KR20180100567A - Vaginal insertion type estradiol pharmaceutical composition and method - Google Patents

Abstract

Above all, soft gelatinous pharmaceutical compositions and formulations containing solubilized estradiol for the treatment of vulvar-vaginal atrophy (VVA) and feminine dysfunction (FSD) are disclosed herein.

Description

Vaginal insertion type estradiol pharmaceutical composition and method

Cross-reference to related application

This application claims the benefit of U.S. Provisional Application No. 62 / 264,309, filed December 7, 2015; U.S. Provisional Application No. 62 / 296,552, filed February 17, 2016; U.S. Provisional Application No. 62 / 324,838, filed April 19, 2016; 62 / 329,940, filed April 29, 2016; And 62 / 348,820, filed June 10, 2016, the disclosure of which is incorporated herein by reference in its entirety.

Technical field

The present application relates to pharmaceutical compositions, methods and devices related to hormone replacement therapy.

Postmenopausal women are often referred to as vaginal dryness, vaginal odor, vaginal or vulvar irritation or itching, dysuria (pain during urination, tingling or tingling), dyspareunia (vaginal pain associated with sexual activity) (&Quot; vulvo-vaginal atrophy " or " VVA ") with atrophic vaginitis or vocal fold and vaginal atrophy with symptoms including bleeding. Other symptoms are soreness; Frequency and urgency; Also includes discomfort and incontinence (" estrogen deficient urinary state (s) ") that occurs. One symptom of vaginal atrophy is an increase in vaginal pH, which creates an environment more susceptible to infection. The mucosal epithelium of patients with VVA has also been reported to show signs of severe atrophy and cytologic examination is accompanied by an increase in the number of epidermal cells and a decrease in the number of epidermal cells.

Each of these VVA related conditions exhibits symptoms associated with decreased estrogenation of the vulvar vaginal tissue and may even occur in women treated by oral administration of an estrogenic pharmaceutical product. VVA is most common in postmenopausal women, but it can occur at any time in a woman's life cycle. VVA symptoms also interfere with sexual activity and satisfaction. Women with female sexual dysfunction (FSD) will have nearly four times more VVA than women without FSD.

Estrogen therapy has proven to be very successful in modulating menopausal symptoms, including VVA and FSD. Several studies have shown that symptoms associated with vaginal atrophy are usually alleviated by systemic or topically applied estrogen therapy. Existing therapies have a number of problems, for example, complications from patients who have not completed or continued treatment due to problems associated with the type of treatment.

Thus, there remains a need in the art for treatment for VVA and FSD that overcomes this limitation.

Above all, new soft gelatin pharmaceutical compositions and formulations containing solubilized estradiol for the treatment of VVA are disclosed herein. The soft gelatinous pharmaceutical compositions are designed to alleviate the common limitations found by other vaginal forms of estradiol. Soft gel-like pharmaceutical compositions provide a more effective formulation with ease of vaginal administration, improved insertion safety, minimized vaginal discharge after administration, and improved efficacy, safety and patient compliance.

According to various aspects and embodiments of the disclosure, there is provided a soft gelatinous pharmaceutical composition as a treatment for postmenopausal women suffering from a moderate to severe symptom of VVA.

a) a therapeutically effective amount of estradiol; And b) a solubilizing agent comprising an intermediate chain oil is provided herein.

In some embodiments, the suppository comprises about 1 [mu] g to about 25 [mu] g of estradiol. For example, suppositories may contain from about 1 [mu] g to about 10 [mu] g of estradiol; And about 10 [mu] g to about 25 [mu] g of estradiol.

In some embodiments, estradiol is solubilized.

In some embodiments, the intermediate chain oil comprises at least one C6-C12 fatty acid or glycol, monoglyceride, diglyceride or triglyceride ester thereof.

In some embodiments, the solubilizer comprises at least one ester selected from the group consisting of esters of capro fatty acids, esters of caprylic fatty acids, esters of capric fatty acids, and combinations thereof. For example, the solubilizing agent may comprise caprylic / capry triglyceride.

In some embodiments, the suppository further comprises a capsule. For example, the capsule may be a soft gelatin capsule.

a) a therapeutically effective amount of estradiol; b) capryl / capric triglyceride; c) a nonionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And d) suppositories, including soft gelatine capsules, are also provided herein.

In some embodiments, the suppositories provided herein comprise about 25 μg of estradiol and the administration of the suppository to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 29 pg * hr / Calibrated geometric mean peak plasma concentration of estradiol (C _max ); And 2) a corrected curve geometric mean area (AUC) _0-24 of estradiol from about 75 pg * hr / ml to about 112 pg * hr / ml.

In some embodiments, the suppositories provided herein comprise about 25 μg of estradiol, and administration of the suppository to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 14 pg * hr / Calibrated geometric mean peak plasma concentration (C _max ) of < RTI ID = 0.0 > And 2) a corrected curve geometric mean area (AUC) _0-24 of the estrone from about 43 pg * hr / ml to about 65 pg * hr / ml.

In some embodiments, the suppositories provided herein comprise about 25 μg of estradiol and the administration of the suppository to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 613 pg * hr / Calibrated geometric mean peak plasma concentration (C _max ) of estrone sulfate in ml; And 2) a corrected curve geometric mean area (AUC) _0-24 of the estrone sulfate from about 3598 pg * hr / ml to about 5291 pg * hr / ml.

In some embodiments, the suppositories provided herein comprise about 10 [mu] g of estradiol, and administration of the suppositories to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 18 pg * hr / Calibrated geometric mean peak plasma concentration of estradiol (C _max ); And 2) a corrected curve geometric mean area (AUC) _0-24 of estradiol from about 42 pg * hr / ml to about 63 pg * hr / ml. In some embodiments, suppositories further provide a corrected geometric mean time (T _max ) for the peak plasma concentration of estradiol from about 1 hour to about 3 hours.

In some embodiments, the suppositories provided herein comprise about 10 [mu] g of estradiol and the administration of the suppository to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 7 pg * hr / Calibrated geometric mean peak plasma concentration (C _max ) of < RTI ID = 0.0 > And 2) a corrected curve geometric mean area (AUC) _0-24 of the estrone from about 20 pg * hr / ml to about 31 pg * hr / ml. In some embodiments, suppositories further provide a corrected geometric mean time ( _Tmax ) for peak plasma concentrations of estrone from about 4 hours to about 8 hours.

In some embodiments, the suppositories provided herein comprise about 10 [mu] g of estradiol and the administration of the suppositories to the patient comprises: 1) administering a therapeutically effective amount of at least about 10 pg * hr / ml to about 16 pg * hr / Calibrated geometric mean peak plasma concentration (C _max ) of estrone sulfate in ml; And 2) a corrected curve geometric mean area (AUC) _0-24 of estrone sulfate from about 56 pg * hr / ml to about 84 pg * hr / ml. In some embodiments, suppositories further provide a corrected geometric mean time (T _max ) for peak plasma concentrations of estrone sulfate from about 4 hours to about 7 hours.

In some embodiments, the suppositories provided herein comprise about 4 μg of estradiol and the administration of the suppository to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 8 pg * hr / Calibrated geometric mean peak plasma concentration of estradiol (C _max ); And 2) a corrected curve geometric mean area (AUC) of _0-24 of estradiol from about 16 pg * hr / ml to about 26 pg * hr / ml. In some embodiments, the suppository further provides a corrected geometric mean time (T _max ) for the peak plasma concentration of estradiol from about 0.25 hours to about 2 hours.

In some embodiments, the suppositories provided herein comprise about 4 μg of estradiol and the administration of the suppositories to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 3 pg * hr / Calibrated geometric mean peak plasma concentration (C _max ) of < RTI ID = 0.0 > And 2) a corrected curve geometric mean area (AUC) of _0-24 of the estrone from about 8 pg * hr / ml to about 13 pg * hr / ml. In some embodiments, suppositories further provide a corrected geometric mean time (T _max ) for the peak plasma concentration of estrone from about 1 hour to about 4 hours.

In some embodiments, the suppositories provided herein comprise about 4 μg of estradiol and the administration of the suppository to the patient comprises: 1) administering to the patient about 1 pg * hr / ml to about 7 pg * hr / Calibrated geometric mean peak plasma concentration (C _max ) of estrone sulfate in ml; And 2) a corrected curve geometric mean area (AUC) _0-24 of about 22 pg * hr / ml to about 34 pg * hr / ml of estrone sulfate. In some embodiments, suppositories further provide a corrected geometric mean time (T _max ) for peak plasma concentrations of estrone sulfate from about 1 hour to about 3 hours.

Suppositories comprising about 1 [mu] g to about 25 [mu] g of estradiol are also provided herein, and administration of suppositories to the patient comprises adjusting the corrected geometric mean peak plasma concentration ( _Cmax ) of estradiol to less than about 30 pg * hr / Lt; / RTI > For example, administration of a suppository to a patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of estradiol of less than about 18 pg * hr / ml.

In some embodiments, a suppository is provided comprising about 1 g to about 25 g of estradiol, and the administration of the suppository to the patient comprises a corrected curved geometric mean area (AUC) of estradiol of less than about 112 pg * hr / _0-24 . For example, administration of a suppository to a patient provides a corrected curve geometric mean area (AUC) _0-24 of estradiol of less than about 63 pg * hr / ml.

In some embodiments, suppositories are provided comprising about 1 g to about 25 g of estradiol, and administration of the suppositories to the patient provides a corrected geometric mean peak plasma concentration (C _max ) of the estrone of less than about 14 pg * hr / Lt; / RTI > For example, administration of a suppository to a patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of the estrone of less than about 7 pg * hr / ml.

In some embodiments, about 1㎍ to about 25㎍ are provided suppositories containing estradiol, the administration of the suppository of the patient is from about 65pg * hr / ㎖ less than the corrected geometric mean area under the curve (AUC) of estrone _{0 -24} . For example, administration of a suppository to a patient provides a corrected curve geometric mean area (AUC) _0-24 of estrone of less than about 31 pg * hr / ml.

In some embodiments, about 1㎍ to about 25㎍ are provided suppositories containing estradiol, the administration of the suppository of the patient is from about 613pg * hr / ㎖ less than estrone sulfate, the corrected geometric mean peak plasma concentration (C _max of ). For example, administration of a suppository to a patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of estrone sulfate less than about 16 pg * hr / ml.

In some embodiments, suppositories comprising about 1 [mu] g to about 25 [mu] g of estradiol are provided and administration of the suppositories to the patient comprises a corrected curved geometric mean area (AUC) of the estrone sulfate of less than about 5291 pg * hr / _0-24 . For example, administration of a suppository to a patient provides a corrected curve geometric mean area (AUC) _0-24 of estrone sulfate less than about 84 pg * hr / ml.

Suppositories comprising about 1 [mu] g to about 25 [mu] g of estradiol are further provided herein and administration of the suppositories to the proximal region of the patient's vagina provides a therapeutically effective concentration of estradiol over 24 hours in the proximal region of the vagina to provide.

The present disclosure also provides a method of treating an estrogen deficiency condition, the method comprising administering a suppository as provided herein to a patient in need thereof. In some embodiments, there is provided a method of treating abnormal tone-vaginal atrophy, the method comprising administering a suppository as provided herein to a patient in need thereof.

In some embodiments of the methods provided herein, the treatment is selected from the group consisting of vaginal dryness, dyspareunia, vaginal or vulvar irritation, vaginal or vulvar tingling, vaginal or vulvar itching, dysuria, and vaginal bleeding associated with sexual activity And reducing the severity of the symptoms.

In some embodiments of the methods provided herein, the treatment comprises reducing the vaginal pH of the patient. For example, treatment includes reducing the patient ' s vaginal pH to a pH of less than about 5.0.

In some embodiments of the methods provided herein, the treatment comprises a change in the cellular composition of the patient. For example, changes in cell composition include a decrease in the number of vaginal vaginal cells or an increase in the number of vaginal vaginal cells. In some embodiments, the number of vaginal vaginal cells in a subject is reduced by at least about 35% (e.g., at least about 50%). In some embodiments, the number of epidermal vaginal cells increases by at least about 5% (e.g., at least about 35%).

A method of reducing vaginal discharge after administration of a suppository is further provided herein, the method comprising administering to a patient in need thereof a suppository as provided herein, wherein the vaginal discharge after administration of the suppository comprises the administration of a reference drug Compared with vaginal discharge after administration.

Methods of treating female sexual function in a female subject requiring treatment of female sexual function are also provided herein. The method comprises administering to the subject a vaginal suppository as described herein. In some embodiments, the method comprises: (a) administering a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A pharmaceutical composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) administering to the subject a vaginal suppository comprising soft gelatin capsules; The vaginal suppository comprises from about 1 microgram to about 25 microgram of estradiol; Estradiol is the only active hormone in the vaginal suppository. In some embodiments, the vaginal suppository does not contain a hydrophilic gel-forming bioadhesive agent in the solubilizer. In some embodiments, treating feminine sexual function involves increasing sexual desire, excitement, lubrication, satisfaction, and / or orgasm of the subject.

The above-mentioned features and objects of the present disclosure will become more apparent with reference to the following description taken in conjunction with the accompanying drawings, in which like reference numerals identify like elements,
1 is a flow diagram illustrating a process in accordance with various embodiments of the present invention.
Figure 2 illustrates a suppository in accordance with various embodiments of the present invention.
Figure 3 is a linear plot of mean plasma estradiol-time adjusted concentration versus time (n = 34).
FIG. 4 is a half-log plot of the adjusted mean concentration of plasma estradiol versus baseline (n = 34) over time.
Figure 5 is a linear plot of mean plasma estrone-baseline adjusted concentration over time (n = 33).
Figure 6 is a semi-log plot of the mean plasma estrone-baseline adjusted concentration over time (n = 33).
Figure 7 is a linear plot of mean plasma estrone sulfate-baseline adjusted concentration over time (n = 24).
Figure 8 is a semi-log plot of the mean plasma estrone sulfate-baseline adjusted concentration over time (n = 24).
Figure 9 is a schematic diagram of the study.
Figure 10 shows the percentage change in epidermal cells at 12 weeks compared to placebo.
Figure 11 shows the percentage change in epidermal cells at 2 weeks, 6 weeks, 8 weeks and 12 weeks compared to placebo.
Figure 12 shows the percentage change in epidermal cells per dose for 2 weeks, 6 weeks, 8 weeks and 12 weeks compared to placebo, respectively.
Figure 13 shows the percentage change in basal cell count at 12 weeks compared to placebo.
Figure 14 shows the percentage change in basal cell numbers at 2 weeks, 6 weeks, 8 weeks and 12 weeks compared to placebo.
Figure 15 shows the percentage change in basal cells per dose for 2 weeks, 6 weeks, 8 weeks, and 12 weeks compared to placebo, respectively.
Figure 16 shows the percentage change in pH at 12 weeks compared to placebo.
Figure 17 shows the percentage change in pH at 2 weeks, 6 weeks, 8 weeks and 12 weeks compared to placebo.
Figure 18 shows the percentage change in pH per dose for 2 weeks, 6 weeks, 8 weeks and 12 weeks compared with placebo.
Figure 19a shows a change in the visual evaluation of the quality color of 12 weeks from baseline in the modified intent to treat (MITT) population.
Figure 19b shows a change in visual assessment of vaginal epithelial integrity from baseline to 12 weeks in the modified treatment intention (MITT) population.
Figure 19c shows a change in visual assessment of vaginal epithelium thickness from baseline to 12 weeks in the modified treatment intention (MITT) population.
Figure 19d shows the change in visual assessment of vaginal secretion from baseline to 12 weeks in the modified Therapeutic Intent (MITT) population.
Figure 20a shows the correlation between the four visual assessments and the total sum of dyspareunia at 12 weeks in the Therapeutic Intent (ITT) group.
Figure 20b shows the correlation between the total of four visual assessments and vaginal dryness at 12 weeks in the Therapeutic Intent (ITT) population.
Figure 21 shows the adjusted estradiol serum concentration (pg / ml) estimated for 1 day (square) and 12 weeks (diamonds) for the four treatment arms.
Figure 22 shows the adjusted estradiol serum concentration (pg / ml) estimated at 14 days (square) and 12 weeks (diamonds) for the four treatment arms.
Figure 23 shows the estradiol plasma levels measured in subjects after the estrous cycle after the estradiol formulation compared to the plasma levels measured in the subjects after the estradiol formulation.
Figure 24 shows the mean change from baseline of the total FSFI score at 12 weeks.
Figure 25a shows the average change in individual FSFI lubrication scores from baseline to 12 weeks.
Figure 25b shows an average change in individual FSFI excitement scores from baseline to 12 weeks.
Figure 25c shows an average change in individual FSFI satisfaction scores from baseline to 12 weeks.
Figure 25d shows the mean change in individual FSFI libido scores from baseline to 12 weeks.
Figure 25E shows the mean change in scores of individual FSFI orgasms from baseline to 12 weeks.
26A shows an estradiol soft gel capsule having a wider end between fingers.
Figure 26 (b) shows the insertion of an oestradiol soft gel capsule at the osteointegrated position. The soft gel is inserted into the lower third of the vial by a smaller end-up.
Figure 26c shows the insertion of an estradiol soft gel capsule at a standing position. The soft gel is inserted into the lower third of the vial by a smaller end-up.

In the following detailed description of the embodiments of the present disclosure, reference is made to the accompanying drawings, in which like reference numerals indicate like elements, in which are shown by way of illustration specific embodiments in which the present disclosure may be practiced. It is to be understood that this embodiment is described in sufficient detail to enable those skilled in the art to practice the present disclosure and that other embodiments may be utilized and that other changes may be made without departing from the scope of the present disclosure . The following detailed description is, therefore, not to be taken in a limiting sense, and the scope of the present disclosure is defined only by the appended claims. The term " or " as used in this disclosure should be understood to be defined as a logical sum (i.e., and / or) and the term " It should not indicate an exclusive declaration of judgment unless it is otherwise clearly indicated, such as a word.

I. Definition

The term " active pharmaceutical ingredient " (" API ") as used herein refers to the active compound (s) used to formulate the drug product.

The term " co-administered " as used herein means that two or more drug products are administered simultaneously or sequentially on the same or different days.

The term " drug product " as used herein refers to at least one active pharmaceutical ingredient combined with at least one excipient and provided in a unit dosage form.

The term " sub-curve area "(" AUC ") refers to the amount of an active pharmaceutical ingredient by the change in blood concentration of the active pharmaceutical ingredient (e.g., estradiol or progesterone) Means the area under the defined curve. "AUC _0-∞ " is the area under the concentration-time curve extrapolated to infinity after dosing. "AUC _{0 -t} " is the area under the concentration-time curve from time 0 to time t after dose administration, where t is the last time by measurable concentration.

The term " _Cmax " refers to the maximum value of the blood concentration shown in the curve representing the change in the blood concentration of the active pharmaceutical ingredient (e.g., progesterone or estradiol) over time, or the metabolite of the active pharmaceutical ingredient.

The term " _Tmax " means the time it takes for the metabolite of an active pharmaceutical ingredient (e. G., Estradiol or progesterone) or active pharmaceutical ingredient to reach its maximum value.

The term "bioavailability" with the meaning as defined in 21 CFR § 320.1 (a) means the rate and extent to which the API or active ingredient or active moiety is absorbed from the drug product and made available at the site of action. For example, bioavailability can be measured as the amount of API in the blood (serum or plasma) as a function of time. Pharmacokinetic (PK) parameters such as AUC, _Cmax or _Tmax can be used to measure and evaluate bioavailability. For drug products not intended to be absorbed into the bloodstream, bioavailability may be assessed by measurements intended to reflect the rate and extent of API or active ingredient or active moiety becoming available at the site of action.

The term "bioequivalence", having the meaning as defined in 21 CFR § 320.1 (e), means that the API or active ingredient or active moiety in a pharmaceutical equivalent or pharmaceutical alternative is administered at the same molar volume under similar conditions in a suitably designed study ≪ / RTI > means that there is no significant difference in the rate and extent available at the site of drug action when administered. If there is an intentional difference in speed (e. G., In a given sustained release formulation), any pharmaceutical equivalent or substitute will be sufficient to ensure that the active ingredient or moiety from each product is available at the site of drug action If there is no significant difference, it can be considered to be bioequivalence. This applies only if the difference in rate at which the active ingredient or moiety becomes available at the site of drug action is intentionally reflected in the proposed labeling and is not essential for obtaining an effective body drug concentration in chronic use, Is considered to be meaningless. Indeed, the two products are considered to be bioequivalent when the AUC, C _max , or optionally the 90% confidence interval of T _max is within 80.00% to 125.00%.

The term " biological equivalence ", " body equivalence " or " natural ", as used herein with the hormones disclosed herein, refers to hormones that are consistent with the chemical structure and effects of those occurring naturally or endogenously in the human body. An exemplary natural estrogen is estradiol.

The term " biological equivalent hormone " or " body equivalent hormone " means an active pharmaceutical ingredient that is structurally identical to a hormone found naturally or endogenously in the human body (e.g., estradiol and progesterone).

The term "estradiol" refers to (17β) -estra-1,3,5 (10) -triene-3,17-diol. Estradiol is also called inter-compatible with 17? -Estradiol, estradiol or E2 and is endogenously found in the human body. As used herein, estradiol refers to the bioequivalence or bioequivalence form of estradiol found in the human body with the following structure: < RTI ID = 0.0 >

Estradiol is provided in anhydrous or semi-hydrate form. For purposes of this disclosure, anhydrous or semi-hydrate forms can be substituted for others by occupying the proportion of water according to well known and understood techniques or by lack of water.

The term " solubilized estradiol " means that the estradiol or a portion thereof is solubilized or dissolved in the solubilizing agent (s) or formulation described herein. Solubilized estradiol is about 80% solubilized, about 85% solubilized, about 90% solubilized, about 95% solubilized, about 96% solubilized, about 97% solubilized, about 98% solubilized, about 99% Solubilized, or about 100% solubilised estradiol. In some embodiments, estradiol is " fully solubilized " by all or substantially all of the estradiol that is solubilized or dissolved in the solubilizer. About 97% solubilized, about 98% solubilized, about 99% solubilized, or about 100% solubilized estradiol as a fully solubilized estradiol. Solubility can be expressed as mass fraction (w / w (%) also referred to as% by weight).

The term " progesterone " means preg-4-ene-3,20-dione. Progesterone is also called P4 and is inter-compatible and is found endogenous in the human body. As used herein, progesterone refers to the bioequivalence or bioequivalence forms of progesterone found in a human body having the following structure:

The term " solubilized progesterone " means that the progesterone, or a portion thereof, is solubilized or dissolved in the solubilizing agent (s) or formulation described herein. In some embodiments, progesterone is " partially solubilized " by a portion of the progesterone that is solubilized or dissolved in the solubilizer and a portion of the progesterone that is suspended in the solubilizer. The partially solubilized progesterone may be about 1% solubilized, about 5% solubilized, about 10% solubilized, about 15% solubilized, about 20% solubilized, about 30% solubilized, about 40% solubilized, % Solubilized, about 60% solubilized, about 70% solubilized, about 80% solubilized, about 85% solubilized, about 90% solubilized, or about 95% solubilized progesterone. In another embodiment, the progesterone is " fully solubilized " by all or substantially all progesterone solubilized or dissolved in the solubilizer. Completely solubilized progesterone may comprise about 97% solubilized, about 98% solubilized, about 99% solubilized, or about 100% solubilized progesterone. Solubility can be expressed as mass fraction (w / w (%) also referred to as% by weight).

The terms " undifferentiated progesterone " and " undifferentiated estradiol ", as used herein, include undifferentiated progesterone and undifferentiated estradiol having an X50 particle size value of less than about 15 microns or having an X90 particle size value of less than about 25 microns do. The term " X50 " means that half of the particles in the sample are smaller in diameter than the given number. For example, undifferentiated progesterone with an X50 of 5 microns means that half of the particles have a diameter of less than 5 microns for a given sample of undifferentiated progesterone. Similarly, the term " X90 " means that ninety percent (90%) of the particles in the sample are smaller in diameter than the given number.

The term " glyceride " is an ester of glycerol (1,2,3-propanetriol) having an acyl radical of a fatty acid, also known as acylglycerol. When only one position of the glycerol molecule is esterified by a fatty acid, a "monoglyceride" or "monoacylglycerol" is produced, and when two positions are esterified, the "diglyceride &Quot; or " diacylglycerol " is produced, and when all three positions of glycerol are esterified by fatty acids, "triglyceride" or "triacylglycerol" is produced. If all the esterified positions contain the same fatty acid, the glyceride is " simple " while the glyceride is " mixed " if the esterified position contains different fatty acids. The carbon of the glycerol backbone is referred to as sn-1, sn-2 and sn-3, sn-2 is the intermediate carbon, sn-1 and sn-3 are the end carbon of the glycerol backbone.

The term " solubilizer " means a substance or combination of substances that solubilize active pharmaceutical ingredients (e.g., estradiol or progesterone). For example, without limitation, suitable solubilizing agents include an intermediate chain oil, and other solvents and cosolvents that solubilize or dissolve the active pharmaceutical ingredient to a desirable extent. Solubilizing agents suitable for use in the formulations disclosed herein are pharmaceutical grade solubilizing agents (e. G., Pharmaceutical grades, medium chain oils). It will be understood by those skilled in the art that other excipients or ingredients may be added to or mixed with the solubilizing agent to enhance the solubility or the properties or performance of the resulting formulation. Examples of such excipients include, but are not limited to, surfactants, emulsifiers, thickeners, colorants, flavors, and the like. In some embodiments, the solubilizing agent is a medium chain oil, and in some other embodiments, the medium chain oil is combined with the co-solvent (s) or other excipient (s).

The term " intermediate chain " is used to describe the aliphatic chain length of a fatty acid-containing molecule. Refers to a fatty acid, fatty acid ester or fatty acid derivative containing a fatty acid aliphatic tail or a 6 (C6) to 14 (C14) carbon atom, a 8 (C8) to 12 (C12) ) To 10 (C10) carbon atoms.

The terms " intermediate chain fatty acid " and " intermediate chain fatty acid derivative " are used to describe a fatty acid or fatty acid derivative having an aliphatic tail (i.e., carbon chain) having 6 to 14 carbon atoms. Fatty acids consist of unbranched or branched aliphatic tails attached to carboxylic acid functional groups. Fatty acid derivatives include, for example, fatty acid esters and fatty acid containing molecules, including mono-, di-, and triglycerides, including, without limitation, components derived from fatty acids. Fatty acid derivatives also include fatty acid esters of ethylene or propylene glycol. The aliphatic tail can be saturated or unsaturated (i. E. Having one or more double bonds between carbon atoms). In some embodiments, the aliphatic tail is saturated (i.e., no double bonds between carbon atoms). The intermediate chain fatty acid or the intermediate chain fatty acid derivative is an aliphatic having 6 to 14 carbons including C6-C14, C6-C12, C8-C14, C8-C12, C6-C10, C8- And having a tail. Examples of the intermediate chain fatty acids include, without limitation, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, and derivatives thereof.

As used herein, the term " oil " is intended to include a starting material or a precursor thereof, including undifferentiated progesterone or undifferentiated estradiol as described herein, a biologically equivalent progesterone or estradiol, Means any pharmaceutically acceptable oil, especially a medium chain oil, specifically excluding peanut oil.

The term " intermediate chain oil " means an oil wherein the composition of the fatty acid fraction of the oil is substantially a middle chain (i.e. C6 to C14) fatty acid, i.e. the composition profile of the fatty acid in the oil is substantially intermediate. As used herein, " substantially " means that between 20% and 100% (including the upper and lower limits) of the fatty acid fraction of the oil is an intermediate chain fatty acid, i.e., an aliphatic tail having 6 to 14 carbons ) ≪ / RTI > In some embodiments, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65% About 75%, about 85%, about 90%, or about 95% are comprised of a medium chain fatty acid. As used herein, " predominantly " means that at least 50% of the fatty acid fraction of the oil is comprised of a fatty acid having an intermediate chain fatty acid, i.e., an aliphatic carbon chain having 6 to 14 carbon atoms. One of ordinary skill in the art will readily understand that the term "alkyl content" or "alkyl distribution" of the term oil may be used in place of the "fatty acid fraction" of the term oil in identifying any oil or solubilizing agent, Are used interchangeably herein. Thus, a medium chain oil suitable for use in the formulations disclosed herein is an intermediate chain oil in which the fatty acid fraction of the oil is substantially a medium chain fatty acid, or an alkyl chain having an alkyl distribution or alkyl distribution substantially in the middle chain alkyl (C6-C12 alkyl ). ≪ / RTI > It will be understood by those skilled in the art that the intermediate chain oil suitable for use in the formulations disclosed herein is a pharmaceutical grade (e.g., a pharmaceutical grade intermediate chain oil). Examples of intermediate chain oils are, for example and without limitation, medium chain fatty acids, medium chain fatty acid esters of glycerol (for example, mono-, di- and triglycerides), propylene glycol Medium chain fatty acid esters, medium chain fatty acid derivatives of polyethylene glycol, and combinations thereof.

The term " ECN " or " equivalent carbon number " means the sum of the number of carbon atoms in the fatty acid chain of an oil and may be used, for example, to identify oil as an intermediate chain oil or as a long chain oil. For example, trypalmitin (glycerol tripalmitate), which is a simple triglyceride containing three fatty acid chains of 16 carbon atoms, has an ECN of 3x16 = 48. Conversely, the triglycerides with ECN = 40 were 8, 16 and 16; 10, 14 and 16; 8, 14 and 18; Quot; mixed " fatty acid chain lengths, such as < / RTI > Natural oils are often " mixed " with respect to certain fatty acids, but tend not to contain both long chain and medium chain fatty acids in the same glycerol backbone. Thus, triglycerides, which are 21 to 42 ECNs, usually contain mainly medium chain fatty acids while triglycerides, which are ECN's of greater than 43, typically contain long chain fatty acids. For example, in USP, the ECN of corn oil triglycerides will be in the range of 51-54. Medium chain di-glycerides, which are 12-28 ECNs, usually contain mainly middle chain fat chains, while diglycerides that are 32 or more ECNs will typically contain mainly long-chain fatty acid tail. The monoglyceride will have an ECN that matches the chain length of the unique fatty acid chain. Thus, a monoglyceride, which is an ECN in the range of 6 to 14, will mainly contain a medium chain fatty acid, and a monoglyceride, which is an ECN of 16 or more, will mainly contain a long chain fatty acid.

The average ECN of the mid-chain triglyceride oil is typically 21 to 42. [ For example, as described in the US Pharmacopoeia (USP), the intermediate chain triglycerides have the following composition as the exemplary oil described in the table below: 3 * [(6 * 0.02) 0.25) + (12 * 0.02) + (14 * 0.01)] = 25.8.

The ECN of an exemplary intermediate chain triglyceride oil may also be expressed in the range of from 24.9 to 27.0 (depending on the range described in USP). For oils with mixed mono-, di-, and triglycerides, or mono- and di-fatty acid glycols, the ECN of the total oil is determined for each individual component (e.g., C8 monoglyceride, C8 diglyceride , C10 monoglyceride and C10 monoglyceride), and taking the sum of the relative percentages of the components multiplied by the ECN normalized with monoglyceride for each component. For example, the oils with C8 and C10 mono- and diglycerides listed in the table below have an ECN of 8.3 and are therefore a medium chain oil.

As indicated differently, the ECN can be calculated as the length of each chain in the composition multiplied by its relative percentage in oil: (8 * .85) + (10 * .15) = 8.3.

As used herein, the term " excipient " means a non-API component, such as a solubiliser, an antioxidant, an oil, a lubricant, and the like used to formulate a pharmaceutical product.

The term " patient " or " subject " means an individual to which a pharmaceutical composition is administered.

The term " pharmaceutical composition " means at least a pharmaceutical composition comprising a solubilizing agent and an estradiol. As used herein, a pharmaceutical composition is delivered, for example, through a suppository (i.e., a vaginal suppository) or is absorbed into vagina.

The term " progestin " means any natural or artificial substance having pharmacological properties similar to progesterone.

The terms " treat ", " treating ", and " treatment " refer to any indication of the success of treatment or alleviation of an injury, disease or condition, including any objective or subjective parameter, About; Reduction of symptoms, or impairment, disease or condition that makes the patient more tolerant; Slowing down the rate of degeneration or decay; Or improvement of a patient's physical or mental well-being. The treatment or alleviation of symptoms may be based on objective or subjective parameters, including the results of a physical examination, neuropsychological test or psychiatric evaluation.

The terms " atrophic vaginitis ", " vulva-vaginal atrophy ", " vaginal atrophy ", and " VVA " are used interchangeably herein. The molecular morphology of VVA is well known in the medical arts.

As used herein, " sexual dysfunction " means a condition that has one or more symptoms of difficulty during any one or more steps. Functional anomaly can prevent an individual from enjoying sexual activity. Non-limiting examples of sexual function symptoms include, but are not limited to, decreased sexual desire, decreased sexual pleasure, decreased sexual arousal and exaggeration, avoidance and repulsion of reproductive contact, inability to maintain or maintain excitement, and persistent or recurrent Delay, or absence thereof. Sexual function is lifelong (no effective behavior at all) or acquired (after a period of normal functioning); Limited to a general or predetermined situation or a given partner; And may be whole or part.

As used herein, " sex drive " means the frequency with which one desires to engage in sexual activity and / or the frequency involved in sexual activity, as recognized by the individual. Sexual arousal, as perceived by an individual, includes the frequency of sexual thoughts, the degree of joy in movies, books, music, etc., with a degree of joy or delight in thought and fantasy about sexual and / For example, one or more cognitive activities.

As used herein, " sexual arousal " means sexually excited frequency, how easily sexual arousal occurs and / or excitement is maintained, as perceived by an individual. Mentally, excitement can include factors such as desire for sexual activity and an increase in excretion associated with sexual activity. Physiologically, excitement may include increased blood flow to the reproductive organs leading to clitoral congestion and vaginal lubrication.

As used herein, " lubricating action " means wetting at or near the vagina before, during, or after sexual activity. Increased lubrication increases the frequency of lubrication; Reduction of difficulty in becoming lubrication; And / or reducing the difficulty of maintaining lubrication.

As used herein, "satisfaction" means one or more positive emotions (eg, fulfillment, achievement, bliss, etc.) associated with sexual activity or sexual intercourse. Satisfaction may include, for example, satisfaction with the occurrence of sexual arousal or orgasm, satisfaction with the amount of intimacy with the partner, and satisfaction with total sexual life.

As used herein, " orgasm " refers to the peak of sexual arousal characterized by a subjective experience of intense joy that usually manifests itself in vaginal contractions in women. The increase in orgasm may include an increase in the frequency, duration, and / or intensity of the orgasm in the subject. The increase in orgasm can also include a reduction in the difficulty of reaching the orgasm.

II. Introduction

A pharmaceutical composition comprising solubilized estradiol designed to be absorbed into the vagina is provided herein. The pharmaceutical compositions disclosed herein are designed to be absorbed locally and, for example, in vaginal or peripheral tissues and have a therapeutic effect thereon. Data demonstrating the efficacy of the disclosed pharmaceutical compositions, and methods relating to pharmaceutical compositions, are further disclosed herein. In general, the pharmaceutical compositions disclosed herein are useful in other indications caused by a decrease or lack of VVA, dyspareunia and estrogen.

Additional aspects and embodiments of the disclosure are directed to providing an increase in patient ease of use and minimizing the side effects of potentially inappropriate insertions, the incidence of vulvar-vaginal fungal infections due to use of estradiol products applied to other vaginals To minimize the incidence of vulvar-vaginal fungal infections; And an improved side effect profile (e.g., pruritus) compared to, for example, VAGIFEM (registered trademark) (estradiol vaginal tablet, Novo Nordisk, Princeton, NJ).

III. Pharmaceutical composition

function

According to an embodiment, the pharmaceutical compositions disclosed herein are non-alcoholic or substantially non-alcoholic. The provision of pharmaceutical compositions provides improved patient compliance due to improvements over previous offerings. According to an embodiment, the pharmaceutical compositions disclosed herein are encapsulated in a soft gelatin capsule, which improves comfort during use. According to an embodiment, the pharmaceutical composition is substantially liquid, which is more easily absorbed in vaginal tissue and is dispersed over a wider surface area of vaginal tissue.

Estradiol

According to an embodiment, the pharmaceutical compositions disclosed herein are for vaginal insertion of single or multiple unit dosage forms. According to an embodiment, the estradiol in the pharmaceutical composition is at least about 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84% 99%, 99%, or 100% solubilized in the presence of the solvent. According to an embodiment, when the estradiol is not 100% solubilized, the remaining estradiol is in its undifferentiated form or in another form in which the undifferentiated form is converted after administration, in a form which is capable of being absorbed by the body and having undifferentiated Crystalline) form.

According to an embodiment, all or part of the estradiol is solubilized in the solubilizer during the manufacturing process. In accordance with an embodiment, all or part of the estradiol is solubilized after administration (e.g., the undifferentiated portion where the estradiol is not 100% solubilized is solubilized in body fluids after administration). In accordance with an embodiment, the solubilizing agent taught herein with or without additional excipients other than solubilizers is liquid or semi-solid, since estradiol is solubilized. Estradiol is present at body temperature (average 37 ° C) and generally at the vaginal pH (ranging from 3.8 to 4.5 in healthy patients or 4.6 to 6.5 in VVA patients) to the extent that estradiol is not fully solubilized at the time of administration / Should be substantially solubilized.

According to an embodiment, estradiol may be added to the pharmaceutical compositions disclosed herein as estradiol, estradiol hemihydrate or other grade estradiol forms used in pharmaceutical compositions or formulations.

Depending on the embodiment, the estradiol dosage strength varies. The dosage strength of estradiol (or, for example, estradiol half-hydrate to the extent that the water content of the estradiol half-hydrate is occupied) is at least about 1 microgram (占 퐂 or 占 퐂) to at least about 50 占 퐂. Specific dosage forms include at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 23 μg, 24 μg, 25 μg, 26 μg, 27 μg, 28 μg, 29 μg, 30 μg, 31 μg, 16 μg, 17 μg, 18 μg, 19 μg, 20 μg, 21 μg, 43 μg, 44 μg, 45 μg, 46 μg, 47 μg, 48 μg, 33 μg, 34 μg, 35 μg, 36 μg, 37 μg, 38 μg, 39 μg, , 49 [mu] g or 50 [mu] g of estradiol. According to an embodiment, the pharmaceutical composition comprises at least about 2.5 [mu] g; 7.5 mu g, 12.5 mu g, 18.75 mu g of estradiol. According to an embodiment, the pharmaceutical composition may comprise from about 1 g to about 10 g, from 3 g to 7 g, from about 7.5 g to 12.5 g, from about 10 g to about 25 g, from about 1 g, from about 2.5 g, 27.5 占 퐂, about 7.5 占 퐂 to 22.5 占 퐂, 10 占 퐂 to 25 占 퐂 of estradiol. The lowest clinically effective dose of estradiol is used for the treatment of VVA and other indications described herein. In some embodiments, the dosage of estradiol is about 4 μg. In one embodiment, the dosage of estradiol is about 10 μg. In another embodiment, the dosage of estradiol is about 25 μg.

Solvent system

According to an embodiment, the solvent system for solubilizing estradiol is a medium chain fatty acid-based solvent together with other excipients. According to an embodiment, the solvent system comprises a non-toxic pharmaceutically acceptable solvent, co-solvent, surfactant, and other excipients suitable for vaginal delivery or absorption.

According to an embodiment, an oil having a medium chain fatty acid as a main component is used as a solubilizing agent for solubilizing estradiol. According to an embodiment, the solubilizing agent comprises a medium chain fatty acid ester (for example, an ester of glycerol, ethylene glycol or propylene glycol) or a mixture thereof. According to an embodiment, the middle chain fatty acid comprises a chain length of C6 to C14. According to an embodiment, the middle chain fatty acid comprises a chain length of C6 to C12. According to an embodiment, the middle chain fatty acid comprises a chain length of C8-C10. The ECN for the intermediate chain oil will range from 21 to 42 for triglycerides, 12 to 28 for diglycerides and 6 to 14 for monoglycerides.

According to an embodiment, the medium chain fatty acid is saturated. According to an embodiment, the intermediate chain fatty acids are predominantly saturated, i.e., greater than about 60% or greater than about 75%.

According to an embodiment, estradiol is soluble in the solubilizing agent at room temperature, but it may be desirable to warm the solubilizing agent during manufacture to improve viscosity. According to an embodiment, the solubilizer is a liquid between room temperature and about 50 占 폚, below 50 占 폚, below 40 占 폚, or below 30 占 폚.

According to an embodiment, the solubility of the estradiol in the medium chain oil, the mesophilic fatty acid, or the solubilizing agent (or oil / surfactant) is at least about 0.01 wt%, 0.02 wt%, 0.05 wt%, 0.06 wt%, 0.08 wt% , 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, 0.6 wt%, 0.7 wt%, 0.8 wt%, 0.9 wt%, 1.0 wt% or more.

According to an embodiment, the intermediate chain solubilizing agent includes, by way of example and without limitation, saturated, medium chain fatty acids: caproic acid (C6), enanthic acid (C7), caprylic acid (C8), pelargonic acid (C9) (C10), undecylic acid (C11), lauric acid (C12), tridecylic acid (C13) or myristic acid (C14). According to an embodiment, the solubilizing agent comprises an oil made from this free medium chain fatty acid, an oil of a medium chain fatty acid ester of glycerin, propylene glycol or ethylene glycol, or a combination thereof. This example mainly involves saturated mid-chain fatty acids (i.e., more than 50% of the fatty acids are medium chain saturated fatty acids). According to an embodiment, mainly C6 to C12 saturated fatty acids are contemplated. According to an embodiment, the solubilizing agent is selected from at least one of a solvent or a cosolvent.

According to an embodiment, the glycerin-based solubilizing agent includes mono-, di- or triglycerides, and combinations and derivatives thereof. Exemplary glycerin-based solubilizers include Miglyol TM, a caprylic / capry triglyceride (SASOL Germany GmbH, Hamburg, Germany). MIGLYOL is a mixture of Miglyol 810 (caprylic / caprylic triglyceride), Miglyol 812 (caprylic / capry triglyceride), Miglyol 816 (caprylic / capry triglyceride) and Miglyol 829 Acid / succinic acid triglyceride). For example, caproic acid / caprylic acid / caprylic acid / lauric acid triglyceride; Caprylic acid / caprylic acid / linoleic acid triglyceride; Other caprylic / capry triglyceride solubilizers including caprylic / caprylic / succinic acid triglycerides are likewise contemplated. According to an embodiment, CAPMUL MCM, medium chain mono- and di-glycerides are solubilizers. Other and triglycerides of fractionated vegetable fatty acids, and combinations or derivatives thereof, may be solubilizers according to embodiments. For example, the solubilizing agent may be saturated coconut oil and 1,2,3-propanetriol (glycerol, glycerin, glycerine) ester of palm kernel oil, and derivatives thereof.

Ethylene and propylene glycol (including polyethylene and polypropylene glycol) solubilizers include glyceryl mono- and di-caprylate; Propylene glycol monocaprylate (e.g., CAPMUL (R) PG-8 (CAPMUL brand is owned by ABITEC, Columbus, Ohio)); Propylene glycol monocaprate (e.g., CAPMUL PG-10); Propylene glycol mono- and dicaprylate; Propylene glycol mono- and dicaprate; Diethylene glycol monoester (e.g., TRANSCUTOL (R), 2- (2-ethoxyethoxy) ethanol, GATTEFOSSE SAS); And diethylene glycol monoethyl ether. Other combinations of mono- and di-esters of propylene glycol or ethylene glycol are clearly contemplated as solubilizing agents.

According to an embodiment, the solubilizing agent comprises a combination of mono- and di-propylene and ethylene glycol and a combination of mono-, di- and triglycerides. Depending on the embodiment, polyethylene glycol glyceride (Gelucire TM, GATTEFOSSE SAS (France Saint-Priest)) can be used herein as a solubilizer or as a surfactant. For example, GELUCIRE 44/14 (PEG-32 glyceryl laurate EP), a medium chain fatty acid ester of polyethylene glycol, is a mono-, di- and triglyceride and a mono- and diester of polyethylene glycol ≪ / RTI > polyethylene glycol glyceride.

According to embodiments, the commercially available fatty acid glycerol and glycol ester solubilizing agents may be comprised of ingredients other than fatty acid esters which are usually prepared from natural oils and thus primarily comprise or are characterized by solubilizing agents . These other ingredients include, for example, other fatty acid mono-, di-, and triglycerides; For example, fatty acid mono- and di-ester ethylene or propylene glycol, free glycerol or glycols, or free fatty acids. In some embodiments, when the oil / solubiliser is described herein as a saturated C ₈ fatty acid mono- or diastereomer of glycerol, the major components of the oil, i.e., greater than 50 wt% (e.g., greater than 75 wt% Greater than 85 wt%, or greater than 90 wt%) is caprylic monoglyceride and caprylic acid diglyceride. For example, the technical data sheet by ABITEC for CAPMUL MCM C8 describes CAPMUL MCM C8 composed of mono and diglycerides of a medium chain fatty acid (mainly caprylic acid) and has an alkyl content of at least 1% C6, 95 At least C8%, at least 5% C10 and at least 1.5% C12 and more.

For example, MIGLYOL 812 is a solubilizing agent generally described as C8-C10 triglyceride, as the fatty acid composition is at least about 80% triglyceride ester 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). The various MIGLYOL product information sheets illustrate various fatty acid components as follows:

According to an embodiment, the anionic or nonionic surfactant may be used in pharmaceutical compositions containing solubilized estradiol. The ratio of solubilizing agent (s) to surfactant (s) varies depending on the desired solubility of the respective solubilizing agent (s) and the respective surfactant (s) and the resulting pharmaceutical composition. For example, without limitation, CAPMUL MCM and nonionic surfactant may be used in proportions that include 65:35, 70:30, 75:25, 80:20, 85:15, and 90:10. Other non-limiting examples include, for example and without limitation, CAPMUL MCM and GELUCIRE 39/01 used in proportions comprising 6: 4, 7: 3, and 8: 2; For example, without limitation, CAPMUL MCM and GELUCIRE 43/01 used in proportions comprising 7: 3 and 8: 2; Examples include, without limitation, CAPMUL MCM and GELUCIRE 50/13 used in proportions that include 7: 3, 8: 2, and 9: 1.

Other excipients

According to an embodiment, the pharmaceutical composition further comprises a surfactant. The surfactant can be a nonionic surfactant, a cationic surfactant, an anionic surfactant, or a mixture thereof. Suitable surfactants include, for example, water-insoluble surfactants having a hydrophilic-lipophilic balance (HLB) value of less than 12 and water-soluble surfactants having an HLB value of greater than 12. Surfactants with high HLB and hydrophilicity help form oil-water droplets. Surfactants are both hydrophilic in nature and can dissolve or solubilize relatively large amounts of hydrophobic drug compounds.

Non-limiting examples include Tween, dimethylacetamide (DMA), dimethylsulfoxide (DMSO), ethanol, glycerin, N-methyl-2-pyrrolidone (NMP), PEG 300, PEG 400, Poloxamer 407 , Propylene glycol, phospholipids, hydrogenated soybean phosphatidylcholine (HSPC), diastereoyl phosphatidylglycerol (DSPG), L-α-dimyristoylphosphatidylcholine (DMPC), L-α-dimyristoylphosphatidyl (DMPG), polyoxyl 35 castor oil (CREMOPHOR EL, CREMOPHOR ELP), polyoxyl 40 hydrogenated castor oil (Cremophor RH 40), polyoxyl 60 hydrogenated castor oil (CREMOPHOR RH 60), polysorbate 20 (TWEEN 20), polysorbate 80 (TWEEN 80), d-? -Tocopheryl polyethylene glycol 1000 succinate (TPGS), Solutol HS-15, sorbitan monooleate (SPAN 20) / Capur glyceride (SOFTIGEN 767), PEG 400 capryl / kapur glyceride (LABRASOL), PEG 300 oleic acid glycine (LABRAFIL M-1944CS), polyoxyl 35 castor oil (ETOCAS 35), glyceryl caprylate (mono- and diglyceride) (IMWITOR), PEG 300 linoleic acid glyceride (LABRAFIL M- Polyoxyl 8 stearate (PEG 400 monostearate), polyoxyl 40 stearate (PEG 1750 monostearate), and combinations thereof. In addition, suitable surfactants include, for example, polyoxyethylene derivatives of sorbitan monolaurate, such as polysorbate, capryl caproylmorpholglyceride, polyglycolized glyceride, and the like.

According to an embodiment, the non-ionic surfactant may be at least one of glycerol and polyethylene glycol esters of long chain fatty acids, for example, lauroylmercrolol-32 glyceride or lauroylpolyoxyl-32 glyceride (Commercially available as GELUCIRE), e.g. GELUCIRE 39/01 (glycerol ester of saturated C12-C18 fatty acid), GELUCIRE 43/01 (ground NF / JPE) and GELUCIRE 50/13 -32 glyceride EP, stearoylpolyoxyl-32 glyceride NF, stearoylpolyoxyl glyceride (USA FDA IIG)). The surfactant may be used in various amounts, usually at a concentration of greater than about 0.01%, typically from about 0.01% to 10.0%, 10.1% to 20%, and 20.1% to 30%. In some embodiments, the surfactant can be used at a concentration of from about 1% to about 10% (e.g., from about 1% to about 5%, from about 2% to about 4%, from about 3% to about 8% .

According to embodiments, the nonionic surfactant includes, for example and without limitation, one or more of oleic acid, linoleic acid, palmitic acid and stearic acid. According to an embodiment, the nonionic surfactant comprises a polyethylene sorbitol ester comprising a commercially available polysorbate 80 under the trade name TWEEN TM 80 (Polysorbate 80) (Sigma Aldrich, St. Louis, Mo.) do. Polysorbate 80 contains approximately 60% to 70% oleic acid, the remainder mainly comprising linoleic acid, palmitic acid and stearic acid. Polysorbate 80 may be used in an amount of about 5 to 50% of the total mass of the pharmaceutical composition, depending on the embodiment, in an amount of about 30%.

According to an embodiment, the non-ionic surfactant comprises PEG-6 palmitostearate and ethylene glycol palmitostearate, which is commercially available as TEFOSE 63 (GATTEFOSSE SAS, France Saint-Priest) , Which may be used, for example, with a CAPMUL MCM having a ratio of MCM to TEFOSE 63 of 8: 2 or 9: 1, for example. According to an embodiment, the other solubilizer / nonionic surfactant combination includes, for example, Miglyol 812: GELUCIRE 50/13 or Miglyol 812: TEFOSE 63.

According to an embodiment, the surfactant can be an anionic surfactant, for example ammonium lauryl sulfate, dioctyl sodium sulfosuccinate, perfluoro-octanesulfonic acid, potassium lauryl sulfate or sodium stearate. Cationic surfactants are also contemplated.

According to embodiments, nonionic or anionic surfactants may be used alone with at least one solubilizing agent, or may be used in combination with other surfactants. Thus, such surfactants, or any other excipients as described herein, can be used to solubilize estradiol. The combination of solubilizer, surfactant and other excipients should be designed so that the estradiol is absorbed into the vaginal tissue. In accordance with an embodiment, the pharmaceutical composition will produce minimal vaginal discharge.

According to an embodiment, the pharmaceutical composition further comprises at least one thickener. In general, the thickening agent is added when the viscosity of the pharmaceutical composition causes a less desirable absorption. According to an embodiment, the surfactant (s) disclosed herein may also provide an increment of the pharmaceutical composition, which will help the estradiol to be absorbed by the vaginal mucosa upon release, while minimizing vaginal discharge. Examples of thickeners are: ground; Propylene glycol; A mixture of EP / NF / JPE, glyceryl lysine oleate, ethoxylated fatty alcohol (Ceteth-20, Steareth-20) EP / NF (OVUCIRE TM 3460 NF / JPE, glycerol monooleate (type 40) EP / NF / JPE, glycerol monooleate (type 40) EP / NF mixture (OVUCIRE WL 3264) A mixture of EP / NF (OVUCIRE WL 2944), a non-ionic surfactant comprising PEG-6 stearate, ethylene glycol palmitostearate and PEG-32 stearate, TEFOSE 63 or similar product, (WITEPSOL (R), Sasol Germany GmbH (Hamburg, Germany).) Other thickeners such as alginate, certain gums such as xanthan gum, agar-agar, iota carrageenan, kappa carrageenan and the like Some other compounds include thickeners such as gelatin, and HPMC, PVC and CMC. Depending on the embodiment, the viscosity of the pharmaceutical composition according to various embodiments may comprise from about 50 cps to about 1000 cps at 25 DEG C. Those skilled in the art will readily understand and choose suitable thickening agents .

According to an embodiment, the thickener is a nonionic surfactant. For example, polyethylene glycol saturated or unsaturated fatty acid esters or diesters are non-ionic surfactant thickeners. In an embodiment, the nonionic surfactant comprises a polyethylene glycol long chain (C16-C20) fatty acid ester and is selected from the group consisting of saturated or unsaturated C16-C18 fatty acids such as oleic acid, lauric acid, palmitic acid, Glycol long-chain fatty acid esters such as PEG-fatty acid esters or diesters. In an embodiment, the nonionic surfactant comprises a polyethylene glycol long chain saturated fatty acid ester and is selected from the group consisting of saturated fatty acid esters of saturated C16-C18 fatty acids, such as palmitic acid and stearic acid, Ethylene glycol-fatty acid ester. Such non-ionic surfactants may include PEG-6 stearate, ethylene glycol palmitostearate, and PEG-32 stearate such as, but not limited to, TEFOSE 63.

In accordance with an embodiment, TEFOSE 63 is used to provide additional viscosity and / or dispersibility in the vial to delay the flow of the composition from the vagina. Although the pharmaceutical composition is a liquid, the viscosity of such a pharmaceutical composition is such that the liquid remains in the API absorption area, so that the pharmaceutical composition is substantially absorbed by the tissue. Surprisingly, the addition of an excipient to increase the viscosity and / or dispersibility of the pharmaceutical composition herein permits the administration of a pharmaceutical composition that is liquid at body temperature, but is not excessively secreted from the vagina when the patient is standing, The patient is allowed to walk after administration of the composition.

According to the embodiment, the nonionic surfactant used as the thickener is not hydrophilic and has good emulsion properties. An illustrative example of such a surfactant is TEFOSE 63, which has a hydrophilic-lipophilic balance (HLB) value of about 9-10.

According to an embodiment, the pharmaceutical composition further comprises one or more mucoadhesive agents, for example, to increase the viscosity of the pharmaceutical composition to delay the flow discharge from the vagina, thereby improving the vaginal absorption of estradiol. According to another embodiment, alone or in addition to a change in viscosity, the mucoadhesive agent allows the pharmaceutical composition to chemically or mechanically adhere to the vaginal tissue. For example, the mucoadhesive agent may be present to assist the pharmaceutical composition by adhesion to the mucosa upon activation by water. According to an embodiment, the polycarbophil is a mucoadhesive. According to an embodiment, other mucoadhesive agents include, without limitation, poly (ethylene oxide) polymers having a molecular weight of from about 100,000 to about 900,000; Chitosan; A carbopol comprising a polymer of acrylic acid crosslinked with allyl cros or allylpentaerythritol; Polymers of C10-C30 alkyl acrylate and acrylic acid crosslinked by allylpentaerythritol; Copolymers containing carbomer homopolymers or block copolymers of polyethylene glycol and long chain alkyl acid esters; And the like. Depending on the embodiment, various hydrophilic polymers and hydrogels may be used as mucoadhesive. According to certain embodiments, the polymer or hydrogel may expand in response to contact with vaginal tissue or secretions, thereby enhancing the moisturizing and mucosal adhesive effects. The choice and amount of hydrophilic polymer may be based on the choice and amount of solubilizing agent. In some embodiments, the pharmaceutical composition comprises a hydrophilic polymer, but optionally excludes a gelling agent. In embodiments having a hydrogel, from about 5% to about 10% of the total mass may comprise a hydrophilic polymer. In a further embodiment, a hydrogel may be used. Hydrogels may include chitosan that expands in response to contact with water. In various embodiments, the cream pharmaceutical composition may comprise PEG-90M. In some embodiments, the mucoadhesive is present in a pharmaceutical formulation, a soft gel capsule, or both.

In accordance with an embodiment, the pharmaceutical composition may include, for example and without limitation, one or more thermoplastic (e.g., hydrophilic), hydrophilic monosaccharides containing hydrophilic sucrose and other saccharide-based ligands (US Pat. No. 6,018,033 thermoreversible gel.

According to an embodiment, the pharmaceutical composition further comprises a lubricant. In some embodiments, a lubricant may be present to assist in the manufacture of the formulation. For example, a lubricant may be added to ensure that the capsule or tablet does not adhere to each other during processing or storage. Any suitable lubricant may be used. For example, lecithin, a mixture of phospholipids, is a lubricant.

According to an embodiment, the pharmaceutical composition further comprises an antioxidant. Any suitable antioxidant may be used. For example, butylated hydroxytoluene, butylated hydroxy anisole and vitamin E TPGS.

According to an embodiment, the pharmaceutical composition comprises from about 20% to about 80% solubilizing agent, from about 0.1% to about 5% lubricant, and from about 0.01% to about 0.1% antioxidant.

The choice of excipient will depend on factors such as the effect of excipients on solubility and stability. Additional excipients used in various embodiments may include coloring agents and preservatives. Examples of colorants include FD & C colors (e.g., Blue No. 1 and Red No. 40), D & C colors (e.g., Yellow No. 10), and opacifiers (e.g., titanium dioxide). According to an embodiment, the colorant comprises from about 0.1% to about 2% by weight of the pharmaceutical composition. According to an embodiment, the preservative in the pharmaceutical composition comprises methyl and propyl paraben in a ratio of about 10: 1, in proportions of about 0.005% and 0.05% by weight.

In general, solubilizers, excipients, and other additives used in the pharmaceutical compositions described herein are non-toxic, pharmaceutically acceptable, compatible with each other, maintain stability of the various components and pharmaceutical compositions with respect to each other, . In addition, combinations of various components, including pharmaceutical compositions, will produce the desired therapeutic effect when administered to a subject.

Solubility of Estradiol

According to an embodiment, a mixture of intermediate chain fatty acid glycerides, for example C ₆ -C ₁₂ , C ₈ -C ₁₂ or C ₈ -C ₁₀ fatty acid mono- and diglycerides or mono-, di- and tri Solubilizers containing glycerides are dissolved in the estradiol. As illustrative examples, primarily C ₈ -C ₁₀ saturated fatty acid mono- obtained good results by the mixture of solubilizing agent, and the die-glycerides or medium-chain triglyceride (e.g., MIGLYOL 810 or 812). Longer chain glycerides do not appear to be suitable for dissolution of estradiol.

Solubilizers that include propylene glycol monocaprylate (e.g., capryol) and 2- (2-ethoxyethoxy) ethanol (e.g., TRANSCUTOL) also solubilize estradiol.

IV. Preparation of pharmaceutical compositions

According to an embodiment, the pharmaceutical composition comprises estradiol, for example, without limitation, at least one intermediate chain fatty acid, such as a medium chain fatty acid consisting of at least one mono-, di- or triglyceride, ≪ / RTI > or a pharmaceutically-acceptable solubilizing agent including any combination thereof. According to an embodiment, the pharmaceutical composition also comprises at least one glycule or a derivative thereof or a combination thereof or a combination of at least one glyceride and glycols. The glycol (s) may be used as a solubilizer or to adjust viscosity, and thus may be considered to be a thickener as further described herein. For example, without limitation, other excipients, including antioxidants, lubricants, and the like, are optionally added. According to an embodiment, the pharmaceutical composition comprises sufficient solubilizing agent to fully solubilize estradiol. However, it is clearly understood that other volumes of solubilizer may be used depending on the level of desired estradiol solubilization. One skilled in the art will know and understand how to determine the solubilizing agent and the capacity of other excipients according to the desired percentage of estradiol solubilized in the pharmaceutical composition.

In an exemplary embodiment, GELUCIRE 44/14 (Lauroyl Macrogol-32 Glyceride EP, Lauroyl Polyoxyl-32 Glyceride NF, Lauroyl Polyoxy Glyceride (USA FDA IIG)) CAPMUL MCM is heated to about 40 DEG C to facilitate mixing of oil and nonionic surfactant, but such heating is not necessary to dissolve estradiol.

The specific examples disclosed herein provide additional principles and embodiments illustrating the preparation of the pharmaceutical compositions disclosed herein.

V. Forwarding Vehicle

Generally, the pharmaceutical compositions described herein are delivered into the vagina within a delivery vehicle, such as a capsule. According to an embodiment, the capsule is a soft capsule of a material well known in the pharmaceutical art, for example, gelatin. However, according to an embodiment, the delivery vehicle is integrative with the pharmaceutical composition (i. E., The pharmaceutical composition is a delivery vehicle). In such embodiments, the pharmaceutical composition is a gel, cream, ointment, tablet, or other formulation that is directly applied or absorbed into the vagina.

According to an embodiment, the capsule does not contain at least one of a hydrophilic gel-forming bioadhesive, a lipophilic substance, a gelling agent for a lipophilic substance and / or a water-dispersible substance. According to an embodiment, the capsule does not contain a hydrophilic gel-forming bioadhesive selected from carboxyvinylic acid, hydroxypropylcellulose, carboxymethylcellulose, gelatin, xanthan gum, guar gum, aluminum silicate, and mixtures thereof. According to an embodiment, the capsules do not contain lipophilic substances selected from liquid triglycerides, solid triglycerides (with a melting point of about 35 占 폚), carnauba wax, cocoa butter, and mixtures thereof. According to an embodiment, the capsule does not contain a hydrophobic colloidal silica gelling agent. According to an embodiment, the capsule does not contain a water-dispersible material selected from polyoxyethylene glycol, polyoxyethylene glycol 7-glyceryl-cocoate, and mixtures thereof. In some embodiments, estradiol comprises a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; And a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate. In this embodiment, the hydrophilic gel-forming bioadhesive in the liquid composition. In some such embodiments, the liquid composition is contained by a gelatin capsule as described herein. In some of these embodiments, the capsules may contain one or more of gelatin and optionally gelatin, hydrolyzed gelatin, sorbitol-sorbitan solution, water, glycerin, titanium dioxide, FD & C Red No. 40, ethanol, ethyl acetate, propylene glycol, At least one additional component selected from the group consisting of acetate phthalate, isopropyl alcohol, polyethylene glycol, and ammonium hydroxide.

According to an embodiment, the delivery vehicle is designed for ease of insertion. According to an embodiment, the delivery vehicle is sized so that it can be inserted comfortably into the vagina. According to an embodiment, the delivery vehicle is manufactured in a variety of geometries. For example, the delivery vehicle is shaped into a teardrop, a cone with a frustum end, a cylinder with a larger " cap " portion, or other shape that fits the vagina and facilitates insertion into the vagina. According to an embodiment, a delivery vehicle is used in connection with an applicator. According to another embodiment, the delivery vehicle is inserted digitally.

According to an embodiment there is provided a method for the treatment of VVA comprising dyspepsia, vaginal dryness and estrogen deficiency urinary tract conditions (including urinary tract infections), wherein the composition for the treatment of VVA is administered at about 2 inches by vaginas, Inserted digitally at one-third of the query, generating at least one of the following: improved compliance compared to other products for the treatment of VVA; An improved user experience compared to other products for the treatment of VVA; And statistically significantly improved symptoms of VVA compared to placebo or baseline in one of the weeks 2, 4, 6, 8, 10 or 12 weeks or more after initiation of administration. According to an embodiment there is provided a method for the treatment of VVA, including dyspepsia, vaginal dryness and estrogen deficiency urinary tract conditions (including urinary tract infections), comprising administering a composition for the treatment of VVA and a delivery comprising teardrop shape The vehicle is inserted into the vagina about 2 inches or ⅓ of the vagina closest to the vaginal opening and produces at least one of the following: improved compliance compared to other products for the treatment of VVA; An improved user experience compared to other products for the treatment of VVA; And statistically significantly improved symptoms of VVA compared to placebo or baseline in one of the weeks 2, 4, 6, 8, 10 or 12 weeks or more after initiation of administration.

2, delivery vehicle 200 includes a pharmaceutical composition 202 and a capsule 204. In one embodiment, The width 208 represents the thickness of the capsule 204, e.g., about 0.108 inches. The distance from one end of the delivery vehicle 200 to another end is indicated as a distance 206, e.g., about 0.690 inches. The size of the delivery vehicle 200 may also be described by an arc past a radius of a given length. For example, the arc 210 defined by the outside of the gelatin 204 is an arc having a radius of about 0.189 inches. The arc 212 defined by the interior of the capsule 204 is an arc past a radius of about 0.0938 inches. The arc 214 defined by the outside of the gelatin 204, which is opposite to the arc 210, is an arc past a radius of about 0.108 inches. Suitable capsules of different dimensions may be provided. In accordance with an embodiment, the capsules 204 have dimensions that are the same or similar to those provided above for each other. In some embodiments, the gelatin capsules comprise at least one selected from the group consisting of hydrolysed gelatin, sorbitol-sorbitan solution, water, glycerin, titanium dioxide, FD & C Red No. 40, ethanol, ethyl acetate, propylene glycol, polyvinyl acetate phthalate, Alcohol, polyethylene glycol, and ammonium hydroxide.

According to an embodiment, the delivery vehicle is designed to remain in the vial until the pharmaceutical composition is released. According to an embodiment, the delivery vehicle dissolves in the vagina and is absorbed into the vaginal tissue by the pharmaceutical composition, which minimizes vaginal discharge. In this embodiment, the transfer mechanism is made from a non-toxic component, such as gelatin.

Design factors for vaginal insertion-type pharmaceutical compositions

In accordance with an embodiment, the pharmaceutical composition is designed to maximize good characteristics that result in patient compliance (patients who discontinue treatment prior to completion of a given treatment course) without sacrificing efficacy. Preferred features include, for example, a lack or reduction of stimulation compared to other hormone replacement patencies, a lack or elimination of vaginal discharge of pharmaceutical compositions and delivery vehicles in contrast to other hormone replacement patencies, Lack of delivery vehicle residues, ease of administration compared to other hormone replacement patencies, or improved efficacy of the drug product for other similar pharmaceutical compositions.

According to an embodiment, the pharmaceutical composition is non-irritating or minimizes irritation. Patient stimuli include pain, pruritus (itching), soreness, excessive secretion, swelling or other similar conditions. Patient stimulation produces poor compliance. Non-irritating or reduced irritant pharmaceutical compositions are measured against competing hormone paddies, including tablet, cream or other intranasal estrogen delivery forms.

According to an embodiment, the pharmaceutical composition does not cause systemic exposure (e. G., Blood circulation of estradiol), which improves safety. According to another embodiment, the pharmaceutical compositions disclosed herein significantly reduce systemic exposure (e. G., Blood circulation of estradiol) as compared to other vaginal dosing drugs marketed for the treatment of VVA.

In certain embodiments, the administration of the pharmaceutical composition comprises an average concentration (C _ave ) value of less than 20.6 pg / ml on one day of treatment, and / or a C _ave value of less than 19.4 pg / / RTI > and / or provides a C _ave value of less than 11.5 pg / ml on day 83 of treatment. In certain embodiments, the administration of the pharmaceutical composition comprises an average concentration (C _ave ) value of less than 10 pg / ml on one day of treatment, and / or a C _ave value of less than 7.3 pg / Or a C _ave value of less than 5.5 pg / ml on day 83 of treatment.

According to an embodiment, the pharmaceutical composition does not leave any residue inside the vagina. Rather, the pharmaceutical compositions and delivery vehicles are substantially absorbed or dispersed without causing unpleasant feel of unabsorbed residues or unabsorbed or undispersed drug products. Measurement of the absence of residues may be for other vaginal inserts or may be objectively measured by examination of vaginal tissue. For example, some other vaginally inserted products contain starch that can generate more secretions from the vagina after administration. In some embodiments, the pharmaceutical compositions provided herein provide a lesser amount, duration, or frequency of post-dose secretion compared to other vaginal inserts (e. G., Compressed tablets).

In accordance with an embodiment, the pharmaceutical composition improves vaginal discharge as compared to other paddle containing a hormone-carrying paddle. Ideally, vaginal discharge is eliminated or minimized or improved compared to the competition.

According to an embodiment, the pharmaceutical compositions disclosed herein are inserted digitally. According to an embodiment, the pharmaceutical composition is inserted digitally into the vagina approximately 2 inches without the need for an applicator. According to an embodiment, the pharmaceutical composition is designed to be inserted also by the applicator, if desired. According to some embodiments, because the site of VVA is in the proximal region of the vagina (toward the vaginal entrance), the pharmaceutical compositions disclosed herein are designed to be inserted at the proximal portion of the vagina.

Through extensive experimentation, the various intermediate chain fatty acid esters of glycerol and propylene glycol exhibit one or more favorable characteristics for developing as a human drug product. According to an embodiment, the solubilizing agent is selected from at least one of a solvent or a cosolvent. Suitable solvents and co-solvents include any mono-, di- or triglycerides and glycols, and combinations thereof.

According to an embodiment, the pharmaceutical composition is delivered through a gelatin capsule delivery vehicle. According to this embodiment, the pharmaceutical composition is a liquid pharmaceutical composition. According to an embodiment, the delivery vehicle is a soft capsule, for example a soft gelatin capsule. Thus, the pharmaceutical composition of this embodiment is encapsulated in a soft gelatin capsule or other soft capsule.

According to an embodiment, the pharmaceutical composition comprises a solubilizing agent comprising at least one C6 to C14 medium chain fatty acid mono-, di- or triglyceride and, optionally, at least about 80% solubilized estradiol in the thickener. According to an embodiment, the pharmaceutical composition comprises one or more C6 to C12 intermediate chain fatty acid mono-, di- or triglycerides, for example one or more C6 to C14 triglycerides, for example one or more C6 to C12 triglycerides , Such as at least about 80% solubilized estradiol in one or more C8-C10 triglycerides. This embodiment contemplates specifically estradiol that is at least 80% solubilized. This embodiment contemplates specifically estradiol that is at least 90% solubilized. This embodiment contemplates specifically estradiol that is at least 95% solubilized. This embodiment contemplates specifically fully resolved oestradiol.

As described above, the liquid pharmaceutical composition is a liquid at room temperature or at body temperature. For example, in some embodiments, the pharmaceutical composition provided herein is a liquid formulation contained within a soft gel capsule. Gels, other solid forms other than liquids at ambient or room temperature are less preferred for embodiments of pharmaceutical compositions that are liquid.

The thickener acts to increase the viscosity to, for example, less than about 10,000 cP (10,000 mPa.s), typically from about 5,000 cP or less, more typically about 50 to 1000 cP. In embodiments, nonionic surfactants, such as GELUCIRE or TEFOSE, may be solid at room temperature and require melting to be effectively mixed with the solubilizing agent. However, in this embodiment, the resulting pharmaceutical composition is in a liquid rather than a solid, even at higher viscosities.

According to an embodiment, the pharmaceutical composition comprises estradiol, a medium chain solubilizing agent and a thickening agent as ingredients delivered via a soft capsule delivery vehicle. Other ingredients such as coloring agents, antioxidants, preservatives or other ingredients may also be included. However, the addition of the other ingredients should be such that the solubility of the estradiol, the pharmacokinetics of the pharmaceutical composition or the efficacy of the pharmaceutical composition does not significantly change. Other factors that should be considered when adjusting the components of a pharmaceutical composition include stimulation, vaginal discharge, vaginal discharge and other related factors, such as reducing patient compliance. Other contemplated ingredients include oil or fatty acid esters, lecithin, mucoadhesives, gelling agents, dispersants, and the like.

VI. Way

According to an embodiment, the pharmaceutical compositions disclosed herein are useful for the treatment of at least one VVA condition, including vaginal dryness, vaginal or vulvar irritation or itching, dysuria, dyspareunia and vaginal bleeding associated with sexual activity, ≪ / RTI > According to an embodiment, the method of treatment is generally applicable to women.

According to embodiments, the pharmaceutical compositions disclosed herein may be used for the treatment of an estrogen-deficient urinary condition. According to an embodiment, the pharmaceutical compositions disclosed herein may be used for the treatment of vaginal bleeding associated with dyspareunia or sexual activity.

According to an embodiment, the treatment of vaginal bleeding associated with VVA, estrogen deficient urinary status, dyspareunia and sexual activity occurs by administering a pharmaceutical composition in vagina. According to an embodiment in which the delivery vehicle is capsule, the patient acquires the capsule, inserts the capsule into the vial where the capsule dissolves and the pharmaceutical composition is released into vagina where it is absorbed into the vaginal tissue. In some embodiments, the pharmaceutical composition is completely absorbed into vaginal tissue. In some embodiments, the pharmaceutical composition is substantially absorbed into vaginal tissue (e.g., at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97% , At least about 98 wt%, or at least about 99 wt% of the composition is absorbed). According to an embodiment, the capsule is inserted into the vagina at about 2 inches, but the depth of insertion is generally any depth that allows absorption of substantially all of the pharmaceutical composition. According to an embodiment, the capsule may also be applied by using an applicator that places the capsule at an appropriate vaginal depth as disclosed herein. According to an embodiment, the capsule is inserted into the lower third of the vial (i.e., one third closest to the vaginal opening). According to an embodiment, the soft gel capsule may be held by a larger end between the fingers as shown in Figure 26A.

The object will select the most comfortable position (e.g., a standing position as shown in Figure 26 (b) or a standing position as shown in Figure 26 (c)) and the subject will insert a soft gel something to do. Soft gel capsules will dissolve quickly. The soft gel can be inserted at any time of the day, and normal activity can be resumed immediately. According to an embodiment, the same time of day is used for all insertion of the soft gel.

In accordance with an embodiment wherein the pharmaceutical composition is a cream, gel, ointment or other similar formulation, the pharmaceutical composition is well known and applied digitally as understood in the art.

Upon release of the pharmaceutical composition in vagina, estradiol is absorbed locally. For example, a therapeutically effective concentration of estradiol is provided over a 24 hour period in the proximal region of the vagina after administration of the suppository to the proximal region of the patient's vagina.

Depending on the embodiment, the timing of administration of the pharmaceutical compositions of the present disclosure may be carried out by any safe means as prescribed by the practitioner. According to an embodiment, the patient will administer the pharmaceutical composition (e. G., Capsules) vaginally daily for 14 days and thereafter twice per week. In some such embodiments, the dose administered for two dosing periods per week is administered approximately three to four days apart. Typically, doses administered over two dosing periods per week do not exceed more than two doses per 7 day period.

According to an embodiment, the pharmaceutical composition may be an estrogen-based (or progestin- or progestin- and estrogen-based) pharmaceutical drug product orally administered, or a patch, cream, gel, spray, transdermal delivery system or other parenterally administered estrogen- Are administered to the vagina by simultaneous administration of the drug product, which may each comprise natural, biosimilar, or synthetic or other derived estrogens or progestins. In accordance with an embodiment, the adjustment of the circulating estrogen levels provided through administration of the pharmaceutical compositions disclosed herein is not intended to be additive to any concurrently administered estrogen product and its associated circulating blood levels. According to another embodiment, the co-administered estrogen product is intended to have a synergistic effect as determined by the patient's primary care physician.

According to an embodiment, a method of estrogenating vaginal tissue is provided. The method comprises administering (i. E. Suppository) or a dosage amount as described herein. Estrogenated vaginal tissue is typically present in the vaginal wall in the presence of clear secretions; Roughening and elasticity of the wall; Intact vaginal epithelium; And a characteristic of a pink tissue color. Conversely, de-estrogenated vaginal secretions are reduced or absent; Smooth tissue with little or no wrinkles; Bleeding of the vaginal surface; The occurrence of petechiae (i.e., the exact round points in the skin due to bleeding, which appear to be red, brown or purple); And pale or transparent tissue. Thus, estrogenizing vaginal tissue in accordance with the methods disclosed herein increases the level of vaginal secretion in a subject; Increasing the number of vaginal wrinkles in a subject; And / or reducing bleeding or petechiae in the subject. According to an embodiment, a method of estrogenating vaginal tissue is provided. The method comprises administering a suppository to provide an estradiol C _max or AUC as described herein. According to an embodiment, a method of estrogenating vaginal tissue is provided. The method comprises administering a suppository to provide an estron C _max or AUC as described herein.

According to an embodiment, methods for estrogenating the labia mina and the labia mina (collectively "labia") are provided as described herein. Generally, the pharmaceutical composition is inserted digitally at about 2 inches in the vial as shown in Figures 26a, 26b, and 26c, or at about 1/3 of the vial closest to the vaginal opening. Gelatin capsules containing pharmaceutical compositions dissolve, rupture or otherwise release the pharmaceutical composition into vials, so that the lower third of the labia and vagina are reestrogenated. According to some embodiments, the pharmaceutical composition is a liquid that flows partially in the labia and reestrogenates the labia directly.

According to an embodiment, a method of estrogenating the vulva is provided as described herein. Generally, the pharmaceutical composition is inserted digitally at about 2 inches in the vial as shown in Figures 26a, 26b, and 26c, or at about 1/3 of the vial closest to the vaginal opening. The gelatin capsules containing the pharmaceutical composition dissolve or rupture or otherwise release the pharmaceutical composition with vaginal fluid, so that both the vulva and the lower third of the vagina are reestrogenated. According to some embodiments, the pharmaceutical composition is a liquid that flows partially into the vulval tissue and reestrogenates the vulva directly.

According to an embodiment, a method of treating vaginal dryness is provided. The method includes administration of a soft gelatinous estradiol preparation (i. E. Suppository) or a dosage amount as described herein. The treatment of vaginal dryness according to the methods disclosed herein can be used to reduce the severity of vaginal dryness by 1%, 5%, 10%, 15%, 20%, 25%, 30%, 45%, 50%, 55% 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%. The reduction in severity may be achieved after 2 weeks of treatment, 6 weeks of treatment, 8 weeks of treatment, or 12 weeks of treatment. In some embodiments, the vaginal dryness is assessed using a severity scale ranging from 0 to 4 points, where 0 indicates no dryness, 1 indicates mild dryness, and 2 indicates normal dryness And 3 points indicate severe dryness.

In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 2 before treatment of the subject to 2 after 2 weeks treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness severity score from 2 before treatment of the subject to 1 after 2 weeks treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 0 before treatment of the subject to 0 after 2 weeks treatment of the subject.

In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 2 before treatment of the subject to 2 after 6 weeks treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness severity score from 2 before treatment of the subject to 1 after 6 weeks treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 0 before treatment of the subject to 0 after 6 weeks of treatment of the subject.

In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 2 before treatment of the subject to 2 after 8 weeks treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 2 prior to treatment of the subject to 1 after 8 weeks of treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 0 prior to treatment of the subject to 0 after 8 weeks of treatment of the subject.

In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 2 before treatment of the subject to 2 after 12 weeks of treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness severity score from 2 before treatment of the subject to 1 after 12 weeks treatment of the subject. In some embodiments, the method of treating vaginal dryness comprises reducing the dryness severity score from 0 before treatment of the subject to 0 after 12 weeks treatment of the subject.

In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness after two weeks of treatment, wherein the severity is assessed on a scale from 0 to 3, and the average decrease is from 0.5 to 1.25 . The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness after 6 weeks of treatment, wherein the severity is assessed on a scale from 0 to 3, and the average decrease is from 0.75 to 1.5 . The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness after 8 weeks of treatment, wherein the severity is assessed on a scale from 0 to 3, and the average decrease is from 0.9 to 1.5 . The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vaginal dryness comprises reducing the severity of dryness after 12 weeks of treatment, wherein the severity is assessed on a scale from 0 to 3, and the average decrease is from 0.9 to 1.5 . The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vaginal dryness comprises administering a suppository to provide an estradiol C _max or AUC as described herein. According to an embodiment, there is provided a method of treating vaginal dryness, the method comprising administering a suppository to provide an estron C _max or AUC as described herein.

According to an embodiment, a method of treating vulvar and / or vaginal itching or irritation is provided. The method includes administration of a soft gelatinous estradiol preparation (i. E. Suppository) or a dosage amount as described herein. Treatment of the vulvar and / or vaginal itching or irritation according to the methods disclosed herein may be used to reduce the severity of vulvar and / or vaginal itching or irritation by 1%, 5%, 10%, 15%, 20%, 25%, 30% , 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%. The reduction in severity may be achieved after 2 weeks of treatment, 6 weeks of treatment, 8 weeks of treatment, or 12 weeks of treatment. In some embodiments, vulvar and / or vagal itching or irritation is assessed using a severity scale ranging from 0 to 4, where 0 indicates no itching or irritation, and 1 indicates mild itching or irritation 2 indicates normal itching or irritation, and 3 indicates severe itching or irritation.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 2 before treatment of the subject to 2 after 2 weeks treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 2 before treatment of the subject to 1 after 2 weeks treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 0 before treatment of the subject to 0 after 2 weeks of treatment of the subject.

In some embodiments, a method of treating vulvar and / or vagal itching or irritation comprises reducing the itching / irritation severity score from 2 to 3 after 6 weeks of treatment of the subject prior to treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 1 before treatment of the subject to 1 after 6 weeks treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 0 prior to treatment of the subject to 0 after 6 weeks of treatment of the subject.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 2 prior to treatment of the subject to 2 after 8 weeks of treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 1 before treatment of the subject to 1 after 8 weeks treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 0 before treatment of the subject to 0 after 8 weeks of treatment of the subject.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 2 prior to treatment of the subject to 2 after 12 weeks of treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 1 prior to treatment of the subject to 1 after 12 weeks of treatment of the subject. In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the itching / irritation severity score from 0 before treatment of the subject to 0 after 12 weeks of treatment of the subject.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the severity of itching / irritation after 2 weeks of treatment, wherein the severity is assessed on a scale of 0 to 3 points, Is in the range of 0.3 point reduction to 0.6 point decrease. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the severity of itching / irritation 6 weeks after treatment, wherein the severity is assessed on a scale of 0 to 3 points, Is in the range of 0.5 point decrease to 0.7 point decrease. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the severity of itching / irritation 8 weeks after treatment, wherein the severity is assessed on a scale of 0 to 3 points, Is in the range of 0.5 point decrease to 0.8 point decrease. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises reducing the severity of itching / irritation after 12 weeks of treatment, wherein the severity is assessed on a scale of 0 to 3 points, Is in the range of 0.5 point decrease to 1.0 point decrease. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating vulvar and / or vaginal itching or irritation comprises administering a suppository to provide an estradiol C _max or AUC as described herein. According to an embodiment, there is provided a method of treating vulvar and / or vagus pruritus or irritation, the method comprising administering a suppository to provide an estron C _max or AUC as described herein.

According to an embodiment, a method of treating dyspareunia is provided. The method comprises administering or dosing the suppository as described herein. The treatment of dyspareunia according to the methods disclosed herein may be used to reduce the severity of dyspareunia by 1%, 5%, 10%, 15%, 20%, 25%, 30%, 45%, 50%, 55%, 60% , 70%, 75%, 80%, 85%, 90%, 95% or 99%. The reduction in severity may be achieved after 2 weeks of treatment, 6 weeks of treatment, 8 weeks of treatment, or 12 weeks of treatment. In some embodiments, dyspareunia is assessed using a severity scale ranging from 0 to 4, where 0 represents the absence of pain associated with sexual activity (by vaginal penetration) and 1 represents (by vaginal penetration) ) Indicates pain associated with mild sexual activity, 2 indicates pain associated with normal sexual activity (due to vaginal penetration), and 3 indicates pain associated with severe sexual activity (due to vaginal penetration).

In some embodiments, the method of treating dyspareunia includes reducing the dyspareunia severity score from 2 before treatment to 3 of the subject to 2 after 2 weeks treatment of the subject. In some embodiments, the method of treating dyspareunia includes reducing the dyspareunia severity score from 2 before treatment of the subject to 1 after 2 weeks treatment of the subject. In some embodiments, the method of treating dyspareunia comprises reducing the dyspareunia severity score from 0 before treatment of the subject to 0 after 2 weeks treatment of the subject.

In some embodiments, the method of treating dyspareunia comprises reducing the dyspareunia severity score from 2 prior to treatment of the subject to 2 after 6 weeks of treatment of the subject. In some embodiments, the method of treating dyspareunia comprises reducing the dyspareunia severity score from 1 before treatment of the subject to 1 after 6 weeks treatment of the subject. In some embodiments, the method of treating dyspareunia includes reducing the dyspareunia severity score from 0 before treatment of the subject to 0 after 6 weeks of treatment of the subject.

In some embodiments, the method of treating dyspareunia comprises reducing the dyspareunia severity score from 2 prior to treatment of the subject to 2 after 8 weeks of treatment of the subject. In some embodiments, the method of treating dyspareunia includes reducing the dyspareunia severity score from 1 before treatment of the subject to 1 after 8 weeks treatment of the subject. In some embodiments, the method of treating dyspareunia includes reducing the dyspareunia severity score from 0 before treatment of the subject to 0 after 8 weeks treatment of the subject.

In some embodiments, the method of treating dyspareunia comprises reducing the dyspareunia severity score from 2 before treatment of the subject to 2 after 12 weeks treatment of the subject. In some embodiments, the method of treating dyspareunia comprises reducing the dyspareunia severity score from 1 before treatment of the subject to 1 after 12 weeks treatment of the subject. In some embodiments, the method of treating dyspareunia includes reducing the dyspareunia severity score from 0 before treatment of the subject to 0 after 12 weeks treatment of the subject.

In some embodiments, the method of treating dyspareunia comprises reducing the severity of dyspareunia after 2 weeks of treatment, wherein the severity is assessed on a scale from 0 to 3, and the average reduction is from 0.9 to 1.1 Range. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating dyspareunia includes reducing the severity of dyspareunia 6 weeks after treatment, wherein the severity is assessed on a scale from 0 to 3, and the average reduction is from 1.3 to 1.5 Range. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating dyspareunia comprises reducing the severity of dyspareunia 8 weeks after treatment, wherein the severity is assessed on a scale from 0 to 3, and the average reduction is from 1.5 to 1.8 Range. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating dyspareunia comprises reducing the severity of dyspareunia 12 weeks after treatment, the severity is assessed on a scale of 0 to 3 points, and the average decrease is 1.5 to 1.8 Range. The average decrease may be determined by observing the number of any suitable object. In some embodiments, the number of subjects is at least 100. In some embodiments, the number of subjects is at least 500. In some embodiments, the number of subjects ranges from 700 to 800. In some embodiments, the number of subjects ranges from 740 to 750. In some embodiments, vaginal oestradiol preparation contains 4 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 10 g of estradiol. In some embodiments, the vaginal oestradiol preparation contains 25 g of estradiol.

In some embodiments, the method of treating dyspareunia comprises administering a suppository to provide an estradiol C _max or AUC as described herein. According to an embodiment, there is provided a method of treating dyspareunia, the method comprising administering a suppository to provide an estradiol C _max or AUC as described herein.

According to an embodiment, a method of treating a urinary tract infection is provided. The term "urinary tract infection" as used herein refers to a microorganism such as Escherichia coli , Staphylococcus saprophyticus , Klebsiella sp., Enterobacter sp. Refers to infection of the kidneys, ureters, bladder and urethra by Enterobacter sp. Or Proteus sp. Methods for treating urinary tract infections generally include administering a soft gelatinous estradiol formulation (i. E. Suppository) as described herein. According to certain embodiments, the method further comprises reducing urinary incontinence, urinary frequency or urination, hematuria, dysuria, and / or stress incontinence. According to certain embodiments, there is provided a method of treating a urinary tract infection comprising administering a suppository as described herein, and reducing the vaginal pH from greater than 4.5 to 3.5 to 4.5 (inclusive) do. The method may be particularly effective in treating urinary tract infections in elderly subjects (e. G., Over 65 years old, or over 75 years old, or over 85 years old). According to an embodiment, there is provided a method of treating a urinary tract infection, the method comprising administering a suppository to provide an estradiol C _max or AUC as described herein. According to an embodiment, there is provided a method of treating a urinary tract infection, the method comprising administering a suppository to provide an estron C _max or AUC as described herein. According to an embodiment, a method of treating sexual function is provided. The term " sexual dysfunction ", as used herein in connection with a female subject, generally refers to a pain or discomfort during sexual intercourse, a reduction in vaginal lubrication, a delay in vaginal congestion, an increase in time for excitement, , Decreased clitoral sensation, decreased sexual desire, and / or reduced excitability. According to an embodiment, there is provided a method of treating sexual function, said method comprising administering a suppository to provide an estradiol C _max or AUC as described herein. According to an embodiment, there is provided a method of treating sexual function, said method comprising administering a suppository to provide an estron C _max or AUC as described herein.

Sexual function and dysfunction may be assessed using the FSFI (Rosen R, Brown C, Heiman J, et al., "The Female Sexual Function Index (FSFI): A Multidimensional Self-Report Instrument for " Journal of Sex & Marital Therapy 2000. 26: p.191-208). FSFI is useful for assessing various domains of sexual functioning (eg, sexual desire, excitement, orgasm, satisfaction and pain). Thus, a method of treating sexual function as provided herein comprises the steps of administering a vaginal soft gel formulation to a subject and administering to the subject an effective amount of a compound selected from the group consisting of a full-scale FSFI score, an FSFI-libido score, an FSFI- Or increasing FSFI-orgasmic scores.

In some embodiments, the method of treating sexual function comprises administering estradiol to a subject and increasing the FSFI-libiter score by at least about 20%, or at least about 25%, or at least about 30%, compared to a baseline .

In some embodiments, the method of treating sexual function comprises administering estradiol to a subject and increasing the FSFI-excitatory score by at least about 30%, or at least about 40% or at least about 50%, compared to a baseline .

In some embodiments, the method of treating sexual function comprises administering estradiol to a subject and increasing FSFI-lubrication score by at least about 85% or at least about 95% or at least about 115% compared to a baseline do.

In some embodiments, the method of treating sexual function comprises administering estradiol to a subject and increasing the FSFI-orgasm score by at least about 40% or at least about 60% compared to a baseline.

In some embodiments, the method of treating sexual function comprises administering estradiol to a subject and increasing the overall FSFI score by at least about 50%, or at least about 55%, or at least about 70%, as compared to a baseline.

Examples of other measures for assessing sexual function include changes in sexual function ("CSFQ"; Clayton et al., Psychopharmacol Bull. 33 (4): 731-45 (1997) and Clayton et al., Psychopharmacol. 4): 747-53 (1997)); Interview with Derogatis on sexual function - self report ("DISF-SR"; Derogatis, J Sex Marital Ther . 23: 291-304 (1997)); Golombok-Rust List of Sexual Satisfaction ("GRISS"; Rust et al., Arch. Sex Behav . 15: 157-165 (1986)); Sexual Function Questionnaire ("SFQ"; Quirk et al., J Womens Health Gend Based Med . 11: 277-289 (2002)); And the Arizona Sex Experience Scale ("ASEX"; McGahuey et al., J Sex Marital Ther . 26: 25-40 (2000)), the entire disclosures of which are incorporated herein by reference. It does not. For assessments using questionnaires, measures of sexual function increase when the score in the appropriate domain, subscale, or subtest, as established for the questionnaire, indicates an abnormal sexual function. For example, a woman's sexual interest may be reduced when evaluated using the CSFQ when the subscale of the sexual interest score is less than 9. Conversely, sexual function is thought to be improved when the score in the appropriate domain, subscale, or subtest indicates higher (e. G., Normal or desired) sexual function. For clinical evaluation, sexual function may be assessed relative to a previous point in time for the patient for age, sex, sexual experience, and / or health and / or compared to the patient's peers, Lt; / RTI >

According to embodiments, the efficacy and safety of the pharmaceutical compositions described herein in the treatment of the symptoms of VVA can be determined. In accordance with an embodiment, the size, effect, cytology, histology, and variability of VVA may be used to determine the efficacy and safety of the pharmaceutical compositions described herein, or, as currently or further developed, as permitted in the art, Can be determined using various endpoints. One source of endpoints is the published guidelines of the US Food and Drug Administration (FDA) for the treatment of VVA by estradiol.

According to an embodiment, a gelatine capsule containing a pharmaceutical composition as disclosed herein may be administered in combination with a VVA (including a urinary tract infection) including dyspareunia, vaginal dryness and estrogen deficient urinary conditions (including urinary tract infections), which allows the subject to walk immediately or in a minute, Is provided. According to an embodiment, a gelatine capsule containing a pharmaceutical composition as disclosed herein may be formulated to contain a pharmaceutical composition at about 2 inches in vial as shown in Figures 26a, 26b and 26c, Lt; RTI ID = 0.0 > gelatin < / RTI > According to an embodiment, the gelatin capsules adhere to vaginal tissue shortly after being inserted into the vagina, dissolve, rupture or otherwise disintegrate, releasing the pharmaceutical composition. The pharmaceutical composition is dispersed in vaginal tissue and rapidly absorbed. According to an embodiment, the gelatin capsule is also completely absorbed by vaginal tissue. According to some embodiments, a viscosity enhancer, such as TEFOSE 63, provides an increased viscosity to allow the pharmaceutical composition to remain within the desired absorption site, thereby estrogenating the vagina, labia, and / or vulva. The combination of high viscosity, bioadhesion and rapid uptake prevents the need for the subject to be in the horn after administration so that the tissue absorbs estradiol, allowing the subject to walk immediately after or almost immediately after administration.

According to an embodiment, there is provided a method for treating VVA comprising dyspareunia, vaginal dryness and estrogen-deficient urinary status (including urinary tract infections) without causing non-naturally occurring secretions (e.g., secretion of the pharmaceutical composition or components thereof) / RTI > According to the above method, a soft gelatin capsule containing a liquid pharmaceutical composition that can be completely absorbed by vaginal tissue is administered. According to an embodiment, the pharmaceutical composition itself is completely absorbed by vaginal tissue. According to an embodiment, the pharmaceutical composition and the gelatin capsule are administered in a volume and size such that the vaginal tissue of the subject respectively completely absorbs the pharmaceutical composition. According to an embodiment, this absorption will occur simultaneously with the pedestrian's body. According to this method, the gelatin capsule and the liquid pharmaceutical composition are completely absorbed by the vaginal tissue, where only secretions that occur after estrogenation of the vagina are natural secretions that women experience before menopause. "Natural" vaginal discharge means a small amount of fluid that flows out of the vagina every day, leaving the old cells lined with vagina. Natural secretions are usually transparent or milky. Non-natural secretions may be larger than natural secretions, different from natural secretions or colors, or different secretions with different natural secretions and viscosities. Non-naturally occurring secretion may also mean the secretion (e.g., leakage) of the pharmaceutical composition from the vagina.

In accordance with an embodiment, a method is provided for treating VVA, including dyspareunia, vaginal dryness, and estrogen-deficient urinary conditions (including urinary tract infections), using a liquid pharmaceutical composition. According to this method, a soft gelatin capsule containing a liquid composition for treating VVA is provided to a subject. The subject inserts a soft gelatin capsule containing a liquid composition for treating VVA in vivo, either digitally or by means of an applicator, wherein the soft gelatin capsule dissolves, ruptures or degrades and the liquid composition is released into vaginal fluid. According to an embodiment, the liquid composition for treating VVA is the pharmaceutical composition disclosed herein. According to an embodiment, the subject inserts a gelatin capsule at about 2 inches in vagina or at about 1/3 of the vagina closest to the vaginal opening. According to an embodiment, the subject is immediately after or shortly after the administration.

According to an embodiment, there is provided herein a method for treating VVA, including dyspepsia, vaginal dryness and estrogen deficiency urinary tract conditions (including urinary tract infections), wherein the method comprises administering a composition for the treatment of VVA with vaginal 2 weeks Lt; RTI ID = 0.0 > VVA < / RTI > One of ordinary skill in the art will appreciate that improvements may be statistically evaluated as described herein, and that any improvement may be a statistically significant improvement. According to an embodiment, the composition for the treatment of VVA is a liquid pharmaceutical composition as disclosed herein. According to an embodiment, the composition for the treatment of VVA is a liquid containing 1 g to 25 g of estradiol. According to an embodiment, the method of administration is the method disclosed herein, including the insertion method shown in Figures 26A, 26B and 26C. According to embodiments, at 2 weeks of measurement, estradiol is not detected systemically using standard pharmaceutical pharmacokinetic parameters such as AUC and _Cmax .

According to an embodiment, there is provided herein a method for treating VVA, including dyspepsia, vaginal dryness and estrogen deficiency urinary tract conditions (including urinary tract infections), the method comprising administering a composition for the treatment of VVA with vaginal 4 weeks Lt; RTI ID = 0.0 > VVA < / RTI > One of ordinary skill in the art will appreciate that improvements may be statistically evaluated as described herein, and that any improvement may be a statistically significant improvement. According to an embodiment, the composition for the treatment of VVA is a liquid pharmaceutical composition as disclosed herein. According to an embodiment, the composition for the treatment of VVA is a liquid containing 1 g to 25 g of estradiol. According to an embodiment, the method of administration is the method disclosed herein, including the insertion method shown in Figures 26A, 26B and 26C. According to embodiments, estradiol is not systemically detected using standard pharmaceutical pharmacokinetic parameters, such as AUC and _Cmax , at 2 weeks and / or 4 weeks of measurement.

According to an embodiment, there is provided herein a method for treating VVA, including dyspepsia, vaginal dryness and estrogen deficiency urinary tract conditions (including urinary tract infections), the method comprising administering a composition for the treatment of VVA with vix Lt; RTI ID = 0.0 > VVA < / RTI > One of ordinary skill in the art will appreciate that improvements may be statistically evaluated as described herein, and that any improvement may be a statistically significant improvement. According to an embodiment, the composition for the treatment of VVA is a liquid pharmaceutical composition as disclosed herein. According to an embodiment, the composition for the treatment of VVA is a liquid containing 1 g to 25 g of estradiol. According to an embodiment, the method of administration is the method disclosed herein, including the insertion method shown in Figures 26A, 26B and 26C. According to embodiments, estradiol is not detected systemically using standard pharmaceutical pharmacokinetic parameters, such as AUC and _Cmax , at 2 weeks and / or 8 weeks of measurement.

According to an embodiment, there is provided herein a method for treating VVA, including dyspepsia, vaginal dryness and estrogen deficiency urinary tract conditions (including urinary tract infections), which method comprises administering a composition for the treatment of VVA with vinegar for 10 weeks Lt; RTI ID = 0.0 > VVA < / RTI > One of ordinary skill in the art will appreciate that improvements may be statistically evaluated as described herein, and that any improvement may be a statistically significant improvement. According to an embodiment, the composition for the treatment of VVA is a liquid pharmaceutical composition as disclosed herein. According to an embodiment, the composition for the treatment of VVA is a liquid containing 1 g to 25 g of estradiol. According to an embodiment, the method of administration is the method disclosed herein, including the insertion method shown in Figures 26A, 26B and 26C. According to embodiments, estradiol is not detected systemically using standard pharmaceutical pharmacokinetic parameters such as AUC and _Cmax at 2 weeks and / or 10 weeks of measurement.

According to an embodiment, there is provided a method for treating VVA comprising dyspepsia, vaginal dryness and an estrogen-deficient urinary state (including urinary tract infections), said method comprising administering a therapeutically effective amount of VVA at any of two, four, six, At least one of the symptoms of VVA is compared to a baseline or placebo, wherein estradiol is not detected systemically using standard pharmaceutical pharmacokinetic parameters such as AUC and _Cmax . One of ordinary skill in the art will appreciate that improvements may be statistically evaluated as described herein, and that any improvement may be a statistically significant improvement. According to an embodiment, the composition comprising estradiol is a liquid composition as disclosed herein. According to an embodiment, the composition contains 1 g to 25 g of estradiol.

According to an embodiment, there is provided a method of reestrogenating a vagina, labia or vulva, the method comprising administering a composition comprising estradiol for the treatment of VVA, said composition comprising estradiol or synthetic estrogen And the liquid is dispersed over the surface area of the vagina, the labia or the vulva, which is wider than the area covered by the solid composition. For example, the liquid may have a viscosity of from about 50 cm2 to about 120 cm2 (e.g., from about 50 cm2 to about 60 cm2; or from about 60 cm2 to about 70 cm2; or from about 70 cm2 to about 80 cm2; Or from about 90 cm 2 to about 100 cm 2; or from about 100 cm 2 to about 110 cm 2; or from about 110 cm 2 to about 120 cm 2; or from about 65 cm 2 to about 110 cm 2). According to an embodiment, the subject inserts the liquid composition into the vagina with a capsule, for example a hard or soft gelatin capsule, which then dissolves in the vagina, ruptures or disintegrates or otherwise releases liquid. According to an embodiment, the liquid may contain at least one of a bioadhesive or a viscosity enhancing agent to prevent the liquid from being released from the vagina before the estradiol or synthetic estrogen can be absorbed into the vaginal tissue at a sufficient capacity to effect vaginal re- estrogenation . Depending on the embodiment, the quality will be reestrogenated statistically significantly within two weeks of administration compared to baseline or placebo levels. According to the embodiment, the quality will be reestrogenated statistically significantly within 4 weeks of administration compared to baseline or placebo levels.

According to the embodiment, the quality will be reestrogenated statistically significantly within 6 weeks of administration compared to baseline or placebo levels. According to the embodiment, the quality will be reestrogenated statistically significantly within 8 weeks of administration compared to baseline or placebo levels. According to the embodiment, the quality will be reestrogenated statistically significantly within 10 weeks of administration compared to baseline or placebo levels. According to an embodiment, the quality will be reestrogenated statistically significantly within 12 weeks of administration compared to baseline or placebo levels.

VII. Measurement of efficacy

According to an embodiment, administration of the pharmaceutical compositions described herein results in the treatment of VVA, and improvement of one or more of the associated symptoms. Patients with VVA experience shrinkage of the vasculature in both length and diameter, and the vasculature has fewer glycogen-rich vaginal cells to maintain moisture and suppleness. In addition, the vaginal wall may be thin, thin, dry, or sometimes inflamed (atrophic vaginitis). This change may be manifested by a variety of symptoms, collectively referred to as VVA. Such symptoms include, without limitation, increased vaginal pH; Reduced vaginal epithelium integrity, vaginal secretion, or epithelial surface thickness; Pruritus; Vaginal dryness; Dyspareunia (pain or bleeding during sexual intercourse); Urinary tract infections; Or a change in quality color. According to an embodiment, the efficacy is again measured as a reduction in vulvar and vaginal atrophy in the patient to a premenopausal state. According to an embodiment, the change is measured as a reduction in the severity of one or more atrophic effects compared to measurements taken at baseline (screening, 1 day) and taken 15 days (end of treatment). The severity of the atrophic effect can be measured using a scale of 0 to 3, where for example no = 0, mild = 1, moderate = 2 or severe = 3. This scoring is above all a suitable course of treatment regimen, Pre-treatment status and post-treatment outcome of the patient to determine dosage, frequency and duration of administration.

One of the symptoms of VVA is an increase in vaginal pH. In a further aspect of the disclosure, treatment with the pharmaceutical compositions described herein reduces vaginal pH. The decrease in vaginal pH is measured as the decrease from the vaginal pH at the reference value (screening) to the vaginal pH at 15 days. According to an embodiment. In some embodiments, a pH of greater than or equal to 5 may be associated with VVA. In some embodiments, the pH is measured using a pH indicator strip disposed against the vaginal wall. In some embodiments, the change in vaginal pH is a change in the vaginal pH of the patient to a pH of less than about pH 5.0. In some embodiments, the vaginal pH of the subject is less than about pH 4.9, pH 4.8, pH 4.7, pH 4.6, pH 4.5, pH 4.4, pH 4.3, pH 4.2, pH 4.1, pH 4.0, pH 3.9, , pH 3.6, or pH 3.5.

According to embodiments, treatment with the pharmaceutical compositions described herein improves the quality maturity index. Maturation index is measured as a change in cell composition. As with VVA, depending on the embodiment, the change in cell composition is dependent on a change in the composition or percentage of the amount of vaginal vaginal cells, mesenchymal cells and epidermal vaginal cells, such as changes in epidermal vaginal cells, Or by changes in the composition or amount of vaginal cells. Subjects with VVA symptoms usually have an increased number of ventricular cells and a decrease in the number of epidermal cells (for example, less than about 5%) compared to women who have not undergone VVA. Conversely, a subject with reduced VVA symptoms or otherwise responding to treatment may exhibit an improvement in maturation index, specifically a decrease in the amount of vesicular cells compared to a baseline (screening) or an increase in the amount of epidermal cells. In an embodiment, the decrease in basophil cells is measured as a decrease in the percentage of basal cells, and the percent reduction is at least about 85%, 80%, 75%, 70%, 65%, 60% , 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15% or 10%. In an embodiment, the percent reduction may be at least about a 54% reduction in the number of basal cells. In an embodiment, the increase in epidermal cells is measured as an increase in the percentage of epidermal cells, and the percentage increase in epidermal cells is at least about 5%, 10%, 15%, 20%, 25% , 30%, 35%, 40%, 45%, or 50% increase. In a further embodiment, the percent increase may be at least about 35% increase in the number of epidermal cells.

In some embodiments, the improvement in maturity score is assessed as a change over time. For example, as a change in cell composition measured at baseline (screening) on day 1 compared to the cell composition measured at day 15. Changes in cell composition can also be assessed as a change in the amount of proliferating cells over time, in addition to measuring changes in basal and epidermal cells as described above, optionally. These cells can be obtained from the vaginal mucosal epithelium through routine non-reproductive scientific testing and can be examined by vaginal smear.

In various further aspects of the disclosure, treatment with the pharmaceutical compositions described herein increases the number of epidermal cells; Reduction of ventricular basal cells; And an increase in intermediate cells.

In a further aspect of the disclosure, the sample may be collected to determine hormone levels, particularly estradiol levels. In some embodiments, a blood sample can be taken from the values of the subject and the measured estradiol (pg / ml). In some embodiments, estradiol levels can be measured at 0 hour (e.g., at the time of the first treatment), 1 hour (e.g., after the first treatment), 3 hours, and 6 hours. In some embodiments, the sample can be taken 8 days (e.g., after the first treatment) and 15 days (e.g., 1 day after the last treatment on the 14th day). In some embodiments, the descriptive statistics and observed C _max and T _max values of plasma estradiol concentrations at each sampling time are measured and the AUC can be calculated.

In some embodiments, the suppository may comprise about 25 μg of estradiol. In this case, the administration of the suppository to the patient can be effected in a plasma sample from the patient by: 1) administering a dose of about 19 pg * hr / ml to about 29 pg * hr / ml (e.g., about 19.55 pg * hr / * hr / ml) corrected geometric mean peak plasma concentration (C _max ) of estradiol; Or 2) a corrected curved geometric mean area (AUC) of _0-24 of estradiol of about 75 pg * hr / ml to about 112 pg * hr / ml (e.g., about 75.82 pg * hr / And may provide parameters including one or more selected parameters. In some embodiments, the administration of a suppository to a patient comprises: 1) administering a therapeutically effective amount of a compound of the invention in a plasma sample from a patient comprising 1) about 9 pg * hr / ml to about 14 pg * hr / ml (e.g., about 9.17 pg * hr / corrected peak-to-peak plasma concentration ( _Cmax ) of the estrone of pg * hr / ml; And 2) a corrected curve geometric mean area (AUC) of the estrone of from about 43 pg * hr / ml to about 65 pg * hr / ml (e.g., about 43.56 pg * hr / ml to about 64.06 pg * hr / _0.0 > _0-24 . &_{Lt; /} RTI > In some embodiments, administration of a suppository to a patient comprises: 1) administering a plasma sample from a patient comprising: 1) about 416 pg * hr / ml to about 613 pg * hr / ml (e.g., about 416.53 pg * hr / pg * hr / ml) corrected peak-to-peak plasma concentration (C _max ) of estrone sulfate; And 2) a corrected curve geometric mean area (AUC) of the estrone sulfate of from about 3598 pg * hr / ml to about 5291 pg * hr / ml (e.g., about 3598.04 pg * hr / ml to about 5291.24 pg * hr / ) _0-24 . &_{Lt; /} RTI >

In some embodiments, the suppository comprises about 25 μg of estradiol. In some such embodiments, the administration of the suppository to the patient comprises the following steps: 1) in a plasma sample from the patient: 1) about 20.9 pg / ml to about 32.8 pg / ml (e.g., about 20.96 pg / ml to about 32.75 pg / ) Corrected geometric mean peak plasma concentration (C _max ) of estradiol; 2) a corrected curve geometric mean of the estradiol ranging from about 104.3 pg * hr / mL to about 163.1 pg * hr / mL (e.g., about 104.32 pg * hr / mL to about 163.0 pg * hr / Area (AUC) _0-24 ; And 3) an average concentration of estradiol (C _avg ) in the range of about 4.3 pg / ml to about 6.8 pg / ml (e.g., about 4.32 pg / ml to about 6.75 pg / ml) Lt; RTI ID = 0.0 > a < / RTI >

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) a corrected geometric mean peak plasma concentration of estradiol of about 26.2 pg / ml (C _max ); 2) a corrected curve geometric mean area (AUC) of about 130 pg * hr / ml of estradiol _0-24 ; And 3) an average concentration of estradiol (C _avg ) of about 5.4 pg / ml when assessed on day 1.

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) about 9.5 pg / ml to about 15.1 pg / ml (eg, about 9.60 pg ( _Cmax ) of estradiol in the range of from about 0.001 mg / hr / ml to about 15.00 pg / ml); 2) a corrected curve geometric mean of the estradiol ranging from about 67.6 pg * hr / ml to about 105.8 pg * hr / ml (e.g., about 67.68 pg * hr / ml to about 105.75 pg * hr / Area (AUC) _0-24 ; And 3) an average concentration of estradiol in the range of about 2.7 pg / ml to about 4.4 pg / ml (e.g., about 2.80 pg / ml to about 4.38 pg / ml) of estradiol C _avg ). &_Lt; / RTI >

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) a corrected geometric mean peak plasma concentration of estradiol of about 12.0 pg / ml (C _max ); 2) a corrected curve geometric mean area (AUC) of about 84.6 pg * hr / ml of estradiol _0-24 ; And 3) an average concentration of estradiol (C _avg ) of about 3.5 pg / ml when evaluated at 14 days.

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) about 158.8 pg / ml to about 248.3 pg / ml (eg, about 158.88 hrs / Ml to about 248.25 pg * hr / ml); a corrected geometric mean peak plasma concentration (C _max ) of the estron conjugate; And 2) an estrone in the range of about 1963.1 pg * hr / ml to about 3067.6 pg * hr / ml (e.g., about 1963.20 pg * hr / ml to about 3067.50 pg * hr / Provides one or more parameters selected from the geometric mean area under the corrected curve (AUC) _0-24 of the conjugate.

In some embodiments, administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) a corrected geometric mean peak plasma concentration of the estron conjugate of about 198.6 pg / ml (C _max ); And 2) a corrected curve geometric mean area (AUC) _0-24 of the estrone conjugate of about 2454 pg * hr / ml when evaluated on day 1.

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) a plasma sample from the patient having 1) about 173.5 pg * hr / ml to about 271.3 pg * hr / ㎖ (e. g., about 173.60pg * hr / ㎖ to about 271.25pg * hr / ㎖; or about 217pg * hr / ㎖) unregulated curve ₀ arithmetic average area (AUC) in the range of estradiol of the diol _-24 ; 2) Estradiol in the range of about 7.2 pg / ml to about 11.4 pg / ml (e.g., about 7.25 pg / ml to about 11.33 pg / ml; or about 9.06 pg / ml) Corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) from about 137.5 pg * hr / ml to about 215.1 pg * hr / ml (e.g., from about 137.60 pg * hr / ml to about 215.00 pg * hr / ml, or about 172 pg * hr 0.0 > (AUC) < / RTI > of the estradiol in the range of _0-24 ; And 4) an amount of estradiol in the range of about 5.7 pg / ml to about 9.0 pg / ml (e.g., about 5.72 pg / ml to about 8.94 pg / ml; or about 7.15 pg / (C _{avg [0-24]} ). &_Lt; / RTI >

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in 1) at least about 1 mg / hr / ㎖ (e. g., about 335.20pg * hr / ㎖ to about 523.75pg * hr / ㎖; or about 419pg * hr / ㎖) the range of estrone ratio adjustment of the curve _O-arithmetic mean area (AUC) ₂₄ ; 2) Calibration of estrone in the range of about 13.9 pg / ml to about 21.9 pg / ml (e.g., about 14.00 pg / ml to about 21.88 pg / ml; or about 17.5 pg / ml) The arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) about 343.1 pg * hr / ml to about 536.2 pg * hr / ml (e.g., about 343.20 pg * hr / ml to about 536.25 pg * hr / ml, or about 429 pg * hr / 0.0 > (AUC) < / RTI >_0-24; And 4) a concentration of estrone in the range of about 14.3 pg / ml to about 22.4 pg / ml (e.g., about 14.32 pg / ml to about 22.38 pg / ml; or about 17.9 pg / And one or more parameters selected from a calibrated arithmetic mean peak plasma concentration (C _{avg [0-24]} ).

In some embodiments, the administration of a suppository comprising about 25 μg of estradiol to the patient results in a plasma sample from the patient having: 1) about 7,300.7 pg * hr / ml to about 11,407.6 pg * (AUC) of the estrone conjugate in the range of about 1 mg / hr / ml (e.g., about 7,300.80 pg * hr / ml to about 11,407.50 pg * hr / ml; or about 9,126 pg * hr / _0-24 ; 2) Calibration of the estron conjugate in the range of about 303.9 pg / ml to about 475.1 pg / ml (e.g., about 304.00 pg / ml to about 475.00 pg / ml; or about 380 pg / ml) The arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) about 7,943.9 pg * hr / ml to about 12,412.6 pg * hr / ml (e.g., about 7,944.00 pg * hr / ml to about 12,412.50 pg * hr / hr / ml) of the estron conjugate in the range of _0-24 ; And 4) an estrogen conjugate in the range of about 331.1 pg / ml to about 517.4 pg / ml (e.g., about 331.20 pg / ml to about 517.50 pg / ml; or about 414 pg / ml) And one or more parameters selected from a calibrated arithmetic mean peak plasma concentration (C _{avg [0-24]} ).

In some embodiments, the suppository may comprise about 10 μg of estradiol. In such a case, the administration of the suppository to the patient can be effected in a plasma sample from the patient by: 1) administering a dose of about 12 pg * hr / ml to about 18 pg * hr / ml (e.g., about 12.22 pg * hr / ml to about 17.98 pg * hr / ml) corrected geometric mean peak plasma concentration (C _max ) of estradiol; 2) a corrected curve geometric mean area (AUC) of estradiol of about 42 pg * hr / ml to about 63 pg * hr / ml (e.g., about 42.18 pg * hr / ml to about 62.02 pg * hr / _0-24 ; And 3) a corrected geometric mean time to peak plasma concentration of estradiol (T _max ) of from about 1 hour to about 3 hours (e.g., from about 1.49 hours to about 2.19 hours) . In some embodiments, the administration of the suppository to the patient comprises: 1) administering the plasma sample from the patient in an amount of about 4 pg * hr / ml to about 7 pg * hr / ml (e.g., about 4.38 pg * hr / corrected peak-to-peak plasma concentration ( _Cmax ) of the estrone of pg * hr / ml; 2) from about 20pg * hr / ㎖ to about 31pg * hr / ㎖ (e. G., About 20.60pg * hr / ㎖ to about 30.30pg * hr / ㎖) the correction curve and the geometric mean area (AUC) of estrone _{0 -24} ; And 3) a corrected geometric mean time ( _Tmax ) for the peak plasma concentration of the estrone from about 4 hours to about 8 hours (e.g., from about 4.99 hours to about 7.34 hours). In some embodiments, the administration of a suppository to a patient comprises: 1) administering a therapeutically effective amount of a compound of the invention in a plasma sample from a patient comprising 1) about 10 pg * hr / ml to about 16 pg * hr / ml (e.g., about 10.34 pg * hr / pg * hr / ml) corrected peak-to-peak plasma concentration (C _max ) of estrone sulfate; 2) a corrected curve geometric mean area (AUC) of the estrone sulfate from about 56 pg * hr / mL to about 84 pg * hr / mL (e.g., about 56.61 pg * hr / mL to about 83.25 pg * hr / _0-24 ; And 3) a corrected geometric mean time (T _max ) for the peak plasma concentration of estrone sulfate from about 4 hours to about 7 hours (e.g., from about 4.67 hours to about 6.86 hours) .

In some embodiments, the suppository comprises about 10 [mu] g of estradiol. In some such embodiments, administration of the suppository to the patient results in a plasma sample from the patient having a concentration of about 4.7 pg / ml to about 7.6 pg / ml (e.g., about 4.80 pg * hr / ml to about 7.50pg * hr / ㎖) the corrected geometric mean peak plasma concentration of estradiol in the range (it is possible to provide a C _max). In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in about 2.3 pg * hr / ml to about 3.8 pg * hr / ml of estradiol when evaluated at 14 days in a plasma sample from the patient, (C _max ) of estradiol in the range of about 0.40 pg / ml (e.g., about 2.40 pg * hr / ml to about 3.75 pg * hr / ml)

In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in a corrected geometric mean peak plasma of estradiol of about 6.0 pg / ml when evaluated on a day, Concentration (C _max ). In some embodiments, administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in a corrected geometric mean peak plasma concentration of estradiol of about 3.0 pg / ml when evaluated at 14 days in the plasma sample from the patient C _max ) < / RTI >

In some embodiments, the administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in a plasma sample from the patient having a concentration of about 17.5 pg / ml to about 27.4 pg / ml (e.g., (AUC) _0-24 of estradiol ranging from about 17.52 pg * hr / ml to about 27.37 pg * hr / ml). In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in a dose of about 10.9 pg * hr / ml to about 17.2 pg * hr / ml of estradiol when evaluated at 14 days in a plasma sample from the patient, (AUC) _0-24 of the estradiol in the range of from about 10.96 pg * hr / ml to about 17.13 pg * hr / ml.

In some embodiments, the administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in a corrected curve of estradiol of about 21.9 pg * hr / ml when evaluated on a day in a plasma sample from the patient Geometric mean area (AUC) _0-24 is provided. In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol in the patient results in a corrected curve geometric mean of about 13.7 pg * hr / ml of estradiol when evaluated at 14 days in plasma samples from the patient Area (AUC) _0-24 can be provided.

In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in a plasma sample from the patient having a concentration of about 0.6 pg / ml to about 1.1 pg / ml (e.g., , About 0.64 pg / ml to about 1.0 pg / ml) of estradiol (C _avg ). In some embodiments, the administration of a suppository comprising about 10 [mu] g of estradiol to the patient comprises from about 0.1 pg / ml to about 0.3 pg / ml of estradiol when evaluated at 14 days in a plasma sample from the patient (C _avg ) of estradiol in the range of about 0.16 pg / ml to about 0.25 pg / ml).

In some embodiments, the administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in an average concentration of estradiol (C _avg ) of about 0.8 pg / ml when evaluated on a day in a plasma sample from the patient, Lt; / RTI > In some embodiments, administration of suppositories comprising about 10 μg of estradiol to the patient provides an average concentration of estradiol (C _avg ) of about 0.2 pg / ml when evaluated on a plasma sample from a patient at 14 days can do.

In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in 1) about 72.1 pg / ml to about 112.8 pg / ml (e.g., about 72.16 pg / Ml to about 112.75 pg / ml); a corrected geometric mean peak plasma concentration ( _Cmax ) of the estron conjugate; And 2) an average concentration (C _avg ) of the estron conjugate in the range of about 6.3 pg / ml to about 10.1 pg / ml (e.g., about 6.40 pg / ml to about 10.00 pg / ml) ≪ / RTI >

In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in 1) a corrected geometric mean peak plasma concentration ( _Cmax ) of about 90.2 pg / ml of the estrone conjugate in plasma samples from the patient ); And 2) an average concentration (C _avg ) of the estrone conjugate of about 8.0 pg / ml when assessed on day 1.

In some embodiments, the administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in a plasma sample from the patient having 1) about 110.3 pg * hr / ml to about 172.6 pg * hr / ㎖ (e. g., about 110.40pg * hr / ㎖ to about 172.50pg * hr / ㎖; or about 138pg * hr / ㎖) unregulated curve ₀ arithmetic average area (AUC) in the range of estradiol of the diol _-24 ; 2) Estradiol in the range of about 4.6 pg / ml to about 7.8 pg / ml (e.g., about 4.61 pg / ml to about 7.20 pg / ml; or about 5.76 pg / ml) Corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) from about 87.9 pg * hr / ml to about 137.4 pg * hr / ml (e.g., about 88.00 pg * hr / ml to about 137.50 pg * hr / ml, or about 110 pg * 0.0 > (AUC) < / RTI > of the estradiol in the range of _0-24 ; And 4) an estradiol in the range of about 3.6 pg / ml to about 5.8 pg / ml (e.g., about 3.67 pg / ml to about 5.74 pg / ml; or about 4.59 pg / (C _{avg [0-24]} ). &_Lt; / RTI >

In some embodiments, the administration of suppositories comprising about 10 [mu] g of estradiol to the patient results in a plasma sample from the patient having: 1) about 370.3 pg * hr / ml to about 578.8 pg * hr / ㎖ (e. g., about 370.40pg * hr / ㎖ to about 578.75pg * hr / ㎖; or about 463pg * hr / ㎖) the range of estrone ratio adjustment of the curve _O-arithmetic mean area (AUC) ₂₄ ; 2) Calibration of estrone in the range of about 15.4 pg / ml to about 24.2 pg / ml (e.g., about 15.44 pg / ml to about 24.13 pg / ml; or about 19.3 pg / ml) The arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) from about 371.1 pg * hr / ml to about 580.1 pg * hr / ml (e.g., about 371.20 pg * hr / ml to about 580.00 pg * hr / ml, or about 464 pg * hr 0.0 > (AUC) < / RTI >_0-24; And 4) a concentration of estrone in the range of about 15.4 pg / ml to about 24.2 pg / ml (e.g., about 15.44 pg / ml to about 24.13 pg / ml; or about 19.3 pg / ml) And one or more parameters selected from a calibrated arithmetic mean peak plasma concentration (C _{avg [0-24]} ).

In some embodiments, administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in a plasma sample from the patient comprising 1) about 4,745.5 pg * hr / ml to about 7,414.9 pg * (AUC) of the estrone conjugate in the range of about 1 mg / hr / ml (e.g., about 4,745.60 pg * hr / ml to about 7,415.00 pg * hr / ml; or about 5,932 pg * hr / _0-24 ; 2) Calibration of the estron conjugate in the range of about 197.5 pg / ml to about 308.8 pg / ml (e.g., about 197.60 pg / ml to about 308.75 pg / ml; or about 247 pg / ml) The arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) when assessed at 14 days, about 7,182.3 pg * hr / ml to about 11,222.6 pg * hr / ml (e.g., about 7,182.40 pg * hr / ml to about 11,222.50 pg * hr / hr / ml) of the estron conjugate in the range of _0-24 ; And 4) an estrogen conjugate in the range of about 299.1 pg / ml to about 467.6 pg / ml (e.g., about 299.20 pg / ml to about 467.50 pg / ml; or about 374 pg / ml) And one or more parameters selected from a calibrated arithmetic mean peak plasma concentration (C _{avg [0-24]} ).

In some embodiments, the suppository may comprise about 4 g of estradiol. In this case, the administration of the suppository to the patient comprises: 1) a corrected geometric mean peak plasma concentration (C _max ) of estradiol from about 4 pg * hr / ml to about 8 pg * hr / ml; 2) a corrected curve geometric mean area (AUC) of estradiol from about 16 pg * hr / ml to about 26 pg * hr / ml _0-24 ; And 3) a corrected geometric mean time (T _max ) for the peak plasma concentration of estradiol of about 0.25 hours to about 2 hours. In some embodiments, the administration of the suppository to the patient comprises: 1) a corrected geometric mean peak plasma concentration (C _max ) of the estrone from about 1 pg * hr / ml to about 3 pg * hr / ml; 2) corrected geometric mean area (AUC) of the estrone from about 8 pg * hr / ml to about 13 pg * hr / ml _0-24 ; And 3) a corrected geometric mean time (T _max ) for a peak plasma concentration of estrone of from about 1 hour to about 4 hours. In some embodiments, the administration of the suppository to the patient comprises: 1) determining a corrected geometric mean peak plasma concentration (C _max ) of the estrone sulfate from about 4 pg * hr / ml to about 7 pg * hr / ; 2) a corrected curve geometric mean area (AUC) of the estrone sulfate from about 22 pg * hr / ml to about 34 pg * hr / ml _0-24 ; And 3) a corrected geometric mean time (T _max ) for a peak plasma concentration of estrone sulfate from about 1 hour to about 3 hours.

In some embodiments, the suppository comprises about 4 ug of estradiol. In some such embodiments, the administration of the suppository to the patient comprises: 1) administering to the patient a plasma sample comprising: 1) about 2.0 pg / ml to about 3.3 pg / ml (e.g., about 2.08 pg * hr / * hr / ml) of the estradiol in the range of the corrected geometric mean peak plasma concentration (C _max ) of estradiol; And 2) a range of espresso in the range of about 9.5 pg * hr / ml to about 15.1 pg * hr / ml (e.g., about 9.60 pg * hr / ml to about 15.0 pg * hr / And one or more parameters selected from the calibrated geometric mean area (AUC) _0-24 of the diol. In some embodiments, administration of a suppository comprising about 4 μg of estradiol to the patient results in 1) from about 1.0 pg * hr / ml to about 1.7 pg * hr / ml of estradiol in a plasma sample from the patient A corrected geometric mean peak plasma concentration (C _max ) of estradiol in the range of about 5.4 pg * hr / ml to about 1.63 pg * hr / (AUC) _0-24 of estradiol in the range of 9.1 pg * hr / ml (e.g., about 5.76 pg * hr / ml to about 9.0 pg * hr / You can provide parameters.

In some embodiments, administration of a suppository comprising about 4 μg of estradiol to the patient results in 1) a corrected geometric mean peak plasma concentration of estradiol of about 2.6 pg / ml (C _max ); And 2) a corrected curve geometric mean area (AUC) _0-24 of estradiol of about 12 pg * hr / ml when evaluated on day 1. In some embodiments, administration of a suppository comprising about 10 [mu] g of estradiol to the patient results in 1) a corrected geometric mean peak plasma concentration of estradiol of about 1.3 pg / ml (C _max ); 2) When evaluated at 14 days, one can provide one or more parameters selected from a corrected curve geometric mean area (AUC) _0-24 of estradiol of about 7.2 pg * hr / ml.

In some embodiments, the administration of suppositories comprising about 4 [mu] g of estradiol to the patient is between about 0.3 pg / ml to about 0.5 pg / ml (e.g., and it provides from about 0.32pg / ㎖ to about 0.5pg / ㎖) the corrected geometric mean peak plasma concentration (C _max) of estrone conjugate ranging.

In some embodiments, the administration of suppositories comprising about 4 [mu] g of estradiol in the patient results in a corrected peak geometric mean plasma plasma concentration of about 0.4 pg / ml of the estrone conjugate, Concentration (C _max ).

In some embodiments, the administration of a suppository comprising about 4 [mu] g of estradiol to the patient results in a plasma sample from the patient having: 1) about 73.3 pg * hr / ml to about 114.7 pg * (AUC) of the estradiol in the range of about 1 mg / hr / ml (e.g., about 73.36 pg * hr / ml to about 114.63 pg * hr / ml; or about 91.7 pg * hr / _0-24 ; 2) of estradiol in the range of about 3.1 pg / ml to about 4.8 pg / ml (e.g., about 3.14 pg / ml to about 4.90 pg / ml; or about 3.92 pg / ml) Corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) from about 69.7 pg * hr / ml to about 108.9 pg * hr / ml (e.g., about 69.76 pg * hr / ml to about 109.00 pg * hr / ml, or about 87.2 pg * hr / ml) of unmodified curved arithmetic mean area (AUC) of the estradiol _0-24 ; And 4) an amount of estradiol in the range of about 2.8 pg / ml to about 4.6 pg / ml (e.g., about 2.90 pg / ml to about 4.54 pg / ml; or about 3.63 pg / (C _{avg [0-24]} ). &_Lt; / RTI >

In some embodiments, the administration of a suppository comprising about 4 μg of estradiol to the patient results in 1) at least one of the following: 1) about 231.9 pg * hr / ml to about 362.4 pg * hr / ㎖ (e. g., about 232.00pg * hr / ㎖ to about 362.50pg * hr / ㎖; or about 290pg * hr / ㎖) the range of estrone ratio adjustment of the curve _O-arithmetic mean area (AUC) ₂₄ ; 2) calibrated estrone in the range of about 10.3 pg / ml to about 16.3 pg / ml (e.g., about 10.40 pg / ml to about 16.25 pg / ml; or about 13 pg / ml) Arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) from about 261.5 pg * hr / ml to about 408.8 pg * hr / ml (e.g., about 261.60 pg * hr / ml to about 408.75 pg * hr / ml, or about 327 pg * hr 0.0 > (AUC) < / RTI >_0-24; And 4) a concentration of estrone in the range of about 10.8 pg / ml to about 17.1 pg / ml (e.g., about 10.88 pg / ml to about 17.00 pg / ml; or about 13.6 pg / And one or more parameters selected from a calibrated arithmetic mean peak plasma concentration (C _{avg [0-24]} ).

In some embodiments, administration of a suppository comprising about 4 μg of estradiol to the patient results in 1) in a plasma sample from the patient: 1) about 4,062.3 pg * hr / ml to about 6,347.6 pg * (AUC) of the estronic conjugate in the range of about 1 mg / hr / ml (e.g., about 4,062.40 pg * hr / ml to about 6,347.50 pg * hr / ml; or about 5,078 pg * hr / _0-24 ; 2) Calibration of the estron conjugate in the range of about 172.7 pg / ml to about 270.1 pg / ml (e.g., about 172.80 pg / ml to about 270.00 pg / ml; or about 216 pg / The arithmetic mean peak plasma concentration (C _{avg [0-24]} ); 3) from about 4,138.3 pg * hr / ml to about 6,466.3 pg * hr / ml (e.g., about 4,138.40 pg * hr / ml to about 6,466.25 pg * hr / ml, or about 5173 pg * hr 0.0 > (AUC) < / RTI >_0-24; And 4) an estrogen conjugate in the range of about 172.7 pg / ml to about 270.1 pg / ml (e.g., about 172.80 pg / ml to about 270.00 pg / ml; or about 216 pg / ml) And one or more parameters selected from a calibrated arithmetic mean peak plasma concentration (C _{avg [0-24]} ).

The pharmaceutical compositions provided herein are capable of generating a substantially local delivery of estradiol. For example, plasma concentrations of estradiol, estrone, and estrone sulfate measured in a patient's plasma after administration of a pharmaceutical composition as provided herein may be measured after administration of a placebo preparation (i. E., A similar formulation lacking estradiol) And the like. Thus, in some embodiments, plasma concentrations of estradiol, estrone, or estrone sulfate measured after administration of the pharmaceutical compositions provided herein may be lowered by binding to the RLD agent.

In some embodiments, the suppository may comprise about 1 [mu] g to about 25 [mu] g of estradiol. Upon administration of the suppository to the patient, plasma samples from the patient may provide a corrected geometric mean peak plasma concentration ( _Cmax ) of estradiol of less than about 30 pg * hr / ml. For example, administration of a suppository to a patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of estradiol of less than about 18 pg * hr / ml. In some embodiments, the administration of the suppository to the patient provides a corrected curve geometric mean area (AUC) _0-24 of estradiol of less than about 112 pg * hr / ml. For example, administration of a suppository to a patient provides a corrected curve geometric mean area (AUC) _0-24 of estradiol of less than about 63 pg * hr / ml.

In some embodiments, administration of the suppository to the patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of the estrone of less than about 14 pg * hr / ml. For example, administration of a suppository to a patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of the estrone of less than about 7 pg * hr / ml. In some embodiments, administration of the suppository to the patient provides a corrected curve geometric mean area (AUC) _0-24 of the estrone of less than about 65 pg * hr / ml. For example, administration of a suppository to a patient provides a corrected curve geometric mean area (AUC) _0-24 of estrone of less than about 31 pg * hr / ml.

In some embodiments, administration of the suppository to the patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of estrone sulfate less than about 613 pg * hr / ml. For example, administration of a suppository to a patient provides a corrected geometric mean peak plasma concentration ( _Cmax ) of estrone sulfate less than about 16 pg * hr / ml. In some embodiments, administration of the suppository to the patient provides a corrected curve geometric mean area (AUC) _0-24 of estrone sulfate less than about 5291 pg * hr / ml. For example, administration of a suppository to a patient provides a corrected curve geometric mean area (AUC) _0-24 of estrone sulfate less than about 84 pg * hr / ml.

In a further aspect of the present disclosure, the capsule disintegration can be determined. In some embodiments, delivery vehicle disintegration or absorption (delivery after delivery) occurs on the first day of treatment (e.g., 6 hours after the first treatment) and 15 days (e.g., 1 day after the last treatment on day 14) The presence or absence of the vehicle).

The pharmaceutical composition may be formulated as described herein to provide the desired pharmacokinetic parameters in the subject to which the composition is administered (e.g., a female subject). In some embodiments, the pharmaceutical compositions as described herein produce the desired pharmacokinetic parameters for the estradiol in the subject. In some embodiments, a pharmaceutical composition as described herein produces desirable pharmacokinetic parameters for one or more metabolites of estradiol, e.g., estrone or total estrone, in a subject.

After administration of a composition comprising estradiol to a subject, the concentration and metabolism of estradiol can be measured in a sample (e. G., Blood, serum or plasma sample) from the subject. Estradiol is typically reversibly converted to estrone, and both estradiol and estrone can be converted to the metabolite estriol. In postmenopausal women, a significant proportion of the circulating estrone is present as a sulfate conjugate, particularly estrone sulfate. Thus, estrone can be measured for "estrone" quantities (conjugate, eg, estrone sulfate exclusion) and "total estrone" quantities (including both free and unconjugated estrone and conjugated estrone, such as estrone sulfate).

The pharmaceutical compositions of the present disclosure can be characterized for one or more pharmacokinetic parameters of estradiol or its metabolites after administration of the composition to a subject or a population of subjects. These pharmacokinetic parameters include AUC, C _max , C _avg and T _max . AUC is the determination of the area under the curve (AUC) by plotting the drug, serum or plasma concentration along the ordinate (Y axis) with respect to time along the abscissa (X axis). AUC is a commonly used tool in the widely understood pharmaceutical field and is extensively described. C _max is widely understood in the art as an abbreviation for the maximum drug concentration in the blood, serum or plasma of a subject. T _max is well understood in the art as an acronym for time to reach the maximum drug concentration in the blood, serum or plasma of a subject.

In some embodiments, one or more pharmacokinetic parameters such as AUC, C _max , C _avg, or T _max are measured for estradiol. In some embodiments, one or more pharmacokinetic parameters such as AUC, C _max , C _avg or T _max are measured for estrone. In some embodiments, one or more pharmacokinetic parameters such as AUC, C _max , C _avg, or T _max are measured for the entire estrone. Any pharmacokinetic parameter can be a " calibrated " parameter, and a parameter is measured as a change to a reference value.

Immunochromatography (MS) to determine the levels of estradiol, estrone or total estrone in the sample, high performance liquid chromatography (HPLC) with ultraviolet fluorescence detection, liquid chromatography combined with mass spectrometry (LC-MS) , Tandem mass spectrometry (MS / MS), and liquid chromatography-tandem mass spectrometry (LC-MS / MS). In some embodiments, the values of estradiol, estrone, or total estrone are measured using a verified LC-MS / MS method. Methods for measuring hormone levels are widely described in the literature.

Statistical measurement

In accordance with an embodiment, the pharmacokinetics of the pharmaceutical compositions disclosed herein are determined using statistical analysis. According to an embodiment, the Analysis of Variance ("ANOVA") or the Analysis of CoVariance ("ANCOVA") may be used to determine the amount of a pharmaceutical composition comprising an active pharmaceutical composition (eg, a pharmaceutical composition comprising estradiol (E. G., The same pharmaceutical composition except that no estradiol is present), or a subject receiving treatment with a reference drug. Those skilled in the art will understand how to perform statistical analysis of the collected data.

VIII. Example

The following examples are pharmaceutical compositions, delivery vehicles and combinations thereof. A method for producing the same is also disclosed. Data generated using the pharmaceutical compositions disclosed herein are also disclosed.

Example  1: Pharmaceutical composition

In an embodiment, estradiol is obtained and combined with one or more pharmaceutically acceptable solubilizing agents. Estradiol is usually purchased as a pharmaceutical grade ingredient as undifferentiated estradiol, although other forms may also be used. In an embodiment, the pharmaceutical composition comprises estradiol with a dosage strength of from about 1 [mu] g to about 50 [mu] g. In an embodiment, the pharmaceutical composition comprises 10 μg of estradiol. In an embodiment, the pharmaceutical composition comprises 25 ug of estradiol.

In an embodiment, estradiol is combined with a pharmaceutically acceptable solubilizing agent, and optionally other excipients, to form a pharmaceutical composition. In embodiments, the solubilizing agent is at least one of Capmul MCM, Miglyol 812, Gelucire 39/01, Gelucire 43/01, Gelucire 50/13 and Tefose 63.

Gelucire 39/01 and Gelucire 43/01 each have an HLB value of 1. Gelucire 50/13 has an HLB value of 13. Tefose 63 has an HLB value of 9-10.

Various combinations of pharmaceutically acceptable solubilizers were combined with estradiol and investigated as described in Table 1.

The pharmaceutical composition in Table 1, which is liquid or semi-solid at room temperature, was tested by using a Brookfield viscometer (Brookfield Engineering Laboratories, Md., Marlborough, MA) 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 evaluated at 37 캜 to determine their melting characteristics. The viscosity of the gel may be important during encapsulation of the formulation. For example, in some cases it may be necessary to warm the formulation prior to filling the gelatin capsule. In addition, the melting characteristics of the composition may have a significant effect after administration of the preparation into the body. For example, in some embodiments, the formulation will melt at a temperature below about < RTI ID = 0.0 > 37 C. < / RTI > Pharmaceutical composition 11 (Capmul MCM / Tefose 63) did not melt, for example, at 37 ° C or 41 ° C.

The dispersion evaluation of the pharmaceutical composition shown in Table 1 was carried out. Dispersion evaluation was performed by transferring 300 mg of each vehicle system in 100 mL of 37 DEG C water without agitation and observing the mixing characteristics. The results varied from the formation of oil droplets at the top to the separation of the phases into a homogeneous but turbid dispersion. Generally speaking, it is believed that a formulation that is readily dispersible in an aqueous solution will have better dispersion characteristics upon administration. However, as described below in Examples 7 to 9, when a preparation (for example, formulation 13) in which two phases are formed upon introduction into an aqueous solution instead of being easily dispersed in an aqueous solution (for example, formulation 13) , It has surprisingly been found that it has been found to be the most effective.

Example 2: Delivery vehicle

In an embodiment, the pharmaceutical composition is delivered in a gelatin capsule delivery vehicle. Gelatin capsule delivery vehicles can be prepared, for example, by gelatin, NF (150 Bloom, type B), hydrolyzed collagen (e.g., GELITA GELITA AG (Eberbach, Germany)), glycerin, sorbitol special or other excipients. Sorbitol specials may be obtained commercially and may tend to act as plasticizers and wetting agents.

Various delivery vehicles have been developed, as described in Table 2, Gels A-F. In Table 2, each delivery vehicle A to F has a different ratio of one or more components.

Each delivery vehicle A to F was prepared at a temperature ranging from about 45 < 0 > C to about 85 < 0 > C. Each molten delivery vehicle A to F was cast into film, dried and cut into strips. The strips were cut into uniform pieces weighed about 0.5 g thick to about 0.5 mm. The strips were placed in a USP 2 type dissolution vessel in water or pH 4 buffer solution and the time for complete dissolution was recorded (see Table 2). Delivery vehicle A had the fastest dissolution out of both water and pH 4 buffer solutions.

Example  3: Pharmaceutical compositions and delivery Vehicle

Various combinations of pharmaceutical compositions from Table 1 and Table 2 were prepared. The combinations are listed in Table 3.

An aliquot of each of the pharmaceutical compositions of Table 3 is from about 300 mg to about 310 mg. The batch size is as shown in Table 3. To encapsulate the vehicle system, aliquots of each 300 mg to about 310 mg of the pharmaceutical composition were encapsulated in about 200 mg of the gelatin capsule delivery vehicle. Thus, for example, in Experiment 1, the pharmaceutical composition labeled MCM: 39/01 was encapsulated in gelatin capsule delivery vehicle A for a total encapsulated weight of about 500 mg to about 510 mg. The aliquot size is arbitrary depending on the concentration of estradiol and the desired gelatin capsule delivery vehicle size. One of ordinary skill in the art will readily understand how when the delivery vehicle encapsulates the pharmaceutical composition, how to adjust the amount of estradiol in the pharmaceutical composition to accommodate a given size of delivery vehicle.

Example  4: Estradiol  Solubility

In various experiments, solubilizers were tested to determine if 2 mg of estradiol could be solubilized to 100 mg of the total weight of the pharmaceutical composition. The solubilizer was considered to be suitable when the solubility of estradiol in the solubilizer is at least about 20 mg / g. The undifferentiated estradiol was dissolved in various solubilizers until the estradiol was saturated (3 days of equilibrated estradiol / solubilizer), the undissolved estradiol was filtered off, and the drug produced against the concentration of estradiol by HPLC The initial solubility was determined by analyzing the composition.

Example  5: Pharmaceutical composition

The following pharmaceutical compositions are contemplated.

In the following examples, TX-004HR is a pharmaceutical composition 4, 5 and 6 (4 μg of TX-004HR, 10 μg of TX-004HR and 25 μg of TX-004HR) compared to pharmaceutical composition 7.

Example  6: Process

Figure 1 illustrates an embodiment of a method of making a pharmaceutical composition comprising an estradiol solubilized in an encapsulated Capmul MCM / Gelucire solubilizer in a soft gelatin delivery vehicle 100. In operation 102, the CapmulMCM is heated to 40 ° C ± 5 ° C. Heating can be achieved through any suitable means. Heating may be performed in any suitable vessel, such as a stainless steel vessel. Other pharmaceutical compositions may be prepared using the same general method by substituting various excipients, including solubilizing agents.

In Operation 104, Gelucire was mixed with CapmulMCM to form the final solubilizer. As used herein, any form of Gelucire may be used in operation 104. For example, in operation 104, one or more of Gelucire 39/01, Gelucire 43/01, and Gelucire 50/13 may be used. Mixing is carried out, for example, by impellers, agitators, stirrers or other similar devices used to mix pharmaceutical compositions, as is known to those skilled in the art. Operation 104 can be performed under an inert or relatively inert gas atmosphere, such as a nitrogen gas. Mixing may be carried out in any vessel known to those skilled in the art, such as a stainless steel vessel or steel tank.

In operation 106, the estradiol is mixed with the solubilizing agent. In an embodiment, estradiol is micronized when mixed with a solubilizing agent. In another embodiment, the added estradiol is in a non-undifferentiated form. Impellers, agitators, agitators, or other similar devices used to mix pharmaceutical compositions. Operation 106 can be performed under an inert or relatively inert gas atmosphere, such as a nitrogen gas.

However, in embodiments, the addition of estradiol may be performed prior to operation 104. In this regard, operations 104 and 106 may be interoperable with respect to timing or concurrently with each other.

At operation 110, a gelatin delivery vehicle is prepared. Any of the gelatin delivery vehicles described herein may be used in operation 110. In embodiments, gelatin, hydrolyzed collagen, glycerin, and other excipients are combined at a temperature ranging from about 45 ° C to about 85 ° C and made into a film. Mixing may occur in other vessels used to make steel tanks or gelatin delivery vehicles. Mixing may be facilitated by other devices used to combine the contents of the impeller, agitator, agitator or gelatin delivery vehicle. Operation 110 can be performed under an inert or relatively inert gas atmosphere, such as a nitrogen gas. In an embodiment, the gelatin delivery vehicle mixture is degassed prior to being used to encapsulate the pharmaceutical composition.

In operation 112, the gelatin delivery vehicle encapsulates the pharmaceutical composition according to well known protocols to those skilled in the art. In operation 112, a soft gelatine capsule delivery vehicle is prepared by combining the pharmaceutical composition prepared in operation 106 with the gelatin delivery vehicle prepared in procedure 110. Gelatin may be encircled around the material so that it may be partially or fully encapsulated, or gelatin may also be injected or otherwise filled with the pharmaceutical composition prepared in procedure 106.

In an embodiment, operation 112 is completed in a suitable die to provide the desired shape. Vaginal soft gel capsules can be prepared in a variety of geometries. For example, vaginal soft gel capsules may be shaped into teardrops, cones with frusto-conical ends, cylinders, cylinders with larger " cap " parts as illustrated in FIG. 2, or other shapes suitable for insertion into vagina. The resulting pharmaceutical composition encapsulated in a soft gelatin delivery vehicle can be inserted digitally or by an applicator.

Example  7: vulva-vaginal atrophy VVA ) On the improvement of Estradiol  Study of pharmaceutical composition

The aim of this study was to evaluate the efficacy and safety of a pharmaceutical composition comprising 10 μg of estradiol (ie, pharmaceutical composition 2), to treat normal to severe symptoms of VVA associated with menopause 14 days after treatment , And the variability of the voiceless vaginal atrophy end point. In addition, systemic exposures to estradiol from single and multiple doses of pharmaceutical compositions were investigated.

This study was designed to evaluate the safety and efficacy of pharmaceutical compositions to reduce the normal to severe symptoms of vaginal atrophy associated with menopause and to investigate systemic exposure to estradiol after vaginal administration once daily for 14 days of the pharmaceutical composition Randomized, double-blind, placebo-controlled trial to assess the efficacy of a single-phase,

Postmenopausal subjects who met study participation criteria were randomized to one of two treatment groups (pharmaceutical composition or placebo). During the screening period, subjects were asked to self-assess the symptoms of VVA, including vaginal dryness, vaginal or vulvar irritation or itching, dysuria, vaginal pain associated with sexual activity, and vaginal bleeding associated with sexual activity. Subjects with moderate to severe symptoms of at least one self-assessed VVA identified by the subject as the most annoying were eligible to participate in the study.

Clinical evaluation was performed at this point:

Screening period (28 days or less);

Visit 1 - Randomization / baseline (1 day);

Visit 2 - Middle (8 days); And

Visit 3 - End of treatment (15 days).

Eligible subjects are randomized in a 1: 1 ratio to receive a pharmaceutical composition comprising 10 μg of estradiol or an equivalent placebo soft gel capsule, and the first dose of the study drug is administered self-administered Respectively. Serial blood samples for monitoring estradiol levels were collected at 0.0, 1.0, 3.0, and 6.0 hours on day 1 for the first dose administration. The subject remained at the clinical site until completion of the 6 hour blood collection and returned to the clinical facility for additional single blood collection for the measurement of estradiol concentration on the 8th day (pre-morning dose) and on the 15th day. The subjects were instructed to self-administer the research medication assigned to the vagina once daily at approximately the same time (± 1 hour) each morning, provided sufficient study medication was available until the next scheduled visit. Each subject is provided with a diary, where the subject was asked to record daily the medication administration date and time being investigated. The subjects returned to the clinical facility on the 8th day for the interim visit and on the 15th day for the end of the treatment assessment and after the study check. On day 1 (6 hours after dosing) and 15 days, the investigator evaluated the capsule disintegration state.

The study involved a screening period of up to 28 days prior to randomization and a 14 day treatment period. The choice of dosage strength (10 μg of estradiol) and treatment regimen (once daily for 2 weeks) was based on FDA findings on the safety and efficacy of RLD.

(Planned and analyzed) Object  Number

A postmenopausal female subject of no more than 50 (25 per treatment group) aged 40 to 75 years with normal to severe VVA symptoms was randomized. Fifty subjects were enrolled, 48 subjects completed the study, and 48 subjects were analyzed.

Key criteria for diagnosis and inclusion

Fifty female subjects were enrolled in the study. Postmenopausal women aged 40 to 75 years with an average age of 62.3 were enrolled. The average weight (kg) of the subject was 71.2 kg in the range of 44.5 to 100 kg. The average height (cm) of the object was 162.6 cm in the range of 149.9 to 175.2 cm and the BMI (kg / m 2) was 26.8 kg / m 2 in the range of 19 to 33 kg / m 2. The criteria for inclusion in the study are the most annoying and include the normal to severe symptoms of at least one VVA identified by the subject, such as vaginal dryness, dyspareunia, vaginal or vulvar irritation, tingling or itching, dysuria, Self-check of bleeding; Up to 5% epidermal cells in vaginal smear cytology; A vaginal pH above 5.0; And an estradiol level of 50 pg / ml or less. Preliminary studies Based on physical examination, clinical laboratory testing, pelvic exam and mammography, subjects who were otherwise considered as generally healthy were enrolled.

Estradiol  10 [mu] g or placebo, dose, and mode of administration

Subjects were randomly assigned to self-administration (with 1: 1 assignment) of one of the following treatments once daily for 14 days:

Treatment A: The pharmaceutical composition of Example 5 (pharmaceutical composition 2: 10 g of estradiol); or

Treatment B: A placebo soft gel capsule containing the same formulation as Treatment A, except for 10 μg of estradiol.

The estradiol preparation was a bright pink soft gel capsule in the shape of a tear drops. Treatment B had the same composition, appearance, and route of administration as Treatment A, but did not contain estradiol.

Period of treatment

The study involved a screening period of up to 28 days prior to randomization and a 14 day treatment period.

Criteria for Evaluation

Effectiveness Endpoint:

Changes in maturity index (percentage of vaginal vaginal cells, epidermal vaginal cells, and intermediate vaginal cells) of vaginal smear to 15 days from baseline (screening). The data for this end point is shown in Tables 6 to 8.

Change in vaginal pH from baseline (screening) to 15 days. The data for this end point is shown in Table 9.

Changes in the severity of the most annoying symptoms from baseline (randomization) to 15 days: (1) vaginal dryness; (2) vaginal or vulvar irritation, tingling or itching; (3) difficulty in urination; (4) dyspepsia; (5) vaginal bleeding associated with sexual activity. The data for this end point is shown in Table 13 and Table 15. < tb > < TABLE >

Changes in the evaluation of the investigator of the vaginal mucosa from baseline (randomization) to 15 days. The data for this end point is shown in Tables 18 to 21. < tb > < TABLE >

Unless otherwise indicated, efficacy endpoints were recorded as a change from visit 1 - randomization / baseline (1 day) to visit 3 - end of treatment (15 days), except for vaginal bleeding, which is indicated as a treatment success or failure.

Safety As part of the endpoint, the following were evaluated:

Vitality signs, weight, changes in physical examination, pelvic and breast examinations, and side effects.

The concentration of estradiol at each sampling time.

Peak concentration of estradiol at 1 day and at the peak of the sampling time.

To determine the amount of residual delivery vehicle, the delivery vehicle disintegration remains in the vagina after treatment.

The results from the evaluation of the plasma concentration of estradiol are presented in Table 5.

Maturity index result

Screening and Visit 3 - Vaginal cytology data were collected as vaginal smears from the vaginal sidewalls following standard procedures to assess vaginal cytology at the end of treatment (15 days). The change in matured index was assessed as the change in cell composition measured at visit 1 - baseline (day 1) compared to cell composition measured at visit 3 - end of treatment (day 15). Changes in the percentage of epidermal, basolateral and mesial cells from vaginal epithelium from vaginal smears were recorded. The results from this evaluation are presented in Tables 6, 7 and 8.

Changes in pH results

Screening and visit 3 - The pH of the vagina was measured at the end of treatment (15 days). A pH measurement was obtained by pressing the pH indicator strip against the vaginal wall. Subjects participating in the study need to have a pH value of greater than 5.0 at screening. The pH value was recorded in a case report form of the subject. Subjects were recommended not to use douching without sexual activity within 24 hours before measurement. The results from this evaluation are presented in Table 9.

Most annoying symptom data

Subjects were asked to specify symptoms identified as "the most annoying symptoms". During the screening period, all subjects had symptoms of VVA: (1) vaginal dryness; (2) vaginal or vulvar irritation, tingling or itching; (3) difficulty in urination; (4) dyspepsia; (5) Questionnaires were provided to self-assess vaginal bleeding associated with sexual activity. Except for vaginal bleeding associated with sexual activity, each symptom was measured on a scale of 0 to 3, where 0 = no, 1 = mild, 2 = moderate and 3 = severe. Vaginal bleeding associated with sexual activity was measured on a binary scale: N = no bleeding; Y = bleeding. The responses of the subjects were recorded. All randomized subjects were also provided with a questionnaire to self-assess the symptoms of VVA at visit 1 - randomization / baseline (day 1) and visit 3 - end of treatment (day 15). Subjects recorded a self-assessment on a daily diary and collected their answers on the 8th and 15th (end of treatment). Pre-dose assessment results obtained at 1 o'clock visit are considered as baseline data for statistical analysis. Data from this evaluation are presented in Tables 10 and 11.

In general, changes in the most annoying symptoms from baseline were scored according to the evaluation of the VVA symptoms described above. Tables 13 and 14 generally show a comparison between the pharmaceutical composition 1 and the placebo for the most troublesome symptoms and vaginal atrophy symptoms. It is worth noting that this measure, although not statistically significant, has demonstrated a tendency to improve on the 15th day.

With regard to the most annoying symptom data presented in Tables 13 and 14, the time period over which the data was measured is generally considered insufficient to make meaningful conclusions. However, the trend observed as part of this study suggests that the data will show improvement of the most annoying symptoms when data is collected over a longer period of time.

The absence or presence of vaginal bleeding associated with any sexual activity was also measured as one of the most annoying symptoms. Data on vaginal bleeding associated with sexual activity are reported in Table 15.

Estradiol  shame/ Pharmacokinetics  data

In this study, systemic exposure to estradiol was investigated after vaginal administration once daily for 10 days of estradiol for 14 days. The descriptive statistics and observed _Cmax and _Tmax values of plasma estradiol concentrations taken at each sampling time are reported in Table 16 and Table 17. < tb >< TABLE > No statistically significant difference in the systemic concentration of 10 μg of estradiol in the placebo group was observed, suggesting that estradiol is not transported into the bloodstream, and in the bloodstream this would have systemic effects. Rather, it remains in the localized tissue, and the effect of estradiol is thus thought to be localized to the site (i.e., quality) of the administration. The lower limit of detection of the assays used to measure pharmacokinetic data may affect the measured accuracy of the presented PK values. Additional PK studies were performed by more accurate assays in Examples 8 and 9.

For the purpose of monitoring estradiol levels during the study, at doses of 0.0, 1.0, 3.0, and 6.0 hours for doses per day; Before dosing on day 8; And 15 days prior to dosing. Efforts were made to collect blood samples at scheduled times. Sample collection and handling procedures were performed to measure estradiol blood levels according to procedures approved by sponsors and key investigators. All baseline and post-treatment plasma concentrations of estradiol were determined using a validated bioassay (UPLC-MS / MS) method. This data is shown in Table 16 and Table 17. < tb > < TABLE >

Evaluation of vaginal mucosa data

Investigators ranked the vaginal mucosal appearance on day 1 (before dosing) and on day 15. Mild, moderate or severe, where 0 = no, 1 = mild, 2 = mild, 2 = mild, 2 = mild, 2 = mild, = Medium and 3 = Severe. The results from this investigator ranking evaluation are presented in Tables 18, 19, 20 and 21.

relay Vehicle Disintegration  data

Evaluation of capsule disintegration in vagina (presence or absence) at 1 day (6 hours after dosing) and 15 days. The results of this evaluation are shown in Table 22.

Serum hormone level data were collected to determine the serum concentration of estradiol. This data was used for screening inclusion and was determined using standard clinical chemistry methods.

Appropriate for measurement

The choice of efficacy measures used in this study was based on the use of the FDA for the study of estrogen and estrogen / progestin drug products for the treatment of moderate to severe vasomotor symptoms associated with menopause and normal to severe symptoms of vulvar and vaginal atrophy associated with menopause was based on the recommendation (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 reference)).

Standard clinical, laboratory and statistical procedures were used in the experiment. All clinical laboratory procedures were generally accepted and met quality standards.

Statistical methods:

efficacy:

To estimate variability in effect size and efficacy, ANOVA was used to assess changes from baseline differences between subjects receiving 10 μg estradiol and placebo capsules for all efficacy endpoints except vaginal bleeding . In some cases, for example, for some vestibular symptoms, changes from baseline (after a dose response) correlated with baseline values (p < 0.05), baseline values were included as covariates to adjust for this correlation Analysis, ANCOVA). The 90% confidence interval in the difference between 10 μg of estradiol and placebo endpoint measures was determined to evaluate the effect size. The changes from baseline in vaginal bleeding associated with sexual activity were evaluated in terms of the proportion of subjects with treatment success or failure. Any subject reporting hemorrhage at baseline that did not bleed on day 15 was thought to have been successfully treated. Any subject who had bleeding on day 15 was considered a treatment failure, regardless of whether they had the reported baseline bleeding or not. Subjects who did not report bleeding on both baseline and day 15 were classified as unchanged and excluded from the statistical evaluation. Differences in the percentage of subjects having success between the two treatment groups were statistically assessed using Fisher's Exact Test. The results of this difference in ratios are shown in Table 10.

Measurement of compliance

Subjects were asked to complete a diary to record adherence to treatment. The diary was reviewed for adherence to treatment on the 8th and 15th visits. A total of 45 subjects (21 subjects in the 10 μg estradiol group and 24 subjects in the placebo group) were 100% compliant by the treatment regimen.

Due to the research nature of the study, no adjustment was made to the diversity of endpoints.

safety:

The frequency and severity of all adverse events are summarized descriptively by the treatment group.

RESULTS: All subjects (48 subjects) completed the study were included in the first efficacy analysis. The results of the efficacy analyzes are presented in Tables 5, 6 and 7.

conclusion

efficacy

The two week treatment with 10 [mu] g of the pharmaceutical composition resulted in an average decrease in the percentage of stomach cells significantly higher than the placebo treatment, as illustrated in Table 6 (54% vs. 5%, p Lt; 0.0001). At the same time, the mean increase in the percentage of significantly higher epidermal cells was greater for the pharmaceutical composition (35%) than for the placebo capsules (9%) with a very statistically significant difference (p = 0.0002) Lt; / RTI &gt; The difference in pH reduction (0.97 units) between pharmaceutical compositions compared to placebo (0.34 units) was only slightly higher than 0.5 units, but the difference was statistically significant (p = 0.0002) as illustrated in Table 9 ).

The reduction of the severity of the most annoying symptoms is essentially the same for both the pharmaceutical composition and the placebo (about 1 unit), but a reduction in the severity of the individual symptoms of vaginal dryness, irritation and pain during sexual activity is more active than placebo treatment All of the treatments were slightly better. The difference between the two treatments (all less than 0.3 units) was not statistically significant. There was no difference between the two treatments in terms of pain / tingling / tingling (about 0.4 unit decrease) during urination. The length of the study was not long enough to show the separation between the pharmaceutical composition and the most annoying symptoms in the placebo. However, the trend of the most annoying symptoms suggests that by a suitable time period, a significantly significant difference between the two treatments is observed.

Two weeks of treatment with estradiol 10 μg capsules showed no statistically detectable differences with respect to the reduction in severity from baseline according to the evaluator's assessment of vaginal color or vaginal epithelium surface thickness. The pharmaceutical composition capsules showed a statistically significant higher significant effect on vaginal epithelial integrity (-0.34 compared to 0.18, p = 0.0001) and vaginal secretion (-0.64 versus -0.27, p = 0.0401) .

Descriptive statistical analyzes (mean, median, geometric mean, standard deviation, CV, min and max, C _max and T _max ) were used to determine the estradiol concentration at each sampling time, the peak concentration at day 1 and the time . The results from this evaluation are shown in Table 16 and Table 17. &lt; tb &gt;&lt; TABLE &gt;

The pharmaceutical composition comprising 10 μg of estradiol surpassed placebo treatment in terms of improvement in maturation index, reduction in vaginal pH, reduction in epithelial integrity and atrophic effects on vaginal secretion. A lack of statistical significance between the two treatments in relation to the most annoying symptoms and severity of dryness, irritation, pain associated with sexual activity, and individual vaginal atrophy of pain / tingling / The number of subjects and the short treatment period. Too few subjects in the study have vaginal bleeding associated with sexual activity to allow any meaningful evaluation of these vestibular symptoms.

Of the 48 subjects enrolled in the study, 45 subjects were 100% compliant with the treatment regimen. Of the remaining three subjects, one was removed from the study for personal reasons, and the other one of the subjects missed one dose each as a side effect.

safety

Although the daily mean plasma estradiol peak concentration for the pharmaceutical composition is somewhat higher for placebo (ratio of geometric mean = 1.21: 21% of test product (10% estradiol)> placebo), no statistically significant difference was determined I did. However, the test method was suspicious and generated suspicious PK data. Additional PK studies were performed in Examples 8 and 9.

There were no serious side effects in the study.

In general, the pharmaceutical composition containing 10 μg of estradiol was very tolerable when administered intravenously in a regimen once a day for 14 days.

Example  8: PK study (25 제 preparation)

A PK study was conducted to compare the 25 [mu] g formulation disclosed herein (pharmaceutical composition 3) to RLD. The results of a PK study on estradiol are summarized in Table 23. The p-values for this data demonstrate statistical significance as shown in Table 24.

As illustrated in Table 23, the baseline adjusted PK data illustrate that the formulations disclosed herein unexpectedly show a 54% reduction in C _max and a 31% decrease in AUC compared to RLD. This result is desirable since estradiol is only intended for topical absorption. This data suggests a decrease in the circulating levels of estradiol compared to RLD. Moreover, it is worth pointing out that the C _max and AUC levels of estradiol are not statistically different from placebo, suggesting that the formulations disclosed herein have negligible systemic effects. As shown in Table 24, there were no significant differences between the test product and the reference product due to sequence and period effects. However, there was a significant difference due to the therapeutic effect on both C _max and AUC.

The pharmacokinetics of the circulating total estrone, which is the metabolite of estradiol, is shown in Table 25. This data shows that total circulating estrone for the formulations disclosed herein produces a 55% reduction in _Cmax and a 70% reduction in AUC for circulating estrone.

There was a significant difference between the test product and the reference product due to the treatment effect, while there was no significant difference due to sequence and duration effects on C _max . For the AUC, there was a significant difference between the test product and the reference product due to treatment, sequence and duration effects.

The PK for the circulating total estrone sulfate is shown in Table 27. &lt; tb &gt;&lt; TABLE &gt; This data shows that total circulating estrone sulfate for the pharmaceutical compositions disclosed herein produces a 33% reduction in C _max and a 42% reduction in AUC for circulating estrone sulfate.

There was a significant difference between the test product and the reference product due to the therapeutic effect, while there was no significant difference due to the order and duration effects on both C _max and AUC.

Example  9: PK study (10 제 preparation)

A PK study was conducted to compare the 10 [mu] g formulation disclosed herein (pharmaceutical composition 2) relative to RLD. The results of PK studies on estradiol are summarized in Tables 29-40 and Figures 9-14.

PK studies were performed to compare the pharmaceutical compositions disclosed herein with 10 ug of estradiol relative to RLD. The results of PK studies on estradiol are summarized in Tables 29-34, which demonstrate that the pharmaceutical compositions disclosed herein more effectively block the systemic absorption of estradiol. Table 35 shows that the pharmaceutical compositions disclosed herein have a 28% improvement over systemic blood concentration C _max and a 72% AUC improvement over RLD compared to RLD.

PK data for the entire estrone also demonstrated reduced systemic exposure compared to RLD. Table 33 shows that the pharmaceutical compositions disclosed herein reduced systemic exposure by 25% for C _max and by 49% for AUC.

PK data for estrone sulfate similarly demonstrated decreased systemic exposure compared to RLD. Table 37 shows that the pharmaceutical compositions disclosed herein reduced systemic exposure by 25% for C _max and by 42% for AUC.

Example  10: Of VVA  For treatment Estradiol  quality Soft gel  Capsule Randomized , Double-blind, placebo-controlled, multicenter studies.

Under investigation  plan

The study was a randomized, double-blind, placebo-controlled, multicenter study design. Postmenopausal subjects who met study participation criteria were randomly assigned to receive 1: 1: 1: 1, 1: 1, 1: 1, 2, 3, 4 or 5 mg of estradiol soft gel capsule, 10 g of estradiol soft gel capsule, 25 g of estradiol soft gel capsule, Will be randomized. Subjects completed the VVA symptomatic assessment questionnaire and MBS confirmation at 1A on screening visit to determine the eligibility of the study. The subjects were assessed for the presence of vulvar or vaginal atrophy, including vaginal pain associated with sexual activity, vaginal dryness, vulvar or vagular itching or irritation You will be asked to self-assess your symptoms. VVA symptom self-assessment questionnaires, vaginal cytology, vaginal pH and vaginal mucosa will be evaluated at 1B screening visit. This assessment will determine continuing eligibility and will be used as a baseline assessment of the study. Thereafter, the randomized object will complete the questionnaire during visits 3, 4, 5, and 6.

The primary endpoint of efficacy for the study was (A) a change in the percentage of vaginal epidermal cells (from vaginal cytology) to 12 weeks from baseline compared to placebo; (B) a change in the percentage of vaginal (by vaginal cytology) cells from baseline to 12 weeks compared to placebo; (C) changes in vaginal pH from baseline to 12 weeks compared to placebo; And (D) changes in the severity of MBS of dyspareunia (vaginal pain associated with sexual activity) associated with VVA from baseline to 12 weeks compared to placebo.

The secondary efficacy endpoint for the study was (E) the change in percentage of vaginal epidermal cells (by vaginal cytology) from baseline to 2 weeks, 6 weeks, and 8 weeks compared to placebo; (F) changes in the percentage of vaginal (by vaginal cytology) cells from baseline to 2 weeks, 6 weeks and 8 weeks compared to placebo; (G) changes in vaginal pH from baseline to 2 weeks, 6 weeks and 8 weeks compared to placebo; (H) changes in the severity of MBS of dyspareunia (vaginal pain associated with sexual activity) associated with VVA at 2 weeks, 6 weeks, and 8 weeks compared to placebo; (I) changes in vaginal dryness and vaginal or vaginal itching or severity of irritation associated with VVA at 2 weeks, 6 weeks, 8 weeks and 12 weeks compared to placebo; (J) Changes in visual evaluation of vaginal mucosa at 2 weeks, 6 weeks, 8 weeks and 12 weeks from baseline compared to placebo; (K) (as exemplified in the Statistical Analysis Plan (SAP)] Scale visits in subgroups of subjects using the baseline corrected and non-corrected values (1A, 1, Evaluation of standard PK parameters as defined in SAP for serum estradiol, estrone and estron conjugates on days 14 and 84; And (L) a change in female sexual function index (FSFI) at 12 weeks from baseline compared to placebo.

Safety endpoints for the study were (1) side effects; (2) signs of vitality; (3) physical examination findings; (4) discovery of gynecological test; (5) clinical laboratory tests; (6) cervical dilatation test; And (7) endometrial biopsy.

Approximately 100 sites in the United States and Canada to randomize 747 subjects in this study were randomly assigned to each treatment group with a goal of 175 subjects (175 subjects in each active treatment group to complete the 560 subjects, About 175 subjects in the placebo group) (screened for a modified therapeutic intention group, or all subjects taking at least one dose of the pharmaceutical composition disclosed herein). The actual subjects enrolled were 186 subjects in the 4 μg formulation group, 188 subjects in the 10 μg formulation group, 186 subjects in the 25 μg formulation group and 187 subjects in the placebo group for a total of 747 subjects in the study. Within each treatment group, 15 subjects also participated in the PK sub-study. Subjects were assigned to one of four treatment groups: (1) 4 [ mu ] g formulation; (2) 10 [ mu ] g formulation; (3) 25 [ mu ] g formulation; And (4) placebo.

Most subjects participated in the study for 20 to 22 weeks. This is followed by a 6- to 8-week screening period (6 weeks for the unprotected and 8 weeks for the uterus preserved), 12 weeks for the product under investigation and approximately 15 days after the last dose of the product being investigated Observation period was included. The involvement of some subjects lasted up to 30 weeks when an 8-week withdrawal period was needed. Objects abstained from research were not replaced, regardless of the reasons for abstention.

A study schematic diagram is shown in Fig. There were two treatment periods; Vaginal administration once a day in one of the products being investigated for 2 weeks followed by vaginal administration twice a week for 10 weeks.

Object inclusion criteria were: (1) at least 12 months of natural amenorrhea (natural menopause should have a FSH level of more than 40 mIU / ml in women under 55 years of age with a history of hysterectomy without bilateral ovariectomy) ; Or 6 months of natural amenorrhea with a follicle stimulating hormone (FSH) level of greater than 40 ml U / ml; Or postmenopausal female subjects between 40 and 75 years of age (at the time of randomization) by at least 6 weeks after bilateral ovariectomy surgery.

The inclusion criteria were (2) up to 5% epidermal cells in vaginal cytology; (3) a vaginal pH above 5.0; (4) screening visit 1A normal to severe symptoms of vaginal pain associated with sexual activity considered to be the most troublesome vaginal symptom by the subject; (5) screening visit 1B normal to severe symptoms of vaginal pain associated with sexual activity; (6) the occurrence of moderate to severe dyspareunia in the postmenopausal year; (7) the subject is sexually active (ie, has sexual activity due to vaginal penetration within approximately 1 month of screening visit 1A); And (8) subjects who were expected to have sexual activity (due to vaginal penetration) during the performance of the experiment.

For subjects whose uterus was preserved, inclusion criteria also included (9) subjects with acceptable results from evaluable screening endometrial biopsies. At the time of screening, endometrial biopsy reports by two center pathologists defined one of the following: proliferative endometrium; Weak proliferative endometrium; Impaired proliferative pattern; Secretory endometrium; Endometrial tissue, etc. (ie, positive, inactive or atrophic fragments such as endometrial epithelium, line, substrate, etc.); Endometrial tissue insufficient for diagnosis; No confirmed endometrium; No identified tissue; Endometrial hyperplasia; Endometrial malignancy; Or other findings (nonexistent endometriomas, benign endometrial polyps or other endometriomas). At least one pathologist needed to identify enough tissue to evaluate the biopsy.

For subjects with a body mass index (BMI) of 38 kg / m 2 or less, the inclusion criterion is also such that (10) the BMI value is rounded to the nearest integer (for example, 32.4 is rounded to 32 while 26.5 is 27 Rounded).

In general, the inclusion criteria also included (11) by the opinion of the investigator what the subject considered to be in compliance with the protocol and likely to complete the study.

The exclusion criteria are: (1) the use of oral estrogen-, progestin-, androgen-, or SERM-containing drug products within 8 weeks prior to screening visit 1A (withdrawal permitted); Use of percutaneous hormone products within 4 weeks prior to screening visit 1A (allowance for withdrawal is allowed); Use of vaginal hormone products (rings, creams, gels) within 4 weeks prior to screening visit 1A (allowance for withdrawal is allowed); The use of intrauterine progestin within the first 8 weeks of screening visit 1A (allowance for withdrawal is allowed); Use of progestin injectable / injectable or estrogen pellet / injectable within 6 months before screening visit 1A (no entry of abstinence is allowed); Or screening visit 1B Vaginal pH The use of a vaginal lubricant or moisturizer within 7 days prior to assessment.

Exclusion criteria may also include: (2) evidence of a clinically significant medical condition or activity that is detrimental or interfering with an object that includes, for example, hypersensitivity to estrogen; Endometrial hyperplasia; Untreated vaginal bleeding; History of chronic liver or kidney dysfunction / disorder (e.g., hepatitis C or chronic kidney failure); Thrombophlebitis, thrombosis or thromboembolic disorder; Cerebrovascular stroke, stroke or transient ischemic attack; Myocardial infarction or ischemic heart disease; Treatment of malignant tumors or malignant tumors within the past 5 years, excluding basal cell carcinoma of the skin or squamous cell carcinoma of the skin (history of estrogen-dependent tumor formation, breast cancer, melanoma or any gynecologic cancer at any time, Excluded); And endocrine disorders (except regulated hypothyroidism or controlled insulin-independent diabetes mellitus).

Exclusion criteria also include (3) the recent history of known alcohol or substance abuse; (4) history of sexual abuse or spouse abuse likely to interfere with the evaluation of the object of vaginal pain by sexual activity; (5) Current history of the use of large quantities of cigarettes (more than 15 cigarettes per day) or electronic cigarettes; (6) screening visit 1A use of intrauterine devices within 12 weeks; (7) screening visit 1A use of medication under investigation within 60 days; (8) Screening Any clinically significant abnormality in a physical examination, evaluation, electrocardiogram (ECG) or laboratory test; (9) Known pregnancy or positive urine (urine) pregnancy test; And (10) current use of marijuana.

In this study, subjects were not replaced if they were suspended or abstained. At the time of consent, each subject was given a unique object number identifying the clinical site and sequence number. In addition to the assigned object number, the object initials are used for identification. Clinical trials were performed in accordance with standard operating procedures, and regulatory compliance as described by the FDA, the ICH E6 (R1) guidelines, and other relevant control agencies. Clinical trials at clinical sites - compliance with specific audits and database reviews have been achieved.

Statistical method

efficacy. The primary objective of the experiment was to evaluate the effects of estradiol on symptoms of moderate to severe dyspareunia (vaginal pain associated with sexual activity) as vaginal epidermal cells, vaginal vascular cells, vaginal pH and MBS at 12 weeks compared to placebo To evaluate the efficacy of soft gel capsules (4, 10 and 25 [mu] g). Closure procedures were performed to demonstrate multiple comparisons of test placebo and multiple assays of four companion-primary endpoints for each of the three doses of estradiol (4 ug, 10 ug and 25 ug) (Edwards D, Madsen J. " Constructing multiple procedures for partially ordered hypothesis sets. &Quot; Stat Med 2007: 26-5116-24, incorporated herein by reference).

Determine the sample size. To achieve the desired power, the required sample size for each dose versus placebo for each test of the hypothesis in the Modified Therapeutic Intent (MITT) population was calculated using reference data from other studies (Bachman, G., et al . "Effective Treatment of Menopause , 2009. 16 (4): p.719-27; Simon, J., et al. ," Efficacy and safety of low-dose regimens of conjugated estrogens cream administered vaginally " FDA Medical Officer's Review of Vagifem [NDA 20-908, March 25, 1999, Table 6]: Obstetrics & Gynecology, 2008. 112 (5): p. 1053-60; Vaginal Atrophy with an Ultra-Low-Dose Estradiol Vaginal Tablet , p 12., each incorporated herein by reference). Table 41 below provides the effect size, power, and sample size determination for each primary endpoint. In general, subjects in the study met all inclusion / exclusion criteria and had moderate to severe dyspareunia as the most annoying symptom of VVA. Based on power analysis and design considerations, approximately 175 subjects were enrolled per treatment arm.

All subjects with at least one dose of randomly assigned and investigated products formed a Therapeutic Intent (ITT) group. A modified Therapeutic Intent (MITT) population is defined as all ITT subjects with a baseline and at least one follow-up value for each primary endpoint (each subject taking at least one dose of the product under investigation), 1 It was a tea efficacy group. The efficacy-evaluable (EE) population was defined as all MITT subjects who completed the trial, was at least 80% compliant to the product under investigation, had measurements on all primary efficacy endpoints, and had no significant protocol violations It was considered to be protocol compliant. The safety group included all ITT subjects.

Primary efficacy analysis was performed on MITT subjects by support efficacy analysis performed in the EE group. For analytical purposes, the subjects were asked to complete all visits up to and including 6 (12 weeks) of visits considered to have completed the study.

Analysis of efficacy endpoints. For all efficacy endpoints with all numbers including four primary endpoints (mean change from 12 baseline values), the active treatment group mean was compared to placebo using an ANCOVA that adjusted for baseline values.

The baseline and primary and secondary efficacy endpoints were measured at 2 weeks, 6 weeks, 8 weeks, and 12 weeks. The investigated analyzes changed from baseline. Thus, ANCOVA was based on a repeated measures mixed effects model (MMRM) where the random effect was subjective and the two fixed effects were treated group and visit (2 weeks, 6 weeks, 8 weeks and 12 weeks). Baseline measurements and age were used as covariates. ANCOVA was therefore not calculated independently for each study collection period. The analysis was initiated by a complete model, but the duration of interaction (2, 6, 8, and 12 weeks) during treatment visits was only statistically significant (p <0.05).

The following three pairwise comparisons were performed using appropriate ANCOVA controls for 12 week (primary) and 2, 6 and 8 week (second) changes from baseline: (1) active treatment, Placebo; (2) active treatment, medium dose vs. placebo; And (3) active therapy, low dose group versus placebo.

Safety results measurements. Side effects, signs of vital signs, physical examination findings, gynecological test findings, clinical laboratory tests, cervical dilatation and endometrial biopsy were safety parameters. Side effects and SAE are summarized for each treatment group and for all active treatment groups as a percentage of subjects reporting each event. The changes from the actual values and the baseline values of the vital signs and all laboratory test parameters are summarized for each treatment group and for all active treatment groups by descriptive statistics in each evaluation obtained.

Endometrial biopsy evaluation. Three independent pathologists with expertise in treatment and blind gynecologic pathology at the discretion of each other determined the diagnosis of endometrial biopsy slides during the study. Unscheduled endometrial biopsies during all visits 6, early termination and treatment were read by 3 pathologists. Each pathologist's report was categorized into one of three categories: Category 1: hyperplasia / malignant tumor no-proliferative endometrium, weak proliferative endometrium, impaired proliferative pattern, secretory endometrium, endometrial tissue, etc. (Ie, positive, inactive or atrophic fragments such as endometrial epithelium, line, or matrix), endometrial tissue insufficiently diagnosed, no identified endometrium, no confirmed tissue, etc.; Category 2: Hyperplasia - includes hyperplasia with and without atypical cells and complex hyperplasia with or without atypical cells; Category 3: Malignant tumors - Endometrial malignancies.

The final diagnosis was based on the consent of two out of three readings. Consensus reached when two out of three pathologist readers agreed to any of the above categories. For example, any two subcategories of "hyperthyroidism / malignant tumor" were categorized as "Category 1: hyperthyroidism / malignant tumor free." If all three readings are very different (i.e., each is divided into category-category 1, 2 or 3), the final diagnosis was based on the most severe of the three readings.

The analysis group for endometrial hyperplasia was the endometrial hyperplasia (EH) group. At week 12, the EH subjects were randomly assigned (as described in the statistical analysis plan) and took at least one dose of the product being investigated without violation of the exclusion protocol, and had a biopsy during pre-treatment endometrial biopsy and treatment.

Object  cure

The study used a double blind design. The product under investigation was provided as three doses of an estradiol soft gel capsule (4 [mu] g, 10 [mu] g and 25 [mu] g) and as a placebo capsule. All subjects were manually inserting one capsule per day for 14 days (2 weeks) according to one of the following treatment arms, then twice weekly twice a week for 10 weeks:

The product under investigation was distributed to all eligible subjects at Visit 2. Each subject is provided with a total of 30 soft gel capsules of the product under investigation in the labeled bottle, allowing for additional capsules for accidental loss or damage. The second bottle was distributed to visit 5. Each subject was trained by the clinical site to self-administer one capsule per day at approximately the same time for 2 weeks (14 days). The drug administration instructions included "removing the vaginal capsules from the bottle; finding the most comfortable position for you; inserting the capsule with a smaller end-up with a vaginal tube for about 2 inches". Beginning on day 15, each subject received one capsule twice a week for the remaining 10 weeks. The twice-a-week dosing was approximately 3 to 4 days apart and generally did not exceed more than 2 doses in the 7 day period. For example, if a 15-day capacity is inserted on Monday, the next capacity is inserted on Wednesday or Thursday. At randomization visit 2 (day 1), the subject received a first dose of the product being investigated at the clinical facility under the guidance of the research staff.

The investigational estradiol-based soft gel drug product used in the study is a pear shaped, opaque, light pink, soft gel capsule. The capsules contain the solubilized estradiol pharmaceutical composition disclosed herein as Pharmaceutical Compositions 4-7. When the soft gel capsule contacts the vaginal mucosa, the soft gelatin capsule releases the pharmaceutical composition with vaginal fluid. In an embodiment, the soft gelatin capsules are completely soluble.

The placebo used in the study contained excipients in the investigational estradiol-grade soft gel capsules without estradiol (see, for example, pharmaceutical composition 7). The products under investigation and the packaging of the placebo were the same to maintain the investigator's proper blindness. Both the subject and the investigator were not able to confirm the treatment from the packaging or label of the product being investigated.

Subjects were asked to use at least 80% of the products under investigation that were considered to be compliant with the medication being investigated. The number of capsules and diary cards were used to determine compliance with each study visit. The subjects were randomly divided into 4 ug of estradiol soft gel capsule (pharmaceutical composition 4), 10 에 of estradiol soft gel capsule (pharmaceutical composition 5), 25 에 of estradiol soft gel capsule (pharmaceutical composition 6) 1: 1: 1 ratio to receive the pharmaceutical composition 7).

The accompanying medication / treatment was used to treat the chronic or pathologic medical condition at the discretion of the investigator. During the study period the following medications were banned: medications being investigated other than estradiol-grade soft gel capsules under investigation; Estrogen-, progestin-, androgens (i.e., DHEA) or SERM-containing agents other than the product under investigation; Drugs, remedies and supplements known to treat vulvar / vaginal atrophy; Vaginal lubricants and moisturizers (e. G., Replens) are discontinued 7 days before the visit 1B quality pH assessment; And all drugs excluded prior to the study.

Efficacy evaluation

The vaginal cytology smears were collected from the vaginal sidewalls according to standard procedures and sent to the central laboratory to assess vaginal cytology. The percentage of epidermal, ventricular, and mesenchymal cells was determined. All treatment / early termination vaginal cytology results were blinded to sponsors, investigators and subjects.

Screening visits 1B and visits 3, 4, 5 and 6 / The quality of the pH at the end of treatment was determined. Subjects were not allowed to use vaginal lubricants or moisturizers within 7 days of screening pH assessment or at any time after the study. Subjects were advised not to use sexual cleansing without sexual intercourse within 24 hours prior to measurement for all scheduled vaginal pH assessments. After insertion of the unlubricated specimen, the pH indicator strip was applied to the vaginal sidewall until it was wet, being careful to avoid known cervical mucus, blood or semen to affect the vaginal pH. The color of the strip was immediately compared to the colorimetric scale and the measurements were recorded.

During the gynecologic examination, the investigator performed a visual assessment of the vaginal mucosa using a 4-point scale (0 = no, 1 = mild, 2 = moderate and 3 = severe) , Vaginal epithelium surface thickness and quality parameters were evaluated.

The VVA symptomatic assessment questionnaire described below is a self-assessment instrument for subjects with vaginal or vaginal atrophy, including vaginal pain associated with sexual activity, vaginal dryness, vulvar or vaginal itching or irritation. At screening visit 1A, the subject was asked to complete the questionnaire and identify the most annoying symptoms, and the results of the study were used to determine the initial eligibility for the study. At 1A, the subject was also asked to indicate what moderate or severe symptoms were most annoying. Questionnaires were re-administered at 1B of screening visit and were used to determine continuing eligibility for the study.

Randomized subjects were asked to complete assessment questionnaires at visit 3, 4, 5, and 6 o'clock VVA symptom. Subjects were asked to indicate whether they had not experienced sexual activity due to vaginal penetration since the previous visit. The screening visit 1B evaluation results were considered as baseline data for statistical analysis.

The female sexual function index (FSFI) is a simple multidimensional scale for assessing sexual function in females (see Rosen, 2000, supra 26: p. 191-208, incorporated herein by reference). Scale consists of 19 items that assess sexual function over the past four weeks and generate domain scores in six domains: libido, excitement, lubrication, orgasm, satisfaction and pain. Additional validation of the instrument was performed to prolong the validation to include dyspareunia / colon (pain) and multiple sexual dysfunctions (Weigel, M., et al., "The Female Sexual Function Index (FSFI) Cross-Validation and Development of Clinical Cutoff Scores "Journal of Sex & Marital Therapy, 2005. 31:.... p 1-20] reference (incorporated herein by reference)] FSFI was performed at visits 2 and 6 PK Subjects participating in sub-studies were not assessed using the FSFI.

Safety evaluation

Demographic data (age and race / ethnicity) Disclaimer A complete medical history, including scientific, surgical, and psychiatric history and the use of tobacco and alcohol, was recorded at the abstinence / screening visit 1A prior to any withdrawal period, And a review of the current disease and its respective period, and any history of amenorrhea.

A complete physical examination was performed at screening visits 1A and at visit 6 / end of treatment. Physical examination included examination of at least the general appearance of the subject, HEENT (hair, eyes, ears, nose and throat), heart, lung, musculoskeletal, gastrointestinal (GI), nervous system, lymph nodes, abdomen and extremities. The height of the subject was measured only at 1A / 1 visit and the weight (subject lightly dressed) was measured at the abstinence / screening visit 1A and at the end of treatment. BMI was calculated at 1 am visit / screening visit. Vital signs (body temperature, heart rate [HR], respiratory rate [RR], and left ventricular BP [BP]) were measured at all visits after sitting for at least 10 minutes. If the initial BP readings were greater than 140 mmHg systolic or 90 mmHg diastolic, options for a single repeat evaluation performed after 15 minutes were provided. We obtained a standard 12-lead ECG at screening visit 1A and visit 6 or early termination.

Subjects were asked to have a pelvic examination and a cervical dilatation test performed during screening visit 1B and visit 6 or early termination. The uterine cervix was requested for all subjects whose uterus and cervix were not preserved or preserved. For subjects whose uterus was not preserved, a cervical dilatation test was obtained by sampling the apex of the vaginal cuff. All subjects were asked to have a cervical dilatation test performed during the screening, regardless of any recent previous assessments. Subjects who discontinued the study two weeks after the product under investigation were asked to have a cervical screening test. The subjects had a breast scan performed during screening visit 1A and during visit 6 or premature termination.

A licensed gynecologist performed endometrial biopsy at screening and visit 6 / end of treatment. An unscheduled endometrial biopsy was performed during the study when indicated for medical reasons. Initial screening of subjects After the evaluation of the visit indicated that the subject was a different candidate for the study, a screening biopsy was performed at 1B screening visit.

At screening, endometrial biopsy was read by two pathologists. At least one pathologist evaluates endometrial biopsy as endometrial hyperplasia, endometrial cancer, proliferative endometrium, weak proliferative endometrium, or a proliferative pattern with impairment, or at least one pathologist evaluates endometrial hyperplasia with hyperplasia Candidate subjects were excluded from the study when identifying polyps, any degree of linear atypical (eg, atypical nuclei) or cancer. In addition, confirmation of sufficient tissue for biopsy evaluation by at least one pathologist was required for study eligibility. When early endometrial biopsies are performed and both major pathologists report endometrial tissue that is inadequate for diagnosis, the endometrium is not identified, the tissue is not identified, and the subject has until now all other protocol- If the criteria are met, an option for one iteration of the screening endometrial biopsy becomes available. Three pathologists assessed unscheduled biopsy during visit 6 (or early termination) endometrial biopsy and treatment.

Any subject with a diagnosis of endometrial hyperplasia at the time of the study, at the early termination, and at the end of the study, was given a dose of 10 mg of medroxyprogesterone acetate (MPA) for 6 months, . For unscheduled biopsies, histologic diagnosis of endometrial polyps was not forced to abstain unless atypical nuclei were present.

A urine pregnancy test at 1:00 of screening visit was performed unless the subject had a history of tubal ligation, bilateral ovariectomy, or had experienced a menstrual cessation for at least one year, over 55 years of age.

Blood samples for blood chemistry, hematology, coagulation tests and hormone levels, and urine samples for urine analysis were collected and sent to the central laboratory. Blood chemistry (sodium, potassium, chloride, total cholesterol, blood urea nitrogen, iron, albumin, whole protein, aspartate amino converting enzyme (AST), alanine amino converting enzyme (ALT), alkaline phosphatase, creatinine, calcium Glucose, triglycerides (must be at least 8 h fasting), and fasting glucose in excess of 125 mg / dL requiring HgA1C were monitored. Hematology (leukocyte count and classification, red blood cell count, hemoglobin, (Complete blood count (CBC), including hematocrit and platelet counts) were monitored in a subpopulation of the study population. Hormone levels (FSH) (natural amenorrhea or more than 12 months of bilateral oophorectomy) Estradiol, estrone, and estrone conjugates and SHBG were monitored. Urine analysis (appearance, specific gravity, protein and p H).

Pharmacokinetics  evaluation

Seventy-two subjects were also enrolled in the pharmacokinetic (PK) sub-study. In this subject participating in the PK sub-study, time 0h serum blood samples were obtained at screening visit 1A, 1 day and 14 days prior to dosing for baseline. The baseline values were determined by means of two pre-treatment samples. Thereafter, serum blood samples were obtained at the post-dosing time points (2h, 4h, 6h, 10h and 24h) on days 1 and 14. A baseline pretreatment blood sample (time 0h) was collected from each subject prior to the insertion of the product being investigated on study days 1 (visit 2) and 14 (visit 3). Blood samples were taken 2 hours, 4 hours, 6 hours, 10 hours and 24 hours after insertion of the product and after insertion. The last PK sample (approximately 84 days) was obtained 4 days after the last insertion of the product under investigation.

Blood samples were analyzed to determine the area under the curve (AUC), time of maximum concentration (t _max ), minimum concentration (C _min ) and maximum concentration (C _max ). Blood samples were also analyzed to measure levels of estradiol, estrone, and estrone conjugates. The hunger requirement did not apply. Sex hormone binding globulin (SHBG) levels were obtained at the pre-treatment baseline (day 1, visit 2), and on day 14 at 0 h and 84 days of end-hormonal blood collection.

The symptoms / complaints and medication diaries were distributed at all visits, and subjects were instructed to complete. Subjects used the diary to record symptoms / complaints (including discontinued and onset dates and received treatment) and previous medications / treatments (including indication, discontinuation and start date). At each visit, the embryos were copied and redistributed to the subjects. The dosing diary was distributed to visit 2 and visit 3, and the subject was directed to completion. The subjects recorded the product usage and sexual activity under investigation. The dosing diaries distributed at Visit 3 were redistributed to Visits 4 and 5. The duplicates were copied at each visit before redistributing the objects.

Research visit

Study visits were routinely conducted to include the activities listed in Table 43.

Withdrawal periods Visit (if applicable; -14 weeks to 6 weeks). The purpose of this visit is to discuss the study with potential objects and to obtain a prior consent signed and dated before any procedure involving abstinence is carried out. Subjects who agreed to discontinue the current treatment began to withdraw after signing the agreement. The symptoms / complaints and medication diaries were distributed at this visit, and the subject was instructed how to complete the diary. Upon completion of the cancellation period, the object will return to the site for visit 1A.

The activities and assessments performed during the visit are subject to prior consent; Demographic characteristics; Medical / gynecological history; Collection of previous and accompanying drug information; Calculate height, weight, and BMI; Collection of vital signs (body temperature, HR, RR and BP); Distribution of symptoms / complaints diaries and guidance on how to complete the diary.

Screening period visit (visits 1A and 1B). Subjects who did not require abstinence began the screening procedure at 1A at the time of visit as described above for the abstinence period. Except for the vital signs, the procedure performed in the abatement will not be repeated at 1A screening visit. In general, screening visits 1 A and 1 B were completed within 6 weeks (42 days) for subjects without uterus or 8 weeks (56 days) for subjects with uterus. All screening assessments were completed prior to randomization. The investigator reviewed the results from all screening procedures and determined whether the subject was eligible to register for the study

Visit 1A (approximately -6 to 0 weeks). Visit 1A (if applicable) was performed after the period of abandonment or after the subject provided prior consent. Subjects were recommended to be fasting for 8 hours before the visit to collect blood.

Procedures and assessments undertaken at the time of visit are pre-agreed; Demographic characteristics; Medical / gynecological history; Collection of symptoms / complaints and collection of reviews (if applicable) by drug diary and subject from abstinence; A record of previous drug information; Recording and evaluating side effects (AE), starting with the signing of prior agreements; Calculate height, weight, and BMI; Collection of vital signs (body temperature, HR, RR and BP); Physical examination; Breast examination (including mammograms performed up to 9 months before visit 2); Urine pregnancy test as required; Blood and urine sample collection for blood chemistry (at least 8 hours fasting), hematology and urine testing; Serum FSH as required; 12-Lead ECG.

At 1A visit, the VVA symptom self-assessment questionnaire is identified, the most annoying symptom is identified, and the subject is self-identified by MBS as normal or severe pain caused by sexual activity to continue screening. Symptoms / complaints and medication diaries were distributed, and subjects were instructed on how to complete the diary. Subjects were instructed to refrain from using vaginal lubricant for 7 days and sexual intercourse / vaginal cleansing for 24 hours prior to evaluation of vaginal pH performed at Visit 1B.

Visit 1B (approximately -4 to 0 weeks). Visit 1B was performed after the initial screening visit of the subject and after another screening result indicates that the subject is otherwise eligible for the study (preferably around the middle of the screening period).

Procedures and assessments undertaken at the visit include a VVA symptom assessment questionnaire, a subject with normal to severe pain as indicated by sexual activity by vaginal penetration to continue screening; Collection of vital signs (body temperature, HR, RR and BP); Pelvic examination; Investigator evaluation of vaginal mucosa as described above; An evaluation of the vaginal pH (sexual intercourse or vaginal cleansing prohibited within 24 hours before evaluation, and a vaginal pH of the subject over 5.0 to continue screening); Cervical dilatation test; Vaginal cytology (one repetition allowed during screening if no results were obtained from the first smear); Endometrial biopsy performed as described above; Symptoms / complaints by subject and review of drug diary.

Visit 2 (0 weeks; randomization / baseline). Subjects who met participation criteria were randomized to the product being investigated at this visit. Procedures and assessments performed at visits were self-administered FSFI by subjects not participating in the PK sub-study; Review of symptoms / complaints and drug diary by subject; Review of the assessments performed at screening visits and verification of the existence of all inclusion criteria and the absence of all exclusion criteria; Collection of vital signs (body temperature, HR, RR and BP); Randomization by a subject that has been randomized and has met all the entry criteria for which the bottle number has been assigned; Instructions for the distribution of the product being investigated by the subject receiving the first dose of product under investigation under guidance and how to insert the capsule into vaginal if necessary; The distribution of the medication diary and instructions for the completion of the treatment diary, including a record of product usage and sexual activity under investigation.

Visit 3 (2 weeks, 14 days ± 3 days). Procedures and assessments undertaken at the time of visit may include the completion of the VVA symptom assessment questionnaire; Review of symptoms / complaints and drug diary by subject; Collection of vital signs (body temperature, HR, RR and BP); Evaluation of vaginal mucosa; Evaluation of vaginal pH (sexual intercourse or vaginal cleansing prohibited within 24 hours before evaluation); Vaginal cytology; Collection of unused products and bottles being investigated for assessment of compliance / accountability; Redistribution of the product being investigated and, if necessary, repositioning the capsule into the vagina; And a review of the completed dosing diary by the subject.

Visit 4 (6 weeks, 42 days ± 3 days). Procedures and assessments undertaken at the visit may include the completion of the VVA symptom assessment questionnaire; Review of symptoms / complaints and drug diary by subject; Collection of vital signs (body temperature, HR, RR and BP); Evaluation of vaginal mucosa as described above; Vaginal cytology; Evaluation of vaginal pH (sexual intercourse or vaginal cleansing prohibited within 24 hours before evaluation); Collection of unused investigated products for assessment of compliance / accountability; Redistribution of the product being investigated and, if necessary, repositioning the capsule into the vagina; And a review of the completed dosing diary by the subject.

Visit 5 (8 weeks, 56 days ± 3 days). Procedures and assessments undertaken at the visit may include the completion of the VVA symptom assessment questionnaire; Review of symptoms / complaints and drug diary by subject; Collection of vital signs (body temperature, HR, RR and BP); Evaluation of vaginal mucosa as described above; Vaginal cytology; Evaluation of vaginal pH (sexual intercourse or vaginal cleansing prohibited within 24 hours before evaluation); Collection of unused investigated products for assessment of compliance / accountability; Redistribution of the product being investigated and, if necessary, repositioning the capsule into the vagina; And a review of the completed dosing diary by the subject.

Visit 6 (12 weeks, 84 days ± 3 days or early termination). This visit was conducted when the subject abstained from the study prior to visit 6. Procedures carried out at this visit include the completion of the VVA symptom assessment questionnaire; Examination of the subject of medication diary, symptom / complaint, and medication diary by the subject; Collection of blood and collection of urine samples for blood chemistry (at least 8 hours fasting), hematology and urine tests; Vital signs (body temperature, HR, RR and BP) and body weight; Performing 12-lead ECG; Collection of unused products and containers under investigation for assessment of compliance / accountability; Physical examination; Breast examination; Evaluation of vaginal mucosa as described above; An evaluation of the vaginal pH (sexual intercourse or vaginal cleansing prohibited within 24 hours before evaluation, and a vaginal pH of the subject over 5.0 to continue screening); Vaginal cytology; Cervical dilatation test; Endometrial biopsy; Self-administration of FSFI by non-participating subjects in the PK sub-study; Self-administration of a study titled " Tolerability of Product Dose Survey " by the subject.

Follow-up interview ( approximately 15 days after the last dose of the product under investigation ). Each subject receiving the product under investigation received a follow-up phone call approximately 15 days after the last dose of the product under investigation, regardless of the duration of treatment. Follow-up follow-up after all safety evaluations (eg, endometrial biopsy and mammography results) generally occurred. Follow-up should include review of ongoing side effects and any new side effects occurring 15 days after the last dose of the product under investigation; Review of ongoing co-medications and any new co-medications that occurred during the 15 days after the last dose of the product under investigation; And discussion of all endpoints of the study safety assessment and the need for additional follow-up observations or clinic visits.

PK sub-study visit procedure and schedule

Screening visit 1A. In addition to the procedures described above, activities in PK sub-studies may also provide consent to participation in pre-agreement and PK sub-studies by the subject; Included was the collection of serum blood samples during visits to assess baseline values of estradiol, estrone, and estrone conjugates.

Visit 2 (0 weeks, 1 day). In addition to the procedures described above, activity in the PK sub-study also included the collection of serum blood samples obtained prior to administration of the product under investigation (at time 0 h) for the baseline assessment of estradiol, estrone, estrone conjugate and SHBG. After taking a pre-treatment blood sample, the subject self-administered the product under investigation. After administration of the product under investigation, serum blood samples were obtained at 2h, 4h, 6h, 10h, and 24h for estradiol, estrone and estrone conjugates (serum samples generally within ± 5 minutes at 2h and 4h, ± Lt; / RTI &gt; in 15 min and in 10 h and 24 h in 1 h). The subject was released from the site after 10 hours of sampling and was instructed to return to the site the next morning for blood sampling for 24 hours. Subjects were instructed not to self-administer the 2-day dose until instructed to do so by the site staff at the clinical site. The subjects were released from the clinical site after a 24 hour blood sample and a 2 day dose.

Visit 3 (2 weeks, 14 days). The visit must occur on the 14th day without an allowed visit window. In addition to the procedures described above, the PK sub-study included the collection of serum blood samples prior to dosing (at time 0 h) with a 14 day dose for SHBG and PK evaluation. Subjects were self-administered a 14 day dose at the clinical site and serum blood samples were obtained at 2h, 4h, 6h, 10h and 24h for estradiol, estrone and estrone conjugates. The subject was released from the site after 10 hours of sampling and was instructed to return to the site the next morning for blood sampling for 24 hours. Subjects were instructed not to self-administer the 15-day dose until instructed to do so by the site staff at the clinical site. The subjects were released from the clinical site after a 24 hour blood sample and a 15 day dose. Subjects were instructed to administer the next dose of the study drug on the 18th or 19th day and to continue dosing twice a week at the same time of day for each dose.

Visit 6 (12 weeks, 84 days ± 3 days or early termination). Visited at the last IP capacity or 4 days after the early closure. In addition to the procedures described above, serum samples for estradiol, estrone and estrone conjugates and SHBG were taken. PK sub-study visits were routinely conducted to include the activities listed in Table 44.

An adverse effect (AE) in a study is defined as the occurrence of an undesirable medical condition or the deterioration of an already existing medical condition after or during exposure to a pharmaceutical product, whether incidentally associated with the product or not. AE can result from overdosage of the product being investigated. In this study, AE may include undesirable medical conditions that occur at any time, including baseline or withdrawal periods, even though the study therapy is not administered. Relationship to product under investigation

The investigator determined the relationship to the product under investigation for each AE (unrelated, possibly related, or possibly related). The degree of "relevance" of adverse effects on the product under investigation is as follows (non-related-non-time related), other etiology is probably the cause (possible-time association), and other etiology is probably the cause. However, the involvement of the product under investigation can not be ruled out (probable-time associations) and other etiologies are possible, but unexpected. The case may respond to whether the product being investigated is discontinued.

Example  11: Randomized , double Blind , Placebo control, efficacy results of multicenter studies.

Each of the three doses showed statistical significance compared to placebo for the primary endpoint. Each of the three doses showed statistical significance compared to placebo for the secondary endpoint. Table 45 shows the significance of the experimental data for each of the four accompanying-primary endpoints. Each dosage met the respective four co-primary endpoints at a statistically significant level. The 25 mcg dose of TX-004HR demonstrated very statistically significant results at p = 0.0001 level compared to placebo across all four companion-1 endpoints. The 10 mcg dose of TX-004HR demonstrated very statistically significant results at the p = 0.0001 level compared to placebo over all four companion-1 endpoints. The 4 mcg dose of TX-004HR also demonstrated highly statistically significant results at the p = 0.0001 level compared to placebo for endothelial vaginal cells, vaginal vaginal cells, and vaginal pH endpoints, The severity of dyspareunia was at p = 0.0255 level.

A statistical improvement compared to placebo was also observed for all three doses in the first assessment at 2 weeks and lasted for 12 weeks. Pharmacokinetic data for all three doses demonstrate low systemic absorption and support previous phase 1 experimental data. TX-004HR was very tolerant and had no clinically significant differences in side effects compared with women treated with placebo. Serious drug-related side effects were not reported.

As shown in the data below, at 12 weeks in the MITT population (n = 747), all TX-004HR doses significantly reduced the percentage of basal cell and vaginal pH compared to placebo, and the percentage of epidermal cells was significantly And significantly reduced the severity of dyspareunia (p = 0.00001, all except dyspareunia at 4 μg p = 0.0149).

(P <0.00001), the percentage of epidermal cells was significantly increased (p <0.00001), and the severity of dyspareunia was significantly higher at placebo (4 μg p <0.03; 10 μg and 25 μg p <0.02), respectively, by all TX-004HR doses compared to baseline values.

Moderate to severe vaginal dryness was reported at 93% of baseline and significantly improved (p <0.02) for all doses (except 4 μg at 2 weeks) at 2, 6, 8 and 12 weeks. Vulvar and / or vaginal itching or irritation was significantly improved (p <0.05) at 10 μg at 8 weeks and 12 weeks and 25 μg at 12 weeks.

TX-004HR was highly tolerant, had high tolerability, and severe AEs related to therapy were not reported in the safety group (n = 764). There was no clinically significant difference in any AE or treatment-related SAE between TX-004HR and placebo. Very low or negligible whole body levels of estradiol were observed.

All TX-004HR doses were safe and effective, resulting in very low or negligible systemic absorption of E2 in women with VVA and moderate to severe dyspareunia. The occurrence of the effect was seen as fast as two weeks, was maintained throughout the study, and the acceptability was very high. This new product offers promising new treatment options for women who experience menopause VVA.

cytology

Screening and Visit 6 - Vaginal cytology data were collected as vaginal smears from the vaginal sidewalls following the procedure described above to assess vaginal cytology at the end of treatment (84 days). The change in matured index was assessed as the change in cell composition measured at visit 1 - baseline (day 1) compared to cell composition measured at visit 3 - end of treatment (84 days). Changes in the percentage of epidermal, basolateral and mesial cells from vaginal epithelium from vaginal smears were recorded. The results from this evaluation for epidermal cells are presented in Tables 46 and 47, and Figures 10, 11 and 12. The results from this evaluation for basal cells are presented in Tables 48 and 49, and in Figures 13, 14 and 15.

Epidermal phase  cell

This study showed that the formulations disclosed herein over all doses increased the percentage of epidermal cells over all dosages in a statistically significant manner.

Room base  cell

The increase in epidermal cells and the decrease in basolateral cells were statistically significant compared to placebo at week 2 and week 12, including weekly. Administration of the pharmaceutical agent resulted in rapid onset of activity as early as two weeks after the initial administration. Rapid activity can be coupled with rapid absorption that is proven in the pharmacokinetic data presented below.

pH

The vaginal pH was measured at screening and visit 6-treatment end (84 days). pH measurements were obtained as described herein. The results from this evaluation are shown in Tables 50 and 51, and Figs. 16, 17 and 18.

The decrease in vaginal pH was observed at two weeks and at statistically significant levels through the end of the study. Surprisingly, the pH was reduced at all three doses over the full pH unit for all three pharmaceutical formulations tested and all three doses.

The most annoying symptom

Dyspareunia

Subjects were asked to define symptoms identified as "the most annoying symptoms". During the screening period, all subjects were (1) dyspepsia; (2) vaginal dryness; And (3) a questionnaire was provided to self-assess the symptoms of VVA of vaginal or vulvar irritation, tingling or itching. Each symptom was measured on a scale of 0 to 3 where 0 = no, 1 = mild, 2 = moderate and 3 = severe. Each subject was given a questionnaire at each visit and recorded their responses. Questionnaires were also provided to self-assess the symptoms of VVA at each subsequent visit through all randomized subjects at visit 1 and visit 6 - end of treatment (84 days). Subjects recorded daily self-assessments on the diary and collected answers at visits 8 and 15 (end of treatment). Pre-dose assessment results obtained at 1 visit were considered as baseline data for statistical analysis. Data from this assessment of dyspareunia are presented in Table 52 and Table 53. Data from this assessment of dryness are presented in Tables 54 and 55. TABLE 54 &lt; tb &gt;

Each of the 4, 10 and 25 ug formulation tests showed early onset activity at 2 weeks for the most troublesome symptoms of dyspareunia, as evidenced by statistically significant results (as measured by the p-value) . Two weeks later, each dose demonstrated isolation from placebo in the improvement of the most annoying symptoms of dyspareunia.

Combined with the PK data presented below, this result shows that the formulation disclosed herein provides bolus of estradiol within 2 hours of administration, which results in a reduction in severity of dyspareunia as fast as two weeks later. Estradiol was rapidly absorbed in approximately 2 hours, which was significantly earlier than the prior art formulation pursuing an extended release profile. The rapid uptake of estradiol is believed to be the result of administration by liquid preparations.

Surprisingly, 4 [mu] g formulations showed clinical efficacy at 2 weeks with 25 [mu] g and 10 [mu] g dose levels. This data demonstrates that 4 ug is an effective dose and can be effective as early as two weeks after the administration for the most troublesome symptoms of dyspareunia.

Dryness

Each of the 4, 10 and 25 ug formulation tests showed early onset activity at 2 weeks for the most troublesome symptoms of dryness, as evidenced by statistically significant results (as measured by the p-value) . After two weeks, each dose demonstrated the isolation from placebo in the improvement of the most annoying symptoms of dryness.

Irritation / itching

The vulvar and / or vaginal itching or irritation significantly improved (p < 0.05) for 10 [mu] g at 8 and 12 weeks and 25 [mu] g at 12 weeks. Moreover, the trend for 4 μg was an improvement in the itching state compared to a statistically significant state at 12 weeks.

Combined with the PK data set forth below, this result shows that the formulation disclosed herein provides bolus of estradiol within 2 hours of administration, which results in a reduction in the severity of dryness as fast as two weeks later. Estradiol was rapidly absorbed in approximately 2 hours, which was significantly earlier than the prior art formulation pursuing an extended release profile. The rapid uptake of estradiol is believed to be the result of administration by liquid preparations.

Surprisingly, 4 [mu] g formulations showed clinical efficacy at 2 weeks with 25 [mu] g and 10 [mu] g dose levels. This data demonstrates that 4 ug is an effective dose and can be as effective as the two weeks after administration for the most troublesome symptoms of dryness.

As described above, each dose was compared to placebo for changes in the percentage of vaginal epidermal and basolateral cells, vaginal pH, and severity of dyspareunia (co-primary endpoint) from baseline to 12 weeks. (Defined as women with a score of 2 or more after 12 weeks: vaginal epidermal cells in excess of 5%, vaginal pH below 5.0, improvement of one or more categories from baseline dyspareunia scores) compared to placebo in the TX-004HR group Respectively. (Age <56 yr, 57 yr 61 yr and 62 yrs), BMI (24 kg / m2 or less, 25-28 kg / m2 and 29 kg / m2 or more), uterine status, parity and vaginal delivery The analysis of predefined subgroups of companion-primary endpoints was analyzed. Pharmacokinetic (PK) parameters were compared with placebo in a sub-analysis of the main study.

The proportion of responders was significantly higher for all TX-004HR dose groups compared to placebo (p <0.0001 for all). Compared to placebo, all TX-004HR doses significantly improved the percentage of epidermal and dendritic cells, vaginal pH, and severity of dyspareunia at 12 weeks. Subgroup analysis showed generally similar results for epidermal and basal cell percentage and quality of pH regardless of age, BMI, uterine status, parity, and vaginal delivery. The severity of dyspareunia was significantly reduced at 12 weeks by all TX-004HR doses compared to placebo in most subgroups (Table 57A).

The PK sub-analysis (n = 72) described in more detail below found AUC and C _avg parameters for E2 and estrone (E1) by 4 μg and 10 μg of TX-004H similar to placebo. The increase occurred in E2 AUC and C _avg by 25 μg compared to placebo, but was within the normal postmenopausal range. At day 84, E2 levels were similar between the TX-004HR group and placebo, indicating no systemic drug accumulation.

All doses of TX-004HR were associated with robust efficacy, demonstrating statistically significant differences compared to placebo to increase epidermal cells, decrease basal cell and vaginal pH, and reduce the severity of dyspareunia. Age, BMI, uterine status, parity, and vaginal delivery did not generally affect TX-004HR efficacy. This result was caused by negligible systemic absorption of 4, 10, and 25 μg of TX-004HR estradiol dose.

[Table 57A]

At baseline and at 2 weeks, 6 weeks, 8 weeks, and 12 weeks, visual evaluation of the vaginal epithelium, the secondary endpoint of the experiment during gynecological examinations, was performed. Changes in vaginal color, vaginal epithelium integrity, vaginal epithelial surface thickness and vaginal secretion were evaluated using a 4-point scale (0 = no, 1 = mild, 2 = moderate, 3 = severe). Mixed effect model repeatability (MMRM) analysis was used to compare changes from baseline to each time point with placebo.

At baseline, women have an average score of 1.8 for vaginal color, 1.5 for epithelial integrity, 1.9 for epithelial surface thickness, and 1.7 for secretion. This score reflects pale decreased vaginal wall integrity and thickness, and secretion consistent with VVA. Significant improvements from baseline at 2, 6, 8 and 12 weeks (Table 57B; Figures 19a-19d) were observed in comparison with placebo in vaginal hue (white to pink), epithelial integrity, epithelial surface thickness and secretion All three doses of TX-004HR were observed (p <0.001 for all). After 12 weeks, the women in the active TX-004HR treatment group had an average score of less than 1 in all four identified categories. Women's vaginal examination in the three TX-004HR groups reported a higher improvement from baseline in all vaginal parameters examined than placebo subjects at all time points. This improved vaginal visual score was significantly reduced by other observations of the efficacy of TX-004HR (4, 10, and 25 [mu] g) in treating moderate to severe VVA in postmenopausal women by negligible or very low systemic E2 uptake Reflect the measured value.

[Table 57B]

When all subjects were independently analyzed for treatment, there was a statistically significant difference between the total scores of the individual visual inspection scores and severity of dyspareunia (r = 0.31; p <0.0001) and the severity of vaginal dryness (r = 0.38; p <0.0001) Direct correlation was observed. 20A and 20B. Interestingly, women treated with placebo also show some improvement in scores at 2 weeks, while women treated with TX-004HR show continuous improvement over 12 weeks of treatment, while such continuous improvement is similar to placebo Respectively. Three possible explanations for the improvements observed by placebo are the possible lubrication effects of the excipient Miglyol, which is a fractionated coconut oil contained in all soft gel capsules, as they can be expected to improve, vaginal lubrication caused by an increase in sexual activity And / or biases in a portion of the physician performing the examination. Nevertheless, TX-004HR still significantly improved the evaluated signs and symptoms of VVA over placebo.

This tool helps healthcare professionals diagnose VVA and assess their treatment, as the vaginal visual examination by the 4-point instrument correlates positively with dyspareunia and vaginal dryness in this study, And may provide a mediator to initiate discussion with the patient. Several large studies have shown that it is difficult for patients to openly discuss voicing-quality health with a healthcare professional, because the patient believes that they are either blurry, are ignorant of VVA and its treatment, or are not appropriate for discussion. Thus, among 50% of postmenopausal women with symptoms of VVA, far fewer women seek treatment. Vision screening helps doctors identify women at risk for dyspareunia and vaginal dryness and can proactively allow women to participate in conversations about VVA symptoms, such as dyspareunia and dryness, and discuss available treatment options .

Example  12: Randomized , double Blind , Placebo control, multicenter studies Pharmacokinetics  result.

Although some licensed topical estrogens effectively treat VVA, systemic estradiol may increase local dosing. TX-004HR is a new low-dose vaginal soft gel capsule containing solubilized natural estradiol designed to provide excellent efficacy with negligible systemic absorption. Less than three-fold lower systemic estrogen levels were previously reported by TX-004HR compared to licensed low-dose vaginal oestradiol tablets. This study shows that VVA efficacy can be achieved by negligible systemic absorption as measured by PK in postmenopausal women with moderate to severe dyspareunia.

The terms " minimal systemic effect &quot;," low systemic absorption &quot;, and " negligible systemic absorption &quot;, as used herein, means that the disclosed formulations and methods are low in women, especially women with VVA and / or dyspareunia Which results in minimal absorption. Indeed, it has surprisingly been found that the disclosed formulations and methods result in negligible or very low systemic absorption of estradiol in the postmenopausal range. The discovery was substantiated by the examples provided herein and this example demonstrates that the C _max and AUC levels of estradiol for placebo are not statistically variable and this demonstrates that the agents disclosed herein are negligible precursors Effect. Therefore, the disclosed formulations and methods are advantageously used for the treatment of VVA and / or dyspareunia (i.e., the disclosed agents are extremely effective in increasing epidermal cells, decreasing basal cell and decreasing pH) without increasing systemic levels And provides local benefit in the patient having.

PK sub-study was designed to assess the efficacy and safety of TX-004HR (4 μg, 10 μg and 25 μg) in comparison with placebo to treat moderate to severe dyspareunia after menopause, with large, multicentre, double-blind, randomized, Placebo-controlled, phase III trials. Women received TX-004HR or placebo twice a week for two weeks followed by ten weeks for two weeks.

In this study, systemic exposures to estradiol after intravaginal administration of 25 μg, 10 μg, 4 μg, and once daily placebo of estradiol were investigated on days 1, 14, and 84 as described herein. The descriptive statistics of plasma estradiol concentration and observed C _max values taken at each sampling time were recorded for the estradiol, estrone and estron conjugates for all three doses as described in the following table and Figures 21 and 22 Respectively. Serum estradiol, estrone, estrone conjugate and sex hormone binding globulin were measured.

For PK, sera were sampled at 2 hours, 4 hours, 6 hours, 10 hours, and 24 hours before dosing and at 1 day and 14 days for estradiol, estrone (E1) and estrone conjugates (E1C). The adjusted results are listed here; Unadjusted data will be presented. The efficacy endpoint was a change from baseline to 12 weeks for vaginal epithelial cells (%), vaginal basal cells (%), vaginal pH and severity of dyspareunia. The secondary end point was dryness severity and itching / irritation. Blood chemistry was tested at 12 weeks.

The sub-study randomized 72 women (mean age 59 years) from 11 centers. The mean area (AUC) and mean concentration (C _avg ) under the concentration-time curve for estradiol was significantly different from placebo by 4 μg and 10 μg of TX-004HR, but was significantly higher than day 1 (AUC 13.8 h * pg / (5.4 compared to 130 and Cavg 0.4 pg / ml compared to 14.6 days versus 84.6 vs Cavg -0.2 pg / ml compared to 7.1 h * pg / ml for AUC).

The mean estradiol peak concentration (C _max ) was 4 μg (1 day: 2.6 pg / ml; 14 days: 1.3 pg / ml) relative to placebo (1 day: 2.1 pg / (1 day: 6.0 pg / ml; 14 days: 3.0 pg / ml) and very low for 25 μg (1 day: 26.2 pg / ml; 14 Day: 12.0 pg / ml).

E1 and E1Cs AUC, C _avg , C _max and C _min were determined by 25 μg of AUC (2454 compared to 83.0 h * pg / ml), C _max of 10 μg and 25 μg (90.2 And 198.6 pg / ml), compared to placebo, except for E1Cs on day 1 when C _avg was significantly higher by 10 μg (8.0 compared to -33.7 pg / ml).

In the entire study, TX004-HR showed a robust efficacy against symptoms of dyspareunia, vaginal dryness and irritation at 12 weeks by all three doses compared with placebo.

Quality TX-004HR produced negligible or very low systemic absorption in the postmenopausal range of estradiol. TX-004HR improved signs and symptoms of VVA. This study supports the local benefit of estradiol without an increase in systemic exposure.

Pharmacokinetic data for estradiol demonstrate rapid absorption of the formulation disclosed herein for all three doses. Surprisingly, although the pharmacokinetic data are extremely low for all three doses, each dose is extremely effective in increasing epidermal cells, thus reducing epidermal cells and decreasing pH.

The pharmaceutical compositions disclosed herein provide an improved safety profile over other options for treating VVA. The combination of low systemic estradiol was an amazing result for all three doses while retaining efficacy.

Estradiol  density

Estradiol  Area under the curve (0 to 24 hours)

Estradiol C _max

Estradiol C _avg

Estradiol T _max

Estrone concentration

The area under the estron curve (0 to 24 hours)

Estrone C _max

Estrone C _avg

Estrone T _max

Estron conjugate

Area under the estron conjugate curve (0 to 24 hours)

Estron conjugate C _max

Estron conjugate C _avg

Estron conjugate T _max

In the three-phase study, all doses of TX-004HR compared to placebo (MITT n = 747) had 4 companion-1 endpoints from 2 weeks to 12 weeks, and a second endpoint of vaginal dryness at 6 weeks and 12 weeks Significantly improved vaginal and / or vaginal itching or irritation (4 ug excluding, p = 0.0503), and was very idiomatic without reported treatment-related severe AE. Three phase PK studies (n = 72) showed no difference in systemic E2 levels for 4 μg and 10 μg of TX-004HR for placebo, as measured by AUC and C _avg . E2 AUC and C _avg by 25 μg of TX-004HR were higher than placebo, but the mean concentrations were within normal postmenopausal ranges (Table 142). At day 84, E2 levels were similar to placebo and did not show systemic drug accumulation. The SHBG concentration was not changed by treatment. The two biphasic studies (n = 36 for each) of 10 μg and 25 μg of TX-004HR resulted in statistically significantly lower E2 uptake than the licensed E2 tablets at the same dose and 25 μg of TX-004HR Demonstrate a one-third lower AUC than that of the licensed product (Table 143).

As robust efficacy was demonstrated at 2 weeks and 12 weeks early in all three doses, 4 μg and 10 μg of TX-004HR showed negligible systemic E2 uptake while 25 μg showed very low systemic absorption of E2 . 10 [mu] g and 25 [mu] g of TX-004HR showed lower systemic E2 exposure than the equivalent dose of the licensed E2 tablet. The absence of a clinically significant increase in E2 concentration, coupled with data consistent with a lack of systemic effects (eg, no increase in SHBG), suggests that TX-004 HR has excellent efficacy by negligible or very low systemic exposure .

TX-004HR 25 mcg pharmacokinetic (PK) profile was assessed for 4 hours after dosing compared to the effect of being in the angora position for 4 hours after dosing. In two studies, at the same site, the same 16 healthy postmenopausal female subjects were fasting for at least 10 hours prior to dosing and 4 hours after dosing. The subject received a dose of 25 mcg of TX-004HR administered intravaginally by a trained female researcher. After the first dose, the subject was asked to be in a sitting position for 4 hours after dosing. After the second dose, after a 5 minute break, the subjects were refrained from walking and waiting in the clinic for 4 hours after dosing. Blood samples were collected at predetermined intervals up to 24 hours after dosing. Plasma samples were analyzed for estradiol using LC-MS / MS. See FIG. 23, for example. The PK parameters were calculated on the individual and group average criteria with baseline correction.

The mean C _max and AUC _{0 -24} of estradiol were not significantly different by walking than supine. On an individual subject basis, most showed C _max and AUC _{0 - 24} levels by walking similar to supine. There was no indication of a posture that had an effect on water absorption as evidenced by similarity in group mean and individual subject T _max . In addition, there is no difference between the group mean profiles when compared to the individual viewpoint criterion, further demonstrating that the posture has no effect on absorption. TX-004HR The systemic exposure of estradiol at 25 mcg was generally low and occurred independently of walking or amblyopia for 4 hours after dosing. An important advantage of the formulation is that, as opposed to other known agents requiring the subject to be in the pelvic position after administration, the female may be almost immediately following the administration of the formulation. In general, other known formulations should be administered prior to falling asleep at night due to the requirement that the pharmaceutical composition disclosed herein adhere to vaginal tissue, the capsules dissolve rapidly, the formulation is released to vaginal tissue , This requirement is not required in the pharmaceutical compositions disclosed herein. Since the level of activity does not adversely affect the systemic absorption of estradiol, the formulation of the present invention gives the patient more flexibility due to the dosage regimen.

Example  13: Randomized , double Blind , Placebo control, safety results in multicenter studies.

Safety endpoints in the study included vital signs, clinical laboratory tests (blood chemistry, hematology, hormone levels, urine analysis), ECG, physical and gynecological test findings, cervical dilatation test, endometrial biopsy, and adverse events (AE) . AEs included undesirable medical conditions that occurred at any time during the entire study phase, including the period of abstinence, whether or not the study treatment was administered. AE was considered a therapeutic emergency if this occurred after study drug administration, or if it was already present and worsened during follow-up after 120 days of dosing. Participants were given a diary with instructions to record product use, sexual activity, symptoms / complaints, and other medications. AE, accompanying drugs and vital signs were recorded and evaluated at each study visit from 12 weeks of screening.

TX-004HR has a promising safety profile and is widely accepted. There was no clinically significant difference in AE between the treatment and placebo groups (Table 144). Headache was the most commonly reported TEAE followed by vaginal discharge, nasopharyngitis, and vulvar-vaginal atrophy (Table 144). Headache was the only treatment-related TEAE that was more frequent in women receiving TX-004HR than placebo (3.7% vs. 4% dose versus 3.1% for placebo). Vaginal secretions were reported by fewer women in the number of random TX-004HR groups than women in the placebo group. Most TEAEs were mild to moderate in severity. Fewer participants (1.8%) stopped study because of AE.

Nine severe TEAEs were reported in eight subjects, but none were considered to be associated with treatment. Complete heart block, appendicitis, internal organs and chronic obstructive pulmonary disease were reported by different participants in the 25 μg group, respectively. In the 10 μg group, anomalous nodular dysfunction and ankle fracture were both reported for one woman, and arthralgia and malignant melanoma were reported for one female, respectively. Women in the 4 ㎍ group did not report serious TEAE. One woman in the placebo group was reported to have cervical myelopathy. No deaths occurred during the study.

The diagnosis of endometrial hyperplasia or malignancy from endometrial biopsy was not observed at 12 weeks. Total cholesterol decreased by a mean of 0.024 mmol / l to 0.07 mmol / l in the treatment group at 12 weeks from baseline and 0.008 mmol / l in the placebo group. No clinically significant triglyceride increase was observed in any active treatment group as compared to placebo. The sex hormone binding globulin (SHBG) concentration (measured in a subset of 72 women) was not increased by treatment at 12 weeks for placebo or baseline. No clinically significant changes in laboratory parameters were found.

Phase III clinical trials demonstrate that TX-004HR at doses of 4 μg, 10 μg and 25 μg is safe and effective in treating vaginal changes and self-reported VVA symptoms in postmenopausal women. Statistically significant and clinically significant differences in all four pre-defined companion-1 endpoints (increased percentage of vaginal epithelial cells, decreased percentage of vaginal pH and vaginal epithelial cells, and decreased severity of MBS in dyspareunia) Significant improvement occurred as early as two weeks by all three doses of TX-004HR compared to placebo and lasted for 12 weeks. In addition, improvements have been found for vaginal dryness and secondary end points of vulvar or vagal irritation and itching. This improvement was achieved by systemic estrogen exposure (4 μg and 10 μg) or negligible (25 μg) without increasing systemic estrogen levels, as found in pharmacokinetic studies. TX-004HR was also highly idiomatic without any clinically significant differences observed between treatment groups and placebo groups in any AEs or treatment-related AEs and without severe AEs related to treatment.

The results demonstrate an early development of behavior in the clinical manifestations of VVA with statistically significant improved changes compared to placebo. Where the efficacy results were somewhat higher than the data from the 12 week, randomized, controlled trial comparing 10 μg of vaginal oestradiol tablets to placebo, indicating a significant reduction in the pH and percentage of epidermal and dendritic cells (See Simon et al., Obstet Gynecol. 2008; 112: 1053-1060). At week 12, the improvement was 10 times greater than that observed in this study by 10 μg of TX-004HR dose (17% in epidermal cells, -44% in basolateral cells and -1.4 in vaginal pH) (13% in epidermal cells, -37% in basolateral cells and -1.3 in vaginal pH). Although improvements in endpoints at several purposes (cell and pH) are seen by estradiol tablets within two weeks of treatment, patient reported improvements in composite scores of subjective symptoms were not observed until 8 weeks of treatment, Can be perceived as a disadvantage. This clinical trial did not evaluate individual symptoms. A second randomized controlled trial of 10 μg and 25 μg of estradiol tablets similarly found a significant improvement in the composite score of vaginal symptoms by either dose compared to placebo by 7 weeks (2 weeks, NS) of treatment I could not. Similarly, SERM, ospemifene, was evaluated in clinical trials for the treatment of dyspareunia and no statistically significant improvement was observed up to 12 weeks. Bachmann et al. Obstet Gynecol . 2008; 111: 67-76; Portman et al. Menopause. 2013; 20: 623-630.

Significantly, the results reported here showed a significant improvement in dyspareunia within two weeks by all three doses of TX-004HR by a reduction in the severity score from 1.5 to 1.7 at 12 weeks, Compared to a reduction from 1.2 to 1.6 points for reported dyspareunia. Vagifem (registered trademark) (estradiol tablets) prescription information. Nova Nordisk Pharmaceuticals Inc .; 2012; PREMARIN (registered trademark) (conjugated estrogen tablets, USP) Prescription information. Philadelphia, Pennsylvania: Wyeth Pharmaceuticals Inc .; 2010; Osphena (TM) (osmempen) tablet for oral use. Prescription information. Shionogi, Inc. See 2013.

In addition, vaginal dryness was improved from 2 weeks by 10 μg and 25 μg of TX-004HR. None of the currently available products have been reported as early onset of activity for symptoms of vaginal dryness associated with VVA, just like TX-004HR. Moreover, 10 [mu] g and 25 [mu] g of TX-004HR showed significant improvement in vaginal irritation and / or itching at 12 weeks, while none of the currently available products on the market were reported to improve this symptom. Portman et al. Maturitas . 2014; 78: 91-98; Eriksen et al. Eur J Obstet Gynecol Reprod Biol. 1992; 44: 137-144.

On the basis of large-scale postmenopausal surveys in the United States, only a small proportion (7%) of patients, possibly due to lack of information on available treatments, avoidance of discussion with health care professionals, or dissatisfaction with currently available products, ) Are considered to receive prescription valproate estrogen therapy alone for VVA (see, for example, Kingsberg et al. J Sex Med . 2013; 10: 1790-1799). Eliminating the need for an applicator or individually measuring the dose gives women a more positive user experience and thus potentially better compliance, resulting in better overall efficacy of treatment.

Results from TX-004HR in this study illustrate one of the benefits of topical vaginal estrogen therapy: rapid symptom resolution without increased systemic estrogen concentration. The mean area (AUC) and mean concentration (C _avg ) under the concentration-time curve for estradiol were not significantly different from placebo by 4 μg and 10 μg of TX-004HR. A statistically higher AUC for estradiol was observed by the 25 용량 dose, but estradiol levels were within the postmenopausal range without evidence of accumulation at 84 days. There was negligible systemic absorption, but rapid efficacy was observed within 2 weeks of dosing by all doses of TX-004HR.

The TX-004HR was very tolerant. The four most commonly reported TEAEs, including vaginal discharge and vulvar vaginal atrophy, were experienced by fewer women in the TX-004HR group than in the placebo group and mostly mild to moderate in severity. By comparison, in a 12 week study of the efficacy of osmempen, vaginal discharge was reported to be more than 6 times more common in the osmemphen group than in the placebo group (Portman et al. Menopause. 2013; 20: 623-630 ] Reference). Genital pruritus has also been reported four times more frequently in women treated with Vagifem 10 ug tablets than in placebo in a 12-month randomized study (see Vagifem TM (prescription) (Estradiol tablets) prescribing information. Novo Nordisk Pharmaceuticals Inc .; 2012). Importantly, endometrial findings after TX-004HR were benign because hyperplasia or malignancy was not reported at biopsy at 12 weeks. The occurrence of the effect was seen as fast as two weeks and persisted throughout the study. TX-004HR was very tolerant as reported here, and systemic estrogen exposure was negligible or very low as evidenced by pharmacokinetic studies.

Example  14: Randomized , double Blind , Placebo control, results of female sexual function indices in multicenter studies.

The study was a randomized, double-blind, placebo-controlled, multicenter, phase III study. Therapy was given once daily for two weeks and then twice weekly for ten weeks thereafter. (FSD) was assessed using a baseline and a multidimensional FSFI at 12 weeks. The FSFI is a simple and proven self-report questionnaire consisting of 19 questions designed to assess areas of excitement, libido, orgasm, lubrication, and pain. The index defines sexual function from a possible maximum score of 36 to a total FSFI score of 26.55 or less (sum of individual domain scores).

Postmenopausal women (aged 40 to 75 years; BMI less than 38 kg / m 2) showed up to 5% of epidermal cells in vaginal smears; A vaginal pH above 5.0; The most annoying symptom (MBS) identified by a person with moderate to severe dyspareunia; And the expected sexual activity (due to vaginal penetration) during the study period. Vulva and vaginal atrophy (VVA) treatments, including vaginal lubricants and moisturizers, were discontinued within 7 days prior to screening. The use of oral estrogen-, progestin-, androgen-, or SERM-containing drug products was banned within eight weeks of the start of the study. Changes in total and individual domain FSFI scores for each dose from baseline were compared to placebo using ANCOVA by baseline as a covariance.

764 postmenopausal women were randomized to 4 μg (n = 191), 10 μg (n = 191) or 25 μg (n = 190) vaginal estradiol soft gel capsules or placebo (n = 192) Most women were white (87%) with an average age of 59 years and an average BMI of 26.7 kg / ㎡ (Table 145). The FSFI questionnaire was completed by a person not in the PK sub-study (n = 692; 90.6%). A mean baseline value of 14.8 for all females. The overall FSFI score indicated FSD in the subjects.

The FSFI summary summary score is a descriptive summary of the numbers summarized at visits 2 and 6, and the summary summary score change (visit 6 minus visit 2) is also summarized descriptively. Changes in domain subscores and domain subscores are also summarized descriptively. The summary was based on the treatment arm and all active treatment arms were combined.

In addition, a change in the mean from the baseline of each active treatment group from the placebo group was assessed for each endpoint at a constant endpoint. A 95% CI is provided for the difference between the least squares (LS) mean change and the LS mean change between treatment and placebo. The FSFI questionnaire consists of 19 questions divided into six domains, with a minimum total score of 2.0 and a maximum score of 36.0. The FSFI questionnaire was given to the randomized population in this study, excluding this subject in the PK sub-study. At baseline, the overall mean overall score was 14.8 (14.8 for the 4 ug group, 15.8 for the 10 ug group, 14.2 for the 25 ug group, and 14.4 for the placebo group). The FSFI total score from the baseline to 12 weeks and the LS mean change in domain score are summarized in Table 146.

FSFI total scores and 12-week changes from baseline in the domains were assessed relative to placebo.

After 12 weeks, the overall FSFI score improved from baseline to a number in all groups including placebo. Overall FSFI scores were significantly increased by placebo in the 10 μg group (p <0.05) and the 25 μg group (p = 0.0019) (FIG. 24).

FSFI lubrication and pain domain scores were improved from baseline in all groups including placebo at 12 weeks, and improvements for 10 and 25 μg groups were statistically significantly higher than placebo (Fig. 25a). The 25ug composition significantly improved FSFI excitation (p = 0.0085) and satisfaction (p = 0.0073) domain scores at 12 weeks (Fig. 25b, Fig. 25c). All three doses were comparable to placebo in the effects on the FSFI domain of sexual desire and orgasm (Fig. 25d, Fig. 25e). The 4 [mu] g composition and placebo provided similar improvement levels. The composition improved the FSFI in a dose-dependent manner by a 25 [mu] g dose with the highest improvement. All three doses were effective, and the numerical improvement in subjective symptoms was the highest for subjects in the 10 and 25 μg groups. The observed placebo response can be attributed to coconut oil (Miglyol) in the formulation for placebo and estradiol compositions, which can also contribute to the observed benefits.

IX. Exemplary embodiments

Exemplary embodiments provided according to the presently disclosed subject matter include, but are not limited to, the claims and the following embodiments:

One. CLAIMS 1. A method of treating symptoms of vulvar vaginal atrophy (VVA)

Administering to a subject having VVA a vaginal suppository comprising from 4 g to 25 g of estradiol,

Wherein the treatment is effective within two weeks of the first administration.

2. The method of embodiment 1 wherein the side effect associated with administering estradiol other than headache is not significantly different from the side effects associated with administering the placebo.

3. In Embodiment 1 or Embodiment 2, the symptoms of VVA include at least one symptom selected from vaginal dryness, dyspareunia, vaginal or vulvar irritation, tingling or itching, dysuria, and vaginal bleeding associated with sexual activity.

4. The method according to any one of embodiments 1-3, comprising increasing the level of vaginal secretion in a subject as assessed by visual inspection.

5. The method according to any one of embodiments 1-4 comprising increasing the number of vaginal wrinkles in a subject as assessed by visual inspection.

6. The method according to any one of embodiments 1-5, comprising reducing vaginal bleeding or spot bleeding in a subject as assessed by visual inspection.

7. The method according to any one of embodiments 1 to 6, comprising changing the color of the vaginal mucosa in the subject from transparent to pink or from light pink to pink, as assessed by visual inspection.

8. The method of any one of embodiments 1-7, wherein the treatment reduces the severity of vaginal dryness within two weeks.

9. The method according to any one of embodiments 1 to 8, wherein the treatment reduces the severity of vulvar or vaginal itching within two weeks.

10. The method according to any one of embodiments 1-9, wherein the treatment reduces the severity of dyspareunia within two weeks.

11. The composition of any one of embodiments 1 to 10, wherein the vaginal supplements further comprise a solubilizer, wherein the solubilizer comprises at least one C6-C12 fatty acid or glycol, Lt; / RTI &gt; ester.

12. The method of any one of embodiments 1-11, wherein the vaginal suppository comprises 4 ug of estradiol.

13. In Embodiment 12, administration of a vaginal suppository comprises, in a plasma sample from a patient,

1) an unadjusted arithmetic mean area (AUC) of the estradiol in the range of about 73.3 pg * hr / ml to about 114.7 pg * hr / ml when assessed on day 1 _0-24 ;

2) a corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 3.1 pg / ml to about 4.8 pg / ml when assessed on day 1;

3) an unadjusted arithmetic mean area (AUC) of the estradiol in the range of about 69.7 pg * hr / ml to about 108.9 pg * hr / ml when assessed on day 14 _0-24 ; And

4) provide at least one parameter selected from the corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 2.8 pg / ml to about 4.6 pg / How to.

13. The method of embodiment 12 wherein administering a vaginal suppository comprises administering to the patient a plasma corrected sample geometric mean peak of estradiol in the range of from about 2.0 pg / ml to about 3.3 pg / 0.0 &gt; ( _Cmax ). &_Lt; / RTI &gt;

14. The method of embodiment 12 or 13 wherein administering a vaginal suppository comprises administering to the patient a plasma sample in the range of about 1.0 pg * hr / ml to about 1.7 pg * hr / To provide a corrected geometric mean peak plasma concentration (C _max ) of estradiol.

15. The method of any one of embodiments 12-14, wherein administering the vaginal suppository comprises administering to the patient a plasma sample in a range of about 9.5 pg * hr / ml to about 15.1 pg * hr / To provide a corrected curve geometric mean area (AUC) of _0-24 of estradiol.

16. The method of any one of embodiments 12-15, wherein administering a vaginal suppository comprises administering to the patient a plasma sample in a range of about 5.7 pg * hr / ml to about 9.1 pg * hr / To provide a corrected curve geometric mean area (AUC) of _0-24 of estradiol.

17. 11. The method according to any one of embodiments 1-11, wherein the vaginal suppository comprises 10 g of estradiol.

19. In Embodiment 17, administration of a vaginal suppository can be performed in a plasma sample from a patient,

1) an unadjusted arithmetic mean area (AUC) of _0-24 of estradiol in the range of about 110.3 pg * hr / ml to about 172.6 pg * hr / ml when assessed on day 1;

2) a corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 4.6 pg / ml to about 7.8 pg / ml when assessed on day 1;

3) Unadjusted arithmetic mean area (AUC) _0-24 of estradiol in the range of about 87.9 pg * hr / ml to about 137.4 pg * hr / ml when assessed on day 14; And

4) provide at least one parameter selected from the corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 3.6 pg / ml to about 5.8 pg / How to.

18. The method of embodiment 17, wherein administering a vaginal suppository comprises administering to the patient a plasma of a calorimetric average peak of estradiol in the range of about 4.7 pg / ml to about 7.6 pg / 0.0 &gt; ( _Cmax ). &_Lt; / RTI &gt;

19. The method of embodiment 17 or 18 wherein administering a vaginal suppository comprises administering to a plasma sample from a patient a calibration of estradiol in the range of about 2.3 pg / ml to about 3.8 pg / ml when evaluated at 14 days (C _max ), &lt; / RTI &gt;

20. The method of any one of embodiments 17-19, wherein administering the vaginal suppository comprises administering to the patient a plasma sample in the range of about 17.5 pg * hr / ml to about 27.4 pg * hr / To provide a corrected curve geometric mean area (AUC) of _0-24 of estradiol.

21. The method of any one of embodiments 17-20, wherein administering the vaginal suppository comprises administering to the patient a plasma sample in the range of about 10.9 pg * hr / ml to about 17.2 pg * hr / To provide a corrected curve geometric mean area (AUC) of _0-24 of estradiol.

22. The method of any one of embodiments 17-21, wherein administering vaginal suppositories comprises administering estradiol in a plasma sample from a patient in the range of about 0.6 pg / ml to about 1.1 pg / And providing a corrected geometric mean peak plasma concentration (C _avg ).

23. The method of any one of embodiments 17-22, wherein administering vaginal suppositories comprises administering an amount of estradiol in a plasma sample from a patient in the range of about 0.1 pg / ml to about 0.3 pg / And providing a corrected geometric mean peak plasma concentration (C _avg ).

24. The method of any one of embodiments 1-11, wherein the vaginal suppository comprises 25 ug of estradiol.

27. In Embodiment 24, administration of a vaginal suppository may be such that, in a plasma sample from a patient,

1) an unadjusted arithmetic mean area (AUC) _0-24 of estradiol in the range of about 173.5 pg * hr / ml to about 271.3 pg * hr / ml when evaluated on day 1;

2) a corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 7.2 pg / ml to about 11.4 pg / ml when assessed on day 1;

3) an unadjusted arithmetic mean area (AUC) of the estradiol in the range of about 137.5 pg * hr / ml to about 215.1 pg * hr / ml when assessed on day 14 _0-24 ; And

4) provide at least one parameter selected from the corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 5.7 pg / ml to about 9.0 pg / How to.

25. The method of embodiment 24 wherein administering a vaginal suppository comprises administering to the patient a plasma of a calibrated geometric mean peak of estradiol in the range of about 20.9 pg / ml to about 32.8 pg / 0.0 &gt; ( _Cmax ). &_Lt; / RTI &gt;

26. The method of embodiment 24 or embodiment 25 wherein administering a vaginal suppository comprises administering to the patient a plasma sample that has a calibrator of estradiol in the range of about 9.5 pg / ml to about 15.1 pg / (C _max ), &lt; / RTI &gt;

27. The method as in any one of embodiments 24-26, wherein administering the vaginal suppository comprises administering to the patient a plasma sample in a range of about 104.3 pg * hr / ml to about 163.1 pg * hr / To provide a corrected curve geometric mean area (AUC) of _0-24 of estradiol.

28. The method of any one of embodiments 24-27, wherein administering a vaginal suppository comprises administering to the patient a plasma sample in the range of about 67.6 pg * hr / ml to about 105.8 pg * hr / To provide a corrected curve geometric mean area (AUC) of _0-24 of estradiol.

29. The method of any one of embodiments 24-28, wherein administering a vaginal suppository comprises administering an amount of estradiol in a plasma sample from a patient in the range of about 4.3 pg / ml to about 6.8 pg / And providing a corrected geometric mean peak plasma concentration (C _avg ).

30. The method of any one of embodiments 24-29, wherein administering a vaginal suppository comprises administering an amount of estradiol in a plasma sample from a patient in the range of about 2.7 pg / ml to about 4.4 pg / And providing a corrected geometric mean peak plasma concentration (C _avg ).

31. The method as in any one of embodiments 12-30, wherein administering a vaginal suppository comprises adjusting the corrected geometric mean time to peak plasma concentration of estradiol in the plasma sample from the patient of from about 0.25 hours to about 2 hours, _max. &lt; / RTI &gt;

32. The method according to any one of embodiments 1 to 31, wherein the vaginal suppository does not comprise a hydrophilic gel-forming bioadhesive in the solubilizer.

33. 31. The method of any one of embodiments 1-32, wherein the estradiol is the only active hormone in the vaginal suppository.

34. 33. The method according to any one of embodiments 1-33, wherein administration is performed twice daily for two weeks and thereafter every week.

35. 33. The method according to any one of embodiments 1-33, wherein the subject is on gait for a period of time beginning at about 5 minutes after administration of the vaginal suppository and ending at about 4 hours after administration of the vaginal suppository.

36. CLAIMS What is claimed is: 1. A method of treating female sexual dysfunction, comprising administering to a female subject in need thereof a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 microgram of estradiol; Estradiol is the only active hormone in the vaginal suppository.

37. The method of embodiment 36, wherein the vaginal suppository does not comprise a hydrophilic gel-forming bioadhesive in the liquid composition.

38. 36. The method of embodiment 36, wherein treating female sexual function comprises treating an animal's sexual desire, excitement, lubrication, satisfaction, and / or orgasm.

39. The method of embodiment 38, wherein the treatment is assessed using a female sexual function index.

40. Embodiment 36. The method of embodiment 36, wherein the suppository is administered to the subject twice daily for two weeks and thereafter twice a week.

41. The method of embodiment 36, wherein the suppository comprises about 4 g of estradiol.

42. The method of embodiment 36, wherein the suppository comprises about 10 [mu] g of estradiol.

43. The method of embodiment 36, wherein the suppository comprises about 25 μg of estradiol.

44. CLAIMS What is claimed is: 1. A method of treating vaginal dryness comprising administering to a female subject in need thereof a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 microgram of estradiol; Estradiol is the only active hormone in the vaginal suppository.

45. The method of embodiment 44, wherein the vaginal suppository does not include a hydrophilic gel-forming bioadhesive in the liquid composition.

46. Embodiment 44. The method of embodiment 44, wherein the suppository is administered to the subject twice daily for two weeks and thereafter every two weeks.

47. The method of embodiment 44, wherein the suppository comprises about 4 g of estradiol.

48. The method of embodiment 44, wherein the suppository comprises about 10 μg of estradiol.

49. The method of embodiment 44, wherein the suppository comprises about 25 μg of estradiol.

50. The method of embodiment 44, wherein the treatment reduces vaginal dryness within two weeks.

51. A method of treating dyspareunia comprising administering to a female subject in need of treatment of dyspareunia (a) a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 microgram of estradiol; Estradiol is the only active hormone in the vaginal suppository.

52. The method of embodiment 51, wherein the vaginal suppository does not comprise a hydrophilic gel-forming bioadhesive in the liquid composition.

53. Embodiment 51. The method of embodiment 51, wherein the suppository is administered to the subject twice daily for two weeks and thereafter every two weeks.

54. Embodiment 51. The method of embodiment 51, wherein the suppository comprises about 4 g of estradiol.

55. Embodiment 51. The method of embodiment 51, wherein the suppository comprises about 10 [mu] g of estradiol.

56. The method of embodiment 51, wherein the suppository comprises about 25 μg of estradiol.

57. Embodiment 51. The method of embodiment 51, wherein the treatment reduces dyspareunia within two weeks.

58. A method of reestrogenating a vagina, labia or vulva, comprising: (a) administering to a female subject a vaginal, labial or vulvar reestrogenation requiring a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 microgram of estradiol; Estradiol is the only active hormone in the vaginal suppository.

59. Embodiment 58. The method of embodiment 58, wherein the suppository is administered to the subject twice daily for two weeks and thereafter twice a week.

60. The method of embodiment 58, wherein the suppository comprises about 4 g of estradiol.

61. The method of embodiment 58, wherein the suppository comprises about 10 [mu] g of estradiol.

62. The method of embodiment 58, wherein the suppository comprises about 25 μg of estradiol.

63. The method of embodiment 58, wherein the vaginal suppository does not comprise a hydrophilic gel-forming bioadhesive in the liquid composition.

While pharmaceutical compositions and methods are described in terms of what is presently believed to be a practical and preferred embodiment, it should be understood that this disclosure is not necessarily limited to the disclosed embodiments. This is intended to cover various modifications and similar arrangements included within the spirit and scope of the claims, and the scope of the claims should be accorded the broadest interpretation so as to encompass all such modifications and similar embodiments. The present disclosure encompasses any and all embodiments of the following claims.

Claims (66)

A method of treating the symptoms of vulvovaginal atrophy (VVA)
Administering to a subject having VVA a vaginal suppository comprising from 4 g to 25 g of estradiol,
Wherein said treatment is effective within two weeks of said first administration. The method of claim 1, wherein the side effect associated with administering estradiol other than headache is not significantly different from the side effects associated with administering the placebo. 3. The method according to claim 1 or 2, wherein the symptoms of VVA include at least one symptom selected from vaginal dryness, dyspareunia, vaginal or vulvar irritation, tingling or itching, dysuria and vaginal bleeding associated with sexual activity , Vocal-atrophy (VVA). The method of claim 1, comprising increasing the level of vaginal secretion in the subject, as assessed by visual inspection, treating the symptoms of the vulvar-vaginal atrophy (VVA). The method of claim 1, comprising increasing the number of vaginal wrinkles in the subject as assessed by visual inspection. 3. The method of claim 1, comprising reducing vaginal bleeding or petechiae in the subject as assessed by visual inspection. The method according to claim 1, further comprising the step of changing the color of the vaginal mucosa in the subject from transparent to pink or from light pink to pink, as assessed by visual inspection, How to cure. 2. The method of claim 1, wherein the treatment reduces the severity of vaginal dryness within two weeks. 2. The method of claim 1 wherein the treatment reduces the severity of vulvar or vaginal itching within two weeks. 2. The method of claim 1, wherein the treatment reduces the severity of dyspareunia within two weeks. 3. The composition of claim 1, wherein the vaginal suppository further comprises a solubilizer, wherein the solubilizer comprises at least one C6-C12 fatty acid or glycol, monoglyceride, diglyceride or triglyceride ester thereof Gt; (VVA) &lt; / RTI &gt; 2. The method of claim 1 wherein the vaginal suppository comprises 4 g of estradiol. 13. The method of claim 12, wherein administering the vaginal suppository comprises:
1) an unadjusted arithmetic mean area (AUC) of the estradiol in the range of about 73.3 pg * hr / ml to about 114.7 pg * hr / ml when assessed on day 1 _0-24 ;
2) a corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 3.1 pg / ml to about 4.8 pg / ml when assessed on day 1;
3) an unadjusted arithmetic mean area (AUC) of the estradiol in the range of about 69.7 pg * hr / ml to about 108.9 pg * hr / ml when assessed on day 14 _0-24 ; And
4) provide at least one parameter selected from the corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 2.8 pg / ml to about 4.6 pg / Gt; (VVA) &lt; / RTI &gt; 13. The method of claim 12, wherein administration of the vaginal suppository comprises administering to the patient a plasma of a calibrated geometric mean peak of estradiol in the range of about 2.0 pg / ml to about 3.3 pg / A method of treating a symptom of vulvar vaginal atrophy (VVA), which provides a plasma concentration (C _max ). 13. The method of claim 12, wherein administration of the vaginal suppository comprises administering to the plasma sample from the patient a calibration of estradiol in the range of about 1.0 pg * hr / ml to about 1.7 pg * hr / Gt; (Vma) &lt; / RTI &gt; that provides a geometric mean peak plasma concentration ( _Cmax ). 13. The method of claim 12, wherein administering the vaginal suppository comprises administering to the plasma sample from the patient a calibration of estradiol in the range of about 9.5 pg * hr / ml to about 15.1 pg * hr / (VVA), which provides a geometric mean area under the curve (AUC) _0-24 . 13. The method of claim 12, wherein administration of the vaginal suppository comprises: in a plasma sample from the subject, a correction of estradiol in the range of about 5.7 pg * hr / ml to about 9.1 pg * hr / (VVA), which provides a geometric mean area under the curve (AUC) _0-24 . 2. The method of claim 1 wherein the vaginal suppository comprises 10 g of estradiol. 19. The method of claim 18, wherein administering the vaginal suppository comprises, in a plasma sample from the patient,
1) an unadjusted arithmetic mean area (AUC) of _0-24 of estradiol in the range of about 110.3 pg * hr / ml to about 172.6 pg * hr / ml when assessed on day 1;
2) a corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 4.6 pg / ml to about 7.8 pg / ml when assessed on day 1;
3) Unadjusted arithmetic mean area (AUC) _0-24 of estradiol in the range of about 87.9 pg * hr / ml to about 137.4 pg * hr / ml when assessed on day 14; And
4) provide at least one parameter selected from the corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 3.6 pg / ml to about 5.8 pg / Gt; (VVA) &lt; / RTI &gt; 19. The method of claim 18, wherein administering the vaginal suppository comprises administering to the patient a plasma of a calibrated geometric mean peak of estradiol in the range of about 4.7 pg / ml to about 7.6 pg / A method of treating a symptom of vulvar vaginal atrophy (VVA), which provides a plasma concentration (C _max ). 19. The method of claim 18, wherein the administering of the vaginal suppository comprises, in a plasma sample from the subject, a calibrated geometric mean peak of estradiol in the range of about 2.3 pg / ml to about 3.8 pg / A method of treating a symptom of vulvar vaginal atrophy (VVA), which provides a plasma concentration (C _max ). 19. The method of claim 18, wherein administering the vaginal suppository comprises administering to the plasma sample from the patient a calibration of estradiol in the range of about 17.5 pg * hr / ml to about 27.4 pg * hr / (VVA), which provides a geometric mean area under the curve (AUC) _0-24 . 19. The method of claim 18, wherein administering the vaginal suppository comprises administering to the plasma sample from the patient a calibration of estradiol in the range of about 10.9 pg * hr / ml to about 17.2 pg * hr / (VVA), which provides a geometric mean area under the curve (AUC) _0-24 . 19. The method of claim 18, wherein administering the vaginal suppository comprises administering to the patient a plasma of a calibrated geometric mean peak of estradiol in the range of about 0.6 pg / ml to about 1.1 pg / A method of treating symptoms of a vulvar vaginal atrophy (VVA) that provides plasma concentration (C _avg ). 19. The method of claim 18, wherein the administering of the vaginal suppository comprises, in a plasma sample from the patient, a calibrated geometric mean peak of estradiol in the range of about 0.1 pg / ml to about 0.3 pg / A method of treating symptoms of a vulvar vaginal atrophy (VVA) that provides plasma concentration (C _avg ). 2. The method of claim 1 wherein the vaginal suppository comprises 25 g of estradiol. 27. The method of claim 26, wherein administering the vaginal suppository comprises, in a plasma sample from the patient,
1) an unadjusted arithmetic mean area (AUC) _0-24 of estradiol in the range of about 173.5 pg * hr / ml to about 271.3 pg * hr / ml when evaluated on day 1;
2) a corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 7.2 pg / ml to about 11.4 pg / ml when assessed on day 1;
3) an unadjusted arithmetic mean area (AUC) of the estradiol in the range of about 137.5 pg * hr / ml to about 215.1 pg * hr / ml when assessed on day 14 _0-24 ; And
4) provide at least one parameter selected from the corrected arithmetic mean peak plasma concentration (C _{avg [0-24]} ) of estradiol in the range of about 5.7 pg / ml to about 9.0 pg / Gt; (VVA) &lt; / RTI &gt; 27. The method of claim 26, wherein the administering of the vaginal suppository comprises administering to the patient a plasma of a calibrated geometric mean peak of estradiol in the range of about 20.9 pg / ml to about 32.8 pg / A method of treating a symptom of vulvar vaginal atrophy (VVA), which provides a plasma concentration (C _max ). 27. The method of claim 26, wherein the administering of the vaginal suppository comprises administering, in a plasma sample from the patient, a calibrated geometric mean peak of estradiol in the range of about 9.5 pg / ml to about 15.1 pg / A method of treating a symptom of vulvar vaginal atrophy (VVA), which provides a plasma concentration (C _max ). 27. The method of claim 26, wherein administration of the vaginal suppository comprises administering to the plasma sample from the patient an amount of estradiol in the range of from about 104.3 pg * hr / ml to about 163.1 pg * hr / (VVA), which provides a geometric mean area under the curve (AUC) _0-24 . 27. The method of claim 26, wherein administering the vaginal suppository comprises administering to the plasma sample from the patient a calibration of estradiol in the range of about 67.6 pg * hr / ml to about 105.8 pg * hr / (VVA), which provides a geometric mean area under the curve (AUC) _0-24 . 27. The method of claim 26, wherein administering the vaginal suppository comprises administering to the patient a plasma of a calibrated geometric mean peak of estradiol in the range of about 4.3 pg / ml to about 6.8 pg / A method of treating symptoms of a vulvar vaginal atrophy (VVA) that provides plasma concentration (C _avg ). 27. The method of claim 26, wherein administering the vaginal suppository comprises administering to the patient a plasma of a calorimetric average peak of estradiol in the range of about 2.7 pg / ml to about 4.4 pg / A method of treating symptoms of a vulvar vaginal atrophy (VVA) that provides plasma concentration (C _avg ). 27. The method of claim 26, wherein administration of the vaginal suppository comprises providing a corrected geometric mean time ( _Tmax ) for a peak plasma concentration of estradiol in a plasma sample from the patient of about 0.25 hours to about 2 hours , Vocal-atrophy (VVA). The method of claim 1, wherein the vaginal suppository does not include a bioadhesive agent in the solubilizing agent. 8. The method of claim 1, wherein the estradiol is the only active hormone in the vaginal suppository, wherein the symptom of the vulvar-vaginal atrophy (VVA) is treated. 2. The method of claim 1, wherein said administration is performed twice daily for two weeks and thereafter every two weeks for the treatment of symptoms of vulvar-vaginal atrophy (VVA). 4. The method of claim 1, wherein the subject is at least one of the following: vocal-vaginal atrophy (VVA), which is ongoing during a period of time beginning at about 5 minutes after administration of the vaginal suppository and ending at about 4 hours after administration of the vaginal suppository How to treat symptoms. CLAIMS What is claimed is: 1. A method of treating female sexual dysfunction, comprising administering to a female subject in need thereof a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 micrograms of estradiol; Wherein estradiol is the only active hormone in the vaginal suppository. 40. The method of claim 39, wherein the vaginal suppository does not include a hydrophilic gel-forming bioadhesive in the liquid composition. 40. The method of claim 39, wherein treating the female sexual function comprises treating the sexual desire, excitement, lubrication, satisfaction, and / or orgasm of the subject. 42. The method of claim 41, wherein the treatment is assessed using a Female Sexual Function Index. 40. The method of claim 39, wherein said suppository is administered to said subject twice daily for two weeks and thereafter twice a week. 40. The method of claim 39, wherein the suppository comprises about 4 g of estradiol. 40. The method of claim 39, wherein said suppository comprises about 10 [mu] g of estradiol. 40. The method of claim 39, wherein the suppository comprises about 25 g of estradiol. CLAIMS What is claimed is: 1. A method of treating vaginal dryness comprising administering to a female subject in need thereof a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 micrograms of estradiol; Wherein estradiol is the only active hormone in the vaginal suppository. 50. The method of claim 47, wherein the vaginal suppository does not comprise a hydrophilic gel-forming bioadhesive in the liquid composition. 48. The method of claim 47, wherein said suppository is administered to said subject twice daily for two weeks and thereafter twice a week. 48. The method of claim 47, wherein said suppository comprises about 4 g of estradiol. 50. The method of claim 47, wherein the suppository comprises about 10 [mu] g of estradiol. 50. The method of claim 47, wherein said suppository comprises about 25 &lt; RTI ID = 0.0 &gt; g &lt; / RTI &gt; of estradiol. 48. The method of claim 47, wherein said treatment reduces vaginal dryness within two weeks. A method of treating dyspareunia comprising administering to a female subject in need of treatment of dyspareunia (a) a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 micrograms of estradiol; Wherein estradiol is the only active hormone in the vaginal suppository. 55. The method of claim 54, wherein the vaginal suppository does not include a hydrophilic gel-forming bioadhesive in the liquid composition. 55. The method of claim 54, wherein said suppository is administered to said subject twice daily for two weeks and thereafter twice a week. 55. The method of claim 54, wherein said suppository comprises about 4 g of estradiol. 55. The method of claim 54, wherein the suppository comprises about 10 [mu] g of estradiol. 55. The method of claim 54, wherein said suppository comprises about 25 g of estradiol. 55. The method of claim 54, wherein said treatment reduces dyspareunia within two weeks. A method of reestrogenating a vagina, labia or vulva, comprising: (a) administering to a female subject a vaginal, labial or vulvar reestrogenation requiring a therapeutically effective amount of estradiol; Caprylic / capry triglyceride; A liquid composition comprising a non-ionic surfactant comprising PEG-6 palmitostearate and ethylene glycol palmitostearate; And (b) a soft suppository comprising soft gelatin capsules, wherein the vaginal suppository comprises from about 1 microgram to about 25 micrograms of estradiol; Estradiol is the only active hormone in the vaginal suppository that reestrogenates vagina, labia or vulva. 62. The method of claim 61, wherein said suppository is administered to said subject twice daily for two weeks and thereafter twice per week. 63. The method of claim 61, wherein the suppository comprises about 4 g of estradiol. 62. The method of claim 61, wherein the suppository comprises about 10 [mu] g of estradiol. 62. The method of claim 61, wherein the suppository comprises about 25 ug of estradiol. 62. The method of claim 61, wherein the vaginal suppository does not comprise a hydrophilic gel-forming bioadhesive in the liquid composition.

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