RU2713888C2 - Pharmaceutical compositions and methods based on oestradiol for intravaginal introduction - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.

SUBSTANCE: invention refers to pharmaceutical industry and represents pessary for use in treating vulvovaginal atrophy, comprising: a liquid pharmaceutical composition comprising an oestradiol and/or oestradiol hemihydrate, wherein total concentration of oestradiol and oestradiol hemihydrate in said pessary is equivalent to concentration of oestradiol in range of 1–25 mcg, where 1 mcg of oestradiol is equivalent to 1.03 mcg of oestradiol hemihydrate; and a diluting agent, which is medium chain fat; where the liquid pharmaceutical composition is encapsulated in a capsule; and where the treatment implies reducing the vaginal pH in the patient and/or involves changing the cell composition in the patient, where change in the composition of cells implies a decrease in the number of vaginal parabasal cells or an increase in the number of vaginal surface cells.

EFFECT: invention provides ease of vaginal administration, improved safety when introduced, minimum amount of vaginal discharge after administration and more effective dosage form providing improved effectiveness in treating vulvovaginal atrophy, safety and adherence of patients to treatment.

52 cl, 9 ex, 40 tbl, 14 dwg

Description

Cross references to related applications

This application claims priority over the following US patent applications: provisional US patent application Ser. No. 61/932 140 entitled "PHARMACEUTICAL COMPOSITIONS AND METHODS BASED ON ESTRADIOL FOR INTRAVAGINAL INTRODUCTION", filed January 27, 2014; provisional application for US patent Ser. No. 61/894 411 entitled "SOLUBLE CAPSULES WITH ESTRADIOL FOR INTRAVAGINAL ADMINISTRATION", filed October 22, 2013; This application is also a partial continuation of PCT / US2013 / 46443, entitled “SOLUBLE INTRAVAGINAL INTRAVAGINAL INTROVAGINAL CAPSULES CAPSULES," filed June 18, 2013, which claims priority over provisional US patent application Ser. No. 61/745 313 entitled "SOLUBLE CAPSULES WITH ESTRADIOL FOR INTRAVAGINAL INTRODUCTION", filed December 21, 2012. This application is also a partial continuation of US Ser. No. 14/099 562 entitled "NATURAL COMBINATION OF COMPOSITIONS AND METHODS OF HORMONE SUBSTITUTION THERAPY", filed December 6, 2013, which claims priority over US patent application Ser. No. 13/684 002 (now US Patent No. 8,633,178) entitled "NATURAL COMBINATION OF COMPOSITIONS AND METHODS OF HORMONE SUBSTITUTION THERAPY", filed November 21, 2012, which is a partial continuation of patent application US No. 13/843 428, entitled "NATURAL COMBINATION OF COMPOSITIONS AND METHODS OF HORMONE SUBSTITUTION THERAPY ”, filed March 15, 2013, which claims priority over provisional patent application US Ser. No. 61/661 302 entitled “PHARMACEUTICAL COMPOSITIONS BASED ON ESTRADIOL”, filed on June 18, 2012; provisional application for US patent Ser. No. 61/662 265 entitled “PHARMACEUTICAL COMPOSITIONS BASED ON PROGESTERON”, filed on June 20, 2012; and provisional patent application Ser. No. 61/563 408 entitled "NATURAL COMBINATION OF COMPOSITIONS AND METHODS OF HORMONE SUBSTITUTION THERAPY", filed November 23, 2011. All of the above applications are incorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

This application relates to pharmaceutical compositions, methods and devices that are associated with hormone replacement therapy.

Women in the postmenopausal period often suffer from atrophic vaginitis or atrophy of the vulva and vagina (hereinafter referred to as “vulvovaginal atrophy” or “BBA”) with symptoms such as vaginal dryness, vaginal odor, irritation or itching of the vagina or vulva, and urination disorders (pain, burning, or acute pain during urination), dyspareunia (pain in the vagina during intercourse), or vaginal bleeding during intercourse. Other symptoms include soreness; and frequent urination and imperative urination; urination disorder and urinary incontinence ("urination disorder (a) with estrogen deficiency"). One of the symptoms of vaginal atrophy is an increase in the pH of the vagina, which creates a more susceptible environment to infections. It was also noted that the mucosal epithelium in patients with BBA shows signs of significant atrophy, and cytological examination showed that it is accompanied by an increased number of parabasal cells and a reduced number of surface cells.

Among other things, each of these BBA-related conditions exhibits symptoms caused by a decrease in saturation of the vulvovaginal tissues with estrogen, and they can occur even in women taking oral medications based on estrogen. Although BBA most often occurs in women during menopause, the disease can occur at any point in a woman's life cycle.

Estrogen treatment has been shown to be effective in controlling the symptoms of menopause, including vaginal atrophy (BBA). Several studies have shown that symptoms associated with vaginal atrophy can often be alleviated with estrogen treatment, general or local. Existing methods of treatment have a number of problems, for example, it is a question of adherence to treatment when patients do not finish or continue treatment due to problems associated with the form of treatment.

Accordingly, this document, among other things, describes the pharmaceutical composition and dosage form of new vaginal gel capsules containing soluble estradiol for the treatment of BBA. The pharmaceutical composition of vaginal gel capsules has been developed to reduce the general limitations of other forms of estradiol for intravaginal administration. The pharmaceutical composition of the vaginal gel capsules is expected to provide ease of vaginal administration, improved safety during administration, a minimal amount of vaginal discharge after administration, and a more effective dosage form providing improved patient efficacy, safety and adherence to treatment.

SUMMARY OF THE INVENTION

In accordance with various aspects and embodiments of the present disclosure, a pharmaceutical composition of vaginal gel capsules is proposed as a potential treatment for postmenopausal women suffering from moderate or severe symptoms of BBA.

A pessary is provided herein comprising: a) a therapeutically effective amount of estradiol; and b) a medium chain fat dissolving agent.

In some embodiments, the pessary contains from about 1 μg to about 25 μg of estradiol. For example, a pessary may contain from about 1 μg to about 10 μg of estradiol; and from about 10 μg to about 25 μg of estradiol.

In some embodiments, the implementation of estradiol is soluble.

In some embodiments, the medium chain fat is at least one of a C6-C12 fatty acid or glycol, or a monoglyceride, diglyceride or triglyceride ester thereof.

In some embodiments, the solvent agent is at least one ester from the group consisting of: caproic fatty acid ester, caprylic fatty acid ester, capric fatty acid ester, and combinations thereof. For example, the dissolving agent may contain caprylic / capric acid triglyceride.

In some embodiments, the pessary is a capsule. For example, this capsule may be a soft gelatin capsule.

A pessary is also provided herein containing: a) a therapeutically effective amount of estradiol; b) caprylic / capric triglyceride; c) a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; and d) a soft gelatin capsule.

In some embodiments, the pessary as provided herein contains about 25 μg of estradiol, while administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a geometrical adjusted average maximum plasma concentration (C _max ) of estradiol of about 19 pg * h / ml to about 29 pg * h / ml; and 2) a corrected geometric mean area under the curve (AUC) of _o-24 estradiol of about 75 pg * h / ml to about 112 pg * h / ml.

In some embodiments, the pessary as provided herein contains about 25 μg of estradiol, and administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a geometric mean adjusted maximum plasma concentration (Cmax) of estrone from about 9 pg * h / ml to about 14 pg * h / ml; and 2) an adjusted geometric mean area under the curve (AUC) of 0-24 estrone ranging from about 43 pg * h / ml to about 65 pg * h / ml;

In some embodiments, the pessary as provided herein contains about 25 μg of estradiol, while administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a corrected geometric mean maximum plasma concentration (Cmax) of estrone sulfate from about 416 pg * h / ml to about 613 pg * h / ml; and 2) an adjusted geometric mean area under the curve (AUC) of 0-24 estrone sulfate, from about 3598 pg * h / ml to about 5291 pg * h / ml;

In some embodiments, the pessary as provided herein contains about 10 μg of estradiol, and administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a geometric mean adjusted maximum plasma concentration (Cmax) of estradiol of about 12 pg * h / ml to about 18 pg * h / ml; and 2) a corrected geometric mean area under the curve (AUC) of 0-24 estradiol ranging from about 42 pg * h / ml to about 63 pg * h / ml; In some embodiments, the pessary further provides a corrected geometric mean time to reach maximum plasma concentration (Tmax) of estradiol of about 1 hour to about 3 hours.

In some embodiments, the pessary as provided herein contains about 10 μg of estradiol, while administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a corrected geometric mean maximum plasma concentration (C _max ) of estrone from about 4 pg * h / ml to about 7 pg * h / ml; and 2) a corrected geometric mean area under the curve (AUC) of _0-24 estrone, from about 20 pg * h / ml to about 31 pg * h / ml; In some embodiments, the pessary additionally provides a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone from about 4 hours to about 8 hours.

In some embodiments, the pessary as provided herein contains about 10 μg of estradiol, while administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a geometrical adjusted average maximum plasma concentration (C _max ) of estrone sulfate of about 10 pg * h / ml to about 16 pg * h / ml; 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, comprising from about 56 pg * h / ml to about 84 pg * h / ml. In some embodiments, the pessary additionally provides a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone sulfate from about 4 hours to about 7 hours.

In some embodiments, the pessary as provided herein contains about 4 μg of estradiol, while administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) a geometrical adjusted average maximum plasma concentration (C _max ) of estradiol of about 4 pg * h / ml to about 8 pg * h / ml; and 2) a corrected geometric mean area under the curve (AUC) of 0-24 estradiol of about 16 pg * h / ml to about 26 pg * h / ml. In some embodiments, the pessary further provides a corrected geometric mean time to reach maximum plasma concentration (T _max ) of estradiol of about 0.25 hours to about 2 hours.

In some embodiments, the pessary as provided herein contains about 4 μg of estradiol, while administering the pessary to the patient provides (in a patient’s blood plasma sample): 1) an adjusted geometric mean maximum plasma concentration (C _max ) of estrone from about 1 pg * h / ml to about 3 pg * h / ml; 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone, from about 8 pg * h / ml to about 13 pg * h / ml; In some embodiments, the pessary additionally provides a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone from about 1 hour to about 4 hours.

In some embodiments, the pessary as provided herein contains about 4 μg of estradiol, while administering the pessary to the patient provides (in the sample

patient’s blood plasma): 1) a geometric mean adjusted maximum plasma concentration (C _max ) of estrone sulfate ranging from about 4 pg * h / ml to about 7 pg * h / ml; 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, comprising from about 22 pg * h / ml to about 34 pg * h / ml; In some embodiments, the pessary additionally provides a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone sulfate from about 1 hour to about 3 hours.

The present document also proposes a pessary containing from about 1 μg to about 25 μg of estradiol, while administering the pessary to the patient provides a corrected geometric mean maximum plasma concentration (C _max ) of estradiol that is less than about 30 pg * h / ml . For example, administering a pessary to a patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estradiol, which is less than about 18 pg * h / ml.

In some embodiments, a pessary is provided containing from about 1 μg to about 25 μg of estradiol, and administering the pessary to the patient provides a corrected geometric mean area under the (AUC) curve of 0-24 estradiol that is less than about 112 pg * h / ml . For example, administering a pessary to a patient provides an adjusted geometric mean area under the curve (AUC) of 0-24 estradiol, which is less than about 63 pg * h / ml.

In some embodiments, a pessary is provided containing from about 1 μg to about 25 μg of estradiol, wherein administering the pessary to a patient provides a corrected geometric mean maximum plasma concentration (C _max ) of estrone that is less than about 14 pg * h / ml . For example, administering a pessary to a patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone, which is less than about 7 pg * h / ml.

In some embodiments, a pessary is provided containing from about 1 μg to about 25 μg of estradiol, wherein administering the pessary to the patient provides a corrected geometric mean area under the (AUC) curve of 0-24 estrone, which is less than about 65 pg * h / ml . For example, administering a pessary to a patient provides an adjusted geometric mean area under the AUC curve of 0-24 estrone, which is less than about 31 pg * h / ml.

In some embodiments, a pessary is provided comprising from about 1 μg to about 25 μg of estradiol, and administering the pessary to the patient provides a corrected geometric mean maximum plasma concentration (C _max ) of estrone sulfate that is less than about 613 pg * h / ml For example, administering a pessary to a patient provides a corrected geometric mean of the maximum plasma concentration (C _max ) of estrone sulfate, which is less than about 16 pg * h / ml.

In some embodiments, the implementation of the proposed pessary containing from about 1 μg to about 25 μg of estradiol, while the introduction of the pessary to the patient provides a corrected geometric mean area under the curve (AUC) of 0-24 estrone sulfate, which is less than about 5291 pg * h / ml For example, administering a pessary to a patient provides an adjusted geometric mean area under the (AUC) curve of 0-24 estrone sulfate, which is less than about 84 pg * h / ml.

This document further provides a pessary containing from about 1 μg to about 25 μg of estradiol, while administering the pessary to the patient in the proximal part of the vagina provides a therapeutically effective concentration of estradiol for 24 hours in the proximal part of the vagina.

The present disclosure also provides a method for treating estrogen deficiency disorders, the method comprising administering to a patient in need of treatment a pessary, which is provided herein. In some embodiments, a method of treating vulvovaginal atrophy is provided, the method comprising administering to a patient in need of treatment a pessary, which is provided herein.

In some embodiments of the methods provided herein, treatment is about alleviating the severity of one or more symptoms taken from the group consisting of: vaginal dryness, dyspareunia, irritation of the vagina or vulva, burning of the vagina or vulva, itching of the vagina or vulva, urination disorders, and vaginal bleeding during intercourse.

In some embodiments of the methods provided herein, treatment involves lowering the pH of the vagina in a patient. For example, treatment involves lowering the pH of the vagina in a patient to a value of less than about 5.0.

In some embodiments of the methods provided herein, treatment involves changing the composition of cells in a patient. For example, a change in cell composition implies a decrease in the number of parabasal vaginal cells or an increase in the number of superficial vaginal cells. In some embodiments, the decrease in the number of parabasal vaginal cells in a patient is at least about 35% (eg, at least about 50%). In some embodiments, the increase in the number of surface vaginal cells is at least about 5% (for example, at least about 35%).

This document further provides a method for reducing vaginal discharge after administration of the pessary, the method involves administering a pessary to the patient in need of treatment, which is provided herein, wherein vaginal discharge after administration of the pessary is compared with vaginal discharge after administration of the reference drug.

BLUEPRINTS

The aforementioned features and objectives of the disclosure of the essence of the present invention will become clearer when reading the following description, considered in conjunction with the accompanying drawings, in which like reference numbers indicate like elements, and where:

in FIG. 1 is a diagram depicting a process in accordance with various embodiments of the present invention;

in FIG. 2 shows a suppository in accordance with various embodiments of the present invention.

in FIG. Figure 3 shows a linear diagram of the average plasma concentrations of estradiol - the dependence of the concentrations on time, adjusted for the initial values (N = 36);

in FIG. Figure 4 shows a graph on a semi-logarithmic scale of the average concentrations of estradiol in blood plasma - the dependence of the concentrations on time, adjusted for the initial values (N = 36);

in FIG. Figure 5 presents a linear diagram of the average concentrations of estrone in the blood plasma - the dependence of the concentrations on time adjusted for the initial values (N = 36);

in FIG. Figure 6 shows a graph on a semi-logarithmic scale of the average concentrations of estrone in plasma - the dependence of the concentrations on time adjusted for the initial values (N = 36);

in FIG. Figure 7 presents a linear diagram of the average plasma concentrations of estrone sulfate in blood - the concentration versus time dependence adjusted for the initial values (N = 36);

in FIG. Figure 8 shows a graph on a semi-logarithmic scale of the average plasma concentrations of estrone sulfate in blood - the concentration dependence of time on time adjusted for the initial values (N = 36);

in FIG. Figure 9 shows a linear diagram of the average plasma concentrations of estradiol - the concentration as a function of time, adjusted for the initial values (N = 34);

in FIG. 10 shows a graph on a semi-logarithmic scale of the average concentrations of estradiol in blood plasma - the dependence of the concentrations on time, adjusted for the initial values (N = 34);

in FIG. 11 is a linear diagram of the average plasma concentrations of estrone — the concentration as a function of time versus initial values (N = 33);

in FIG. 12 shows a graph on a semi-logarithmic scale of the average concentrations of estrone in blood plasma - the dependence of the concentrations on time adjusted for the initial values (N = 33);

in FIG. Figure 13 shows a linear diagram of the average plasma concentrations of estrone sulfate in blood - the concentration versus time dependence adjusted for the initial values (N = 24); and

in FIG. 14 shows a graph on a semi-logarithmic scale of the average concentrations of estrone sulfate in blood plasma - the dependence of the concentrations on time, adjusted for the initial values (N = 24);

DETAILED DESCRIPTION

In the following detailed description of embodiments, as part of the disclosure of the present invention, reference is made to the accompanying drawings, in which like numbers refer to like elements, and where, by way of example, specific embodiments are given that can be implemented by the present disclosure. These embodiments are explained in sufficient detail to enable those skilled in the art to implement the present disclosure, and it should also be understood that other embodiments may be used and other changes may be made without departing from the scope of the present disclosure. Thus, the following detailed description should not be taken as limiting, and the scope of the present disclosure is determined solely by the attached claims. It should be understood that the term “or” used in this disclosure means a logical disjunction (that is, and / or), and not excluding a disjunction, unless specifically indicated using terms such as “one of two”, “unless "," Alternatively "or other words with the same meaning.

NOTATION

As used herein, the term “active pharmaceutical ingredient” (“API”) refers to the active compound (s) contained in a drug.

As used herein, the term “co-administered” means that two or more drugs are administered simultaneously or sequentially for one or different days.

As used herein, the term “drug” means at least one active pharmaceutical ingredient in combination with at least one excipient in the form of a unit dosage form.

The term "area under the curve"("AUC") refers to the area under the curve limited by changes in blood concentrations of the active pharmaceutical ingredient (eg, estradiol or progesterone) or the metabolite of the active pharmaceutical ingredient over time after a dose of the active pharmaceutical ingredient is administered. “AUC _o-∞ ” is the area under the concentration-time curve extrapolated to infinity after a dose of the drug is administered. “AUC _ot ” is the area under the concentration-time curve from zero to time t after the dose of the drug is administered, and t is the last time point with the measured concentration.

The term "C _max " refers to the maximum value of the concentration in the blood shown on the curve, which reflects the changes in the blood concentration of the active pharmaceutical ingredient (eg, progesterone or estradiol) or the metabolite of the active pharmaceutical ingredient over time.

The term "T _max " refers to the time it takes an active pharmaceutical ingredient (eg, estradiol or progesterone) or a metabolite of an active pharmaceutical ingredient to achieve maximum blood concentration.

The term “bioavailability”, the meaning of which is defined in § 320.1 (a) 21 of the Code of Federal Regulations, refers to the rate and extent of absorption of APIs, an active ingredient or an active substance from a drug and getting into the place of application of the action. For example, bioavailability can be defined as the amount of API in the blood (serum or plasma) versus time. Pharmacokinetic (PK) parameters such as AUC, C _max or T _max can be used to measure and evaluate bioavailability. For drugs that are not intended to enter the bloodstream, bioavailability can be assessed using parameters designed to display the rate and extent to which the API, active ingredient, or active substance reaches the site of application of the action.

The term “bioequivalent”, the meaning of which is defined in § 320.1 (e) 21 of the Code of Federal Regulations, refers to the absence of a significant difference in the rate and extent of entry of APIs, active ingredient or active substance in pharmaceutical equivalents or pharmaceutical alternatives at the site of application of the drug, if they were administered in the same molar dose under similar conditions in a suitably designed study. If there is a deliberate rate difference (for example, in some sustained release dosage forms), some pharmaceutical equivalents or alternatives may be considered bioequivalents unless there is a significant difference in the extent to which the active ingredient or substance of each drug reaches the site of application of the drug. This is applicable only if the difference in speed at which the active ingredient or substance gets into the place of application of the drug is intentional and displayed in the intended labeling is not significant to achieve effective concentrations of the drug in the body with continuous use and is considered insignificant for the drug from a medical point of view. In practice, two drugs are considered bioequivalent if the confidence interval of 90% for AUC, C _max or, optionally, T _max is in the range from 80% to 125%.

The terms “bioidentical,” “identical to natural,” or “natural,” as used in relation to the hormones described herein, refer to hormones that correspond in chemical structure and action to hormones that are naturally or endogenously produced in the human body. An example of natural estrogen is estradiol.

The term “bioidentical hormone” or “natural identical hormone” refers to an active pharmaceutical ingredient whose structure is identical to a hormone that is naturally or endogenously produced in the human body (for example, estradiol and progesterone).

The term “estradiol” refers to a (17β) -estra-1,3,5 (10) -trien-3,17-diol. Estradiol is also interchangeably called 17β-estradiol, estradiol or E2 and is produced endogenously in the human body. In this document, estradiol refers to a bioidentical or identical natural form of estradiol, which is produced in the human body and has the following structure:

Estradiol is produced in anhydrous or hemihydrate form. In order to disclose the essence of the present invention, the anhydrous form or the form of the hemihydrate can replace each other, due to the presence of water or its lack, in accordance with well-known and understandable methods.

The term “soluble estradiol” means that estradiol or a portion thereof is soluble or can be dissolved in the dissolving agent (s) or compositions described herein. Soluble estradiol may be estradiol, which is approximately 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% soluble. In some embodiments, the estradiol is “completely soluble,” wherein all or substantially all of the estradiol is soluble or can be dissolved in a dissolving agent. Fully soluble estradiol may be estradiol, which is soluble by approximately 97%, 98%, 99% or 100%. Solubility can be expressed as a mass fraction (% w / w, which is also indicated as% w / w).

The term "progesterone" refers to pregn-4-en-3,20-dione. Progesterone is also interchangeably called P4 and is produced endogenously in the human body. In this document, progesterone refers to the bioidentical or identical natural form of progesterone that is produced in the human body and has the following structure:

The term “soluble progesterone” means that progesterone or a part thereof is soluble or can be dissolved in the dissolving agent (s) or compositions described herein. In some embodiments, the progesterone is “partially soluble,” wherein part of the progesterone is soluble or can be dissolved in the dissolving agent, and part of the progesterone is suspended in the dissolving agent. Partially soluble progesterone can be progesterone, which is soluble by approximately 1%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 85%, 90% or 95%. In other embodiments, the progesterone is “completely soluble,” wherein all or substantially all of the progesterone is soluble or can be dissolved in a dissolving agent. Fully soluble progesterone can be progesterone, which is soluble by approximately 97%, 98%, 99% or 100%. Solubility can be expressed as a mass fraction (% w / w, which is also indicated as% w / w).

As used herein, the terms “micronized progesterone” and “micronized estradiol” mean micronized progesterone and micronized estradiol with an X50 particle size of less than about 15 microns or an X90 particle size of less than about 25 microns. The term "X50" means that the diameter of half the particles of the sample is less than a given number. For example, micronized progesterone with an X50 of 5 microns means that in this micronized progesterone sample, half the particle diameter is less than 5 microns. Similarly, the term “X90” means that the diameter of ninety percent (90%) of the sample particles is less than a predetermined number.

The term “glyceride” means a glycerol ester (1,2,3-propanetriol) with acyl radicals of fatty acids, also known as acylglycerol. If only one position of the glycerol molecule is esterified with a fatty acid, a “monoglyceride” or “monoacylglycerol” is obtained; if two positions are esterified, “diglyceride” or “diacylglycerol” is obtained; and if the three positions of the glycerol molecule are esterified with a fatty acid, the result is “triglyceride” or “triacylglycerol”. A glyceride is “simple” if all the esterified sites contain the same fatty acid; in turn, the glyceride is “mixed” if the esterified sites contain different fatty acids. The carbon atoms of the main chain of the glycerol molecule are denoted by sn-1, sn-2 and sn-3, with sn-2 located in the center and sn-1 and sn-3 at the edges of the main chain of the glycerol molecule.

The term “dissolving agent” refers to an agent or combination of agents that dissolve an active pharmaceutical ingredient (eg, estradiol or progesterone). Suitable dissolving agents are unlimitedly represented, for example, by medium chain fats, as well as other solvents and cosolvents that dissolve the active pharmaceutical ingredient to the desired degree. Solvents suitable for use in the compositions described herein are pharmaceutical grade solvents (e.g. pharmaceutical grade medium chain fats). It will be apparent to those skilled in the art that other adjuvants or components can be added to or mixed with the solvent improving the practical properties of the dissolving agent or the resulting composition. Such excipients are unlimitedly represented, for example, by surfactants, emulsifiers, thickeners, colorants, flavors, etc. In some embodiments, the dissolving agent is a medium chain fat, and in some embodiments, the medium chain fat is combined with a co-solvent (s) or other auxiliary agent (s) (s).

The term “medium chain” is used to describe the length of the aliphatic chain in molecules containing fatty acids. The term “medium chain” in particular refers to fatty acids, fatty acid esters or fatty acid derivatives that contain aliphatic fatty acid tails or carbon chains that contain from 6 (C6) to 14 (C14) carbon atoms, from 8 (C8) to 12 (C12) carbon atoms or from 8 (C8) to 10 (C10) carbon atoms.

The terms “medium chain fatty acid” and “medium chain fatty acid derivative” are used to describe fatty acids or derivatives of fatty acids with aliphatic tails (i.e., carbon chains) that contain from 6 to 14 carbon atoms. Fatty acids contain a straight or branched aliphatic tail attached to the carboxylic acid functional group. Derivatives of fatty acids are, for example, fatty acid esters and molecules containing fatty acids, unlimitedly represented by mono-, di- and triglycerides, which contain components derived from fatty acids. Derivatives of fatty acids are also ethylene or propylene glycol esters of fatty acids. Aliphatic tails may be saturated or unsaturated (i.e., have one or more double bonds between carbon atoms). In some embodiments, aliphatic tails are saturated (i.e., do not have double bonds between carbon atoms). Medium chain fatty acids or derivatives of medium chain fatty acids include aliphatic tail chains that have 6 to 14 carbon atoms, including C6-C14, C6-C12, C8-C14, C8-C12, C6-C10, C8-C10 chains or other. Medium chain fatty acids are unlimitedly represented, for example, by caproic acid, caprylic acid, capric acid, lauric acid, myristic acid and their derivatives.

As used herein, the term “fat” refers to any pharmaceutically acceptable fat, in particular medium chain fats, in particular excluding peanut butter, which can provide the suspension or dissolution of bioidentical progesterone or estradiol, including their starting materials or precursors, including including micronized progesterone or micronized estradiol, as described herein.

The term "medium chain fat" refers to fat, wherein the composition of the fatty acid fraction in that fat is essentially medium chain fatty acids (i.e., C6 to C14), i.e. the fatty acid profile in fat is essentially a medium chain. As used herein, the term “essentially” means that from 20% to 100% (including the upper and lower limits) of the fatty acid fraction in the fat consists of medium chain fatty acids, i.e. fatty acids with aliphatic tails (i.e. carbon chains) that contain from 6 to 14 carbon atoms. In some embodiments, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 85 %, about 90% or about 95% of the fatty acid fraction in the fat consists of medium chain fatty acids. It will be readily apparent to those skilled in the art that the terms “alkyl content” and “alkyl distribution” in fat may be used in place of the term “fatty acid fraction” in fat to describe this fat or dissolving agent, and that these terms are used interchangeably herein . In this regard, medium chain fats suitable for use in the formulations described herein are medium chain fats in which the fatty acid fraction in the fat is essentially medium chain fatty acids or medium chain fats in which the alkyl content or distribution of the alkyl in the fat is essentially medium chain alkyls (C6-C12 alkyls). Those skilled in the art will understand that medium chain fats suitable for use in the formulations described herein have a pharmaceutical category (e.g., pharmaceutical chain medium chain fats). Medium chain fats are unboundedly represented, for example, by medium chain fatty acids, glycerol esters of medium chain fatty acids (e.g. mono-, di- and triglycerides), propylene glycol ethers of medium chain fatty acids, propylene glycol derivatives of medium chain fatty acids, and combinations thereof.

The term "ELF" or "equivalent carbon number" means the sum of the number of carbon atoms in the fatty acid chains in fat and can be used to describe fat such as, for example, medium chain fat or long chain fat. For example, the EEC of tripalmitin (tripalmitic glycerol), which is a simple triglyceride containing three chains of fatty acids of 16 carbon atoms, is 3 × 16 = 48. Conversely, a triglyceride with an ECH = 40 may contain a “mixed” chain length of fatty acids of 8, 16 and 16 atoms; 10, 14 and 16 atoms; 8, 14 and 18 atoms; etc. Naturally occurring fats are often “mixed” with respect to specific fatty acids, however, as a rule, they do not contain either long chain fatty acids or medium chain fatty acids in the same main chain of the glycerol molecule. Thus, triglycerides with an EEC from 21 to 42 usually contain predominantly medium chain fatty acids; while triglycerides with EECs above 43 usually contain predominantly long chain fatty acids. For example, the EOF of corn oil triglyceride in F. USA will range from 51 to 54. Medium-chain diglycerides with EECs from 12 to 28 will often contain predominantly chains of medium-chain fatty acids, while diglycerides with EECs of 32 or more will typically , contain predominantly tails of long chain fatty acids. Monoglycerides will have an EEC that matches the chain length of a single chain fatty acid. Thus, monoglycerides with an EEC in the range from 6 to 14 contain predominantly medium chain fatty acids, and monoglycerides with an EEC 16 or more will contain predominantly long chain fatty acids.

The average EEC value of medium chain triglyceride fats is usually from 21 to 42. For example, as listed in the US Pharmacopoeia (US F.), medium chain triglycerides have a composition similar to the fat shown in the table below:

Fatty Acid Tail Length % fat Fat example 6 < 2.0 2.0 8 50.0-80.0 70.0 10 20.0-50.0 25.0 12 < 3.0 2.0 14 < 1.0 1,0

and will have an average value of the ECH 3 * [(6 * 0.02) + (8 * 0.70) + (10 * 0.25) + (12 * 0.02) + (14 * 0.01)] = 25.8. The EEC of medium chain triglyceride fat, given as an example, can also be represented as a range (in the ranges established in the US F.) from 24.9 to 27.0. For fats containing mixed mono-, di, and triglycerides, or for single and double bond fatty acid glycols, the total fat of EFC can be determined by calculating the EFC of each individual component (for example, C8 monoglycerides, C8 diglycerides, C10 monoglycerides and C10 monoglycerides) and taking the sum of the relative percentage value of the component, multiplied by the ECH, normalized with monoglyceride for each component. For example, a fat containing C8 and C10 mono- and diglycerides, as shown in the table below, has an ELF of 8.3, and thus is a medium chain fat.

Fatty Acid Chain Length % fat ESF as% fat (chain length) × (% in
fat) ESF as% fat normalized with monoglyceride C8 monoglyceride 47 8 × 0.47 = 3.76 3.76 C10 monoglyceride 8 10 × 0.08 = 0.8 0.8 C8 diglyceride 38 2 × (8 × 0.38) = 6.08 6.08 / 2 = 3.04 C10 diglyceride 7 2 × (10 × 0.07) = 1.4 1.4 / 2 = 0.7 Eouch fat
(normalized with monoglyceride) 8.3

In other words, the ECF can be calculated by multiplying each chain length in the composition by its relative percentage in fat: (8 * 0.85) + (10 * 0.15) = 8.3.

As used herein, the term “excipients” refers to inactive pharmaceutical ingredients, such as dissolving agents, antioxidants, oils, lubricants, and other substances included in pharmaceutical products.

The term “patient” or “subject” refers to a person to whom the pharmaceutical composition is administered.

The term "pharmaceutical composition" refers to a pharmaceutical composition containing at least a dissolving agent and estradiol. In this document, pharmaceutical formulations are administered, for example, in the form of a pessary (i.e., a vaginal suppository) or are absorbed vaginally.

The term "progestin" means any natural or human-made substance that has pharmacological properties similar to those of progesterone.

The term “reference drug” (“LPS”) refers to VAGIFEM® (vaginal tablets with estradiol) or to ESTRACE® vaginal cream.

The terms “treat,” “treat,” and “treatment,” refer to any signs of success in treating or repairing an injury, illness, or condition, including any objective or subjective indicator, such as a reduction in pain or the severity of a symptom; remission; relieving symptoms or improving patient tolerance of damage, illness, or condition; slowing down degenerative processes or worsening; or improving the physical and mental condition of the patient. Treatment or elimination of symptoms may be based on objective or subjective indicators, including the results of a physical examination, neuropsychiatric examinations, or psychiatric examination.

The terms “atrophic vaginitis”, “vulvovaginal atrophy”, “vaginal atrophy” and “BBA” are used interchangeably herein. The molecular morphology of VVA is well known in the medical field.

INTRODUCTION

Pharmaceutical formulations containing soluble estradiol formulated for absorption by the vaginal route are provided herein. The pharmaceutical compositions described herein are designed to absorb and provide a local therapeutic effect, for example, in vaginal or surrounding tissues. Additionally, data are provided herein that demonstrate the effectiveness of the described pharmaceutical formulations as well as methods related to pharmaceutical formulations. In general, the pharmaceutical formulations described herein are effective in treating BBA, dyspareunia, and other symptoms caused by a decrease in estrogen or a lack thereof.

Additional aspects and embodiments of the disclosure of the essence of the present invention include: providing greater ease of use of the drug by the patient, thereby potentially minimizing some side effects caused by improper administration, minimizing the incidence of vulvovaginal fungal infections in comparison with the frequency of vulvovaginal fungal infections arising from the use of other estradiol-based preparations used vaginally; and reduced side effects (eg, itching) compared with the reference drug: VAGIFEM® (estradiol-based vaginal tablets, Novo Nordisk; Princeton, NJ).

PHARMACEUTICAL COMPOSITION

Functional properties

In accordance with embodiments, the pharmaceutical compositions described herein are alcohol free or substantially free of alcohol. Pharmaceutical formulations provide improved patient adherence due to improvements in the prior art. In accordance with embodiments, the pharmaceutical compositions described herein are encapsulated in a soft gelatin capsule, which provides greater comfort during use. In accordance with embodiments, the pharmaceutical compositions are substantially liquid, so they are better absorbed by the vaginal tissue and dispersed over a larger surface area of the vaginal tissue.

Estradiol

In accordance with embodiments, the pharmaceutical compositions described herein are intended for intravaginal administration in a single or multiple unit dosage form. In accordance with embodiments, estradiol in pharmaceutical formulations is at least about: 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85% , 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% soluble. According to embodiments in which estradiol is not 100% soluble, the remainder of estradiol is present in a micronized (crystallized) form that is absorbed in the body and retains biological functional properties either in its micronized form or in another form into which it is converted micronized form after administration.

According to embodiments, all or part of estradiol can be dissolved in a dissolving agent during the manufacturing process. In accordance with embodiments, all or part of estradiol is dissolved after administration (for example, the micronized part, in which estradiol is 100% insoluble, is dissolved in body fluid after administration). In accordance with embodiments, due to the fact that estradiol is soluble, the dissolving agents mentioned herein, together with additional excipients other than or without dissolving agents, may be liquid or semi-liquid. To the extent that estradiol is not completely soluble at the time of administration / use, estradiol should be essentially soluble at body temperature (average 37 ° C) and, in general, at vaginal pH (value varies from 3.8 to 4.5 in healthy patients; and from 4.6 to 6.5 in patients with BBA).

In accordance with embodiments, estradiol may be added to the pharmaceutical compositions described herein in the form of estradiol, estradiol hemihydrate, or other forms of estradiol used in pharmaceutical compositions or dosage forms.

In accordance with embodiments, dosages of estradiol may vary. The dosage of estradiol (or estradiol hemihydrate, for example, to the extent that the water content of estradiol hemihydrate is taken into account) is at least about 1 microgram (μg) to about 50 μg. In specific embodiments, the dosage is at least about: 1 μg, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, 7 μg, 8 μg, 9 μg, 10 μg, 11 μg, 12 μg, 13 mcg, 14 mcg, 15 mcg, 16 mcg, 17 mcg, 18 mcg, 19 mcg, 20 mcg, 21 mcg, 22 mcg, 23 mcg, 24 mcg, 25 mcg, 26 mcg, 27 mcg, 28 mcg, 29 mcg, 30 mcg, 31 mcg, 32 mcg, 33 mcg, 34 mcg, 35 mcg, 36 mcg, 37 mcg, 38 mcg, 39 mcg, 40 mcg, 41 mcg, 42 mcg, 43 mcg, 44 mcg, 45 mcg, 46 mcg , 47 mcg, 48 mcg, 49 mcg or 50 mcg estradiol. In accordance with embodiments, the pharmaceutical compositions contain at least about 2.5 μg; 4 μg, 6.25 μg, 7.5 μg, 12.5 μg, 18.75 μg of estradiol. In accordance with embodiments, the pharmaceutical compositions contain from about 1 μg to about 10 μg, from 3 μg to 7 μg, from about 7.5 μg to 12.5 μg, from about 10 μg to about 25 μg, about 1 μg, about 2.5 μg, from about 23.5 μg to 27.5 μg, from about 7.5 μg to 22.5 μg, from 10 μg to 25 μg of estradiol. The lowest clinically effective dose of estradiol is used to treat BBA and other symptoms described in this document. In some embodiments, the dose of estradiol is about 4 μg. In one embodiment, the dose of estradiol is about 10 μg. In another embodiment, the dose of estradiol is about 25 μg.

Solvent system

In accordance with embodiments, a solvent system that dissolves estradiol is a medium chain fatty acid solvent along with other excipients. In accordance with embodiments, the solvent system is non-toxic, pharmaceutically acceptable solvents, cosolvents, surfactants, and other excipients suitable for intravaginal administration or absorption.

In accordance with embodiments, fats, the predominant component of which are medium chain fatty acids, are used as dissolving agents to dissolve estradiol. In accordance with embodiments, the dissolving agents are medium chain fatty acid esters (e.g., glycerol, ethylene glycol or propylene glycol esters) or mixtures thereof. In accordance with embodiments, the medium chain fatty acids comprise chains of length C6 to C14. In accordance with embodiments, the medium chain fatty acids comprise chains of length C6 to C12. In accordance with embodiments, the medium chain fatty acids comprise essentially chains of lengths C8 to C10. The ECH of medium chain fats will range from 21 to 42 for triglycerides, from 12 to 28 for diglycerides and from 6 to 14 for monoglycerides.

In accordance with embodiments, mid chain fatty acids are saturated. In accordance with embodiments, medium chain fatty acids are predominantly saturated, i.e. more than about 60% or more than about 75%.

In accordance with embodiments, estradiol is dissolved in a dissolving agent at room temperature, although it may be desirable for some dissolving agents to be heated during production to improve viscosity. In accordance with embodiments, the dissolving agent will be liquid at a temperature in the range of from room temperature to about 50 ° C, at 50 ° C or lower, at 40 ° C or lower, or at 30 ° C or lower.

In accordance with embodiments, the solubility of estradiol in a medium chain fat, medium chain fatty acid, or solvent (or fat / surfactant) is at least about 0.01% by weight, 0.02% by weight, 0.05 % wt., 0.06% wt., 0.08% wt., 0.1% wt., 0.2% wt., 0.3% wt., 0.4% wt., 0.5% weight., 0.6% weight., 0.7% weight., 0.8% weight., 0.9% weight., 1.0% weight.

In accordance with embodiments, the medium chain dissolving agents are unlimitedly represented, for example, by saturated medium chain fatty acids: caproic acid (C6), enanthic acid (C7), caprylic acid (C8), pelargonic acid (C9), capric acid (C10), undecyl acid (C11), lauric acid (C12), tridecanoic acid (C13) or myristic acid (C14). In accordance with embodiments, the dissolving agent is fats consisting of these free medium chain fatty acids, glycerol, propylene glycol or ethylene glycol esters of medium chain fatty acids, or combinations thereof. These examples advantageously comprise saturated medium chain fatty acids (i.e., more than 50% of the fatty acids are medium chain saturated fatty acids). In accordance with the options for implementation are predominantly saturated C6-C12 fatty acids. In accordance with embodiments, the solvent agent is selected from at least one of a solvent and a co-solvent.

In accordance with embodiments, glycerol-based dissolving agents comprise: mono-, di-, or triglycerides, and combinations thereof and derivatives thereof. Examples of glycerol-based solvent agents include MIGLYOL® preparations, which are caprylic / capric acid triglycerides (SASOL Germany GMBH, Hamburg). MIGLYOL preparations include MIGLYOL 810 (caprylic / capric acid triglyceride), MIGLYOL 812 (caprylic / capric acid triglyceride), MIGLYOL 816 (caprylic / capric acid triglyceride) and MIGLYOL 829 (capric / capric acid triglyceride). Similarly, other solvent agents with caprylic / capric acid triglycerides are meant, including, for example, caproic / caprylic / capric / lauric acid triglycerides; caprylic / capric / linoleic acid triglycerides; caprylic / capric / succinic acid triglycerides. In accordance with embodiments of the CAPMUL MCM, medium chain mono- and diglycerides are a dissolving agent. Other fatty acid triglycerides from fractionated vegetable oils and combinations or derivatives thereof may be a dissolving agent in accordance with embodiments. For example, the solvent may be 1,2,3-propanetirol (glycerol, glycerol) esters of saturated coconut and palm kernel oils and their derivatives.

Solvents based on ethylene and propylene glycol (which contain polyethylene and polypropylene glycols) include: glyceryl mono- and dicaprilates; propylene glycol monocaprylate (e.g., CAPMUL® PG-8 (CAPMUL brands are owned by ABITEC Columbus, Ohio)); propylene glycol monocaprate (e.g., CAPMUL PG-10); propylene glycol mono- and dicaprilates; propylene glycol mono- and dicaprate; diethylene glycol monoester (e.g. TRANSCUTOL®, 2- (2-ethoxyethoxy) ethanol, GATTEFOSSÉ SAS); diethylene glycol monoethyl ether. Other combinations of mono- and diesters of propylene glycol or ethylene glycol, expressly implied, are a dissolving agent.

In accordance with embodiments, the dissolving agent is a combination of mono- and di-propylene and ethylene glycols, as well as combinations of mono, di, and triglycerides. In accordance with embodiments, polyethylene glycol glyceride (GELUCIRE®, GATIEFOSSÉ SAS, Saint-Prix, France) may be used herein as a solvent or surfactant. For example, GELUCIRE 44/14 (PEG-32 glyceryl laurate, Heb. F.), medium chain fatty acid polyethylene glycol esters, is a polyethylene glycol glyceride consisting of mono-, di- and triglycerides, as well as mono- and polyethylene glycol diesters.

In accordance with embodiments, commercially available dissolving agents based on glycerol and glycol fatty acid esters are often derived from natural fats, therefore, in addition to fatty acid esters, they can contain components that primarily constitute and characterize the dissolving agent. Similar other components may be, for example, mono-, di- and triglycerides of other fatty acids; ethylene glycol or propylene glycol mono- and diesters of fatty acids, free glycerols, free glycols or free fatty acids. In some embodiments, the implementation, if the fat / dissolving agent described herein is a glycerol mono- or diester of a saturated C8 fatty acid, the main component of fat, which is> 50% by weight. (e.g.,> 75% wt.,> 85% wt. or> 90% wt.) are caprylic acid monoglycerides and caprylic acid diglycerides. For example, the ABITEC specification for CAPMUL MCM C8 indicates that CAPMUL MCM C8 consists of medium chain fatty acid mono- and diglycerides (mainly caprylic) and indicates that the alkyl content is: ≤1% C6, ≥95% C8, ≤5 % C10 and ≤1.5% C12 or more.

For example, MIGLYOL 812 is a dissolving agent, which is usually described as a C8-C10 triglyceride, since the composition of the fatty acids is as follows: at least about 80% triglyceride esters of caprylic acid (C8) and capric acid (C10). However, it also contains small amounts of other fatty acids, for example, less than about 5% caproic acid (C6), lauric acid (C12) and myristic acid (C14). In the specification for the various MIGLYOL preparations, the various components of the fatty acids are presented as follows:

Research 810 812 818 829 840 Caproic acid (C6: 0) Max. 2.0 Max. 2.0 Max. 2 Max. 2 Max. 2 Caproic acid (C8: 0) 65.0-80.0 50.0-65.0 45-65 45-55 65-80 Caproic acid (C10: 0) 20.0-35.0 30.0-45.0 30-45 30-40 20-35 Lauric acid (C12: 0) Max. 2 Max. 2 Max. 3 Max. 3 Max. 2 Myristic acid (C14: 0) Max. 1,0 Max. 1,0 Max. 1 Max. 1 Max. 1 Linoleic acid (18: 2) - - 2-5 - - succinic acid - - - 15-20 - EUCH 25.5-26.4 26.1-27 26.52-28.56 26-27.6 25.5-26.4

In accordance with embodiments, anionic or nonionic surfactants may be used in pharmaceutical compositions containing soluble estradiol. The ratios of the solvent (s) of the agent (s) to the surface-active (s) substance (s) vary depending on the respective (their) solvent (s) agent (s) and the corresponding (s) surface-active substance (s) (in ) and the necessary physical properties of the obtained pharmaceutical composition. By way of non-limiting example, CAPMUL MCM and a non-ionic surfactant can be used in relationships such as 65:35, 70:30, 75:25, 80:20, 85:15 and 90:10. Other non-limiting examples imply that: CAPMUL MCM and GELUCIRE 39/01 are used as a non-limiting example in relationships such as 6: 4, 7: 3, and 8: 2; CAPMUL MCM and GELUCIRE 43/01 are used as a non-limiting example in such ratios as 7: 3 and 8: 2; CAPMUL MCM and GELUCIRE 50/13 are used as a non-limiting example in such ratios as 7: 3, 8: 2 and 9: 1.

Other excipients

In accordance with embodiments, the pharmaceutical composition further comprises a surfactant. The surfactant may be a non-ionic surfactant, a cationic surfactant, an anionic surfactant, or mixtures thereof. Suitable surfactants are, for example, water-insoluble surfactants having a hydrophilic lipophilic balance (HLB) of less than 12, and water-soluble surfactants having a HLB of more than 12. Surfactants, which have a high HLB value, contribute to the formation of oil-water drops. Surfactants are amphiphilic in nature and can dissolve relatively large amounts of hydrophobic drug compounds.

Non-limiting examples include: tween, dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), ethanol, glycerol, N-methyl-2-pyrrolidone (NMP), PEG 300, PEG 400, poloxamer 407, propylene glycol, phospholipids, hydrogenated soybean phosphatidylcholine ( , distearoyl-phosphatidylglycerol (DSPH), L-α-dimyristoyl-phosphatidylcholine (DMPC), L-α-dimyristoyl phosphatidylglycerol (DMF), polyoxyl 35 castor oil (CREMOPHOR EL, CREMOPHOR ELECTROGEN 40 Oil ), Polyoxyl 60 hydrogenated castor oil (CREMOPHOR RH 60), Polysorbate 20 (TWE EN 20), polysorbate 80 (TWEEN 80), d-α-tocopherol polyethylene glycol 1000 succinate (TPGS), solutol HS-15, sorbitan monooleate (SPAN 20), PEG 300 caprylic / capric acid glycerides (SOFTIGEN 767), PEG 400 glycerides caprylic / capric acid (LABRASOL), PEG 300 oleic acid glycerides (LABRAFIL M-1944CS), castor oil polyoxyl 35 (ETOCAS 35), caprylate glyceryl (mono- and diglycerides) (IMWITOR), PEG 300 linoleic acid glycerides MRA-LAB (LAB 2125CS), polyoxyl 8 stearate (PEG 400 monostearate), polyoxyl 40 stearate (PEG 1750 monostearate), and combinations thereof. Further suitable surfactants include, for example, polyoxyethylene sorbitan monolaurate derivatives such as polysorbate, capryl caproyl macrogol glycerides, polyglycolized glycerides, and the like.

In accordance with embodiments, the nonionic surfactant is selected from one or more long chain fatty acid glycerol and polyethylene glycol esters, for example, lauroylmacrogol-32 glycerides or lauroylpolyoxyl-32 glycerides, commercially available under the name GELUCIRE, including, for example, GELUCIRE 39 / 01 (glycerol esters of saturated C12-C18 fatty acids), GELUCIRE 43/01 (solid fat from NF / JPE (National Pharmacopoeia Formulary / Excipient Collection in the Japanese Pharmacopoeia)) and GELUCIRE 50/13 (stearoy glycerides macrogol-32 (Eur. F.), glycerides stearoilpolioksila-32 NF, stearoilpolioksilglitseridy (FDA IIG USA)). Such surfactants can be used in concentrations in excess of about 0.01%, and are usually contained in various amounts of about 0.01% -10.0%, 10.1% -20% and 20.1% -30%. In some embodiments, surfactants can be used in concentrations from about 1% to about 10% (for example, from about 1% to about 5%, from about 2% to about 4%, from about 3% to about 8%) .

In some embodiments, non-ionic surfactants are unlimitedly represented, for example, by one or more of oleic acid, linoleic acid, palmitic acid, and stearic acid. In accordance with embodiments of nonionic surfactants are polyethylene sorbitan esters including polysorbate 80, which is commercially available under the tradename TWEEN ^® 80 (polysorbate 80) (Sigma Aldrich, St. Louis, Missouri). Polysorbate 80 contains approximately 60% -70% oleic acid, the remainder mainly contains linoleic acids, palmitic acids and stearic acids. Polysorbate 80 can be used in amounts ranging from about 5 to 50%, and in accordance with some embodiments, comprise about 30% of the total weight of the pharmaceutical composition.

In accordance with embodiments of the nonionic surfactant is PEG-6 palmitostearate and ethylene glycol palmitostearate, which are commercially available under the name TEFOSE ^® 63 (GATTEFOSSÉ SAS, Saint-Prix, France), which can be used, for example, CAPMUL MCM from the ratio of MCM to TEFOSE 63, for example, 8: 2 or 9: 1. In accordance with embodiments, combinations of other solvents / nonionic surfactants are, for example, MIGLYOL 812: GELUCIRE 50/13 and MIGLYOL 812: TEFOSE 63.

In accordance with embodiments, the surfactants may be anionic surfactants, for example: ammonium lauryl sulfate, sodium dioctyl sulfosuccinate, perfluorooctanesulfonic acid, potassium lauryl sulfate, or sodium stearate. Cationic surfactants are also considered.

In accordance with embodiments, nonionic or anionic surfactants may be used alone with at least one solvent or in combination with other surfactants. Accordingly, such surfactants or any other adjuvant described herein can be used to dissolve estradiol. A combination of a dissolving agent, a surfactant, and other excipients with which estradiol will be absorbed by the vaginal tissue should be developed. In accordance with embodiments, the pharmaceutical composition will result in a minimal amount of vaginal discharge.

In accordance with embodiments, the pharmaceutical composition further comprises at least one thickening agent. In general, a thickening agent is added if the viscosity of the pharmaceutical composition causes a degree of absorption worse than desired. In accordance with embodiments, the surfactant (s) described herein can also provide a thickening of the pharmaceutical composition so that after release, estradiol can be more easily absorbed by the vaginal mucosa, while minimizing excretion from the vagina. Examples of thickening agents include: solid fats; propylene glycol; solid fat mixture from Heb. F. / NF / JPE, glyceryl ricinoleate, ethoxylated fatty alcohols (cetet-20, stearet-20) Heb. F. / NF (commercially available under the name OVUCIRE ^® 3460, GATIEFOSSÉ, Saint-Prix, France); solid fat mixture from Heb. F. / NF / JPE, glyceryl monooleate (type 40) from Heb. F. / NF (OVUCIRE WL 3264; mixture of solid fat from Hebrew F. / NF / JPE, glyceryl monooleate (type 40) from Hebrew F. / NF (OVUCIRE WL 2944); non-ionic surfactant being PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate; TEFOSE 63 or similar product; and a mixture of various solid fats (WITEPSOL ^® , Sasol Germany GmbH, Hamburg, Germany). Other thickening agents, for example, alginates, certain gums, such as xanthan gums, agar-agar, iota carrageenans, kappa carrageenans, etc. Some other compounds, such as in and polymers such as HPMC, PVC, and CMC. In accordance with various embodiments, the viscosity of pharmaceutical formulations can be from about 50 cp to about 1000 cp at 25 ° C. It will be easy for those skilled in the art to understand and choose from suitable thickening agents agents.

In accordance with embodiments, the thickening agent is a nonionic surfactant. For example, a polyethylene glycol ether or a diester of saturated or unsaturated fatty acids is a thickening agent in the form of a nonionic surfactant. In embodiments, the nonionic surfactant is a long chain (C16-C20) fatty acid polyethylene glycol ester and further is a long chain fatty acid ethylene glycol ether such as a PEG fatty acid ester or diesters of saturated or unsaturated C16-C18 fatty acids, for example, oleic , lauric, palmitic and stearic acids. In embodiments, the nonionic surfactant is a saturated long chain fatty acid polyethylene glycol ester and furthermore a saturated long chain fatty acid ethylene glycol ester, such as PEG and ethylene glycol esters of saturated C16-C18 fatty acids, for example palmitic and stearic acids. Such a nonionic surfactant may be PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate, of which TEFOSE 63 is a non-limiting example.

In accordance with embodiments, the nonionic surfactant used as a thickening agent is not hydrophilic and has good emulsifying properties. A representative example of such a surfactant is TEFOSE 63 with a hydrophilic-lipophilic balance (HLB) of about 9-10.

In accordance with embodiments, the pharmaceutical composition further comprises one or more mucoadhesive agents to improve the absorption of estradiol in the vagina. For example, a mucoadhesive agent may be present to facilitate adhesion of the pharmaceutical composition to the mucosa after its activation with water. In accordance with embodiments, polycarbophil is a mucoadhesive agent. In accordance with embodiments, mucoadhesive agents are unlimitedly represented, for example, polyethylene oxide polymers having a molecular weight of from about 100,000 to about 900,000; carbopol chitosans represented by acrylic acid polymers crosslinked with allyl succrose or allyl pentaerythrol; polymers of acrylic acid and C10-C30 alkyl acrylate crosslinked with allylpentaerythrol; a carbomer homopolymer or copolymer containing a block copolymer of polyethylene glycol and a long chain alkyl acid ester; etc. In accordance with embodiments, various hydrophilic polymers and hydrogels may be used as the mucoadhesive agent. In accordance with some embodiments, the polymers or hydrogels may swell upon contact with vaginal tissue or secretion, thereby enhancing the moisturizing and mucoadhesive effects. The choice and amount of the hydrophilic polymer may be based on the choice and amount of the dissolving agent. In some embodiments, the pharmaceutical composition comprises a hydrophilic polymer, but may optionally not contain a gelling agent. In embodiments containing hydrogel in an amount of from about 5% to about 10% by weight of the total composition, a hydrophilic polymer may be provided. In further embodiments, hydrogels may be used. The hydrogel can be represented by chitosan, which swells in the presence of water. In various embodiments, PEG-90M may be contained in the pharmaceutical composition of the cream. In some embodiments, the mucoadhesive agent is present in a pharmaceutical composition, in a soft gel capsule, or both.

In accordance with embodiments, the pharmaceutical compositions comprise one or more thermally reversible gels, typically with hydrophilic properties, unlimitedly represented, for example, by hydrophilic sucrose and other saccharide-based monomers (US Pat. No. 6,018,033, incorporated by reference).

In accordance with embodiments, the pharmaceutical composition further comprises a lubricant. In some embodiments, a lubricant may be present to facilitate preparation of the dosage form. For example, a lubricant may be added to prevent capsules or tablets from sticking together during production or storage. Any suitable lubricant may be used. For example, the lubricant is lecithin, which is a mixture of phospholipids.

In accordance with embodiments, the pharmaceutical composition further comprises an antioxidant. Any suitable antioxidant may be used. For example, butylated hydroxytoluene, butylated hydroxyanisole and TPGS vitamin E.

In accordance with embodiments, the pharmaceutical composition contains from about 20% to about 80% of a dissolving agent by weight, from about 0.1% to about 5% of a lubricant by weight, and from about 0.01% to about 0.1% of an antioxidant by weight .

The choice of excipient depends on factors such as, for example, the effect of the excipient on solubility and stability. Additional excipients used in various embodiments may be colorants and preservatives. Examples of dyes include FD&C dyes (e.g., blue dye No. 1 and red dye No. 40), D&C dyes (e.g. yellow dye No. 10), and opacifiers (e.g. titanium dioxide). In accordance with embodiments, colorants comprise from about 0.1% to about 2% of the pharmaceutical composition by weight. In accordance with embodiments, the preservatives in the pharmaceutical composition may be methyl paraben and propyl paraben in a ratio of about 10: 1 and in a proportion of about 0.005% and 0.05% by weight.

 In general, the dissolving agents, excipients, and other additives used in the pharmaceutical formulations described herein are non-toxic, pharmaceutically acceptable, and compatible with each other, and also maintain the stability of the pharmaceutical composition and the various components with respect to each other. In addition, a combination of the various components that make up the pharmaceutical compositions will help achieve the desired therapeutic effect when administered to a patient.

Solubility of estradiol

In accordance with embodiments, dissolving agents, which are mixtures of medium chain fatty acid glycerides, for example mono- and diglycerides or mono-, di- and triglycerides of C6-C12, C8-C12 or C8-C10 fatty acids, dissolve estradiol. As presented in the examples, good results were obtained using solvent agents, which are predominantly a mixture of C8-C10 mono- and diglycerides of saturated fatty acids or medium chain triglycerides (e.g., Miglyol 810 or 812). Longer chain glycerides are probably less suitable for dissolving estradiol.

The dissolving agent, which is propylene glycol monocaprylate (e.g. CAPRYOL) and 2- (2-ethoxyethoxy) ethanol (e.g. TRANSCUTOL), dissolves estradiol well.

PRODUCTION OF PHARMACEUTICAL COMPOSITION

In accordance with embodiments, a pharmaceutical composition is prepared by admixing estradiol with a pharmaceutically acceptable dissolving agent, unlimitedly represented, for example, by at least one medium chain fatty acid, such as medium chain fatty acids, which are at least one mono-, di- or triglyceride and derivatives or combinations thereof. In accordance with embodiments, the pharmaceutical composition may also contain at least one glycol or its derivatives and combinations or combinations of at least one glyceride and glycol. Glycol (s) can be used as a dissolving agent or for regulating viscosity, and therefore can be considered a thickening agent, which is further discussed herein. Optionally, other excipients are added, unlimitedly represented, for example, by antioxidants, lubricants, and the like. In accordance with embodiments, the pharmaceutical composition contains a sufficient amount of a dissolving agent to completely dissolve estradiol. However, it should be clearly understood that other volumes of the dissolving agent may be used depending on the desired solubility level of estradiol. It will be understood by those of ordinary skill in the art how to determine the volume of a dissolving agent and other excipients depending on the desired percentage of estradiol to be dissolved in the pharmaceutical composition.

In exemplary embodiments, GELUCIRE 44/14 (Lauroyl Macrogol-32 glycerides from Heb. F., NF Lauroyl Polyoxy-32 glycerides, US FDA IIG glycerides) are heated to about 65 ° C, and CAPMUL MCM is heated to about 40 ° C to facilitate the mixing of fat and non-ionic surfactant, although such heating is not required to dissolve estradiol.

The specific examples described herein provide additional principles and embodiments that illustrate the methods of manufacturing the pharmaceutical compositions described herein.

CARRIER

In general, the pharmaceutical compositions described herein are administered intravaginally using a carrier, for example, a capsule. In accordance with embodiments, the capsules are soft capsules made from materials well known to those skilled in the art, for example, gelatin. However, in accordance with embodiments, the carrier is integral with the pharmaceutical composition (i.e., the pharmaceutical composition is the carrier). In such embodiments, the pharmaceutical composition is a gel, cream, ointment, tablet, or other drug that is directly administered and absorbed into the vagina.

In accordance with embodiments, the pharmaceutical compositions described herein are contained in capsules, such as soft gelatine capsules. In accordance with embodiments, such capsules comprise one or more of the following agents: hydrophilic gel-forming bioadhesive (eg, mucoadhesive) agents; lipophilic agent; a gelling agent for a lipophilic agent or a hydrodispersing agent. In accordance with embodiments, the hydrophilic gelling bioadhesive agent is one of carboxyvinyl acid; hydroxypropyl cellulose; carboxymethyl cellulose; gelatin; xanthan gum; guar gum; aluminum silicate; or mixtures thereof. In accordance with embodiments, the lipophilic agent is one of a liquid triglyceride; solid triglyceride (with a melting point of about 35 ° C); carnauba wax; Coconut oil or mixtures thereof. In accordance with embodiments, the gelling agent is a hydrophobic colloidal silicon oxide. In accordance with embodiments, the hydrodispersing agent is one of: polyoxyethylene glycol; Polyoxyethylene glycol 7-glyceryl cocoate; or mixtures thereof.

In accordance with embodiments, the carrier is designed to facilitate administration of the drug. In accordance with embodiments, the carrier is sized to provide comfortable placement within the vagina. In accordance with embodiments, the carrier may have a different geometric shape. For example, the carrier may be in the form of a drop, a truncated cone, a cylinder, a cylinder with a larger “apical” part, or other forms suitable for insertion into the vagina and facilitating it. In accordance with embodiments, the carrier is used with the applicator. In accordance with other embodiments, the carrier is inserted by fingers.

In FIG. 2 shows a carrier 200, which is a pharmaceutical composition 202 and a capsule 204. A width of 208 indicates the thickness of the capsule 204, for example, about 0.274 cm (0.108 in). The distance from one end of the carrier 200 to the other is indicated by a distance of 206, for example, about 1.75 cm (0.690 inches). The size of the carrier 200 may also be described by an arc having a radius of a given length. For example, arc 210 bounded by the outer surface of gelatin 204 is an arc with a radius of about 0.480 cm (0.189 inches) in length. Arc 212, bounded by the inner surface of capsule 204, is an arc with a radius of about 0.238 cm (0.0938 inches) in length. Arc 214, bounded by the outer surface of gelatin 204 and opposite to arc 210, is an arc with a radius of about 0.274 cm (0.108 in) in length. Other capsules may be available in other sizes. In accordance with embodiments, capsule 204 has the same or similar dimensions with the above ratios relative to each other.

In accordance with embodiments, the carrier is designed to remain in the vagina until the pharmaceutical formulations are released. In accordance with embodiments, the carrier is dissolved in the vagina and absorbed by the vaginal tissue along with the pharmaceutical composition, which minimizes vaginal discharge. In such embodiments, the carrier is made of non-toxic components, for example, gelatin.

Design Parameters for Intravaginal Pharmaceutical Formulations

In accordance with embodiments, the pharmaceutical composition is designed to maximize preferred characteristics that promote patient adherence to treatment (patients who interrupt treatment before completing the prescribed course of treatment) without compromising efficacy. Preferred characteristics are presented, for example, by the absence or reduction of irritation in comparison with other hormone replacement pessaries, the absence or reduction of vaginal discharge of a pharmaceutical composition or carrier in comparison with other hormone replacement pessaries, the absence or reduction of the remainder of the pharmaceutical composition or carrier in the vagina, facilitation of administration compared to other hormone replacement pessaries or improved drug efficacy compared to AI with other similar pharmaceutical formulations.

In accordance with embodiments, the pharmaceutical composition does not cause irritation or minimizes it. Irritation in a patient is expressed by pain, prurit (itching), soreness, excessive discharge, swelling, or other similar symptoms. Irritation in the patient leads to low adherence to treatment. Pharmaceutical formulations that do not cause irritation or cause less irritation are defined in comparison with competing hormonal pessaries, including tablets, creams or other forms of intravaginal administration of estrogen.

According to embodiments, the pharmaceutical compositions do not have a systemic effect (for example, ingestion of estradiol in the bloodstream), which increases the safety of use. In accordance with other embodiments, the pharmaceutical formulations described herein have a significantly lower systemic effect (e.g., ingestion of estradiol into the bloodstream) compared to LPS.

In accordance with embodiments, the pharmaceutical composition does not leave a residue in the vagina. On the contrary, the pharmaceutical composition and the carrier are almost completely absorbed or dispersed, without leaving an unabsorbed residue and without causing unpleasant sensations from an unabsorbed or undispersed drug. The absence of residue is determined in comparison with other drugs for intravaginal administration or can be determined objectively when examining vaginal tissues. For example, some other drugs for intravaginal administration contain starch, which can cause more abundant vaginal discharge after drug administration. In some embodiments, the pharmaceutical formulations described herein provide a shorter amount, duration, and frequency of excretion after drug administration compared to other intravaginal drugs (eg, compressed tablets).

In accordance with embodiments, the pharmaceutical composition reduces the amount of vaginal discharge compared to other pessaries, including hormone-administered pessaries. Ideally, vaginal discharge is eliminated, minimized or reduced in comparison with competing drugs.

In accordance with embodiments, the pharmaceutical compositions described herein are administered by fingers. In accordance with embodiments, the pharmaceutical formulations are inserted with fingers approximately 5 cm (two inches) into the vagina without the use of an applicator. In accordance with embodiments, the pharmaceutical compositions are also formulated for administration by an applicator, if necessary. In accordance with some embodiments, due to the fact that the focus of the BBA is in the proximal part of the vagina (towards the vaginal opening), the pharmaceutical compositions described herein are designed to be inserted into the proximal part of the vagina.

During an extensive experimental program, various medium chain fatty acid glycerol and propylene glycol esters demonstrated one or more of the preferred characteristics for the development of a medicament for humans. In accordance with embodiments, the solvent agent has been selected from at least one of a solvent and a co-solvent. Suitable solvents and cosolvents are any mono-, di- or triglycerides and glycols, as well as combinations thereof.

In accordance with embodiments, the pharmaceutical composition is administered in the form of a gelatin capsule carrier. In accordance with these embodiments, the pharmaceutical composition is a liquid pharmaceutical composition. In accordance with embodiments, the carrier is a soft capsule, for example, a soft gelatin capsule. Thus, the pharmaceutical composition of such embodiments is encapsulated in a soft gelatin capsule or other soft capsule.

In accordance with embodiments, the pharmaceutical composition comprises estradiol, which is at least about 80% soluble in a dissolving agent, which is one or more C6-C14 mono-, di- and triglycerides of medium chain fatty acids and optionally a thickening agent. In accordance with embodiments, the pharmaceutical composition comprises estradiol, which is at least about 80% soluble in one or more of the mono-, di- and triglycerides of C6-C12 medium chain fatty acids, for example, in one or more of C6-C14 triglycerides, for example, in one or more of C6-C12 triglycerides, such as one or more of C8-C10 triglycerides. Such embodiments, in particular, imply that estradiol is soluble at least 80%. Such embodiments, in particular, imply that estradiol is soluble at least 90%. Such embodiments, in particular, imply that estradiol is soluble at least 95%. Such embodiments, in particular, imply that estradiol is completely soluble.

As noted above, liquid pharmaceutical formulations will be liquid at room temperature or at body temperature. For example, in some embodiments, the pharmaceutical composition described herein is a liquid composition that is contained within a soft gel capsule. Gels, solid fats or other solid dosage forms that will not be liquid at room temperature or at body temperature are less desirable in embodiments of a liquid pharmaceutical formulation.

The thickening agent serves to increase viscosity, for example, to about 10,000 cP (10,000 mPa-s), usually not more than about 5,000 cP, and most often from about 50 to 1,000 cP. In embodiments, a nonionic surfactant, such as GELUCIRE or TEFOSE, may be solid at room temperature and require melting to effectively mix with a dissolving agent. However, in these embodiments, the obtained pharmaceutical composition remains liquid, although with a higher viscosity, but not solid.

In accordance with embodiments, the pharmaceutical composition contains estradiol, a medium chain dissolving agent, and a thickening agent, which are administered as a soft carrier capsule. In addition, other ingredients, for example, colorants, antioxidants, preservatives, and the like, may be included. However, the addition of other ingredients should be in amounts that cannot substantially alter the solubility of estradiol, the pharmacokinetics of the pharmaceutical composition, or the effectiveness of the pharmaceutical composition. Other factors to consider when selecting ingredients for a pharmaceutical composition include irritation, vaginal discharge, vaginal discharge, and other significant factors, such as those that will lead to a decrease in patient adherence to treatment. Other ingredients that are implied include: fats or esters of fatty acids, lecithin, mucoadhesive agents, gelling agents, dispersing agents, and the like.

WAYS

In accordance with embodiments, the pharmaceutical compositions described herein can be used to treat BBA, including treating at least one of the symptoms of BBA, including but not limited to vaginal dryness, irritation or itching of the vagina or vulva, disorder urination, dyspareunia, and vaginal bleeding during intercourse. In accordance with embodiments, treatment methods generally relate to women.

In accordance with embodiments, the pharmaceutical compositions described herein can be used to treat urination disorders with estrogen deficiency. In accordance with embodiments, the pharmaceutical compositions described herein can be used to treat dyspareunia or vaginal bleeding during intercourse.

In accordance with embodiments, treatment of BBA, urination disorders with estrogen deficiency, dyspareunia, and vaginal bleeding during intercourse occurs by intravaginal administration of pharmaceutical formulations. According to embodiments in which the carrier is in the form of a capsule, the patient receives the capsule and introduces it into the vagina, where the capsule is decomposed, and the pharmaceutical composition is released into the vagina where it is absorbed by the vaginal tissue. In some embodiments, the pharmaceutical composition is completely absorbed by the vaginal tissue. In some embodiments, the pharmaceutical composition is almost completely absorbed by the vaginal tissue (for example, at least about 80% by weight, at least about 85% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, at least about 98% by weight, or at least about 99% by weight of the composition). In accordance with embodiments, the capsule is inserted with fingers approximately 5 cm (two inches) into the vagina, however, the depth of administration, in general, can be any that allows the absorption of almost the entire pharmaceutical composition. In accordance with embodiments, the capsule may also be administered using an applicator that places it in the vagina at the required depth, as described herein.

In accordance with embodiments in which the pharmaceutical composition is a cream, gel, ointment, or other similar preparations, the pharmaceutical composition is administered by fingers, as is well known in the art.

After the pharmaceutical composition is released into the vagina, estradiol is locally absorbed. For example, after administering a pessary to a patient in the proximal part of the vagina, a therapeutically effective concentration of estradiol is provided for 24 hours in the proximal part of the vagina.

In accordance with embodiments, the duration of administration of the pharmaceutical composition of the present disclosure may be carried out by any safe methods that have been prescribed by the attending physician. In accordance with embodiments, the patient must administer a pharmaceutical composition (eg, capsule) intravaginally every day for 14 days, and then twice a week.

In accordance with embodiments, pharmaceutical formulations are administered intravaginally with estrogen-based drugs (or progestin or progestin and estrogen-based drugs) for oral administration, patches, creams, gels, sprays, transdermal therapeutic systems or other parenteral estrogen-based drugs the introduction, each of which may contain natural, bioidentical, synthetic or other obtained estrogens or progestins. In accordance with embodiments, the regulation of estrogen concentrations in the blood stream, created by administering the pharmaceutical compositions described herein, if any, does not provide for the use of this composition as an additional drug for co-administration with another drug based on estrogen and regulation of estrogen concentration in blood flow associated with the introduction of the latter. In accordance with other variants of implementation of drugs based on estrogen for co-administration provide an additional effect, which will determine the attending physician of the patient.

In accordance with embodiments, the efficacy and safety of the pharmaceutical formulations described herein can be determined to treat the symptoms of BBA. In accordance with embodiments, the extent, effect, cytological diagnosis, histological diagnosis, and variability of the BBA can be described using various criteria for determining the efficacy and safety of pharmaceutical formulations described herein, or, if otherwise accepted in the art, now or later development. The criteria are based on published guidelines for the treatment of VVA with estradiol from the US Food and Drug Administration (FDA).

DETERMINATION OF EFFICIENCY

In accordance with embodiments, administration of the pharmaceutical formulations described herein has led to the treatment of BBA as well as the improvement of one or more symptoms associated with the BBA. In patients with BBA, the vaginal canal is narrowed both in length and in diameter, and in the vaginal canal the number of vaginal cells saturated with glycogen decreases, which provide hydration and elasticity. In addition, the vaginal wall may become thin, anemic, dry, or sometimes inflamed (atrophic vaginitis). Such changes can manifest themselves in the form of various symptoms, collectively referred to as BBA. Such symptoms are unlimitedly represented, for example, by an increase in the pH of the vagina; violation of the integrity of the vaginal epithelium, a decrease in vaginal secretion or a decrease in the density of the epithelium; prurite; vaginal dryness; dyspareunia (pain or bleeding during intercourse); urinary tract infections; or discoloration of the skin of the vagina. In accordance with the options for implementation, effectiveness is defined as an improvement in the condition of atrophy of the vulva and vagina in a patient to that which was in the premenopausal period. In accordance with the embodiments, the change is defined as the alleviation of the severity of one or more symptoms of atrophy, assessed at the inclusion stage (screening, 1 day) and compared with the assessment performed on day 15 (completion of treatment). The severity of the symptoms of atrophy can be assessed on a scale from 0 to 3, where, for example, absent = 0, mild = 1, moderate = 2 or severe = 3. A similar classification is used to assess the patient's condition before treatment; determine the correct course of treatment; for example, inter alia, the dosage, frequency and duration of administration of the drug; and evaluate the results after completion of treatment.

One of the symptoms of BBA is an elevated vaginal pH. In additional aspects of the present disclosure, treatment with the pharmaceutical formulations described herein has resulted in a decrease in vaginal pH. In accordance with embodiments, a decrease in vaginal pH is defined as a decrease in vaginal pH from the level at the inclusion stage (during screening) to the level of 15 days. In some embodiments, a pH of 5 or more may be associated with BBA. In some embodiments, the pH is measured using pH test strips that are applied to the vaginal wall. In some embodiments, a change in vaginal pH is a change in a patient's vaginal pH by less than about 5.0. In some embodiments, the patient's vaginal pH may be less than about 4.9; 4.8; 4.7; 4.6; 4,5; 4.4; 4.3; 4.2; 4.1; 4.0; 3.9; 3.8; 3.7; 3.6 or 3.5.

In accordance with embodiments, treatment with the pharmaceutical formulations described herein improves the vaginal maturation index. Maturation index is defined as a change in cell composition. In accordance with embodiments and with respect to BBA, a change in the composition of cells is defined as a change in the composition or quantity of parabasal, intermediate and superficial vaginal cells, expressed as a percentage, for example, as a change in the composition or number of parabasal cells of the vagina as compared to or relative to a change in composition or quantity superficial vaginal cells. A patient with symptoms of BBA often has an increase in the number of parabasal cells and a decrease in the number of surface cells (for example, less than about 5%) compared with women who do not suffer from BBA. Conversely, a patient with a decrease in the severity of symptoms, if she does not respond to the treatment differently, there may be an improvement in the value of the maturation index, in particular, a decrease in the number of parabasal cells or an increase in the number of surface cells compared with the indicators at the inclusion stage (during screening ) In embodiments, a decrease in the number of parabasal cells is defined as a decrease in the percentage of parabasal cells; the percentage reduction may be at least about 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15% or 10% of the number of parabasal cells. In embodiments, the percentage reduction may be at least about 54% of the number of parabasal cells. In embodiments, an increase in the number of surface cells is defined as an increase in the percentage of surface cells; the increase in the percentage of surface cells can be at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of the number of surface cells. In further embodiments, the percentage increase may be at least about 35% of the number of surface cells.

In some embodiments, an improvement in the maturation index value is defined as a change over time. For example, a change in the composition of cells determined at the stage of inclusion (during screening) at 1 day, compared with the composition of cells determined at 15 days. A change in the composition of cells can also be defined as a change in the number of parabasal cells over time, optionally in addition to determining changes in parabasal and surface cells, as described above. Such cells can be obtained from the epithelium of the vaginal mucosa during a standard gynecological examination and examined by taking a swab from the vagina.

In various further aspects of the present disclosure, treatment with the pharmaceutical formulations described herein has resulted in one of an increase in the number of surface cells; decrease in the number of parabasal cells; increase in the number of intermediate cells.

In further aspects of the present disclosure, blood samples may be taken to determine hormone levels, in particular estradiol levels. In some embodiments, blood samples can be taken from a patient and estradiol levels (pg / ml) can be measured. In some embodiments, estradiol levels can be measured after 0 hours (for example, during the administration of the first dose of the drug), after 1 hour (for example, after the first dose of the drug), after 3 hours and after 6 hours. In some embodiments, blood samples may be taken on day 8 (for example, after the first dose of the drug is administered) and on day 15 (for example, one day after the last dose of the drug is administered on day 14). In some embodiments, descriptive statistics of plasma estradiol concentrations for each sampling time and observed values of C _max and T _max can be determined, and the AUC value calculated.

In some embodiments, the pessary may contain about 25 μg of estradiol. In such cases, administering a pessary to a patient can provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) a corrected geometric mean maximum plasma concentration (C _max ) of estradiol ranging from about 19 pg * h / ml to about 29 pg * h / ml (e.g., from 19.55 pg * h / ml to about 28.75 pg * h / ml); 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estradiol, from about 75 pg * h / ml to about 112 pg * h / ml (for example, from 75.82 pg * h / ml to about 111.50 ) In some embodiments, administering a pessary to a patient may provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) an adjusted geometric mean maximum plasma concentration (C _max ) of estrone of about 9 pg * h / ml to about 14 pg * h / ml (e.g., from 9.17 pg * h / ml to about 13.49 pg * h / ml); 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone, from about 43 pg * h / ml to about 65 pg * h / ml (for example, from 43.56 pg * h / ml to about 64.06 pg * h / ml). In some embodiments, administering a pessary to a patient can provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) a corrected geometric mean maximum plasma concentration (C _max ) of estrone sulfate from about 416 pg * h / ml to about 613 pg * h / ml (e.g., from 416.53 pg * h / ml to about 612.55 pg * h / ml); 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, from about 3598 pg * h / ml to about 5291 pg * h / ml (for example, from 3598.04 pg * h / ml to about 5291, 24 pg * h / ml).

In some embodiments, the pessary may contain about 10 μg of estradiol. In such cases, administering a pessary to a patient can provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) a corrected geometric mean maximum plasma concentration (C _max ) of estradiol ranging from about 12 pg * h / ml to about 18 pg * h / ml (e.g., from 12.22 pg * h / ml to about 17.98 pg * h / ml); 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estradiol, from about 42 pg * h / ml to about 63 pg * h / ml (for example, from 42.18 pg * h / ml to about 62.02 pg * h / ml). and 3) a corrected geometric mean time to reach maximum plasma concentration (T _max ) of estradiol of about 1 hour to about 3 hours (for example, from 1.49 hours to about 2.19 hours). In some embodiments, administering a pessary to a patient may provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) an adjusted geometric mean maximum plasma concentration (C _max ) of estrone of about 4 pg * h / ml to about 7 pg * h / ml (e.g., from 4.38 pg * h / ml to about 6.44 pg * h / ml); 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone, from about 20 pg * h / ml to about 31 pg * h / ml (for example, from 20.60 pg * h / ml to about 30.30 pg * h / ml); and 3) a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone from about 4 hours to about 8 hours (for example, from 4.99 hours to about 7.34 hours). In some embodiments, administering a pessary to a patient may provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) a corrected geometric mean maximum plasma concentration (C _max ) of estrone sulfate from about 10 pg * h / ml to about 16 pg * h / ml (e.g., from 10.34 pg * h / ml to about 15.20 pg * h / ml); 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, from about 56 pg * h / ml to about 84 pg * h / ml (for example, from 56.61 pg * h / ml to about 83, 25 pg * h / ml); and 3) a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone sulfate of about 4 hours to about 7 hours (for example, from 4.67 hours to about 6.86 hours).

In some embodiments, the pessary may contain about 4 μg of estradiol. In such cases, administering a pessary to a patient can provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) a corrected geometric mean maximum plasma concentration (C _max ) of estradiol ranging from about 4 pg * h / ml to about 8 pg * h / ml; 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estradiol, from about 16 pg * h / ml to about 26 pg * h / ml; and 3) a corrected geometric mean time to reach maximum plasma concentration (T _max ) of estradiol of about 0.25 hours to about 2 hours. In some embodiments, administering a pessary to a patient may provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) an adjusted geometric mean maximum plasma concentration (C _max ) of estrone of about 1 pg * h / ml to about 3 pg * h / ml; 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone, from about 8 pg * h / ml to about 13 pg * h / ml; and 3) a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone from about 1 hour to about 4 hours. In some embodiments, administering a pessary to a patient can provide a number of parameters obtained from a patient’s blood plasma sample, including one or more of the following parameters: 1) a corrected geometric mean maximum plasma concentration (C _max ) of estrone sulfate from about 4 pg * h / ml to about 7 pg * h / ml; 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, comprising from about 22 pg * h / ml to about 34 pg * h / ml; 3) the adjusted geometric mean time to reach the maximum plasma concentration (T _max ) of estrone sulfate, from about 1 hour to about 3 hours.

The pharmaceutical composition described herein can provide almost completely topical application of estradiol. For example, plasma levels of estradiol, estrone and estrone sulfate measured in a patient’s plasma after administration of the pharmaceutical composition described herein may be statistically similar to levels measured after administration of a placebo composition (i.e., a similar composition without estradiol) . Accordingly, in some embodiments, the plasma levels of estradiol, estrone and estrone sulfate measured after administration of the pharmaceutical composition described herein may be low compared to LPS formulations.

In some embodiments, the pessary may contain from about 1 μg to about 25 μg of estradiol. After administration of the pessary to the patient, a blood plasma sample taken from him can give a corrected geometric mean of the maximum plasma concentration (C _max ) of estradiol, which is less than about 30 pg * h / ml. For example, administering a pessary to a patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estradiol, which is less than about 18 pg * h / ml. In some embodiments, administering the pessary to the patient provides a corrected geometric mean area under the 0-24 estradiol curve (AUC) of less than about 112 pg * h / ml. For example, administering a pessary to a patient provides an adjusted geometric mean area under the curve (AUC) of 0-24 estradiol, which is less than about 63 pg * h / ml.

In some embodiments, administering the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone, which is less than about 14 pg * h / ml. For example, administering a pessary to a patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone, which is less than about 7 pg * h / ml. In some embodiments, administering the pessary to the patient provides a corrected geometric mean area under the 0-24 estrone curve (AUC), which is less than about 65 pg * h / ml. For example, administering a pessary to a patient provides an adjusted geometric mean area under the AUC curve of 0-24 estrone, which is less than about 31 pg * h / ml.

In some embodiments, administering the pessary to the patient provides a corrected geometric mean maximum plasma concentration (C _max ) of estrone sulfate, which is less than about 613 pg * h / ml. For example, administering a pessary to a patient provides a corrected geometric mean of the maximum plasma concentration (C _max ) of estrone sulfate, which is less than about 16 pg * h / ml. In some embodiments, administering the pessary to the patient provides a corrected geometric mean area under the (AUC) curve of 0-24 estrone sulfate, which is less than about 5291 pg * h / ml. For example, administering a pessary to a patient provides an adjusted geometric mean area under the (AUC) curve of 0-24 estrone sulfate, which is less than about 84 pg * h / ml.

In further aspects of the present disclosure, capsule decomposition may be described. In some embodiments, the decomposition and absorption of the carrier (the presence or absence of the carrier after administration) can be described on day 1 of treatment (for example, 6 hours after the first dose of the drug) and on day 15 (for example, one day after the last dose of the drug 14 day).

Statistical Measurements

In accordance with embodiments, the pharmacokinetics of the pharmaceutical composition described herein is determined by statistical analysis. In accordance with embodiments, an analysis of variance ("ANOVA") or covariance analysis ("ANCOVA") is used to evaluate the difference between a patient who is being treated with a pharmaceutical composition containing an active pharmaceutical composition (eg, an estradiol-based pharmaceutical composition) and a patient who receives treatment with a placebo drug (for example, the same pharmaceutical composition, but without estradiol) or a comparison drug. An ordinary specialist in this field will understand how to conduct a statistical analysis of the selected data.

EXAMPLES

The following are examples of pharmaceutical formulations, carriers, and combinations thereof. Methods for their preparation are also described. In addition, the data obtained using the pharmaceutical compositions described herein are provided.

EXAMPLE 1 Pharmaceutical Composition

In embodiments, estradiol is prepared and combined with one or more pharmaceutically acceptable dissolving agents. Estradiol is prepared as an ingredient in the pharmaceutical category, often as micronized estradiol, however other forms may also be used. In embodiments, the pharmaceutical composition comprises estradiol in a dosage of from about 1 μg to about 50 μg. In embodiments, the pharmaceutical composition comprises 10 μg of estradiol. In embodiments, the pharmaceutical composition comprises 25 μg of estradiol.

In embodiments, for the formation of a pharmaceutical composition, estradiol is combined with one or more pharmaceutically acceptable dissolving agents and optionally with other excipients. In embodiments, the solvent agent is one of CAPMUL MCM, MIGLYOL 812, GELUCIRE 39/01, GELUCIRE 43/01, GELUCIRE 50/13 and TEFOSE 63.

Each of GELUCIRE 39/01 and GELUCIRE 43/01 has an HLB value of 1. GELUCIRE 50/13 has an HLB value of 13. TEFOSE 63 has an HLB value of 9 to 10.

Various combinations of pharmaceutically acceptable dissolving agents for estradiol were tested, as shown in Table 1.

Table 1
Capmul MCM ("MCM"), Gelucire 39/01 ("39/01"), Gelucire 43/01 ("43/01"), Gelucire 50/13 ("50/13") and Tefose ("Tefose 63" ) # Delivery system Attitude State of aggregation at room temperature Physical state at 37 ° C after ~ 30 minutes Viscosity (cP) Melting time at 37 ° C Dispersion in water at 37 ° C 1 MCM: 39/01 8: 2 Solid Clear liquid 50 at 37 ° C Start: 6 min End: 12 min Small drops of fat on the surface 2 MCM: 39/01 7: 3 Solid Clear liquid Start: 9 min End: 19 min 3 MCM: 39/01 6: 4 Solid Clear liquid Start: 20 min End: 32 min 4 MCM: 43/01 8: 2 Solid Particulate fluid 5 MCM: 43/01 7: 3 Solid Particulate fluid 6 MCM: 50/13 9: 1 Liquid/
muddy Liquid/
muddy 140 at 25 ° C Transparent after 20 min Homogeneous cloudy dispersion 7 MCM: 50/13 8: 2 Liquid/
muddy Liquid/
muddy 190 at 25 ° C Homogeneous cloudy dispersion 8 MCM: 50/13 7: 3 Semi-liquid Semi-liquid 9 MCM: TEFOSE 63 9: 1 Semi-liquid Liquid/
muddy 150 at 25 ° C Start: 1 min End: 5 min Homogeneous cloudy dispersion 10 MCM: TEFOSE 63 8: 2 Semi-liquid Semi-liquid 240 at 25 ° C Homogeneous cloudy dispersion eleven MCM: TEFOSE 63 7: 3 Semi-liquid Semi-liquid 380® 25 ° C Semi-solid after 30 min at 37 ° C, does not melt at 41 ° C Homogeneous cloudy dispersion 12 MIGLYOL 812: 50/13 9: 1 Semi-liquid Semi-liquid 140 at 25 ° C 2 phases, surface fat thirteen MIGLYOL 812: TEFOSE 63 9: 1 Liquid/
muddy Liquid/
muddy 90 at 25 ° C Start: 1 min End: 5 min 2 phases, surface fat

The pharmaceutical formulations of Table 1, which were liquid or semi-liquid at room temperature, were tested using a Brookfield viscometer (Brookfield Engineering Laboratories, Middleboro, Mass.) At room temperature. The pharmaceutical compositions shown in Table 1, which were solid at ambient temperature, were tested using a Brookfield viscometer at 37 ° C.

The pharmaceutical compositions shown in table 1, which were solid at room temperature, were aged at 37 ° C to determine their ability to melt. The viscosity of gels can be an important property when encapsulating the composition. For example, in some cases it is necessary to heat the composition before filling them with gelatin capsules. In addition, the ability of the composition to melt may be important after the introduction of the composition into the body. For example, in some embodiments, the composition will melt at temperatures below about 37 ° C. Pharmaceutical composition 11 (Capmul MCM / Tefose 63), for example, did not melt at 37 ° C or 41 ° C.

The dispersion was evaluated in the pharmaceutical formulations shown in Table 1. The dispersion was evaluated by adding 300 mg of each delivery system to 100 ml of water at 37 ° C without agitating and observing the mixing ability. The results ranged from the formation of droplets of fat on the surface to the separation of phases into homogeneous but turbid dispersions. In general, it is believed that formulations that can be easily dispersed in an aqueous solution will have better dispersibility after administration. However, it was unexpectedly discovered, as shown in examples 7-9, that formulations that were not readily dispersible in an aqueous solution (e.g., composition 13) and instead formed two phases after being introduced into an aqueous solution, proved to be most effective when introduced into the human body.

EXAMPLE 2 Media

In embodiments, the pharmaceutical composition is administered in the form of a gelatin capsule carrier. The gelatin carrier capsule is, for example, gelatin (e.g. gelatin, NF (150 Bloom, type B)), hydrolyzed collagen (e.g. GELITA ^® , GELITA AG, Eberbach, Germany), glycerin, special sorbitol or other excipients in proportions that are well known to specialists in this field. Special sorbitol can be purchased from suppliers and have a tendency to soften and moisturize the composition.

Various carriers have been developed, as shown in Table 2, these are A-F gels. In Table 2, each A-F carrier is characterized by a proportion of one or more components.

table 2
Carrier Gelatin Capsules A B C D E F Ingredient % weight./weight. % weight./weight. % weight./weight. % weight./weight. % weight./weight. % weight./weight. Gelatin, NF (150 Bloom, Type B) 41.0 41.0 41.0 41.0 43.0 43.0 Glycerin 99.7%, F. USA 6.0 6.0 6.0 6.0 18.0 18.0 Special Sorbitol, F. USA 15.0 15.0 15.0 15.0 GELITA® (hydrolyzed collagen) 3 3.0 Lemon acid 0.1 0.5 1 0.1 Purified water 35.0 37.9 37.5 37.0 36.0 38.9 Total 100.0 100.0 100.0 100.0 100.0 100.0 Dissolution of gel strips, on average 3 (500 ml of distilled H ₂ O, 50 rpm at 37 ° C) 48 min (42,45,58) 50 min (50.50.50) 75 min (76.75.74) 70 min (70.71.70) Dissolution of gel strips, an average of 3
(500 ml buffer with pH 4, 50 rpm at 37 ° C) 70 min 78 min 82 min

Each A-F gelatin carrier capsule was prepared at a temperature in the range of about 45 ° C to about 85 ° C. Each molten A-F carrier was cast in the form of a film, dried and cut into strips. The strips were cut into equal parts weighing about 0.5 g and with a thickness of about 0.5 mm. The strips were placed in a dissolution vessel of type 2 according to F. USA with water or buffer solution with pH 4, and the time required for complete dissolution was recorded (see table 2). Carrier A was most rapidly dissolved in water and a pH 4 buffer solution.

EXAMPLE 3 Pharmaceutical Formulations and Carriers

Various combinations of pharmaceutical formulations were obtained from Table 1 and Table 2. The combinations are shown in Table 3.

Table 3 Study Pharmaceutical composition Attitude Lot size
g Carrier 1 MCM: 39/01 8: 2 750 A 2 MCM: 50/13 8: 2 750 A 3 MCM: TEFOSE 63 8: 2 750 A 4 MCM: TEFOSE 63 8: 2 750 B 5 MIGLYOL 812: TEFOSE 63 9: 1 750 A

An aliquot of each pharmaceutical composition from table 3 is from about 300 mg to about 310 mg. The batch size is shown in Table 3. When encapsulating the delivery system, each about 300-310 mg of the pharmaceutical composition was encapsulated in about 200 mg of a gelatin carrier capsule. Thus, for example, in Study 1, the pharmaceutical composition designated MCM: 39/01 was encapsulated in a gelatin capsule carrier A, and the total encapsulated weight was from about 500 mg to about 510 mg. The size of the aliquot arbitrarily depends on the concentration of estradiol and the desired size of the gelatin carrier capsule. It will be easy for those skilled in the art to understand how to adjust the amount of estradiol in a pharmaceutical formulation to accommodate a given carrier size when the pharmaceutical formulation is encapsulated in a carrier.

EXAMPLE 4 Solubility of estradiol

In various studies, dissolving agents have been tested to determine their ability to dissolve 2 mg of estradiol in a pharmaceutical composition with a total weight of 100 mg. Solvents were considered suitable if the solubility of estradiol in the dissolving agent was higher or equal to about 20 mg / g. The initial solubility was determined by dissolving micronized estradiol in various dissolving agents until the estradiol was saturated (the dissolving agent with estradiol was balanced for 3 days), filtering off insoluble estradiol and analyzing the resulting pharmaceutical composition to determine the concentration of estradiol using HPLC.

Table 4
Solubility of Solvents (* indicates bibliographic source) Ingredient Solubility (mg / g) Peg 400 105 * Propylene glycol 75 * Polysorbate 80 36 * TRANSCUTOL HP 141 CAPMUL PG8 31,2

EXAMPLE 5 Pharmaceutical Formulations

The following pharmaceutical formulations are intended.

Gel mass

Ingredient % weight./weight. Amount / Lot (kg) Gelatin 150 Bloom, Bone, NF 41.00 82.00 Hydrolyzed Gelatin 3.00 6.00 Glycerin 99.7% 6.00 12.00 Special Sorbitol, NF 15.00 30.00 Dye Opatint White G-18006 1.20 2.40 Dye Opatine Red DG-15001 0.06 0.12 Purified Water, F. USA 33.74 67.48 Total 100.00

Pharmaceutical composition 1: 1 0 mcg estradiol

Ingredients Amount per capsule
(mg) % weight./weight. Number/
the consignment Estradiol hemihydrate, micronized, F. USA 0.010 0.003 CAPMUL® MCM, NF (glyceryl caprylate / caprate or medium chain mono- and diglycerides) 240.0 79,997 GELUCIRE® 50/13 (stearoylpolyoxyl-32 NF glycerides) 60.0 20,0 600.0 mg Total 300,0 100.0

Pharmaceutical composition 2:10 mcg estradiol

Ingredients Amount per capsule
(mg) % weight./weight. Number/
the consignment Estradiol hemihydrate, micronized, F. USA 0.010 0.003 MIGLOYL® 812 (medium chain triglyceride) 270,0 89,997 TEFOSE® 63 (a mixture of PEG-6 stearate, ethylene glycol palmitostearate or PEG-32 stearate; polyoxyl 6 and polyoxyl
32 palmitostearate / glycol stearate) 30,0 10.0 Total 300,0 100.0

Pharmaceutical composition 3: 25 mcg estradiol

Ingredients Amount per capsule
(mg) % weight./weight. Number/
the consignment Estradiol hemihydrate, micronized, F. USA 0.026 * 0.009 MIGLOYL® 812 (medium chain triglyceride) 270,0 89,991 TEFOSE® 63 (a mixture of PEG-6 stearate, ethylene glycol palmitostearate or PEG-32 stearate; polyoxyl 6 and polyoxyl
32 palmitostearate / glycol stearate) 30.02 10.0 Total 300,0 100.0 * 1.0 mg estradiol equivalent to 1.03 mg estradiol hemihydrate

Pharmaceutical composition 4: 4 mcg estradiol

Ingredients Amount per capsule
(mg) % weight./weight. Number/
the consignment Estradiol hemihydrate, micronized, F. USA 0.0041 * 0.001 MIGLOYL® 812 (medium chain triglyceride) 269.99 89,999 TEFOSE® 63 (a mixture of PEG-6 stearate, ethylene glycol palmitostearate or PEG-32 stearate; polyoxyl 6 and polyoxyl 32 palmitostearate / glycol stearate) 30,0 10.0 Total 300,0 100.0 * 1.0 mg estradiol equivalent to 1.03 mg estradiol hemihydrate

EXAMPLE 6 Process

In FIG. 1 shows an embodiment of a method for producing a pharmaceutical composition that contains estradiol dissolved in a CapmulMCM / Gelucire dissolving agent and encapsulated in a soft gelatin capsule carrier 100. In step 102, the CapmulMCM is heated to 40 ° C ± 5 ° C. Heating may be carried out using any suitable means. The heating may be carried out in any suitable vessel, such as a stainless steel vessel. Using the same general method, other pharmaceutical formulations can be obtained by replacing various excipients, including a dissolving agent.

In step 104, GELUCIRE is mixed with CapmulMCM to formulate a finished solvent. At 104, any form of GELUCIRE that has been used herein may be used. For example, in step 104, one or more of GELUCIRE 39/01, GELUCIRE 43/01, and GELUCIRE 50/13 may be used. Those of ordinary skill in the art will know that mixing is carried out, for example, using a paddle mixer, mixer, mixer, or other similar devices that are used to mix pharmaceutical formulations. Step 104 may be carried out in an atmosphere of an inert or relatively inert gas, such as nitrogen. Mixing can be carried out in any suitable vessel known to those of ordinary skill in the art, for example, in a stainless steel vessel or in a steel tank.

At step 106, estradiol is mixed with a dissolving agent. In embodiments, estradiol in micronized form is mixed with a dissolving agent. In other embodiments, estradiol is added in non-micronized form. Mixing can be facilitated using a paddle mixer, mixer, mixer or other similar devices that are used to mix pharmaceutical formulations. Step 106 may be carried out in an atmosphere of an inert or relatively inert gas, such as nitrogen.

However, in embodiments, the addition of estradiol can be carried out prior to step 104. In this regard, steps 104 and 106 are interchangeable in terms of time or can be carried out simultaneously with each other.

At 110, a gelatin carrier is made. At 110, any of the gelatinous carriers described herein can be used. In embodiments, gelatin, hydrolyzed collagen, glycerin and other excipients are mixed at a temperature in the range of from about 45 ° C to about 85 ° C and are made in the form of a film. Mixing can be carried out in a steel tank or other container used for the preparation of gelatin carriers. Mixing can be facilitated by using a paddle mixer, mixer, mixer or other devices that are used to mix the contents of gelatin carriers. Step 110 may be carried out in an atmosphere of an inert or relatively inert gas, such as nitrogen. In embodiments, the gelatin carrier mixture is degassed before being used to encapsulate the pharmaceutical composition.

At step 112, the pharmaceutical composition is encapsulated in a gelatin carrier in accordance with rules well known to those of ordinary skill in the art. In step 112, a soft gelatin carrier capsule is prepared by combining the pharmaceutical composition obtained in step 106 with a gelatin carrier in step 110. The gelatin can be wrapped around the material, encapsulating it in whole or in part, or it can also be injected into the gelatin or otherwise the pharmaceutical composition obtained in step 106 is introduced.

In embodiments, step 112 is carried out in a suitable matrix to give the product the desired shape. The resulting vaginal soft gel capsules can have a different geometric shape. For example, vaginal soft gel capsules may be in the form of a drop, a truncated cone, a cylinder, a cylinder with a larger “apical” part, as shown in FIG. 2, or other forms suitable for administration to the vagina. The finished pharmaceutical composition, encapsulated in a soft gelatin carrier, can be administered by fingers or using an applicator.

EXAMPLE 7 Study of the effectiveness of the pharmaceutical composition based on estradiol for the treatment of vulvovaginal atrophy (BBA)

The purpose of this study was designed to evaluate the efficacy and safety of a pharmaceutical formulation containing 10 μg of estradiol (i.e., pharmaceutical formulation 2) for treating moderate or severe symptoms of BBA associated with menopause after 14 days of treatment, and also to evaluate the magnitude of the effect and the variability of the endpoints of vulvovaginal atrophy. In addition, the systemic effects of estradiol from one to many doses of the pharmaceutical composition were investigated.

This study was a randomized, double-blind, placebo-controlled study of Phase 1 to evaluate the efficacy and safety of the pharmaceutical composition in alleviating moderate or severe symptoms of vaginal atrophy associated with the menopause, as well as to study the systemic effects of estradiol after the intravaginal administration of the pharmaceutical composition once a day within 14 days.

Postmenopausal women who met the criteria for inclusion in the study were randomly assigned to one of two treatment groups (with pharmaceutical composition or placebo). During the screening period, patients were asked to independently evaluate the symptoms of BBA, including vaginal dryness, irritation or itching of the vagina or vulva, urination disorders, vaginal pain during intercourse, and vaginal bleeding during intercourse. The patients who were eligible to participate in the study were women with at least one moderate or severe symptom of BBA, according to their own assessment, which they determined to be the most painful.

Clinical evaluations were performed at the following time points:

• Screening period (up to 28 days);

• Visit 1 - Randomization / Inclusion visit (1 day);

• Visit 2 - Interim visit (8 day); and

• Visit 3 - Completion of treatment (15 day).

Patients eligible to participate in the study were randomized at a 1: 1 ratio to obtain a pharmaceutical formulation containing 10 μg of estradiol or the corresponding placebo preparation in the form of a vaginal soft gel capsule, and they themselves administered the first dose of the study drug in a clinical setting under the supervision of research staff. A series of blood samples to track the level of estradiol concentration was taken 0, 1, 3, and 6.0 hours after the first dose was administered on day 1. The patients stayed in the hospital until the 6-hour blood sampling period ended, and then they re-visited the hospital for an additional one-time blood sample to measure estradiol levels on day 8 (before the morning dose) and on day 15. The patients were provided with a sufficient amount of the test drug until the next scheduled visit, they were also instructed to inject the prescribed test drug into the vagina once a day at approximately the same time (± 1 hour) in the morning. Each patient received a diary in which she had to write down the date and time of administration of the dose of the studied drug daily. Patients re-visited the clinical facility on day 8 for an intermediate visit and on day 15 to assess completion of treatment and examinations after the study. Evaluation of the state of decomposition of the capsule was made by the researcher on day 1 (6 hours after the dose of the drug) and on day 15.

The study included a screening period of up to 28 days before randomization and a treatment period of 14 days. The choice of dosage (10 μg estradiol) and treatment regimen (once a day for two weeks) was based on FDA data on the safety and efficacy of LPS.

Number of patients (planned and analyzed)

Up to 50 (25 in each treatment group) postmenopausal women aged 40 to 75 years with moderate or severe symptoms of BBA were randomized into groups. 50 patients were included in the study, 48 patients completed the study, and 48 patients were analyzed.

Diagnosis and main criteria for inclusion

Fifty women were included in the study. Women were included in the postmenopausal period, aged from 40 to 75 years, the average age was 62.3 years. The average weight (in kg) of patients was 71.2 kg with a weight range from 44.5 kg to 100 kg. The average height of the patients (in cm) was 162.6 cm with a growth range from 149.9 cm to 175.2 cm; the average body mass index (in kg / m 2) was 26.8 kg / m ² with a range of values from 19 kg / m ² to 33 kg / m ² . The criteria for inclusion in the study included: an independent assessment of at least one of the symptoms of BBA as moderate or severe, for example, vaginal dryness, dyspareunia, irritation, burning or itching of the vagina or vulva, urinary disorders, vaginal bleeding during intercourse, which the patient identified as the most painful for her; ≤5% of surface cells with vaginal smear cytology; vaginal pH>5.0; and the level of estradiol concentration is ≤50 pg / ml. The study included patients who were otherwise healthy, based on the results of the preliminary examination, clinical laboratory tests, gynecological examination and mammography.

10 mcg of estradiol or placebo, dose and route of administration of the drug

Patients were randomized into groups (in a 1: 1 ratio) for independent intravaginal administration once a day for 14 days of one of the following drugs:

• Formulation A: The pharmaceutical composition of Example 5 (pharmaceutical composition 2: 10 μg estradiol); or

• Drug B: Placebo in the form of a vaginal soft gel capsule containing a composition similar to that of drug A, except for 10 μg of estradiol.

The estradiol formulation was a light pink, drop-shaped soft gel capsule. Drug B had the same composition, appearance, and route of administration as drug A, but did not contain estradiol.

Treatment duration

The study included a screening period of up to 28 days before randomization and a treatment period of 14 days.

Criteria for evaluation

Performance Assessment Endpoints:

• Change in the maturation index (percentage of parabasal, superficial and intermediate vaginal cells), established on the basis of a smear from the vagina, for the period from the inclusion stage (screening) to 15 days. Information on this endpoint is given in Tables 6-8.

• Change in vaginal pH over the period from the inclusion phase (screening) to 15 days. Information on this endpoint is given in table 9.

• Change in the severity of the most painful symptoms over the period from the inclusion stage (randomization) to 15 days: (1) vaginal dryness; (2) irritation, burning, or itching of the vagina or vulva; (3) urination disorders; (4) dyspareunia; (5) vaginal bleeding during intercourse. Information on this endpoint is given in tables 13 and 15.

• Change in the researcher’s assessment of the vaginal mucosa over the period from the inclusion stage (randomization) to 15 days. Information on this endpoint is given in Tables 18-21.

Unless otherwise indicated, the endpoints for evaluating efficacy were defined as the change from the time of visit 1 - randomization / inclusion visit (1 day) to visit 3 - completion of treatment (15 days), except for vaginal bleeding, which was defined as success or failure of treatment.

Other endpoints include:

• The main indicators of the state of the body, weight, changes during physical examination, gynecological examination and examination of the mammary gland, as well as adverse events that were evaluated as the endpoints of the safety assessment.

• Estradiol concentration level for each blood sampling time.

• The maximum concentration of estradiol in 1 day and the time of sampling the blood when the maximum concentration was reached.

• Degree of decomposition of the carrier to determine the amount of residue in the vagina after treatment.

The results of the assessment of plasma levels of estradiol are shown in table 5.

Table 5
Safety study results: The following is descriptive statistics of Cmax and Tmax values for plasma estradiol concentrations at 1 day. 10 mcg estradiol Placebo C _max T _max C _max T _max N 24 24 26 26 Mean ± ST Deviation 30.7 ± 7.47 2.12 ± 1.73 27.5 ± 17.26 4.00 ± 2.68 Geometric mean 29.9 - 24.7 - Median value 29.8 1.00 22.1 6.00 Min max 19.7; 52.3 1.00; 6.00 15.1; 90.0 0.00; 6.00 The coefficient of variation, % 24.3% 81.3% 62.9% 67.1%

Other endpoints:

(f) Ripening Index Results

Vaginal cytology data was obtained on the basis of vaginal smears taken from the posterior wall of the vagina, in accordance with standard procedures for assessing vaginal cytology during the screening period and during visit 3 — completion of treatment (15 days). A change in the maturation index was defined as a change in the composition of cells measured during visit 1 — inclusion visit (1 day) compared with the composition of cells measured during visit 3 — completion of treatment (day 15). A change in the percentage of surface, parabasal and intermediate cells obtained from the epithelium of the vaginal mucosa from a vaginal smear was recorded. The results of these evaluations are given in tables 6, 7 and 8.

Table 6
The main results of the effectiveness analysis for changing the maturation index, established on the basis of a smear from the vagina, for the period from the stage of inclusion (screening) to 15 days (percentage of parabasal cells) Population Statistics Estradiol
10 mcg Placebo The difference between treatments 90% CI difference P-value: 10 mcg of estradiol versus placebo All patients with prescribed treatment N 24 24 - - - Middle limit -544 4.80 -49.6 (-60.4; -38.8) <0.0001 Mean ± ST Deviation -53.8 ± 39.7 -5.4 ± 22.3 - - - Median value -60.0 -5.0 - - - Min max -100.0; 0,0 -60.0; 60.0 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

Table 7
The main results of the effectiveness analysis for changing the maturation index, established on the basis of a smear from the vagina, for the period from the stage of inclusion (screening) to 15 days (percentage of surface cells) Population Statistics Estradiol
10 mcg Placebo The difference between treatments 90% CI difference P-value: 10 mcg of estradiol versus placebo All patients with prescribed treatment N 24 24 - - - Middle limit 35,2 8.75 26.5 (15.4; 37.6) 0,0002 Mean ± ST deviation 35.2 ± 26.4 8.8 ± 18.7 - - - Median value 40,0 0,0 - - - Min max 0,0; 80.0 0,0; 90.0 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo in ANOVA, where treatment is a fixed effect.
2 P-value for comparing treatment with ANOVA, where treatment is a fixed effect.

Table 8
The main results of the effectiveness analysis for changing the maturation index, established on the basis of a smear from the vagina, for the period from the inclusion stage (screening) to 15 days (percentage of intermediate cells) Population Statistics Estradiol
10 mcg Placebo The difference between treatments 90% CI difference P-value ² : 10 mcg of estradiol compared to placebo All patients with prescribed treatment N 24 24 - - - Middle limit 18.7 -3.54 22.3 (11.1; 33.5) 0.0017 Mean ± ST deviation 18.5 ± 42.7 -3.3 ± 21.6 - - - Median value 22.5 -5.0 - - - Min max -60.0; 100.0 -60.0; 20,0 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

PH Change Results

The vaginal pH was measured during screening and during visit 3 — completion of treatment (15 days). Measurement of the pH level was carried out by applying a test strip to determine the pH to the vaginal wall. Patients who were included in the study were required to have a vaginal pH above 5.0 at the time of screening. The pH values were recorded in the individual registration card of the subject of study. Patients were advised not to have sex and to refrain from douching 24 hours before the measurements. The results of these evaluations are shown in Table 9.

Table 9
The main results of the effectiveness analysis for changing the pH level of the vagina during the period from the inclusion stage (screening) to 15 days Population Statistics Estradiol
10 mcg Placebo The difference between treatments 90% CI
difference ¹ P-value ² : 10 mcg of estradiol compared to placebo All patients with prescribed treatment N 24 24 - - - Middle limit -0.974 0.339 -0.635 (-0.900; - 0.368) 0,0002 Mean ± ST deviation -0.917 ± 0.686 -0.396 ± 0.659 - - Median value -1.00 -0,500 - - - Min max -2.00; 0,500 -1.50; 0,500 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

Data on the most painful symptoms

Patients were asked to indicate a symptom that they would designate as "the most painful." During the screening, questionnaires were distributed to all patients for self-assessment of the symptoms of BBA: (1) vaginal dryness; (2) irritation, burning, or itching of the vagina or vulva; (3) urination disorders; (4) dyspareunia; (5) vaginal bleeding during intercourse. Each symptom, except for vaginal bleeding during intercourse, was rated on a scale of 0 to 3, where 0 = absent, 1 = mild, 2 = moderate and 3 = severe. Vaginal bleeding during intercourse was evaluated on a binary scale: H = no bleeding; D = there is bleeding. Patient responses were recorded. All randomized patients were also given questionnaires for self-assessment of symptoms of BBA during visit 1 - randomization / inclusion visit (1 day) and visit 3 - completion of treatment (15 day). Patients daily wrote down their own grades in a diary, their answers were collected on day 8 and day 15 (completion of treatment). The results of the assessments before the introduction of the first dose of the drug, obtained during visit 1, were considered for statistical analyzes as data at the inclusion stage. These estimates are given in tables 10 and 11.

Table 10
Characteristics of symptoms of vaginal atrophy at the inclusion stage (population with prescribed treatment) Symptom of VVA Statistics 10 mcg estradiol Placebo 10 μg estradiol in comp. with placebo P-value ¹ Vaginal dryness N subjects 24 24 - Mean 2,292 2,375 0.68231 Irritation / burning / itching of the vagina or vulva N subjects 24 24 - Mean 0.875 1,333 0,08721 Pain, burning or sharp pain when urinating N subjects 24 24 - Mean 0.583 0.625 0.87681 Vaginal pain during intercourse N subjects2 12 12 - Mean 2,083 2,333 0.54281 Vaginal bleeding during intercourse N subjects2 12 12 Percentage3 25.00 33.33 0.31463 ¹ P-value for comparing treatment with ANOVA / ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.
² N = number of patients having an active sexual life at the stage of inclusion.
³ Percentage of patients with bleeding determined using Fisher's exact test.

Table 11
Additional efficacy analysis results for changing the severity of symptoms of vaginal atrophy over the period from the inclusion stage (randomization) to 15 days Symptom Statistical Method ¹ Middle limit The difference between treatments 90% CI difference ² P-value: 10 mcg of estradiol versus placebo Estradiol
10 mcg Placebo Vaginal dryness ANCOVA 0.980 0.729 0.251 -0.706; 0.204) 0.3597 Irritation/
burning / itching of the vagina or vulva ANCOVA 0.694 0.514 0.180 -0.59; 0.189) 0.4159 Pain / burning /
acute pain (during urination) ANCOVA 0.391 0.359 0,032 -0.263; 0.200) 0.8185 Vaginal pain during intercourse Anova 0,800 0,500 0,300 -1.033; 0.433) 0.4872 ¹ ANOVA model contains a fixed effect for treatment. ANCOVA adds to the model the level at the stage of inclusion as an independent variable.
² Confidence interval for the difference between the mean limit for 10 μg of estradiol and placebo.

Changes in the severity of the most painful symptom from the level at the stage of inclusion were determined in accordance with the assessment of symptoms of BBA, generally described above. Tables 13 and 14 provide a comparison between pharmaceutical composition 1 and placebo, in general, for the most painful symptom and symptom of vaginal atrophy. It is important to note that these measurements showed an upward trend, albeit statistically insignificant, on day 15.

Table 13
The main results of the effectiveness analysis for changing the severity of the most painful symptoms of BBA for the period from the inclusion stage (randomization) to 15 days Population Statistics 10 mcg estradiol Placebo The difference between treatments 90% CI
difference ¹ P-value ² : 10 mcg of estradiol compared to placebo All patients with prescribed treatment N 24 24 - - - Middle limit -1.043 -1.042 -0.002 (-0.497; 0.493) 0,9951 Mean ± ST deviation -1.043 ± 0.928 -1.042 ± 1.08 - - - Median value -1.00 -1.00 - - - Min max -3.00; 0.00 -3.00; 0.00 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo in ANOVA, where treatment is a fixed effect.
2 P-value for comparing treatment with ANOVA, where treatment is a fixed effect.

Table 14
Additional efficacy analysis results for changing the severity of symptoms of vaginal atrophy over the period from the inclusion stage (randomization) to 15 days Symptom Statistical Method ¹ Middle limit The difference between treatments 90% CI
difference ² TX-12-004-HR
in comparison with
placebo P-value TX-12-004-HR Placebo Dryness ANCOVA -0.980 -0.729 -0,251 (-0.706; 0.204) 0.3597 Irritation ANCOVA -0.694 -0.514 -0.180 (-0.549; 0.189) 0.4159 Pain (during sex) Anova -0,800 -0,500 -0,300 (-1.033; 0.433) 0.4872 Pain / burning /
acute pain (during urination) ANCOVA -0.391 -0.359 -0.032 (-0.263; 0.200) 0.8185 1 ANOVA model contains a fixed effect for treatment. ANCOVA adds to the model the level at the stage of inclusion as an independent variable.
2 Confidence interval for the difference between the mean value limit for TX-12-004-HR and placebo.

Symptom

Regarding the data on the most painful symptoms presented in Tables 13 and 14, the period for which these data were selected is generally considered insufficient to draw significant conclusions. However, the trends observed in this study suggest that the data will show an improvement for most of the most painful symptoms if these data are collected over a longer period of time.

The absence or presence of any vaginal bleeding during intercourse was also defined as one of the most painful symptoms. Data on vaginal bleeding during intercourse are presented in table 15.

Table 15
The main results of the effectiveness analysis for changing vaginal bleeding during intercourse for the period from the inclusion stage (randomization) to 15 days Treatment N * Brief information on vaginal bleeding for the period from the inclusion stage (randomization) to 15 days Bleeding / no bleeding (success) 2 Bleeding/
bleeding (failure) No bleeding /
bleeding (failure) No bleeding /
no bleeding (BI) 10 mcg estradiol 10 2 (100%) 0 0 8 Placebo 10 1 (20%) 3 1 5 P-value: 10 μg of estradiol in comparison with
placebo ¹ 0.1429 - - - * N = The total number of patients in each treatment group who had an active sex life at the inclusion stage and on day 15, and also gave an answer during both visits.
BI = No change - not taken into account in statistical comparison.
¹ P-value for comparing treatment from Fisher's exact test.
2 Percentage based on the number of patients assigned to the Success group or the Failure group (N = 2 for the estradiol group 10 μg; N = 5 for the placebo group

Estradiol Concentration / Pharmacokinetic Data

In this study, the systematic effects of estradiol after intravaginal administration of 10 μg of estradiol once a day for 14 days were studied. Descriptive statistics of blood plasma levels of estradiol measured during each sampling and the observed values of C _max and T _max were recorded in Tables 16 and 17. No statistically significant difference in systemic concentration of 10 μg of estradiol was observed compared with placebo, which gives reason to believe that estradiol does not enter the bloodstream, where it will have a systemic effect. On the contrary, it remains localized in the tissues; the effect of estradiol is thus considered localized at the injection site (i.e., in the vagina). The lower detection limits of the assays used to determine the pharmacokinetic data could affect the accuracy of the determination of the presented pharmacokinetic values. Additional pharmacokinetic studies were carried out using more accurate analyzes in examples 8 and 9.

In order to monitor the level of estradiol concentration during the study, blood samples were taken 0, 1, 3, and 6 hours after a dose of the drug was administered on day 1; before the dose on day 8 and before the dose on day 15. Every effort was made to take samples at the appointed time. Procedures for the selection and processing of samples for measuring the level of estradiol concentration in the blood were carried out in accordance with the procedure approved by the sponsor of the study and the main researcher. The determination of all concentrations of estradiol in blood plasma at the stage of inclusion and after treatment was carried out using approved bioanalytical (UHEC with tandem mass spectrometry) methods. These data are given in tables 16 and 17.

Table 16
Descriptive statistics of estradiol concentrations (pg / ml) for each sampling time Treatment Sampling time 10 mcg estradiol After 0 hours After 1 hour After 3 hours In 6 hours Before a dose of 8 days Before a dose of 15 days N 24 24 24 24 24 22 Mean ± ST deviation 20.1 ± 5.74 28.7 ± 5.89 25.7 ± 5.71 23.4 ± 7.91 21.4 ± 9.28 23.4 ± 8.72 Median value 20,2 28.9 24.7 22.3 20.7 20.7 Min max 2.63; 38.3 18.8; 43.9 19.3; 47.5 3.31; 52.3 2.09; 52,2 17.9; 54.7 Placebo N 26 26 26 26 25 24 Mean ± ST Deviation 20.5 ± 4.29 21.0 ± 6.14 19.0 ± 5.92 26.9 ± 17.36 29.9 ± 22.51 28.1 ± 16.80 Median value 20.8 20.8 20.9 21.7 21.6 21.1 Min max 4.03; 29.1 3.19; 41.2 3.15; 26.9 15.1; 90.0 15.0; 116.2 14.7; 81.3

Table 17
Descriptive statistics for C _max and T _max values for estradiol at 1 day 10 mcg estradiol Placebo C _max T _max C _max T _max N 24 24 26 26 Mean ± ST deviation 30.7 ± 7.47 2.12 ± 1.73 27.5 ± 17.26 4.00 ± 2.68 Geometric mean 29.9 - 24.7 - Median value 29.8 1.00 22.1 6.00 Min max 19.7; 52.3 1.00; 6.00 15.1; 90.0 0.00; 6.00 The coefficient of variation, % 24.3% 81.3% 62.9% 67.1%

Assessment of vaginal mucosa data

Researchers evaluated the appearance of the vaginal mucosa at 1 day (before the dose of the drug) and at 15 days. The color, integrity of the vaginal epithelium, density of the vaginal epithelium and vaginal secretion were evaluated according to the following degrees of severity: none, mild, moderate and severe, using a scale from 0 to 3, where 0 = none, 1 = mild, 2 = moderate and 3 = heavy. The results of these evaluations of the researchers are given in tables 18, 19, 20 and 21.

Table 18
The main results of the effectiveness analysis for changing the assessment of researchers of the vaginal mucosa (assessment of vaginal color) for the period from the inclusion stage (randomization) to 15 days Population Statistics Estradiol
10 mcg Placebo The difference between treatments 90% CI difference ¹ P-value ² : 10 mcg of estradiol compared to placebo All patients with prescribed treatment N 24 24 - - - Middle limit -0.199 -0.009 -0.191 (-0.434; 0.052) 0.1945 Mean ± ST deviation -0.333 ± 0.565 0.125 ± 0.741 Median value 0.00 0.00 - - - Min max -2.00; 0.00 -1.00; 2.00 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

Table 19
The main results of the effectiveness analysis for changing the assessment of researchers of the vaginal mucosa (assessment of the integrity of the vaginal epithelium) for the period from the inclusion stage (randomization)
up to 15 days Population Statistics 10 mcg estradiol Placebo The difference between treatments 90% CI
difference 1 P-value2: 10 mcg of estradiol versus placebo All patients with prescribed treatment N 24 24 - - - Middle limit -0.342 0.176 -0.518 (-0.726; - 0.311) 0.0001 Mean ± ST deviation -0.417 ± 0.584 0.250 ± 0.442 Median value 0.00 0.00 - - - Min max -1.00; 1.00 0.00; 1.00 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

Table 20
The main results of the effectiveness analysis for changing the assessment of researchers of the vaginal mucosa (assessment of the density of the vaginal epithelium) for the period from the inclusion stage (randomization)
up to 15 days Population Statistics Estradiol
10 mcg Placebo The difference between treatments 90% CI
difference ¹ P-value ² : 10 mcg of estradiol compared to placebo All patients with prescribed treatment N 24 24 - - - Middle limit -0.034 -0.133 0,099 (-0.024; 0.221) 0.1820 Mean ± ST deviation -0.125 ± 0.338 -0.042 ± 0.550 - - - Median value 0.00 0.00 - - - Min max -1.00; 0.00 -1.00; 1.00 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

Table 21
The main results of the effectiveness analysis for changing the assessment of researchers of the vaginal mucosa (assessment of vaginal secretion) for the period from the inclusion stage (randomization) to 15 days Population Statistics 10 mcg estradiol Placebo The difference between treatments 90% CI
difference ¹ P-value ² : 10 mcg of estradiol compared to placebo All patients with prescribed treatment N 24 24 - - - Middle limit -0.643 -0.274 -0.369 (-0.661; - 0.076) 0,0401 Mean ± ST deviation -0.792 ± 0.779 -0.125 ± 0.741 - - - Median value -1.00 0.00 - - - Min max -2.00; 1.00 -2.00; 2.00 - - - 1 Confidence interval for the difference between 10 μg of estradiol and placebo with ANCOVA, where treatment is a fixed effect and the level at the inclusion stage is an independent variable.
2 P-value for comparing treatment with ANCOVA, where treatment is a fixed effect, and the level at the inclusion stage is an independent variable.

Media Decomposition Data

Assessment of the degree of decomposition of the capsule in the vagina (presence or absence) at 1 day (6 hours after the dose of the drug) and on day 15. The results of this assessment are shown in Table 22.

Table 22
The state of decomposition of the capsule into the vagina on day 1 and day 15 10 mcg estradiol Placebo 1 day 15 day 1 day 15 day There are no signs of a capsule 23 (95.8%) 24 (100.0%) 26 (100.0%) 24 (92.3%) Signs of capsule present 0 (0.0%) 0 (0.0%) 0 (0.0%) 0 (0.0%) No assessment 1 (4.2%) 0 (0.0%) 0 (0.0%) 22 (7.7%)

Serum hormone concentration data were selected to measure serum estradiol levels. These data were used for inclusion during screening and were determined using standard methods of clinical chemistry.

Accuracy of measurements

The selection of performance criteria used in this study was based on FDA guidelines for research on estrogen and estrogen / progestin medications for the treatment of moderate or severe vasomotor symptoms associated with menopause and moderate or severe symptoms of vulvar and vaginal atrophy associated with Menopause (Food and Drug Administration, Guidance for Industry, Estrogen and Estrogen / Progestin Drug Products to Treat Vasomotor Symptoms and Vulvar and Vaginal Atrophy Symptoms - Recommendations for Clinical Evaluation. January 2003, incorporated herein by links).

The study used standard clinical, laboratory, and statistical procedures. All clinical laboratory procedures were generally accepted and met quality standards.

Statistical methods:

Efficiency:

Analysis of variance (ANOVA) was used to assess changes in level at the stage of inclusion of differences between patients who took 10 μg of estradiol and who took capsules with placebo at all endpoints of the evaluation of effectiveness, except for vaginal bleeding, to assess the magnitude and variability of the effect. In some cases, for example, for some symptoms of vaginal atrophy, the change from the level at the stage of inclusion (response after a dose) was compared with the value at the stage of inclusion (p <0.05), thus, the level at the stage of inclusion was introduced as an independent variable, which should be adjusted for this relationship (Covariance analysis, ANCOVA) Confidence intervals of 90% of the difference between the average endpoints for estradiol 10 μg and placebo were determined to assess the magnitude of the effect. The change in vaginal bleeding during intercourse from the level at the inclusion stage was evaluated in terms of the proportion of patients whose treatment ended in success or failure. Each patient who had bleeding at the inclusion stage and who did not report bleeding on day 15 was considered a patient whose treatment was successful. Each patient who reported bleeding on day 15 was considered a patient whose treatment failed, regardless of whether she reported bleeding at the inclusion stage or not. Patients whose bleeding was not observed either at the initial stage or on day 15 were classified as “unchanged” and were excluded from the statistical evaluation. The difference in the proportion of patients whose treatment was successful in the two groups was statistically evaluated using Fisher's exact test. The results of this difference in proportion are shown in Table 10.

Determination of adherence to treatment

Patients needed to keep a diary to mark adherence to treatment. Diaries were reviewed for adherence to treatment during the visit on day 8 and day 15. The total number of patients who demonstrated 100% adherence to the treatment regimen was 45 people (21 patients in the group with estradiol 10 μg and 24 patients in the group with placebo).

Due to the exploratory nature of the study, no adjustments were made regarding the set of endpoints.

Security:

The frequency and severity of all adverse events were descriptively summarized for each treatment group.

Results: Data from all forty-eight (48) patients who completed the study were included in the main efficacy analyzes. The results of the effectiveness analyzes are presented in tables 5, 6 and 7.

conclusions

Efficiency

Treatment with 10 μg of a pharmaceutical composition lasting two weeks resulted in a statistically significant higher average decrease in the percentage of parabasal cells than treatment with placebo (54% versus 5%, p <0.0001), which is shown in Table 6. At that at the same time, a significantly higher average value of an increase in the percentage of surface cells was observed in the group with the pharmaceutical composition (35%) than in the group with capsules with placebo (9%), while the difference was of high statistical significance (p = 0.0002), which shown in table 7. The difference in the decrease in pH between the group with the pharmaceutical composition (0.97 units) and the group with placebo (0.34 units) was only slightly more than 0.5 units, but this difference was noted as statistically significant (p = 0.0002), as shown in table 9.

While the decrease in the severity of the most painful symptoms was almost the same (~ 1 unit) for both groups with the pharmaceutical composition and placebo, the decrease in the severity of individual symptoms, such as vaginal dryness, irritation and pain during intercourse, was slightly better in the group with active treatment than in the placebo group. None of the differences between the two treatments, which amounted to ≤ 0.3 units, was considered statistically significant. There was no difference between the two types of treatment in terms of reducing pain / burning / acute pain during urination (the decrease was ~ 0.4 units). The duration of the study was not long enough to show a discrepancy in the most painful symptoms between the group with the pharmaceutical composition and the group with placebo. However, trends in the most painful symptoms suggest that significant differences can be observed between the two treatments over an appropriate period of time.

Treatment with capsules with 10 μg of estradiol for a duration of two weeks showed no statistically significant difference in reducing the severity of symptoms from the level at the inclusion stage in accordance with the researchers' assessment of the color or density of the vaginal epithelium. Compared to placebo, capsules with a pharmaceutical composition nevertheless showed a higher statistically significant decrease in the severity of atrophic effects on the integrity of the vaginal epithelium (-0.34 versus 0.18, p = 0.0001) and on vaginal secretion (-0.64 versus - 0.27, p = 0.0401).

Descriptive statistical analyzes (mean, median, geometric mean, standard deviation, coefficient of variation, minimum and maximum, C _max and T _max ) were carried out at estradiol concentrations during each sampling, at a maximum concentration of 1 day and at the maximum concentration. The results of this assessment are given in tables 16 and 17.

The pharmaceutical composition containing 10 μg of estradiol exceeded the placebo in terms of its characteristics in terms of improving the maturation index, lowering the vaginal pH, reducing the atrophic effect on the integrity of the epithelium and vaginal secretion. The lack of statistical significance between the two treatments for reducing the severity of the most painful symptoms and individual symptoms of vaginal atrophy, such as dryness, irritation, pain during intercourse and pain / burning / acute pain during urination, is not unexpected, given the small number of subjects and short duration of treatment. Only a few patients experienced vaginal bleeding during intercourse, which does not allow any meaningful assessment of this symptom of vaginal atrophy.

Of the 48 patients included in the study, 45 showed 100% adherence to the treatment regimen. Of the remaining three patients, one stopped participating in the study for personal reasons, and the other two did not administer one dose of the drug due to an adverse event.

Security

Despite the fact that on day 1 the average value of the maximum concentration of estradiol in the blood plasma for the pharmaceutical composition was slightly higher than this value for placebo (geometric mean = 1.21: study drug (10 μg estradiol) by 21%> placebo ), no statistically significant difference was noted. However, the analysis methods were controversial, which casts doubt on the pharmacokinetic data. Additional pharmacokinetic studies were conducted in examples 8 and 9.

No significant adverse events occurred during the study.

In general, a pharmaceutical formulation containing 10 μg of estradiol was well tolerated with intravaginal administration once a day for 14 days.

EXAMPLE 8: pharmacokinetic study (composition 25 μg)

A pharmacokinetic study was conducted to compare the 25 μg formulation (pharmaceutical formulation 3) described herein with LPS. The results of a pharmacokinetic study of estradiol are summarized in table 23. The P-values for these data demonstrate statistical significance, as shown in table 24.

Table 23
Statistical summary of comparative bioavailability data for studies of the average unscaled bioequivalence of estradiol, the limit of geometric mean values of estradiol, the ratio of average values and confidence interval of 90%, the study of bioequivalence in patients on an empty stomach / after eating (Study No.: ESTR-1K-500-12); 25 mcg estradiol dose Parameter Test drug N LPS N Attitude
(%) 90% CI C _max (pg / ml) 23.0839 36 42.7024 36 54.06 44.18-66.14 AUC _0-24 (pg.h / ml) 89,2093 36 292.0606 36 30.54 23.72-39.34

Table 24
P-values for table 23 the effect P value Cmax AUC _0-24 Treatment <0001 <0001 Sequence 0.4478 0.5124 Period 0.4104 0.7221

As described in Table 23, baseline-adjusted pharmacokinetic data show that the formulations described herein unexpectedly show a decrease in C _{max of} 54% and a decrease in AUC of 31% with respect to LPS. Such a result is desirable since estradiol is intended for local absorption only. These data indicate a decrease in circulating estradiol levels compared to LPS. Moreover, it should be noted that the levels of C _max and AUC of estradiol with respect to placebo are not statistically differentiable, which suggests that the compositions described herein have an insignificant systemic effect. As shown in table 24, there was no significant difference between the study drug and the comparison drug due to the effects of sequence and period. However, a significant difference was observed due to the treatment effect for both C _max and AUC.

The pharmacokinetics for the total circulating level of estrone, an estradiol metabolite, are given in table 25. These data show that the total circulating level of estrone for the formulations described herein caused a decrease in C _{max of} 55% for the circulating level of estrone and a decrease in AUC of 70% for circulating estrone levels.

Table 25
Statistical summary of comparative bioavailability data for studies of the average unscaled bioequivalence of estrone, the limit of geometric mean values, the ratio of average values and confidence intervals of 90%, the study of bioequivalence in patients on an empty stomach / after eating (Study No.: ESTR-1K-500-12); 25 mcg estradiol dose Parameter Test drug N LPS N Attitude
(%) 90% CI C _max
(pg / ml) 10.7928 36 23,5794 36 45.77 32.95 to 63.59 AUC _0-24
(pg.h / ml) 51,2491 36 165.4664 36 30.97 19.8-48.45

Table 26
P-values for table 25 the effect P value C _max AUC _0-24 Treatment 0,0002 <0.0001 Sequence 0.1524 0.0464 Period 0,0719 0.0118

There was a significant difference between the study drug and the comparison drug due to the treatment effect, in turn, there was no significant difference due to the effects of the sequence and period for C _max . For the AUC value between the study drug and the reference drug, a significant difference was observed due to the effects of treatment, sequence and period.

The pharmacokinetics for the total circulating estrone sulfate level are given in table 27. These data show that the total circulating estrone sulfate level for the pharmaceutical formulations described herein resulted in a 33% decrease in C _max and a 42% decrease in AUC for the circulating estrone sulfate .

Table 27
Statistical summary of comparative bioavailability data for studies of the average unscaled bioequivalence of estrone sulfate, the limit of geometric mean values of estrone sulfate, the ratio of average values and confidence intervals of 90%, a study of bioequivalence in patients on an empty stomach / after eating (Study no: ESTR-1K-500-12); 25 mcg estradiol dose Parameter Test drug N LPS N Attitude
(%) 90%
DI C _max
(pg / ml) 490.0449 36 730.5605 36 67.08 53.84-83.57 AUC _0-24
(pg.h / ml) 4232,9914 36 7323.0827 36 57.80 43.23-77.29

Table 28
P-values for table 27 the effect P value C _max AUC _0-24 Treatment 0.0042 0.0031 Sequence 0.5035 0.9091 Period 0.1879 0.3304

There was a significant difference between the study drug and the comparison drug due to the treatment effect, in turn, there was no significant difference due to the sequence and period effects for C _max and AUC.

EXAMPLE 9: pharmacokinetic study (composition 10 μg)

A pharmacokinetic study was conducted to compare the 10 μg formulation (pharmaceutical formulation 2) described herein with LPS. The results of a pharmacokinetic study of estradiol are set forth in Tables 29-40 and in FIG. 9-14.

A pharmacokinetic study was conducted to compare the pharmaceutical composition described herein containing 10 μg of estradiol with LPS. The results of a pharmacokinetic study of estradiol are set forth in Tables 29-34, which demonstrate that the pharmaceutical formulations described herein more effectively prevent systemic absorption of estradiol. Table 35 shows that the pharmaceutical formulations described herein showed a result 28% better than LPS for total blood concentrations of C _max and 72% better than LPS for AUC.

Table 29
Brief information about the pharmacokinetic parameters of the studied drug (I) estradiol - adjusted for the initial value (N = 34) Pharmacokinetic parameter Arithmetic mean ± standard deviation The coefficient of variation Median value Minimum Maximum C _max (pg / ml) 15.7176 ± 7.9179 50,3761 13,9000 6,5000 49,6000 AUC _0-24
(pg.h / ml) 53.0100 ± 19.5629 36.9041 49.9750 24,3000 95,1500 t _max (h) 1.98 ± 1.29 65.34 2.00 1.00 8.05

Table 30
Brief information about the pharmacokinetic parameters of the comparison drug (C) estradiol - adjusted for the initial value (N = 34) Pharmacokinetic parameter Arithmetic mean ± standard deviation The coefficient of variation Median value Minimum Maximum C _max (pg / ml) 24.1882 ± 11.9218 49.2877 24,1500 1,0000 55.3000 AUC _0-24 (pg.h / ml) 163.8586 ± 72.0913 43,9960 158.0375 2,0000 304.8500 t _max (h) 10.53 ± 5.58 52.94 8.06 2.00 24.00

Table 31
The geometric mean of the test drug (I) and the comparison drug (C) estradiol - adjusted for the initial value (N = 34) Pharmacokinetic parameter Geometric mean The studied drug (I) Comparison drug (C) C _max (pg / ml) 14,3774 20.3837 AUC _0-24 (pg.h / ml) 49,6231 132,9218 t _max (h) 1.75 9.28

Table 32
Statistical results of the study drug (I) in relation to the comparison drug (C) for estradiol - adjusted for the initial value (N = 34) Pharmacokinetic parameter Geometric limit Intraindividual coefficient of variation,% I / C ratio
% 90%
Confidence interval The studied drug (I) Comparison drug (C) Cmax (pg / ml) 14,4490 20.1980 60.68 71.54 * 56.82-90.08 AUC _0-24
(pg.h / ml) 49.7310 131,0400 70.64 37.95 * 29.21-49.31 * Comparison was defined as statistically significant using ANOVA (a = 0.05)

Pharmacokinetic data for the total concentration of estrone similarly demonstrated a degree of systemic exposure lower than that of LPS. Table 33 shows that for the pharmaceutical formulations described herein, systemic exposure was 25% less for Cmax and 49% for AUC.

Table 33
Brief information about the pharmacokinetic parameters of the studied drug (I) estrone - adjusted for the initial value (N = 33) Pharmacokinetic parameter Arithmetic mean ± standard deviation The coefficient of variation Median value Minimum Maximum C _max (pg / ml) 6.8485 ± 6.5824 96.1149 5,4000 1,3000 36,3000 AUC _0-24
(pg.h / ml) 34.7051 ± 27.9541 80,5476 30.8500 3.3500 116.7500 t _max (h) 9.12 ± 8.83 96.80 4.00 1.00 24.00

Table 34
Brief information about the pharmacokinetic parameters of the drug comparison (C) of estrone - adjusted for the initial value (N = 33) Pharmacokinetic parameter Arithmetic mean ± standard deviation The coefficient of variation Median value Minimum Maximum C _max (pg / ml) 8.8333 ± 7.1469 80.9086 6,7000 2,7000 30,3000 AUC _0-24
(pg.h / ml) 63.0042 ± 46.5484 73.8814 51,2800 8,8000 214,0000 t _max (h) 11.16 ± 7.24 64.95 10.00 4.00 24.00

Table 35
The geometric mean of the studied drug (I) and the comparison drug (C) of estrone - adjusted for the initial value (N = 33) Pharmacokinetic parameter Geometric mean The studied drug (I) Comparison drug (C) C _max (pg / ml) 5,1507 6,9773 AUC _0-24 (pg.h / ml) 24.2426 48.2377 t _max (h) 5.87 9.07

Table 36
Statistical results of the studied drug (I) in relation to the comparison drug (C) for estrone - adjusted for the initial value (N = 33) Pharmacokinetic parameter Geometric limit Intraindividual coefficient of variation,% I / C ratio
% 90%
Confidence interval The studied drug (I) Comparison drug (C) C _max (pg / ml) 5.1620 6.9280 47.59 74.50 * 61.69-89.97 AUC _0-24
(pg.h / ml) 24.1960 47.9020 73.66 50.51 * 38.37-66.50 * Comparison was defined as statistically significant using ANOVA (α = 0.05)

Pharmacokinetic data for the total concentration of estrone sulfate similarly demonstrated a lower systemic exposure than LPS. Table 37 shows that for the pharmaceutical formulations described herein, systemic exposure was 25% less for C _max and 42% for AUC.

Table 37
Brief information about the pharmacokinetic parameters of the test drug (I) estrone sulfate - adjusted for the initial value (N = 24) Pharmacokinetic parameter Arithmetic mean ± standard deviation The coefficient of variation Median value Minimum Maximum C _max (ng / ml) 13.9042 ± 7.0402 50,6339 11,1500 1,3000 39,0000 AUC _0-24
(ng.h / ml) 97.9953 ± 80.8861 82,5408 76.2750 5,1025 338,0000 t _max (h) 6.33 ± 4.56 71.93 4.00 4.00 24.00

Table 38
Brief information on the pharmacokinetic parameters of the drug of comparison (C) of estrone sulfate - adjusted for the initial value (N = 24) Pharmacokinetic parameter Arithmetic mean ± standard deviation The coefficient of variation Median value Minimum Maximum C _max (ng / ml) 19.2542 ± 11.3633 59,0173 15,2000 7,0000 53,7000 AUC _0-24
(ng.h / ml) 177.6208 ±
166,2408 93.5931 124,0000 20,0000 683,0500 t _max (h) 10.33 ± 5.58 54.05 10.00 2.00 24.00

Table 39
The geometric mean of the studied drug (I) and the comparison drug (C) of estrone sulfate - adjusted for the initial value (N = 24) Pharmacokinetic parameter Geometric mean The studied drug (I) Comparison drug (C) C _max (ng / ml) 12.1579 16.8587 AUC _0-24 (ng.h / ml) 66,5996 121,5597 t _max (h) 5.49 8.83

Table 40
Statistical results of the studied drug (I) in relation to the comparison drug (C) for estrone sulfate - adjusted for the initial value (N = 24) Pharmacokinetic parameter Geometric limit Intraindividual coefficient of variation,% I / C ratio
% 90%
Confidence interval The studied drug (I) Comparison drug (C) C _max (ng / ml) 12.3350 16.5470 48.02 74.55 * 59.43-93.51 AUC _0-24
(ng.h / ml) 68,5260 118.4170 73.87 57.87 * 41.68-80.35 * Comparison was defined as statistically significant using ANOVA (a = 0.05)

While pharmaceutical formulations and methods have been described in terms of embodiments that are currently considered practical and preferred, it should be understood that the disclosure should not be limited to the described embodiments. It is intended to encompass various modifications and similar arrangements contemplated by the spirit and scope of the claims, the scope of which should be accorded a broader interpretation so as to accommodate all such modifications and similar embodiments. The present disclosure covers, without exception, embodiments of the following claims.

Claims (97)

1. A pessary for use in the treatment of vulvovaginal atrophy, including: a liquid pharmaceutical composition comprising estradiol and / or estradiol hemihydrate, where the total concentration of estradiol and estradiol hemihydrate in said pessary is equivalent to a concentration of estradiol in the range of 1-25 μg, where 1 μg of estradiol is equivalent to 1.03 μg of estradiol hemihydrate; and a medium chain fat dissolving agent; where the liquid pharmaceutical composition is encapsulated in a capsule; and where the treatment implies a decrease in the pH of the vagina in the patient and / or implies a change in the composition of the cells in the patient, where a change in the composition of the cells implies a decrease in the number of parabasal cells of the vagina or an increase in the number of surface cells of the vagina. 2. The pessary of claim 1, wherein the medium chain fat contains at least one of a C6-C12 fatty acid or glycol or a monoglyceride, diglyceride or triglyceride ester thereof. 3. The pessary according to any one of paragraphs. 1 or 2, wherein the dissolving agent is at least one ester selected from the group consisting of caproic fatty acid ester, caprylic fatty acid ester, capric fatty acid ester, and combinations thereof. 4. The pessary according to any one of paragraphs. 1 or 2, in which the dissolving agent is a caprylic / capric acid triglyceride. 5. The pessary of claim 1, wherein the capsule is a soft gelatin capsule. 6. The pessary according to claim 1, including: a) caprylic / capric acid triglyceride; b) a nonionic surfactant comprising PEG-6 stearate and ethylene glycol palmitostearate. 7. The pessary according to any one of paragraphs. 1, 2 or 6, where the administration of a suppository to a patient in the proximal part of the vagina provides a therapeutically effective concentration of estradiol for 24 hours in the proximal part of the vagina. 8. The pessary according to any one of paragraphs. 1, 2 or 6, where the treatment involves a decrease in the pH of the vagina in the patient. 9. The pessary according to claim 8, where the treatment involves reducing the pH of the vagina of the patient to a value of less than 5.0. 10. The pessary according to any one of paragraphs. 1, 2 or 6, where treatment involves changing the composition of cells in a patient. 11. The pessary according to claim 10, where the change in the composition of the cells implies a decrease in the number of parabasal cells of the vagina. 12. The pessary according to claim 10, where the change in the composition of the cells implies an increase in the number of superficial cells of the vagina. 13. The pessary according to claim 12, where the number of parabasal cells of the vagina of the patient is reduced by at least 35% or 50%. 14. The pessary of claim 12, wherein the number of superficial vaginal cells increases by at least 5% or 35%. 15. The pessary according to any one of paragraphs. 1, 2 or 6, including: 0.010 mg estradiol hemihydrate; 270.0 mg of caprylic / capric acid triglyceride or MIGLOYL® 812; and 30 mg of a mixture of PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate or TEFOSE® 63. 16. The pessary according to any one of paragraphs. 1, 2 or 6, including: 0.026 mg estradiol hemihydrate; 270.0 mg of caprylic / capric acid triglyceride or MIGLOYL® 812; and 30.02 mg of a mixture of PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate or TEFOSE® 63. 17. The pessary according to any one of paragraphs. 1, 2 or 6, including: 0.0041 mg estradiol hemihydrate; 269.99 mg of caprylic / capric triglyceride MIGLOYL® 812; and 30 mg of a mixture of PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate or TEFOSE® 63. 18. The pessary according to any one of paragraphs. 1, 2 or 6, including: 0.0097 mg of estradiol; 270.0 mg of caprylic / capric acid triglyceride or MIGLOYL® 812; and 30 mg of a mixture of PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate or TEFOSE® 63. 19. The pessary according to any one of paragraphs. 1, 2 or 6, including: 0.025 mg estradiol; 270.0 mg of caprylic / capric acid triglyceride or MIGLOYL® 812; and 30.02 mg of a mixture of PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate or TEFOSE® 63. 20. The pessary according to any one of paragraphs. 1, 2 or 6, including: 0.0039 mg of estradiol; 269.99 mg of caprylic / capric triglyceride or MIGLOYL® 812; and 30 mg of a mixture of PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate or TEFOSE® 63. 21. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estradiol, ranging from 19 to 29 PG * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of 0-24 estradiol, from 75 to 112 pg * h / ml. 22. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estrone, ranging from 9 to 14 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone, ranging from 43 to 65 pg * h / ml. 23. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estrone sulfate, from 416 to 613 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, from 3598 to 5291 pg * h / ml. 24. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 10 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estradiol, ranging from 12 to 18 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estradiol, comprising from 42 to 63 pg * h / ml. 25. The pessary of claim 24, wherein the pessary additionally provides a corrected geometric mean time to reach maximum plasma concentration (T _max ) of estradiol of 1 to 3 hours. 26. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 10 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estrone, from 4 to 7 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of 0-24 estrone, ranging from 20 to 31 PG * h / ml. 27. The pessary according to claim 26, characterized in that the pessary additionally provides a corrected geometric mean time to reach maximum plasma concentration (T _max ) of estrone from 4 to 8 hours. 28. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 10 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estrone sulfate, from 10 to 16 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of 0-24 estrone sulfate, comprising from 56 to 84 pg * h / ml. 29. The pessary according to claim 28, wherein the pessary additionally provides an adjusted geometric mean time to reach the maximum plasma concentration (T _max ) of estrone sulfate ranging from 4 to 7 hours. 30. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 4 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estradiol, from 4 to 8 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estradiol, from 16 to 26 pg * h / ml. 31. The pessary according to claim 30, wherein the pessary additionally provides a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estradiol ranging from 0.25 to 2 hours. 32. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 4 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estrone, ranging from 1 to 3 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone, from 8 to 13 pg * h / ml. 33. The pessary of claim 32, wherein the pessary additionally provides a corrected geometric mean time to reach maximum plasma concentration (T _max ) of estrone ranging from 1 to 4 hours. 34. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 4 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the introduction of the pessary to the patient provides in a patient's blood plasma sample: 1) the adjusted geometric mean maximum plasma concentration (C _max ) of estrone sulfate, from 4 to 7 pg * h / ml; and 2) the adjusted geometric mean area under the curve (AUC) of _0-24 estrone sulfate, from 22 to 34 pg * h / ml. 35. The pessary according to claim 34, wherein the pessary additionally provides a corrected geometric mean time to reach the maximum plasma concentration (T _max ) of estrone sulfate ranging from 1 to 3 hours. 36. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the administration of the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estradiol, which is less than 30 pg * h / ml. 37. The pessary of claim 36, wherein administering the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estradiol that is less than 18 pg * h / ml. 38. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where administering the pessary to the patient provides a corrected geometric mean area under the o-24 estradiol curve (AUC) of less than 112 pg * h / ml. 39. The pessary of claim 38, wherein administering the pessary to the patient provides an adjusted geometric mean area under the (AUC) o-24 estradiol curve that is less than 63 pg * h / ml. 40. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the administration of the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone, which is less than 14 pg * h / ml. 41. The pessary of claim 40, wherein administering the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone, which is less than 7 pg * h / ml. 42. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where administering the pessary to the patient provides an adjusted geometric mean area under the o-24 estrone curve (AUC) of less than about 65 pg * h / ml. 43. The pessary of claim 42, wherein administering the pessary to the patient provides an adjusted geometric mean area under the (AUC) curve of o-24 estrone, which is less than 31 pg * h / ml. 44. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 μg to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the administration of the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone sulfate, which is less than 613 pg * h / ml. 45. The pessary of claim 44, wherein administering the pessary to the patient provides an adjusted geometric mean maximum plasma concentration (C _max ) of estrone sulfate, which is less than 16 pg * h / ml. 46. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 μg to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where administering the pessary to the patient provides an adjusted geometric mean area under the curve (AUC) of o-24 estrone sulfate, which is less than 5291 pg * h / ml. 47. The pessary of claim 46, wherein administering the pessary to the patient provides an adjusted geometric mean area under the (AUC) curve of o-24 estrone sulfate that is less than 84 pg * h / ml. 48. A pessary for use in the treatment of vulvovaginal atrophy, comprising a liquid pharmaceutical composition comprising 1 μg to 25 μg of estradiol, wherein the liquid pharmaceutical composition is encapsulated in a capsule; and where the administration of a suppository to a patient in the proximal part of the vagina provides a therapeutically effective concentration of estradiol for 24 hours in the proximal part of the vagina. 49. A method of treating vulvovaginal atrophy, the method involves administering to a patient in need of treatment a pessary according to any one of claims. 21-48. 50. The method according to p. 49, in which the treatment involves alleviating the severity of one or more symptoms taken from the following group of symptoms: vaginal dryness, pain during intercourse, irritation of the vagina or vulva, burning of the vagina or vulva, itching of the vagina or vulva, disorder urination and vaginal bleeding during intercourse. 51. The method according to p. 49, in which the treatment involves reducing the pH of the vagina in a patient. 52. The method according to p. 50, in which the treatment involves reducing the pH of the vagina in a patient.

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