US 2011 0098.258A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0098258 A1 Masini-Eteve et al. (43) Pub. Date: Apr. 28, 2011

(54) TRANSDERMAL PHARMACEUTICAL (30) Foreign Application Priority Data COMPOSITIONS COMPRISING ACTIVE AGENTS Dec. 10, 2009 (EP) ...... 09 178762.2 Publication Classification (75) Inventors: Yalers Mility, is Canet (51) Int. Cl. site ); Denis Canet, A6II 3/56 (2006.01) russels (BE) A6II 3/566 (2006.01) A 6LX 3/573 (2006.01) (73) Assignee: Besins Healthcare Luxembourg A6II 3/5685 (2006.01) (52) U.S. Cl...... 514/170; 514/169; 514/182: 514/177; (21) Appl. No.: 12/912,310 514/178 (57) ABSTRACT (22) Filed: Oct. 26, 2010 The present invention provides compositions and methods for Related U.S. Application Data providing sustained release of an active agent through the skin of a Subject, wherein a pharmaceutical percutaneous (60) Provisional application No. 61/255.241, filed on Oct. composition comprises at least one fatty acid ester and a 27, 2009. therapeutically effective amount of active agent.

1.00 Ethyl Oleate, % 50 .5 39

Estradiol, %

Effect of the Ethyl Oleate and Estradiol concentration on the total penetration of Estradiol over 48 hours. Patent Application Publication Apr. 28, 2011 Sheet 1 of 10 US 2011/00982.58A1

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Figure 1. The amount of progesterone delivered through the skin in 48 hours for the different formulations tested in Example 5. Patent Application Publication Apr. 28, 2011 Sheet 2 of 10 US 2011/00982.58A1

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Figure 2. The amount of estradiol delivered through the skin in 48 hours relative to the drug loading in the formulation of the invention (diamonds) in comparison with Estroge (0.06%) shown as a square. Patent Application Publication Apr. 28, 2011 Sheet 3 of 10 US 2011/00982.58A1

.5

15 39 0.04 Estradiol, %

Figure 3. Effect of the Oleic Acid, Propylene Glyco and Estradio concentration on the total penetration of Estradiol over 48 hours. Patent Application Publication Apr. 28, 2011 Sheet 4 of 10 US 2011/00982.58A1

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Figure 9. Time/Flux profile for 5 experimental data points. Patent Application Publication Apr. 28, 2011 Sheet 10 of 10 US 2011/0098258A1

2, 5

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EtOH (96% viv) concentration in the mixture

Figure 10. Ethyl Oleate solubility as a function of Ethanol concentration in a mixture containing 0.24% Estradiol, 5% propylene glycol and 2% oleic acid. US 2011/0098258 A1 Apr. 28, 2011

TRANSIDERMAL PHARMACEUTICAL ally occurs many hours later, or once-a-day. Such a sudden COMPOSITIONS COMPRISING ACTIVE rise of drug concentration in the blood could be dangerous for AGENTS the patient as it may exceed the drug dose tolerated by the organism. In addition, as the whole dose of active agent is PRIORITY delivered to the bloodstream and tissues in the first hours 0001. This application claims the benefit under 35 U.S.C. following the application, the drug's intended effects may not S119(e) of U.S. Provisional Application No. 61/255.241 filed endure until the next application. on Oct. 27, 2009, the entire contents of which are incorpo 0006. There remains a need, therefore, for transdermal rated herein by reference in their entirety, and also claims the pharmaceutical compositions capable of delivering at least a benefit under 35 U.S.C. S119(a) of European Application No. part of their active content in a controlled-release manner, for EP 09 178762.2, filed on Dec. 10, 2009. example through temporary storage in the dermis. FIELD OF THE INVENTION SUMMARY 0002 Disclosed herein are compositions and methods for 0007. Described herein are sustained release transdermal delivering a therapeutically active agent through the skin of a pharmaceutical compositions, and methods of making and Subject, e.g., to transdermal pharmaceutical compositions. using them. 0008. In accordance with some embodiments, there are BACKGROUND provided Sustained release pharmaceutical compositions for 0003. It is well known that certain therapeutically active topical administration to a skin Surface comprising: a phar agents are not suitable for oral administration for various maceutically active agent comprising one or more steroids, a reasons associated, inter alia, with either a high level of fatty acid ester, water, a C2-C6 monoalcohol and a fatty acid, metabolism in the liver (“first pass effect”) or a high level of wherein the weight: weight ratio of the fatty acid ester in the gastrointestinal degradation. Transdermal or transmucosal composition to the total active agent in the composition is at formulations have been developed in order to circumvent least 4:1 fatty acid ester:active agent. In some embodiments, these drawbacks. Specifically, pharmaceutical compositions the weight:weight ratio offatty acid ester:active agent in the for transdermal or transmucosal administration have several composition ranges from about 4:1 to about 20:1. advantages over oral forms, including elimination of the 0009. In some embodiments, the composition further problems associated with metabolism of the therapeutically comprises a co-solvent, Such as propylene glycol. In some active agent by the liver and with gastric degradation of the embodiments, the co-solvent is present in an amount ranging active agent. However, transdermal and transmucosal com from 0.01% to 7%, by weight of the total weight of the positions face problems associated with the kinetics of pas pharmaceutical composition, or in an amount ranging from sage of the therapeutically active agents from the Surface of 3% to 7% by weight of the total weight of the pharmaceutical the skin into the bloodstream. composition. 0004 Indeed, the skin is a heterogeneous tissue which 0010. In some embodiments, the fatty acid ester is selected comprises two layers: the dermis and the outer most epider from the group consisting of ethyl oleate, isopropyl oleate, mis layer, which can be further divided into the stratum cor isopropyl myristate, isopropyl isostearate, isopropyl palmi neum and the viable epidermis. These layers provide the skin tate, ethyl octanoate, ethyl dodecanoate, ethyllinoleate, ethyl with barrier capacities against the entry of foreign Substances palmitoleate, ethyl isostearate and ethyl linolenate. In some Such as drugs. The stratum corneum acts as a physical diffu embodiments, the fatty acid ester is the ester that would result sive barrier, whereas the epidermis and dermis can provide in from the reaction of the fatty acid formulated in the compo addition a biochemical or enzymatic barrier. sition with an alcohol. In other embodiments, the fatty acid 0005 Studies concerning the absorption of therapeuti ester is not the ester that would result from the reaction of the cally active agents by the skin have focused for the most part fatty acid formulated in the composition with the alcohol on improving the rate of absorption of the active agents ingre formulated in the composition. In some embodiments, the dients through the skin, rather than paying any attention to the fatty acid ester is present in an amount ranging from 0.01% to fate of the absorbed active agents. For example, the use of 5% by weight of the total weight of the pharmaceutical com permeation enhancers was proposed in order to increase the position, or in an amount ranging from 0.05% to 2.4% by initial rate of penetrated active agents through the skin. The weight of the total weight of the pharmaceutical composition, term "permeation enhancer generally refers to any molecule or in an amount ranging from 0.1% to 2.2% by weight of the that promotes the reversible diffusion of an active agent total weight of the pharmaceutical composition. through the skin or mucous membranes, and any solubilizing 0011. In some embodiments, the fatty acid is a C8-C22 agent that promotes the partitioning of the active agent fatty acid. In some embodiments, the fatty acid is selected between the vehicle and the horny layer of the epidermis or of from the group consisting of capric acid, lauric acid, myristic the mucous membranes. Most enhancers affect the stratum acid, palmitic acid, Stearic acid, oleic acid, isostearic acid, corneum barrier capacities, i.e., they reversibly alter the Stra palmitoleic acid, linoleic acid and linolenic acid. For tum corneum structure, thus increasing drug diffusivity and example, in some embodiments, the fatty acid is oleic acid. In solubility. This indeed enhances skin penetration of the active Some embodiments, the fatty acid is present in an amount agents, but this may also result in the direct absorption of a ranging from 0.01% to 5% by weight of the total weight of the large amount of the drug through the tissues, leading to a peak pharmaceutical composition, or in an amount ranging from active agent concentration in the blood in the immediate 0.05% to 3.5% by weight of the total weight of the pharma hours following the application of the composition. This ini ceutical composition, or in an amount ranging from 1.0% to tial peak is often followed by a trough in blood concentrations 3.0% by weight of the total weight of the pharmaceutical prior to the next application of the composition, which usu composition. US 2011/0098258 A1 Apr. 28, 2011

0012. In specific embodiments, compositions comprise In yet other embodiments Sustained release of the pharma 2% ethyl oleate as the fatty acid ester, 2% oleic acid as the ceutically active agent through the skin is observed at least 48 fatty acid, and 5% propylene glycol as the co-solvent, all by hours after its administration. weight of the total weight of the pharmaceutical composition. In other specific embodiments, compositions comprise 0.3% BRIEF DESCRIPTION OF THE DRAWINGS ethyl oleate as the fatty acid ester, 0.3% oleic acid as the fatty 0018 FIG. 1 represents the amount of progesterone deliv acid, and 0.75% propylene glycol as the co-solvent, all by ered through the skin in 48 hours (flux, ug/cm2/hr) for the weight of the total weight of the pharmaceutical composition. different formulations tested in Example 5. ( Formulation 0013. In some embodiments, the pharmaceutically active 1; O—Formulation 2: A Progestogel (1% progesterone agent is selected from the group consisting of estrogens, hydroalcholic gel) (Besins Healthcare)). anti-estrogens (or SERMs), androgens, anti-androgens, 0019 FIG. 2 represents the amount of estradiol delivered progestins, and mixtures thereof. In some embodiments, the (ug) through the skin in 48 hours relative to the drug loading pharmaceutically active agent is selected from estradiol and in one exemplary formulation (diamonds) and in comparison progesterone, and the fatty acid ester is ethyl oleate. In other to Estrogel(R) (0.06% estradiol gel, square) (Ascend Thera specific embodiments, the pharmaceutically active agent is peutics). selected from testosterone and dihydrotestosterone (DHT), 0020 FIG.3 represents the effect of the oleic acid, propy lene glycol and estradiol concentrations on the total penetra and the fatty acid ester is selected from ethyl oleate and tion of estradiol (ug) over 48 hours. isopropyl myristate. In some embodiments, the active agentis 0021 FIG. 4 represents the effect of the oleic acid, propy present in an amount ranging from 0.01% to 5% by weight of lene glycol and estradiol concentrations on the total penetra the total weight of the pharmaceutical composition. tion of estradiol (ug) over 48 hours. 0014. In some embodiments, the alcohol is selected from (0022 FIG. 5 represents the effect of the ethyl oleate and the group consisting of ethanol, n-propanol, isopropanol, estradiol concentrations on the total penetration of estradiol n-butanol, isobutanol, tert-butanol, and mixtures thereof. For (ug) over 48 hours. example, in Some embodiments, the alcohol is ethanol. In (0023 FIG. 6 represents the effect of the ethyl oleate and Some embodiments, the alcohol is present in an amount rang estradiol concentrations on the total penetration of estradiol ing from 10% to 90% by weight of the total weight of the (ug) over 48 hours. pharmaceutical composition, or in an amount ranging from 0024 FIG. 7 represents the flux profile (ug/cm2/hr) over 20% to 80% by weight of the total weight of the pharmaceu time for three compositions tested in Example 7. ( For tical composition, or in an amount ranging from 45% to 75% mulation 301: O Formulation 303: A Formulation 309.) by weight of the total weight of the pharmaceutical compo 0025 FIG. 8 represents the flux profile (ug/cm2/hr) over sition. time for three compositions tested in Example 7. ( For 0.015. In accordance with other embodiments there are mulation 306; O. Formulation 307: A Formulation 311.) provided methods of making a Sustained release pharmaceu 0026 FIG. 9 represents the flux profile (ug/cm2/hr) over tical composition for topical administration to a skin Surface time for five compositions tested in Example 7. ( Formu comprising mixing a pharmaceutically active agent compris lation 302; O. Formulation 304: A Formulation 305; ing one or more steroids, a fatty acid ester, water, a C2-C6 V Formulation 308; —Formulation 310.) monoalcohol and a fatty acid, wherein the weightweight (0027 FIG. 10 represents the solubility of ethyl oleate ratio of the fatty acid ester in the composition to the total (g/100g) as a function of ethanol (96%) concentration (v/v) in active agent in said composition is at least 4:1 fatty acid a mixture containing 0.24% estradiol, 5% propylene glycol ester:active agent. In some embodiments, the weight: weight and 2% oleic acid, all by weight of the total weight of the ratio offatty acid ester:active agent in the composition ranges composition. from about 4:1 to about 20:1. Any composition as described above and below may be made by such methods. DETAILED DESCRIPTION 0016. In accordance with other embodiments there are 0028 Few studies have investigated whether transder provided methods for providing a Sustained release of a phar mally administered active agents pass directly through the maceutically active agent through the skin of a Subject, com skin into the bloodstream, or whether they are first retained prising topically administering to the skin of the Subject a within a compartment within the skin that serves as an active pharmaceutical composition comprising a pharmaceutically agent storage depot, prior to being released into the circula active agent comprising one or more steroids, a fatty acid tion. It is known from the article entitled “Will cutaneous ester, water, a C2-C6 monoalcohol and a fatty acid, wherein levels of absorbed material be systemically absorbed? the weight: weight ratio of the fatty acid ester in the compo (Drugs and Pharmaceutical Science, Vol. 97. 235-239, sition to the total active agent in the composition is at least 4:1 1999), that skin can behave as a storage depot for absorbed fatty acid ester:active agent. Any composition as described materials. For example, askin storage depot for chemicals has above and below may be used in such methods. In specific been described by Vickers, Adv. Biol Skin. Vol. 12, 177-89 embodiments, the fatty acid ester is present in the composi (1972), to exist in the stratum corneum for topically applied tion in an amount ranging from 0.1% to 20% by weight of the lipophilic chemicals such as steroids. total weight of the pharmaceutical composition. 0029. However, as described herein, it has been found that 0017. In some embodiments, sustained release of the phar a storage depot in the dermis can be more effective than a maceutically active agent through the skin is observed at least storage depot in the stratum corneum, and can provide a better 24 hours after its administration. In other embodiments, Sus means to regulate the diffusion kinetics of active agents in the tained release of the pharmaceutically active agent through tissues and a better effective drug delivery over time. Indeed, the skin is observed at least 36 hours after its administration. the dermis constitutes the majority of the skin mass. It con US 2011/0098258 A1 Apr. 28, 2011

tains a dense blood and lymphatic vasculature, and it is the 4:1 fatty acid ester:active agent. In some embodiments, the site of drug absorption into the systemic circulation. The composition further comprises water, an alcohol and a fatty dermis, nevertheless, has rarely been targeted as a site for acid. In some embodiments, the composition even further administration or deposition of Substances, probably due to comprises a co-solvent, such as propylene glycol. Other con difficulty in controlling the layer of the skin in which the ventional components for transdermal pharmaceutical com active agent is actually retained. positions may also be included, as discussed in more detail 0030 Thus, described herein are transdermal pharmaceu below. tical compositions that exhibit advantageous properties and 0035. As used herein, the phrase “sustained delivery achieve advantageous results with regard to their drug deliv means that the compositions continue to deliver the active ery profiles. For example, embodiments of the compositions agent over a period of time of at least 12 hours, at least 24 described herein achieve systemic delivery of therapeutically hours, at least 36 hours, or at least 48 hours, including a period active agent(s) through the outer layers of the skin into the of time of at least 24 hours. For example, sustained delivery dermis, where a depot is formed from which the active agent compositions may continue to deliver the active agent after is delivered into the bloodstream over an extended period of the first 24 hours after application. In some embodiments, time. Such as over a period of time of at least 12 hours, at least Sustained delivery compositions described herein continue to 24 hours, at least 36 hours, or at least 48 hours. This can be deliver a therapeutically effective amount of the active agent observed, for example, when the active agent continues to be after the first 24 hours after application. Depending on the released into the bloodstream up to 24 hours, or longer, after composition and the active agent, this may constitute the skin wash. delivery after the first 24 hours of, for example, at least 5%, at 0031. In some embodiments, the compositions described least 10%, at least 15%, at least 20%, at least 25%, at least herein also advantageously achieve a high level of active 30%, or more, of the total amount of active agent delivered. agent delivery over a large range of active agent concentra Againdepending on the composition and the active agent, this tions. In addition, in some embodiments, the compositions may constitute the delivery after the first 24 hours of, for are formulated to promote reproducibility of absorption lev example, at least 2%, at least 3%, at least 4%, at least 5%, or els between different applications and between different more, of the total amount of active agent applied in the com patients. position. This can be observed, for example, when the active 0032. Thus, in accordance with some embodiments, there agent continues to be released into the bloodstream up to 24 are provided Sustained release transdermal pharmaceutical hours, or longer, after application. compositions for topical administration to a skin surface 0036. In some embodiments, sustained delivery composi comprising a pharmaceutically active agent and a fatty acid tions described herein continue to deliver active agent after ester, wherein the weight: weight ratio of the fatty acid ester the first 12, 24, 36, or 48 hours after application, at a level that in the composition to the active agent in the composition is at is greater than the amount delivered during the same time least 4:1 fatty acid ester:active agent. Such as ranging from 4:1 period by a comparable composition that does not include a to 20:1. In some embodiments, the composition further com fatty acid ester. As illustrated in Example 2, this can be prises water, an alcohol and a fatty acid. In some embodi observed by, for example, testing a composition as described ments, the composition even further comprises a co-solvent, herein and a comparable composition that does not include a Such as propylene glycol. Other conventional components for fatty acid ester (e.g., a composition that is identical except for transdermal pharmaceutical compositions may also be the absence of fatty acid ester) in an in vitro Franz cell assay, included, as discussed in more detail below. where the compositions are applied to a skin sample in the 0033. In particular, as discussed in more detail in the Franz cell, left for 24 hours, and then washed off, and then examples below, the compositions described herein relate to drug delivery through the skin after washing (e.g., after the the unexpected discovery that providing a fatty acid ester in at first 24 hours after application) is determined and compared. least a four-fold excess over the therapeutically active agent 0037. The compositions and methods are described in (on a W/w basis) results in a transdermal pharmaceutical more detail below, and illustrated in the examples. composition with advantageous properties, including Sus 0038. As used herein, and unless otherwise specified, “a tained release, consistent delivery profiles over a range of or “an means “one or more. active agent concentrations, and application-to-application 0039. The term “about' and the use of ranges in general, and patient-to-patient reproducibility. While not wanting to whether or not qualified by the term about, means that the be bound by any theory, it is believed that this high fatty acid number comprehended is not limited to the exact number set ester to active agent ratio facilitates partitioning of the active forth herein, and is intended to refer to ranges substantially agent into the dermis and formation of a depot within the within the quoted range while not departing from the scope of dermis, resulting in dermal retention of the active agent fol the invention. As used herein, “about will be understood by lowed by sustained release into the bloodstream. Hence, the persons of ordinary skill in the art and will vary to some extent compositions and methods described herein also provide a on the context in which it is used. If there are uses of the term method for increasing dermal retention of an active agent, and which are not clear to persons of ordinary skill in the art given achieving Sustained release delivery. the context in which it is used, “about will meanup to plus or 0034. Thus, in accordance with some embodiments, there minus 10% of the particular term. is provided a method for providing a Sustained release of a 0040 Pharmaceutical Compositions pharmaceutically active agent through the skin of a subject, 0041 As noted above, described herein are compositions comprising topically administering to the skin of the Subject comprising a therapeutically effective amount of a therapeu a pharmaceutical composition comprising a therapeutically tically active agent and a fatty acid ester, wherein the weight: effective amount of the active agent and a fatty acid ester, weight ratio of the fatty acid ester in the composition to the wherein the weight: weight ratio of the fatty acid ester in the active agent in the composition is at least 4:1 fatty acid ester: composition to the active agent in the composition is at least active agent, such as ranging from 4:1 to 20:1. In particular US 2011/0098258 A1 Apr. 28, 2011 embodiments, the compositions comprise a pharmaceutically mones include glucocorticoids, thyroid hormone, calciferol, active agent, a fatty acid ester, water, an alcohol and a fatty pregnenolone, aldosterone, cortisol, and derivatives thereof. acid. In further particular embodiments, the composition fur Suitable steroid hormones especially include the sexual hor ther comprises a co-solvent, Such as propylene glycol. mones having estrogenic, progestational, androgenic, orana 0042. In specific embodiments, the composition com bolic effects, such as estrogen, estradiol and their esters, e.g., prises about 2% fatty acid (such as oleic acid), about 2% fatty the Valerate, benzoate, or undecylate, ethinylestradiol, etc.; acid ester (such as ethyl oleate), and about 5% co-solvent progestogens, such as norethisterone acetate, levonorgestrel, (such as propylene glycol). In further specific embodiments, chlormadinone acetate, cyproterone acetate, desogestrel, or the composition comprises 2% fatty acid (such as oleic acid), gestodene, etc.; androgens. Such as testosterone and its esters 2% fatty acid ester (such as ethyl oleate), and 5% co-solvent (propionate, undecylate, etc.), etc.; anabolics, such as meth (such as propylene glycol). In other specific embodiments, androstenolone, nandrolone and its esters. the composition comprises about 0.3% fatty acid (such as 0049 Estrogens oleic acid), about 0.3% fatty acid ester (such as ethyl oleate), 0050. In specific embodiments, the one or more estrogen and about 0.75% co-solvent (such as propylene glycol). In (s) are selected from the group consisting of natural oestro further specific embodiments, the composition comprises gens, such as 17B-Oestradiol, oestrone, equine conjugated 0.3% fatty acid (such as oleic acid), 0.3% fatty acid ester oestrogens, estriol and phytoestrogens; semi-natural oestro (such as ethyl oleate), and 0.75% co-solvent (such as propy gens, such as oestradiol Valerate; or synthetic oestrogens, lene glycol). As noted above, as used herein, the term “about Such as ethinyl-estradiol. embraces plus or minus 10% of the listed amounts. 0051. In some embodiments, a pharmaceutical composi 0043. In some embodiments, the composition comprises tion is provided for topical administration to a skin Surface the specified components. In some embodiments, the compo comprising water, at least one therapeutically active agent sition consists of the specified components. In other embodi selected from the estrogens, an alcohol, and a fatty acid ester. ments, the composition consists essentially of the specified In some embodiments, a pharmaceutical composition is pro components. As used herein, "consists essentially of the vided for topical administration to a skin Surface comprising specified components means that the composition includes at water, at least one therapeutically active agent being estradiol. least the specified components, and may also include other analcohol, and a fatty acid ester. In particular embodiments of components that do not materially affect the basic and novel Such compositions when the active agent is estradiol, the characteristics of the invention. composition does not further comprise the combination of 0044 Specific components of the compositions are progesterone, propylene glycol, oleic acid, ethyl oleate, etha described in detail below. nol, hydroxypropylcellulose and purified water. 0045 Active Agents 0.052 Anti-Estrogens 0046. The compositions described herein include at least 0053 Anti-estrogens are a class of pharmaceutically one therapeutically active agent. In some embodiments, the active agents now referred to as Selective Estrogen Receptors active agent is a drug molecule of generally hydrophobic Modulators (SERMs), which were generally understood to be nature, with a small size, such as a molecular weight below compounds capable of blocking the effect of estradiol with 500 Dalton. In some embodiments, the active agent is out displaying any estrogenic activity of their own. Such a selected from steroids, including hormones and sex hor description is now known to be incomplete, however. The mones. The term “sex hormone' refers to natural or synthetic term SERM has been coined to describe compounds that, in steroid hormones that interact with vertebrate androgen or contrast to pure estrogen agonists or antagonists, have a estrogen receptors, such as estrogens, anti-Oestrogens (or mixed and selective pattern of estrogen agonist-antagonist SERMs), androgens, anti-androgens, progestins, and mix activity, which largely depends on the targeted tissue. The tures thereof. When the composition comprises more than pharmacological goal of these drugs is to produce estrogenic one steroid, the weight: weight ratio of the fatty acid ester in actions in those tissues where these actions are beneficial the composition to the total amount of steroid in the compo (such as bone, brain, liver) and to have either no activity or sition is at least 4:1 fatty acid ester:Steroids, such as ranging antagonistic activity in tissues such as breast and from 4:1 to 20:1. endometrium, where estrogenic actions (cellular prolifera 0047. In some embodiments where the composition com tion) might be deleterious. prises one or more steroids and one or more other therapeu 0054. In specific embodiments, the anti-estrogens tically active agents, the weight:weight ratio of the fatty acid (SERMs) are selected from the group consisting of endox ester in the composition to the total amount of active agents in ifen, droloxifene, clomifene, raloxifene, tamoxifen, 4-OH the composition is at least 4:1 fatty acid esteractive agents. In tamoxifen, toremifene, danazol, and pharmaceutically other embodiments, the weight: weight ratio of the fatty acid acceptable salts thereof. In a more particular embodiment, a ester in the composition to the total amount of active agents in pharmaceutical composition is provided for topical adminis the composition is less than 4:1 fatty acid ester:active agent, tration to a skin Surface comprising water, at least one thera although the weight:weight ratio of the fatty acid ester in the peutically active agent selected from the anti-Oestrogens composition to the total amount of steroid in the composition (SERMs) selected from the group consisting of clomifene, is at least 4:1 fatty acid ester:steroid. raloxifene, droloxifene, endoxifen or the pharmaceutically 0048 For example, steroid hormones suitable for use in acceptable salts thereof, an alcohol, and a fatty acid ester. the compositions described herein include the numerous 0055. In a particular embodiment, a pharmaceutical com natural and synthetic steroid hormones, including androgens, position is provided for topical administration to a skin Sur estrogens, and progestagens and derivatives thereof. Such as face comprising water, at least one therapeutically active dehydroepiandrosterone (DHEA), androstenedione, andros agent selected from the anti-estrogens (SERMs), an alcohol, tenediol, dihydrotestosterone, testosterone, progesterone, and a fatty acid ester. In some particular embodiments of such progestins, oestriol, oestradiol. Other suitable steroid hor compositions, when the active agent is tamoxifen, the fatty US 2011/0098258 A1 Apr. 28, 2011 acid ester is not isopropyl myristate. In other particular face comprising water, at least one therapeutically active embodiments of Such compositions, when the active agent is agent selected from the progestins, an alcohol, and a fatty acid tamoxifen, the composition further comprises a fatty acid. In ester. In particular embodiments of such compositions, when yet other particular embodiments of such compositions, when the active agent is progesterone, the composition does not the active agent is 4-OH tamoxifen, the fatty acid ester is not further comprise the combination of estradiol, propylene gly isopropyl myristate. In other particular embodiments of Such col, oleic acid, ethyl oleate, ethanol, hydroxypropylcellulose compositions, when the active agent is 4-OH tamoxifen, the and purified water. composition further comprises a fatty acid. 0056 Androgens 0068. In particular embodiments, the therapeutically 0057. In some embodiments, androgens may be selected active agent in the pharmaceutical composition is a progestin, from the group consisting of the natural androgen, testoster an estrogen or a combination of the two. one, and its semi-natural or synthetic derivatives, for instance 0069. As noted above, when the composition comprises methyltestosterone; physiological precursors of testosterone more than one steroid, the weight:weight ratio of the fatty such as dehydroepiandrosterone or DHEA, or alternatively acid ester in the composition to the total amount of steroid prasterone and its derivatives, for instance DHEA sulphate, active agent in the composition is at least 4:1 fatty acid ester: A-4-androstenedione and its derivatives; testosterone active agent, Such as ranging from 4:1 to 20:1. metabolites, for instance dihydrotestosterone (DHT) 0070 The amount of therapeutically active agent present obtained after the enzymatic action of 5-O-reductases; or in the composition generally will be influenced by the dosage Substances with an androgenic-type effect, Such as tibolone. to be delivered for therapeutic effect and formulary consid 0058. In a particular embodiment, a pharmaceutical com erations. The compositions generally include a therapeuti position is provided for topical administration to a skin Sur face comprising water, at least an active agent selected from cally effective amount of active agent. As used herein, the the androgens, an alcohol, and a fatty acid ester. In particular phrase “therapeutically effective amount’ means an amount embodiments of Such compositions, when the active agent is (dosage) that achieves in a Subject the specific pharmacologi testosterone or dihydrotestosterone (DHT), the composition cal response for which the drug is administered. It is empha also comprises a fatty acid as a penetration enhancer. sized that a “therapeutically effective amount of a drug that 0059 Anti-Androgens is administered to a particular subject in a particular instance 0060. In some embodiments, the anti-androgens are may not always be effective in treating the target conditions/ Selected from the group consisting of steroidal compounds diseases, even though such dosage is deemed to be a thera Such as cyproterone acetate and medroxyprogesterone, or peutically effective amount by those of skill in the art. Those non-steroidal compounds such as flutamide, nilutamide or skilled in the art will recognize that the “therapeutically effec bicalutamide. tive amount may vary from patient to patient, or from con 0061. In a particular embodiment, a pharmaceutical com dition to condition, and can determine a “therapeutically position is provided for topical administration to a skin Sur effective amount for a given patient/condition by routine face comprising water, at least one active agent selected from CaS. the anti-androgens, an alcohol, and a fatty acid ester. 0071. The therapeutically active agent is advantageously 0062 Progestins present in the composition in an amount ranging from about 0063. In some embodiments, the progestin(s) used in the 0.01% to about 5%, or from 0.01% to 5%, including from pharmaceutical compositions described herein may be about 0.02% to about 3%, or from 0.02% to 3%, such as from selected from the group consisting of natural progestins, about 0.03% to about 2%, or from 0.03% to 2%, including progesterone or its derivatives of ester type, and synthetic from about 0.05% to about 0.5%, or from 0.05% to 0.5%, progestins of type 1, 2 or 3. such as from about 0.2% to about 0.4%, or from 0.2% to 0064. The first group comprises molecules similar to 0.4%, these percentages being expressed by weight, relative progesterone or the synthetic progestins 1 (SP1) (pregnanes), to the total weight of the pharmaceutical composition. for example the progesterone isomer (retroprogesterone), 0072 According to one embodiment, when the active medrogesterone, and norprogesterone derivatives (demege agent comprises a progestin, the progestin content ranges stone or promegestone). from about 0.01% to about 5%, including from about 0.05% 0065. The second group comprises 17a-hydroxy-progest to about 3%, such as from about 0.1% to about 1%, these erone derivatives or synthetic progestins 2 (SP2) (pregnanes), percentages being expressed by weight, relative to the total for example cyproterone acetate and medroxyprogesterone weight of the pharmaceutical composition. Thus, in some acetate. embodiments, the progestin content may range from 0.01% to 0066. The third group comprises norsteroids or synthetic 5%, including from 0.05% to 3% such as from 0.1% to 1%, progestins 3 (SP3), (estranes or nor-androstanes). These are 0073. According to another embodiment, when the active 19-nortestosterone derivatives, for example norethindrone. agent comprises an estrogen, the estrogen content ranges This group also comprises molecules of gonane type, which from about 0.01% to about 5%, including from about 0.02% are derived from these nor-androstanes or estranes and have a to about 3%, such as from about 0.03% to about 2%, includ methyl group at C18 and an ethyl group at C13. Examples that ing from about 0.05% to about 0.50%, such as from about may be mentioned include norgestimate, desogestrel(3-ke 0.20% to about 0.40%, these percentages being expressed by todesogestrel) or gestodene. Tibolone, which has both weight, relative to the total weight of the pharmaceutical progestin and androgenic activity, may also advantageously composition. Thus, in Some embodiments, the estrogen con be selected in the pharmaceutical composition described tent may range from 0.01% to 5%, including from 0.02% to herein. 3%, such as from 0.03% to 2%, including from 0.05% to 0067. In a particular embodiment, a pharmaceutical com 0.50%, such as from 0.20% to 0.40%, including from about position is provided for topical administration to a skin Sur O.30% to 0.40%. US 2011/0098258 A1 Apr. 28, 2011

0074. In one embodiment, when the active agent com content may range from 0.01% to 5%, including from 0.05% prises an estrogen, the estrogen content will range from about to 2.4%, such as from 0.1% to 2.2%, O.30% to 0.40%. I0082 Fatty Acid 0075 Fatty Acid Ester I0083. In some embodiments, the composition may com 0076. The compositions described hereincomprise at least prise at least one fatty acid that can either be saturated or one fatty acid ester. unsaturated, such as a fatty acid penetration enhancer. Exem 0077. The fatty acid esters suitable for use herein include plary fatty acids suitable for use include long-chain aliphatic long-chain aliphatic fatty acid esters containing from 8 to 22 fatty acids containing from 8 to 22 carbonatoms, such as from carbon atoms, such as from 12 to 20 carbon atoms. The fatty 10 to 18 carbon atoms. The fatty acids may be selected, in a acid esters may be those that would result from the reaction of non-limiting manner, from the group consisting of capric acid analcohol with a fatty acid selected, in a non-limiting manner, (10:0), lauric acid (12:0), myristic acid (14:0), palmitic acid from the group consisting of capric acid (10:0), lauric acid (16:0), stearic acid (18:0); oleic acid (18:1), isostearic acid (12:0), myristic acid (14:0), palmitic acid (16:0), stearic acid (18:0), palmitoleic acid (16:1), linoleic acid (18:2) and lino (18:0), oleic acid (18:1), isostearic acid (18:0), palmitoleic lenic acid (18:3). In a particular embodiment, the fatty acid is acid (16:1), linoleic acid (18:2) and linolenic acid (18:3). oleic acid. 0078 Thus, for example, the fatty acid ester can optionally I0084. In a particular embodiment, the fatty acid formu be selected from the group consisting of ethyl oleate, isopro lated in the composition corresponds to the fatty acid ester pyl oleate, isopropyl myristate, isopropyl isostearate, isopro also formulated in the composition, such as a composition pyl palmitate, ethyl octanoate, ethyl dodecanoate, ethyl comprising ethyl oleate and oleic acid. Thus, in a particular linoleate, ethyl palmitoleate, ethyl isostearate and ethyllino embodiment, the composition comprises both oleic acid and lenate. In a particular embodiment, the fatty acid ester is an at least one of its corresponding esters. In other embodiments, ester that would result from the reaction of an alcohol with the fatty acid formulated in the composition does not corre oleic acid. spond to the fatty acid ester also formulated in the composi 0079. In one embodiment, the fatty acid ester is an ester tion. that would result from the reaction of the fatty acid formu I0085. The fatty acid content in the pharmaceutical com lated in the composition with an alcohol. In other embodi position described herein will advantageously range from ments, the fatty acid ester is not an ester that would result from about 0.01% to about 5%, including from about 0.05% to the reaction of the fatty acid formulated in the composition about 3.5%, such as from about 1% to about 3%, these per with an alcohol. In one embodiment, the fatty acid ester is not centages being expressed by weight, relative to the total the ester that would result from the reaction of the fatty acid weight of the pharmaceutical composition. Thus, in some formulated in the composition with the alcohol formulated in embodiments, the fatty acid content may range from 0.01% to the composition. For example, in the context of the disclosed 5%, including from 0.05% to 3.5%, such as from 1% to 3%, compositions, the advantageous results discussed herein, I0086 Alcohol As noted above, the compositions such as sustained delivery believed to be due to formation of described herein comprise at least one alcohol. As used herein a depot in the dermis, may be observed without regard to the term “alcohol refers to an organic molecule containing at whether the fatty acid ester formulated in the composition least one carbon atom and only one alcohol group —OH corresponds to any fatty acid also formulated in the compo (monoalcohol). sition. I0087 Exemplary alcohols are C2-C6 alcohols and can 0080. As noted above, that fatty acid ester is present in the include C2-C4 alcohols, such as ethanol, n-propanol, isopro composition in at least a four-fold excess over the therapeu panol, n-butanol, isobutanol, tert-butanol, or mixtures tically active agent (on a W/w basis), i.e., the weightweight thereof. Exemplary, non-limiting alcohols Suitable for use in ratio of the fatty acid ester present in the composition to the the compositions are ethanol and isopropanol. active agent present in the composition is at least 4:1 fatty acid I0088. The presence of such an alcohol may also accelerate ester:active agent. Thus, in Some embodiments, the weight: drying of the composition onto the skin. For that reason, weight ratio of the fatty acid ester:active agent ranges from alcohols may be chosen that have a boiling point in the range 4:1 to 20:1. In another embodiment, the weight:weight ratio of about 70 to about 130°C., including in the range of about of the fatty acid ester present in the composition to the active 75 to about 85°C. agent present in the composition ranges from 4:1 to 15:1; in I0089. Typically, the alcohol will be used in an amount further embodiments, the ranges is from 5:1 to 10:1, or from ranging from about 10% to about 90%, including from about 5:1 to 7:1. Within these parameters, the fatty acid ester con 20% to about 80%, such as from about 45% to about 75%, tent in the pharmaceutical composition may range from about these percentages being expressed by weight, relative to the 0.1% to about 20% by weight, such as from about 0.2% to total weight of the pharmaceutical composition. Thus, in about 10% by weight, including from about 0.5% to about 5% Some embodiments, the alcohol may be present in an amount by weight, all based on the total weight of the pharmaceutical ranging from 10% to 90%, including from 20% to 80%, such composition. Thus, the compositions may comprise fatty acid as from 45% to 75%. ester in an amount from 0.1% to 20% by weight, such as from 0090 Co-Solvent 0.2% to 10% by weight, including from 0.5% to 5% by 0091. The pharmaceutical composition described herein weight. may also comprise a co-solvent. Co-solvents Suitable for use 0081. In particular embodiments, the fatty acid ester con in pharmaceutical compositions are known in the art, Such as tent in the pharmaceutical composition may range from about polyols or polyglycols. In some embodiments, one or more 0.01% to about 5%, including from about 0.05% to about co-solvents are selected from the group consisting of glyc 2.4%, such as from about 0.1% to about 2.2%, these percent erol, propylene glycol and polyethylene glycol. ages being expressed by weight, relative to the total weight of 0092. The co-solvent may be present in the composition in the pharmaceutical composition. Thus, the fatty acid ester an amount ranging from about 0.01% to about 7%, including US 2011/0098258 A1 Apr. 28, 2011

from about 3% to about 7%, such as from about 4% to about 0102. As used herein “moisturizer specifies an agent that 6%, these percentages being expressed by weight, relative to hydrates the skin. Moisturizers suitable for use in pharmaceu the total weight of the pharmaceutical composition. Thus, in tical compositions are known in the art. Moisturizers can be Some embodiments, the co-solvent may be present in an used either alone or in combination, e.g., a combination of amount ranging from 0.01% to 7%, including from 3% to 7%, two or three (or more) different moisturizers can be used. In such as from 4% to 6%. Some embodiments, moisturizers are selected from emol 0093. The co-solvent generally increases the solubility of lients and/or humectants. the therapeutically active agent(s) and in particular may con 0103) As used herein, “emollients’ specify substances that tribute to maintain in solution the therapeutically active agent soften the skin and tend to improve moisturization of the skin. remaining on the skin Surface once the alcohol has dried. For Emollients Suitable for use in pharmaceutical compositions that reason, co-solvents may be selected that have a boiling are well known in the art, and include mineral oil, petrolatum, point in the range of about 150° C. to about 300° C., such as polydecene, isohexadecane, fatty acids and alcohols having in the range of about 150° C. to about 200° C. from 10 to 30 carbon atoms; pelargonic, lauric, myristic, 0094 Gelling Agents palmitic, Stearic, isostearic, hydroxyStearic, oleic, linoleic, 0095. The compositions may optionally comprise at least ricinoleic, arachidic, behenic, and euricic acids and alcohols; one gelling agent. triglyceride esters, castor oil, cocoa butter, safflower oil, Sun flower oil, jojoba oil, cottonseed oil, corn oil, olive oil, cod 0096. As used herein, the term “gelling agent” specifies a liver oil, almond oil, avocado oil, palm oil, Sesame oil, compound, optionally of polymeric nature, having the capac squalene, Kikui oil, soybean oil, acetoglyceride esters, ity to form a gel when contacted with a specific solvent, e.g., ethoxylated glycerides, ethoxylated glyceryl monostearate, water. Gelling agents (e.g., thickeners) Suitable for use in alkyl esters offatty acids having 10 to 20 carbonatoms, hexyl pharmaceutical compositions are known in the art. Gelling laurate, isohexyllaurate, isohexyl palmitate, isopropyl palmi agents may act to increase the viscosity of the pharmaceutical tate, decyl oleate, isodecyl oleate, hexadecyl Stearate, decyl compositions. For example, a gelling agent may provide the Stearate, diisopropyl adipate, diisohexyl adipate, diisopropyl composition with Sufficient viscosity to allow easy applica sebacate, laurly lactate, myristyl lactate, acetyl lactate; alk tion of the composition onto the skin. Additionally or alter enyl esters of fatty acids having 10 to 20 carbon atoms, oleyl natively, gelling agents may act as solubilizing agents. myristate, oleyl Stearate, oleyl oleate, fatty acid esters of 0097 Examples of gelling agents include anionic poly ethoxylated fatty alcohols, polyhydric alcohol esters, ethyl merS Such as acrylic acid based polymers (including poly ene glycol mono and di-fatty acid esters, diethylene glycol acrylic acid polymers, e.g. Carbopol R by Noveon, Ohio), mono- and di-fatty acid esters, polyethylene glycol, wax cellulose derivatives, poloxamers and poloxamines, more esters, beeswax, spermaceti, myristyl myristate, Stearyl Stear precisely, Carbomers which are acrylic acid-based polymers, ate, silicone oils, dimethicones, cyclomethicones. In some e.g. Carbopol.R. 980 or 940,981 or 941, 1342 or 1382, 5984, embodiments, the composition comprises one or more emol 934 or 934P (Carbopol R are usually polymers of acrylic acid lients that are liquid at room temperature. crosslinked with allyl sucrose orallylpentaerythritol), Ultrez, 0104. As used herein “humectants' specifies hygroscopic Pemulen TR1(R) or TR2R) commercialized by Lubrizol (high substances that absorb water from the air. Exemplary humec molecular weight copolymer of acrylic acid and C10-C30 tants Suitable for use include glycerine, propylene glycol, alkyl acrylate crosslinked with allyl pentaerythritol), Syntha glyceryl triacetate, a polyol, Sorbitol, maltitol, a polymeric len CR, etc.; cellulose derivatives such as carboxymethylcel luloses, hydroxypropylcelluloses (Klucel(R), for example polyol, polydextrose, quillaia, lactic acid, and urea. Klucel HFR, or Klucel HPC(R) sold by Hercules Incorpo 0105 Exemplary moisturizers suitable for use may com rated), hydroxyethylcelluloses, ethylcelluloses, hydroxym prise amines, alcohols, glycols, amides, Sulfoxides, and pyr ethylcelluloses, hydroxypropylmethylcelluloses, and the rolidones. In one aspect, the moisturizer is selected from the like, and mixtures thereof; poloxamers or polyethylene group consisting of lactic acid, glycerine, propylene glycol, polypropylene copolymers such as Lutrol(R) grade 68 or 127, and urea. poloxamines and other gelling agents such as chitosan, dex 0106. In one embodiment, the moisturizer is used in an tran, pectins, and natural gums. Any one or more of these amount ranging from about 0.01% to about 30% by weight, gelling agents may be used alone or in combination in the including from about 0.05% to about 20% by weight, such as pharmaceutical compositions described herein. In one aspect, from about 0.1% to about 10% by weight, including from the gelling agent is selected from the group consisting of about 0.5% to about 5% by weight, these percentages being expressed by weight, relative to the total weight of the phar polyacrylic acids, cellulosics, and mixtures thereof. maceutical composition. Thus, for example, a moisturizer 0098. In one embodiment, the compositions described may be used in an amount ranging from 0.01% to 30% by herein comprise Pemulen R as a gelling agent. weight, including from 0.05% to 20% by weight, such as from 0099 Typically, the gelling agent will be used in an 0.1% to 10% by weight, including from 0.5% to 5% by amount ranging from about 0.05% to about 5% by weight, weight. including about 0.1% to about 3%, such as from about 1.5% 0107. In one embodiment, the composition comprises to about 2.5% by weight, these percentages being expressed glycerin in an amount ranging from about 0.01% to about by weight, relative to the total weight of the pharmaceutical 30% by weight, including from about 0.05% to about 20% by composition. Thus, the gelling agent may be present in an weight, such as from about 0.1% to about 10% by weight, amount ranging from 0.05% to 5% by weight, including 0.1% including from about 0.5% to about 5% by weight, these to 3%, such as from 1.5% to 2.5% by weight. percentages being expressed by weight, relative to the total 01.00 Moisturizers weight of the pharmaceutical composition. Thus, in some 0101 The compositions may optionally comprise at least embodiments, a composition may comprise glycerin in an one moisturizer. amount ranging from 0.01% to 30% by weight, including US 2011/0098258 A1 Apr. 28, 2011

from 0.05% to 20% by weight, such as from 0.1% to 10% by mine, sodium hydroxide, ammonium hydroxide, potassium weight, including from 0.5% to 5% by weight. hydroxide, arginine, aminomethylpropanol or tromethamine, 0108 Aqueous Vehicle and mixtures thereof. Where the pH of the pharmaceutical 0109 As noted above, the composition described herein composition is not optimized for transdermal administration, comprises an aqueous vehicle, and thus includes water. Aque e.g., where the gelling agent comprises at least one acrylic ous vehicles Suitable for pharmaceutical compositions are acid-based polymer resulting in a pH more acidic than desired known in the art. for the final product, the use of a base may contribute to the 0110. According to one embodiment, the aqueous vehicle neutralization of the pharmaceutical composition. Further comprises, besides water, ingredients useful in adjusting the more, the use of the base (neutralizer) may improve or opti pH, for instance at least one buffering agent, which advanta mize Swelling of the polymer chains during the neutralization geously makes it possible to maintain the pH of the compo of the charges and the formation of polymer salts. In embodi sition between about 4 and about 10, such as between about 5 ments where the gelling agent comprises an acrylic acid and about 9, or between about 6 and about 8, including from based polymer, the base may comprise triethanolamine. The 4 to 10, from 5 to 9 and from 6 to 8. use of a base also may improve or optimize viscosity. 0111. According to another embodiment of the pharma I0121 The skilled person will know how to choose a suit ceutical composition, the buffers are selected from the group able amount of base for use in the composition, and may consisting of: select the base based on the nature of the gelling agent present 0112 basifying or basic buffers such as a phosphate therein, and the alcohol content of the composition. For buffer (for example dibasic or monobasic sodium phos example, with carbomers and/or a high alcohol content, phate), a citrate buffer (for example sodium citrate or tromethamine and/or NaOH may be selected as a base, in potassium citrate), sodium carbonate, sodium bicarbon amounts chosen so as to reach the desired final pH in the ate, including a mixture of sodium carbonate and sodium composition. bicarbonate, or 0.122 Further Optional Components 0113 neutral buffers such as a Tris buffer (for example I0123. The pharmaceutical compositions described herein tris maleate), or a phosphate buffer. optionally may comprise other usual pharmaceutical addi 0114. In one embodiment, the compositions comprise a tives, including salt(s), stabilizer(s), antimicrobial(s) such as mixture of sodium carbonate and sodium bicarbonate. paraben compounds, fragrance(s), and/or propellant(s). 0115 The buffer can be introduced in the composition 0.124. In some embodiments, it may for example be advan either directly, for example added in a powdery form, or tageous to include a stabilizer such as butylated hydroxy diluted in water, for example to a concentration ranging from anysol (BHA), butylated hydroxytoluene (BHT) and ascorbic 1 to 500 mM. Thus, additionally or alternatively, the liquid acid. BHA, however, may color the compositions in yellow. buffer solution can be introduced in the composition. Therefore, in another embodiment, the composition does not 0116. The skilled artisan would understand how to adjust comprise BHA. the amount of buffer to obtain the desired buffering effect, 0.125 Depending on the nature of the selected ingredients, depending on the chemical nature of the buffer used, its form it may be advantageous to include a Surfactant. Surfactants (either powdery or diluted in water) and the starting and Suitable for use in pharmaceutical compositions are known in desired pH of the composition. the art, and the skilled person can select Suitable Surfactants 0117. Without being limited to these values, it can reason for use in the compositions described herein, such as Surfac ably be estimated that when the buffers used in the composi tants that are dermatologically and/or cosmetically accept tion are a mixture of sodium carbonate and sodium bicarbon able. Examples thereof include non-ionic Surfactants, for ate introduced in a powdery form (see example 8 of the example: present application), sodium carbonate can be introduced in 0.126 esters, such as: an amount ranging from about 0.01 to 0.1%, and sodium I0127 esters of polyethyleneglycol with fatty acids, bicarbonate can be introduced in an amount ranging from about 0.001 to 0.01%, these percentages being expressed by including Labrasol R, which is a mixture of mono-, di weight, relative to the total weight of the pharmaceutical and triglycerides and of mono- and diesters of poly composition. ethyleneglycol with fatty acids; 0118 Without being limited to these values, it can reason I0128 esters of saccharose with fatty acids, such as ably be estimated that when the buffer used in the composi sucrose laurate with HLB16; sucrose palmitate with tion is a 60 mM solution of carbonate buffer having a HLB 16; pH=10.7 (see examples 1 to 3 of the present application), the I0129 esters of sorbitanne polyoxyethylene, such as 60 mM buffer solution can be introduced in an amount rang Tween(R) compounds including Tween(R) 20, 60 and/or ing from about 1% to about 80%, including from about 5% to 80; about 70%, such as from about 10% to about 50%, these 0.130 alkylene oxide copolymers, such as copolymers percentages being expressed by weight, relative to the total of ethylene oxide and propylene oxide, e.g. Pluronics(R). weight of the pharmaceutical composition. I0131 Further examples include anionic surfactants such 0119) However, the amount of buffer in the composition as SDS (sodium dodecyl sulphate), and the like and cationic can further vary depending on the composition of the formula Surfactants such as cetrimide(alkyltrimethylammonium bro in which it is introduced, in accordance with standard buff mide) and the like. ering techniques. I0132 Typically, surfactants will be used in the composi 0120 In another aspect, the pharmaceutical compositions tions in an amount ranging from about 0.01% to about 5% by described herein further comprises a base. Advantageously, weight, including about 0.05% to about 3% by weight, these the base is, for example, pharmaceutically acceptable, and is percentages being expressed by weight, relative to the total typically selected from the group consisting of triethanola weight of the pharmaceutical composition. Thus, in some US 2011/0098258 A1 Apr. 28, 2011 embodiments, a surfactant may be used in the compositions in 0154 When the active agent is an antiestrogen (SERM), an amount ranging from 0.01% to 5% by weight, including compositions described herein are useful for treating a 0.05% to 3% by weight. patient Suffering from or at risk of developing a breast 0133. The pharmaceutical composition described herein disorder Such as: may be in the form of a solution, a gel, a cream, a lotion, a 0155 conditions involving dense breast tissue, such milk, an ointment, an aerosol or a patch. as high density breast tissue that is a predictor of 0134. In one embodiment, the composition is in the form breast cancer risk and/or that compromises mammo of a gel or a solution. graphic sensitivity; 0135 Exemplary Composition and Uses 0156 benign breast diseases. Such as adenosis, cysts, 0.136 Exemplary, non-limiting compositions are provided duct ectasia, fibroadenoma, fibrosis, hyperplasia, below. As mentioned above, percentages (%) refer to amounts metaplasia and other fibrocystic changes; by weight based upon the total weight of the composition 0157 gynecomastia; (w/w). The sum of the different components of the composi 0158 breast cancer, including non-invasive breast tion adds up to 100% (w/w) of the total composition. Cancer, 0137 In one aspect, a pharmaceutical composition is pro 0159 malignant melanoma; vided for topical administration to a skin surface wherein the 0160 mastalgia; composition comprises: 0.161 localized cancer and/or tumours such as lung I0138 (i) 0.01 to 2.5% (w/w) of a pharmaceutically tumours; and active agent comprising one or more steroids, 0162 other therapies involving the systemic admin I0139 (ii) 10 to 90% (w/w) of at least one C2-C6 istration of an antiestrogen. monoalcohol, such as ethanol or isopropanol, 0.163 When the active agent is an estrogen such as (O140 (iii) 0.04 to 10% (w/w) of a fatty acid ester, estradiol, an antiestrogen (SERM), an androgen such as 0141 (iv) 0 to 10% (w/w) of a fatty acid testosterone or DHT, compositions described herein are 0.142 (v) 0 to 5% (w/w) of at least one gelling agent, useful for treating a bone-related disorder Such as 0143 (vi) q.s.f. 100% (w/w) water, osteoporosis, menopause-associated osteoporosis, glu 0144 wherein the weight: weight ratio of the fatty acid cocorticoid-induced osteoporosis, Paget's disease, ester in the composition to the total active agent in said abnormal bone resorption, bone cancer, bone loss (gen composition is at least 4:1 fatty acid ester:active agent. eralized bone loss and/or localized bone loss), bone (0145. In one embodiment, a pharmaceutical composition metastasis (with or without hypercalcemia), multiple is provided for topical administration to a skin Surface myeloma and other conditions that feature bone fragil wherein the composition comprises: ity. 0146 (i) 0.01 to 1.25% (w/w), such as 0.30 to 0.50% 0164. When the active agent is an estrogen such as (w/w), of a pharmaceutically active agent chosen from estradiol, compositions described herein are useful for estrogens, such as estradiol, 0.165 the prevention of cardiovascular diseases or 0147 (ii) 20 to 80% (w/w) of at least one C2-C6 improvement of cognitive functions; monoalcohol, such as ethanol or isopropanol, 0166 managing symptoms of menopause, such as 0148 (iii) 0.04 to 5% (w/w) of a fatty acid ester, such as hot flashes, night Sweats, sleeping problems (insom ethyl oleate nia), fatigue, vaginal dryness and itching and burning, 0149 (iv) 0.01 to 5% (w/w) of a fatty acid, such as oleic loss of sexual desire, irregular periods, bladder prob acid, lems, and mood Swings; (O150 (v) 0.05% to 5% (w/w) of at least one gelling 0.167 prostate cancer; and agent, such as a high molecular weight copolymer of 0168 other therapies involving the systemic admin acrylic acid and C10-C30 alkyl acrylate crosslinked istration of an estrogen. with allyl pentaerythritol, for example PemulenRTR-1, 0169. When the active agent is a progestogen such as 015.1 (vi) q.s.f. 100% (w/w) water, progesterone, compositions described herein are useful 0152 wherein the weight: weight ratio of the fatty acid for treating ester in the composition to the total active agent in said 0170 begnin breast disease, mastodynia, mastopa composition is at least 4:1 fatty acid ester:active agent, Such thy, cyclical mastalgia, and to prevent cysts and beg as ranging from 4: 1 and 7:1. nin tumor relapse. 0153. Depending on the active agent used, the pharmaceu 0171 pre-menstrual syndrome, menstrual irregulari tical compositions described herein can be useful for various ties due to ovulation disorders oranovulation, benign treatments. For example, the compositions can be used in any mastopathy, premenopause, adjunctive use with methods where the delivery of a pharmaceutically active oestrogen in post-menopausal women, prevention of agent is desired, and may be particularly useful where sus endometrial hyperplasia in non-hysterectomized tained, systemic delivery of the pharmaceutically active agent postmenopausal women who are receiving estrogen is desired. When the composition comprises one or more therapy, infertility due to luteal phase defect, threat steroids, it can be used in any method where the delivery of ened abortion, and threatened preterm delivery, the steroid(s) is desired, and may be particularly useful where progesterone Support during ovarian insufficiency or sustained, systemic delivery of the steroid(s) is desired. For complete ovarian failure, in women lacking ovarian example, the compositions can be used in methods to treat a function (oocyte donation), for luteal phase Support patient Suffering from or at risk of developing any condition during in vitro fertilisation cycles, for luteal phase that may be treated, ameliorated or prevented by the systemic Support during spontaneous or induced cycles, in pri administration of one or more Steroids, Exemplary, non-lim mary or secondary infertility or subfertility in particu iting therapeutic methods include: lar due to dySovulation; and US 2011/0098258 A1 Apr. 28, 2011 10

0172 other therapies involving the systemic admin prepare the pharmaceutical compositions by any Suitable istration of a progestogen. means, based on common general knowledge. For example, 0173 When the active agent is an androgen such as the pharmaceutically active agent(s) can be dissolved in the Testosterone or DHT, compositions described herein are alcohol and mixed with the aqueous vehicle (e.g., water and useful for treating: other optional components discussed above) and co-solvent, 0.174 hypogonadism; if being used, followed by addition of the other excipients, 0.175 depressive disorder, type-2 diabetes, increas Such as the moisturizer if being used, and further mixing. A ing glycemic control, erectile dysfunction, metabolic gelling agent, if being used, can be introduced under stirring. syndrome, frailty, angina pectoris, congestive cardiac A neutralizer, if being used, usually is added at or near the end failure, osteopenia and osteoporosis or treating erec of the method, such as to the otherwise final composition. For tile dysfunction; and example, if the composition comprises Carbopol R, NaOH or 0176 other therapies involving the systemic admin triethanolamine can be used to neutralize the composition. istration of an androgen. Other optional components can be added at other stages of the 0177. While the foregoing examples have been provided, method, in accordance with known procedures. For example, the skilled artisan readily will appreciate that the composi a preservative, if being used, can be added in an appropriate tions described herein are useful in any context where sys Solvent, at any suitable time of the process. temic delivery of a pharmaceutically active agent, such as one 0185. For example, in a particular embodiment, the com or more Steroids, is desired. Moreover, for any and all uses, ponents may be added and mixed in the following order: one skilled in the art will be able to determine appropriate 0186 1. Add alcohol and co-solvent and mix until uni amounts of gel to apply daily to achieve a target in vivo form. delivery level using a given gel with a given active agent 0187 2. Slowly add therapeutically active agent and mix concentration Such as by using permeation data Such as that until completely dissolved. presented in FIG. 2. 0188 3. Add fatty acid and mix until uniform. 0.178 Exemplary Modes of Administration 0189 4. Add fatty acid ester and mix until uniform. 0179. As noted above, the compositions described herein 0.190 5. Slowly add gelling agent, if being used, and mix are suitable for transdermal administration. For example, the well until completely hydrated. compositions can be directly applied to a Surface of the skin, 0191 6. Slowly add buffer solution, if being used, and mix for direct non-occlusive transdermal/transcutaneous applica until uniform. tion. As used herein, the terms “direct”/"directly” and “non 0.192 The following specific examples are included as occlusive' reflect that the compositions do not require a illustrative of the compositions described herein. These matrix or membrane to effect administration, and thus are not example are in no way intended to limit the scope of the required to be dispensed via a patch, plaster, tape system, or invention. Other aspects of the invention will be apparent to the like. However, the compositions optionally can be dis those skilled in the art to which the invention pertains. pensed via a patch, plaster, tape system or the like. EXAMPLES 0180. The compositions may be administered by any Example 1 means effective to apply the composition to a Surface of the skin. For example, the compositions may be applied manu In Vitro Absorption of Estradiol into the Dermis ally, directly using the hand or with an applicator Such as a 0193 A. Chemicals and Formulations dropper or pipette, an applicator Such as a Swab, brush, cloth, (0194 Tritiated estradiol His used in the preparation of pad, sponge, or with any other applicator, Such as a solid pharmaceutical compositions as below. Support comprising paper, cardboard or a laminate material, including material comprising flocked, glued or otherwise fixed fibers. Alternatively, the compositions may be applied as an aerosol or non-aerosol spray, from a pressurized or Formulation 1 2 3 4 non-pressurized container. In some embodiments, the com Estradiol (E2) (g) O.12 O.12 O.24 O.24 Oleic acid (OA) (g) 2 2 2 2 positions are administered in metered doses, such as from a ethyl oleate (EO) (g) 2 2 metered dose applicator or from an applicator comprising a propylene glycol (PG) (g) 5 5 5 5 single dose of the composition. Ethanol 96% (g) 64 72 64 72 0181. In some embodiments, the composition is adminis Carbonate buffer (CB) (g) 100 100 1OO 1OO tered to a surface of the skin over a defined surface area. The 60 mM, pH 10.7 Qsf administration of a defined, finite amount of the composition to a defined surface area permits the control of the amount of 0.195 The alcohol content is adapted to solubilise the lipo active Substance that is applied to a given Surface area, i.e., philic ingredients. controlling the local concentration. By controlling (e.g., lim (0196. B. Methods 1. Principle of the Method iting) local concentration, local side effects, such as local 0.197 Percutaneous absorption in vitro is studied quanti androgenic effects (including, but not limited to: acne, oily tatively with human skin biopsies placed in Franz diffusion skin), can be minimized. cells (Franz T.J., “Percutaneous absorption on the relevance of 0182. In some embodiments, the amount of composition in vitro data”, J Invest Dermatol. 1975 March; 64(3):190-5) administered is a defined, finite amount that provides a thera permitting contact of a receptor fluid with the dermis in which peutically effective amount (e.g., a single dose) of active the absorbed substance is measured. agent. (0198 2. Description of the Cells 0183 Methods of Making the Compositions 0199 A skin biopsy is maintained horizontally between 0184 Also provided herein are methods for making the two parts of the Franz cell, delimiting two separate compart pharmaceutical compositions. Those skilled in the art can ments referred to as epidermal and dermal. The epidermal US 2011/0098258 A1 Apr. 28, 2011 11 compartment consists of a glass cell cap of precise Surface 0210. 6. Expression of Results Obtained for the Dermis: 0211. The quantity of estradiol which is found in the der area (1.77 cm2), placed on the upper side of the skin. The mis is expressed in ng-equivalent-quantities or in percentages dermal compartment, on the lower side of the skin biopsies, of the administered dose. Each result represents the mean comprises a reservoir of fixed volume (-6.5 ml) fitted with a value of (n) experimental determinations and is associated lateral collection port. The two elements are held in place with with its standard deviation. a clamp. 0212 7. Results and Discussion:

Quantity of % of Estradiol Statistical Estradiol (ng) recovered Mann-Whitney recovered into the into the test N Formulations (n) dermis at 24 H dermis at 24 H * 1 E2 O.12% -- OA 296 - PG S9-0 -- 15 704 244 7.08 it 2.48 P value Ethanol 64% + CB Form. 1. Form. 2 = 0.0004 2 E20.12% -- OA2% -- EO2% -- 16 1219, 418 12.63 - 4.33 PG 5% + Ethanol 72% + CB 3 E20.24% - OA 296 - PG 59-6 -- 16 1557-568 7.79 + 2.83 P value Ethanol 64% + CB Form. 3 Form. 4 = 0.003 4 E20.24% - OA2% -- EO 2% -- 17 2S63 847 13.07 - 4.27 PG 5% + Ethanol 72% + CB * performed on the % of Estradiol recovered in the dermis at 24 h data

0200. The dermal compartment is filled with a receptor 0213. These results show that at both estradiol concentra fluid consisting of a solution of Sodium chloride at 9 g/l and tions tested, the addition of ethyl oleate induces a significant bovine serum albumin at 15 g/l. This liquid is totally removed increase (at least 1.5-fold) in the dermis retention of estradiol periodically throughout the assay and replaced by fresh (p<0.01). (Compare results with Formulation 2 vs. 1 and 4 vs. receptor fluid using the lateral collection port. 3). 0201 A double water-circulation jacket, containing water Example 2 at 37°C., surrounds the lower part of the cell in order to mimic physiologic skin temperature. To ensure the homogeneity of In Vitro Absorption of Testosterone into the Dermis the temperature and the content in the receptor fluid, a stirring 0214 A. Chemicals and Formulations rod is placed in the dermal compartment and each cell is 0215 Tritiated testosterone 3H is used in the preparation placed on a magnetic stirrer. of pharmaceutical compositions as below. 0202 The upper part, or epidermal compartment, is open at the exterior end, exposing the Surface of the skin to the ambient air of the laboratory. 0203 3. Preparation of Skin Biopsies Formulation 1 2 0204 The human abdominal skins used for the experi Testosterone (T) (g) O.24 O.24 oleic acid (OA) (g) 2 2 ments are taken from donors following plastic Surgery proce ethyl oleate (EO) (g) 2 dures. Skins are stored at -20°C. The day before the appli propylene glycol (PG) (g) 5 5 cation of the radioactive formulations, following thawing, Ethanol.96% (g) 64 72 Subcutaneous fats are removed (unless it has already been Carbonate buffer (CB) (g) 100 1OO done before freezing), and the skins are dermatomed at 60 mM, pH 10.7 Qsf approximately 350 Lum. The skins are mounted on the cells the day before application of the radioactive formulation. 0216. The alcohol content is adapted to solubilise the lipo 0205. 4. Operating Procedures philic ingredients. 0206 Ten microliters (s1 uCi) of the preparations are 0217 B. Methods and Results applied over the surface of the epidermis delimited by the 0218. The operating procedures disclosed in Example 1 is glass cell cap. During the experiment, the receptor fluid is followed with the two testosterone formulations described completely removed at 2, 4, 6, 8 and 24 hours through the above. lateral collection port. The dermal compartment is then 0219. After 24 hours, the residual drug remaining on the refilled with fresh solution. surface of the skin is removed by washing cell by cell the surface of the skin just before refilling the dermal compart 0207. At the end of the test (24 hours), the residual drug ment with fresh receptor fluid. The cells are then monitored remaining at the Surface of the skin is removed by washing the for another 24 h. Surface. The epidermis is separated from the dermis by gently 0220. After 48 hours, the receptor fluid is collected and the scraping with a scalpel. epidermis is separated from the dermis by gently scraping 0208 5. Treatment of the Samples and Measurement of with a scalpel. The dermis is separated from the lower part of the Radioactivity the cell. The epidermis and dermis layers are digested for a 0209. The radioactivity contained in the samples obtained few hours at 60° C. for extraction of radioactivity in 1 ml as previously described is measured using a Scintillating liq (epidermis) or in 3 ml (dermis) of Soluene 350TM (PACK uid beta counter equipped with dedicated software. ARD). US 2011/0098258 A1 Apr. 28, 2011

0221) a) Retention in Dermis at 48 h:

Quantity of % of Testosterone Statistical Testosterone (ng) recovered Mann-Whitney recovered into the into the test N Formulations (n) dermis at 48 H dermis at 48 H : 1 T 0.24% - OA2% -- PG 5% + 8 1308 627 6.543.13 P value Ethanol 64% + CB Form. 1. Form. 2 = 0.046 2 T 0.24% - OA2% -- EO 2% -- 8 2O69772 10.543.93 PG 5% + Ethanol 72% + CB * performed on the % of Testosterone recovered in the dermis at 48 h data

0222. These results show that the addition of ethyl oleate induces a significant increase (at least 1.5-fold) in dermal retention (p<0.05), even when measured 2 days after appli cation and 1 day after skin washing. 0223 b)Release into the Reservoir Between 24 hand 48h:

Quantity of Statistical Testosterone (ng) % of Testosterone Mann-Whitney absorbed between absorbed between test N Formulations (n) 24 h and 48 h. 24h and 48 h : 1 T 0.24% - OA2% -- PG 5% + 8 916 - 133 4.57 0.67 P value Ethanol 64% + CB Form. 1. Form. 2 <0.001 2 T 0.24% - OA2% -- EO 2% -- 8 1582 - 292 8.06 - 149 PG 5% + Ethanol 72% + CB

0224. The results also show a significant impact of the ethyl oleate on the penetration absorption 24 hours after skin wash. Formulation 1 2 0225. These results clearly show that the compositions Testosterone (T) (g) O.24 O.24 and methods described herein provide a sustained release of myristic acid (MA) (g) 2 2 testosterone active agent from the skin throughout the 24 isopropyl myristate (IPM) (g) 2 hours after skin wash. propylene glycol (PG) (g) 5 5 isopropanol (g) 56 56 Carbonate buffer (CB) (g) 1OO 100 Example 3 60 mM, pH 10.7 Qsf In Vitro Absorption of Testosterone into the Dermis 0228. The alcohol content is adapted to solubilise the lipo philic ingredients. 0229 B. Methods and Results 0226 A. Chemicals and Formulations 0230. The operating procedures disclosed in Example 1 is 0227 Tritiated testosterone 3H, is used in the prepara followed with the two testosterone formulations described tion of pharmaceutical compositions as below. above.

Quantity of % of Testosterone Statistical Testosterone (ng) recovered Mann-Whitney recovered into the into the test N Formulations (n) dermis at 24 H dermis at 24 H *

1 TO.24% - MA2% -- PG 5% - 9 334 - 172 159 (0.82 P value isopropanol 56% + CB Form. 1. Form. 2 = 0.001 2 T 0.24% - MA2% -- IPM2% -- 8 885 287 4.31 - 140 PG 5% + isopropanol 56% + CB

* performed on the % of Testosterone recovered in the dermis at 24h data US 2011/0098258 A1 Apr. 28, 2011

0231. These results show that at the testosterone concen absorption kinetics (Franz T.J., “The finite dose technique as tration tested, the addition of isopropyl myristate induces a a valid in vitro model for the study of percutaneous absorption significant increase (at least 2.5-fold) in the dermal retention in man.” In: Skin: Drug Application and Evaluation of Envi after 24 hours (p<0.01). ronmental Hazards, Current Problems in Dermatology, vol.7. G. Simon, Z. Paster, M. Klingberg, M. Kaye (Eds), Basel, Example 4 Switzerland, S. Karger, 1978, pp 58-68). In Vitro Absorption of Dihydrotestosterone into the 0239 C. Study Design Dermis 0240. The percutaneous absorption pharmacokinetics of progesterone from two test and one reference formulations 0232 A. Chemicals and Formulations was studied using the invitro finite dose model on human skin 0233 Tritiated dihydrotestosterone 3H, is used in the using a single center, open label, within donor, study of three preparation of pharmaceutical compositions as below. (3) topical gel formulations containing progesterone. Each formulation was tested in triplicate on three different skin donors using the in vitro Franz finite dose skin model. Formulation 1 2 0241. D. Study Products and Dosing 0242 Reference Product: Commercial Progestogel (1% dihydrotestosterone (DHT) (g) 0.7 0.7 progesterone hydroalcholic gel) (Besins Healthcare). Royfirst (IPM) . 7. 5 7. 0243 Test Product(s): Carbopol980 TM (g) 0.5 O.S 0244 New Formulation #1: triethanolamine (TEA) (g) O.S O.S water Qsf (g) 100 1OO

P 190 0234 B. Methods and Results ERS 190 Proof) 729% 0235. The operating procedures disclosed in Example 1 is Propylene Glycol 59% followed with the two DHT formulations described above.

Quantity of % of DHT Statistical DHT (ng) recovered Mann-Whitney recovered into the into the test N Formulations (n) dermis at 24 H dermis at 24 H * 1 DHT gel with 0.5% IPM 7 676- 186 2.50 - 0.69 P value 2 DHT gel With 1.0% IPM 6 1758 - S09 6.67 + .1.93 Form. 1. Form. 2 <0.05 * performed on the % of DHT recovered in the dermis at 24 h data

0236. These results show that an increase of the percent age of IPM in the gel induces a significant (p<0.05) dermal -continued retention of Dihydrotestosterone (at least 2-fold) after 24 Oleic acid 2% hours. Ethyl oleate 2% Pemulen TR-1 2% Example 5 Carbonate buffer (pH 10.8) qsf 100% Evaluation of the Percutaneous Absorption of Progesterone using Franz Human Skin Finite Dose 0245 New Formulation #2: Model 0237 A. Introduction 0238. The in vitro Franz human skin finite dose model has Progesterone 3% proven to be a valuable tool for the study of percutaneous Ethanol (USP 190 Proof) 729% Propylene Glycol 59% absorption and the determination of the pharmacokinetics of Oleic acid 2% topically applied drugs. The model uses human ex vivo Ethyl oleate 2% cadaver or Surgical skin mounted in specially designed diffu Pemulen TR-1 2% sion chambers allowing the skin to be maintained at a tem Carbonate buffer (pH 10.8) qsf 100% perature and humidity that match typical in Vivo conditions (Franz, T.J., “Percutaneous absorption: on the relevance of in vitro data”, J Invest Derm 1975, 64:190-195.). A finite dose 0246 (Carbonate buffer was prepared from 16.91 parts (for example, 4-7 mg/cm2) of formulation is applied to the water, 0.070 parts sodium carbonate and 0.007 parts sodium outer Surface of the skin and drug absorption is measured by bicarbonate.) monitoring its rate of appearance in the reservoir Solution 0247 Dosing bathing the inner Surface of the skin. Data defining total 0248 5 uL formulation/cm2/skin section (dosed by absorption, rate of absorption, as well as skin content can be pipette and rubbed in using a glass rod). The glass rod is accurately determined in this model. The method has historic retained for analysis as part of the mass balance accountabil precedent for accurately predicting in vivo percutaneous ity and for correction of the applied dose. US 2011/0098258 A1 Apr. 28, 2011

0249 E. Study Procedures face of the skin. Following the wash, the intact skin is then 0250) 1. Reagents and Source of Standards removed from the chamber and extracted in 50:50 Methanol: Water. 0251 All reagents used in this study are of analytical 0263 5. Analytical Laboratory reagent grade or better. 0264. Quantification of Progesterone is performed by 0252) 2. Reservoir Medium High Performance Liquid Chromatography (HPLC). Briefly, 0253 For the skin integrity test, the medium base consists HPLC is conducted on a Hewlett-Packard 1100 Series HPLC of phosphate buffered saline (pH 7.4+0.1). For all further system with a diode array UV detector and, if needed, a Mass study conduct, the medium base consists of 0.1x PBS with Spectroscopy (MS) using the current laboratory method. 0.1% Volpo (a non-ionic surfactant: Volpo (Oleth-20) is a Peak areas are quantified to concentration using an external non-ionic Surfactant known to increase the aqueous solubility standard curve prepared daily from the neat standard. of poorly water soluble compounds. Volpo in the reservoir Samples not assayed on the day of collection are stored at or Solution will ensure diffusion sink conditions during percu below -20° C. taneous absorption, and is known not to affect the barrier 0265 F. Analyses and Reports properties of the test skin). 0266 1. Study Parameters 0254 3. Diffusion Cell and Skin Preparation 0267. The following parameters are calculated: 0255 Human, ex vivo trunk skin without obvious signs of 0268 a) Total absorption (sum of all reservoir solutions skin disease is used in this study. It has been dermatomed, sampled from a chamber) cryopreserved, sealed in a water-impermeable plastic bag, 0269 b) Rate and Extent of penetration across the study and stored at ~-70° C. until the day of the experiment. Prior period. to use it is thawed in -37°C. water, then rinsed in tap water to 0270 c) Surface wash and skin content. remove any adherent blood or other material from the surface. 0271 2. Data Evaluation 0256 Skin from a single donor is cut into multiple smaller 0272 a) If any sample is

TABLE 6-3

Total Absorption Across Skin Donors Percutaneous Absorption of Estradiol through Intact Human Cadaver Skin over 48 hours from a Single Application. Mean i SE as Percent of Applied Dose and Total Mass (ug/cm).

Estrogel (R) Estrogel (R) (Lotti (Commercial Parameter Formulation A Formulation B Formulation C Formulation D 769-0929A02) product)

24hr O.835 - O.OO7 O.725 O404 O.993 - 0.608 0.521 - 0.181 (0.031 O.O26 Cumulative Penetration (g/cm) 48 hr 1556 0.239 1570 - 0.674 1765 - 0.862 1031 O.421 O.O71 O.O11 O.065 O.OO8 Cumulative Penetration (Lig/cm) 48 hr 8.643 1.328 8.721 3.746 9.805 - 4.787 5.728 - 2.340 2.358 0.369 2.166 0.255 Cumulative Penetration (%) US 2011/0098258 A1 Apr. 28, 2011

0286 These results show that Formulation C gave maxi mal delivery, about 25-fold greater than the Estrogel(R) for mulation. Formulation C was subsequently studied at lower Estradiol Oleic Propylene Carb. estradiol concentrations to determine a dose-response. The Formulation # Run % Acid 9% Glycol % Buffer % results (FIG. 2) show that the amount of drug delivered 2O6 6 O.OS O.OO 7.OO 6.95 increased with increasing concentration of estradiol applied. 213 3 O.OO 7.OO 6.95 The results obtained at the highest concentration (0.36%) 214 4 3.47 3.53 6.95 agree well with the data obtained in this example (Table 6-3, 208 8 7.OO O.OO 6.95 216 6 7.OO O.OO 6.95 Formulation C). In addition, the values obtained for Estro 211 1 O16 140 5.60 6.84 gel(R) (0.06% estradiol) are in close agreement between the 215 5 4.90 2.10 6.84 two studies, further supporting their reliability. 205 5 O.26 7.OO O.OO 6.74 210 O 7.OO O.OO 6.74 0287. The results depicted in FIG. 2 also show that even at 2O7 7 O.28 O.OO 7.OO 6.72 roughly equivalent estradiol concentrations (0.07% vs. 2O1 1 3.35 3.65 6.72 0.06%), the new formulation (C) delivered about 10-fold 212 2 3.SO 3.SO 6.73 219 9 O.38 5.60 140 6.62 more drug than the Estrogel(R) formulation. Thus, the compo 204 4 O41 1.46 S.S4 6.59 sitions described herein make it possible to deliver an equiva 218 8 3.SO 3.SO 6.59 lent dose to the existing commercial transdermal gel product 2O3 3 O.SO O.OO 7.OO 6.50 220 2O O.OO 7.OO 6.50 with 10 times less applied volume, such as with Formulation 217 7 3.21 3.79 6.50 C containing estradiol at 0.07%. This represents a significant 2O2 2 7.OO O.OO 6.50 advantage, including safety advantages, regulatory advan 209 9 7.OO O.OO 6.50 tages and cost savings due to the need for so much less product to provide an equivalent dose. For example, regula 0296. Additionally, the following steps are performed: tory agencies often encourage the development of products 0297 Potency Assessment that contain a minimal amount of active agent required for 0298. The Estradiol potency of the final formulations can therapeutic efficacy. be determined in triplicate by HPLC/UV. Potency can be calculated as (w/v) to calculate the mass amount of Example 7 0299 Estradiol in the applied dose, so that the percentage absorbed of the applied dose can be calculated. Potency also Evaluation of Formulation Influences on Percutane can be calculated, correcting for density, as (w/w), to compare ous Absorption of Estradiol using Franz Human Skin with the target potency of the prepared formulations indicated in the table above. Estradiol Potency must be within +5.0% to Finite Dose Model be acceptable for this study. In the data analysis the actual estradiol concentrations of each formulations are used. 0288. In order to study the influence of the penetration (0300 Formulation Preparation: enhancers and of the co-solvent on the percutaneous absorp 0301 1. Add ethanol and propylene glycol and mix until tion of the active in the new gel formulations, a 2-phase, uniform. statistically designed, experiment was conducted. 0302) 2. Slowly add estradiol and mix until completely 0289. In the first phase, the influence of varying oleic acid dissolved. and co-solvent (propylene glycol) concentrations, together 0303 3. Add oleic acid and mix until uniform. with the estradiol concentration, on the total amount of active 0304 4. Add ethyl oleate and mix until uniform. delivered was studied. (0305 5. Slowly add Pemulen TR-1 and mix well until 0290. In the second phase, the influence of varying the completely hydrated. ethyl oleate and the estradiol concentrations on the amount of (0306 6. Slowly add the carbonate buffer solution to the active delivered and on the time profile of the delivery was above gel matrix and mix until uniform. studied. (0307 B. Second Phase of the Study 0291. The “Design-Expert” statistical software (available 0308 For the second phase, a Response-surface design from StatEase at www.statease.com) was used to generate the with a Central Composite Structure is used. experimental data points used in the study. 0309 The formulations to be studied are shown in the 0292. The same protocol as that described in Example 5 is following table, with the addition of ethanol at 72%, followed, with differences noted below. Pemulen TR-1 at 2%, oleic acid at 2%, and propylene glycol 0293 A. First Phase of the Study at 5%. 0294 For the first phase, a combined D-optimal design with oleic acid and propylene glycol as mixture components and with estradiol as numeric factor (process) is used. The Formulation A: Estradiol B: EthylOleate Carb. Buffer oleic acid and propylene glycol concentrations are varied in i Run % % % Such away that the total of their two concentrations remained 305 5 O.OS 1.00 17.95 constant and equal to 7%, thus minimizing potential differ 311 11 O.12 O.29 18.59 ences in solubility between formulations. 3O2 2 1.71 17.18 303 3 O.28 O.OO 18.73 0295 The Table below summarizes the formulations that 304 4 1.00 17.73 are prepared and tested in triplicate on samples from two 307 7 17.73 different donors. Each formulation contained in addition: 3O8 8 17.73 ethanol at 72%, ethyl oleate at 2% and Pemulen TR-1 at 2%. US 2011/0098258 A1 Apr. 28, 2011 17

experimental data points is not good, however, as can be seen -continued in FIG.3 for the replicates conducted at 0.26 and 0.5% estra diol. This lack of reproducibility is confirmed by looking at Formulation A: Estradiol B: Ethyl Oleate Carb. Buffer i Run % % % the standard error graph for the same dataset (not shown— error increases with increasing oleic acid concentrations), 310 10 2.OO 16.73 and also at the individual data for each experimental point, 301 1 O.43 O.29 18.27 306 6 1.71 16.86 which consisted of 3 replicates on two different donors. Such 309 9 O.SO 1.OO 17.50 a spread in absorption from one donor sample to the other, and even between replicates on the same donor sample indicates instability in the system. Indeed, some experiments deliver 0310. The rest of the experimental procedure is as that of large amounts of active while other experiments, despite all the first phase of the study. experimental parameters being kept constant, deliver signifi 0311. C. Results from the First Phase of the Study cantly less active. Such behaviour is not desirable in a phar 0312. As described in Section A. above under the para maceutical composition, because when translated into the graph “Potency Assessment each formulation is checked for clinic, these formulations could give large patient-to-patient its actual estradiol concentration. The measured values are variations or even within-patient variations from application shown in the Table below, and are used in the data analysis. to-application. For these reasons, it may be advantageous to select ranges offatty acid and co-solvent concentrations that do not encompass the highest and lowest propylene glycol/ F # Target Conc (%) Measured Conc (%) oleic acid concentrations studied here. 0318. Further illustrating this point, FIG. 4 represents a 2O1 O.28 O.29 2O2 OSO O48 top-down view of the data illustrated three-dimensionally in 2O3 OSO O48 FIG.3. The middle region, centered around 2% oleic acid and 204 O41 O43 5% propylene glycol, appears to be the most desirable for a 205 O.26 O.24 composition that exhibits absorption with minimal depen 2O6 O.OS O.OS 2O7 O.28 0.27 dency on active agent concentration (the problem seen with 208 O.OS O.OS higher concentrations of propylene glycol) while achieving 209 OSO O49 strong reproducibility between data points (contrary to the 210 O.26 O.24 211 O16 O16 data obtained with higher concentrations of oleic acid), even 212 O.28 O.28 though not providing the highest delivery of active. 213 O.OS O.OS 0319. In specific embodiments, therefore, the fatty acid 214 O.OS O.O6 permeation enhancer is present in an amount of from 0.01% 215 O16 O16 216 O.OS O.04 to 5%, including from 0.05% to 3.5%, such as 1% to 3%, by 217 OSO OSO weight based on the total weight of the composition. 218 O41 O42 0320 Additionally, in specific embodiments, the co-sol 219 O.38 0.37 vent (Such as propylene glycol) is present in an amount of 220 OSO O42 from 0.01% to 7%, including from 3% to 7%, such as from 4% to 6%, by weight based on the total weight of the com 0313 The total penetration data (amount of active having position. penetrated in the reservoir compartment after 48 hours) was of sufficient quality to allow analysis in a statistically signifi 0321) D. Results from the Second Phase of the Study cant manner with quadratic models for the mixture and pro 0322 FIGS. 5 and 6 illustrate the influence of ethyl oleate cess parts of the design. and estradiol concentration on the total absorption over 48 0314 FIG. 3 illustrates the response surface obtained. As hours. The variation of estradiol concentration affects the propylene glycol (front of the graph) is gradually replaced by absorption in a bell-shaped fashion, as already illustrated in oleic acid (back of the graph), the variation of absorption as a the first phase of the study at the corresponding oleic acid and function of the estradiol concentration goes from a bell propylene glycol concentrations. The addition of ethyl oleate shaped curve to a flat, constant curve. has the effect of increasing the total amount of absorption and 0315. A bell-shaped curve results from a strong depen also of shifting the optimal estradiol concentration (i.e. the dency of the absorption on the concentration of therapeuti estradiol concentration corresponding to maximum absorp cally active agent in the formulation, as is the case, for tion) to higher values. This phenomenon is most clearly vis example, for formulations having high propylene glycol and ible in FIG. 6. low oleic acid concentrations. 0323. The main effect of the ester(ethyl oleate), however, 0316. This dependency on the concentration of therapeu as already described in examples 1 to 4, is to modify the tically active agent is not desirable, since compositions able to delivery profile over time, providing a Sustained release achieve efficient delivery over greater ranges of active agent effect. To illustrate this phenomenon, the graphs representing concentrations are generally preferable from a regulatory and time courses of the absorption fluxes for the 11 compositions commercial perspective. tested have been grouped in 3 categories, illustrated in FIGS. 0317. At the other end of the oleic acid/propylene glycol 7 to 9. axis, i.e. in the high oleic acid/low propylene glycol region, 0324 FIG. 7 shows flux profiles for a first group which the dependency of the absorption on the estradiol concentra trigger after about 20 hours an increase in flux, leading to a tion is not observed, and the total absorption reaches higher profile where the dose escalates with time. This is not desir absolute levels, most likely due to the efficiency of the fatty able for a product where a strong delivery within hours of acid as penetration enhancer. The reproducibility of the application is sought followed by a plateau of steady release US 2011/0098258 A1 Apr. 28, 2011

of the drug. The three data points illustrated in FIG. 7 all for widespread clinical use, skin sensitization studies are belong to the “low ethyl oleate”-"highestradiol corner in the conducted in guinea pigs and rabbits, using the following graph from FIG. 6. formulations: 0325 FIG. 8 displays flux profiles of a second group, 0335 K36 Active: where the flux decreases rapidly after the peak occurring at 6 hours post administration. This profile is typical of a number of prior art compositions, which achieve fast and efficient delivery within the first few hours post administration, but Chemical Name % Wiw which lack steady release overalonger term, i.e. 24 or even 48 Purified water USP 16.91 hours. The three data points illustrated in FIG. 8 are along the Alcohol USP 190 Proof 71.95 Propylene Glycol USP/EP S.OO Y=X line in the graph from FIG. 6. Super Refined Oleic Acid NF 2.OO 0326 Finally, FIG. 9 displays profiles of experimental Ethyl Oleate NF 2.OO data points with an early, rapid rise in flux, followed by a Estradiol USP O.36 steady flux level over 2 days, i.e., a Sustained release, storage Hydroxypropyl cellulose NF (Klucel HF) 1.70 depot effect. This type of flux is desirable in many therapies, Sodium Bicarbonate USP O.OO7 where both rapid attainment of therapeutic blood concentra Sodium Carbonate NF O.O7 tions and Sustained blood concentrations of drug are desired. The compositions that achieve this type of profile are com 0336 P36 Active: positions in the “high ethyl oleate”-"low estradiol half of the graph from FIG. 6 (in other words, above the Y=X line). 0327. This qualitative analysis of the flux profiles over time demonstrates that a Sustained release of active agent is Chemical Name % Wiw achieved in a satisfactory manner when the fatty acid ester is Purified water USP 16.91 present in a greater amount than the active agent, Such as the Alcohol USP 190 Proof 71.65 Propylene Glycol USP/EP S.OO fatty acid ester being present in an amount at least four times Super Refined Oleic Acid NF 2.OO greater than that of the active agent, on a weight: weight basis. Ethyl Oleate NF 2.OO 0328. In particular, it appears that the more ester is present Estradiol USP O.36 in the composition, the better the storage depot effect is. This Pemulen TR-1 2.OO factor Suggests that one should aim to include as much fatty Sodium Bicarbonate USP O.OO7 acid ester in the composition as possible. An upper limit is Sodium Carbonate NF 0.07 imposed, however, by the solubility of the fatty acid ester in the composition. As an example, FIG. 10 illustrates the 0337 K36 Placebo: amount of ethyl oleate that can be dissolved, at room tem perature, as a function of the ethanol (96% V/v) concentration in a formulation made up with: 0329 0.24% estradiol: Chemical Name % Wiw 0330) 5% propylene glycol; and Purified water USP 16.91 Alcohol USP 190 Proof 72.30 0331, 2% oleic acid Propylene Glycol USP/EP S.OO 0332 Qsf water. Super Refined Oleic Acid NF 2.OO Ethyl Oleate NF 2.OO 0333. From FIG.10 and the Table below, it is apparent that Hydroxypropyl cellulose NF (Klucel HF) 1.70 in a formulation comprising 72% ethanol, a maximum of Sodium Bicarbonate USP O.OO7 2.2% ethyl oleate can be dissolved. Sodium Carbonate NF O.O7

0338 P36 Placebo: EtOH (96% w/v) concentration Amount of Ethyl Oleate in the mixture solubilised in g100 g

64% 0.65 g/100 g Chemical Name % Wiw 66% 0.91 g/100 g 68% 1.28 g/100 g Purified water USP 16.91 70% 1.60 g/100 g Alcohol USP 190 Proof 72.OO 729% 2.19 g/100 g Propylene Glycol USP/EP S.OO 7396 2.40 g/100 g Super Refined Oleic Acid NF 2.OO Ethyl Oleate NF 2.OO Pemulen TR-1 2.OO Sodium Bicarbonate USP O.OO7 Example 8 Sodium Carbonate NF O.O7 Skin Sensitization Studies 0339. These studies are conducted to evaluate the potential 0334 Previous studies have reported skin irritation prob of the test compositions, K36 Active and P36 Active, to cause lems with transdermal compositions comprising high or elicit skin sensitization reactions (allergic contact derma amounts of co-solvents, such as propylene glycol, at the titis) via topical patch applications in animal models. amounts used in some embodiments described herein, Such as 0340 Guinea Pigs: The compositions are applied by at about 5% (w/w). To determine whether the compositions closed topical patch and Hilltop Chamber application to Crl: described herein are irritating, and thus possibly not suitable HA (Albino Hartley) Guinea Pigs. During the induction US 2011/0098258 A1 Apr. 28, 2011

phase, three treatment groups of five animals/sex/group were observed for all three animals and was considered to be test administered K36 Placebo, P36Placebo, or the positive con article related, but not adverse. trol, Hexylcinnamic aldehyde (HCA 100%), while the 0346. These studies demonstrate that the compositions remaining two treatment groups often animals/sex/group are described herein, comprising about 5% propylene glycol, are administered the test compositions, K36 Active or P36 not irritating, and do not give rise to significant skin sensiti Active. During the challenge phase, each placebo group is zation effects. Thus, these factors would not limit their clini administered the respective test composition, and the positive cal use. control receives 50% HCA in mineral oil (HCA50%). During both phases, all groups are administered the placebos, posi Example 9 tive control, or test compositions by dermal application at 0.4 21-Day Dermal Toxicity Study in Rabbits mL/dose. During the induction phase, the placebos, positive control and test compositions are administered once a week 0347 This study is conducted to evaluate the potential for 3 weeks on Days 1, 8 and 15, followed by a 2 week toxicity of the two formulations of the test compositions washout period, while during the challenge phase, the posi described above, K36 Active and P36 Active, and their respective placebos when administered once a day via dermal tive control and testarticles are administered once on Day 29. application for 21 consecutive days to two treatment groups 0341 For the duration of the study, observations for mor often male and ten female New Zealand White Hra:(NZW) bidity, mortality, injury, and the availability of food and water SPF albino rabbits. are conducted twice daily for all animals. In addition, body 0348 Both the K36 Active and the P36 Active are formu weights for all animals are measured and recorded prior to lated at an active concentration of 0.36% estradiol. The test randomization (Day -7), prior to each test compositions articles are administered at a dose Volume of approximately administration (with the exception of Day 15, body weights 0.85 to 1.11 mL. Two additional groups often animals/sex were recorded approximately 6 hours postdose after unwrap will serve as the control and receive the placebos, K36 Pla ping), and the day prior to termination (Day 31). During the cebo and P36 Placebo. challenge phase only, dermal irritation scoring for skin sen 0349. Observations for morbidity, mortality, injury, and sitization is conducted at approximately 24 and 48 hours after the availability of food and water are conducted twice daily patch removal (post dose). At study termination, the animals for all animals. Clinical observations are conducted weekly. are euthanized by carbon dioxide inhalation. Test sites are evaluated for erythema and edema daily during 0342. Dermal irritation scores recorded at 24 and 48 hours the first week of dosing, and weekly thereafter. Body weights post dose during the challenge phase indicated that sensitiza are measured and recorded weekly. Food consumption is tion did not occur following the administration of induction measured and recorded daily. Blood and urine samples for doses and Subsequent two week washout period. Irritation clinical pathology evaluations are collected from all animals scores in the K36 Active and P36 Active groups were gener pretest and prior to the terminal necropsy. At Study termina ally equivalent or lower than scores recorded for the K36 tion, necropsy examinations are performed and organ weights Placebo and P36Placebo groups. Additionally, reduced body are recorded. Selected tissues are microscopically examined weight gain was observed in the K36 Active and P36 Active for animals that received P36 Active and Placebo. Tissues when compared with the respective placebo groups. These from the other two groups on study are held for possible lower body weight gains were considered to be test article future reference. related but not adverse. 0350. There was no test composition-related change in 0343 Rabbits: This study is conducted to evaluate the body weight and no clear test composition-related clinical potential dermal irritant and/or corrosive effects of the test findings. Possible test composition-related clinical findings compositions. One treatment group of three female New included inappetance, aggressive behavior, and Vocalization. Zealand White Hra:(NZW)SPF albino rabbits is administered These findings were limited to one animal per sex per group active formulations K36 and P36, and their respective place treated with either K36 Active or P36 Active, and were not bos, to one of the four dorsal sites at a dose level of 0.5 observed in either placebo group. mL/site. The placebos and testarticles are administered to the 0351 Very slight (barely perceptible) erythema was respective test sites on each animal via dermal application, observed sporadically throughout the study with similar or once daily, for 3 consecutive days. lower frequency in the K36 Active and P36 Active than in the 0344 Observations for morbidity, mortality, injury, and respective placebo groups. These findings were considered to the availability of food and water are conducted twice daily be primarily related to the vehicle and not test composition for all animals. Body weights are measured and recorded related. predose. Dermal irritation scores are conducted within 30-60 0352 Test composition-related, but not adverse, decreases minutes, and at 4 and 24 hours post dose on Days 1 and 2. On in food consumption were observed in both the K36 Active Day 3, the test sites are scored within 30-60 minutes, and at 4, and P36 Active groups when compared with the respective 24, 48, and 72 hours post dose. Additional irritation scores are placebo groups. Male food consumption was more severely conducted on Days 8 and 15 to fully evaluate the reversibility affected (18.4%-19.2%) and frequently statistically signifi or irreversibility of the effects observed. At study termination, cant, whereas female food consumption was moderately all animals are euthanized, and the carcasses are discarded affected (6.5%-11.1%) and only occasionally statistically without further evaluation. significant. 0345 Minimal to mild erythema and edema was observed 0353 Test composition-related changes in hematology for both K36 and P36 Placebo and Active formulations. P36 parameters included moderate decreases in erythrocytes, Placebo and P36 Active appeared to cause slightly more irri hemoglobin, hematocrit, reticulocytes, and platelets in both tation than K36 Placebo and K36 Active, though these differ the K36 Active and P36 Active dose groups. Total leukocytes ences were minimal. A slight decrease in body weight was and lymphocytes were also decreased in these groups. US 2011/0098258 A1 Apr. 28, 2011 20

0354 Test composition-related changes in clinical chem 0362 Microscopic alterations within the prostate gland istry parameters included increased aspartate aminotrans and seminal vesicles of treated animals included hypertrophy ferase (AST), alanine aminotransferase (ALT), Y-glutamyl of the Smooth muscle associated with both of these glands, transferase (GGT), sorbitol dehydrogenase (SDH), urea and occasional animals had increased fibroplasia with the nitrogen, and creatinine, and decreased triglycerides in both subepithelial stroma resulting in thickening of the intraglan the K36 Active and P36 Active groups. The increases in the dular septa in the prostate gland. In addition to these stromal liver enzymes tended to be slightly greater in the males changes, there was a dysmaturity or regression of the glan receiving P36 than in those receiving K36. The increases in dular epithelium, meaning that the epithelium was of urea nitrogen and creatinine were minimal and may have been secondary. decreased maturity (increased immaturity) compared to other 0355 There were no test composition-related changes in features of the gland Such as increased luminal diameter. coagulation or urinalysis parameters. 0363 Within treated females, there was mild to severe 0356. There were no test composition-related macro mucification of the vaginal epithelium and mild to moderate scopic findings in the males in this study. Test composition hypertrophy of the vaginal Smooth muscle. A similar finding related macroscopic observations in females in the P36 was present in the epithelium of the cervical region of the Active group included oviduct cysts in three animals and an uterus. A decidual reaction (Zook, B.C., O. A. Janne, A. A. abdominal cavity adhesion in which the uterus and cervix was Abraham, and H. A. Nash. “The Development and Regres adhered to the abdominal wall in one animal. A likely test sion of Deciduosarcomas and Other Lesions Caused by composition-related macroscopic observation of red discol Estrogens and Progestins in Rabbits. Toxicologic Pathology oration of the uterine horn and body was made in one female 29.4 (2001): 411-416: Jaane, O.A., B. C. Zook, A. K. Didol rabbit in the K36 Active group. Though microscopic analysis kar, K. Sundaram, and H. A. Nash. “The Roles of Estrogen was not performed, it is likely that this observation correlated and Progestin in Producing Deciduosarcoma and Other to lesions similar to those seen in the P36 Active female Lesions in the Rabbit.” Toxicologic Pathology 29.4 (2001): rabbits. 417-421), a common response to estrogenic compounds, was 0357 Test composition-related statistically significant seen to at least a minimal degree in all uterus samples and two alterations in organ weights occurred in the liver, spleen and spleens from the P36 Active group females. The decidual thymus weights of both males and females in both the P36 reaction occurs both within the subendometrial stroma as Active and K36 Active groups, and in the uterus with cervix well as the blood vessels within the uterus. In rabbits with weights of the females from both the P36 Active and K36 severe affected vessels, there was associated ischemic necro Active groups. sis of the adjacent uterine tissue due to the disrupted blood 0358 Test composition-related microscopic alterations Supply. This necrosis was occasionally transmural, and in one occurred within the liver, spleen and thymus of males and rabbit, resulted in fibrosis on the abdominal surface of the females, the prostate and seminal vesicles in males, and the uterus and adhesion to the parietal surface of the abdominal Oviducts, uterus with cervix and vagina of females. cavity. 0359. Within the liver, there was a diffuse depletion of intrahepatocellular glycogen stores. In addition there was a 0364. Also in females, three of the animals in the P36 minimal to mildbile duct hyperplasia. The biliary hyperplasia Active group had cysts on the Oviducts. Cysts are not uncom was possibly a direct testarticle effect, as estrogens have been mon in various female reproductive organs, so it is possible shown to stimulate cholangiocyte proliferation (LeSage, G., that these cysts may represent developmental anomalies, S. Glaser and G. Alpini. “Regulation of Cholangiocyte Pro however the presence of these cysts in three P36 Active group liferation.” Liver 21 (2001): 73-80.). females and no control animals suggests that this alteration 0360. Within the spleen, there was a minimal to moderate may be related to the administration of the test article. hyperplasia of the reticuloendothelial macrophages which 0365. There were no significant differences between the was occasionally accompanied by increased erythrophagocy K36 Active and P36 Active treatment groups. tosis, an increase in pigmented (hemosiderin-laden) mac 0366. In general, these findings are commonly associated rophages and rarely by dilation of the splenic red pulp sinu with administration of test composition containing estrogens, soids. In addition, there was minimal to moderate depletion of the splenic lymphoid population in treated animals. These and so do not undermine the potential clinical usefulness of alterations may both be direct test composition effects as the specific formulations described herein. estrogens have been shown in rats to stimulate reticuloendot helial cells of the spleen resulting in increased phagocytosis Example 10 (Steven, W. M. and T. Snook. “The Stimulatory Effects of Diethylstilbesterol and Diethylstilbesterol Diphosphate on Dose-Escalation Study in 12 Healthy Human Male the Reticuloendothelial Cells of the Rat Spleen.” American Subjects Journal of Anatomy 144.3 (1975): 339-359), and also, high levels of estrogens have been shown to cause decreases in 0367. A multiple-dose, open-label, dose-escalation study both T- and B-cell populations within the spleen of rats was designed in which 12 healthy male Subjects were sched (Burns-Naas, L. A. B. J. Meade and A. E. Munson. “Toxic uled to receive 1 of 2 treatments once daily for 3 days with an Response of the Immune System.” Cassarett & Doull's Toxi 11-day washout period between doses. cology. The Basic Science of Poisons. Ed. Curtis D. Klaassen. 0368. The objective of this Phase 1 study was to assess the New York: McGraw-Hill, 2001. 419-470.). safety and pharmacokinetic (PK) profile of multiple dose 0361 Thymic changes consisted of mild to severe gener administration of 0.25 g and 1.00 g of a 0.07% transdermal alized lymphoid depletion. Estrogens have been shown to estradiol gel in healthy male Volunteers. The transdermal gel cause thymic depletion (Burns-Naas, Supra). used had the following formulation: US 2011/0098258 A1 Apr. 28, 2011 21

0375. In general, estradiol Tmax after 1.00 g was shorter than that after 0.25 g. Estradiol Tmax was observed between approximately 3 hr (0.25 g, third dosing interval) and 13 hr O.07% estradiol 2.0% ethyl oleate (0.25 g, first dosing interval) after administration of the gel. 2.0% oleic acid 0376. Within each dose group, mean estimates of AUC for S.O% propylene glycol each 24-hr dosing interval (AUC0-24) were comparable, 16.91% purified water 71.94% ethanol indicating no significant accumulation for estradiol. There 2.0% pemulen TR-1 Carbomer was a less than proportional increase in exposure to estradiol O.07% Sodium carbonate anhydrous with an increase in the gel dose. O.OO7% Sodium bicarbonate anhydrous 0377 The increase in peak and overall systemic exposure to estradiol after a 4.00-fold increase in gel dose was 1.95 0369 Twelve (12) healthy subjects were enrolled and par fold based on Cmax0-120 and 1.26-fold based on AUC0-120. ticipated in two open-label treatment periods. Subjects who 0378. The ratios (0.25 g:1.00 g) (90% confidence inter Successfully completed the screening process checked into vals) for estradiol Cmax0-120 and AUC0-120 were 47.89% the research center on Day 1, approximately 1 to 2 hours prior (40.17%, 57.10%) and 80.01% (70.45%, 90.88%), respec to the first blood draw of each treatment period. For Treatment tively. Period A, 0.25g of the 0.07% gel was to be administered once daily for three days. During Treatment Period B, 1.0 g of the Example 11 0.07% gel was to be administered once daily for three days. Dosing days were to be separated by a washout period of at Dose-Escalation Study in 12 Healthy Human Male least 11 days. Subjects 0370 Diagnosis and Main Criteria for Inclusion: Healthy adult male volunteers, 18–45 years of age, with body mass 0379 The primary objective of this Phase 1 study was to index (BMI) between 18 and 30 kg/m2, inclusive, and mini compare the pharmacokinetic (PK) profiles of two different mum weight of 50 kg (110 pounds). formulations of transdermal estradiol gel in healthy male 0371 Results: volunteers. The secondary objective of this study was to 0372 Synopsis Table 1: Parameters of Estradiol assess the incidence and severity of adverse events.

Dose 0.25 g 1.00 g N Mean SD CV% in Mean SD CV% Tmax0-24 (hr) 2 12.83 10.14 79.04 6.91 S.97 86.33 Tmax24-48 (hr) 2 32.67 9.59 29.34 28.18 2.89 10.24 Tmax48-72 (hr) 2 S1.17 3.64 7.11 S4.91 6.16 11.21 Tmax0-120 (hr) 2 33.58. 18.68 SS.63 29.82 20.66 69.28 Tmax-overall (hr) 2 54.83 S6.78 103.56 29.82 20.66 69.28 Cmax0-24 (pg/mL) 2 26.4 7.35 27.83 56.5 20.8 36.74 Cmax24-48 (pg/mL) 2 29.7 113 38.04 S2.1 27.3 S2.43 Cmax48-72 (pg/mL) 2 28.5 15.1 52.95 49.2 22.5 45.73 Cmax0-120 (pg/mL) 2 33.1 16.5 SO.O2 64.6 27.1 42.04 Cmax-overall (pg/mL) 2 33.1 16.5 49.85 64.6 27.1 42.04 AUCO-24 (hr * pg/mL) 12 476.7 115.2 24.16 752.2 238.9 31.76 AUC24-48 (hr * pg/mL) 12 502.7 135.9 27.04 661.O 249.O 37.67 AUC48-72 (hr * pg/mL) 12 476.8 156.8 32.88 658.1 2S2.S. 38.36 AUCO-120 (hr * pg/mL) 12 2333 642.1 27.52 2933 889.1 30.31 AUCO-t (hr * pg/mL) 12 3778 1260 33.35 4436 1204 27.14 Tlast (hr) 2 200.27 37.50 18.73 216.OO O.OO O.OO Clast (pg/mL) 2 17.3 3.87 22.37 17.2 S.O9 29.53

0373 Conclusion: 0380. This was a multiple-dose, open-label, two-treatment 0374 Mean baseline concentrations of estradiol ranged study that has been conducted in two parts to evaluate the pharmacokinetics of estradiol after transdermal administra from 14.3 pg/mL to 21.7 pg/mL. After administration of 0.25 tion of two different formulations to healthy male volunteers. g of gel, estradiol concentrations increased slightly above 0381 12 healthy adult male subjects participated in 2 ran baseline (highest mean plasma concentration 25.6 pg/mL). domized treatment periods. During each Treatment Period, After administration of 1.00 g of gel, estradiol concentrations either 1.25 g of 0.06% EstroGel(R) (estradiol gel) or 1.0 g of a increased by approximately 2- to 3-fold (highest mean 0.07% estradiolgel according to the invention (formulation in the Table below) were administered once daily to the subject's plasma concentration 54.3 pg/mL). In general, concentra upper arm for five days. Subjects were randomized to receive tions of estradiol returned to baseline levels at approximately either Treatment Code A or B during the first treatment period 12 hr post-dose for the 0.25 g of gel; for the 1.00 g of gel, and the opposite treatment code during the second treatment concentrations of estradiol returned to baseline levels at period. Treatment periods 1 and 2 were separated by a 3 day approximately 96 hr and 120 hr. washout period. US 2011/0098258 A1 Apr. 28, 2011 22

0382. Subjects remained confined in the research center 0386 Results: for blood draws for at least 24 hours after the last study gel 0387. A summary of pharmacokinetic parameters for application in each treatment period, and returned to the estradiol after topical application of the test gel at 0.7 mg research centerfor blood draws and other study procedures on QDx5 days and EstroGel 0.75 mg QDx5 days to healthy male Days 7 and 8. volunteers is provided in the table below.

O.07% estradiol Test Gel EstroGel O.3% ethyl oleate O.3% oleic acid Estradiol (0.7 mg QD x 5 days) (0.75 mg QD x 5 days) 0.75% propylene glycol Cmax (pg/mL) 65.0 + 19.4 (10) 55.6+18.2 (10) 16.91% purified water Tmax (h) 74.0 (10) 85.5 (10) 79.59% ethanol 8.0-98.0 2.0-112 2.0% pemulen TR-1 Carbomer AUC(O - t) 4,131 + 1,194 (10) 4,043 + 654 (10) O.07% Sodium carbonate anhydrous (h x pg/mL). O.OO7% Sodium bicarbonate anhydrous *Arithmetic meant standard deviation (N) except Tmax for which the median (N) Range is reported, Absolute Cmax and Tmax across all doses. AUC from the first through last doses. 0388 Both Examples 10 and 11, where the gels disclosed Treatment Code Treatment Intervention herein were tested in human clinical trials, demonstrated that A: Drug: EstroGel (R) (0.06%) the formulations are safe and effective at delivering the drug Dose = 1.25 gx 0.6 mg g = 0.75 mg estradiol of interest systematically to patients. Topical, once daily for five days 1. A Sustained release pharmaceutical composition for 24 hours between dose applications Application site: upper arm topical administration to a skin Surface comprising: Target plasma level: approximately 80 pg/mL a pharmaceutically active agent comprising one or more steroids, a fatty acid ester, Water, a C2-C6 monoalcohol and a fatty acid, Treatment Code Treatment Intervention wherein the weight: weight ratio of the fatty acid ester in the B: Drug: 0.07% estradiol test gel composition to the total active agent in said composition Dose = 1.0 g x 0.7 mg g = 0.7 mg estradiol Topical, once daily for five days is at least 4:1 fatty acid ester:active agent. 24 hours between dose applications 2. The composition of claim 1, further comprising a co Application site: upper arm solvent. Target plasma level: approximately 80 pg/mL 3. The composition according to claim 1, wherein the co solvent is present in an amount ranging from 0.01% to 7% by weight of the total weight of the pharmaceutical composition. 0383. During each treatment period, blood samples were 4. The composition according to claim 1, wherein the fatty collected for measurement of estradiol prior to each dose and acid esteris selected from the group consisting of ethyl oleate, following each dose at selected times through 72 hours post isopropyl oleate, isopropyl myristate, isopropyl isostearate, dose. Up to 102 blood samples (~510 mL whole blood) were isopropyl palmitate, ethyl octanoate, ethyl dodecanoate, ethyl obtained from each Subject for pharmacokinetic assessments, linoleate, ethyl palmitoleate, ethyl isostearate and ethyllino excluding screening and post-treatment safety assessments. lenate. During Periods 1 and 2, all blood samples were collected 5. The composition according to claim 1, wherein the fatty from the contralateral arm on which the study gel is being acid ester is isopropyl myristate. administered, to prevent sample contamination. 6. The composition according to claim 1, wherein the fatty 0384 Prior to progression to the next treatment period, acid ester is ethyl oleate. safety data (adverse events, clinical laboratory tests) will be 7. The composition according to claim 1, wherein the fatty reviewed before dosing commences. acid ester is the ester that would result from the reaction of the fatty acid formulated in the composition with an alcohol. 0385 For all treatment periods, subjects were required to 8. The composition of according to claim 1, wherein the shower/bathe approximately 1 hour prior to each application fatty acid ester is not the ester that would result from the of the study gel. Showering/bathing privileges were sus reaction of the fatty acid formulated in the composition with pended until 1 hour before the next application of the study the alcohol formulated in the composition. gel. Each dose was applied topically to the designated appli 9. The composition according to claim 1, wherein the fatty cation site. After dosing, no food was allowed until two hours acid ester is present in an amount ranging from 0.01% to 5% post-dose. Water was withheld for one hour post dose and by weight of the total weight of the pharmaceutical compo then allowed ad lib for the remainder of the confinement sition. period. Subjects were served meals at approximately the 10. The composition according to claim 1, wherein the same time relative to dose for each treatment period; the same fatty acid is a C8-C22 fatty acid selected from the group menu choices were available during all treatment periods. consisting of capric acid, lauric acid, myristic acid, palmitic Bathroom/washroom privileges were suspended for one hour acid, Stearic acid, oleic acid, isoStearic acid, palmitoleic acid, after dosing. linoleic acid and linolenic acid. US 2011/0098258 A1 Apr. 28, 2011

11. The composition according to claim 1, wherein the 18. The composition according to claim 1, wherein the fatty acid is present in an amount ranging from 0.01% to 5% alcohol is selected from the group consisting of ethanol, by weight of the total weight of the pharmaceutical compo n-propanol, isopropanol, n-butanol, isobutanol, tert-butanol, sition. and mixtures thereof. 12. The composition according to claim 1, comprising 2% 19. The composition according to claim 1, wherein the ethyl oleate as the fatty acid ester, 2% oleic acid as the fatty acid, and 5% propylene glycolas the co-solvent, all by weight alcohol is present in an amount ranging from 10% to 90% by of the total weight of the pharmaceutical composition. weight of the total weight of the pharmaceutical composition. 13. The composition according to claim 1, comprising 20. A method for providing a Sustained release of a phar 0.3% ethyl oleate as the fatty acid ester, 0.3% oleic acid as the maceutically active agent through the skin of a Subject, com fatty acid, and 0.75% propylene glycol as the co-solvent, all prising topically administering to the skin of the Subject a by weight of the total weight of the pharmaceutical compo pharmaceutical composition comprising sition. a pharmaceutically active agent comprising one or more 14. The composition according to claim 1, wherein the steroids, pharmaceutically active agent comprises one or more steroids a fatty acid ester, selected from the group consisting of estrogens, anti-estro gens (or SERMs), androgens, anti-androgens, and progestins. Water, 15. The composition according to claim 1, wherein the a C2-C6 monoalcohol and pharmaceutically active agent is selected from one or more of a fatty acid, estradiol and progesterone, and the fatty acid ester is ethyl wherein the weight: weight ratio of the fatty acid ester in the oleate. composition to the total active agent in said composition is at 16. The composition according to claim 1, wherein the least 4:1 fatty acid esteractive agent. pharmaceutically active agent is selected from one or more of 21. The method according claim 20, wherein the fatty acid testosterone and dihydrotestosterone (DHT), and the fatty ester is present in an amount ranging from 0.1% to 20% by acid ester is selected from ethyl oleate and isopropyl weight of the total weight of the pharmaceutical composition. myristate. 22. The method according to claim 20, wherein sustained 17. The composition according to claim 1, wherein the release of the pharmaceutically active agent through the skin active agent is present in an amount ranging from 0.01% to is observed at least 24 hours after said administering. 5% by weight of the total weight of the pharmaceutical com position. c c c c c