0.1-9%, 0.1-10%, 0.1-11%, 0.1-12%, 0.1-13%, 0.1-14%, 0.1- They may be present in proportions of approximately 15%, 0.1-16%, 0.1-17%, 0.1-18%, 0.1-19%, 0.1-20%, 0.1-21%, 0.1-22%, 0.1-23%, 0.1-24%, 0.1-25%, 0.1-26%, 0.1-27%, 0.1-28%, 0.1-29%, or 0.1-30%.

[0043] In some cases, the oily agent is added in amounts of approximately 0.01-0.1%, 0.01-0.2%, 0.01-0.3%, 0.01-0.4%, 0.01-0.5%, 0.01-0.6%, 0.01-0.7%, 0.01-0.8%, 0.01-0.9%, 0.01-1%, 0.01-2%, 0.01-3%, 0.01-4%, 0.01-5%, 0.01-6%, 0.01-7%, 0.01-8%, 0.01-9%, and 0.01- They may exist in proportions of 10%, 0.01-approximately 11%, 0.01-approximately 12%, 0.01-approximately 13%, 0.01-approximately 14%, 0.01-approximately 15%, 0.01-approximately 16%, 0.01-approximately 17%, 0.01-approximately 18%, 0.01-approximately 19%, 0.01-approximately 20%, 0.01-approximately 21%, 0.01-approximately 22%, 0.01-approximately 23%, 0.01-approximately 24%, 0.01-approximately 25%, 0.01-approximately 26%, 0.01-approximately 27%, 0.01-approximately 28%, 0.01-approximately 29%, or 0.01-approximately 30%.

[0044] In some exemplary embodiments, the oily agent may be propylene glycol isostearate. Exemplary pharmaceutical compositions may contain propylene glycol isostearate in an amount of about 5 to 90% by weight relative to the total weight of the oleogel.

[0045] Monoglycerides, diglycerides, or triglycerides are molecules of formula I: [ka] (In the formula, R1, R2, and R3 are independently H; or C1-C containing a degree of unsaturation of 0, 1, 2, 3, 4, or 5) 20 (It could be alkyl.)

[0046] In some embodiments, the synthetic monoglycerides, diglycerides, or triglycerides may be "LABRAFAC® lipophile WL1349" sold by Gatefosse, propylene glycol isostearate, for example, a product sold by Gatefosse under the name "hydrophilol isostearique," and polyglycolized glycerides "LABRRAFIL® M 1944 CS" sold by Gatefosse.

[0047] LABRRAFIL® M 1944 CS is a mixture of polyoxyethylene-derived oleic acid glycerides obtained by the alcoholic decomposition of natural vegetable oils. It may be an oily liquid whose properties are shown in Table 1 below. [Table 1]

[0048] In some cases, monoglycerides, diglycerides, or triglycerides may be of natural or plant origin. Examples of natural or plant-derived oils include sweet almond oil, argan oil, or palm oil.

[0049] The polymer in the organic phase may be a cellulose polymer. In some cases, the cellulose polymer may be ethylcellulose, sodium-free carboxymethylcellulose, or a mixture thereof. In some cases, the polymer may be a water-insoluble polymer. In some exemplary embodiments, the water-insoluble polymer may be a water-insoluble cellulose polymer.

[0050] The cellulose polymer may be a lipid-soluble cellulose polymer. In some cases, the cellulose polymer may be alkylcellulose. In some cases, the alkylcellulose may be methylcellulose, ethylcellulose, hydroxypropylcellulose, or a combination thereof. In some embodiments, the cellulose polymer may be alkylcarboxylic acid-containing cellulose or a salt thereof. In some cases, the alkylcarboxylic acid-containing cellulose may be sodium-free carboxymethylcellulose.

[0051] In some cases, the water-insoluble polymer is present in amounts of approximately 0.1–1%, 0.1–2%, 0.1–3%, 0.1–4%, 0.1–5%, 0.1–6%, 0.1–7%, 0.1–8%, 0.1–9%, 0.1–10%, 0.1–11%, 0.1–12%, 0.1–13%, 0.1–14%, and 0. They may be present in proportions of 1-15%, 0.1-16%, 0.1-17%, 0.1-18%, 0.1-19%, 0.1-20%, 0.1-21%, 0.1-22%, 0.1-23%, 0.1-24%, 0.1-25%, 0.1-26%, 0.1-27%, 0.1-28%, 0.1-29%, or 0.1-30%.

[0052] In some cases, the water-insoluble polymer is present in amounts of approximately 0.01-0.1%, 0.01-0.2%, 0.01-0.3%, 0.01-0.4%, 0.01-0.5%, 0.01-0.6%, 0.01-0.7%, 0.01-0.8%, 0.01-0.9%, 0.01-1%, 0.01-2%, 0.01-3%, 0.01-4%, 0.01-5%, 0.01-6%, 0.01-7%, 0.01-8%, 0.01-9%, and 0.0% of the weight of the composition. They may exist in proportions of 1-10%, 0.01-11%, 0.01-12%, 0.01-13%, 0.01-14%, 0.01-15%, 0.01-16%, 0.01-17%, 0.01-18%, 0.01-19%, 0.01-20%, 0.01-21%, 0.01-22%, 0.01-23%, 0.01-24%, 0.01-25%, 0.01-26%, 0.01-27%, 0.01-28%, 0.01-29%, or 0.01-30%.

[0053] In some exemplary embodiments, the cellulose polymer may be present in a proportion of 1 to about 10% by weight. In some exemplary embodiments, the cellulose polymer may be ethylcellulose present in a proportion of 1 to about 10% by weight based on the total weight of the composition. In some cases, the oily agent may consist of LABRRAFIL® M1944CS. In some cases, the oily agent may be present in a proportion of about 5 to 90% by weight relative to the total weight of the oleogel. In some cases, the ratio of oleogel to aqueous gel weight may be about 10:90 to about 90:10. In some cases, the cellulose polymer may be EMULFREE® P. In some embodiments, the cellulose polymer may be EMULFREE® P, and the oily agent may consist of LABRRAFIL® M1944CS.

[0054] The aqueous phase described herein may comprise one or more aqueous gels. The aqueous phase may comprise at least one aqueous gel. In some cases, the aqueous gel may comprise at least one gelling agent. The gelling agent may be carbomer, poloxamer, sodium carboxymethylcellulose, or any mixture thereof.

[0055] In some embodiments, the gelling agent is present in amounts of approximately 0.1 to 1%, 0.1 to 2%, 0.1 to 3%, 0.1 to 4%, 0.1 to 5%, 0.1 to 6%, 0.1 to 7%, 0.1 to 8%, 0.1 to 9%, 0.1 to 10%, 0.1 to 11%, 0.1 to 12%, 0.1 to 13%, 0.1 to 14%, and 0.1% of the weight of the aqueous phase. They may exist in proportions of ~15%, 0.1~16%, 0.1~17%, 0.1~18%, 0.1~19%, 0.1~20%, 0.1~21%, 0.1~22%, 0.1~23%, 0.1~24%, 0.1~25%, 0.1~26%, 0.1~27%, 0.1~28%, 0.1~29%, or 0.1~30%.

[0056] In some cases, the gelling agent is added in amounts of approximately 0.01-0.1%, 0.01-0.2%, 0.01-0.3%, 0.01-0.4%, 0.01-0.5%, 0.01-0.6%, 0.01-0.7%, 0.01-0.8%, 0.01-0.9%, 0.01-1%, 0.01-2%, 0.01-3%, 0.01-4%, 0.01-5%, 0.01-6%, 0.01-7%, 0.01-8%, 0.01-9%, 0.01- They may exist in proportions of approximately 10%, 0.01-11%, 0.01-12%, 0.01-13%, 0.01-14%, 0.01-15%, 0.01-16%, 0.01-17%, 0.01-18%, 0.01-19%, 0.01-20%, 0.01-21%, 0.01-22%, 0.01-23%, 0.01-24%, 0.01-25%, 0.01-26%, 0.01-27%, 0.01-28%, 0.01-29%, or 0.01-30%.

[0057] In some embodiments, the gelling agent for the aqueous phase may be a carbomer, Carbopol974, or Carbopol980, present in a proportion of about 0.1 to about 5% by weight relative to the total weight of the aqueous phase.

[0058] In some cases, the ratio of the weight of the organic phase to the weight of the aqueous phase may be about 1:9 to about 9:1, about 1:8 to about 8:1, about 1:7 to about 7:1, about 1:6 to about 6:1, about 1:5 to about 5:1, about 1:4 to about 4:1, about 1:3 to about 3:1, about 1:2 to about 2:1, or about 1:1.5 to about 1.5:1. In some cases, the ratio of the weight of the organic phase to the weight of the aqueous phase may be about 1:1. In some embodiments, the emulsion in the compositions herein may comprise a substantially homogeneous mixture of the organic phase and the aqueous phase. In some cases, the ratio of the weight of the organic phase to the weight of the aqueous phase in such a substantially homogeneous mixture may be about 1:1.

[0059] In some cases, a homogeneous mixture of an organic phase and an aqueous phase, when stored in a sealed container, may maintain a stable, uniform appearance for a period of time. In some cases, the mixture may be stable for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, or 52 weeks. In some cases, the mixture may be stable for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 months. In some cases, the mixture may be stable for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years. In some cases, the sealed container may be stored at a temperature of about 25°C and about 1 atmosphere. In some cases, the sealed container may be stored at a relative humidity of about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, or 100%.

[0060] In some cases, the ratio of the volume of the organic phase to the volume of the aqueous phase can be approximately 1:9 to 9:1, 1:8 to 8:1, 1:7 to 7:1, 1:6 to 6:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, 1:2 to 2:1, or 1:1.5 to 1.5:1. In some cases, the ratio of the volume of the organic phase to the volume of the aqueous phase can be approximately 1:1.

[0061] When a water-immiscible oil phase can be mixed with an aqueous phase, the emulsion may include dispersions or droplets, as well as other lipid structures that can be formed as a result of hydrophobic forces that separate polar residues (e.g., long hydrocarbon chains) from water and orient polar head groups toward water. These other lipid structures may include monolayers, multilayers, multilayer lipid vesicles, micelles, and lamellar phases.

[0062] In some cases, the penetration enhancer may be present in an amount sufficient to achieve an increased penetration rate. In some cases, the penetration enhancer does not enhance the penetration rate of the activator or its salt below a threshold concentration. The threshold concentration may be specific to a given penetration enhancer. In some cases, the penetration enhancer is present in an amount of at least about 0.1% by weight, 0.11% by weight, 0.12% by weight, 0.13% by weight, 0.14% by weight, 0.15% by weight, 0.16% by weight, 0.17% by weight, 0.18% by weight, 0.19% by weight, 0.2% by weight, 0.21% by weight, 0.22% by weight, 0.23% by weight, 0.24% by weight, 0.25% by weight, 0.26% by weight, 0.27% by weight, 0.28% by weight, 0.29% by weight, 0.3% by weight, 0.31% by weight, 0.32% by weight, 0.33% by weight, 0.34% by weight, 0.35% by weight, 0.36% by weight, 0.37% by weight, 0.38% by weight, 0.39% by weight, 0.4% by weight %, 0.41 wt%, 0.42 wt%, 0.43 wt%, 0.44 wt%, 0.45 wt%, 0.46 wt%, 0.47 wt%, 0.48 wt%, 0.49 wt%, 0.5 wt%, 0.51 wt%, 0.52 Weight%, 0.53% by weight, 0.54% by weight, 0.55% by weight, 0.56% by weight, 0.57% by weight, 0.58% by weight, 0.59% by weight, 0.6% by weight, 0.61% by weight, 0.62% by weight, 0.63% by weight, 0 .64% by weight, 0.65% by weight, 0.66% by weight, 0.67% by weight, 0.68% by weight, 0.69% by weight, 0.7% by weight, 0.71% by weight, 0.72% by weight, 0.73% by weight, 0.74% by weight, 0.75% by weight , 0.76 wt%, 0.77 wt%, 0.78 wt%, 0.79 wt%, 0.8 wt%, 0.81 wt%, 0.82 wt%, 0.83 wt%, 0.84 wt%, 0.85 wt%, 0.86 wt%, 0.87 wt%, 0.88 wt%, 0.89 wt%, 0.9 wt%, 0.91 wt%, 0.92 wt%, 0.93 wt%, 0.94 wt%, 0.95 wt%, 0.96 wt%, 0.97 wt%, 0.98 wt%, 0.99%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20 Weight%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 3 It promotes the absorption rate of the activator or its salt when present at weight concentrations of 9% by weight, 40% by weight, 41% by weight, 42% by weight, 43% by weight, 44% by weight, 45% by weight, 46% by weight, 47% by weight, 48% by weight, 49% by weight, 50% by weight, 51% by weight, 52% by weight, 53% by weight, 54% by weight, 55% by weight, 56% by weight, 57% by weight, 58% by weight, 59% by weight, 60% by weight, 61% by weight, 62% by weight, 63% by weight, 64% by weight, 65% by weight, 66% by weight, 67% by weight, 68% by weight, 69% by weight, or 70% by weight.

[0063] In some cases, the compositions described herein do not contain penetration enhancers. In some cases, the compositions described herein contain about 0.1% by weight, 0.11% by weight, 0.12% by weight, 0.13% by weight, 0.14% by weight, 0.15% by weight, 0.16% by weight, 0.17% by weight, 0.18% by weight, 0.19% by weight, 0.2% by weight, 0.21% by weight, 0.22% by weight, 0.23% by weight, 0.24% by weight, 0.25% by weight, 0.26% by weight, 0.27% by weight, and 0.28% by weight, relative to the total weight of the composition. %, 0.29% by weight, 0.3% by weight, 0.31% by weight, 0.32% by weight, 0.33% by weight, 0.34% by weight, 0.35% by weight, 0.36% by weight, 0.37% by weight, 0.38% by weight, 0.39% by weight, 0.4% by weight Amount %, 0.41 wt%, 0.42 wt%, 0.43 wt%, 0.44 wt%, 0.45 wt%, 0.46 wt%, 0.47 wt%, 0.48 wt%, 0.49 wt%, 0.5 wt%, 0.51 wt%, 0.5 2% by weight, 0.53% by weight, 0.54% by weight, 0.55% by weight, 0.56% by weight, 0.57% by weight, 0.58% by weight, 0.59% by weight, 0.6% by weight, 0.61% by weight, 0.62% by weight, 0.63% by weight, 0.64% by weight, 0.65% by weight, 0.66% by weight, 0.67% by weight, 0.68% by weight, 0.69% by weight, 0.7% by weight, 0.71% by weight, 0.72% by weight, 0.73% by weight, 0.74% by weight, 0.75% by weight %, 0.76 wt%, 0.77 wt%, 0.78 wt%, 0.79 wt%, 0.8 wt%, 0.81 wt%, 0.82 wt%, 0.83 wt%, 0.84 wt%, 0.85 wt%, 0.86 wt%, 0.87 wt%, 0.88 wt%, 0.89 wt%, 0.9 wt%, 0.91 wt%, 0.92 wt%, 0.93 wt%, 0.94 wt%, 0.95 wt%, 0.96 wt%, 0.97 wt%, 0.98 wt%, 0.99%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% Amount%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37% It contains a penetration enhancer in an amount not exceeding %, 38% by weight, 39% by weight, 40% by weight, 41% by weight, 42% by weight, 43% by weight, 44% by weight, 45% by weight, 46% by weight, 47% by weight, 48% by weight, 49% by weight, 50% by weight, 51% by weight, 52% by weight, 53% by weight, 54% by weight, 55% by weight, 56% by weight, 57% by weight, 58% by weight, 59% by weight, 60% by weight, 61% by weight, 62% by weight, 63% by weight, 64% by weight, 65% by weight, 66% by weight, 67% by weight, 68% by weight, 69% by weight, or 70% by weight.

[0064] The oleogel and aqueous phase may each further contain, in conventional amounts known not to cause skin irritation, standard components for various types of gels, such as texturers, antioxidants, preservatives, dyes, or fragrances.

[0065] The composition may be in the form of a pharmaceutical composition. In some embodiments, the pharmaceutical composition may be administered similarly to cosmetics in the form of a cream or gel that can be applied to the skin. In some embodiments, the composition may be in the form of a unit dosage form, when applied in a specified amount to at least a portion of the skin. Since the pharmaceutical composition may contain a stable mixture of an oleogel and an aqueous gel, it may be free of clumps and moisture, require a smaller application area compared to conventional aqueous alcohol gels, and be fast-drying.

[0066] In some embodiments, the gel composition may contain bioadhesives. Examples of bioadhesives include carbomer, glyceryl monooleate, hypromellose, polycarbophil, poly(methyl vinyl ether-co-maleic anhydride), and their salts. In some cases, the pharmaceutical composition may contain one or more bioadhesives. In some cases, the pharmaceutical composition may contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bioadhesives.

[0067] In some exemplary embodiments, the bioadhesive material may be polycarbophil or a salt thereof. Polycarbophil is designed to mimic loaded mucin, a glycoprotein component of mucus involved in adhesion to the underlying epithelial surface. Polycarbophil is a lightly crosslinked polymer. Polycarbophil is also a weak polyacid containing multiple carboxyl radicals (COO-), which are its source of negative charge. These acid radicals enable hydrogen bonding with the cell surface. While hydrogen bonding can be weak, in the case of polycarbophil, they can be numerous. Bioadhesive materials such as polycarbophil can maintain their adhesion to vaginal epithelial cells until turnover (which can be up to 7 days in menopausal women). However, non-water-soluble polycarbophil that remains attached to vaginal epithelial cells can lead to vaginal aggregates and vaginal discharge. Therefore, it is necessary to provide formulations that can provide adhesion of a pharmaceutical composition to the epithelium of a subject while minimizing vaginal aggregates or vaginal discharge.

[0068] In some cases, bioadhesive substances make up at least about 0.01%, at least about 0.05%, at least about 0.1%, at least about 0.15%, at least about 0.2%, at least about 0.25%, at least about 0.3%, at least about 0.35%, at least about 0.4%, at least about 0.45%, at least about 0.5%, at least about 0.55%, at least about 0.6%, at least about 0.65%, at least about 0.7%, at least about 0.75%, at least about 0.8%, at least about 0.85%, at least about 0.9%, at least about 0.95%, at least about 1%, at least about 1.1%, and a small amount. It may be present at a weight concentration of at least approximately 1.2%, at least approximately 1.3%, at least approximately 1.4%, at least approximately 1.5%, at least approximately 1.6%, at least approximately 1.7%, at least approximately 1.8%, at least approximately 1.9%, at least approximately 2%, at least approximately 2.5%, at least approximately 3%, at least approximately 3.5%, at least approximately 4%, at least approximately 4.5%, at least approximately 5%, at least approximately 5.5%, at least approximately 6%, at least approximately 6.5%, at least approximately 7%, at least approximately 7.5%, at least approximately 8%, at least approximately 8.5%, at least approximately 9%, at least approximately 9.5%, or at least approximately 10%. In some embodiments, the aqueous gel may contain polycarbophil at weight concentrations of about 0.1% to about 10%, about 0.1% to about 9%, about 0.1% to about 8%, about 0.1% to about 7%, about 0.1% to about 6%, about 0.1% to about 5%, about 0.1% to about 4%, about 0.1% to about 3%, about 0.1% to about 2%, about 0.1% to about 1%, about 0.1% to about 0.9%, about 0.1% to about 0.8%, about 0.1% to about 0.7%, about 0.1% to about 0.6%, about 0.1% to about 0.5%, about 0.1% to about 0.4%, about 0.1% to about 0.3%, or about 0.1% to about 0.2%.

[0069] The composition may contain alcohols. For example, the alcohol may be a C1-C8 alcohol. Examples of C1-C8 alcohols include methanol, ethanol, n-propanol, isopropanol, t-butanol, pentanol, hexanol, cyclohexanol, heptanol, and octanol.

[0070] Activating agent The pharmaceutical compositions described herein may comprise at least one activator or a salt thereof. While exemplary activators are described herein, further activators in the compositions may be substituted to treat other indications treatable by administration of the pharmaceutical composition to a subject.

[0071] In some embodiments, the active ingredient may be a hormone, an anti-inflammatory agent, an analgesic, phenethylamine, an antineoplastic agent, a steroid, a 5-α-reductase inhibitor, a gonadotropin-releasing hormone (GnRH) agonist, a GnRH antagonist, a glycine receptor antagonist, tetrahydrocannabinol, analgesics; an antibiotic, an antiviral compound, an antifungal compound, a salt of any of these, or any combination thereof.

[0072] In some embodiments, the hormone may be testosterone; dihydrotestosterone (DHT); estradiol; ethinylestradiol; progesterone; levonorgestrel; desogestrel; peptide hormones, such as oxytocin; synthetic progesterone; salts of any of these or any combination thereof.

[0073] In some embodiments, the active ingredient may be an analgesic. Examples of analgesics include acetaminophen, bromfenac, diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, meclofenamete, mefenamic acid, meloxicam, nabumetone, naproxen, nepafenac, oxaprozine, phenylbutazone, piroxicam, sulindac, tolmetine, celecoxib, buprenorphine, butorphanol, codeine, hydrocodone, hydromorphone, levorphanol, meperidine, methadone, morphine, nalbuffine, oxycodone, oxymorphone, pentazocine, propoxifene, tapentadol, tramadol, aspirin, any salt thereof, or any combination thereof.

[0074] In some embodiments, phenethylamine may be dopamine, epinephrine, norepinephrine, phenylephrine, methylphenidate, amphetamine, any salt thereof, or any combination thereof.

[0075] In some embodiments, the antineoplastic agent may be cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, procarbazine, prednisolone, bleomycin, vinblastine, dacarbazine, cisplatin, epirubicin, dichloroacetate, any salt thereof, or any combination thereof.

[0076] In some embodiments, the steroid may be danazol or a salt thereof. Danazol is a synthetic androgen; a derivative of the synthetic steroid ethisterone. Chemically, danazol, USP is 17α-pregna-2,4-dien-20-ino[2,3-d]-isoxazole-17-ol.

[0077] In some embodiments, the 5-α-reductase inhibitor may be dutasteride, tamsulosin, finasteride, a salt of any of these, or any combination thereof.

[0078] In some embodiments, the GnRH agonist, GnRH antagonist may be leuprolide, buserelin, histrelin, goserelin, deslorerin, nafarelin, triptorelin, cetrorelix, abarelix; ganirelix, ozarelix, degarelix or teverelix, any salt thereof, or any combination thereof.

[0079] In some embodiments, the GnRH antagonist may be Cetrorelics, Abarerlics; Ganirellics, Ozarellics, Degarellics, Tevererlics, any salt thereof, or any combination thereof.

[0080] In some embodiments, the glycine receptor antagonist may be tranexamic acid or a salt thereof.

[0081] In some embodiments, the antibiotic may be ceftobiprole; cephthaloline; clindamycin; dalbavancin; daptomycin; linezolid; mupirosin; oritabancin; tedizolid; teravancin; tigecycline; vancomycin; aminoglycosides; carbapenems; ceftazidime; cefepime; ceftobiprole; fluoroquinolones; piperacillin; ticalcillin; linezolid; streptogramin; tigecycline; daptomycin; salts of any of these; or any combination thereof.

[0082] In some embodiments, the antiviral compound may be ceftobiprole; cephthaloline; clindamycin; dalbavancin; daptomycin; linezolid; mupirosin; oritabancin; tedizolid; teravancin; tigecycline; vancomycin; aminoglycoside; carbapenem; ceftazidime; cefepime; ceftobiprole; fluoroquinolone; piperacillin; ticalcillin; linezolid; streptogramin; tigecycline; daptomycin; salts of any of these; or any combination thereof.

[0083] In some embodiments, the antifungal compound may be cyclopiroxolamine; haloprozine; tolnaphthate; undecylenate; topical nystatin; amorolfine; butenafine; naphthifine; terbinafine; or any combination thereof.

[0084] In some embodiments, the composition may contain an activator or a salt thereof in amounts not exceeding approximately 10 mg, not exceeding approximately 20 mg, not exceeding approximately 40 mg, not exceeding approximately 60 mg, not exceeding approximately 80 mg, not exceeding approximately 100 mg, not exceeding approximately 120 mg, not exceeding approximately 140 mg, not exceeding approximately 160 mg, not exceeding approximately 180 mg, not exceeding approximately 200 mg, not exceeding approximately 300 mg, not exceeding approximately 400 mg, or not exceeding approximately 500 mg. In some cases, the composition may contain at least approximately 10 mg, at least approximately 20 mg, at least approximately 40 mg, at least approximately 60 mg, at least approximately 80 mg, at least approximately 100 mg, at least approximately 120 mg, at least approximately 140 mg, at least approximately 160 mg, at least approximately 180 mg, at least approximately 200 mg, at least approximately 300 mg, at least approximately 400 mg, or at least approximately 500 mg of an activator or a salt thereof.

[0085] The activator may constitute a portion of the total weight of the composition. In some cases, the activator may constitute about 0.000001% to about 99% by weight, about 0.000001% to about 50% by weight, about 0.000001% to about 30% by weight, about 0.000001% to about 20% by weight, about 0.00005% to about 15% by weight, about 0.00005% to about 10% by weight, about 0.00001% to about 10% by weight, about 0.0001% to about 10% by weight, or about 0.0001% to about 5% by weight of the total weight of the composition.

[0086] In some embodiments, the composition may contain alcohol at a concentration that enables the solubilization of the activator. In some cases, the composition may have an alcohol concentration of at most about 10% by weight, at most about 8% by weight, at most about 6% by weight, at most about 5% by weight, at most about 4% by weight, at most about 3% by weight, at most about 2% by weight, or at most about 1% by weight. In some cases, the alcohol concentration is about 3.5% by weight.

[0087] In some embodiments, the composition may be substantially free of surfactants. In some cases, the composition may contain small amounts of surfactants. The surfactant may be a nonionic surfactant, a cationic surfactant, an amphoteric surfactant, or an amphoteric surfactant.

[0088] In some cases, the composition may have a surfactant concentration of at most about 10%, at most about 8%, at most about 6%, at most about 5%, at most about 4%, at most about 3%, at most about 2%, at most about 1%, or at most about 0.1% of the total weight of the composition.

[0089] If the composition contains an activator or a salt thereof, the activator or a salt thereof may be emulsified in an emulsion. When emulsified in an emulsion, the activator may be in a form having an average particle size minimized to about a micrometer scale. In some embodiments, the average particle size may be minimized to about a nanometer scale.In some cases, the average particle size is approximately 0.001 nm to 500 μm, approximately 0.001 nm to 400 μm, approximately 0.001 nm to 300 μm, approximately 0.001 nm to 200 μm, approximately 0.001 nm to 100 μm, approximately 0.001 nm to 90 μm, approximately 0.001 nm to 80 μm, approximately 0.001 nm to 70 μm, approximately 0.001 nm to 60 μm, approximately 0.001 nm to 50 μm, approximately 0.001 nm to 40 μm, approximately 0.001 nm to 30 μm, approximately 0.001 nm to 20 μm, approximately 0.001nm to about 10μm, about 0.001nm to about 5μm, about 0.001nm to about 1μm, about 0.001nm to about 900nm, about 0.001nm to about 800nm, about 0.001nm to about 700nm, about 0.001nm to about 600nm, about 0. 001nm ~ approx. 500nm, approx. 0.001nm ~ approx. 400nm, approx. 0.001nm ~ approx. 300nm, approx. 0.001nm ~ approx. 200nm, approx. 0.001nm ~ approx. 100nm, approx. 0.001nm ~ approx. 90nm, approx. 0.001nm ~ approx. .001nm to about 70nm, about 0.001nm to about 60nm, about 0.001nm to about 50nm, about 0.001nm to about 40nm, about 0.001nm to about 30nm, about 0.001nm to about 20nm, about 0.001nm to about 10nm, about 0.00 1nm to about 5nm, about 0.001nm to about 1nm, about 0.001nm to about 0.9nm, about 0.001nm to about 0.8nm, about 0.001nm to about 0.7nm, about 0.001nm to about 0.6nm, about 0.001nm to about 0.5nm, about 0.001 nm to approximately 0.4nm, approximately 0.001nm to approximately 0.3nm, approximately 0.001nm to approximately 0.2nm, approximately 0.001nm to approximately 0.1nm, approximately 0.001nm to approximately 0.09nm, approximately 0.001nm to approximately 0.08nm, approximately 0.001nm to approximately 0.07nm, approximately 0.001nm to approximately 0.06nm, approximately 0.001nm to approximately 0.05nm, approximately 0.001nm to approximately 0.04nm, approximately 0.001nm to approximately 0.03nm, approximately 0.001nm to approximately 0.02nm, or approximately 0.001nm to approximately 0.01nm.In some embodiments, the average particle size is approximately 0.01 nm, 0.05 nm, 0.1 nm, 0.15 nm, 0.2 nm, 0.25 nm, 0.3 nm, 0.35 nm, 0.4 nm, 0.45 nm, 0.5 nm, 0.55 nm, 0.6 nm, 0.65 nm, 0.7 nm, 0.75 nm, 0.8 nm, 0.85 nm, 0.9 nm, and 0.95 nm. m, about 1nm, about 2nm, about 3nm, about 4nm, about 5nm, about 6nm, about 7nm, about 8nm, about 9nm, about 10nm, about 15nm, about 20nm, about 25nm, about 30nm, about 35nm, about 40nm, about 45nm, about 50nm, about 55nm, about 60nm, about 65nm, about 70nm, about 75nm, about 80nm, about 85nm, about 90nm, about 95nm, about 100nm, about 150nm , about 200nm, about 250nm, about 300nm, about 350nm, about 400nm, about 450nm, about 500nm, about 550nm, about 600nm, about 650nm, about 700nm, about 750nm , about 800nm, about 850nm, about 900nm, about 950nm, about 1μm, about 2μm, about 3μm, about 4μm, about 5μm, about 6μm, about 7μm, about 8μm, about 9μm, about 10μm, about 15μ m may be approximately 20 μm, 25 μm, 30 μm, 35 μm, 40 μm, 45 μm, 50 μm, 55 μm, 60 μm, 65 μm, 70 μm, 75 μm, 80 μm, 85 μm, 90 μm, 95 μm, 100 μm, 150 μm, 200 μm, 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, or 500 μm.

[0090] Treatment method The methods and compositions described herein may be used to treat conditions or diseases. In some cases, treatment of a disease may include at least partially improving at least one symptom of the disease or condition. Examples of diseases or conditions that may be treated with the pharmaceutical compositions described herein include endometrial disorders, adenomyosis, cancer, inflammatory disorders, infections, and any combination thereof. While exemplary diseases or conditions are listed herein, those skilled in the art may treat further diseases or conditions by substituting additional activators with the pharmaceutical compositions described herein.

[0091] In some embodiments, the disease or condition may be an endometrial disorder. For example, an endometrial disorder may be endometriosis. Endometriosis can be a frequently painful disorder in which tissue that is normally located inside the uterus (e.g., endometrium) grows outside the uterus (endometrial implants). The methods and compositions may be used to treat endometriosis involving the ovaries, intestines, or pelvic endometrial tissue, or endometriosis in which endometrial tissue has spread outside the pelvic region. In endometriosis, the displaced deposits break down and bleed with each menstrual cycle. Since this displaced tissue has no way to leave the body, it can become trapped. The surrounding tissue becomes inflamed and can eventually lead to scar tissue and adhesions (abnormal tissue that binds organs together). In some cases, treatment of the condition may involve reducing the amount of endometrial deposits to a level that may be less than before treatment.

[0092] In some cases, endometrial disorders can be adenomyosis. Adenomyosis is a disorder in which the inner wall of the uterine lining penetrates the muscular wall of the uterus (myometrium). Adenomyosis can cause painful menstruation, decreased intra-abdominal pressure, and bloating before menstruation, resulting in a very difficult period. The condition can affect the entire uterus or be limited to one location. The cause of adenomyosis is unknown, but research suggests that various hormones, including estrogen, progesterone, prolactin, and follicle-stimulating hormone, may trigger the condition. Current treatments include administering anti-inflammatory drugs to reduce inflammation; and administering therapeutic interventions such as aromatase inhibitors, GnRH agonists, or GnRH antagonists to suppress the expression of hormones that may trigger the condition. In some cases, the development of adenomyosis and endometriosis can occur simultaneously.

[0093] Treatment of endometrial disorders using the vaginal compositions described herein can be determined using many metrics known in the art. Treatment endpoints for endometrial disorders may include fertility. In some cases, fertility can be determined using in vitro assays such as human chorionic gonadotropin (hCG) assays. hCG assays may be performed on subject-derived samples, such as blood samples. Such assays can determine blood hCG levels over time using hCG-specific antibodies that can be used to determine the incidence of pregnancy. Fertility can also be determined by determining the presence of mature oocytes in the subject or the success of zygote implantation at fertilization using in vivo imaging means such as ultrasound.

[0094] In some embodiments, the disease or condition may be cancer. For example, the cancer may be cancer of the reproductive tract, cervical cancer, ovarian cancer, mesothelial cancer, peritoneal cancer, or any combination thereof.

[0095] In exemplary embodiments, the vaginal administration of the pharmaceutical compositions described herein may be used to locally treat cancer in the peritoneal cavity. The vaginal administration of the pharmaceutical compositions described herein may be used to minimize adverse side effects that may occur with systemic administration. For example, the pharmaceutical composition containing the antineoplastic agent may be used to deliver the antineoplastic agent directly to the peritoneal cavity, with minimal delivery of the neoadjuvant to the circulatory system. As an exemplary example, such vaginal administration of the pharmaceutical composition may be used to mitigate known side effects of systemic antineoplastic agent administration, such as hair loss, reproductive side effects, skin or nail inflammation, swelling, cardiotoxicity, and hepatotoxicity. Reduction of side effects may be determined by methods such as monitoring the incidence of side effect symptoms. Examples include reduction of local irritation or inflammation, reduction of vomiting incidence, reduction of pain incidence, reduction of irregular heart rate or arrhythmia, and reduction of frequent or painful urination. Biomarkers may also be used to measure the reduction of side effects. In some cases, reduction of side effects may include a decrease in toxicity-related biomarkers. In some cases, a reduction in side effects may include an increase in toxicity-related biomarkers. Examples of biomarkers include cardiac troponin I, cardiac troponin T, serum alanine aminotransferase, glutathione-S-transferase α, aspartate aminotransferase, serum creatinine, blood urea nitrogen, kidney injury molecule-1, neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), cystatin C, clatherin, fatty acid-binding protein-liver type (L-FABP), and osteopontin.

[0096] In some cases, cancer treatment may include reducing tumor size or slowing or preventing tumor growth. Imaging techniques, such as ultrasound or magnetic resonance imaging (MRI), can be used to determine the tumor volume of a subject, and this can be compared over time to determine the therapeutic effect of administering a pharmaceutical composition over time.

[0097] In some embodiments, the condition may be an inflammatory disorder. For example, inflammatory diseases may be pelvic inflammatory disease, chronic pelvic pain, and other examples of inflammatory diseases include sepsis and chronic inflammation resulting from chronic viral or bacterial infections. In some cases, treatment of the condition may involve reducing the amount of at least one pro-inflammatory cytokine to a level that may be lower than that before treatment.

[0098] In some embodiments, the condition may be an infectious disease. For example, the infectious disease may be a bacterial infection, a viral infection, a fungal infection, or any combination thereof.

[0099] In some cases, infectious diseases may include infections caused by bacterial pathogens. Bacterial pathogens are not limited to the following groups, but include: Staphylococcus species, e.g., Staphylococcus aureus (e.g., Staphylococcus aureus NCTC 10442 and Staphylococcus aureus ATCC25923), Staphylococcus epidermidis; Chlamydia species, e.g., Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia psittaci; Enterococcus species, e.g., Enterococcus faecalis; Streptococcus pyogenes; Listeria species; Pseudomonas species; Mycobacterium species, e.g., Mycobacterium tuberculosis complex; Enterobacter species; Campylobacter species; Salmonella species; Streptococcus species, e.g., Streptococcus group A or B, Streptococcus pneumoniae; Helicobacter species, e.g., Helicobacter pylori, Helicobacter felis; Neisseria species, e.g., Neisseria gonorrhoea, Neisseria meningitidis; Borrelia burgdorferi; Shigella species, e.g., Shigella flexneri; Escherichia coli (E. coli 0157:H7 NCTC 12900); Haemophilus species, e.g., Haemophilus influenzae; Francisella tularensis; Bacillus species, e.g., Bacillus anthraces; Clostridia species, e.g., Clostridium botulinum, Clostridium difficile; Yersinia species, e.g., Yersinia pestis; Treponema species; Burkholderia species, e.g., Burkholderia cepacia complex, B. mallei, B. pseudomallei; Propionibacterium species, e.g., P.acnes, Acinetobacter species, Actinomyces species, Campylobacter species, Candida species, Corynebacterium minutissium, Corynebacterium pseudodiphtheriae, Corynebacterium stratium, Corynebacterium group G1, Corynebacterium group G2, Enterobacteriaceae, Enterococcus species, Klebsiella pneumoniae, Moraxella species, non-tuberculous mycobacteria species, Porphyromonas species, Prevotella melaninogenicus, Salmonella typhimurium, Serratia marcescens Streptococcus agalactiae, Staphylococcus salivarius, Streptococcus mitis, Streptococcus sanguis, Streptococcus pneumoniae, Vibrio It may originate from bacterial species selected from the group consisting of Cholerae, Coccidioides, or Cryptococcus species.

[0100] In some cases, infectious diseases may include viral infections. Viruses may originate from, but are not limited to, herpesviruses, poxviruses, hepadnaviruses, flaviviruses, togaviruses, coronaviruses, hepatitis C, hepatitis D, orthomyxoviruses, papillomaviruses, polyomaviruses, parvoviruses, cytomegaloviruses, Epstein-Barr viruses, smallpox viruses, bovinepox viruses, sheeppox viruses, Orff viruses, monkeypox viruses, vaccinia viruses, paramyxoviruses, retroviruses, adenoviruses, rhabdoviruses, bunyaviruses, filoviruses, alphaviruses, arenaviruses, lentiviruses, and any combination thereof. In some cases, viruses may be enveloped viruses. Examples of enveloped viruses include poxviruses, hepadnaviruses, flaviviruses, togaviruses, coronaviruses, hepatitis C, hepatitis D, orthomyxoviruses, cytomegaloviruses, Epstein-Barr virus, smallpox virus, bovine pox virus, sheeppox virus, Orff virus, monkeypox virus, vaccinia virus, rhabdovirus, bunyavirus, filovirus, alphavirus, arenavirus, and lentivirus.

[0101] In some cases, the infection may be caused by parasites selected from the group consisting of, but not limited to, Trypanosoma species (Trypanosoma cruzi, Trypansosoma brucei), Leishmania species, Giardia species, Trichomonas species, Entamoeba species, Naegleria species, Acanthanioeba species, Schistosoma species, Plasmodium species, Crytosporidium species, Isospora species, Balantidium species, Loa, Ascaris lumbricoides, Dirofilaria immitis, and Toxoplasma species, such as Toxoplasma gondii.

[0102] Fungal pathogens, limited species, including: *C. albicans* (genera Candida), *Epidermophyton*, *Exophiala*, *Microsporum*, *Trichophyton* (*T. rubrum*, *T. interdigitale*), *Tinea*, *Aspergillus*, *Blastomyces*, *Blastoschizomyces*, *Coccidioides*, and *Cryptococcus* (*Cryptococcus*). *Neoformans*, *Histoplasma*, *Paracoccidiomyces*, *Sporotrix*, *Absidia*, *Cladophialophora*, *Fonsecaea*, *Phialophora*, *Lacazia*, *Arthrographis*, *Acremoniwn*, *Actinomadura*, *Apophysomyces*, *Emmonsia*, *Basidiobolus*, *Beauveria*, *Chrysosporium*, *Conidiobolus*, *Cunninghamella*, *Fusarium*, *Geotrichum*, *Graphiwn*, *Leptosphaeria*, *Malassezia* (example: *Malassezia*) Furfur), Mucor species, Neotestudina species, Nocardia species, Nocardiopsis species, Paecilomyces species, Phona species, Piedraia species, Pneunwcystis species, Pseudallescheria species, Pyrenochaeta species, Rhizoinucor species, Rhizopus species, Rhodotorula species, Saccharomyces species, Scedosporium species, Scoplulariopsis species, Sporobolomyces species, S:yncephalastrum species, Trichoderma species, Trichosporon species, Ulocladium species, Ustilago species, Verticillium species.

[0103] The pharmaceutical compositions described herein may also be administered prophylactically to prevent the onset of any of the diseases or conditions described herein. For example, a pharmaceutical composition containing an antibiotic may be administered vaginally to a subject before surgery to prevent peritoneal infection. Medication / Pharmacokinetics

[0104] In some embodiments, pharmaceutical formulations may be formulated to optimize the pharmacokinetics / pharmacokinetics of the active ingredient or a salt thereof contained therein upon transvaginal administration of the pharmaceutical composition to a subject.

[0105] In some cases, a pharmaceutical composition comprising the activator or a salt thereof as described herein is available in doses of approximately 1 mg to approximately 1000 mg, approximately 5 mg to approximately 1000 mg, approximately 10 mg to approximately 1000 mg, approximately 15 mg to approximately 1000 mg, approximately 20 mg to approximately 1000 mg, approximately 25 mg to approximately 1000 mg, approximately 30 mg to approximately 1000 mg, approximately 35 mg to approximately 1000 mg, approximately 40 mg to approximately 1000 mg, approximately 45 mg to approximately 1000 mg, approximately 50 mg to approximately 1000 mg, approximately 55 mg to approximately 1000 mg, approximately 60 mg to approximately 1000 mg, approximately 65 mg to approximately 1000 mg, approximately 70 mg to approximately 1000 mg, approximately 75 mg to approximately 1000 mg, approximately 80 mg to approximately 1000 mg, approximately 85 mg to approximately 1000 mg, and approximately 90 mg to approximately 100 mg. 0mg, about 95mg to about 1000mg, about 100mg to about 1000mg, about 150mg to about 1000mg, about 200mg to about 1000mg, about 250mg to about 1000 mg, about 300mg to about 1000mg, about 350mg to about 1000mg, about 400mg to about 1000mg, about 450mg to about 1000mg, about 500mg to about 1000 It may be administered in doses of mg, approximately 550 mg to approximately 1000 mg, approximately 600 mg to approximately 1000 mg, approximately 650 mg to approximately 1000 mg, approximately 700 mg to approximately 1000 mg, approximately 750 mg to approximately 1000 mg, approximately 800 mg to approximately 1000 mg, approximately 850 mg to approximately 1000 mg, approximately 900 mg to approximately 1000 mg, or approximately 950 mg to approximately 1000 mg. In some cases, a pharmaceutical composition containing the activator or a salt thereof described herein may be administered in doses from 0.1 mg to a maximum of approximately 200 mg. In some cases, a pharmaceutical composition containing the activator or a salt thereof described herein may be administered in doses from 1 mg to a maximum of approximately 200 mg. In some cases, a pharmaceutical composition containing the activator or a salt thereof described herein may be administered in doses from approximately 1 mg to 100 mg. In some cases, pharmaceutical compositions containing the activators or salts thereof described herein may be administered in doses ranging from 1 mg to a maximum of about 100 mg.

[0106] In some cases, pharmaceutical compositions comprising the activators or salts thereof described herein are about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 4 4, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 9 8, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 1 39, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179180, 181, 182, 183, 184, 184, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 5 It may be administered in doses of 20, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, or 1000 mg. In some cases, the dose of the activator or its salt is less than 600 mg, less than 500 mg, less than 400 mg, less than 300 mg, less than 200 mg, or less than about 100 mg. In some cases, a pharmaceutical composition containing the activator or its salt described herein may be administered in a dose of about 100 mg. In some cases, a pharmaceutical composition containing the activator or its salt described herein may be administered in a dose of about 200 mg. In some cases, a pharmaceutical composition containing the activator or its salt described herein may be administered in a dose of about 300 mg. In some cases, a pharmaceutical composition containing the activator or its salt described herein may be administered in a dose of about 400 mg. In some cases, a pharmaceutical composition containing the activator or its salt described herein may be administered in a dose of about 500 mg.

[0107] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein will have a concentration of approximately 0.5 ng / mL to approximately 10 μg / mL, approximately 1 ng / mL to approximately 10 μg / mL, approximately 5 ng / mL to approximately 10 μg / mL, approximately 10 ng / mL to approximately 10 μg / mL, approximately 15 ng / mL to approximately 10 μg / mL, and approximately 20 μg / mL after about 1 minute to about 1, 2, 3, 4, 5, 6, 7 or 10 hours or more. ng / mL~about 10μg / mL, about 25ng / mL~about 10μg / mL, about 30ng / mL~about 10μg / mL, about 35ng / mL~about 10μg / mL, about 40ng / mL~about 10μg / m L, about 45ng / mL to about 10μg / mL, about 50ng / mL to about 10μg / mL, about 55ng / mL to about 10μg / mL, about 60ng / mL to about 10μg / mL, about 65ng / mL to about 1 0μg / mL, about 70ng / mL to about 10μg / mL, about 75ng / mL to about 10μg / mL, about 80ng / mL to about 10μg / mL, about 85ng / mL to about 10μg / mL, about 90ng / mL mL ~ approx. 10 μg / mL, approx. 95 ng / mL ~ approx. 10 μg / mL, approx. 100 ng / mL ~ approx. 10 μg / mL, approx. 200 ng / mL ~ approx. 10 μg / mL, approx. 300 ng / mL ~ approx. 10 μg / m An activator, its metabolite, or a salt thereof may be administered to provide plasma concentrations of approximately 400 ng / mL to approximately 10 μg / mL, approximately 500 ng / mL to approximately 10 μg / mL, approximately 600 ng / mL to approximately 10 μg / mL, approximately 700 ng / mL to approximately 10 μg / mL, approximately 800 ng / mL to approximately 10 μg / mL, approximately 900 ng / mL to approximately 10 μg / mL, or approximately 1 μg / mL to approximately 10 μg / mL. In some cases, a pharmaceutical composition containing an activator or a salt thereof as described herein may be administered to provide plasma concentrations of the activator, its metabolite, or a salt thereof of approximately 0.5 ng / mL to approximately 10 ng / mL.

[0108] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein will, after administration to a subject, reach at least about 200 ng / mL, 195 ng / mL, 190 ng / mL, 185 ng / mL, 180 ng / mL, 175 ng / mL, 170 ng / mL, 165 ng / mL, 160 ng / mL, 155 ng / mL, 150 ng / mL, 145 ng / mL, 140 ng / mL, 135 ng / mL, 130 ng / mL, 125 ng / mL, An activator, its metabolite, or a salt thereof may be administered to provide plasma concentrations of 120 ng / mL, 115 ng / mL, 110 ng / mL, 105 ng / mL, 100 ng / mL, 95 ng / mL, 90 ng / mL, 85 ng / mL, 80 ng / mL, 75 ng / mL, 70 ng / mL, 65 ng / mL, 60 ng / mL, 55 ng / mL, 50 ng / mL, 45 ng / mL, 40 ng / mL, 35 ng / mL, 30 ng / mL, 25 ng / mL, 20 ng / mL, 15 ng / mL, 10 ng / mL, or 5 ng / mL.

[0109] In some embodiments, vaginal administration of the compositions provided herein reduces the systemic concentration of the activator or its salt in the subject by at least 10 times compared to oral administration of the activator or its salt in the subject. In some embodiments, vaginal administration of the compositions provided herein reduces the systemic concentration of the activator or its salt in the subject by at least 20 times compared to oral administration of the activator or its salt in the subject. In some embodiments, vaginal administration of the compositions provided herein reduces the systemic concentration of the activator or its salt in the subject by at least 30 times compared to oral administration of the activator or its salt in the subject. In some embodiments, vaginal administration of the compositions provided herein reduces the systemic concentration of the activator or its salt in the subject by at least 40 times compared to oral administration of the activator or its salt in the subject. In some embodiments, vaginal administration of the compositions provided herein containing about 100 mg of the activator or its salt results in serum concentrations ranging from about 1 ng / mL to about 50 ng / mL. In some embodiments, vaginal administration of a composition provided herein containing about 100 mg of the activator or a salt thereof reaches serum concentrations of about 0.1 ng / mL to 50 ng / mL. In some embodiments, vaginal administration of a composition provided herein containing about 100 mg of the activator or a salt thereof reaches serum concentrations of about 1 ng / mL to about 10 ng / mL. In some embodiments, a pharmaceutical composition containing 600 mg of the activator or a salt thereof, when administered orally to a subject, reaches serum concentrations of about 100 ng / mL to about 500 ng / mL.

[0110] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein may result in at least approximately 200 ng / mL, 195 ng / mL, 190 ng / mL, 185 ng / mL, 180 ng / mL, 175 ng / mL, 170 ng / mL, 165 ng / mL, 160 ng / mL, 155 ng / mL, 150 ng / mL, 145 ng / mL, 140 ng / mL, 135 ng / mL, 130 ng / mL, 125 ng / mL, 120 ng / mL, 115 ng / mL, 110 ng / mL, 105 ng / mL, 100 ng / mL, 95 ng / mL, 90 ng / mL, 85 ng / mL in the ascites fluid after administration to a subject, approximately 1 minute to approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours later. Activators, their metabolites, or salts thereof may be administered to provide concentrations of mL, 80 ng / mL, 75 ng / mL, 70 ng / mL, 65 ng / mL, 60 ng / mL, 55 ng / mL, 50 ng / mL, 45 ng / mL, 40 ng / mL, 35 ng / mL, 30 ng / mL, 25 ng / mL, 20 ng / mL, 15 ng / mL, 10 ng / mL, 9 ng / mL, 8 ng / mL, 7 ng / mL, 6 ng / mL, 5 ng / mL, 4 ng / mL, 3 ng / mL, 2 ng / mL, 1 ng / mL, 0.9 ng / mL, 0.8 ng / mL, 0.7 ng / mL, 0.6 ng / mL, 0.5 ng / mL, 0.4 ng / mL, 0.3 ng / mL, 0.2 ng / mL, or 0.1 ng / mL.

[0111] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein, in the ascites fluid after administration to a subject, shows concentrations of approximately 0.5 ng / mL to approximately 100 ng / mL, approximately 0.5 ng / mL to approximately 90 ng / mL, approximately 0.5 ng / mL to approximately 80 ng / mL, approximately 0.5 ng / mL to approximately 70 ng / mL, approximately 0.5 ng / mL to approximately 60 ng / mL, approximately 0.5 ng / mL to approximately 50 ng / mL, approximately 0.5 ng / mL to approximately 40 ng / mL, and approximately 0.5 ng / mL to approximately 30 ng / mL after approximately 1 minute to approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours. An activator, its metabolite or a salt thereof may be administered to provide concentrations of approximately 0.5 ng / mL to approximately 20 ng / mL, approximately 0.5 ng / mL to approximately 10 ng / mL, approximately 0.5 ng / mL to approximately 9 ng / mL, approximately 0.5 ng / mL to approximately 8 ng / mL, approximately 0.5 ng / mL to approximately 7 ng / mL, approximately 0.5 ng / mL to approximately 6 ng / mL, approximately 0.5 ng / mL to approximately 5 ng / mL, approximately 0.5 ng / mL to approximately 4 ng / mL, approximately 0.5 ng / mL to approximately 3 ng / mL, approximately 0.5 ng / mL to approximately 2 ng / mL, or approximately 0.5 ng / mL to approximately 1 ng / mL.

[0112] In some cases, a pharmaceutical composition containing the activator or a salt thereof described herein will produce at least approximately 200 ng / mg, 195 ng / mg, 190 ng / mg, 185 ng / mg, 180 ng / mg, 175 ng / mg, 170 ng / mg, 165 ng / mg, 160 ng / mg, 155 ng / mg, 150 ng / mg, 145 ng / mg, 140 ng / mg, 135 ng / mg, 130 ng / mg, 125 ng / mg, 120 ng / mg, 115 ng / mg, 110 ng / mg, 105 ng / mg, 100 ng / mg, 95 ng / mg, 90 ng / mg, and 85 ng in the peritoneal tissue after administration to a subject, approximately 1 minute to approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours later. The activator, its metabolites or salts thereof may be administered to provide concentrations of 0.1 ng / mg, 80 ng / mg, 75 ng / mg, 70 ng / mg, 65 ng / mg, 60 ng / mg, 55 ng / mg, 50 ng / mg, 45 ng / mg, 40 ng / mg, 35 ng / mg, 30 ng / mg, 25 ng / mg, 20 ng / mg, 15 ng / mg, 10 ng / mg, 9 ng / mg, 8 ng / mg, 7 ng / mg, 6 ng / mg, 5 ng / mg, 4 ng / mg, 3 ng / mg, 2 ng / mg, 1 ng / mg, 0.9 ng / mg, 0.8 ng / mg, 0.7 ng / mg, 0.6 ng / mg, 0.5 ng / mg, 0.4 ng / mg, 0.3 ng / mg, 0.2 ng / mg, or 0.1 ng / mg.

[0113] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein in the peritoneal tissue after administration to a subject yields approximately 0.5 ng / mg to approximately 100 ng / mg, approximately 0.5 ng / mg to approximately 90 ng / mg, approximately 0.5 ng / mg to approximately 80 ng / mg, approximately 0.5 ng / mg to approximately 70 ng / mg, approximately 0.5 ng / mg to approximately 60 ng / mg, approximately 0.5 ng / mg to approximately 50 ng / mg, approximately 0.5 ng / mg to approximately 40 ng / mg, and approximately 0.5 ng / mg to approximately 30 ng / mg at approximately 1 minute to approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours later. The activator, its metabolites or salts thereof may be administered to provide concentrations of mg, approximately 0.5 ng / mg to approximately 20 ng / mg, approximately 0.5 ng / mg to approximately 10 ng / mg, approximately 0.5 ng / mg to approximately 9 ng / mg, approximately 0.5 ng / mg to approximately 8 ng / mg, approximately 0.5 ng / mg to approximately 7 ng / mg, approximately 0.5 ng / mg to approximately 6 ng / mg, approximately 0.5 ng / mg to approximately 5 ng / mg, approximately 0.5 ng / mg to approximately 4 ng / mg, approximately 0.5 ng / mg to approximately 3 ng / mg, approximately 0.5 ng / mg to approximately 2 ng / mg, or approximately 0.5 ng / mg to approximately 1 ng / mg.

[0114] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein, after administration to a subject, lasts for approximately 1 minute to 600 minutes, approximately 1 minute to 590 minutes, approximately 1 minute to 580 minutes, approximately 1 minute to 570 minutes, approximately 1 minute to 560 minutes, approximately 1 minute to 550 minutes, approximately 1 minute to 540 minutes, approximately 1 minute to 530 minutes, approximately 1 minute to 520 minutes, approximately 1 minute to 510 minutes, approximately 1 minute to 500 minutes, approximately 1 minute to 490 minutes, approximately 1 minute to 480 minutes, and approximately 1 minute to 470 minutes. Between approximately 1 minute and 460 minutes, approximately 1 minute and 450 minutes, approximately 1 minute and 440 minutes, approximately 1 minute and 430 minutes, approximately 1 minute and 420 minutes, approximately 1 minute and 410 minutes, approximately 1 minute and 400 minutes, approximately 1 minute and 390 minutes, approximately 1 minute and 380 minutes, approximately 1 minute and 370 minutes, approximately 1 minute and 360 minutes, approximately 1 minute and 350 minutes, approximately 1 minute and 340 minutes, approximately 1 minute and 330 minutes, approximately 1 minute and 320 minutes, approximately 1 minute and 310 minutes, approximately 1 minute and 300 minutes, approximately 1 minute and 290 minutes, approximately 1 minute to approximately 280 minutes, approximately 1 minute to approximately 270 minutes, approximately 1 minute to approximately 260 minutes, approximately 1 minute to approximately 250 minutes, approximately 1 minute to approximately 240 minutes, approximately 1 minute to approximately 230 minutes, approximately 1 minute to approximately 220 minutes, approximately 1 minute to approximately 210 minutes, approximately 1 minute to approximately 200 minutes, approximately 1 minute to approximately 190 minutes, approximately 1 minute to approximately 180 minutes, approximately 1 minute to approximately 170 minutes, approximately 1 minute to approximately 160 minutes, approximately 1 minute to approximately 150 minutes, approximately 1 minute to approximately 140 minutes, approximately 1 minute to approximately 130 minutes, approximately 1 minute to approximately 120 minutes, approximately 1 minute to approximately 110 minutes, approximately 1 minute It may be administered to provide an activator or a salt thereof with a Tmax of approximately 1 to 100 minutes, approximately 1 minute to approximately 90 minutes, approximately 1 minute to approximately 80 minutes, approximately 1 minute to approximately 70 minutes, approximately 1 minute to approximately 60 minutes, approximately 1 minute to approximately 50 minutes, approximately 1 minute to approximately 40 minutes, approximately 1 minute to approximately 30 minutes, approximately 1 minute to approximately 20 minutes, approximately 1 minute to approximately 10 minutes, approximately 1 minute to approximately 9 minutes, approximately 1 minute to approximately 8 minutes, approximately 1 minute to approximately 7 minutes, approximately 1 minute to approximately 6 minutes, approximately 1 minute to approximately 5 minutes, approximately 1 minute to approximately 4 minutes, approximately 1 minute to approximately 3 minutes, or approximately 1 minute to approximately 2 minutes.

[0115] In some cases, a pharmaceutical composition comprising the activator or a salt thereof described herein, after administration to a subject, contains at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 9 5, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179 The activator, its metabolites or salts thereof may be administered to provide Tmax for 180, 181, 182, 183, 184, 184, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, or 200 minutes.In some cases, a pharmaceutical composition comprising the activators or salts described herein will, after administration to a subject, yield at least about 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7 An activator, its metabolite, or a salt thereof may be administered to provide a Tmax of 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0 hours.

[0116] In some cases, a pharmaceutical composition comprising the activators or salts thereof described herein may be administered to a subject to provide, after administration, at least about 1,000 μg / mL, 950 μg / mL, 900 μg / mL, 850 μg / mL, 800 μg / mL, 750 μg / mL, 700 μg / mL, 650 μg / mL, 600 μg / mL, 550 μg / mL, 500 μg / mL, 450 μg / mL, 400 μg / mL, 350 μg / mL, 300 μg / mL, 250 μg / mL, 200 μg / mL, 150 μg / mL, 100 μg / mL, or 50 μg / mL of the activator, its metabolites or salts thereof. In some cases, a pharmaceutical composition comprising the activators or salts thereof described herein may be administered to a subject to provide, after administration, at least about 100 μg / mL, 95 μg / mL, 90 μg / mL, 85 μg / mL, 80 μg / mL, 75 μg / mL, 70 μg / mL, 65 μg / mL, 60 μg / mL, 55 μg / mL, 50 μg / mL, 45 μg / mL, 40 μg / mL, 35 μg / mL, 30 μg / mL, 25 μg / mL, 20 μg / mL, 15 μg / mL, 10 μg / mL, 5 μg / mL, 4 μg / mL, 3 μg / mL, 2 μg / mL, or 1 μg / mL of the activator, its metabolites or salts thereof. In some cases, the activators described herein, their salts, or pharmaceutical compositions containing the activators or their salts may be administered to a subject to provide, after administration, at least about 1,000 ng / mL, 950 ng / mL, 900 ng / mL, 850 ng / mL, 800 ng / mL, 750 ng / mL, 700 ng / mL, 650 ng / mL, 600 ng / mL, 550 ng / mL, 500 ng / mL, 450 ng / mL, 400 ng / mL, 350 ng / mL, 300 ng / mL, 250 ng / mL, 200 ng / mL, 150 ng / mL, 100 ng / mL, or 50 ng / mL of the activator, its metabolites, or salts thereof with a Cmax.In some cases, a pharmaceutical composition comprising the activators or salts thereof described herein may be administered to a subject to provide, after administration, at least about 100 ng / mL, 95 ng / mL, 90 ng / mL, 85 ng / mL, 80 ng / mL, 75 ng / mL, 70 ng / mL, 65 ng / mL, 60 ng / mL, 55 ng / mL, 50 ng / mL, 45 ng / mL, 40 ng / mL, 35 ng / mL, 30 ng / mL, 25 ng / mL, 20 ng / mL, 15 ng / mL, 10 ng / mL, or 5 ng / mL of the activator, its metabolites or salts thereof. In some cases, a pharmaceutical composition containing the activator or a salt thereof described herein contains at least about 50 ng / mL, 49 ng / mL, 48 ng / mL, 47 ng / mL, 46 ng / mL, 45 ng / mL, 44 ng / mL, 43 ng / mL, 42 ng / mL, 41 ng / mL, 40 ng / mL, 39 ng / mL, 38 ng / mL, 37 ng / mL, 36 ng / mL, 35 ng / mL, 34 ng / mL, 33 ng / mL, 32 ng / mL, 31 ng / mL, 30 ng / mL, 29 ng / mL, 28 ng / mL, 27 ng / mL, and 26 ng An activator with a Cmax of 0.5 ng / mL, 25 ng / mL, 24 ng / mL, 23 ng / mL, 22 ng / mL, 21 ng / mL, 20 ng / mL, 19 ng / mL, 18 ng / mL, 17 ng / mL, 16 ng / mL, 15 ng / mL, 14 ng / mL, 13 ng / mL, 12 ng / mL, 11 ng / mL, 10 ng / mL, 9 ng / mL, 8 ng / mL, 7 ng / mL, 6 ng / mL, 5 ng / mL, 4 ng / mL, 3 ng / mL, 2 ng / mL, 1 ng / mL, or 0.5 ng / mL may be administered to provide the activator, its metabolites, or salts thereof.

[0117] In some embodiments, a pharmaceutical composition comprising the activator or a salt thereof described herein, after administration to a subject, contains at least about 10,000 ng*h / mL, 9,900 ng*h / mL, 9,800 ng*h / mL, 9,700 ng*h / mL, 9,600 ng*h / mL, 9,500 ng*h / mL, 9,400 ng*h / mL, 9,300 ng*h / mL, 9,200 ng*h / mL, 9,100 ng*h / mL, 9,000 ng*h / mL, 8,900ng*h / mL, 8,800ng*h / mL, 8,700ng*h / mL, 8,600ng*h / mL, 8,500ng*h / mL, 8,400ng*h / mL, 8,300ng*h / mL, 8,200ng*h / m L, 8,100ng*h / mL, 8,000ng*h / mL, 7,900ng*h / mL, 7,800ng*h / mL, 7,700ng*h / mL, 7,600ng*h / mL, 7,500ng*h / mL, 7,400ng*h / mL, 7,300ng*h / mL, 7,200ng*h / mL, 7,100ng*h / mL, 7,000ng*h / mL, 6,900ng*h / mL, 6,800ng*h / mL, 6,700ng*h / mL, 6,600ng* h / mL, 6,500ng*h / mL, 6,400ng*h / mL, 6,300ng*h / mL, 6,200ng*h / mL, 6,100ng*h / mL, 6,000ng*h / mL, 5,900ng*h / mL, 5,800ng *h / mL, 5,700ng*h / mL, 5,600ng*h / mL, 5,500ng*h / mL, 5,400ng*h / mL, 5,300ng*h / mL, 5,200ng*h / mL, 5,100ng*h / mL, 5,000 ng*h / mL, 4,500ng*h / mL, 4,000ng*h / mL, 3,500ng*h / mL, 3,000ng*h / mL, 2,500ng*h / mL, 2,000ng*h / mL, 1,500ng*h / mL or 1,An activator, its metabolite or a salt thereof, with an AUC(0-t) of 900 ng*h / mL may be administered to provide at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 3 after administration of a pharmaceutical composition containing the activator or a salt thereof. It could be 5, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 or 90 hours.

[0118] In some embodiments, a pharmaceutical composition comprising the activator or a salt thereof described herein, after administration to a subject, contains at least about 10,000 ng*h / mL, 9,900 ng*h / mL, 9,800 ng*h / mL, 9,700 ng*h / mL, 9,600 ng*h / mL, 9,500 ng*h / mL, 9,400 ng*h / mL, 9,300 ng*h / mL, 9,200 ng*h / mL, 9,100 ng*h / mL, 9,000 ng*h / mL, 8,900ng*h / mL, 8,800ng*h / mL, 8,700ng*h / mL, 8,600ng*h / mL, 8,500ng*h / mL, 8,400ng*h / mL, 8,300ng*h / mL, 8,200ng*h / m L, 8,100ng*h / mL, 8,000ng*h / mL, 7,900ng*h / mL, 7,800ng*h / mL, 7,700ng*h / mL, 7,600ng*h / mL, 7,500ng*h / mL, 7,400ng*h / mL, 7,300ng*h / mL, 7,200ng*h / mL, 7,100ng*h / mL, 7,000ng*h / mL, 6,900ng*h / mL, 6,800ng*h / mL, 6,700ng*h / mL, 6,600ng* h / mL, 6,500ng*h / mL, 6,400ng*h / mL, 6,300ng*h / mL, 6,200ng*h / mL, 6,100ng*h / mL, 6,000ng*h / mL, 5,900ng*h / mL, 5,800ng *h / mL, 5,700ng*h / mL, 5,600ng*h / mL, 5,500ng*h / mL, 5,400ng*h / mL, 5,300ng*h / mL, 5,200ng*h / mL, 5,100ng*h / mL, 5,000 ng*h / mL, 4,500ng*h / mL, 4,000ng*h / mL, 3,500ng*h / mL, 3,000ng*h / mL, 2,500ng*h / mL, 2,000ng*h / mL, 1,500ng*h / mL or 1,An activator, its metabolite, or a salt thereof may be administered to provide an AUC(0-t) of 900 ng*h / mL, where t is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 after administration of the pharmaceutical composition containing the activator or a salt thereof. , could be 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 or 90 days.

[0119] In some exemplary embodiments, a pharmaceutical composition comprising the activator or a salt thereof described herein, after administration to a subject, results in a blood glucose level of approximately 1,000 ng*h / mL to approximately 10,000 ng*h / mL, approximately 1,000 ng*h / mL to approximately 9,000 ng*h / mL, approximately 1,000 ng*h / mL to approximately 8,000 ng*h / mL, and approximately 1,000 ng*h / mL to approximately 7,000 Activators with AUC(0-t) of ng*h / mL, approximately 1,000ng*h / mL to approximately 6,000ng*h / mL, approximately 1,000ng*h / mL to approximately 5,000ng*h / mL, approximately 1,000ng*h / mL to approximately 4,000ng*h / mL, approximately 1,000ng*h / mL to approximately 3,000ng*h / mL, or approximately 1,000ng*h / mL to approximately 2,000ng*h / mL. It may be administered to provide its metabolite or a salt thereof, and t is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 after administration of a pharmaceutical composition containing the activator or a salt thereof. , 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 or 90 days.

[0120] In some exemplary embodiments, when the pharmaceutical formulation is administered to primates, the activator or a salt thereof is released within about 1 minute to about 1 hour. max C for approximately 1 minute to approximately 8 hours max AUC of approximately 0.1 μg.hr / L to approximately 1,000 μg.hr / L 0>24 hour They can be produced to have a half-life of approximately 2 hours to approximately 24 hours, or a combination thereof.

[0121] In some embodiments, the pharmaceutical formulation may be produced such that, when the pharmaceutical formulation is administered to a subject, the activator or a salt thereof can be substantially localized to the peritoneal organs or tissues of the subject. Examples of organs or tissues of the peritoneal cavity may be those depicted in Figure 1. The peritoneal cavity may contain ascites. Peritoneal organs or tissues may include, but are not limited to, the bladder, gallbladder, intestines, uterus, endometrium, myometrium, epithelium, stomach, ovaries, ovarian cortex, ovarian epithelium, liver, spleen, or kidneys.

[0122] In some embodiments, when a pharmaceutical formulation is administered to a subject, the activator is approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45 ,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98,9 9, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 13 9, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179 It may have a half-life of 180, 181, 182, 183, 184, 184, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, or 200 minutes.In some embodiments, when a pharmaceutical formulation is administered to a subject, the activator or a salt thereof is approximately 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7 It may have a half-life of 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0 hours.In some embodiments, when a pharmaceutical formulation is administered to a subject, the activator or a salt thereof is administered for approximately 1 minute to approximately 600 minutes, approximately 1 minute to approximately 590 minutes, approximately 1 minute to approximately 580 minutes, approximately 1 minute to approximately 570 minutes, approximately 1 minute to approximately 560 minutes, approximately 1 minute to approximately 550 minutes, approximately 1 minute to approximately 540 minutes, approximately 1 minute to approximately 530 minutes, approximately 1 minute to approximately 520 minutes, approximately 1 minute to approximately 510 minutes, approximately 1 minute to approximately 500 minutes, approximately 1 minute to approximately 490 minutes, approximately 1 minute to approximately 480 minutes, approximately 1 minute to approximately 470 minutes, and approximately 1 1 minute to approximately 460 minutes, approximately 1 minute to approximately 450 minutes, approximately 1 minute to approximately 440 minutes, approximately 1 minute to approximately 430 minutes, approximately 1 minute to approximately 420 minutes, approximately 1 minute to approximately 410 minutes, approximately 1 minute to approximately 400 minutes, approximately 1 minute to approximately 390 minutes, approximately 1 minute to approximately 380 minutes, approximately 1 minute to approximately 370 minutes, approximately 1 minute to approximately 360 minutes, approximately 1 minute to approximately 350 minutes, approximately 1 minute to approximately 340 minutes, approximately 1 minute to approximately 330 minutes, approximately 1 minute to approximately 320 minutes, approximately 1 minute to approximately 310 minutes, approximately 1 minute to approximately 300 minutes, approximately 1 minute to approximately 290 minutes, approximately 1 minute to approximately 280 minutes, approximately 1 minute to approximately 270 minutes, approximately 1 minute to approximately 260 minutes, approximately 1 minute to approximately 250 minutes, approximately 1 minute to approximately 240 minutes, approximately 1 minute to approximately 230 minutes, approximately 1 minute to approximately 220 minutes, approximately 1 minute to approximately 210 minutes, approximately 1 minute to approximately 200 minutes, approximately 1 minute to approximately 190 minutes, approximately 1 minute to approximately 180 minutes, approximately 1 minute to approximately 170 minutes, approximately 1 minute to approximately 160 minutes, approximately 1 minute to approximately 150 minutes, approximately 1 minute to approximately 140 minutes, approximately 1 minute to approximately 130 minutes, approximately 1 minute to approximately 120 minutes, approximately 1 minute It may have a half-life of approximately 1 to 110 minutes, approximately 1 minute to 100 minutes, approximately 1 minute to 90 minutes, approximately 1 minute to 80 minutes, approximately 1 minute to 70 minutes, approximately 1 minute to 60 minutes, approximately 1 minute to 50 minutes, approximately 1 minute to 40 minutes, approximately 1 minute to 30 minutes, approximately 1 minute to 20 minutes, approximately 1 minute to 10 minutes, approximately 1 minute to 9 minutes, approximately 1 minute to 8 minutes, approximately 1 minute to 7 minutes, approximately 1 minute to 6 minutes, approximately 1 minute to 5 minutes, approximately 1 minute to 4 minutes, approximately 1 minute to 3 minutes, or approximately 1 minute to 2 minutes.

[0123] Methods for detecting drugs A method for detecting an active agent, its metabolite, or a salt thereof in a sample derived from a subject after administration of a pharmaceutical composition to the subject is provided herein. The sample derived from the subject can be blood or any excreted fluid. Non-limiting examples can include saliva, blood, serum, cerebrospinal fluid, semen, feces, plasma or urine.

[0124] In some exemplary embodiments, the method can be a method for detecting danazol or a salt thereof in a sample derived from a subject. The method can include contacting a portion of the sample derived from the subject with albumin; and detecting danazol or a salt thereof by mass spectrometry. The albumin can include human or bovine albumin. In some cases, the albumin can be serum albumin.

[0125] Detection of an active agent such as danazol can be performed by contacting the active agent with albumin to form an albumin adduct and using mass spectrometry to determine the presence of the adduct. The method can further include comparison with an internal standard. Exemplary internal standards can include 19-norethindrone.

[0126] The mass spectrometer can include a single or tandem mass spectrometer. The mass spectrometer can include an electrospray ionization device, a matrix-assisted laser desorption / ionization ionization device, an electron ionization device, a fast atom bombardment ionization device or a chemical ionization device.

[0127] An exemplary method can include detecting danazol by determining the amount of [M+H] + ions. In some cases, the [M+H] + ions can include an m / z of about 338.

[0128] In some embodiments, the sample may be dried after contact with albumin. In some embodiments, the sample may be reconstituted, resuspended, or diluted with a resuspension buffer after contact with albumin. The resuspension buffer may be a buffer such as saline, citrate, phosphate, phosphate-buffered saline, acetate, glycine, tris(hydroxymethyl)aminomethane (Tris) hydrochloride, Tris-buffered saline (TBS), 3-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]propan-1-sulfonic acid (TAPS), bicine, tricine, 3-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]-2-hydroxypropan-1-sulfonic acid It may be ethanesulfonic acid (TAPSO), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), 3-(N-morpholino)propanesulfonic acid (MOPS), 2-(N-morpholino)ethanesulfonic acid (MES), 2-[[1,3-dihydroxy-2-(hydroxymethyl)propane-2-yl]amino]ethanesulfonic acid (TES), cacodylate, glycine, carbonate, or any combination thereof.

[0129] kit A kit is disclosed herein. In some embodiments, the composition may be packaged in a container. In some embodiments, the kit may further include instructions for administering a unit dose of the composition to a subject.

[0130] The method for preparing the kit may include placing the pharmaceutical composition into a container. The method may further include including instructions for use. In some cases, the instructions for use may instruct the administration of a unit dose of the composition to a subject.

[0131] A kit for determining the amount of danazol in a sample is provided herein. The kit may include a sample collection container; albumin; and instructions for use. In some cases, the internal standard may be 19-norethindrone or a salt thereof. The instructions for use may instruct the user to collect the sample in the sample collection container; to contact a portion of the sample with a predetermined amount of albumin; and to detect danazol or a salt thereof by mass spectrometry. In some cases, the kit may further include an internal standard. The internal standard may be 19-norethindrone or a salt thereof.

[0132] Treatment method Provided herein are methods for treating a disease of a subject, comprising administering a composition provided herein to the subject. In some embodiments, the composition comprises danazol or a salt thereof. In some embodiments, the subject has, is suspected of having, has been diagnosed with, or is at risk of developing, endometrial disease, infection, cancer, dysmenorrhea, dyspareunia, pelvic pain, rectovaginal plaque, infertility, premenopausal, menopausal, or hormonal imbalance.

[0133] Furthermore, provided herein are methods for treating endometrial disease or disorder in a subject requiring treatment of such disease or disorder, comprising administering a composition provided herein to the subject requiring treatment. In some embodiments, the subject has a menstrual cycle. In some embodiments, the subject has an endometrial lesion. In some embodiments, the subject had one or more occurrences of infertility or miscarriage.

[0134] Endometriosis is a condition in which endometrial tissue is found outside the uterus. It can be "trapped" in the pelvic region and abdomen, and rarely in other parts of the body. Symptoms of endometriosis include, for example, menstrual cramps, painful intercourse, pelvic pain, reduced fertility, painful bowel movements, painful urination, and blood in the urine or stool. In rare cases, endometriotic lesions can develop in other parts of the body. This can cause unusual pain in parts of the body that occur simultaneously with menstrual cramps.

[0135] Furthermore, provided herein are methods for treating a disease of a subject, comprising administering a composition provided herein to the subject, wherein the composition comprises two or more agents provided herein. In some embodiments, the composition comprises (1) danazol, its derivatives or salts; and (2) estrogen, its derivatives or salts. Exemplary Embodiments

[0136] Provided herein are compositions comprising a cream, polybioadhesive gel, or liquid emulsion.

[0137] Provided herein is a cream comprising: (a) a finely powdered danazol or a salt thereof present in an amount of 5.0% to about 7.0% by weight-to-volume (w / v) of the cream; (b) a polyglycolized oleic acid glyceride or a mixture thereof present in an amount of 10% to 20% (w / v) of the cream; (c) a cellulose polymer present in an amount of 1.0% to 3.0% (w / v) of the cream; and (d) one or more water-insoluble bioadhesives, wherein at least one water-insoluble bioadhesive is polycarbophil present in an amount of 1.0% to about 2.0% (w / v) of the cream, and the cream comprises at least 60% (w / v) water and one or more components selected from carbomer, methylparaben, sorbic acid, poloxamer, and sodium carboxymethylcellulose. Furthermore, this specification provides a cream containing carbomer, wherein the carbomer is present in an amount of 0.1% to 2.0% (w / v) of the cream. Furthermore, this specification provides a cream in which a water-insoluble bioadhesive substance is dispersed or suspended in the aqueous phase of the cream. Furthermore, this specification provides a cream in the form of a liquid emulsion. Furthermore, this specification provides a cream containing at least 70% water. Furthermore, this specification provides a cream containing sorbic acid, wherein the sorbic acid is present in an amount of 0.1% to 2.0% (w / v) of the cream. Furthermore, this specification provides a cream in which micronized danazol or a salt thereof is dispersed or suspended in the oily phase of the cream. Furthermore, this specification provides a cream in which the dose of micronized danazol or a salt thereof is between 50 milligrams and 200 milligrams. Furthermore, this specification provides a cream formulated for vaginal administration. Furthermore, provided herein are creams in which, upon vaginal administration, the activator or a salt thereof diffuses into the ascites fluid, and further reaches the pelvic tissue via transvaginal absorption and countercurrent movement between the uterine lymphatic vessels or veins and arteries, thereby localizing the activator into the peritoneal cavity.

[0138] Provided herein are compositions comprising a polybioadhesive gel, including: (a) an activator or a salt thereof; (b) one or more water-insoluble bioadhesive materials; (c) at least one oleogel; and (d) at least one aqueous gel, which, upon administration to a subject, allows the activator to localize in peritoneal tissue and induce pelvic transit metabolism in the subject. Further provided herein are compositions in which the activator is a synthetic steroid. Further provided herein are compositions in which the synthetic steroid is danazol or a salt thereof. Further provided herein are compositions in which the polybioadhesive gel is formulated as a liquid emulsion. Further provided herein are compositions formulated for vaginal administration. Further provided herein are compositions in which, upon vaginal administration of the composition, the activator or a salt thereof diffuses into the ascites fluid and further reaches the pelvic tissue via transvaginal absorption and countercurrent movement between the uterine lymphatic vessels or veins and arteries, localizing the activator in the peritoneal cavity.

[0139] Provided herein are compositions comprising the components of Table 3 and the respective amounts (w / v) of each component. Further provided herein are compositions formulated for vaginal administration. Further provided herein are compositions in which, upon vaginal administration, the activator or a salt thereof diffuses into the ascites fluid, localizing the activator in the peritoneal cavity, and further reaching the pelvic tissue via transvaginal absorption and countercurrent movement between the uterine and vaginal lymphatic vessels or veins and arteries.

[0140] Provided herein is a method for treating a disease or condition of the endometrium of a subject, comprising administering a polybioadhesive gel to the subject via vaginal administration, wherein the polybioadhesive gel comprises (a) an activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel, wherein the vaginal administration of the polybioadhesive gel localizes the activator to the peritoneal tissue, thereby treating the disease or condition of the endometrium. Further provided herein is a method by which the vaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or a salt thereof compared to an equivalent oral composition containing the same activator. Further provided herein is a method in which the vaginal administration is performed once every 24 hours. Further provided herein is a method in which the activator is a synthetic steroid. Further provided herein is a method in which the synthetic steroid is danazol or a salt thereof. Further provided herein is a method in which the activator is micronized. Furthermore, this specification provides a method for a subject having or suspected infertility. Furthermore, this specification provides a method for a subject having or suspected endometriosis. Furthermore, this specification provides a method by which vaginal administration reduces the number of endometrial lesions in a subject by at least 10% compared to the number of endometrial lesions in the subject before vaginal administration of the polybioadhesive gel. Furthermore, this specification provides a method by which vaginal administration reduces the size of one or more endometrial lesions in a subject by at least 10% compared to the size of one or more endometrial lesions before vaginal administration of the polybioadhesive gel. Furthermore, this specification provides a method by which the polybioadhesive gel is formulated as a liquid emulsion. Furthermore, this specification provides a method by which the endometrial disorder is endometriosis, adenomyosis, or a combination thereof. Furthermore, this specification provides a method by which the polybioadhesive gel does not destroy the ovarian function of the subject. Furthermore, this specification provides a method by which a polybioadhesive gel reduces the subject's pain by at least 10% compared to the pain identified by the subject before the polybioadhesive gel was administered vaginally.A method is further provided herein for administering a composition provided herein vaginally to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 10 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein vaginally to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 15 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein vaginally to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 20 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein vaginally to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 25 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein vaginally to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 30 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 35 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 40 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 45 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 50 times compared to oral administration of the activator or a salt thereof to a subject.

[0141] Provided herein is a method for inducing pelvic transit metabolism of an activator in a subject, comprising: (a) the activator or a salt thereof; (b) one or more water-insoluble bioadhesive substances; (c) at least one oleogel; and (d) at least one aqueous gel, wherein the vaginal administration increases the level of the activator in the peritoneal tissue relative to the level of the activator in the subject's serum, and maintains the ratio of the level of the activator in the subject's ascites to the level of the activator in the subject's ascites, which is substantially equivalent to the oral administration of an oral pharmaceutical composition containing a substantially equivalent dose of the activator or a salt thereof, thereby localizing the activator to the peritoneal tissue and inducing pelvic transit metabolism in the subject. Further provided herein is a method in which the vaginal administration is performed once every 24 hours. Further provided herein is a method in which the activator is a synthetic steroid. Further provided herein is a method in which the synthetic steroid is danazol or a salt thereof. Furthermore, this specification provides a method for micronizing an activator. Furthermore, this specification provides a method for a subject having or suspected infertility. Furthermore, this specification provides a method for a subject having or suspected endometriosis. Furthermore, this specification provides a method for vaginal administration that reduces the number of endometrial lesions in a subject by at least 10% compared to the number of endometrial lesions in the subject before vaginal administration of the polybioadhesive gel. Furthermore, this specification provides a method for vaginal administration that reduces the size of one or more endometrial lesions in a subject by at least 10% compared to the size of one or more endometrial lesions before vaginal administration of the polybioadhesive gel. Furthermore, this specification provides a method for formulating the polybioadhesive gel as a liquid emulsion. Furthermore, this specification provides a method for endometrial disorders being endometriosis, adenomyosis, or a combination thereof. Furthermore, this specification provides a method for the polybioadhesive gel not to disrupt the ovarian function of the subject.Furthermore, this specification provides a method by which a polybioadhesive gel reduces the pain of a subject by at least 10% compared to the pain identified by the subject before the polybioadhesive gel was administered vaginally. Further, this specification provides a method by which vaginal administration of the composition provided herein reduces the systemic concentration of the activator or a salt thereof in a subject by at least 10 times compared to oral administration of the activator or a salt thereof to the subject. Further, this specification provides a method by which vaginal administration of the composition provided herein reduces the systemic concentration of the activator or a salt thereof in a subject by at least 15 times compared to oral administration of the activator or a salt thereof to the subject. Further, this specification provides a method by which vaginal administration of the composition provided herein reduces the systemic concentration of the activator or a salt thereof in a subject by at least 20 times compared to oral administration of the activator or a salt thereof to the subject. Further, this specification provides a method by which vaginal administration of the composition provided herein reduces the systemic concentration of the activator or a salt thereof in a subject by at least 25 times compared to oral administration of the activator or a salt thereof to the subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 30 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 35 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 40 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 45 times compared to oral administration of the activator or a salt thereof to a subject.Further provided herein are methods for administering a composition provided herein via vaginal flow, thereby reducing the systemic concentration of the activator or a salt thereof in a subject by at least 50 times compared to oral administration of the activator or a salt thereof to a subject. Further provided herein are methods for localizing the activator in the peritoneal cavity upon vaginal administration, by which the activator or a salt thereof diffuses into the ascites fluid of the subject, and further reaches the pelvic tissue via vaginal absorption. Further provided herein are methods for localizing the activator in the peritoneal cavity upon vaginal administration, by which the activator or a salt thereof diffuses into the ascites fluid of the subject, and further reaches the pelvic tissue via vaginal absorption.

[0142] Provided herein are methods for treating a disease or condition in a subject, comprising administering a cream or composition provided herein vaginally to the subject, thereby treating the disease or condition. Further provided herein are methods for determining, suspecting, or diagnosing, a subject having an endometrial lesion. Further provided herein are methods for determining, suspecting, or diagnosing, a subject having infertility. Further provided herein are methods for reducing the systemic concentration of an activator or salt thereof in a subject by at least 10 times by vaginal administration of a composition provided herein compared to oral administration of an activator or salt thereof to the subject. Further provided herein are methods for reducing the systemic concentration of an activator or salt thereof in a subject by at least 15 times by vaginal administration of a composition provided herein compared to oral administration of an activator or salt thereof to the subject. Further provided herein are methods for reducing the systemic concentration of an activator or salt thereof in a subject by at least 20 times by vaginal administration of a composition provided herein compared to oral administration of an activator or salt thereof to the subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 25 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 30 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 35 times compared to oral administration of the activator or a salt thereof to a subject. A method is further provided herein for administering a composition provided herein via vaginal artery to reduce the systemic concentration of the activator or a salt thereof in a subject by at least 40 times compared to oral administration of the activator or a salt thereof to a subject.This specification further provides a method for reducing the systemic concentration of the activator or a salt thereof in a subject by at least 45 times by vaginal administration of the composition provided herein compared to oral administration of the activator or a salt thereof to a subject. This specification further provides a method for reducing the systemic concentration of the activator or a salt thereof in a subject by at least 50 times by vaginal administration of the composition provided herein compared to oral administration of the activator or a salt thereof to a subject. This specification further provides a method for localizing the activator in the peritoneal cavity by allowing the activator or a salt thereof to diffuse into the ascites fluid of the subject upon vaginal administration, and to further reach the pelvic tissue via vaginal absorption. This specification further provides a method for localizing the activator in the peritoneal cavity by allowing the activator or a salt thereof to diffuse into the ascites fluid of the subject upon vaginal administration, and to further reach the pelvic tissue via vaginal absorption.

[0143] Provided herein are methods for maintaining ovarian function in a subject and reducing pain associated with endometriosis, comprising administering a cream or composition provided herein vaginally to the subject, thereby maintaining ovarian function in the subject and reducing pain associated with endometriosis. Further provided herein are methods for determining whether a subject has, is suspected of having, or is diagnosed with endometrial lesions. Further provided herein are methods for determining whether a subject has, is suspected of having, or is diagnosed with infertility. Further provided herein are methods for reducing the systemic concentration of the activator or a salt thereof in a subject by at least 10 times by vaginal administration of the activator or a salt thereof to the subject. Further provided herein are methods for reducing the systemic concentration of the activator or a salt thereof in a subject by at least 20 times by vaginal administration of the activator or a salt thereof to the subject. This specification further provides a method for reducing the systemic concentration of an activator or its salt in a subject by at least 25 times by vaginal administration of the composition provided herein compared to oral administration of the activator or its salt to the subject. This specification further provides a method for reducing the systemic concentration of an activator or its salt in a subject by at least 30 times by vaginal administration of the composition provided herein compared to oral administration of the activator or its salt to the subject. This specification further provides a method for reducing the systemic concentration of an activator or its salt in a subject by at least 35 times by vaginal administration of the composition provided herein compared to oral administration of the activator or its salt to the subject. This specification further provides a method for reducing the systemic concentration of an activator or its salt in a subject by at least 40 times by vaginal administration of the composition provided herein compared to oral administration of the activator or its salt to the subject. This specification further provides a method for reducing the systemic concentration of an activator or its salt in a subject by at least 45 times by vaginal administration of the composition provided herein compared to oral administration of the activator or its salt to the subject.This specification further provides a method for administering a composition provided herein via vaginal delivery, which reduces the systemic concentration of the activator or a salt thereof in a subject by at least 50 times compared to oral administration of the activator or a salt thereof to a subject. Furthermore, this specification provides a method for vaginal delivery in which the activator or a salt thereof diffuses into the subject's ascites fluid and further reaches the pelvic tissue via vaginal absorption, thereby localizing the activator in the peritoneal cavity. [Examples]

[0144] Examples Example 1: Formulation of a hormone-containing composition for transvaginal delivery. Exemplary pharmaceutical compositions containing bioadhesives for hormone delivery can be prepared for vaginal administration to subjects using the following formulations:

[0145] Progesterone preparations (%w / w): Micronized progesterone (USP / EP) 8.0% Carbomer (Carbopol 974P NF) 1.0% Polycarbophil Noveon AA1 (USP) 1.5% Sorbic acid (USP / EP) 0.1% Labrafac Lipophile WL 1349(USP / EP)17.07% EmulFree P Pharmaceutical Grade 2.0% Purified water (USP / EP) 68.4% Sodium hydroxide (USP / EP) QS AD pH 2.8-3.2 Purified water (USP / EP) QS 100%

[0146] A progesterone preparation can be prepared by dissolving micronized progesterone in an oil phase consisting of LABRAFAC® lipophille WL 1349 and EMULFREE® P. Alcohol may be added to this suspension at a concentration that does not function as a penetration enhancer. The carbomer is hydrated in the aqueous phase. After hydration, sorbic acid, polycarbophil, and sodium hydroxide are added to the aqueous phase. The two phases are mixed in a suitable container. The resulting composition is stored in a sealed container before vaginal administration.

[0147] In addition to progesterone P(4), any synthetic progestin can be formulated using the described formulation.

[0148] Estradiol preparations: (%w / w) Micronized estradiol (USP / EP) 0.1% Carbomer (Carbopol 974P NF) 1.0% Polycarbophil Noveon AA1 (USP) 1.5% Sorbic acid (USP / EP) 0.1% Labrafac Lipophile WL 1349(USP / EP)17.07% EmulFree P Pharmaceutical Grade 2.0% Purified water (USP / EP) 76.4% Sodium hydroxide (USP / EP) QS AD pH 2.8-3.2 Purified water (USP / EP) QS 100%

[0149] For example, estradiol can be substituted with any estrogen, such as dienestrol, estriol, or estrone.

[0150] Compositions containing bioadhesion substances and other active ingredients can be prepared in the same manner as the above-described formulations. Other active ingredients may include other hormones; antineoplastic agents; receptor agonists or antagonists; steroids; antibiotics; antiviral compounds; antifungal compounds; and anti-inflammatory agents. Example 2: Formulation of a vaginal administration composition containing danazol:

[0151] Exemplary pharmaceutical compositions containing bioadhesives for delivering danazol can be prepared for vaginal administration to subjects using the following formulations:

[0152] Danazol formulations: (%w / w) Danazol 6.67% Carbomer (Carbopol 974P NF) 1.0% Polycarbophil Noveon AA1 (USP) 1.5% Methylparaben 0.25% Labrafac Lipophile WL 1349(USP / EP)15% EmulFree P Pharmaceutical Grade 2.0% Purified water (USP / EP) 73.58%

[0153] Danazol formulations can be prepared by dissolving danazol in an oil phase consisting of LABRAFAC® lipophille WL 1349 and EMULFREE® P. Carbomer is hydrated in the aqueous phase. After hydration, methylparaben and polycarbophil are added to the aqueous phase. The two phases are mixed in a suitable container. The resulting composition is stored in a sealed container before vaginal administration.

[0154] Example 3: A comparative, open-label, randomized, parallel-group study to determine the efficacy of vaginal application formulations containing danazol as a treatment for endometriosis. To determine the intraperitoneal fluid, tissue, and serum concentrations of VML-0501 (100 mg of danatrol) after 5 days of daily vaginal application of a single dose, a comparative, open-label, randomized, parallel-group study was conducted in two groups of 12 women, each suspected or confirmed to have endometriosis and scheduled for laparoscopy, compared to 5 days of danatrol treatment administered as an oral capsule (danatrol) at a daily dose of 600 mg.

[0155] Study Objectives: The objective of this study was to evaluate the bioavailability of VML-0501 compared to oral danazol in the following respects: serum danazol concentration; and ascites fluid danazol concentration. Another objective of the study was to access the concentration of danazol in endometrial tissue (pathological tissue) found outside the uterus. A further objective of the study was to access the concentration of danazol in endometrial tissue. To access the concentration of danazol in myouterine tissue (only in patients where there was no increased surgical risk).

[0156] This was a comparative, open-label, randomized, parallel-group study investigating the concentrations of danazol when administered orally and vaginally. Subjects who met the inclusion and exclusion criteria were randomized in an open manner to receive either VML-0501 (vaginal danazol) or danatrol® (oral danazol) in a 1:1 ratio.

[0157] In the VML-0501 group, subjects were administered 1.5 g of VML-0501 cream (containing 100 mg of danazol) deep into the vagina using an internal applicator, followed by a 30-minute supine position. The subjects received the cream once daily in the morning for 5 to 7 consecutive days until the day of laparoscopy.

[0158] Subjects in the danatrol group received 200 mg of oral danatrol three times daily (TID) for 5-7 consecutive days leading up to the laparoscopy. Subjects participated in follow-up visits 7 / 8 days after discharge.

[0159] Number of subjects: The actual number of registered patients was 30.

[0160] Table 2 below includes the patient inclusion and exclusion criteria for the clinical trial. [Table 2]

[0161] Each subject received a 1.5-gram dose of VML-0501 cream once daily via vaginal administration, equivalent to 100 mg of danazol. The reference drug in this study was danatrol, containing 200 mg of danazol per capsule. Danatrol was administered orally three times daily.

[0162] Treatment period: All patients were to receive the assigned danazol treatment for 5 to 7 days or, based on the investigator's decision.

[0163] Assessment criteria:

[0164] Efficacy: The primary efficacy endpoint was measuring the concentration of danazol in ascites fluid and serum. The secondary endpoint was measuring the concentration of danazol in extrauterine endometrial tissue (lesional tissue).

[0165] Midway through the study, as part of a protocol modification, an exploratory endpoint was added, including the assessment of danazol concentrations in endometrial and myouterine tissue (only in patients without increased surgical risk).

[0166] We developed a bioanalytical method to test the concentration of danazol in human serum albumin and assessed the serum concentration (steady state) after administration, as well as the lesional concentration in tissue samples collected from ascites and laparoscopy.

[0167] Safety: Safety assessments included analysis of adverse events (AEs), physical examination, vital signs, blood, and urine tests to investigate the effect of the study drug on clinical tolerability. Adverse events were coded using WHO adverse reaction terminology. Adverse events were reported by major physical system and preferred terminology.

[0168] Statistical methods:

[0169] Descriptive statistics were used to summarize all continuous variables (such as clinical demographic features), including patient count, mean, standard deviation, median, first quartile, and third quartile. For categorical variables by category, absolute numbers and percentages were presented. Primary and secondary endpoints were summarized in tabular summaries and lists. Means and 95% confidence intervals (CI) or median and interquartile range (depending on the nature of the parameter) were investigated for each endpoint. Paired Student's T-tests and nonparametric paired Wilcoxon tests were used to compare serum and ascites within each group. Wilcoxon's Mann-Whitney U test was used to compare the ratio between ascites and serum between two groups. Due to surgical risks, tissue samples could not be collected from all patients with detectable danazol in both serum and ascites. Due to the small number of tissue samples collected, data were qualitatively described to report the number of patients with detectable danazol in tissue samples within each treatment group.

[0170] Adverse events and concomitant medications were reported for each patient. The denominator in percentage calculations represented the number of patients in the target population. Subject selection for inclusion in the analysis was defined as follows: Primary population - Treatment Intent - (Subjects selected, randomized, and who received at least one dose of the study drug); Secondary population - Protocol Compliant - (Subjects who completed the study trial and received at least 80% of the study drug); Package SAS® System version 9.4 was used for all analyses. A p-value of <0.05 was considered statistically significant.

[0171] Example 4: Clinical efficacy of the danazol composition. Of the 30 patients enrolled in this study, 29 were randomized (16 to the VML-0501 group and 13 to the danatrol group). Subsequently, two patients withdrew their consent and two patients dropped out for other reasons. Baseline demographics and characteristics were homogeneous between the VML-0501 and danatrol groups. Comparison of serum concentrations between the two groups showed that serum concentrations were significantly lower in the vaginal group (p<0.001). Comparison of danatrol concentrations within each group showed that concentrations were higher in serum than in ascites (p<0.001), both when values ​​<1 were considered as 0.5 or missing.

[0172] Similar ratios of danazol concentration from serum to ascites fluid were reported in both groups, confirming similar systemic effects, which were attributed to different mechanisms of action when VML-00501 was administered vaginally. Danazol concentrations were significantly higher in tissue than in serum samples in the VML-0501 group (n=6, p-0.031), but not significantly higher in the danatrol group (n=11, p=0.054).

[0173] Example 5: Safety of the danazol composition. No serious adverse events (SAEs) or photoallergic reactions were observed in any of the treatment groups. Only three AEs were recorded among all registered subjects (two in the VML-0501 group and one in the danatrol group). There were no significant differences in AEs between the groups.

[0174] VML-0501, a polybioadhesive gel, provides sustained benefits consistent with its direct tissue effects on lower pelvic inflammation. Low serum levels do not affect the hypothalamic-pituitary axis, protecting ovarian function, minimizing systemic effects, and improving fertility.

[0175] Results from the VML-0501-001 study show significantly lower serum danazol concentrations in VML-0501 compared to oral danazol. The use of low-dose vaginal danazol reaching ascites and the peritoneal cavity should have beneficial effects on ovarian quality, reducing inflammation and immunomodulating pain signaling in ascites. Danazol concentrations in tissue biopsies were significantly higher in the VML-0501 group compared to serum levels, suggesting drug diffusion into ascites, further reaching pelvic tissue via transvaginal absorption and countercurrent migration between uterine lymphatic vessels or veins and arteries, and retaining the drug in that area.

[0176] Example 6: A method for treating endometriosis. Endometriosis is a condition in women characterized by the presence of endometrial tissue (endometrial glands and stroma) outside the uterine cavity, and is commonly associated with chronic pelvic pain and infertility. Examples of these lesions may be peritoneal lesions or deep invasive disease. Diagnosis is often delayed due to the lack of consistent diagnostic biomarkers for endometriosis. The effectiveness of treatments such as hormone therapy and analgesics used for symptomatic endometriosis is limited due to the high recurrence rate in many women.

[0177] Oral danazol is a treatment for endometriosis. Danazol is a synthetic steroid, a derivative of 17-alpha-ethinyltestosterone, and is used to suppress ovarian function by forcing a direct inhibitory effect on ovarian steroid production and by inhibiting gonadotropins (follicle-stimulating hormone (FSH) and luteinizing hormone (LH)). Danazol has mild androgenic effects but lacks estrogenic or progesteroneic properties. In women with regular menstrual cycles, danazol induces a state of pseudomenopause characterized by complete suppression of ovarian function, amenorrhea, and a hypoestrogenic state. Endometrium and ectopic endometrium undergo atrophy during treatment, resulting in a reduction of endometriosis.

[0178] Provided herein is a method for treating endometriosis, comprising vaginal administration of 100 mg of danazol in the form of a liquid emulsion. Provided herein is a first comparative study analyzing different concentrations of 100 mg of vaginal-administered danazol (VML-0501) in serum, ascites, and endometrial tissue compared to 600 mg of oral danazol capsules (danatrol®).

[0179] Objective: The primary objective of this study was to investigate and compare the concentrations of danazol in the serum and ascites fluid of patients in both the VML-0501 group and the danatrol group. A secondary objective of this study was to investigate the concentrations of danazol in lesional tissues. The objective of this study was to investigate the concentrations of danazol in the endometrium and uterine muscle tissue (where applicable).

[0180] Research plan Overall research design and plan The VML-0501-001 study was a phase II parallel-arm, open-label, single-center proof-of-concept study designed to determine the levels of danazol concentration in patients suspected of or clinically diagnosed with endometriosis, in response to oral or vaginal danazol treatment. The planned enrollment was approximately 24 patients, and was later expanded to 34 patients due to difficulties in recruiting the patient population.

[0181] Patients suspected of having or clinically diagnosed with endometriosis were screened and encouraged to participate in the study during routine hospital visits. Informed consent was obtained on day 1, and patients proceeded for a physical examination. Patient height and weight were measured, and vital signs were recorded. Blood and urine samples were collected for baseline analysis, and pregnancy tests were completed to confirm eligibility. All trials were conducted as part of standard care during patients' routine hospital appointments. Patients were informed to participate on day 4 of their menstrual cycle, where they were randomized to either the VML-0501 group or the danatrol group.

[0182] A physical examination was completed, and vital signs were recorded during this visit. Patients were provided with instructions on how to complete each dose over the following 5-7 days prior to the laparoscopy day (for some patients, the procedure was extended to 7 days at the medical discretion of the investigator). Laparoscopy and sampling were performed prior to day 14 of the menstrual cycle (the day of laparoscopy varied among patients due to the severity of bleeding during the menstrual cycle, which determined the initiation date and procedural selectivity of the investigational procedure).

[0183] Steady-state sampling is achieved within 5 days after administration, and was therefore completed on either day 8, 9, or 10 of the menstrual cycle.

[0184] The patient was discharged after the laparoscopy and asked to return for a follow-up visit approximately one week after discharge. During the follow-up visit, the patient provided blood and urine samples and underwent a physical examination.

[0185] Safety was assessed based on patient reports in patient workbooks at every study visit and throughout the study. Adverse events were closely monitored, and their incidence, severity, and relationship to the study drug were evaluated. Figure 2 provides a schematic diagram of the study design, showing the timing of study evaluation and administration.

[0186] Study design including selection of control group The treatment was administered in a parallel, open-label manner. Both patients and investigators were open-label regarding the treatment assignment. This design was chosen to minimize the risk of treating patients with ineffective treatments, and early termination / withdrawal from the study was permitted if desired by the patient or investigator. A total of 34 patients were expected to be randomized in each arm, with 17 patients per arm.

[0187] The control group selected for this study was the danatrol arm. Danatrol, an approved drug, is a derivative consisting of 200 mg of danazol as the active substance per capsule. Therapeutic indications for danatrol include the symptomatic treatment of endometriosis as an oral administration. This form of treatment should be initiated during menstruation in women who are not pregnant or lactating, in conjunction with an effective nonhormonal contraceptive. To achieve amenorrhea, an initial dose of danatrol of 600 mg / day should be administered starting from the first day of the menstrual cycle, followed by a maintenance dose as determined by a healthcare professional. This form of danazol is absorbed from the gastrointestinal tract and then reaches a peak plasma concentration of 50–80 ng / ml approximately 2–3 hours later [4]. Studies have shown that bioavailability can be tripled by a high-fat diet, but absolute bioavailability remains unknown. Danazol is metabolized very rapidly and almost completely in the liver. Vaginal application of danazol may bypass the liver to reduce any undesirable effects. This selection of the control group was based on evidence that low-dose danazol treatment has shown positive results in alleviating endometriosis symptoms, which was used as a comparison in this study for the same, if not improved, desirable outcomes.

[0188] To ensure an equal comparison of danazol concentrations between oral and vaginal doses, this study used a parallel design.

[0189] research group Inclusion Criteria

[0190] To be eligible for inclusion in the study, subjects had to meet the following criteria:

[0191] Women between the ages of 18 and 42.

[0192] Prior to the completion of any research-related procedures, you may provide written (signed and dated) informed consent (this means any evaluation or assessment that does not normally form part of medical treatment).

[0193] You have or are suspected of having endometriosis.

[0194] I am scheduled to undergo a laparoscopy.

[0195] According to local practice, women should sign a specific clause regarding pregnancy prevention based on the use of two effective methods of contraception (condoms and spermicides) 30 days after the last dose / application.

[0196] She is a non-pregnant woman who has undergone laparoscopy for confirmed or suspected endometriosis within the first 14 days of her cycle.

[0197] Body Mass Index (BMI) ≤ 32 kg / m2

[0198] Exclusion criteria:

[0199] If any of the following criteria apply, the subject could not be enrolled in the study:

[0200] Pregnant (positive urine pregnancy test during screening) or breastfeeding.

[0201] They have evidence of drug or alcohol abuse.

[0202] You have used hormone replacement therapy or danazol therapy within the past 8 weeks prior to participating in the study.

[0203] Having any of the following: epileptic seizures, migraines, angina pectoris, chronic heart failure, vascular occlusion due to blood clots, liver problems, kidney disease, pregnancy, a combination of high blood cholesterol and triglyceride levels, or porphyria.

[0204] Undiagnosed abnormal genital bleeding.

[0205] Androgen-dependent tumors.

[0206] They are allergic to anabolic and masculinizing steroids.

[0207] Removal of subjects from treatment or research

[0208] In accordance with the research protocol, withdrawal or early termination was defined as withdrawal from the research procedure after initial exposure to the research treatment.

[0209] Subjects who dropped out before the first dose of either VML-0501 or danatrol were considered "screening failures." Subjects could withdraw from the study at any time at their own request or at the investigator's clinical discretion. No disadvantage was incurred by those who wished to withdraw their consent or who were excluded from the study by the investigator for any reason. Subjects who requested to withdraw from the procedure were strongly encouraged to complete appropriate tests for safety and tolerability assessment.

[0210] Participants who withdrew from the study for medical reasons or at the sponsor's request were not followed up on regarding the study (patients participated in SoC routines for hospital follow-up).

[0211] The following categories were used to classify the reasons for withdrawal:

[0212] Poor tolerability: Unacceptable poor tolerability reported by the subject or observed by the physician; a corresponding AE with a causal relationship classified as “possible” should be linked.

[0213] Intervention-related illness: Any illness or condition that occurred during the clinical trial and prevented further participation; however, there is no evidence of a causal relationship between the event and the research treatment; there should be corresponding AEs with a causal relationship classified as “unlikely” or “unassessable.”

[0214] Withdrawal of consent / Non-compliance: It is checked whether the subject withdrew consent or failed to comply with the research procedure; whether an adverse event may have caused withdrawal.

[0215] Other reasons: Reasons for discontinuation not covered by any of the above criteria, such as the subject moving, the indication no longer being applicable, or other reasons must be identified.

[0216] Treatment administered: VML-0501 or danatrol was administered to patients who met the entry criteria at either 100 mg of danatrol once daily or 200 mg of danatrol three times daily, for 5 to 7 consecutive days over one study period. The length of treatment varied among patients based on the length of their menstrual cycle and medical determination of their PI. The treatment administered was based on which arm of the study the patient was randomized to. Treatment continued for each patient until the day of laparoscopy.

[0217] VML-0501: Danazol is a synthetic steroid (chemical compound) with antigonadotropin and antiestrogenic activity that acts as an anterior pituitary inhibitor by inhibiting the pituitary clearing of gonadotropins. It possesses several androgenic properties. Danazol is used to treat endometriosis and several benign breast disorders. Danazol is a class II molecule (low solubility, high permeability) according to the G. Amidon classification (BCS). Log P = 4.5 (testosterone = 3.22, progesterone = 3.82, estradiol = 4.01). It was approved by the U.S. Food and Drug Administration in 1971 as the first drug to specifically treat endometriosis and is widely marketed worldwide, including in Italy where this research was conducted.

[0218] Danazol exists as a white to pale yellow crystalline powder. VML-0501 is a cream developed for vaginal application and contains 6.7% (w / w) danazol as the active ingredient. It is formulated with polycarbophil, sorbic acid (potassium sorbate), purified water, Labrafac (propylene glycol dicaprylate), Carbomer 974P, and Emulfree (propylene glycol laurate (and) ethylcellulose (and) propylene glycol isostearate). It is supplied as a 30g aluminum tube with a vaginal applicator. Quantitative formulations of VML-0501 are provided in Table 3 below. [Table 3]

[0219] The storage conditions for VML-0501 were below 25°C and 65±5% relative humidity, in accordance with ICH guidelines. GB Pharma confirmed that the IMP was stored in a dry, cool place away from direct sunlight.

[0220] The decision to use 100 mg of danazol in VML-0501 was made because this study requires a low dose to investigate the effects of oral administration on serum, ascites, and tissue. Comparison: Danatrol

[0221] Danatrol comparator was obtained from a local pharmacy (as this product is commercially available) and was never altered. The supplies were sent to MiPharm, which labeled them according to the clinical trial protocol and delivered the drugs to the hospital pharmacy. The 100 and 200 mg danatrol capsules contain corn starch, lactose, talcum, and magnesium stearate as excipients. The capsule shell contains titanium dioxide (E171) and gelatin. Danatrol has a 5-year stability and was not stored at temperatures above 25°C in accordance with the manufacturer's guidelines. The known plasma half-life of danatrol varies between 3 and 6 hours after a single dose.

[0222] Dosage selection and timing A daily dose of 100-200 mg of oral danazol may result in symptom relief, reduction of severe menstrual bleeding and lesion volume, and potential improvement in fertility in patients with endometriosis. In more severely affected patients, higher doses of oral danazol (600-800 mg) may be required daily, but associated side effects have led to the limited use of this treatment.

[0223] In this specification, danazol administered vaginally was tested to determine its usefulness as a drug for the treatment of rectovaginal endometriosis.

[0224] Vaginal application allows for direct targeting of lesions near the vagina at lower doses than those required for oral administration. The danazol formulations provided herein result in higher concentrations of the drug distributed in the surrounding area and lower levels of circulating drug. Vaginal application also minimizes the side effects frequently associated with oral administration and does not inhibit ovulation. Vaginal administration also limits systemic transmission of danazol and thus minimizes androgenic side effects. This procedure has been shown to be well-tolerated, with patient satisfaction reaching 80%, and the average consumption of pain-relieving tablets (e.g., naproxen) was reduced by 60% in these previously refractory patients. However, its use, even at lower doses, is contraindicated during pregnancy. Vaginal danazol can be offered as an adjuvant to conventional medical procedures, avoiding the contraceptive issues that may arise with these procedures. It can also be an alternative to performing difficult surgery in patients with deep-invasive endometriosis of the rectovaginal septum.

[0225] For a selected population of clinically diagnosed patients suspected of having endometriosis, the appropriate daily dose of vaginal danazol in IMP was determined to be 100 mg (1 / 6 of the total control dose), and danatrol was determined to be 600 mg for appropriate comparison. The study dosing regimens are summarized in Table 4 below. [Table 4]

[0226] Blind This was an unblinded, parallel-arm, open-label proof-of-concept study. The objective of this study was to obtain further information on the safety and mechanism of action of VML-0501 using a novel delivery method different from conventional oral therapy.

[0227] Previous medications and concomitant medications All concomitant medications were continuously reported from the start of treatment to the final follow-up visit, as required by the protocol.

[0228] The use of topical or systemic hormone replacement therapy (HRT) containing estrogen within the weeks preceding the procedure was not permitted. The use of topical and systemic estrogen phototherapy was also not permitted. Participants were required not to use lubricants during sexual intercourse during the study period.

[0229] Patients were able to continue medication for any underlying conditions that were already present at baseline (Visit 1) and did not disqualify them from the study. Prior treatment was reported only if it was significant and contemporaneous at enrollment.

[0230] Compliance with procedures Participants received treatment at the study facility and self-administered the drug at home until their scheduled laparoscopy appointment. Study procedure compliance was assessed through dose administration, accident reporting, and drug accountability monitoring upon receipt of assigned IMP / comparison (empty / partially / unused filled tubes / boxes) from participants at the end of the study.

[0231] The monitors verified that the receipt and batch numbers of all drug supplies at the site matched the corresponding drug accountability records.

[0232] A complete list of the research procedures is provided in Table 5 below. [Table 5] 1. Apply VML-0501 daily at 8:00 AM until the laparoscopy. 200 mg of oral danazol (danatrol) is applied three times daily until the laparoscopy. 2. Laparoscopy and sampling can be performed on any of the 8th, 9th, or 10th days of the menstrual cycle. 3. Steady-state sampling is achieved on day 5 of administration, so it can be performed on day 8, 9, or 10 of the menstrual cycle. 4. Four follow-up visits can be scheduled 7 / 8 days after discharge.

[0233] Several assessments were performed during the screening period to determine patient eligibility. Local pathological reviews and diagnoses were used to determine whether patients met the requirement of being suspected or clinically diagnosed with endometriosis according to the inclusion criteria. Trained healthcare professionals and investigators were responsible for screening patients to determine their eligibility.

[0234] Furthermore, urine pregnancy tests were performed on women of childbearing age within three days prior to the first dose.

[0235] Medical history and physical examination Medical history assessment and physical examination were completed as part of standard care within 4 days prior to the planned initiation of danazol administration, and the following data were recorded: date of birth, age, height, weight, BMI, blood pressure, heart rate, any ongoing medical conditions, and details of medications taken prior to danazol administration (the relevance of ongoing conditions and medications was assessed by the investigator for danazol suitability).

[0236] Effectiveness measurement Efficacy was assessed by examining the concentration levels of danazol in serum, ascites, and tissue. Hematological and serological evaluations, coagulation, and urinalysis were performed at baseline (within 3 days prior to the planned start of danazol administration) and during the final follow-up visit. Thus, comparisons could be made both before and after danazol administration.

[0237] The primary efficacy endpoint analysis was predefined as a comparison of danazol concentrations in the aforementioned parameters within and between patients in both arms of the study (VML-0501 vs. danatrol).

[0238] Adverse events Symptoms, adverse events (AEs), and co-occurring illnesses were monitored throughout the study and recorded at each visit. AEs were coded according to the Medical Dictionary for Regulatory Activities (MedDRA®) coding system, based on the event and disease history.

[0239] Laboratory measurements

[0240] Laboratory assessments were performed as part of standard care during baseline and end-of-study follow-up visits. Based on the patient's menstrual cycle and investigator's decision, blood samples were collected on either day 8, 9, or 10 of the study for drug steady-state analysis. Oral danazol is known to reach steady state within 7 days of administration.

[0241] Vital signs

[0242] Blood pressure and heart rate were measured for all patients at baseline, on day 1 of administration, and during discharge as part of standard care.

[0243] Effectiveness variable

[0244] For the biological analysis and quantification of danazol in human serum albumin, assays validated according to EMA and FDA standards were used to assess the primary efficacy endpoint for determining danazol concentrations. Measurements were compared between serum and ascites, as well as within the groups, as individual comparisons between the VML-0501 group and the danatrol group. Secondary efficacy endpoints included measuring danazol concentrations in endometrial tissue (lesional tissue) observed outside the uterus, and, if there was no increased surgical risk in patients, investigating danazol concentrations in endometrial and myofascial tissue as exploratory endpoints.

[0245] Safety variables

[0246] Safety variables included the incidence and severity of adverse events and serious adverse events (SAEs), concomitant medications, early withdrawal, physical examination parameters, vital signs, menstrual cycle, and routine clinical laboratory tests (blood and urine tests by SoC).

[0247] Statistical and analytical planning VML-0501 was a Phase II parallel-arm open-label study. The primary objective and endpoint of the study was to determine the concentrations of danazol in the serum and ascites fluid of patients suspected of or clinically diagnosed with endometriosis. The dataset was analyzed and reviewed for a total of 30 patients.

[0248] The clinical demographic characteristics of the patients were summarized in lists and tables as mean, standard deviation (sd), median, and interquartile range (Q1-Q3) for all continuous variables. For categorical variables, the data were listed as absolute number (n) and percentage. These were compared separately for the VML-0501 group and the danatrol group.

[0249] The mean and 95% confidence interval or median and interquartile range (depending on the nature of the parameters) were calculated for each primary and secondary endpoint. The primary endpoint was compared between the VML-0501 group and the danatrol group using the T-Student test and the Mann-Whitney U test.

[0250] Safety analyses were reported by a list of adverse events and concomitant medications reported for each subject. SAS® System package, version 9.4, was used for all analyses. A p-value of <0.05 was considered statistically significant.

[0251] Determining the sample size As this was a proof-of-concept study, the sample size of 34 patients was selected based on practical considerations rather than statistical considerations. VML-0501 is in the early stages of clinical drug development. This was a POC study to demonstrate that vaginal delivery of 100 mg of danazol, which is 1 / 6 of the oral dose, can potentially reach similar therapeutic pelvic concentrations (pelvic tissue and ascites) as 600 mg of oral administration (oral danazol 200 mg capsule TID).

[0252] The subjects selected for the analysis are as follows:

[0253] Safety Aid Set (SAF): This included all subjects who were enrolled in the study and administered at least one dose of the investigational drug.

[0254] Primary population: Intentional Treatment (ITT). These are all subjects selected, randomized, and administered at least one dose of the study drug.

[0255] Secondary population: Protocol-compliant (PP). These are all subjects who completed the study and ingested at least 80% of the study drug.

[0256] Effectiveness assessment Analyzed dataset The safety set (N = 30) included all subjects who were registered in the study and received at least one dose of the investigational drug.

[0257] The intention-to-treat (ITT) (N = 29) population included all subjects who were selected, randomized, and received at least one dose of the study drug.

[0258] The per-protocol (PP) (N = 26) population included all subjects who completed the study and took at least 80% of the study drug.

[0259] Demographics Table 6 below summarizes the patient demographics of the ITT population. All patients were women between 25 and 47 years old. The median age was 35.0 for the entire ITT population, and the mean BMI was 21.8.

Table 6

[0264] When comparing danazol concentrations in serum samples from both the VML-0501 group and the danatrol group, serum concentrations were significantly lower in the VML-0501 group (P<0.001). However, within each treatment group, serum concentrations were higher than those in ascites fluid (p>0.001), and any value less than 1 in both cases was considered 0.5 or missing.

[0265] The ratio of danazol concentrations in serum and ascites fluid did not differ significantly between the VML-0501 group and the danatrol group (p=0.417 when <1 is set to 0.5; p=0.369 when <1 is set to missing). [Table 8]

[0266] A comparison of serum concentrations between the VML-0501 group and the danatrol group concluded that the mean serum concentration was significantly lower in the VML-0501 group, approximately 40 times on average (p<0.001). The ratio of danazol concentration between serum and ascites fluid did not differ significantly between the VML-0501 group and the danatrol group (p=9.640 when <1 is set to 0.5, p=0.350 when <1 is set to missing). [Table 9] [Table 10]

[0267] The secondary endpoint was to investigate the concentration of danazol measured in endometrial tissue. Due to the surgical risk, tissue samples could not be collected from all 26 patients with detectable levels of danazol. Insufficient sample mass was collected from 4 out of 15 patients (27%) in the VML-0501 group.

[0268] Detectable values of danazol were available for 6 out of 11 patients (55%) (range 1.3 - 5) in the VML-0501 group and 11 out of 11 (100%) (range 3.2 - 541) in the danatrol group. The concentration of danazol was significantly higher in tissue than in serum samples in the VML-0501 group (n = 6, p = 0.031), but did not reach significance in the danatrol group (n = 11, p = 0.054).

[0269] In summary, it is clear that when administered vaginally using VML-0501 cream, the level of danazol in serum is lower compared to orally administered danazol.

[0270] Figure 3 shows a graph of the ratio of danazol concentration between serum and ascites in the (ITT) population.

[0271] Figure 4 shows a graph of the ratio of danazol concentration between serum and ascites in the PP population.

[0272] Figure 5 shows a graph of the concentration of danazol in serum and tissue in the VML-0501 treatment group.

[0273] Figure 6 shows a graph of the concentration of danazol in serum and tissue in the oral danazol group.

[0274] Example 8: Safety assessment of VML-0501. All safety analyses were performed on a full analysis dataset (safety set) that included all patients enrolled in the study and included all subjects who received at least one dose of VML-0501.

[0275] The following table provides an overall overview of the degree of danazol exposure for the safety set. [Table 11]

[0276] Brief summary of adverse events: Adverse events were collected for all patients, regardless of causal relationship, from the time of initial administration to the date of the final follow-up visit. A general overview of the AEs observed in this study is provided in Table 13. Adverse events are defined as any unfavorable medical occurrence in patients administered the investigational drug, which is not necessarily causally related to the treatment itself.

[0277] For 95% of subjects using the Clopper-Pearson Cis, the proportion of subjects experiencing at least one adverse event was calculated for all AEs and differentiated by treatment group. Of the 30 subjects enrolled, a total of 3 experienced adverse events in this study.

[0278] Each of the three subjects reported one adverse event (AE) (10.3%, CI 95%, 0.02-0.27), two events were reported from the VML-0501 group (12.5%, CI 95%, 0.02-0.38), and one event was reported from the danatrol group (7.69%, CI 95%, 0.00-0.36). [Table 12]

[0279] No serious adverse events were reported. In conclusion, there were no safety concerns in either treatment group. There were no statistically significant differences in the incidence of adverse events between the two groups. No deaths or SAEs were reported as a result of either treatment.

[0280] Example 9: Comparison of vaginal and oral delivery of danazol. Oral danazol reduces ovarian estrogen production and endometrial lesions, alleviating the symptoms of endometriosis, but causing unpleasant androgenic side effects. Danazol has high binding affinity to progesterone and androgen receptors, which have a direct effect on endometrial lesions, leading to induced apoptosis (programmed cell death). This can occur as a result of disruption of secondary messaging and DNA synthesis at the cellular level. Generally, aside from the pain symptoms associated with endometriosis, up to 50% of women experiencing infertility have symptoms related to endometriosis. In fact, biochemically, endometriosis can have detrimental effects on oocyte quality or endometrial receptivity.

[0281] Current treatments for endometriosis focus on systemic ovarian suppression, which depletes the growth capacity of endometriotic lesions. This treatment induces a transient chemical menopausal state in women, resulting in a lack of ovulation, bone loss, mood swings, facial flushing, and other potential debilitating effects. This leads to the need for "additional" treatment to counteract these unintended effects. Further molecular effects of danazol include, for example, improved activity of immune cells (e.g., natural killer cells), increased Fas-induced apoptosis of endometrial cells, and anti-inflammatory effects in the pelvic cavity. Oral danazol is metabolized in the liver by enzymes such as CYP3A4 and can also elevate other liver enzymes. Patients receiving danazol capsules at a daily dose of 400 mg or more may have elevated serum enzymes and / or jaundice.

[0282] As provided herein, bypassing the liver via targeted vaginal administration of danazol can reduce the undesirable side effects of hepatic impairment and hepatotoxicity observed with oral danazol. Lower doses of vaginal-administered danazol have a better side effect profile and avoid most of the aforementioned adverse effects by bypassing the initial hepatic pathway.

[0283] VML-0501 utilizes targeted vaginal delivery technology to facilitate the uptake of danazol from the cervical application site into the peritoneal cavity. VML-0501 can target the pelvic organs with the drug while restricting systemic circulation. While not bound by any specific theory, this effect may be due to a countercurrent mechanism in which arteries and veins located in close proximity to each other flow in opposite directions, thereby preventing significant amounts of the drug from being absorbed systemically. Furthermore, VML-0501 is absorbed into the peritoneal cavity via transvaginal epithelial absorption. Danazol reaches the vaginal arteries, with minimal leakage into systemic circulation, and is drawn back into the pelvic cavity, peritoneal organs, and ascites. It is also transported from the capillary network (reticular reticularis) with countercurrent movement between uterovaginal lymphatic vessels or veins and arteries, aimed at maintaining locally high concentrations, enabling danazol binding to endometrial and ovarian tissue. This transcends the initial uterine transit metabolism. Therefore, the compositions and methods provided herein generate novel pharmacokinetic profiles of drugs, which are referred to herein as pelvic transit metabolism.

[0284] VML-0501 induces atrophy of endometrial lesions without the need to inhibit the ovarian-pituitary axis. This is achieved without systemic levels of danazol drugs on which other treatments rely, thus preventing debilitating side effects. In women with normal cycles, oral administration of danazol induces a state of pseudomenopause characterized by complete suppression of ovarian function, amenorrhea, and a hypoestrogenous state. Endometrium and ectopic endometrium undergo atrophy during treatment, resulting in regression and disappearance of endometriosis.

[0285] Results from the VML-0501-001 study showed significantly lower serum danazol concentrations compared to oral danazol. Serum concentrations in patients administered vaginally (16 / 16) were significantly lower than those in the oral administration group (Mann-Whitney p=0.0001233). The drug was detected in ascites fluid and quantified in most samples (12 / 16). Analysis of the ratio between measured serum concentrations and measured ascites concentrations showed no statistically significant difference between women administered VML-0501 and oral danatrol (Mann-Whitney p=0.2773).

[0286] The use of low-dose vaginal danazol that reaches ascites and the peritoneal cavity can have beneficial effects on ovarian quality, reduce inflammation, and immunomodulate pain signaling in ascites.

[0287] When pelvic biopsies (extrauterine tissue including ovaries, endometrial cysts, ovarian fossa, broad ligament, uterosacral ligament, and peritoneum) were obtained (12 / 16), danazol was detected in all samples. Danazol concentrations in tissue biopsies were significantly higher in the vaginal danazol group compared to serum levels (p=0.031). The same was not observed when danazol concentrations in pelvic tissue were analyzed in the oral group.

[0288] Similar ratios of danazol concentration from serum to ascites fluid were reported in both groups, confirming similar systemic effects and a mechanism quite different from that observed with vaginal administration of VML-00501. The limited leakage of danazol after vaginal administration of VML-0501 further protects patients from undesirable systemic side effects.

[0289] The relatively high concentrations achieved in pelvic tissue (via biopsy) suggest the diffusion of the drug into ascites fluid, further reaching pelvic tissue via transvaginal absorption, and countercurrent movement between the uterine and vaginal lymphatic vessels or veins and arteries retains the drug in that area.

[0290] Vaginal administration of danazol resulted in zero treatment-related adverse events or side effects. No serious adverse events were recorded in any arm of the study. This indicates that the vaginal administration route using VML-0501 cream can target endometrial lesions and provide symptom relief without the undesirable side effects seen with oral danazol administration.

[0291] The low serum concentrations of danazol administered vaginally were quite surprising for two reasons. Firstly, the pelvic cavity is highly angiogenic. Therefore, vaginal administration of VML-0501 was expected to produce serum concentrations of danazol comparable to those of oral administration of danatrol®. It was also expected that VML-0501 would produce serum concentrations of danazol similar to those of orally administered danatrol®, because vaginal VML-0501 administration was expected to bypass first-pass metabolism in the liver.

[0292] Surprisingly, VML-0501 had significantly lower serum danazol concentrations compared to orally administered danatrol®. Significantly lower serum concentrations result in fewer adverse effects.

[0293] Orally administered danatrol can cause a number of undesirable side effects, including cardiotoxicity and hepatotoxicity. Specifically, orally administered danatrol® is associated with undesirable effects such as acne, hair loss, fluid retention, weight gain, increased insulin resistance in diabetic patients, neurological disorders (e.g., anxiety and depression), nausea, and convulsions.

[0294] In contrast to oral administration, vaginal administration of VML-0501 resulted in zero treatment-related adverse events or side effects. No serious adverse events were recorded in any arm of the study. These results indicate that the vaginal administration route using VML-0501 emulsion targets endometrial lesions and provides symptom relief without the undesirable side effects seen with oral danazol administration. Thus, even unexpected results are considered excellent.

[0295] To determine the distribution of danazol in pelvic tissue between groups, pelvic biopsies (extrauterine tissues including ovaries, endometrial tumor cysts, ovarian fossa, broad ligament, uterosacral ligament, and peritoneum) were assessed from subjects in each group. Danazol was detected in ascites fluid and quantified in most samples (12 / 16) in both groups. Impressively, danazol concentrations in tissue biopsies were significantly higher in the VML-0501 group compared to the danatrol® group (n=6, p=0.031). The tissue distributions shown in Figures 4 and 7 also showed superior results, as VML-0501 consistently delivered higher sustained concentrations of danazol in the target tissue, while danatrol® delivered inconsistent and significantly lower concentrations in the target tissue. Prior to collecting this data, it was anticipated that vaginal application of VML-0501 would result in faster diffusion from the target tissue and lower sustained concentrations in the target tissue.

[0296] Further investigation of the distribution of danazol between groups revealed that clinical trial data showed that vaginal-administered VML-0501 maintained high and consistent concentrations of danazol within the peritoneal tissue via diffusion into ascites fluid, reaching the pelvic tissue via vaginal absorption and countercurrent movement between the uterine lymphatic vessels, veins, and arteries. Therefore, the mechanism by which our vaginal-administered composition reached the target pelvic tissue was significantly different from that of orally administered danazol. Next, vaginal administration of danazol kept it out of systemic circulation within the targeted pelvic region (Figure 7). Consequently, vaginal-administered VML-0501 resulted in unexpectedly and significantly higher sustained target tissue danazol concentrations.

[0297] The results described in the above examples demonstrate that a method of administering a pharmaceutical composition in the form of a danazol emulsion to a subject via vaginal delivery can reduce the side effects associated with orally administered danazol. The above results also demonstrate that the method provided herein also reduces the side effects typically associated with oral administration of danazol. Finally, the results show that vaginal administration of VML-0501 also provides a high and sustained concentration of danazol in the target tissue.

[0298] Example 10: A method for treating pain and infertility using VML-0501. Medical intervention is not recommended when the primary objective is to improve fertility in patients with endometriosis. This is because all current medications used to treat endometriosis are hormonal and therefore inhibit ovulation. In contrast, medical intervention is beneficial when the primary objective is to target pain. Endometriosis is considered a long-term disease, its natural history is unknown, and long-term management may be necessary depending on the patient's age, symptom profile, and desire for fertility.

[0299] Infertility associated with endometriosis may be linked to mechanical pelvic anatomical distortions that potentially impair the "uptake" of oocytes by the fallopian tubes. Biochemically, endometriosis can have detrimental effects on oocyte quality or endometrial receptivity. Treatment options for women trying to conceive are either waitlist management, surgery, or assisted reproductive technology (ART).

[0300] This specification provides a method for treating infertility and pelvic pain using VML-0501. The use of low-dose vaginal danazol is highly effective in treating pain symptoms because it aims to reduce inflammation and immunomodulate pain signaling in ascites fluid.

[0301] VML-0501 is also used to directly reduce the size of endometrial lesions by increasing apoptosis of extrauterine endometrial cells (cytopathic effect) and by starving the lesions, theoretically without systemic side effects, by locally reducing aromatase P450 expression.

[0302] Danazol modulates the function of a normal human endometrial stromal cell subpopulation by altering the endometrial cytokine network. This results in a "reprogramming" effect that all begins at the orthotopic endometrial level.

[0303] The polybioadhesive gel VML-0501 provided herein offers sustained benefits consistent with a direct tissue effect against lower pelvic inflammation. Low serum levels do not affect the hypothalamic-pituitary axis, protect ovarian function, minimize systemic effects, and improve fertility.

[0304] Assays can be performed to determine the optimal dose of VML-0501 while assessing the safety and efficacy of dose comparisons. The molecular mechanism of action behind the fertility aspect of danazol can be investigated in more detail, focusing on telomerase activity. Telomerase is active during embryogenesis and is released by endometrial cells in the later stages of the menstrual cycle in women with endometriosis, resulting in abnormally long telomere lengths in these women. While not bound by any specific theory, vaginal danazol administration is expected to have a direct effect at various levels (ovarian and orthotopic endometrium) on the abnormal telomere homeostasis associated with endometriosis, and on the telomere pathway in early and normal aging of the ovaries. Using VML-0501, treatment with danazol could improve the reproductive health of thousands of women with endometriosis. Clinical development is being attempted to establish a safe and effective dose of VML-0501 that can maintain ovarian function while treating the signs and symptoms of endometriosis.

[0305] Although embodiments have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided as merely examples. Those skilled in the art can conceive of numerous variations, modifications, and substitutions.

Claims

1. A method for treating a disease or condition of the endometrium in a subject, wherein the method is The subject, (a) an activator or a salt thereof; (b) 1 or more water-insoluble bioadhesives; (c) at least one oleogel; and (d) at least one aqueous gel, The procedure involves transvaginal administration of a polybioadhesive gel containing the following: Here, the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 30 times compared to oral administration of the activator or its salt to the subject; A method for administering the polybioadhesive gel via vaginal administration, thereby localizing the activator to the peritoneal tissue of the subject, and thereby treating a disease or condition of the endometrium in the subject.

2. The method according to claim 1, wherein the transvaginal administration is performed once every 24 hours.

3. The method according to claim 1, wherein the activator is a synthetic steroid.

4. The method according to claim 3, wherein the synthetic steroid is danazol or a salt thereof.

5. The method according to claim 1, wherein the activator is micronized.

6. The method according to claim 1, wherein the subject has or is suspected of having infertility.

7. The method according to claim 1, wherein the subject has or is suspected of having endometriosis.

8. The method according to claim 1, wherein the vaginal administration reduces the number of endometrial lesions in the subject by at least 10% compared to the number of endometrial lesions in the subject before the vaginal administration of the polybioadhesive gel.

9. The method according to claim 1, wherein the transvaginal administration reduces the size of one or more endometrial lesions in the subject by at least 10% compared to the size of one or more endometrial lesions before transvaginal administration of the polybioadhesive gel.

10. The method according to claim 1, wherein the polybioadhesive gel is formulated as a liquid emulsion.

11. The method according to claim 1, wherein the disease or condition of the endometrium is endometriosis, adenomyosis, or a combination thereof.

12. The method according to claim 1, wherein the polybioadhesive gel does not destroy the ovarian function of the subject.

13. The method according to claim 1, wherein the polybioadhesive gel reduces the pain in the subject by at least 10% compared to the pain identified by the subject before the polybioadhesive gel was administered transvaginally.

14. The method according to claim 1, wherein transvaginal administration of the polybioadhesive gel increases the level of the activator or salt thereof in the peritoneal tissue of the subject compared to oral administration of the activator or salt thereof to the subject.

15. The method according to claim 1, wherein the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 35 times compared to oral administration of the activator or its salt to the subject.

16. The method according to claim 1, wherein the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 40 times compared to oral administration of the activator or its salt to the subject.

17. The method according to claim 1, wherein the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 45 times compared to oral administration of the activator or its salt to the subject.

18. A method for maintaining ovarian function and reducing pain associated with endometriosis in a subject, wherein the method is: The subject, (a) an activator or a salt thereof; (b) 1 or more water-insoluble bioadhesives; (c) at least one oleogel; and (d) at least one aqueous gel The procedure involves transvaginal administration of a polybioadhesive gel containing the following: thereby maintaining ovarian function and reducing pain associated with endometriosis in the subject. Here, the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 30 times compared to oral administration of the activator or its salt to the subject; The method for administering the polybioadhesive gel via vaginal administration is to localize the activator in the peritoneal tissue of the subject.

19. The method according to claim 18, wherein the subject has, is suspected of having, or has been diagnosed with an endometrial lesion.

20. The method according to claim 18 or 19, wherein the subject has, is suspected of having, or has been diagnosed with infertility.

21. The method according to claim 18, wherein the vaginal administration is performed once every 24 hours.

22. The method according to claim 18, wherein the activator is a synthetic steroid.

23. The method according to claim 22, wherein the synthetic steroid is danazol or a salt thereof.

24. The method according to claim 18, wherein the activator is micronized.

25. The method according to claim 18, wherein the transvaginal administration reduces the number of endometrial lesions in the subject by at least 10% compared to the number of endometrial lesions in the subject before the transvaginal administration of the polybioadhesive gel.

26. The method according to claim 18, wherein the transvaginal administration reduces the size of one or more endometrial lesions in the subject by at least 10% compared to the size of one or more endometrial lesions before transvaginal administration of the polybioadhesive gel.

27. The method according to claim 18, wherein the polybioadhesive gel is formulated as a liquid emulsion.

28. The method according to claim 18, wherein the polybioadhesive gel reduces the pain in the subject by at least 10% compared to the pain identified by the subject before the polybioadhesive gel was administered transvaginally.

29. The method according to claim 18, wherein transvaginal administration of the polybioadhesive gel increases the level of the activator or salt thereof in the peritoneal tissue of the subject compared to oral administration of the activator or salt thereof to the subject.

30. The method according to claim 18, wherein the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 35 times compared to oral administration of the activator or its salt to the subject.

31. The method according to claim 18, wherein the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 40 times compared to oral administration of the activator or its salt to the subject.

32. The method according to claim 18, wherein the transvaginal administration of the polybioadhesive gel reduces the systemic concentration of the activator or its salt in the subject by at least 45 times compared to oral administration of the activator or its salt to the subject.

33. The method according to any one of claims 1 to 32, wherein, upon vaginal administration, the activator or a salt thereof diffuses into the ascites fluid of the subject, and further reaches the pelvic tissue via transvaginal absorption, thereby localizing the activator in the peritoneal cavity.

34. Any composition or method provided herein.

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