Pharmaceutical formulations for delivery of androgen agents and aromatase inhibitors

Abstract

The invention discloses a pharmaceutical preparation which comprises an effective amount of androgen, an effective amount of aromatase inhibitor and a binding agent. Wherein the pharmaceutical formulation is compressed into pellets that, when subcutaneously administered to a patient, provide a sustained release multiphase concentration pattern over time in the blood of the patient. The sustained release multiphase concentration pattern includes: a first time period in which the androgen agent has a first peak concentration in serum and the aromatase inhibitor has an increased concentration in plasma but lower than its first peak concentration in plasma; and a second period of time wherein the androgen agent initially has a reduced serum concentration level and then an increased serum concentration level, the aromatase inhibitor having its first peak concentration in plasma. The invention further discloses application of a combination of the androgen agent and an effective dose of aromatase inhibitor to preparation of a medicine for preventing or treating autoimmune inflammatory mastitis.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of June 3, 2020, application number 202080054087.4, and invention name “Drug preparations and systems for delivering androgens and aromatase inhibitors and methods of use”. Technical Field

[0002] The present disclosure generally relates to pharmaceutical preparations and / or drug delivery systems having novel multiphase release patterns when delivered to warm-blooded animals. The disclosed compositions can be used for warm-blooded animals that wish to increase organ-specific or serum androgen levels without increasing or significantly increasing organ-specific and / or serum estradiol levels. The present disclosure also relates to implants or transdermal patches that provide delivery of multiphase release patterns or pharmacokinetics comprising an effective amount of androgen agents and aromatase inhibitors, which can be used for warm-blooded animals that wish to increase organ-specific or serum androgen levels without increasing or significantly increasing organ-specific and / or serum estradiol levels. Methods of use are also disclosed, including methods associated with tissue-specific autoimmune inflammatory conditions.

[0003] Related references

[0004] Boyd et al., “Evidence for an association between breast tissue firmness and breast cancer risk.” PLoS One 2014 Jul 10; 9(7): e100937

[0005] Cheng Q, Jabbari K, Winkelmaier G, et al. Overexpression of CD36 in mammary fibroblasts inhibits colony growth of breast cancer cell lines. Biochem Biophys Res Commun 2020; 526(1): 41-47.

[0006] Das L, Rai A, Vaiphei K, et al. Idiopathic macromastia: new mechanistic insights involving the paracrine milieu. Endocrine 2019;66(2):166-177.

[0007] Dawson CA, Pal B, Vaillant F, et al. Tissue-resident ductal macrophages survey the mammary epithelium and promote tissue remodeling. Nat Cell Biol 2020; 22(5): 546-558.

[0008] DeFilippis RA, Fordyce C, Patten K, et al. Stress signaling from human mammary epithelial cells contributes to the phenotype of mammographic density. Cancer Res 2014;74(18):5032-44.

[0009] D'Orsi CJ, Sickles EA, Mendelson EB, Morris EA et al (2013).ACR Atlas, the Breast Impact Reporting and Data System. Reston, VA: American College of Radiology.

[0010] Goulabchand R, Hafidi A, Van de Perre P, et al. Mastitis in autoimmune diseases: literature review, diagnostic pathways, and key players in pathophysiology. J Clin Med 2020;9(4).

[0011] Gubbels Bupp MR, Jorgensen TN. Androgen-induced immunosuppression. Front Immunol 2018;9:794.

[0012] Guhl S, Artuc M, Zuberbier T, et al. Testosterone exerts selective anti-inflammatory effects on human skin mast cells in a cell subset-dependent manner. Exp Dermatol 2012; 21(11): 878-80.

[0013] Jeremy Bercoff, “Shear Wave Elastography – A White Paper” SuperSonic Imagine, SA Copyright 2008.

[0014] Liu Y, Sun Y, Zhou Y, et al. Sinomenine hydrochloride inhibits the progression of plasma cell mastitis by regulating the IL-6 / JAK2 / STAT3 pathway. Int Immunopharmacol 2020;81:106025.

[0015] Touraine P, Youssef N, Alyanakian MA, et al. Mastitis macromastia in the context of immune-mediated disease. J Clin Endocrinol Metab 2005; 90(9): 5287-94.

[0016] Uray IP, Liang Y, Hyder SM. Estradiol downregulates CD36 expression in human breast cancer cells. Cancer Lett. 2004; 207(1): 101-7.

[0017] Walecki M, Eisel F, Klug J, et al. Androgen receptor regulates Foxp3 expression in CD4+CD25+Foxp3+ regulatory T cells. Molecular Biology of the Cell, 2015; 10.1091 / mbc.E14-08-1323.

[0018] Wang H, Franco F, Tsui YC, et al. CD36-mediated metabolic adaptation supports the survival and function of regulatory T cells in tumors. Nat Immunol 2020; 21(3): 298-308.

[0019] U.S. Provisional Patent Application No. 62 / 067,297, filed October 22, 2014, and entitled “Methods for Reducing Mammographic Breast Density and / or Breast Cancer Risk.”

[0020] U.S. Provisional Patent Application No. 62 / 324,525, filed April 19, 2016, entitled “METHODS FOR REDUCING MAMMIFIC BREAST DENSITY AND / OR BREAST CANCER RISK”

[0021] U.S. Patent No. 14 / 920,192, filed October 22, 2015, and entitled “Methods for Reducing Mammographic Breast Density and / or Breast Cancer Risk”.

[0022] U.S. Patent No. 15 / 490,309, filed April 18, 2017, and entitled “Methods for Reducing Mammographic Breast Density and / or Breast Cancer Risk”.

[0023] PCT / AU2015 / 000633, filed October 22, 2015, entitled “Methods for reducing mammographic breast density and / or breast cancer risk”.

[0024] Each of these references and PCT / AU2015 / 000633, US Patent Nos. 62 / 067,297, 62 / 324,525, 15 / 490,309, and 14 / 920,192 are incorporated herein by reference in their entirety. Background Art

[0025] Testosterone is essential for regulating immune function in both men and women (Gubbels Bupp, 2018). There are several physiological and disease states where it is desirable to increase tissue levels of 5α dihydrotestosterone (DHT) while simultaneously reducing estradiol levels within the same tissue without causing significant perturbations in serum estradiol levels. DHT is primarily an intra-tissue hormone derived from testosterone that is delivered to the tissue via the circulation. Because the therapeutic window available for administering testosterone without causing androgenic side effects is relatively small, delivering testosterone to tissues in need of treatment in women is problematic. .

[0026] There are also five alpha reductase enzymes that convert testosterone to DHT in most tissues. Therefore, in therapeutic situations where it is desired to increase DHT without increasing estradiol, there is an unmet need and the present disclosure is directed to providing pharmaceutical formulations that aim to inhibit aromatase and induce 5 alpha reductase to achieve a change in the ratio of DHT to estradiol in this tissue.

[0027] Subcutaneous testosterone has been used to provide zero-order kinetic administration of testosterone in both men and women. However, the use of testosterone alone is undesirable in many physiological and disease states because of the presence of high levels of aromatase in the tissue undergoing therapeutic intervention. An example of this is women with high mammographic breast density, in which tissue aromatase levels are very high, which preferentially converts testosterone to estradiol. An estimated 43% of women aged 40 to 75 years in the United States have mammographic breast density (MBD) classified as high, i.e., Breast Imaging Reporting and Data System (Breast Imaging Reporting and Data System) categorized as high. Scores of 3 and 4 (or c and d). The American Cancer Foundation states that this high breast density is a significant risk factor for the development of breast cancer. Traditionally, therapeutic interventions for the perimenopausal transition have been low-dose combination oral contraceptives or continuous estradiol and synthetic progestin delivery systems to protect the uterus from the increased risk of endometrial cancer and unexpected uterine bleeding. These are inappropriate treatments for women with high breast density and / or high breast firmness because they reduce already dangerous levels of testosterone and increase breast density and / or breast firmness. However, these are the current recommendations of the North American Menopause Society and the Australian Menopause Society.

[0028] Another problem in the prior art is breast pain and its treatment. Breast pain is a significant issue in women's health. It is estimated that 45% of women in their 30s say that breast pain affects their quality of life, and 10% say they have experienced breast pain for at least half of their lives. There are few treatments; tamoxifen and aromatase inhibitors have been used as off-label medications for this condition. However, tamoxifen is associated with significant side effects that affect patient compliance, and aromatase inhibitors are contraindicated as single medications in premenopausal women due to the resulting disturbances in the hypothalamic-pituitary-ovarian axis. There is a need in the art for better treatments to reduce breast pain in women.

[0029] The elasticity of breast tissue is believed to be a factor in the development of breast cancer. It has been shown that increased elasticity of breast cells leads to increased mechanotransduction in the cell genome, which may lead to greater malignant transformation. There is a need in the art to provide a composition of a pharmaceutical preparation that reduces mechanotransduction on the cell genome to reduce the risk of malignant transformation.

[0030] A further problem in this area is tissue-specific autoimmune inflammatory conditions, particularly those occurring in the breast. These conditions may be referred to as autoimmune inflammatory mastitis (AIM). The anatomical, histological, and physiological roles of the salivary and mammary glands are similar, as they both belong to the effector sites of the associated mucosal immune system, making them susceptible to autoimmune effects. Autoimmune breast tissue has been shown to have elevated aromatase (which converts androgens to estradiol) and other factors associated with inflammation, such as IGF2, EGFR, TGF-β, PDGFR-α, and β, which have been found to be upregulated, compared to normal breast tissue. AIM is typically treated with extensive immunosuppressive therapy and / or disfiguring surgery, and improved therapies are therefore needed.

[0031] How a pharmaceutical composition is delivered and its effect on the body may be related to the pharmacokinetic and / or pharmacodynamic profiles of the pharmaceutical composition, the drug delivery system, and / or the location of the drug delivery to the subject's body. In a broad sense, pharmacokinetics is a description of the rate and extent of drug absorption, distribution, and / or elimination in the body. In other words, how the body or a part of the body affects the pharmaceutical composition. In a broad sense, pharmacodynamics is a description of how a drug affects the body or a part of the body. Pharmaceutical compositions may have different pharmacokinetic and / or pharmacodynamic profiles. For example, some pharmaceutical compositions may have zero-order, first-order, or second-order kinetics. Kinetics and / or mechanics may affect the efficacy of the pharmaceutical composition and / or the efficacy of the treatment. Therefore, the role of pharmacokinetics and / or pharmacodynamics in certain pharmaceutical compositions or certain drug delivery systems may be related to the results of specific drugs, pharmaceutical compositions, and / or subjects.

[0032] In the past, implants combining testosterone with aromatase inhibitors have been produced, so that the conversion of testosterone to estradiol can be suppressed. However, if the manufacture and concentration of the compound cause the active ingredient to be delivered via zero-order kinetics, the final result is that the serum estradiol level is reduced, which is undesirable in many cases because it causes the potential side effects caused by aromatase inhibitors of estrogen deprivation symptoms and higher sustained levels. There is a continuous demand for pharmaceutical preparations and / or delivery systems for providing DHT and estradiol ratios in high tissues. There is also a continuous demand for the alternative treatment of the treatment of currently available diseases and conditions, and it is beneficial to increase the tissue level of 5α-dihydrotestosterone (DHT) in these diseases and conditions while reducing the estradiol level in the same tissue without causing significant disturbance to the serum estradiol level.

[0033] The present disclosure in one or more embodiments is directed to solving and / or improving one or more shortcomings of the prior art, or at least providing a usable pharmaceutical preparation or therapeutic delivery system, which will become apparent from the disclosure herein. The present disclosure in one or more embodiments may also provide other advantages and / or improvements as discussed herein. Summary of the invention

[0034] In order to obtain a high ratio of DHT to estradiol in tissues, the inventors have recognized that there is an urgent need for a delivery system that delivers testosterone and aromatase inhibitors in a multiphase release mode. In one or more embodiments of the pharmaceutical preparations described herein, an early peak of serum testosterone can be provided, which is rapidly blocked by high levels of aromatase inhibitors, thereby allowing the 5α reductase conversion of testosterone to DHT to be induced. The preparation can then provide a rapid reduction in aromatase inhibitors to ensure that there is no comprehensive blockade of estradiol production and symptomatic reduction of estradiol levels in serum. In tissues where aromatase is overexpressed, using this multiphase release mode, an axis change towards elevated DHT and estradiol levels can be achieved. For example, if the zero-order kinetics of these two components are used, estradiol that will cause adverse effects on women and undesirable long-term exposure of women to aromatase inhibitors higher than the required level for the therapeutic response will not be released.

[0035] In one aspect of the present invention, a pharmaceutical formulation is provided, comprising: an effective amount of an androgen agent, an effective amount of an aromatase inhibitor, and a binding agent; the formulation, when administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by the serum concentration of the androgen agent and the plasma concentration of the aromatase inhibitor; and the sustained-release multiphasic concentration pattern in the subject's serum or plasma comprises:

[0036] a first time period in which the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and

[0037] A second time period wherein the androgen initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

[0038] In another aspect of the present invention, a pharmaceutical formulation is provided, the pharmaceutical formulation comprising: an effective amount of an androgen agent, an effective amount of an aromatase inhibitor, and a binding agent; the formulation, when administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphasic concentration pattern in the subject's serum or plasma comprises:

[0039] a first time period in which the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor concentration in plasma increases but is below its Tmax concentration in plasma;

[0040] a second time period in which the androgen agent initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma;

[0041] a third time period, wherein the androgen agent has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the androgen agent during the third time period; and

[0042] The fourth time period, wherein the serum concentration level of the androgen agent gradually decreases, and the concentration of the aromatase inhibitor in the plasma gradually decreases, and the decrease levels of both are approximately the same as each other.

[0043] In another aspect of the present invention, a pharmaceutical formulation is provided, comprising: an effective amount of an androgen agent, an effective amount of an aromatase inhibitor, and a binding agent; the pharmaceutical formulation is compressed into pellets; the pellets provide a sustained-release multiphasic concentration pattern in the subject's blood over time when subcutaneously administered to a subject, as measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphasic concentration pattern in the subject's serum or plasma comprises:

[0044] a first time period in which the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and

[0045] A second time period wherein the androgen initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

[0046] In another aspect of the present invention, a pharmaceutical formulation is provided, comprising: an effective amount of an androgen agent, an effective amount of an aromatase inhibitor, and a binding agent; the pharmaceutical formulation is compressed into pellets; the pellets provide a sustained-release multiphasic concentration pattern in the subject's blood over time when subcutaneously administered to a subject, as measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphasic concentration pattern in the subject's serum or plasma comprises:

[0047] a first time period in which the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor concentration in plasma increases but is below its Tmax concentration in plasma;

[0048] a second time period in which the androgen agent initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma;

[0049] a third time period, wherein the androgen agent has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the androgen agent during the third time period; and

[0050] The fourth time period, wherein the serum concentration level of the androgen agent gradually decreases, and the concentration of the aromatase inhibitor in the plasma gradually decreases, and the decrease levels of both are approximately the same as each other.

[0051] In at least some embodiments, the pharmaceutical formulation comprises: 60 mg to 120 mg testosterone or an ester thereof, 2 mg to 6 mg aromatase inhibitor, more preferably 4 mg to 6 mg aromatase inhibitor and stearic acid; the pharmaceutical formulation is compressed into pellets having a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm; the pellets provide a sustained-release multiphasic concentration pattern in the subject's blood over time when subcutaneously administered to a subject, as measured by serum concentration of testosterone or an ester thereof and plasma concentration of aromatase inhibitor; and the sustained-release multiphasic concentration pattern comprises:

[0052] a first time period in which testosterone or an ester thereof has a first peak concentration (Tmax) in serum and the concentration of the aromatase inhibitor in plasma increases but is below its Tmax concentration in plasma; and

[0053] A second time period wherein the testosterone or ester thereof initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

[0054] In at least some embodiments, the pharmaceutical formulation comprises: 60 mg to 120 mg testosterone or an ester thereof, 2 mg to 6 mg aromatase inhibitor, more preferably 4 mg to 6 mg aromatase inhibitor and stearic acid; the pharmaceutical formulation is compressed into pellets having a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm; the pellets provide a sustained-release multiphasic concentration pattern in the subject's blood over time when subcutaneously administered to a subject, as measured by serum concentration of testosterone or an ester thereof and plasma concentration of aromatase inhibitor; and the sustained-release multiphasic concentration pattern comprises:

[0055] a first time period in which testosterone or an ester thereof has a first peak concentration (Tmax) in serum and the concentration of the aromatase inhibitor in plasma increases but is below its Tmax concentration in plasma;

[0056] a second time period in which the testosterone or its ester initially has a decreased serum concentration level and then has an increased serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma;

[0057] a third time period, wherein testosterone or its ester has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of testosterone or its ester during the third time period; and

[0058] A fourth time period, wherein the serum concentration level of testosterone or its ester gradually decreases, and the concentration of the aromatase inhibitor in plasma gradually decreases, and the decrease levels of the two are approximately the same as each other.

[0059] In certain exemplary embodiments, the aromatase inhibitor does not exhibit zero order release in the pharmaceutical formulation during the first time period.

[0060] In certain exemplary embodiments, the aromatase inhibitor exhibits a first order release in the pharmaceutical formulation during the first time period.

[0061] In certain exemplary embodiments, the aromatase inhibitor does not exhibit zero order release in the pharmaceutical formulation during the second time period.

[0062] In certain exemplary embodiments, the androgenic agent (eg, testosterone or an ester thereof) in the pharmaceutical formulation during the second time period does not exhibit zero order release.

[0063] In certain exemplary embodiments, the aromatase inhibitor does not exhibit a zero order release of the drug during the third time period.

[0064] In certain exemplary embodiments, the androgen agent does not exhibit zero order release in the pharmaceutical formulation during the third time period.

[0065] In certain exemplary embodiments, the aromatase inhibitor exhibits a first order release in the pharmaceutical formulation during the third time period.

[0066] In certain exemplary embodiments, the androgen agent exhibits a first order release in the pharmaceutical formulation during the third time period.

[0067] In certain exemplary embodiments, the aromatase inhibitor does not exhibit zero order release in the pharmaceutical formulation during the fourth time period.

[0068] In certain exemplary embodiments, the androgen agent does not exhibit zero order release in the pharmaceutical formulation during the fourth time period.

[0069] In certain exemplary embodiments, the aromatase inhibitor exhibits a first order release in the pharmaceutical formulation during the fourth time period.

[0070] In certain exemplary embodiments, the androgen agent exhibits a first order release in the pharmaceutical formulation during the fourth time period.

[0071] In some exemplary embodiments of the pharmaceutical preparation, the first time period ends immediately after the androgen has the first peak concentration (Tmax) in the serum. In some exemplary embodiments of the pharmaceutical preparation, the first time period ends at 5 hours to 14 hours. In some exemplary embodiments of the pharmaceutical preparation, the first time period ends at 5.5 hours to 13 hours.

[0072] In certain exemplary embodiments of the pharmaceutical formulation, the second time period ends immediately after the aromatase inhibitor has its Tmax. In certain exemplary embodiments of the pharmaceutical formulation, the second time period ends between 23 hours and 80 hours.

[0073] In addition, certain embodiments relate to a pharmaceutical formulation comprising: about 80 mg of testosterone or an ester thereof, about 4 mg of anastrozole, and about 2 mg of stearic acid; the pharmaceutical formulation is compressed into pellets having a diameter of 4.4 mm to 4.6 mm and a length of 4 mm to 7 mm; the pellets provide a sustained-release multiphasic concentration pattern in the subject's blood over time when subcutaneously administered to a subject, as measured by serum concentration of testosterone or an ester thereof and plasma concentration of anastrozole; the sustained-release multiphasic concentration pattern comprising: a first time period, in which testosterone or an ester thereof has a first peak concentration (Tmax) in serum and anastrozole in plasma The invention discloses a method for using a pharmaceutical preparation in which the serum concentration level of testosterone or its ester is gradually reduced and the concentration of anastrozole in plasma is gradually reduced, and the concentration of anastrozole in plasma is gradually reduced, and the reduction level of the two is approximately the same as each other. A method for using the pharmaceutical preparation is also disclosed.

[0074] Further embodiments relate to a pharmaceutical formulation comprising: an effective amount of an androgen, an effective amount of an aromatase inhibitor, and a binding agent; the pharmaceutical formulation is compressed into pellets; the pellets provide a sustained-release multiphasic concentration pattern in the subject's blood over time when subcutaneously administered to a subject, as measured by serum concentration of the androgen and plasma concentration of the aromatase inhibitor; and the sustained-release multiphasic concentration pattern in the subject's serum or plasma includes: a first time period, wherein the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor does not have its Tmax concentration in plasma; a second time period, wherein the androgen has a decreased serum concentration level and then has an increased serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma; a third time period, wherein the androgen has a second peak concentration in serum below its Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the androgen within the third time period; and a fourth time period, wherein the androgen has a decreased serum concentration level and the aromatase inhibitor has a decreased concentration in plasma.

[0075] Further embodiments are directed to methods of reducing mammographic breast density as measured by AVBD and / or VBD% in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0076] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing VBD% in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0077] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing AVBD in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0078] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing AVBD and VBD% in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0079] Further embodiments relate to the use of a pharmaceutical formulation in a method of treating mammographic breast density as measured by AABD and / or ABD % in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0080] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing ABD % in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0081] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing AABD in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0082] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing AABD and ABD % in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0083] Further embodiments are directed to the use of a pharmaceutical formulation in a method of reducing mammographic breast density in a patient having breasts with a mammographic breast density of 7.5% or greater, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0084] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing VBD% in a patient having a breast with 7.5% or greater VBD% comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0085] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing ABD% in a patient having a breast with 7.5% or greater ABD% comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0086] Further embodiments relate to pharmaceutical preparations for treating Use in a method of treating a patient having a mammographic breast density score of 3 or 4 (or c or d) comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0087] Further embodiments relate to pharmaceutical preparations that have Use in a method of treating a patient having a mammographic breast density score of 3 or 4 (or c or d) comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0088] Further embodiments relate to the use of a pharmaceutical formulation in a method of inducing breast involution in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0089] Further embodiments relate to the use of a pharmaceutical formulation in a method of inducing net cell death exceeding proliferation in the breast of a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0090] Further embodiments relate to the use of a pharmaceutical formulation in a method of inducing net extracellular matrix degradation exceeding extracellular matrix development in the breast of a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0091] Further embodiments relate to the use of a pharmaceutical formulation in a method of reversing breast cell count and mammographic breast density in a perimenopausal patient, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0092] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing mammographic breast density and perimenopausal symptoms in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0093] Further embodiments relate to the use of the pharmaceutical formulation in a method useful in premenopausal and / or perimenopausal women for reducing the risk of breast cancer while substantially not causing perturbations of the hypothalamic-pituitary axis and / or other endocrine axes such as the adrenal glands and / or ovaries.

[0094] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing breast firmness in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0095] Further embodiments relate to the use of a pharmaceutical formulation in a method of alleviating breast pain in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0096] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing breast elasticity in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0097] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing mechanical transduction on the cellular genome to reduce the risk of malignant transformation in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0098] Further embodiments relate to the use of a pharmaceutical formulation in a method of increasing the fibroglandular to adipose tissue ratio in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0099] Further embodiments relate to the use of a pharmaceutical formulation in a method of increasing CD36 in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0100] Further embodiments relate to the use of a pharmaceutical formulation in a method of stabilizing and / or increasing the level of androgen receptor expression in breast tissue of a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0101] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing and / or treating macromastia in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0102] Further embodiments relate to the use of a pharmaceutical formulation in a method of increasing GCDFP15 in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0103] Further embodiments relate to use of a pharmaceutical formulation in a method of reducing breast pain associated with a mammographic image taken in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0104] Further embodiments relate to the use of a pharmaceutical formulation in a method of increasing mammographic sensitivity in a patient, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0105] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing ABD % and / or AABD in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0106] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing BPE in an MRI image of a patient, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0107] Further embodiments relate to the use of a pharmaceutical formulation in a method of reducing the size and / or number of cysts in a patient in need thereof, comprising administering to the patient a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0108] Additionally, further embodiments relate to preventing or treating autoimmune inflammatory mastitis in a patient in need thereof.

[0109] In particular, in another aspect of the present invention, there is provided a method of preventing or treating autoimmune inflammatory mastitis in a patient in need thereof, comprising administering to the patient 1) an effective amount of an androgen agent and 2) an effective amount of an aromatase inhibitor.

[0110] In this embodiment, the androgen and aromatase inhibitor can be administered to the patient in the same pharmaceutical formulation or in separate formulations. Therefore, the present disclosure extends to all combined treatments for autoimmune inflammatory mastitis in patients. Combined treatment means that the androgen and aromatase inhibitor can be administered together simultaneously or sequentially through the same or different routes (which can be determined by a doctor or nursing staff), so that the androgen and aromatase inhibitor play their respective therapeutic effects in overlapping therapeutic windows.

[0111] Typically, in at least some embodiments as described herein for treating autoimmune inflammatory mastitis, the androgenic agent and the aromatase inhibitor are administered in the same or different sustained release pharmaceutical formulations.

[0112] In particularly preferred embodiments, one or more sustained release pharmaceutical formulations are used to deliver the androgen agent and the aromatase inhibitor to the patient subcutaneously, for example in the form of one or more solid dosage forms such as one or more pellets.

[0113] Most typically, androgen and aromatase inhibitor are provided with the same sustained release pharmaceutical formulation.Sustained release formulations can be, for example, pharmaceutical formulations as described herein for providing a sustained release multiphasic concentration pattern over time in subject's blood, as measured by the serum concentration of androgen and the plasma concentration of aromatase inhibitor.

[0114] Thus, further embodiments relate to the use of a pharmaceutical formulation in a method of treating autoimmune inflammatory mastitis in a patient in need thereof, comprising administering to the patient the pharmaceutical formulation in the form of a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0115] The autoimmune inflammatory mastitis of a patient treated according to the present disclosure can, for example, be selected from the group consisting of idiopathic inflammatory macromastitis, plasma cell mastitis, granulomatous mastitis, and combinations thereof.

[0116] Accordingly, further embodiments of the present disclosure relate to the use of a pharmaceutical formulation in a method of treating idiopathic inflammatory macromastia associated with an autoimmune disease in a patient in need thereof, comprising administering to the patient the pharmaceutical formulation in the form of a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0117] Further embodiments of the present disclosure are directed to the use of a pharmaceutical formulation in a method of treating plasma cell mastitis in a patient in need thereof, comprising administering to the patient the pharmaceutical formulation in the form of a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0118] Further embodiments of the present disclosure are directed to the use of a pharmaceutical formulation in a method of treating granulomatous mastitis in a patient in need thereof, comprising administering to the patient the pharmaceutical formulation in the form of a subcutaneous pellet comprising: i) an effective amount of an androgenic agent; and ii) an effective amount of an aromatase inhibitor.

[0119] The invention further extends to the use of an androgenic agent in combination with an effective amount of an aromatase inhibitor in the manufacture of a medicament for preventing or treating autoimmune inflammatory mastitis in a patient in need thereof.

[0120] In yet another embodiment, there is provided a use of an aromatase inhibitor in combination with an effective amount of an androgen agent for the preparation of a medicament for preventing or treating autoimmune inflammatory mastitis in a patient in need thereof.

[0121] Typically, the androgenic agent administered or included in the pharmaceutical formulations for treating autoimmune inflammatory mastitis (AIM) or AIM conditions as described herein is testosterone or a pharmaceutically acceptable salt or ester thereof. Aromatase inhibitors may also be used in the form of their pharmaceutically acceptable salts or esters.

[0122] As is apparent from the above, the present disclosure explicitly extends to methods for administering an effective amount of a pharmaceutical formulation as described herein to a subject for the purposes and / or treatments described herein. In addition, in certain embodiments, there is provided the use of androgens and aromatase inhibitors in the preparation of medicaments as described herein. The overview provided above is not intended to limit the embodiments disclosed herein and other embodiments disclosed in this specification. In addition, the limitations of one embodiment may be combined with the limitations of other embodiments to form additional embodiments.

[0123] definition

[0124] Unless otherwise defined below, the terms used herein are as commonly used in the art:

[0125] The term "absolute area of ​​breast density" (AABD) refers to the measurement of the surface area of ​​fibroglandular tissue in a subject's mammogram in square centimeters. This can be measured, for example, using the CUMULUS software algorithm or visual inspection of the mammogram. Several other tests can be used to measure AABD, including but not limited to VOLPARA, QUANTRA, and methods that account for the surface area of ​​fibroglandular tissue in a mammogram.

[0126] The term "androgen" refers to an agent that increases androgen activity and / or synthesis. For example, an androgen can be a steroid hormone that binds to its intracellular mediator androgen receptor with high affinity (within the range of pM or nM) and specificity, resulting in a change in receptor conformation, thereby allowing cofactor inclusion, nuclear transport and / or stimulating transactivation activity. Thus regulating the expression of a target gene. For example, an androgen can be an androgen, such as an androgen selected from the group consisting of: testosterone, methyltestosterone and dehydroepiandrosterone. The combination of these androgen agents can also be considered. In addition to the pharmaceutically acceptable ester of testosterone, esters can include but are not limited to enanthate, propionate, cypionate, phenylacetate, acetate, isobutyrate, cyclobutyl, enanthate, decanoate, undecanoate, decyl ester and / or isodecyl ester.

[0127] The term "breast area density percentage" (ABD%) refers to the proportion or percentage of fibroglandular (dense) tissue relative to the total surface area of ​​the breast on a mammogram. This can be measured, for example, using the CUMULUS software algorithm or visual inspection of the mammogram. Several other tests that can be used to measure ABD% include, but are not limited to, VOLPARA, QUANTRA, and methods that account for the surface area of ​​fibroglandular tissue in the mammogram.

[0128] The term "aromatase inhibitor" refers to a compound, hormone or polypeptide that blocks and / or inhibits the activity of aromatase, an enzyme that converts androgens into estrogens. For example, the aromatase inhibitor can be selected from the group consisting of anastrozole, exemestane and letrozole.

[0129] The term "autoimmune inflammatory mastitis (AIM)" refers to a condition in which non-infectious, non-lactational inflammation of the breast tissue occurs due to an autoimmune response to components within the breast tissue.

[0130] The term "breast cancer" refers to a malignant proliferation of the epithelial cells lining the breast ducts or lobules.

[0131] The term "breast elasticity" refers to a measure of the pressure required to achieve a given partial deformation of a breast or breast portion. For example, elasticity = pressure / fractional change in breast radius, where pressure is measured in, for example, kilopascals, and fractional change in breast radius = (R1-R2) / R1, where R1 is the uncompressed radius, and R2 is the compressed radius. Another example may be to directly measure the elasticity of breast tissue by applying shear waves through the breast tissue, for example, using a microscope utilizing SuperSonic Imagine's ShearWave TM SuperSonic Imagine Aixplorer by Elastography TM to measure.

[0132] The term "breast firmness" in its broadest sense refers to a measure of the tolerance of the breast to deformation. Factors that may affect the degree of breast firmness include, but are not limited to, physical forces generated by interactions between cells and between cells and the extracellular matrix, the number of cells present in the breast and the degree of collagen, the degree of fluid retention in the breast, the degree of proteoglycan expression, and / or combinations thereof. An example of measuring breast firmness includes using the formula force / deformation (dN / cm), where dN represents ten Newtons and cm represents centimeters, where the deformation can be determined as the difference between the radius of the semicircle of the mammographic area and the radius of the volume hemisphere, and the pressure is recorded by a mammogram, such as a digital mammogram. For example, according to Boyd et al., the deformation can be determined as the difference between R1-R2, where R1 is the uncompressed radius and R2 is the compressed radius (Boyd et al., "Evidence that breast tissue firmness is associated with breast cancer risk" PLoS One 2014 Jul 10; 9(7): e100937).

[0133] The term "breast tissue" refers to the collection of epithelial cells, stromal cells, extracellular matrix, and / or migratory cells located in or near the breast.

[0134] The term "effective amount" or "pharmaceutically effective amount" of an agent or compound refers to an amount of an agent or compound that is sufficient to provide the desired therapeutic effect and is non-toxic, has an acceptable non-toxicity profile, and / or an acceptable side effect profile. The required amount may vary from patient to patient depending on, for example, the patient's age, general condition, severity of the condition being treated, the specific agent or compound being administered, and one or more combinations of these factors. One skilled in the art can determine the generally appropriate "effective amount" in an individual case by reference to relevant textbooks and literature and / or using routine experimentation.

[0135] The term "mammographic breast density" or "MBD" refers to a qualitative estimate of the proportion or percentage of radiopaque or fibroglandular ("dense") elements / tissue in the breast relative to total breast area (measured by 2-D) or volume (measured by 3-D). Mammographic breast density (MBD) is a qualitative or quantitative estimate of the amount of fibroglandular tissue (FGT) within the breast. It can be the absolute amount of FGT or the relative amount of FGT to the amount of non-FGT (primarily fat or adipose). It can be measured in terms of surface area (cm 2 ) (i.e. AABD) or volume (cm 3The relative amount of FGT can be an estimate of the surface area of ​​the FGT relative to the non-FGT as a percentage (i.e., ABD%) or an estimate of the volume of the FGT relative to the volume of the breast (i.e., VBD%). Another approach can be to determine the absolute volumetric breast density (i.e., AVBD), which can be the measured volume of fibroglandular tissue in a subject's breast in cubic centimeters. Mammographic breast density can be determined by a variety of methods, including, but not limited to, mammography, digital mammography, magnetic resonance imaging (MRI), ultrasound, digital breast tomosynthesis (DBT), palpation tissue imaging quantification (VTIQ), and combinations thereof. Breast density can be determined by 2-D measurement and / or using Density classification qualitatively evaluates MBD. Density classifications of 1 (or a) being the least dense, and 4 (or d) being the most dense. MBD can also be assessed qualitatively and / or quantitatively by 3-D measurements and / or using volumetric measurements of the breast, such as determining volumetric breast density, which is the ratio of fibroglandular (dense) tissue in the breast relative to the total volume of tissue (e.g., fibroglandular (dense) tissue and fat in the breast). Another approach is to determine absolute volumetric breast density (i.e., AVBD), which is the measured volume of fibroglandular tissue in a subject's breast in cubic centimeters. Assessing MBD by 3-D measurements can also account for the heterogeneity of dense tissue within the breast. Several tests that can be used to measure MBD include, but are not limited to, VOLPARA, QUANTRA, CUMULUS, and VIBRATION that considers the volume of fibroglandular tissue (cm 3 ) method.

[0136] The term "patient" or "subject" refers to animals, including human species, that can be treated with the compositions, methods, and kits of the present disclosure. Unless one sex is specifically indicated or one sex is clear from the context, the term "patient" is intended to refer to both males and females. The term "patient" may also refer to a female-to-male transgender person.

[0137] The term "perimenopause" or "menopause transition" refers to a period of time near menopause during which a woman's body makes its natural transition to permanent infertility (menopause). Women may begin perimenopause at different ages, and in their 40s or even as early as their mid-30s, may notice signs of progression to menopause, such as irregular periods. During perimenopause, estrogen levels may rise and fall unevenly, menstrual cycles may lengthen or shorten, and menstrual cycles in which the ovaries do not release eggs (ovulation) may begin. During perimenopause, other menopausal-like symptoms may be experienced, including, but not limited to, hot flashes, sleep problems, and / or vaginal dryness.

[0138] The term "perimenopausal symptoms" should be understood to include, but are not limited to, menstrual irregularities; hot flashes and sleeping problems; mood changes; mood swings; irritability; depression; vaginal dryness; urinary tract infections or vaginal infections; urinary incontinence; decreased fertility; changes in sexual arousal or desire; bone loss; bone fragility; osteoporosis; or changes in cholesterol levels, such as increased low-density lipoprotein (LDL) cholesterol or decreased high-density lipoprotein (HDL) cholesterol, and combinations thereof.

[0139] The term "pharmaceutically acceptable" refers to those compounds, agents, materials, compositions, excipients, and / or dosage forms which are, within the scope of sound medical judgment, suitable for contact with human and / or animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable ratio of benefits and / or risks.

[0140] The term "pellet" refers to a solid dosage implant comprising an effective amount of androgen and an effective amount of an aromatase inhibitor. The pellet or implant may have different suitable shapes and sizes, such as spherical, cylindrical, rectangular, square or a combination thereof. It may have angled edges or rounded edges. In certain embodiments, the pellet or implant may be compressed.

[0141] The term "pharmaceutical preparation" refers to a preparation comprising an effective amount of androgen and an effective amount of an aromatase inhibitor, which, when applied to a warm-blooded subject, provides a sustained release multiphasic concentration pattern in the subject's blood over time, as measured by androgen serum concentration and aromatase inhibitor plasma concentration. There may be at least 2, 3 or 4 different time periods. These time periods vary according to one or more of the following: preparation, delivery system, drug concentration, and individual physiological changes during the completion or substantial completion of preparation absorption. Typically, the absorption of the preparation is no less than 3 months and no more than 5 months. In certain embodiments, the preparation can be delivered by a transdermal patch. In certain embodiments, the preparation can be delivered by a solid form such as a subcutaneous pellet. In certain embodiments, the preparation can be delivered by a solid form such as a compressed subcutaneous pellet.

[0142] The term "postmenopausal woman" should be understood to include not only older women who have passed menopause, but also women who have had their ovaries removed or destroyed by other means or whose estrogen production has been suppressed for some other reason, such as women undergoing long-term corticosteroid administration, suffering from Cushing's syndrome, or having gonadal dysgenesis.

[0143] The term "subject" is an animal, including human species, that can be treated with the compositions, methods and kits of the present disclosure. Unless one sex is specifically indicated or one sex is clear from the context, the term "subject" is intended to refer to both males and females. The term "subject" may also refer to a female to male transgender person.

[0144] As used herein, the terms "treatment" or "therapy" include preventative (eg, prophylactic) treatment and / or palliative treatment, and as used herein, "treatment" refers to the act of providing preventative and / or palliative treatment.

[0145] The term "Volume Breast Density Percent (VBD%)" refers to the ratio or percentage of the volume of fibroglandular (dense) tissue relative to the total volume of tissue in the breast. This can be done, for example, using Volpara Solution TM Software algorithm measurement. In Volpara Solution TM In the software algorithm, VBD% is referred to as volumetric breast density percentage. Several other tests that can be used to measure VBD% include, but are not limited to, QUANTRA, CUMULUS, and methods that take into account the volume of fibroglandular tissue. BRIEF DESCRIPTION OF THE DRAWINGS

[0146] In order to better understand the present disclosure, and to more clearly show how it can be implemented according to one or more embodiments of the present disclosure, reference will now be made to the accompanying drawings by way of example. A sustained release concentration profile over time in the blood of a subject is provided, as measured by serum concentration of testosterone or its ester and plasma concentration of an aromatase inhibitor.

[0147] Figure 1 Data for serum concentrations of androgens and plasma concentrations of aromatase inhibitors over 84 days are shown, according to certain exemplary embodiments.

[0148] Figure 2 Shows Figure 1 , but expanded to further account for the first 42 days.

[0149] Figure 3 Shown are testosterone serum concentrations for 11 subjects up to 7 days post-dosing according to certain exemplary embodiments.

[0150] Figure 4 Shown are testosterone serum concentrations for 11 subjects up to 85 days after dosing, according to certain exemplary embodiments.

[0151] Figure 5 Shown are plasma concentrations of anastrozole in 11 subjects over 336 hours post-dose, according to certain exemplary embodiments.

[0152] Figure 6 Shown are plasma concentrations of anastrozole in 11 subjects over 1680 hours post-dose, according to certain exemplary embodiments.

[0153] Figure 7A-7BThe absorption rate of anastrozole according to certain embodiments is illustrated.

[0154] Figures 8A-8B Anastrozole plasma concentrations following implantation according to certain embodiments are described.

[0155] Fig. 9 The observed frequencies after implantation in Example 3 are shown.

[0156] Fig.10 An example flow chart illustrating the model tested in Example 3.

[0157] Fig.11 The final structural model used in Example 3 is shown.

[0158] Figures 12A-12B The population and individual predictions used in Example 3 are shown.

[0159] Fig.13 Eleven individual graphs illustrating observed and predicted plasma concentrations of anastrozole according to certain embodiments.

[0160] Fig.14 The visual predictive check (VPC) of post-implantation anastrozole concentration used in Example 3 is shown.

[0161] Figures 15A-15B The one-compartment model and the two-compartment model used in Example 3 are shown.

[0162] Fig.16 Predictions for patient 10 from several different input models used in Example 3 are shown.

[0163] Fig.17 Schematic illustration of the effects of the androgenic agents testosterone and aromatase inhibitor (T+Ai) on breast tissue immunoreactivity.

[0164] Fig.18 Shown are the results of western blot analysis of CD36 protein in three explant samples of normal breast tissue obtained from three perimenopausal women at baseline and after 24 hours of culture.

[0165] Fig.19 A is a mammographic image before (left) and after (right) treatment according to the present disclosure, showing a significant reduction in breast size and density.

[0166] Fig.19 B is Fig.19 A. Volpara Solution before treatment (left) and after treatment (right) TM Breast volume and density were measured, which showed a reduction in breast volume (58%) back to pre-morbid size and a reduction in fibroglandular tissue (71%).

[0167] Fig.19 C is Fig.19 Magnetic resonance images (MRI) of a patient's breast before treatment (left) and three years after treatment, showing reversal of the extreme background parenchymal enhancement on MRI over a three-year period after treatment was stopped.

[0168] Fig. 20 A is a mammographic image of another patient before (left) and after (right) treatment according to the present disclosure, showing a significant reduction in breast size and density.

[0169] Fig. 20 B is Fig. 20 A. Volpara Solution before treatment (left) and after treatment (right) TM Breast volume and density were measured, which showed a decrease in breast volume (10%) back to pre-morbid size and a decrease in fibroglandular volume (41%).

[0170] Fig.21 A shows mammographic images of another patient before (left) and after (right) treatment according to the present disclosure, showing a significant reduction in breast size and density.

[0171] Fig.21 B is Fig.21 A. Volpara Solution before treatment (left) and after treatment (right) TM Breast volume and density were measured, which showed a decrease in breast volume (32%) back to pre-morbid size and a decrease in fibroglandular volume (52%).

[0172] Fig. 22 A is a magnetic resonance image (MRI) of the breast of another patient before (left) and after (right) treatment according to the present disclosure, showing complete reversal of the extreme background parenchymal enhancement on MRI.

[0173] Fig. 22 B is Fig. 22 Mammographic images of patient B's breasts before (left) and after (right) treatment, showing a reduction in breast size and density.

[0174] Fig. 22 C is Fig. 22 Volpara Solution before (left) and after (right) treatment of patient B TM Breast volume and density were measured, which showed a decrease in breast volume (23%) back to pre-morbid size and a decrease in fibroglandular volume (36%).

[0175] Fig.23A shows extreme breast swelling in another patient as confirmed by mammography prior to therapeutic treatment according to the present disclosure.

[0176] Fig.23 B shows a magnetic resonance imaging (MRI) of a highly metabolically active breast with very little fat, such as Fig.23 The extreme background parenchymal enhancement in the breast of patient A is shown.

[0177] Fig.24 A shows a magnetic resonance image (MRI) of another patient with reactive axillary lymphadenopathy with granulomatous formation in the retroareolar space.

[0178] Fig.24 B is Fig.24 MRI image of the breast of patient A showing regression of granulomas and reduction in axillary lymph node size after treatment according to the present disclosure.

[0179] In view of the following detailed description and appended claims, those skilled in the art will appreciate that various alterations and / or modifications may be made to the present disclosure without departing from the scope of the invention as claimed. DETAILED DESCRIPTION

[0180] The following description is provided with respect to several embodiments that may share common characteristics and features. It should be understood that one or more features of one embodiment may be combined with one or more features of other embodiments. In addition, a single feature or a combination of features in certain embodiments may constitute additional embodiments. The specific structural and functional details disclosed herein should not be interpreted as limiting, but only as a representative basis for teaching those skilled in the art to use the disclosed embodiments and variations of those embodiments in various ways.

[0181] The subject headings used in the detailed description are included only for the convenience of the reader and should not be used to limit the subject matter found in the entire disclosure or claims. The subject headings should not be used to interpret the scope of the claims or the limitations of the claims.

[0182] The present disclosure relates at least in part to providing a drug delivery system having a multi-phase sustained release pattern, which comprises an androgen and an aromatase inhibitor in a compressed pellet, unexpectedly overcoming the problem of obtaining a high intra-tissue DHT to estradiol ratio when delivered subcutaneously to a subject. The desired effect is to rapidly block the early peak of serum testosterone by a high level of aromatase inhibitor, thereby inducing the 5α-reductase conversion of testosterone to DHT. The rapid reduction of the aromatase inhibitor is then desired to ensure that there is no blockage or substantial blockage of overall estradiol production and a symptomatic reduction in estradiol levels in serum. By utilizing certain exemplary embodiments that exhibit this multi-phase sustained release pattern, a change in the axis toward high DHT to estradiol levels in tissues where aromatase overexpression is present can be achieved.

[0183] There are several examples where aromatase overexpression leads to disease processes where women and men may benefit from this combination of testosterone and aromatase inhibitors. Thus, in these tissues, where there may be overexpression of aromatase and coexpression of 5α-reductase, this enzymatic combination may be pharmacologically manipulated to achieve an orderly shift toward altered DHT / estradiol ratios. These disease conditions include (but are not exclusive to) high mammographic breast density, breast pain, endometriosis, gynecomastia, perimenopausal and premenstrual tissue aberrations, autoimmune inflammatory mastitis (including conditions such as idiopathic inflammatory macromastia, plasma cell mastitis, and granulomatous mastitis).

[0184] Figure 2 An example of a testosterone and aromatase inhibitor combination is illustrated, as well as the effects of this paradigm on several subjects (N=11) who have been administered a subcutaneous pellet regimen.

[0185] In this example, Phase I shows a rapid increase in both serum testosterone and plasma anastrozole in the subject. This increase results in rapid tissue induction of 5α-reductase and severe inhibition of aromatase. High tissue levels of testosterone are rapidly converted to DHT, and conversion to estradiol is completely blocked or substantially blocked. In this example, the first phase lasts approximately 8 days, as shown by the vertical line separating Phase 1 from Phase 2. The time period of Phase 1 can vary depending on a variety of factors, including, but not limited to, the biology of the particular subject, the formulation of the pellets, and / or the manufacturing parameters used to manufacture the pellets.

[0186] In the example, the Phase II time period lasts for approximately 14 days. In Phase 2, it is desirable to re-regulate the enzymatic system to induce sustained high intra-tissue anastrozole levels, which continue to allow the induced 5α-reductase to work unopposed or substantially unopposed to perform the aromatase conversion of testosterone and estradiol. This re-regulation results in a drop in serum testosterone, avoiding excessive androgenic side effects and continued use of 5α-reductase. In this example, Phase II lasts for approximately 14 days, as indicated by the vertical line separating Phase 2 from Phase 3. The time period for Phase 2 may vary depending on a variety of factors, including, but not limited to, the biology of the particular subject, the formulation of the pellets, and / or the manufacturing parameters used to manufacture the pellets.

[0187] In this example, the levels of anastrozole in Phase III decline rapidly, allowing some testosterone to be converted to estradiol and avoiding heavy use and the exhaustion symptoms that might be caused by sustained high levels of anastrozole observed in a zero-order release pattern. During this time, the induced 5-alpha reductase continues to convert elevated testosterone to DHT within the target tissue until the implant runs out of substrate and is replaced. In this example, this Phase III lasts at least until the pellet is exhausted. Figure 1 As shown, the subjects were followed for an additional 70 days. The time period for Phase 3 may vary depending on a number of factors, including, but not limited to, the biology of the particular subject, the formulation of the pellets, and / or the manufacturing parameters used to manufacture the pellets.

[0188] Certain embodiments can be used to treat extreme mammographic breast density. Serum reproductive hormone levels are weakly correlated with levels in tissues because a large amount is metabolized to inactive or active metabolites of prohormones (such as testosterone). In certain disease states, such as extreme mammographic breast density, there is an overexpression of enzymes known to convert testosterone to estrogen or dihydrotestosterone (10 times more potent than testosterone as an androgen). By changing the ratio of androgens to estrogens in high mammographic breast density, it is possible to reduce this density and therefore reduce the incidence of breast cancer. The reduction in density is also applicable to situations such as gynecomastia where there is excess estrogen / androgen in the tissue. In order to promote the change in the ratio of androgens to estrogens, a subcutaneous pellet is designed to promote a multi-phase sustained release mode. When anastrozole and testosterone are combined in a specific manner and compressed into a pellet that has been specifically compressed, the resulting pharmacokinetics promote this multi-phase sustained release of the two active ingredients. Likewise, as discussed elsewhere, it is desirable that the early increase of anastrozole obtain the minimum concentration required to promote the blockade or substantial blockade of aromatase in tissues expressing high levels of aromatase at about the time of peak testosterone levels. The early increase of anastrozole promotes the conversion of testosterone to dihydrotestosterone by 5α reduction, and promotes an environment of excess androgen relative to estrogen. Once this initial high level of anastrozole is obtained, a significantly low level is maintained to avoid causing changes in hypothalamic-pituitary function or induction of systemic aromatase inhibition as seen in higher concentrations of aromatase inhibitors when, for example, oral aromatase inhibitors. This then allows testosterone to slowly enter the tissue, where 5α reductase is induced in the somatic cell system to continue to perturb the antigenic environment. This unique multi-phase sustained release pattern can be used to change tissue hormone levels without causing significant physiological changes in other serum reproductive hormone levels and potential adverse side effects resulting therefrom.

[0189] Certain embodiments can be used to treat one or more of the following: high breast density and breast firmness. High breast density and / or breast firmness are not normal, but rather pathological, and can sometimes be addressed with an effective amount of an androgen agent and / or an effective amount of an aromatase inhibitor, rather than with lifestyle changes (e.g., diet and exercise), which have not been shown to be successful in premenopausal, perimenopausal, and / or postmenopausal women.

[0190] Certain embodiments relate to a pharmaceutical formulation that can be used in a method for providing an individualized reduction of one or more of the following: mammographic breast density and breast firmness of a patient in need thereof, comprising (i) determining the patient's MBD and / or breast firmness; (ii) optionally, measuring the patient's free androgen index and / or the change in the patient's free androgen index over a period of at least one month; (iii) determining an adjusted dose of an androgen agent and an aromatase inhibitor taking into account the patient's weight, total body fat, MBD, age, and free androgen index; and (iv) administering the adjusted dose to the patient. In certain embodiments, measuring the patient's free androgen index and / or the change in the patient's free androgen index over a period of at least one month may include collecting a blood sample and measuring the amount of free androgen agent (or testosterone) in the patient's serum.

[0191] Certain embodiments relate to a pharmaceutical formulation that can be used in a method for providing personalized reduction of one or more of the following: VBD%, AVBD, and breast firmness of a patient in need thereof, comprising (i) determining the patient's VBD% and / or AVBD and / or breast firmness; (ii) optionally, measuring the patient's free androgen index and / or the change in the patient's free androgen index over a period of at least one month; (iii) determining an adjusted dose of an androgen agent and / or an aromatase inhibitor taking into account the patient's weight, total body fat, VBD% and / or AVBD, age, and free androgen index; and (iv) administering the adjusted dose to the patient. In certain embodiments, measuring the patient's free androgen index and / or the change in the patient's free androgen index over a period of at least one month may include collecting a blood sample and measuring the amount of free androgen agent (or testosterone) in the patient's serum.

[0192] Certain embodiments relate to a pharmaceutical formulation that can be used in a method for providing personalized reduction of one or more of the following: ABD%, AABD, and breast firmness in a patient in need thereof, comprising (i) determining the patient's VBD% and / or AVBD and / or breast firmness; (ii) optionally, measuring the patient's free androgen index and / or the change in the patient's free androgen index over a period of at least one month; (iii) determining an adjusted dose of an androgen agent and / or an aromatase inhibitor taking into account the patient's weight, total body fat, ABD% and / or AABD, age, and free androgen index; and (iv) administering the adjusted dose to the patient. In certain embodiments, measuring the patient's free androgen index and / or the change in the patient's free androgen index over a period of at least one month may include collecting a blood sample and measuring the amount of free androgen agent (or testosterone) in the patient's serum.

[0193] Certain embodiments relate to pharmaceutical formulations useful for reducing one or more of: mammographic breast density and breast firmness in a patient in need thereof, wherein the pharmaceutical formulation increases the sensitivity of breast imaging detection by mammography, digital mammography, magnetic resonance imaging (MRI), ultrasound, digital breast tomosynthesis (DBT), palpation tissue imaging quantification (VTIQ), or a combination thereof.

[0194] Certain embodiments relate to pharmaceutical formulations useful for reducing one or more of: mammographic breast density and breast firmness in a patient in need thereof, wherein the pharmaceutical formulation increases detection of breast cancer development in the patient.

[0195] Certain embodiments relate to pharmaceutical formulations useful for reducing one or more of: VBD %, AVBD, and breast firmness in a patient in need thereof, wherein the pharmaceutical formulation increases detection of breast cancer development in the patient.

[0196] Certain embodiments relate to pharmaceutical formulations useful for reducing one or more of: ABD %, AABD, and breast firmness in a patient in need thereof, wherein the pharmaceutical formulation increases detection of breast cancer development in the patient.

[0197] The pharmaceutical formulations disclosed herein can be used to affect one or more of the following in a patient: reducing mammographic breast density; treating mammographic breast density; reducing breast firmness; treating breast stiffness; reducing mammographic breast density in a patient with a breast having a mammographic breast density of 7.5% or greater; reducing Mammographic breast density in a patient with a breast score of 3 or 4 (or c or d); inducing breast involution in a patient; inducing net cell death exceeding proliferation in a patient's breast; inducing net extracellular matrix degradation exceeding extracellular matrix development in a patient's breast; a method of reversing breast cell number and mammographic breast density in a perimenopausal patient's breast; reducing mammographic breast density and perimenopausal symptoms in a patient, and preventing or treating autoimmune inflammatory mastitis (e.g., idiopathic inflammatory macromastia, plasma cell mastitis, granulomatous mastitis, and combinations of the foregoing).

[0198] These pharmaceutical formulations and combination treatments of androgens and aromatase inhibitors (Ai) as described herein can be used in premenopausal, perimenopausal, and / or postmenopausal women.

[0199] For example, it is known that high breast density in perimenopausal women is a risk for breast cancer development. The dense tissue of perimenopausal women is not considered normal and has pathological significance. This increase in breast density may be due to lifelong exposure to high levels of estrogen and progesterone in a low testosterone environment. The inventors have found that, among other factors, premenopausal, perimenopausal and / or postmenopausal women who receive an effective amount of androgen such as testosterone and an effective amount of aromatase inhibitor (such as anastrozole) through a multi-phase mode formulation can show a decrease in breast density and / or breast hardness. The inventors have also found that premenopausal, perimenopausal and / or postmenopausal women who receive an effective amount of testosterone and an effective amount of aromatase inhibitor (such as anastrozole) through a multi-phase mode formulation can show induction of breast involution and / or net cell death exceeding proliferation. The inventors have also found that an effective amount of aromatase inhibitor delivered to a subject by a multi-phase mode formulation given to the subject's breast tissue can be used to prevent the conversion of testosterone to estrogen, thereby allowing testosterone to cause breast cell involution.

[0200] Certain further embodiments are directed to preventing or treating autoimmune inflammatory mastitis (AIM), which includes the conditions of idiopathic inflammatory macromastitis, plasma cell mastitis, and granulomatous mastitis as described above.

[0201] Breast tissue may be a target tissue for autoimmune diseases, a process favored by the hormonal milieu (Touraine, 2005). Autoimmune breast tissue has been shown to have elevated aromatase (which converts androgens to estradiol) and other factors associated with inflammation, such as IGF2, EGFR, TGF-β, PDGFR-α and β, compared to normal breast tissue (Das, 2019).

[0202] The breast is unique in that it requires an immune-privileged lactation environment and rapid remodeling of tissues after lactation (termed lactational involution), which requires immunosuppression to avoid autoimmunity (Dawson, 2020). Central to this is the formation of regulatory T (Treg) cells, which are able to suppress the proliferation and cytokine production of effector T cells and play an important role in immune responses and the prevention of autoimmune diseases. During T cell development, T cell receptor (TCR) gene segments are rearranged to produce a diverse TCR repertoire necessary for immunity to invading pathogens. An unintended consequence of this diversity is the recognition of self-antigens, which can lead to autoimmunity. For T cells, before they are released into the periphery, two fundamental processes promote tolerance to self in the thymus (termed central tolerance): (1) negative selection, which eliminates autoreactive T cells; and (2) CD4 + Fork head frame P3 (Foxp3) +Generation of regulatory T (Treg) cells. Central to this process is the CD36-promoted transfer of cell surface antigens to promote tolerance to host antigens during homeostasis. The main regulator of Treg cell development and function is the transcription factor Foxp3. Testosterone treatment induces a strong increase in Treg cell populations both in vivo and in vitro (Walecki, 2015). Due to the immunosuppressive capacity of Treg cells, high levels of Treg cells in breast tumors reduce the ability of immunotherapy. Targeting Treg cells results in unacceptable autoimmune side effects, which limits the use of this approach. The effects of testosterone and aromatase inhibitors on breast tissue immune reactivity are shown in a graphical manner. Fig.17 .

[0203] The manifestation of AIM is associated with a dense stromal replacement of breast adipose tissue, which is clearly demonstrated on breast MRI in those affected by this disease (Touraine, 2005). One of the main players in this inflammatory cascade is CD36, a member of the class B scavenger receptor family of cell surface proteins. CD36 is present on platelets, erythrocytes, monocytes, differentiated adipocytes, skeletal muscle, mammary epithelial cells, splenocytes, and some skin microdermal endothelial cells. It has been demonstrated that a) CD36 is downregulated by estradiol in hormone-sensitive breast cancer cell lines (Uray, 2004); b) overexpression of CD36 in fibroblasts inhibits the formation of solid tumors in a subtype of breast cancer model (Cheng, 2020); and c) CD36 is severely repressed in non-malignant tissues of women at higher risk for the development of breast cancer due to high mammographic density (HMD) (i.e., breast tissue containing low fat) (DeFilippis, 2014).

[0204] Thea Tlsty's group (DeFilipis, 2014) proposed the following cascade: i) elevated basal DNA damage in HMD epithelial cells leads to increased activin A secretion; ii) activin A binds to its receptor on adjacent fibroblasts and activates the MAPK pathway; iii) MAPK pathway activation leads to PPARγ phosphorylation and inhibition; iv) PPARγ inhibition leads to decreased CD36 transcription and subsequent induction of the desmoplastic phenotype observed in HMD tissue.

[0205] As described herein, induction of CD36 expression in breast tissue can be obtained by treating the tissue with an effective amount of an androgenic agent in combination with an effective amount of an aromatase inhibitor, and the disclosure expressly extends to such methods and uses in patients such as perimenopausal, menopausal or postmenopausal women.

[0206] A subcutaneous pellet for preventing or treating an autoimmune disease or condition is described herein, which is used to promote a multi-phase sustained release mode to change the ratio of androgen to estrogen. As described herein, anastrozole and testosterone can be combined and pressed into pellets to provide the pharmacokinetics of this multi-phase sustained release of two active ingredients. This release pattern ensures that high levels of testosterone in the early stage are associated with very high levels of aromatase inhibitors, so only DHT is provided to the inflamed breast tissue, and testosterone is not converted into estradiol (which is pro-inflammatory). In the second stage, after reaching the peak testosterone level, the 5α-reductase induction from the first stage continues to ensure that high levels of DHT are maintained to exert an anti-inflammatory effect, while there is a faster decline in anastrozole. Due to the excessive decline of systemic estradiol, continued high levels of anastrozole may cause adverse events, but this risk will be reduced by the rapid reduction of anastrozole delivery. Then, the third delivery stage allows testosterone to be slowly released into the tissue, wherein 5α-reductase is induced in the somatic cell system to continue the disturbance of the antigenic environment. This unique multi-phase sustained release pattern can change tissue hormone levels without causing significant physiological changes in other serum reproductive hormone levels and the potential adverse side effects that result. The effects of the androgen testosterone and the aromatase inhibitor (anastrozole as described herein) on breast tissue immunoreactivity are shown in Fig.17 .

[0207] One of the advantages found in one or more of the disclosed compositions, delivery systems, and / or methods of use is the rapid early induction of the conversion of testosterone to dihydrotestosterone in tissues, wherein the early response can reset homeostatic mechanisms and reduce the ratio of estradiol to androgen in tissues in a beneficial manner. This may result in one or more of the following advantages:

[0208] A. Enhanced mammographic detection resulting from decreased breast density, enabling mammography to show malignancies at earlier and / or less aggressive stages.

[0209] B. Reduce the risk of interval breast cancers, such as those that may occur between rounds of mammography screening. These cancers are common in breasts with high MBD.

[0210] C. Reduce breast hardness.

[0211] D. Reduce pain during mammographic breast compression.

[0212] E. The ability to obtain better mammographic compression due, at least in part, to reduced pain.

[0213] F. Because better mammographic compression is achieved and breast tissue is less dense, less energy is required to expose the image on a mammogram, thereby reducing radiation exposure to breast tissue. Reduced risk of radiation-induced breast cancer.

[0214] G. The ability to achieve better patient compliance when undergoing regular mammography.

[0215] H. The ability to treat a patient without causing perturbations in the hypothalamic-pituitary axis and / or other endocrine axes.

[0216] I. Reduce breast pain in patients.

[0217] J. Reduce breast elasticity in patients.

[0218] K. Reducing mechanotransduction of the cellular genome to reduce the risk of malignant transformation in patients.

[0219] L. Increases the ratio of fibroglandular and adipose tissue in patients.

[0220] M. CD36 in patients.

[0221] N. Stabilizing and / or increasing the level of androgen receptor expression in a patient's breast tissue.

[0222] O. Treating macromastia in patients.

[0223] P. Increased GCDFP15 in patients.

[0224] Q. Reducing BPE in patient MRI images.

[0225] R. Reduce the size and / or number of cysts in patients.

[0226] S. Reduce the risk of breast cancer.

[0227] T. Treatment of autoimmune inflammatory mastitis (AIM) including conditions such as idiopathic inflammatory macromastitis, plasma cell mastitis, and granulomatous mastitis).

[0228] There are a variety of categories used by diagnosticians and physicians to characterize the type and / or degree of mammographic breast density of a patient's breasts.

[0229] The diagnosing or treating physician may use one or more examinations / tests to evaluate, characterize, and / or diagnose breast density, including, but not limited to, mammography, digital mammography, magnetic resonance imaging (MRI), ultrasound, digital breast tomosynthesis (DBT), visual tissue imaging quantitative (VTIQ), and combinations thereof. The physician may also use other indicators, such as medical or family history (to account for genetic predisposition to breast density) and / or qualitative assessment of MBD, such as (For example, the 5th edition uses the categories of breast parenchyma composition: “a” (breast is almost entirely fatty), “b” (scattered areas of fibroglandular density are present), “c” (breast is unevenly dense and may obscure small masses), and “d” (breast is very dense, reducing the sensitivity of mammography) (D'Orsi CJ, Sickles EA, Mendelson EB, Morris EA et al. (2013). ACR Atlas, Breast Impact Reporting and Data System. Reston, VA: American College of Radiology.

[0230] Breast pain is an important issue in women's health. Breast pain is also associated with an increase in VBD% and / or AVBD, breast stiffness, breast cancer risk, or a combination thereof. Certain embodiments relate to the use of androgen agents and aromatase inhibitors to reduce breast pain in patients.

[0231] Certain embodiments relate to the use of subcutaneous pellets comprising an effective amount of anastrozole and an effective amount of testosterone. In an exemplary embodiment, 0.5-10 mg of anastrozole (2,2'-[5-(1H-1,2,4-triazol-1-ylmethyl)-1,3-phenylene]bis(2-methylpropionitrile)) and 60-120 mg of testosterone in a pharmaceutical formulation compressed into a pellet are provided to the subject. In an exemplary embodiment, 4-6 mg of anastrozole and 60-120 mg of testosterone in a pharmaceutical formulation compressed into a pellet are provided to the subject. In an exemplary embodiment, about 4 mg of anastrozole and about 80 mg of testosterone in a pharmaceutical formulation compressed into a pellet are provided to the subject. Other androgenic agents may also be used. The duration of treatment for administering subcutaneous pellets comprising anastrozole and testosterone may vary between 2 weeks to 4 weeks, 3 months to 3 years, 6 months to 2 years, 3 months to 5 years, 1 year to 5 years, or 1 year to 3 years. In some embodiments, the duration of treatment may be about 2 weeks, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 2.5 years, or 3 years. In some embodiments, the duration of treatment may be at least 2 weeks, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 3 years, or 4 years. In some embodiments, the duration of treatment may be about 3 years. Treatment may be applied to one or more of the following: ABD% reduction, AABD reduction, VBD% reduction; AVBD reduction; breast pain reduction; breast hardness reduction; breast elasticity reduction; macromastia reduction; breast cyst reduction; mammography diagnosis sensitivity improvement and false positive reduction; fibroglandular and adipose tissue ratio increase; and androgen receptor expression level stabilization and / or increase.

[0232] Certain embodiments relate to the use of subcutaneous pellets comprising an effective amount of letrozole and an effective amount of testosterone. In an exemplary embodiment, a subcutaneous pellet comprising 0.5-20 mg of letrozole (4,4'-((1H-1,2,4-triazol-1yl)methylene)dibenzonitrile) and 40-130 mg of testosterone is provided to a patient for subcutaneous administration. In an exemplary embodiment, a subcutaneous pellet comprising approximately 10 mg of letrozole (4,4'-((1H-1,2,4-triazol-1yl)methylene)dibenzonitrile) and 40-130 mg of testosterone is provided to a patient for subcutaneous administration. Other androgenic agents may also be used. The duration of treatment for administering a subcutaneous pellet comprising letrozole and testosterone may vary between 2 weeks to 4 weeks, 3 months to 3 years, 6 months to 2 years, 3 months to 5 years, 1 year to 5 years, or 1 year to 3 years. In some embodiments, the duration of treatment may be about 2 weeks, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 2.5 years, or 3 years. In some embodiments, the duration of treatment may be at least 2 weeks, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 3 years, or 4 years. In some embodiments, the duration of treatment may be about 3 years. Treatment may be applied to one or more of the following: ABD% reduction, AABD reduction, VBD% reduction; AVBD reduction; breast pain reduction; breast hardness reduction; breast elasticity reduction; macromastia reduction; breast cyst reduction; mammography diagnosis sensitivity improvement and false positive reduction; fibroglandular and adipose tissue ratio increase; and androgen receptor expression level stabilization and / or increase.

[0233] Certain embodiments relate to the use of subcutaneous pellets comprising an effective amount of exemestane and an effective amount of testosterone. In an exemplary embodiment, a subcutaneous pellet comprising exemestane 10-75mg 6-methyleneandrosta-1,4-diene-3,17-dione and 40-130mg testosterone is provided to the patient for subcutaneous administration. Other androgen agents may also be used. The duration of treatment for administering exemestane and testosterone may vary between 2 weeks to 4 weeks, 3 months to 3 years, 6 months to 2 years, 3 months to 5 years, 1 year to 5 years or 1 year to 3 years. In certain embodiments, the duration of treatment may be approximately 2 weeks, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 2.5 years or 3 years. In certain embodiments, the duration of treatment may be at least 2 weeks, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 3 years or 4 years. In certain embodiments, the duration of treatment may be approximately 3 years. Treatment may be applied to one or more of the following: ABD% reduction, AABD reduction, VBD% reduction; AVBD reduction; breast pain reduction; breast firmness reduction; breast elasticity reduction; macromastia reduction; breast cyst reduction; improved sensitivity and reduced false positives in mammographic diagnosis; increased ratio between fibroglandular and adipose tissue; and stable and / or increased androgen receptor expression levels.

[0234] As described herein, the preparation of esters involves the functionalization of hydroxyl and / or carboxyl groups that may be present, as will be understood by those skilled in the art of pharmaceutical chemistry and drug delivery. For example, to prepare testosterone esters, the 17-hydroxyl of the testosterone molecule is typically reacted with a suitable organic acid under esterification conditions, such conditions typically involving the use of a strong acid, such as sulfuric acid, hydrochloric acid, etc., and a temperature sufficient to allow the reaction to proceed under reflux. If desired, conventional hydrogenolysis or hydrolysis procedures may be used to convert the ester back into a free acid.

[0235] The effective amount of androgen agent can be different because of androgen agent. In addition, the effective amount of testosterone may also be different every day. In some aspects, an effective amount of testosterone can be delivered in the form of a subcutaneous implant. In certain embodiments, the effective amount of testosterone can be 40 to 200 mg. For example, in certain embodiments, the effective amount of testosterone can be 40 to 120 mg, such as 20 mg, 40 mg, 60 mg, 80 mg, 100 mg or 120 mg.

[0236] In certain embodiments, an effective amount of testosterone can be delivered in the form of a subcutaneous implant, such as a subcutaneous pellet containing 40 to 200 mg of testosterone, such as 40 to 150 mg, 40 to 100 mg, 100 to 200 mg, 50 to 150 mg, 50 to 100 mg, 40 to 100 mg, 30 to 80 mg, 40 to 90 mg, 40 to 90 mg, 40 to 80 mg, 40 to 70 mg, 40 to 60 mg, 40 to 50 mg, 40 to 100 mg, 60 to 100 mg, 45 to 75 mg, or 40 to 45 mg of testosterone.

[0237] The effective amount of methyltestosterone may be different every day. In an exemplary embodiment, the effective amount of methyltestosterone can be 0.1mg to 10mg, such as 0.5mg to 9mg, 2mg to 8mg, 3mg to 7mg or 4mg to 5mg. For example, the effective amount of methyltestosterone can be 0.5mg, 1.25mg or 2.5mg.

[0238] The effective dose of androgen used in combination with aromatase inhibitors may be relatively lower than the standard dose because of low serum sex hormone-binding globulin levels in patients, which may be caused by the aromatase inhibitor.

[0239] Sex hormone binding globulin binds androgens (e.g., testosterone) and transports them throughout the body. Its production is regulated by a variety of mechanisms, but one of the factors affecting its level is the amount of estrogen in the serum: the higher the estrogen, the higher the sex hormone binding globulin, and the lower the free androgen. Conversely, the lower the estrogen, the lower the sex hormone binding globulin, and the higher the free androgen, which means that the androgen has a higher bioavailability. Therefore, after menopause, as estrogen levels decline, sex hormone binding globulin levels decline, and free androgens such as testosterone rise. This free androgen has multiple functions because androgen receptors are expressed in all cells of the body.

[0240] In certain embodiments, dosage levels below the lower limit of the above-mentioned androgenic range may be more than adequate, while in other cases, still larger dosages above the upper limit of the above-mentioned range may be employed without causing any deleterious side effects.

[0241] In certain embodiments, the aromatase inhibitor can be, for example, a steroidal aromatase inhibitor, a nonsteroidal aromatase inhibitor, and / or isomers thereof. The steroidal aromatase inhibitors developed to date are based on the basic androstenedione nucleus and incorporate chemical substituents at different positions on the steroid. Examples of steroidal aromatase inhibitors include, but are not limited to, exemestane. and formestane. Other examples include steroidal aromatase inhibitors based on mechanisms that mimic substrates that are converted by the enzyme into reactive intermediates and result in aromatase inactivation. In certain embodiments, the aromatase inhibitor is exemestane. Nonsteroidal aromatase inhibitors can be divided into three categories: aminoglutethimide-like molecules, imidazole / triazole derivatives, and flavonoid analogs. Examples of nonsteroidal aromatase inhibitors include anastrozole, exemestane, or letrozole. In certain embodiments, the aromatase inhibitor is anastrozole or letrozole. In certain embodiments, the aromatase inhibitor is anastrozole.

[0242] Aromatase inhibitors usually include third-generation aromatase inhibitors, such as anastrozole Exemestane and letrozole These third-generation aromatase inhibitors have transformed the treatment of women with hormone-sensitive breast cancer. The action of this class of aromatase inhibitors is specific because they actually eliminate estrogens from the serum, thereby reducing sex hormone-binding globulin, allowing for a synergistic effect.

[0243] In certain embodiments, the aromatase inhibitor may be selected from the group consisting of anastrozole, exemestane, or letrozole. In certain embodiments, the aromatase inhibitor is anastrozole or letrozole. In certain embodiments, the aromatase inhibitor is anastrozole.

[0244] In certain embodiments, a method for determining whether a patient has Breasts with a score of 3 or 4 (or c or d); breasts with a mammographic breast density of 7.5% or more; mammographically dense breasts; breasts with the same or more breast tissue as fat; breasts with more breast tissue than fat; methods of breast cancer, or a combination thereof.

[0245] In certain embodiments, the patient has or is diagnosed with A breast with a score (scale 1-4) in the range of 2 to 4, such as 2 to 3, or 3 to 4. In certain embodiments, the patient has or is diagnosed with Breasts with a score of 2 or more, such as Rated 3 or 4, or Rated 4.

[0246] In certain embodiments, the patient has or is diagnosed with A breast with a score (grade ad) in the range of b to d, such as b to c, or c to d. In certain embodiments, the patient has or is diagnosed with Breasts with a score of B or higher, such as A grade of C or D, or Rated d.

[0247] In certain embodiments, the patient has or is diagnosed with a breast having a mammographic breast density of 7.5% or greater, such as a mammographic breast density of 10% or greater, 15% or greater, 20% or greater, 30% or greater, 50% or greater, 70% or greater, or 95% or greater.

[0248] In certain embodiments, the patient has or is diagnosed with a breast having a VBD% of 7.5% or more, e.g., a VBD% of 10% or more, 15% or more, 20% or more, 30% or more, 50% or more, 70% or more, or 95% or more.

[0249] In certain embodiments, the patient has or is diagnosed with a breast having an ABD% of 7.5% or more, e.g., an ABD% of 10% or more, 15% or more, 20% or more, 30% or more, 50% or more, 70% or more, or 95% or more.

[0250] In certain embodiments, the patient has or is diagnosed with A breast with a score of 3 (or C) and a mammographic breast density of 7.5% or more, such as a mammographic breast density of 10% or more, 15% or more, 20% or more, 30% or more, 50% or more, 70% or more, or 95% or more. In certain embodiments, the patient has or is diagnosed with Breasts with a score of 4 (or D) and a mammographic breast density of 7.5% or greater, e.g., a mammographic breast density of 10% or greater, 15% or greater, 20% or greater, 30% or greater, 50% or greater, or 95% or greater.

[0251] In certain embodiments, the patient has or is diagnosed with a breast having a VBD% in the range of 1% to 100%, such as a VBD% of 1% to 24%, 5% to 100%, 5% to 95%, 5% to 90%, 5% to 80%, 5% to 70%, 5% to 60%, 5% to 50%, 5% to 40%, 5% to 30%, 5% to 25%, 5% to 20%, 10% to 100%, 10% to 95%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 25%, 10% to 20%, 25% to 100%, 25% to 75%, or 25% to 100%. or a VBD % of 75% to 90%. In certain embodiments, the patient has or is diagnosed as having a breast with a VBD% in the range of 10% to 40%.

[0252] In certain embodiments, the patient has or is diagnosed with a breast with an ABD% in the range of 1% to 100%, such as an ABD% of 1% to 24%, 5% to 100%, 5% to 95%, 5% to 90%, 5% to 80%, 5% to 70%, 5% to 60%, 5% to 50%, 5% to 40%, 5% to 30%, 5% to 25%, 5% to 20%, 10% to 100%, 10% to 95%, 10% to 90%, 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 25%, 10% to 20%, 25% to 100%, 25% to 75%, or 25% to 100%. or an ABD % of 75% to 90%. In certain embodiments, the patient has or is diagnosed with a breast having an ABD% in the range of 50% to 100%. In certain embodiments, the patient has or is diagnosed with a mammographically dense breast, such as a breast having approximately the same or more breast tissue as fat.

[0253] In certain embodiments, the patient is a perimenopausal woman or a postmenopausal woman. In certain embodiments, the patient is a perimenopausal woman.

[0254] In certain embodiments, the pharmaceutical formulation can be used to reduce or decrease the patient's need for mammography between one or more annual interventional mammography examinations. For example, a drug agent could be used to keep a patient on track between one or more annual interventional mammograms. The score is reduced or decreased by 1 point or more, such as by 2 points or more, 3 or 4 points or 4 points between one or more annual interventional mammography examinations. In certain embodiments, the pharmaceutical formulation can be used to reduce the patient's score between one or more annual interventional mammography examinations. The score is reduced or decreased by 1 point, such as by 2, 3, or 4 points between one or more annual interventional mammography examinations. In certain embodiments, the drug formulation maintains or stabilizes the patient's score between one or more annual interventional mammography examinations. score.

[0255] The time period for one or more annual interventional mammography examinations can be 1 to 20 years, such as 1 year, 1.5 years, 2 years, 3 years, 4 years, 5 years, 6 years, 10 years, 15 years, or 20 years. The time period for one or more annual interventional mammography examinations can be 1 year, 2 years, 4 years, 5 years, 7 years, 10 years, 15 years, or 20 years.

[0256] In certain embodiments, the pharmaceutical formulations may be used to reduce or decrease the mammographic breast density of a patient's breast between one or more annual interventional mammographic examinations. In certain embodiments, the pharmaceutical formulations may be used to reduce or decrease the VBD% and / or AVBD of a patient's breast between one or more annual interventional mammographic examinations. In certain embodiments, the pharmaceutical formulations may be used to reduce or decrease the ABD% and / or AABD of a patient's breast between one or more annual interventional mammographic examinations. For example, the pharmaceutical formulation can be used to reduce or decrease the mammographic breast density of a patient's breast between one or more annual interventional mammographic examinations by a range of 1% to 99%, such as a range of 1% to 80%, 1% to 50%, 1% to 30%, 1% to 20%, 1% to 10%, 3% to 40%, 3% to 20%, 5% to 60%, 5% to 25%, 5% to 15%, 5% to 10%, 10% to 60%, 10% to 40%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 60%, 20% to 40%, 20% to 30%, 30% to 60%, 30% to 50%, or 30% to 40% between one or more annual interventional mammographic examinations. For example, the pharmaceutical formulation can be used to reduce or decrease the VBD% and / or AVBD of a patient's breast between one or more annual interventional mammographic examinations in a range of 1% to 99%, such as a reduction or decrease of 1% to 80%, 1% to 50%, 1% to 30%, 1% to 20%, 1% to 10%, 3% to 40%, 3% to 20%, 5% to 60%, 5% to 25%, 5% to 15%, 5% to 10%, 10% to 60%, 10% to 40%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 60%, 20% to 40%, 20% to 30%, 30% to 60%, 30% to 50%, or 30% to 40% between one or more annual interventional mammographic examinations. For example, the pharmaceutical formulation can be used to lower or reduce the ABD% and / or AABD of a patient's breast between one or more annual interventional mammographic examinations by a range of 1% to 99%, such as a range of 1% to 80%, 1% to 50%, 1% to 30%, 1% to 20%, 1% to 10%, 3% to 40%, 3% to 20%, 5% to 60%, 5% to 25%, 5% to 15%, 5% to 10%, 10% to 60%, 10% to 40%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 60%, 20% to 40%, 20% to 30%, 30% to 60%, 30% to 50%, or 30% to 40% between one or more annual interventional mammographic examinations.For example, the pharmaceutical formulation can be used to reduce or decrease the mammographic breast density of a patient's breast by at least 2% between one or more annual interventional mammographic examinations, for example, by at least 5%, 10%, 20%, 30%, 40%, 50%, 75%, 85%, 95%, or 99% between one or more annual interventional mammographic examinations. For example, the pharmaceutical formulation can be used to reduce or decrease the VBD% and / or AVBD of a patient's breast by at least 2% between one or more annual interventional mammographic examinations, for example, by at least 5%, 10%, 20%, 30%, 40%, 50%, 75%, 85%, 95%, or 99% between one or more annual interventional mammographic examinations. For example, the pharmaceutical formulation can be used to reduce or decrease the ABD% and / or AABD of a patient's breast by at least 2% between one or more annual interventional mammographic examinations, such as by at least 5%, 10%, 20%, 30%, 40%, 50%, 75%, 85%, 95%, or 99% between one or more annual interventional mammographic examinations. In certain embodiments, the pharmaceutical formulation can be used to maintain or stabilize the mammographic breast density of a patient's breast between one or more annual interventional mammographic examinations.

[0257] In certain embodiments, the pharmaceutical formulation can be used to reduce or decrease the mammographic breast density of a patient's breast by at least 2%, e.g., 5%, 10%, 20%, or 30% over a 4 hour period, e.g., over a 8 hour, 24 hour, 1 day, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or 1 year period.

[0258] In certain embodiments, the pharmaceutical formulation can be used to reduce or decrease the VBD% and / or AVBD of a patient's breast by at least 2%, e.g., 5%, 10%, 20%, or 30% over a 4 hour period, e.g., over a 8 hour, 24 hour, 1 day, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or 1 year period.

[0259] In certain embodiments, the pharmaceutical formulation can be used to reduce or decrease the ABD% and / or AABD of a patient's breast by at least 2%, e.g., 5%, 10%, 20%, or 30% over a 4 hour period, e.g., over a 8 hour, 24 hour, 1 day, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or 1 year period.

[0260] In certain embodiments, the pharmaceutical preparation can be used to reduce or reduce the risk of breast cancer development in a patient. For example, in certain embodiments, the pharmaceutical preparation can be used to reduce or reduce the risk of breast cancer development in a patient between one or more annual interventional mammography tests. In certain embodiments, the pharmaceutical preparation can be used to reduce or reduce the risk of breast cancer development in a patient between one or more annual interventional mammography tests, and avoid, reduce, reduce or reverse one or more perimenopausal symptoms. For example, one or more perimenopausal symptoms that can be reduced, reduced or avoided may include, but are not limited to, irregular menstruation; hot flashes and sleep problems; mood changes; mood swings; irritability; depression; vaginal dryness; urinary tract infection or vaginal infection; urinary incontinence; reduced fertility; changes in sexual arousal or demand; bone loss; bone fragility; osteoporosis; or changes in cholesterol levels, such as increased low-density lipoprotein (LDL) cholesterol, decreased high-density lipoprotein (HDL) cholesterol; or a combination thereof.

[0261] In certain embodiments, the pharmaceutical formulation can be used to increase or improve a patient's fat to breast tissue ratio between one or more annual interventional mammography examinations. For example, the pharmaceutical formulation increases or improves a patient's fat to breast tissue ratio from 1:19 to 19:1 between one or more annual interventional mammography examinations, for example, increases or improves a patient's fat to breast tissue ratio from 1:15 to 19:1, from 1:10 to 19:1, from 1:5 to 19:1, from 1:2 to 19:1, from 2:3 to 19:1, between one or more annual interventional mammography examinations. from 2:1 to 19:1, from 4:1 to 19:1, from 6:1 to 19:1, from 8:1 to 19:1, from 10:1 to 19:1, from 1:19 to 10:1, from 1:10 to 10:1, from 1:4 to 10:1, from 1:2 to 10:1, from 3:2 to 10:1, from 3:1 to 10:1 Increase or improve from 5:1 to 10:1, increase or improve from 7:1 to 10:1, increase or improve from 9:1 to 10:1, increase or improve from 15:1 to 10:1, increase or improve from 1:15 to 5:1, increase or improve from 1:5 to 5:1, increase or improve from 1:3 to 5:1, increase or improve from 3:2 to 5:1, increase or improve from 3:1 to 5:1, increase or improve from 6:1 to 5:1, increase or improve from 8:1 to 5:1, from 10:1 to 5:1, from 1:19 to 3:1, from 1:10 to 3:1, from 1:4 to 3:1, from 1:2 to 3:1, from 2:1 to 3:1, from 4:1 to 3:1, from 6:1 to 3:1, from 8:1 to 3:1, from 10:1 to 3:1, or from 15:1 to 3:1.

[0262] In certain embodiments, the pharmaceutical formulation can be used to increase or improve a patient's fat to breast tissue ratio from 1:19 to 19:1, e.g., from 1:10 to 19:1, from 1:5 to 19:1, from 1:2 to 19:1, from 2:3 to 19:1, from 2:1 to 19:1, over a 4 hour period, over an 8 hour period, over a 24 hour period, over a 3 day period, over a 1 week period, over a 2 week period, over a 1 month period, over a 2 month period, over a 3 month period, over a 6 month period, over a 9 month period, over a 1 year period, or over a 5 year period.

[0263] In certain embodiments, the pharmaceutical formulation may be used to increase the percentage of fat in a patient's breast between one or more annual interventional mammograms. For example, the pharmaceutical formulation increases the percentage of fat in the breasts of treated patients between one or more annual interventional mammographic examinations by 1% to 99%, such as between one or more annual interventional mammographic examinations by 1% to 90%, 1% to 70%, 1% to 50%, 1% to 30%, 1% to 20%, 1% to 15%, 1% to 10%, 3% to 60%, 3% to 20%, 5% to 70%, 5% to 50%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 60%, 10% to 40%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 50%, 20% to 30%, 30% to 60%, 30% to 50%, or 30% to 40% between one or more annual interventional mammographic examinations.

[0264] In certain embodiments, the pharmaceutical formulation increases the percentage of fat in the breast of a treated patient by at least 2%, e.g., at least 5%, at least 10%, at least 25%, at least 40%, at least 75%, at least 95%, or at least 99% over a 4 hour period, over an 8 hour period, over a 24 hour period, over a 3 day period, over a 1 week period, over a 2 week period, over a 1 month period, over a 2 month period, over a 3 month period, over a 6 month period, over a 9 month period, over a 1 year period, or over a 5 year period.

[0265] In certain embodiments, the pharmaceutical formulation enhances, increases, or improves mammographic visualization of the breast or breast compression during the examination between one or more annual interventional mammographic examinations. For example, the pharmaceutical formulation enhances, increases, or improves mammographic visualization of the breast or breast compression during the examination between one or more annual interventional mammographic examinations by 5% to 70%, 5% to 50%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 50%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 60%, 20% to 40%, 20% to 30%, 30% to 70%, 30% to 50%, or 30% to 40%. In certain embodiments, as a result of enhancing, increasing, or improving mammographic visualization of the breast or breast compression during the examination, the pharmaceutical formulation further alleviates or reduces patient pain during breast compression. For example, the pharmaceutical formulation further reduces, decreases or minimizes patient pain during breast compression in the range of 5% to 80%, 5% to 50%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 80%, 10% to 60%, 10% to 40%, 10% to 20%, 10% to 15%, 20% to 70%, 20% to 50%, 20% to 30%, 30% to 70%, 30% to 50%, or 30% to 40% less pain as a result of enhanced, increased or improved breast compression during mammographic visualization or detection of the breast.

[0266] In certain embodiments, the pharmaceutical formulation alleviates or reduces patient pain during breast compression. For example, the pharmaceutical formulation alleviates or reduces patient pain during breast compression between one or more annual interventional mammographic examinations by 5% to 80%, 5% to 60%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 80%, 10% to 60%, 10% to 40%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 70%, 20% to 50%, 20% to 30%, 30% to 90%, 30% to 50%, or 30% to 40%. In certain embodiments, as a result of alleviating, reducing or minimizing patient pain during breast compression, the pharmaceutical formulation further enhances, increases or improves breast compression during mammographic visualization or examination of the breast between one or more annual interventional mammographic examinations. For example, the pharmaceutical agent further enhances, increases or improves mammographic visualization of the breast or breast compression during examinations between one or more annual interventional mammographic examinations by 5% to 80%, 5% to 60%, 5% to 40%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 80%, 10% to 60%, 10% to 40%, 10% to 20%, 10% to 15%, 20% to 80%, 20% to 60%, 20% to 30%, 30% to 80%, 30% to 50%, or 30% to 40%.

[0267] In certain embodiments, the pharmaceutical formulation alleviates or reduces patient pain based on a visual analog scale (VAS) during breast compression. For example, the pharmaceutical formulation alleviates or reduces patient pain based on a VAS during breast compression such that the patient does not experience significant pain of 50-100 mm, 50-80 mm, 50-70 mm, 60-100 mm, 70-100 mm, 80-100 mm, or 90-100 mm during one or more mammographic examinations or during one or more annual interventional mammographic examinations.

[0268] In certain embodiments, the pharmaceutical formulation enhances or improves patient compliance with periodic mammographic visualization or testing, such as compliance with mammographic visualization or testing every 6 months, yearly, every 2 years, every 3 years, or every 5 years.

[0269] In certain embodiments, the pharmaceutical formulation mitigates or reduces the amount of radiation exposure required to visualize or detect a patient's breast during one or more subsequent mammograms, such as during one or more subsequent annual mammograms. For example, the pharmaceutical formulation mitigates or reduces the amount of radiation exposure required to visualize or detect a patient's breast during one or more subsequent mammograms, such as during one or more subsequent annual mammograms by 5% to 99%, 5% to 80%, 5% to 70%, 5% to 50%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 80%, 10% to 60%, 10% to 40%, 10% to 20%, 10% to 15%, 20% to 80%, 20% to 60%, 20% to 40%, 20% to 30%, 30% to 80%, 30% to 60%, 30% to 50%, or 30% to 40%.

[0270] In certain embodiments, the pharmaceutical formulation induces breast involution in a patient's breast, such as a perimenopausal patient's breast.

[0271] In certain embodiments, the pharmaceutical formulation induces breast cell involution in a patient's breast, such as a perimenopausal patient's breast.

[0272] In certain embodiments, the pharmaceutical formulation induces net cell death exceeding proliferation in a patient's breast, such as a perimenopausal patient's breast.

[0273] In certain embodiments, the pharmaceutical formulation reverses cellularity and mammographic breast density in a patient's breast, such as a perimenopausal patient's breast.

[0274] In certain embodiments, the pharmaceutical preparation alleviates or reduces the breast hardness in the breast of a patient, such as a perimenopausal patient. For example, the pharmaceutical preparation alleviates or reduces the breast hardness in the breast of a patient between one or more annual interventional mammography tests by 5% to 80%, 5% to 60%, 5% to 40%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 80%, 10% to 60%, 10% to 40%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 80%, 20% to 60%, 20% to 40%, 20% to 30%, 30% to 80%, 30% to 60%, or 30% to 40%. For example, the pharmaceutical preparation alleviates or reduces the breast hardness in the patient's breast by at least 5%, such as at least 8%, at least 10%, at least 15%, at least 20% or at least 30% each year. In certain embodiments, the pharmaceutical formulation reduces or decreases breast firmness in a patient's breast by at least 5%, e.g., at least 8%, at least 10%, at least 15%, at least 20%, or at least 30%, over a 4 hour period, e.g., over an 8 hour period, a 24 hour period, a 3 day period, a 1 week period, a 2 week period, a 1 month period, a 2 month period, a 3 month period, a 6 month period, a 9 month period, a 1 year period, or a 5 year period.

[0275] In certain embodiments, the pharmaceutical formulation enhances, increases or improves mammographic visualization or detection of a patient's breast, such as a perimenopausal patient's breast. For example, the pharmaceutical formulation enhances, increases or improves mammographic visualization or detection of a patient's breast between one or more annual interventional mammographic examinations by 5% to 80%, 5% to 50%, 5% to 30%, 5% to 20%, 5% to 15%, 5% to 10%, 10% to 80%, 10% to 60%, 10% to 30%, 10% to 20%, 10% to 15%, 20% to 80%, 20% to 60%, 20% to 30%, 30% to 80%, 30% to 60%, or 30% to 40%. In certain embodiments, the pharmaceutical formulation enhances, increases or improves mammographic visualization or detection of a patient's breast by at least 5%, e.g., at least 10%, at least 15%, at least 25%, at least 40%, at least 50%, or at least 75%, over a 4 hour period or over other time periods, e.g., 8 hours, 24 hours, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, 1 year, or 5 years.

[0276] In certain embodiments, the pharmaceutical formulations reduce mammographic breast density and avoid inducing masculinizing androgenic side effects or inducing a hyperandrogenic state. For example, masculinizing androgenic side effects may include male pattern baldness, hirsutism or increased hair growth in areas that the patient does not want, deepening of the voice, acne, or a combination thereof. In certain embodiments, the pharmaceutical formulations reduce mammographic breast density and avoid inducing masculinizing androgenic side effects or inducing a hyperandrogenic state. In certain embodiments, the pharmaceutical formulations reduce mammographic breast density and minimize the induction of masculinizing androgenic side effects or the induction of a hyperandrogenic state.

[0277] In certain embodiments, the pharmaceutical preparation significantly improves or improves the patient's physical function, such as physical function related to the patient's central nervous system, libido, musculoskeletal system, cardiovascular system, risk of infection with autoimmune diseases, severity of symptoms associated with autoimmune diseases, or a combination thereof. For example, when associated with the patient's central nervous system, the pharmaceutical preparation can reduce depression, anxiety, general cognitive dysfunction including memory, or reduce the risk of dementia and Parkinson's disease. For example, when associated with the patient's libido, the pharmaceutical preparation can significantly improve overall libido, including sexual arousal speed and the ability to reach orgasm. For example, when associated with the patient's musculoskeletal system, the pharmaceutical preparation can reduce inflammation and degenerative arthritis, improve bone mineral density, or improve muscle strength. For example, when associated with the patient's cardiovascular system, the pharmaceutical preparation can reduce foamy macrophage deposition in the arterial wall, reduce atherosclerosis, increase high-density lipoprotein, leading to improved cholesterol, or high-density lipoprotein ratio. For example, when associated with the risk of infection with an autoimmune disease in a patient, the pharmaceutical preparation can significantly reduce or reduce the risk of infection with an autoimmune disease in a treated patient, such as Sjögren's syndrome, lupus, and rheumatoid arthritis. For example, the pharmaceutical formulation can significantly reduce or reduce the severity of symptoms associated with treating an autoimmune disease in a patient, such as Sjögren's syndrome, lupus, and rheumatoid arthritis, when related to the severity of symptoms associated with the patient's autoimmune disease. In certain embodiments, the pharmaceutical formulation significantly improves or improves a patient's physical function, such as cognitive function; reduces degenerative CNS diseases, including dementia or Parkinson's disease; muscle strength; libido; vitality; reduces anxiety and depression caused by monoamine oxidase; or a combination thereof.

[0278] In certain embodiments, the pharmaceutical formulation further provides one or more of the following: i) reduction in mammographic breast density; ii) increased involution of the patient's breast without converting testosterone to estrogen; iii) significantly reduced, reduced, or reversed perimenopausal symptoms; or iv) significantly improved or improved physical function in the patient, including cognitive function; reduced symptoms associated with degenerative CNS diseases, including dementia or Parkinson's disease; muscle strength; libido; vitality; reduced anxiety and depression caused by monoamine oxidase; or a combination thereof. In certain embodiments, the pharmaceutical formulation further provides one or more of the following: i) reduction in mammographic breast density; ii) increased involution of hormonally affected end organs, including the breast, without converting testosterone to estrogen; iii) significantly reduced, reduced, or reversed perimenopausal symptoms associated with fluctuations in estrogen levels; or iv) significantly improved or improved physical function in the patient, including cognitive function; reduced symptoms associated with degenerative CNS diseases, including dementia or Parkinson's disease; muscle strength; libido; vitality; reduced anxiety and depression caused by monoamine oxidase; or a combination thereof.

[0279] In certain embodiments, the patient has a high free androgen index level in the breast, e.g., 30% or more, within four hours of administering an androgen and an aromatase inhibitor. In certain embodiments, the patient has a supraphysiological free androgen index level in the breast within four hours of administering an androgen and an aromatase inhibitor.

[0280] In certain embodiments, treatment with a pharmaceutical formulation or combination therapy of an androgen agent and an aromatase inhibitor as described herein further comprises: a) measuring free androgen index levels and / or aromatase inhibitor levels in serum isolated from a blood sample taken from the patient at least one month after treatment; b) determining a subsequent dose comprising a subsequent effective amount of the androgen agent and a subsequent effective amount of the aromatase inhibitor; and c) administering the subsequent dose to the patient.

[0281] In certain embodiments, treatment with a pharmaceutical formulation or combination therapy of an androgen agent and an aromatase inhibitor as described herein further comprises: a) measuring the free androgen index level and / or the aromatase inhibitor level in serum separated from a blood sample taken from the patient at least 1 month after treatment, comprising centrifuging the patient's blood sample to separate the serum; b) determining a subsequent dose, comprising a subsequent effective amount of the androgen agent and a subsequent effective amount of the aromatase inhibitor; and c) administering the subsequent dose to the patient.

[0282] In certain embodiments, the free androgen index serum level measured in treated patients after 1 month may be 10-25%, such as 10-20%, 10-15%, 15-25%, 15-20%, 12-18%, 8-15% or 11-14%.

[0283] In certain embodiments, the free androgen index serum level measured in treated patients after 3 months may be 2-10%, such as 2-8%, 2-6%, 2-5%, 2-4%, 4-10%, 5-8%, 3-7%, 4-6%, 3-6%, 4-7%, 5-10% or 2-5%.

[0284] In certain embodiments, the administration of an aromatase inhibitor reduces the aromatization of testosterone to estrogen in the subcutaneous fat of the treated patient, for example, by reducing aromatization by 80-95% or 100%. For example, the administered aromatase inhibitor can reduce the aromatization of testosterone to estrogen in the patient's breast subcutaneous fat, the patient's pelvic subcutaneous fat, the patient's buttocks subcutaneous fat, the patient's abdominal subcutaneous fat, or a combination thereof, for example, by reducing aromatization by 80-95% or 100%.

[0285] In certain embodiments, the administration of an aromatase inhibitor reduces the aromatization of adrenal androgens such as androstenedione to estrogens in the subcutaneous fat of the treated patient, for example, reducing aromatization by 80-95% or 100%. For example, the aromatase inhibitor administered can reduce the aromatization of testosterone to estrogens in the patient's breast subcutaneous fat, the patient's pelvic subcutaneous fat, the patient's buttocks subcutaneous fat, the patient's abdominal subcutaneous fat, or a combination thereof, for example, reducing aromatization by 80-95% or 100%.

[0286] Annual mammographic density screening can be performed to determine reduction in breast density using an appropriate mammographic algorithm that measures the volume of fibroglandular tissue as a percentage of total breast volume (MBD). When this is a function of the average of the two breast densities, the goal is to achieve a MBD of less than 10%. The rate of reduction in breast density should be at least 2% per year. If 2% is not achieved in the first year, a 10% annual boost factor can be introduced in TD and AD (V1(N) above). This annual boost factor is generally only introduced on an annual basis if the increase in androgen index (AI) is less than 10%.

[0287] Annual mammographic density screening can be performed to determine reduction in breast density using an appropriate mammographic algorithm that measures the volume of fibroglandular tissue as a percentage of total breast volume (VBD%). When this is a function of the average of the two breast densities, the goal is to achieve a VBD% of less than 10%. The rate of reduction in breast density (VBD%) should be at least 2% per year. If 2% is not achieved in the first year, a 10% annual boost factor can be introduced in TD and AD (V1(N) above). This annual boost factor is generally only introduced on an annual basis if the increase in androgen index (AI) is less than 10%.

[0288] In certain embodiments, the use of the combination of an aromatase inhibitor and testosterone results in an increase in the bioavailability of dihydrotestosterone by 25% to 75%, 35% to 65%, or 45% to 55%. In certain embodiments, the increase in the bioavailability of dihydrotestosterone is greater than 25%, greater than 35%, greater than 45%, or greater than 55%.

[0289] For example, the androgen agent for the pharmaceutical preparation or combination therapy as described herein can be selected from the group consisting of testosterone, methyltestosterone and / or dehydroepiandrosterone. In certain embodiments, the androgen agent can be testosterone undecanoate, such as about 40 mg testosterone undecanoate. For example, the aromatase inhibitor can be selected from the group consisting of anastrozole, exemestane or letrozole. In some aspects, the aromatase inhibitor can be anastrozole, such as about 1 mg of anastrozole.

[0290] In certain embodiments, the pharmaceutical formulation may include administering a pharmaceutical formulation comprising an androgen or androgen / aromatase inhibitor complex linked to an aromatase inhibitor, for example, by an ester bond, wherein the complex is produced by methods known in the art.

[0291] In preferred embodiments, the androgenic agent (eg, testosterone, methyltestosterone and / or dehydroepiandrosterone) and the aromatase inhibitor (eg, anastrozole, exemestane or letrozole) are both administered subcutaneously, for example as an implant such as a pellet.

[0292] In certain embodiments, the androgen (e.g., testosterone, methyltestosterone and / or dehydroepiandrosterone) and aromatase inhibitor (e.g., anastrozole, exemestane or letrozole) are administered subcutaneously, e.g., as a pellet. For example, testosterone and anastrozole can be administered subcutaneously.

[0293] The appropriate dosing regimen using androgens, aromatase inhibitors, or pharmaceutical preparations comprising androgens and aromatase inhibitors, the amount of each dose administered, and the intervals between compound doses may depend on various factors, such as the specific aromatase inhibitor and androgen used in combination, the type of pharmaceutical preparation used, the type of physiological condition treated, the characteristics of the subject treated (e.g., species, age, weight, sex, medical condition, feeding / fasting), route of administration, and the severity of the condition treated, or a combination thereof. A physician or diagnostician with ordinary skills can easily determine and prescribe an effective amount of androgens, aromatase inhibitors, or pharmaceutical preparations to prevent or treat a specific physiological condition.

[0294] The pharmaceutical preparation or formulation to be administered may contain an amount of the compound or a pharmaceutically acceptable salt or ester thereof, an amount effective to treat the condition of the subject being treated. Because two different compounds may be used together in a combination therapy, the potency of each compound and the interactive effects achieved by combining them together will also generally be considered. Consideration of these factors for the purpose of determining a therapeutically effective or prophylactically effective dose required to ameliorate side effects is well within the scope of the ordinary skilled clinician.

[0295] Administration of the androgen agent and aromatase inhibitor or a pharmaceutical formulation comprising the combination thereof to a subject includes self-administration and administration to the subject by another person (eg, a physician, nurse, health care worker, friend, etc.).

[0296] In certain embodiments, the pharmaceutical preparation can be formulated in a manner compatible with the desired results. The pharmaceutical preparation can be administered in a convenient formulation. The following formulation examples are illustrative only and are not intended to limit the scope of the present disclosure.

[0297] In certain embodiments, the pharmaceutical preparation can be an implant or pellet prepared by direct compression. For example, a pellet preparation can incorporate a diluent, a binder, a lubricant and a disintegrant as well as an active ingredient. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or calcium sulfate, inorganic salts such as sodium chloride and powdered sugar. Powdered cellulose derivatives are also useful. Natural and synthetic gums are also convenient, including gum arabic, alginate, methylcellulose and / or polyvinyl pyrrolidine. Polyethylene glycol, ethyl cellulose and wax can also be used as adhesives. A pellet preparation may require a lubricant to prevent the pellet and punch from sticking in the mold. The lubricant can be selected from a smooth solid such as talc, magnesium stearate and calcium stearate, stearic acid and / or hydrogenated vegetable oil. Typically, the implant or pellet hardness range of the implant or pellet is about 6Kg / N to about 10Kg / N, preferably about 7Kg / N to about 9Kg / N, most preferably about 8Kg / N, and all such dosage forms within this range are clearly provided herein.

[0298] By compressing the formulation into a solid dosage form, such as an implant in the form of a pellet for subcutaneous administration, a sustained release multiphasic concentration pattern as described herein can be obtained. After knowing that such release patterns of androgens and aromatase inhibitors can be obtained, in view of the disclosure provided herein, one of ordinary skill in the art will be able to use different ingredients in the formulation and provide such pharmaceutical formulations according to the present disclosure.

[0299] In certain embodiments, the implant or pellet can be inserted into the subcutaneous fat of the subject's pelvis, the subcutaneous fat of the subject's breasts, the subcutaneous fat of the subject's buttocks, the subcutaneous fat of the subject's abdomen, or a combination thereof. In certain embodiments, the implant or pellet can be inserted into the subcutaneous fat of the subject's lower abdominal wall. In certain embodiments, the implant or pellet can be inserted into the subcutaneous fat of the subject's upper gluteal region.

[0300] As will be appreciated, the androgen agent and the aromatase inhibitor may be provided in a pharmaceutical formulation other than for subcutaneous administration, either together in the same formulation or in separate formulations for use in combination therapy as described herein, such formulations comprising one or more suitable fillers, lubricants, binders, disintegrants and / or pharmaceutically acceptable carriers or excipients as described above, or for providing a pharmaceutical formulation for a desired route of administration and / or for obtaining a desired release profile (e.g., a sustained release multiphasic release profile) of one or more actives.

[0301] In certain embodiments, androgen, aromatase inhibitor or pharmaceutical preparation comprising a combination thereof can be provided in the form of a product, such as a kit, which includes an active ingredient therein or an active ingredient in a suitable pharmaceutical preparation, packaged for distribution. The kit may also include instructions for using the kit components in one or more disclosed methods. The instructions may include instructions for practicing one or more disclosed methods. Thus, for example, the kit may include an androgen or aromatase inhibitor in a pharmaceutical preparation in a container, package or dispenser and instructions for application to a human subject. The instructions may also include a satisfactory clinical endpoint or an indication of any adverse symptoms that may occur, or any other information required by the Food and Drug Administration for use in humans.

[0302] The instructions may be on "printed matter", e.g., on paper or cardboard within the kit, or on a label affixed to the kit or packaging material, or affixed to a vial or tube containing the kit components. The instructions may also be contained on a computer readable medium, such as a disk (floppy or hard disk), an optical CD (e.g., CD or DVD-ROM / RAM), a magnetic tape, an electronic storage medium such as RAM and ROM), and a mixture of these including magnetic / optical storage media.

[0303] Although the present disclosure has been described according to certain exemplary embodiments in order to facilitate a better understanding of the present disclosure, it should be understood that various modifications can be made without departing from the principles disclosed herein. Therefore, the present invention should be understood to include such modifications within its scope.

[0304] Examples

[0305] Example 1: Single-dose trial to evaluate the pharmacokinetics of testosterone and anastrozole from subcutaneous testosterone and anastrozole implants (80 mg / 4 mg (T+Ai)) in women

[0306] Pharmacokinetic analysis was performed on subcutaneous testosterone 80 g and anastrozole 4 mg implants in 11 healthy volunteers with high mammographic breast density. The study evaluated the pattern of both anastrozole and testosterone and allowed modeling of the pharmacokinetics of this exemplary implant to demonstrate the pattern and end-organ pharmacodynamic response known to exist in excess aromatase activity, i.e., in the tissues of women with high mammographic breast density. This was evaluated by directly measuring breast tissue elasticity in relation to mammographic breast density.

[0307] Ultrasonic evaluation of pellet dissolution was performed to demonstrate how a comprehensive dissolution analysis of reproductive hormone infusions can be performed to evaluate potential effects on hypothalamic-pituitary function.

[0308] Main objectives:

[0309] The pharmacokinetic characteristics of testosterone after a single dose of T+Ai were evaluated by subcutaneous implantation.

[0310] The pharmacokinetic characteristics of anastrozole after a single dose of T+Ai were evaluated by subcutaneous implantation.

[0311] Secondary Objectives

[0312] To investigate the potential effects of shear wave ultrasound assessment on breast tissue elasticity.

[0313] Implant preparation: Testosterone-Anastrozole Pellets

[0314] Pellets for subcutaneous insertion are prepared by compressing sterile non-micronized testosterone, anastrozole and stearic acid in a ratio of 80:4:2 into a 5 mm diameter cylindrical mold. The pellets are transferred to a sterile 5 ml amber vial covered with sterile cotton wool and sealed with a rubber stopper and a crimped aluminum tear seal. The vial is sterilized by gamma radiation at 25 KGy. USP grade non-micronized testosterone and anastrozole are obtained from Azelis (Brookvale, NSW). NF quality stearic acid is obtained from Medica (Mascot, NSW). The steps for producing these pellets are summarized as follows:

[0315] 1. Testosterone (non-micronized), anastrozole, and stearic acid were titrated by mechanical mixing for 2 hours.

[0316] 2. Mix thoroughly and titrate.

[0317] 3. The pellets were compressed by a press, and the hardness of the pellets produced was 8Kg / N.

[0318] 4. Store the pellets in amber glass 5 ml vials with sterile cotton wool pads.

[0319] 5. Gamma sterilize the stored pellets at 25KyG.

[0320] 6. Label the pellets correctly.

[0321] 7. Store in a cool, dry place (room temperature; 20-25°C, 66-78°F).

[0322] method

[0323] Premenopausal women with high mammographic breast density (MBD) were selected for the study. After successful screening, participants came to Wellend Clinic South Australia for dosing. Participants will be dosed during the luteal phase of their menstrual cycle to avoid the high changes in the hormonal environment during the follicular phase. After implant insertion, PK blood sampling will be performed for three months. Briefly, sampling will be repeated on the day of dosing, daily for the first week, weekly for the next four weeks, and every second week for the last eight weeks, for a total sampling period of three months. The trial ends after the assessment is completed on the last day of assessment, which is Day 85.

[0324] To determine whether the selected pharmacokinetic (PK) sampling protocol provided an adequate description of the T+Ai serum / plasma concentration-time profile, samples were analyzed for testosterone, dihydrotestosterone, and anastrozole after blood samples were collected from the first two participants on Day 29. The sampling protocol was modified by adding a blood sample (6 hours post-dose) and the timing of existing time points was not adjusted. The sampling duration was not extended.

[0325] In summary, blood samples were collected pre-dose (within one hour of dosing) and at 1, 2, 4, 6, 8, and 12 hours after dosing on Day 1. Participants returned at 9:00 AM on Days 2, 3, 4, 5, 8, 15, 22, 29, 43, 57, 71, and 85 for blood sampling.

[0326] A total of 11 subjects were dosed and included in the safety analysis set. Eleven subjects were included in the PK analysis set. Females were aged 35 to 55 years with a body mass index (BMI) range of 20-30 kg / m 2 The demographic data of the participants are shown in Table 1 .

[0327] Table 1: Summary of participant demographics (safety analysis set)

[0328] Demographic parameters (units) Number of participants average value Standard Deviation Median Minimum Maximum Gender (Female) 11 Age (years) 11 41.5 3.7 40 37 49 Weight(kg) 11 67.87 10.27 64.9 50 83.4 Height(m) 11 1.673 0.066 1.67 1.54 1.79 <![CDATA[BMI(kg / m 2 )]]> 11 24.14 2.26 23.4 20.8 27.4 Race (white) 10 Race (Caucasian / Asian) 1 Race (non-Hispanic or Latino) 11

[0329] PK results

[0330] The PK parameters of plasma anastrozole and serum testosterone are summarized in Table 2 and shown in Figures 1 to 6 .

[0331] Table 2: Summary of PK parameters of plasma anastrozole and serum testosterone

[0332]

[0333] Regarding pharmacokinetics, the median T of plasma anastrozole max The mean C max =5.30ng / mL, and the average terminal half-life was 14 days. After 12 weeks, the concentrations of most participants were below the limit of quantification (0.1ng / mL). The 14-day half-life observed from T+Ai is significantly longer than the 2-day half-life specified for oral anastrozole, confirming that the T+Ai product acts as a sustained-release formulation. Since the goal of this therapy is to reduce the aromatization of T at the insertion site (i.e., subcutaneous fat with average aromatase levels, and in tissues with high aromatase levels), the minimum effective dose is given compared to the treatment of breast cancer (where systemic aromatase inhibition is sought to maximize anti-tumor effects). The IC50 of anastrozole in tissues overexpressing aromatase (such as the MCF7 breast cancer cell line and the JEG3 human choriocarcinoma cell line) are 3.62 and 5.66nM, respectively. Therefore, C max =5.3 ng / mL (18.07 nM) will cover the initial higher C of T max =4.1 ng / mL and anastrozole is at a continuously lower level as T levels decrease. Thus, the use of 4 mg anastrozole in a pellet achieves the concentration required for aromatase IC50 sufficient to inhibit aromatase in target tissues. In contrast, a dose of 2 mg is insufficient.

[0334] For serum testosterone, the median T max The mean C max =4.1 ng / mL, compared with a mean of 0.2 ng / mL at baseline. A second lower peak occurred at weeks 2-3. Participants continued to have an increase in testosterone at week 12 compared with baseline, with a mean concentration of 0.8 ng / mL. Serum dihydrotestosterone was not measurable in most samples; in participants with a measurable maximum, T max This example shows that the pellets after subcutaneous insertion show patterns.

[0335] Example 2: Combination therapy of testosterone and anastrozole for reducing MBD in women

[0336] This example provides an analysis of the use of testosterone and anastrozole combination therapy to reduce mammographic breast density (MBD) in women. The study was conducted at the Wellend Clinic (Burnside War Memorial Hospital, Adelaide, South Australia). The primary indications for therapeutic intervention were one or more of the following: perimenopausal hormone dysfunction, high MBD, which is considered a factor in reducing breast cancer (BC) risk.

[0337] All 652 patients were female, with a mean age of 52 years (range, 23 to 79 years) at first T+Ai implantation.

[0338] MBD / BC risk reduction was the primary indication for treatment in 89 patients (14%), adjunctive with hormonal dysfunction in 334 patients (51%), and both indications in 177 patients (27%). No primary indication was provided for 52 patients.

[0339] A history of BC was noted in 90 patients (14% of patients had a non-missing response to BC history). The BC type was noted in 20 patients - 14 had in situ type BC and 6 had invasive type BC.

[0340] Concomitant estrogen-based medication use was noted in 222 patients (34%), the majority of which were topical forms (n=192).

[0341] At the July 2017 data cutoff, 365 patients (56%) were still continuing to receive T+Ai therapy. One patient was noted to have no disposition information, while the remaining 286 patients had discontinued T+Ai therapy. The most common reason for discontinuing T+Ai therapy was loss to follow-up (n=123), followed by the subject's decision to discontinue therapy (n=46), cost of therapy (n=30), the subject felt that T+Ai therapy was ineffective (n=28), and the primary care physician recommended that the patient's T+Ai therapy was completed (n=25).

[0342] Measurement of efficacy

[0343] Summary statistics for continuous variable measures, including %VBD and AVBD measures of MBD, were obtained from the mammograms. Because the mammograms were not scheduled for specific time points, an access window system was applied to the data in order to provide specific data values ​​for specific time points. The windows selected were as follows:

[0344] Six months (180 days + / - 60 days after first T+Ai implant)

[0345] One year (365 days + / - 180 days after first T+AI implant)

[0346] Two years (730 days + / - 180 days after first T+AI implant)

[0347] Three years (1095 days + / - 180 days after first T+AI implant)

[0348] Four years (1460 days + / - 180 days after first T+AI implant)

[0349] Relationship between drug dose and %VBD response

[0350] To examine the effect of the T+Ai intervention on MBD (measured by %VBD), a subset of patients who had mammograms before and after treatment initiation were considered. Change from baseline in MBD was used as the outcome measure in mixed model analyses in PRO CMIXED. This procedure allowed individual patients to contribute more than one mammogram after starting T+Ai treatment (repeated measures analysis) and examined the possibility of different covariance patterns between the data.

[0351] From the dataset provided for the analysis, the following independent (explanatory) variables were used to examine their effects on changes in MBD:

[0352] i. Days since first implantation

[0353] ii. Baseline %VBD MBD measurement (value closest to but not later than first T+Ai implant)

[0354] iii. Cumulative testosterone dose (mg) throughout the study (stratified as <500 mg, 500 mg to <700 mg, and 700 mg+).

[0355] iv. Cumulative anastrozole dose (mg) throughout the study (as a continuous covariate)

[0356] v. Age at first implantation (years)

[0357] vi. Machine type (GE or Hologic machine)

[0358] vii. Radiation Dose from Mammogram

[0359] viii. Compression pressure of mammogram

[0360] ix. History of breast cancer (yes or no)

[0361] x. Use of concomitant estrogen medication (yes or no)

[0362] xi. Interaction term between days since first implant and cumulative dose of testosterone.

[0363] The last interaction term listed above allows for different slope fits to be made for each testosterone dose stratum to see if there are potential differences between the strata.

[0364] The following syntax is used in SAS to fit the model:

[0365] proc mixed data = mmg3;

[0366] class TESTO_GPSUBJIDC MMG_MACH_N BCA_HIST_N E2_USED_N;

[0367] model CHANGE_MMG_RESULT=MMG_DAY BASE_MMG_RESULT TESTO_GP MMG_MACH_NBCA_HIST_N E2_USED_N SUM_ANAST MMG_RAD_DOSE_N MMG_COMP_PRES_N

[0368] MMG_DAY*TESTO_GP / ddfm=kr cl s;

[0369] random INT MMG_DAY / subject=SUBJIDC type=un;

[0370] lsmeans TESTO_GP / at MMG_DAY=365cl;

[0371] lsmeans TESTO_GP / at MMG_DAY=730cl;

[0372] lsmeans TESTO_GP / at MMG_DAY=1095cl;

[0373] lsmestimate TESTO_GP'Across all dose levels at 1yr'1 1 1 / divisor=3atMMG_DAY=365cl;

[0374] lsmestimate TESTO_GP'Across all dose levels at 2yr'1 1 1 / divisor=3atMMG_DAY=730cl;

[0375] lsmestimate TESTO_GP'Across all dose levels at 3yr'1 1 1 / divisor=3atMMG_DAY=1095cl;

[0376] format TESTO_GP testo.BCA_HIST_N E2_USED_N yesno.MMG_MACH_Nmachine;

[0377] run.

[0378] Each term in the model statement represents an item in the previous bulleted list. The random statement is used to account for repeated measurements within each patient over time. An unstructured covariance structure is used because It provided a slightly better fit when compared with the composite symmetry (CS) and autoregression (AR1) options in the LSMEANS and LSMESTIMATE statements. The LSMEANS and LSMESTIMATE statements were used to estimate the change from baseline in MBD by testosterone dose stratum at 1, 2, and 3 years. Table 3 shows the parameter estimates obtained from the model.

[0379] Note the statistically significant findings about the number of days since the first T+Ai implant, and the interaction between the number of days since the first T+Ai implant and the cumulative testosterone layer. Specifically, it was noted that patients with cumulative testosterone doses of more than 500 mg had a greater decrease in MBD over time compared to patients with <500 mg. The cumulative anastrozole dose also approached the significance level of 0.05 (p=0.06), and estimates also indicate that the higher the cumulative anastrozole dose, the more MBD is reduced. Baseline %VBD is also statistically significant, with the higher the baseline score, the greater the observed change (%VBD reduction).

[0380] Table 3: SAS PROC MIXED Model Estimates of Change from Baseline in MBD

[0381]

[0382] At 1 year, 2 years and 3 years, the %VBD of MBD was estimated by cumulative testosterone layer and the least squares mean of the change from baseline was measured. Table 4 below shows the results of the model. The highlighted row shows the estimated change from baseline, which is statistically significantly different from zero (p < 0.05). The least squares mean analysis of %VBD change from baseline was estimated at 1 year, 2 years and 3 years.

[0383] Table 4: Least Squares Estimation of Change from Baseline in MBD by Cumulative Testosterone Dose and Time Since First Implant

[0384] Cumulative testosterone dose group Time since first implantation Least Squares Mean Estimation Lower 95% CI Upper 95% CI p-value <500mg 1 year -1.6085 -2.8628 -0.3542 0.0123 <500mg 2 years -1.213 -2.6838 0.2579 0.1053 <500mg 3 years -0.8174 -2.8829 1.248 0.4349 500 to < 700 mg 1 year -1.1249 -2.4165 0.1668 0.0872 500 to < 700 mg 2 years -1.6916 -3.0057 -0.3775 0.0121 500 to < 700 mg 3 years -2.2584 -4.2275 -0.2893 0.0251 700+mg 1 year -1.8688 -3.6155 -0.1222 0.0362 700+mg 2 years -2.3358 -3.8817 -0.7899 0.0034 700+mg 3 years -2.8028 -4.6566 -0.949 0.0035

[0385] Efficacy of T+Ai in reducing MBD as measured by AVBD

[0386] In addition to the analysis performed using the %VBD MBD measurement, a supplemental analysis was performed using the same model discussed above in this example, with the change from baseline in absolute volumetric breast density (AVBD) as the dependent variable. The only other change to the list of dependent variables used was the replacement of baseline %VBD with baseline AVBD. The least squares mean estimates from the AVBD model are shown in Table 5 below. Again, statistically significant values ​​are shown in the highlighted rows. The only significant values ​​were those in the 700 mg + cumulative testosterone group, two and three years after the first T + Ai implant. The two-year and three-year AVBD change from baseline estimates were -22 cm 3 and -60cm 3 .

[0387] Table 5: Least Squares Estimation of Change from Baseline in AVBD by Cumulative Testosterone Dose and Time Since First Implant

[0388] Cumulative testosterone dose group Time since first implantation Least Squares Mean Estimation Lower 95% CI Upper 95% CI p-value <500mg 1 year -12.6001 -25.7233 0.523 0.0597 <500mg 2 years -11.2073 -28.3175 5.9029 0.1975 <500mg 3 years -9.8144 -35.6603 16.0314 0.4537 500 to < 700 mg 1 year 0.2747 -12.2756 12.8251 0.9652 500 to < 700 mg 2 years -4.1993 -19.8209 11.4223 0.595 500 to < 700 mg 3 years -8.6734 -34.0633 16.7166 0.4985 700+mg 1 year -7.7803 -26.2516 10.691 0.4052 700+mg 2 years -21.9964 -39.4828 -4.5101 0.0142 700+mg 3 years -36.2126 -59.7196 -12.7056 0.0029

[0389] Efficacy Conclusion

[0390] Using mammographic information obtained from 142 patients who had mammograms performed both before and after the start of T+Ai therapy, there were statistically significant changes from baseline in MBD measures for both %VBD and AVBD after T+Ai therapy intervention.

[0391] When MBD was measured by %VBD, the estimated change was -1.87 at 1 year after therapy initiation for patients with a cumulative testosterone dose of 700 mg+, -2.34 at 2 years after therapy initiation, and -2.82 at 3 years after therapy initiation. For patients with a cumulative testosterone dose of 500-<700 mg, the estimated change was 1.12 (p=0.09) at 1 year after therapy initiation, -1.69 at 2 years after therapy initiation, and -2.26 at 3 years after therapy initiation.

[0392] When MBD was measured by AVBD, significant values ​​were found in the 700 mg + cumulative testosterone group at 2 and 3 years after the first T + Ai implant. The estimated change from baseline AVBD at 2 and 3 years was -22 cm, respectively. 3 and -60cm 3 .

[0393] Most women in Example 2 received anastrozole at a dose of 2 mg per implant. This dose was chosen in an attempt to keep the dose as low as possible to avoid side effects. Although the efficacy of this dosing was effective, Example 2 demonstrated that cumulative anastrozole dosing was a variable of interest when assessing changes in MBD (p=0.06). Larger cumulative anastrozole dose values ​​were associated with greater changes from baseline (decreases from baseline) in MBD. Surprisingly and unexpectedly, the PK and PD of anastrozole shown in Example 1 demonstrated that larger doses of anastrozole (4 mg per implant) can be used that achieve efficacy and also avoid or minimize side effects associated with the aromatase inhibitors disclosed herein.

[0394] Example 3: Pharmacokinetic profile of anastrozole using a modeling approach

[0395] This example provides an analysis of the pharmacokinetic profile of anastrozole using a modeling approach to evaluate the optimal input function describing the absorption of the experiment described in Example 1. Table 6 lists the sampling schedule for anastrozole and testosterone. At least 8 mL of blood samples were collected by venipuncture at each time point specified in Table 6 for serum testosterone / DHT and plasma anastrozole. Fig. 7A -B illustrates the absorption rate of anastrozole. The data in these figures are expressed as mean + / - standard deviation. Fig. 7A The Y-axis is displayed on a logarithmic scale. Figure 7B The Y-axis is shown on a linear scale. This figure was generated using the pellet absorption data summarized in Table 2 of Example 1. The percent change from baseline in mean volume was calculated and plotted against time in months. Fig. 8A -B plots the plasma anastrozole concentrations after implantation. Fig. 8A In the data on the X-axis, the data are plotted as time (hours) after transplantation. Figure 8B In , the X-axis is on a logarithmic scale. Fig. 8A In -B, open circles equal the observed data, black line equals the median of the PK curve, and dark grey dashed line equals the lower limit of quantitation (0.1 ng / mL). Fig. 9 The observed frequencies after implantation are shown. Approximately 15% of the data were below the lower limit of quantitation (BLQ). BLQ concentrations were observed before dosing or 1000 hours after implantation.

[0396] Table 6

[0397]

[0398] In this example, a population modeling approach was used to characterize the PK of anastrozole. One- and two-compartment models were fitted to the data. Several absorption models were tested, such as first and zero order, mixed order, and transfer compartment. Model selection was performed based on statistical criteria (objective function) and visual inspection of diagnostic plots. Fig.10 A flow chart of the testing model is shown. Fig.11 The final structural model used in this example is shown. This example shows that the best model to fit the data is a two-compartment disposition model with absorption described by dual inputs. Inter-subject variability (BSV) is included in apparent clearance (CL / F), central compartment apparent distribution volume (Vc / F), and first-order absorption rate constant (KA). A proportional error model is used to describe the residual unexplained variability (RUV).

[0399] Table 7 below provides parameter estimates for anastrozole. Parameters were estimated with good precision (RSE < 30%) except for the inter-subject variability terms on Vc / F and apparent inter-compartmental clearance (Q / F), which, however, were considered acceptable given the available data. The model estimate of CL / F was 1.77 L / h, comparable to the NCA estimate of 1.88 L / h demonstrated in Table 2 of Example 1.

[0400] Table 7

[0401]

[0402]

[0403] Fig. 12A -B shows the group and individual predictions. Fig. 12A Group predictions are shown. Fig. 12B Individual forecasts are shown. The light grey line represents the identity line, and the dark grey line shows the data trend (Loess smoothing). Since the BLQ data is deleted, the forecast Fig. 12B The trend line will have tails. Overall, the model predicts the data well with minimal bias. The population predicted concentrations are the performance of a typical patient. The individual predicted concentrations are the posterior Bayes / individualized predictions. Fig.13 (ID:1-11) shows individual plots of observed and predicted anastrozole plasma concentrations. Data are plotted against nominal time. Black dashed lines equal population model predictions, gray solid lines equal individual model predictions, and circles equal individual observations. The final model was used to simulate 1000 replicates of the data set to validate the model simulation properties. 95% CIs around the median and 10 th and 90 th Prediction interval. Then the median of the observed data, 10 th and 90 th Percentiles are superimposed on simulated data to validate the model.

[0404] Fig.14Visual Predictive Check (VPC) of Anastrozole concentration after implantation is shown. The VPC shows that the observed data is well captured by the simulated data from the model. The X-axis and Y-axis are on a logarithmic scale. Black dashed line = observed 10 th and 90 th Percentiles, solid black line = observed median, grey shaded area = 80% prediction interval, light grey dashed line = lower limit of quantitation.

[0405] Fig.15A -B shows a one-compartment versus a two-compartment model. Fig.15A Indicates a primary single-room layout. Fig. 15B represents a two-compartment arrangement with two inputs. The distribution of the conditional weighted residual (CWRES) of the one-compartment model over time ( Fig.15A ) shows that there is misspecification in the elimination phase. In contrast, the two-compartment model ( Fig. 15B ) shows that the distribution of CWRES over time is uniform.

[0406] Fig.16 Predictions for patient 10 from several different input models are shown, including: single level input (model 9), simultaneous first and zero level input (model 10), single zero level input (model 11), and dual first level input (model 151). It is evident from the figure that models other than dual first level input fail to correctly capture the early stages of absorption.

[0407] Summarize

[0408] This analysis showed that anastrozole PK followed a two-compartment layout profile. The dual input described the absorption profile of anastrozole well. Other input models failed to capture the absorption phase correctly.

[0409] Example 4: Breast tissue elasticity

[0410] This example looks at the potential effects on breast tissue elasticity as assessed by shear wave ultrasound in subjects participating in the trial described in Example 1. Breast elasticity was measured by shear wave ultrasound measurements on days 1, 29, 57, and 85. Four measurements of glandular tissue elasticity and adipose tissue elasticity were taken for each breast. The calculated average of the eight measurements for each tissue type was the value used to summarize and analyze breast elasticity. A summary of the breast elasticity at each time point and the change from baseline is provided in Table 8. Table 8 shows that breast tissue elasticity gradually decreased over time, with glandular tissue decreasing 28% from baseline and adipose tissue decreasing 32% from baseline.

[0411] Table 8

[0412]

[0413] Example 6: Expression of CD36

[0414] The breast tissue of women with autoimmune inflammatory mastitis (AIM) has high aromatase and estradiol, so the expression of Tregs is severely suppressed. Inducing CD36 in the breast tissue of women with AIM may lead to tissue-specific immunosuppression rather than systemic immunosuppression. This example shows that CD36 can be expressed in normal breast tissue by treating with a combination of androgen agents and aromatase inhibitors as described herein.

[0415] Normal breast tissue was removed from 3 perimenopausal women obtained during surgery. Tissue samples were transported to the laboratory on ice, and the maximum time between excision and tissue processing was one hour. 5ml 200mM glutamine (SAFCbiosciences, Kansas, USA), 5ml 100x antibiotic / antimycotic (Sigma, St Louis, MO, USA), 10 μg / ml insulin (Sigma, St Louis, MO, USA) and 10 μg / ml hydrocortisone (Sigma, St Louis, MO, USA) culture medium (phenol red-free RPMI; SAFC biosciences, Kansas, USA) were washed in breast tissue samples to remove excess blood.

[0416] A representative block of each tissue sample was immediately fixed in 4% formalin in phosphate buffered saline (PBS) at 4°C overnight, then dehydrated using an automated tissue processor (Sakura Tissue-Tek VIP, USA) and embedded in solid paraffin. Hematoxylin and eosin (H&E) stained sections were used to evaluate histopathology. The remaining fresh tissue was cut into small pieces (~3x3x1 mm) and placed in triplicate on 1 cm 3 Gel sponges (Spongostan; Johnson & Johnson, Skipton, UK) were pre-soaked and then half-immersed in treatment medium containing 10% dextran-coated charcoal treated fetal calf serum (DCC-FCS) (SAFC biosciences, Kansas, USA) in a 24-well tissue culture plate (BD Biosciences, NJ, USA). Tissue samples were then cultured in a vehicle containing 0.1% ethanol, 5 nM testosterone and 25 ng / ml anastrozole for 24 hours.

[0417] Western blot whole cell lysate of control adipose tissue was prepared by sonication at 48°C in lysis buffer (1% Triton X-100, 50mM KCl, 25mM HEPES, pH 7.8, 10mg / ml leupeptin, 20mg / ml aprotinin, 125mM dithiothreitol and 1mM phenylmethylsulfonyl fluoride) and analyzed on the same western blot. 50mg total protein sample was mixed with 50ml sodium dodecyl sulfate (SDS)-mercaptoethanol sample buffer and boiled for 10 minutes, then protein was separated on 7.5% SDS gel and transferred to polyvinylidene difluoride membrane. The membrane was then blocked with 5% skim milk in phosphate buffered saline (PBS) containing 0.5% Tween-20 for 1 hour at room temperature and immunoblotted with an antibody against human CD36 diluted in PBS and a horseradish peroxidase-conjugated secondary antibody (Jackson Immunoresearch) diluted in PBS, followed by detection with a chemiluminescent reagent (Amersham Bioscience, Buckinghamshire, England). Band density was measured by densitometry using Image Master VDS and Image Quant Analysis Software (Amersham Pharmacia Biotech, Hong Kong, China). Relative protein levels of CD36 and β-actin in the original total protein lysate from the breast product were obtained. CD36 protein expression was normalized to β-actin expression. Antibodies containing synthetic peptides derived from the human CD36 sequence were produced in the immunized host, purified by peptide affinity chromatography and confirmed using a control peptide. Fig.17 Results of western blot analysis of CD36 protein in 3 explant samples at baseline and after 24 hours of culture are shown. Cell lysates were immunoblotted with an antibody against CD36. The experiment was performed twice with similar results. Data are expressed relative to actin. A 2-tailed Student's t-test showed a significance of p=0.00757. Results are listed in Table 9 below.

[0418] Table 9: Western blot band density measurements

[0419] Number of patients Before treatment After treatment 1 0.3235 0.9845 2 0.2135 1.0156 3 0.1478 0.4875 average value 0.2283 0.8292 SC 0.0888 0.29634

[0420] Increasing CD36 increases the conversion of fibroblasts to adipocytes (fat). Adipocytes (fat) are semi-fluid and therefore have greater elasticity. This example shows that CD36 in normal breast tissue increases significantly after combined treatment with an androgen testosterone and an aromatase inhibitor anastrozole. Increased CD36 expression is associated with reduced breast firmness. As described herein, certain embodiments of the present disclosure relate to combined therapy using an androgen and an aromatase inhibitor to increase CD36 and reduce breast firmness.

[0421] The above demonstrates that in normal breast tissue, the combination of testosterone and an aromatase inhibitor is very effective in inducing CD36, as demonstrated by the Western blot below, which shows that CD36 is largely induced by treatment of normal breast tissue harvested during surgery and grown in explants.

[0422] Example 7: Idiopathic Inflammatory Macromastia

[0423] As discussed below, four patients with idiopathic inflammatory mastitis associated with mild to moderate macromastia were treated according to the present disclosure.

[0424] Patient 1 :42.03416971

[0425] Medical consultation

[0426] A 38-year-old chiropractor presented with rapid breast enlargement—"doubling in size," periareolar inflammation, and persistent severe pain over a 2-month period that was unresponsive to over-the-counter analgesics and NSAIDs and affected the patient's ability to work.

[0427] Bra cup size before onset

[0428] The patient's bra cup size is 32A and she has been unable to wear a bra since the onset of the disease.

[0429] Past medical history

[0430] The patient had severe premenstrual dysphoric disorder partially controlled by oral contraceptives, which she had stopped taking 2 years before presentation, with regular 28-day cycles, 2 pregnancies, and 2 live births. Severe postpartum arthritis—undiagnosed—resolved 1 year later, and no abnormal blood parameters were noted.

[0431] VAS pain scale before onset: 95mm

[0432] treat

[0433] Subcutaneous implantation of pellets of 80 mg testosterone and 4 mg anastrozole as described in Example 1 for 3 months, repeated for two additional 3 month periods

[0434] result

[0435] i. VAS pain scale 10mm at 4 weeks

[0436] ii. No inflammation detected at 4 weeks

[0437] iii. Fibroglandular tissue decreased by 71%

[0438] iv. Breast volume decreased by 58% and returned to the size before the onset of the disease

[0439] v. Complete reversal of extreme background parenchymal enhancement on MRI, sustained for 3 years after cessation of therapy

[0440] vi. No SAE

[0441] vii. No change in BMI

[0442] Mammograms of the patient's breasts before and after treatment Fig.19 A, and MRI images before and 3 years after treatment are shown in Fig.19 C. Breast volume and density measurements are shown in Fig.19 As shown in B.

[0443] Patient 2: 42.09761311

[0444] Medical consultation

[0445] A 42-year-old nurse with rapidly enlarging breasts—“very hard,” diffuse inflammation, persistent severe pain over 4 months, unresponsive to over-the-counter analgesics and NSAIDs, affecting work and family life—could not ride her horse. The left breast was much larger than the right.

[0446] Bra cup size: before onset

[0447] The patient's bra cup size was 36C and she was unable to wear a bra after the onset of the disease. She wore a sports bra all day and wore it to bed.

[0448] Past medical history

[0449] Regular 28-day cycles, 2 pregnancies and 2 live births with severe preeclampsia, postpartum depression including hospitalization, no abnormal blood parameters noted.

[0450] VAS pain scale before onset: 100mm

[0451] treat

[0452] Subcutaneous pellets of 80 mg testosterone and 4 mg anastrozole were implanted for 3 months and repeated for two additional 3-month periods.

[0453] result

[0454] i. VAS pain scale 15 mm at 4 weeks

[0455] ii. No inflammation detected at 4 weeks

[0456] iii. Fibroglandular tissue decreased by 41%

[0457] iv. Breast volume decreases by 10% and returns to pre-onset size

[0458] v.No SAE

[0459] vi. No change in BMI

[0460] vii. No mammoplasty

[0461] Mammograms of the patient's breasts before and after treatment Fig. 20 A. Breast volume and density measurements are as follows Fig. 20 As shown in B.

[0462] Patient 3: 42.09761311

[0463] Medical consultation

[0464] A 37-year-old attorney with rapidly enlarging breasts—inflammation, especially around the nipple, and persistent, severe pain over a 2-month period that was unresponsive to over-the-counter analgesics and NSAIDs—interfered with work and family life.

[0465] Bra cup size

[0466] 34C before onset, unable to wear a bra after onset, wear sports bra all day, and wear sports bra to sleep

[0467] Past medical history

[0468] Regular 28-day cycles, no pregnancies, moderate endometriosis, no abnormal blood parameters noted.

[0469] VAS pain scale before onset: 100mm

[0470] treat

[0471] Subcutaneous implants of 80 mg testosterone and 4 mg anastrozole as described in Example 1 were continued for 3 months and repeated for two additional 3 month periods.

[0472] result

[0473] i. VAS pain scale 0 mm at 4 weeks

[0474] ii. No inflammation detected at 4 weeks

[0475] iii. Fibroglandular tissue decreased by 52%

[0476] iv. Breast volume decreased by 32% and returned to pre-onset size

[0477] v.No SAE

[0478] vi. No change in BMI

[0479] vii. No mammoplasty

[0480] Mammograms of the patient's breasts before and after treatment Fig.21 A. Breast volume and density measurements are as follows Fig.21 As shown in B.

[0481] Patient 4: 42.83593371

[0482] Medical consultation

[0483] A 41-year-old police officer with rapidly enlarging breasts—“very hard,” diffuse inflammation, and persistent severe pain for 1 month that was unresponsive to over-the-counter analgesics and NSAIDs, affecting the patient’s work and family life—was unable to wear the Kevlar protective kit.

[0484] Bra cup size

[0485] 36B before the onset of the disease, unable to wear a bra after the onset of the disease, wear a sports bra all day, and wear a sports bra to sleep

[0486] Past medical history

[0487] Regular 28 day cycles, no pregnancy, severe endometriosis, multiple surgeries, no abnormal blood parameters noted.

[0488] VAS pain scale before onset: 100mm

[0489] treat

[0490] Subcutaneous implants of 80 mg testosterone and 4 mg anastrozole as described in Example 1 were continued for 3 months, repeated for two additional 3 month periods, mammoplasty, and then two additional 3 month periods of the above post-operative treatment.

[0491] result

[0492] i. VAS pain scale 0 mm at 4 weeks

[0493] ii. No inflammation detected at 4 weeks

[0494] iii. Fibroglandular tissue decreased by 36%

[0495] iv. Breast volume decreased by 23% and returned to pre-onset size

[0496] v. Complete reversal of extreme background parenchymal enhancement on MRI

[0497] vi. No SAE

[0498] vii. No change in BMI

[0499] MRI images of the patient's breast before and after treatment Fig. 22 A, and mammographic images of the patient's breast before and after treatment are shown in Fig. 22 B. Breast volume and density measurements are as follows Fig. 22 As shown in C.

[0500] Example 8: Treatment of autoimmune inflammatory mastitis

[0501] patient: 42.04033771

[0502] Medical consultation

[0503] A 24-year-old physiotherapist with rapidly enlarging breasts - "extremely painful", diffuse inflammation and constant severe pain over a 4-month period - unable to work and unresponsive to:

[0504] i. Over-the-counter analgesics and NSAIDs

[0505] ii.OCP

[0506] iii. Oral progesterone

[0507] iv. Danazol

[0508] v. LhRh agonists

[0509] Bra cup size

[0510] 36B before onset, increased to 36EE

[0511] Past medical history

[0512] Obstetric / gynecological history included irregular 28-day cycles-amenorrhea due to Zoladex, no pregnancies. No abnormal blood parameters were noted.

[0513] VAS pain scale before onset: 100mm

[0514] treat

[0515] Subcutaneous implants of 100 mg testosterone and 3 mg anastrozole pellets were continued for 3 months, followed by reduction mammoplasty, and then another two of the above postoperative treatments for 3 months.

[0516] result

[0517] i. VAS pain scale 50 mm at 4 weeks

[0518] ii. Significant reduction in inflammation at 4 weeks

[0519] iii. Reversal of mammographic hyperdensity and MRI extreme background parenchymal enhancement

[0520] iv. No SAE

[0521] v. BMI did not change

[0522] vi. Later diagnosed with myasthenia gravis

[0523] Mammographic images of the patient's breast before treatment Fig.23 A. The MRI image after treatment is shown in Fig.23 As shown in B.

[0524] Example 9: Treatment of plasma cell mastitis

[0525] Plasma cell mastitis is an autoimmune inflammatory destructive process of the milk ducts behind the areola, resulting in multiple fistulas, and inevitably leading to disfiguring surgery and a high risk of recurrence. Except for surgery with significant limitations, there is no known treatment for this condition. An example of plasma cell mastitis is that the milk duct system is invaded by inflammatory cells that secrete proinflammatory cytokines. It has recently been shown that the IL-6 inflammatory pathway is crucial in this inflammatory process and has been targeted as a potential mechanism for the treatment of plasma cell mastitis (Liu, 2020). It has been shown that female breasts have cells that respond violently (reduced by 53%) to testosterone treatment in reducing IL-6 levels (Guhl, 2012).

[0526] A 43-year-old otherwise healthy woman developed multiple fistulas in the right nipple region of the complex that initially did not respond to antibiotic therapy and then high-dose corticosteroids and total ductectomy as a surgical procedure with only short-term relief. There was significant inflammation around the right nipple-areola complex with four fistulas at the 4 o'clock position at the areola margin. Significant pain was measured at 8 on a 0 to 10 cm visual analog pain scale. She started T+AI therapy and received a subcutaneous implant of 80 mg testosterone plus 4 mg anastrozole. Within three weeks, her visual analog pain scale had decreased to 3 and there was also a decrease in the periareolar redness. Eleven months after insertion of three implants of the same concentration, the fistula formation had completely resolved with no associated pain. Follow-up 12 months later showed no evidence of recurrent disease.

[0527] Example 10: Granulomatous Mastitis

[0528] Granulomatous mastitis is also called granulomatous lobular mastitis because the inflammation occurs around the lobules of the breast. It is a manifestation of autoimmune inflammatory mastitis in the breast tissue, resulting in granuloma formation, inflammation, and fistula formation. There is no known cure for this condition, and women often undergo multiple surgeries and / or receive immunosuppressive therapy.

[0529] A 32-year-old woman presented for a second opinion 6 months after being treated for histologically confirmed granulomatous mastitis. She had been given antibiotics, corticosteroids, methotrexate, and had surgery to remove the fistula. All of these had failed, as evidenced by an MRI taken on July 23, 2017, which showed multiple areas of granuloma formation and a large reactive axillary lymph node.

[0530] Between the first MRI and the second MRI taken on May 23, 2018, she had three testosterone 80 mg anastrozole 4 mg implants inserted as described in Example 1. The inflammation decreased rapidly and the one remaining fistula healed over a three-month period. Tenderness and breast discomfort persisted for five months and then slowly subsided. A clinical examination on May 23, 2018 showed only a slight scar on the breast from the previous surgery, but no other abnormalities. When re-examined in February 2020, she remained free of disease.

[0531] Mammographic images of the patient's breast before treatment Fig.24 A. The MRI images obtained after treatment are shown in Fig.23 As shown in B.

[0532] Other exemplary non-limiting embodiments.

[0533] Further advantages of the claimed subject matter will become apparent from the following examples describing certain embodiments of the claimed subject matter.

[0534] A Example

[0535] 1A. A pharmaceutical preparation comprising:

[0536] an effective amount of an androgen, an effective amount of an aromatase inhibitor, and a binding agent;

[0537] The formulation, when administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by serum concentrations of the androgen and plasma concentrations of the aromatase inhibitor; and

[0538] The sustained-release multiphasic concentration pattern in the subject's serum or plasma includes:

[0539] a first time period in which the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and

[0540] A second time period wherein the androgen initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

[0541] 2A. A pharmaceutical preparation comprising:

[0542] an effective amount of an androgen, an effective amount of an aromatase inhibitor, and a binding agent;

[0543] The pharmaceutical preparation is compressed into pellets;

[0544] The pellet, when subcutaneously administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by serum concentration of the androgen and plasma concentration of the aromatase inhibitor; and

[0545] The sustained-release multiphasic concentration pattern in the subject's serum or plasma includes:

[0546] a first time period in which the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and

[0547] A second time period wherein the androgen initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

[0548] 3A. A pharmaceutical preparation comprising:

[0549] an effective amount of an androgen, an effective amount of an aromatase inhibitor, and a binding agent;

[0550] The formulation, when administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by serum concentrations of the androgen and plasma concentrations of the aromatase inhibitor; and

[0551] The sustained-release multiphasic concentration pattern in the subject's serum or plasma includes:

[0552] a first time period in which the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma;

[0553] a second time period wherein the androgen initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma;

[0554] a third time period, wherein the androgen has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the androgen during the third time period; and

[0555] The fourth time period, wherein the serum concentration level of the androgen gradually decreases, and the concentration of the aromatase inhibitor in the plasma gradually decreases, and the decrease levels of the two are approximately the same.

[0556] 4A. A pharmaceutical preparation comprising:

[0557] an effective amount of an androgen, an effective amount of an aromatase inhibitor, and a binding agent;

[0558] The pharmaceutical preparation is compressed into pellets;

[0559] The pellet, when subcutaneously administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by serum concentration of the androgen and plasma concentration of the aromatase inhibitor; and

[0560] The sustained-release multiphasic concentration pattern in the subject's serum or plasma includes:

[0561] a first time period in which the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma;

[0562] a second time period wherein the androgen initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma;

[0563] a third time period, wherein the androgen has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the androgen during the third time period; and

[0564] The fourth time period, wherein the serum concentration level of the androgen gradually decreases, and the concentration of the aromatase inhibitor in the plasma gradually decreases, and the decrease levels of the two are approximately the same.

[0565] 5A. A pharmaceutical preparation comprising:

[0566] 60mg to 120mg testosterone or its esters, 4mg to 6mg aromatase inhibitor and stearic acid;

[0567] The drug formulation is compressed into pellets having a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm;

[0568] The pellet, when subcutaneously administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time as measured by serum concentration of the testosterone or ester thereof and plasma concentration of the aromatase inhibitor; and

[0569] The sustained release multi-phase concentration mode includes:

[0570] a first time period wherein the testosterone or ester thereof has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and

[0571] A second time period wherein the testosterone or ester thereof initially has a decreased serum concentration level and then has an increased serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

[0572] 6A. A pharmaceutical preparation comprising:

[0573] 60mg to 120mg testosterone or its esters, 4mg to 6mg aromatase inhibitor and stearic acid;

[0574] The drug formulation is compressed into pellets having a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm;

[0575] The pellet, when subcutaneously administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time as measured by serum concentration of the testosterone or ester thereof and plasma concentration of the aromatase inhibitor; and

[0576] The sustained release multi-phase concentration mode includes:

[0577] a first time period wherein the testosterone or ester thereof has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma;

[0578] a second time period wherein the testosterone or ester thereof initially has a decreased serum concentration level and then has an increased serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma;

[0579] a third time period, wherein the testosterone or its ester has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the testosterone or its ester during the third time period; and

[0580] A fourth time period, wherein the serum concentration level of testosterone or its ester gradually decreases, and the concentration of the aromatase inhibitor in plasma gradually decreases, and the decrease levels of the two are approximately the same as each other.

[0581] 7A. The pharmaceutical formulation of example 1A, 2A or 5A, wherein the sustained release multiphasic concentration profile further comprises:

[0582] a third time period, wherein the testosterone or its ester has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the testosterone or its ester during the third time period; and

[0583] A fourth time period, wherein the serum concentration level of testosterone or its ester gradually decreases, and the concentration of the aromatase inhibitor in plasma gradually decreases, and the decrease levels of the two are approximately the same as each other.

[0584] 8A. The pharmaceutical formulation of one or more of Examples 1A to 7A, wherein during the first time period, the aromatase inhibitor exhibits a first order release.

[0585] 9A. The pharmaceutical formulation of one or more of Examples 1A to 8A,

[0586] Wherein during the first time period, the aromatase inhibitor does not exhibit zero-order release.

[0587] 10A. The pharmaceutical formulation of one or more of Examples 1A to 9A, wherein during the second time period, the aromatase inhibitor does not exhibit zero order release.

[0588] 11A. The pharmaceutical formulation of one or more of Examples 1A to 10A, wherein during the second time period, the testosterone or ester thereof does not exhibit zero order release.

[0589] 12A. The pharmaceutical formulation of one or more of Examples 1A-11A, wherein during the third time period, the aromatase inhibitor does not exhibit zero order release.

[0590] 13A. The pharmaceutical formulation of one or more of Examples 1A to 12A, wherein during the third time period, the testosterone or ester thereof does not exhibit zero order release.

[0591] 14A. The pharmaceutical formulation of one or more of Examples 1A to 13A, wherein during the third time period, the aromatase inhibitor exhibits a first order release.

[0592] 15A. The pharmaceutical formulation of one or more of Examples 1A to 14A, wherein during the third time period, the testosterone or ester thereof exhibits a first order release.

[0593] 16A. The pharmaceutical formulation of one or more of Examples 1A to 15A, wherein during the fourth time period, the aromatase inhibitor does not exhibit zero order release.

[0594] 17A. The pharmaceutical formulation of one or more of Examples 1A to 16A, wherein during the fourth time period, the testosterone or ester thereof does not exhibit zero order release.

[0595] 18A. The pharmaceutical formulation of one or more of Examples 1A to 17A, wherein during the fourth time period, the aromatase inhibitor exhibits a first order release.

[0596] 19A. The pharmaceutical formulation of one or more of Examples 1A to 18A, wherein during the fourth time period, the testosterone or ester thereof exhibits a first order release.

[0597] 20A. The pharmaceutical formulation of one or more of Examples 1A to 19A, wherein the first time period ends immediately after the androgen has a first peak concentration (Tmax) in serum.

[0598] 21A. The pharmaceutical formulation of one or more of Examples 1A to 20A, wherein the first time period ends at 5 hours to 14 hours.

[0599] 22A. The pharmaceutical formulation of one or more of Examples 1A to 21A, wherein the first time period ends at 5.5 hours to 13 hours.

[0600] 23A. The pharmaceutical formulation of one or more of Examples 1A to 22A, wherein the second time period ends immediately after the aromatase inhibitor has its Tmax.

[0601] 24A. The pharmaceutical formulation of one or more of Examples 1A to 23A, wherein the second time period ends at 23 hours to 80 hours.

[0602] 25A. The pharmaceutical formulation of one or more of Examples 1A to 24A, wherein the pharmaceutical formulation is an implant.

[0603] 26A. The pharmaceutical formulation of one or more of Examples 1A to 25A, wherein the implant is a compressed pellet.

[0604] 27A. The pharmaceutical formulation of one or more of Examples 1A to 24A, wherein the pharmaceutical formulation is a transdermal patch.

[0605] 28A. The pharmaceutical formulation of one or more of Examples 1A to 25A, wherein the implant is administered subcutaneously to a subject.

[0606] 28A. The pharmaceutical formulation of one or more of Examples 1A to 26A, wherein the compressed pellet is administered subcutaneously to a subject.

[0607] B Example

[0608] 1B. A pharmaceutical preparation, comprising:

[0609] about 80 mg testosterone or its ester, about 4 mg anastrozole, and about 2 mg stearic acid;

[0610] The drug formulation is compressed into pellets having a diameter of 4.4 mm to 4.6 mm and a length of 4 mm to 7 mm;

[0611] The pellet, when subcutaneously administered to a subject, provides a sustained-release multiphasic concentration pattern in the subject's blood over time, as measured by serum concentrations of the testosterone or ester thereof and plasma concentrations of the anastrozole; and

[0612] The sustained release multi-phase concentration mode includes:

[0613] a first time period, wherein the testosterone or ester thereof has a first peak concentration (Tmax) in serum and the concentration of anastrozole in plasma increases but is below its Tmax concentration in plasma;

[0614] a second time period wherein the testosterone or ester thereof initially has a decreased serum concentration level and then has an increased serum concentration level and the anastrozole has its Tmax concentration in plasma;

[0615] a third time period, wherein the testosterone or its ester has a second peak concentration in serum that is lower than Tmax, and the concentration of anastrozole in plasma gradually decreases and falls below the concentration of testosterone or its ester during the third time period; and

[0616] The fourth time period, wherein the serum concentration level of the testosterone or its ester gradually decreases, and the plasma concentration of the anastrozole gradually decreases, and the decrease levels of the two are approximately the same.

[0617] Although certain embodiments have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. The following claims are intended to define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

[0618] In the above description of certain embodiments, specific terms have been used for the sake of clarity. However, the present disclosure is not intended to be limited to the specific terms so selected, and it should be understood that each specific term includes other technical equivalents that operate in a similar manner to achieve similar technical purposes. The use of terms such as "left" and "right", "front" and "back", "upper" and "lower" is for convenience to provide a reference point and should not be interpreted as limiting terms.

[0619] In this specification, the word "comprising" is to be understood in its "open" sense, i.e. in the sense of "including", and is therefore not limited to its "closed" sense, i.e. in the sense of "consisting only of". A corresponding meaning is to be attributed to the corresponding words "comprise", "comprised" and "comprises" where they appear.

[0620] Additionally, the above describes only some embodiments of the present invention, and changes, modifications, additions and / or variations may be made thereto without departing from the scope and spirit of the disclosed embodiments, which are intended to be illustrative rather than restrictive.

[0621] It should be understood that the present invention is not limited to the disclosed embodiments, but on the contrary, is intended to cover various modifications and equivalent arrangements included in the spirit and scope of the present invention. In addition, the various embodiments described above can be implemented in combination with other embodiments, for example, aspects of one embodiment can be combined with aspects of another embodiment to implement other embodiments. In addition, each independent feature or component of any given component can constitute an additional embodiment.

Claims

1. A pharmaceutical preparation, comprising: an effective amount of an androgenic agent, an effective amount of an aromatase inhibitor, and a combining agent; The pharmaceutical preparation is compressed into pellets; The pellet, when subcutaneously administered to a patient, provides a sustained-release multiphasic concentration pattern in the patient's blood over time, as measured by serum concentrations of the androgenic agent and plasma concentrations of the aromatase inhibitor; and The sustained release multiphasic concentration pattern in the patient's serum or plasma includes: a first time period in which the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and A second time period wherein the androgen agent initially has a decreasing serum concentration level and then has an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

2. The pharmaceutical formulation according to claim 1, wherein the sustained release multiphasic concentration pattern comprises one or more of the following: During said first time period, said aromatase inhibitor exhibits a first order release, during said first time period, said aromatase inhibitor does not exhibit a zero order release, during the second time period, the aromatase inhibitor does not exhibit a zero order release, During the second time period, the androgenic agent does not exhibit zero order release.

3. The pharmaceutical formulation according to claim 1 or 2, wherein the sustained release multi-phase concentration pattern further comprises: a third time period, wherein the androgen agent has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the testosterone or its ester during the third time period; and The fourth time period, wherein the serum concentration level of the androgen agent gradually decreases, and the concentration of the aromatase inhibitor in plasma gradually decreases, and the decrease levels of the two are approximately the same as each other.

4. The pharmaceutical formulation according to claim 3, wherein the sustained release multiphasic concentration pattern comprises one or more of the following: During the third time period, the aromatase inhibitor does not exhibit zero order release, During the third time period, the androgenic agent does not exhibit a zero order release, During the third time period, the aromatase inhibitor exhibits a first order release, During the third time period, the androgenic agent exhibits a first order release, During the fourth time period, the aromatase inhibitor does not exhibit zero order release, During the fourth time period, the androgenic agent does not exhibit a zero order release, During the fourth time period, the aromatase inhibitor exhibits a first order release, During the fourth time period, the androgenic agent exhibits a first order release.

5. The pharmaceutical formulation according to any one of claims 1 to 4, wherein the first time period ends immediately after the androgen agent has a first peak concentration (Tmax) in serum.

6. The pharmaceutical formulation according to any one of claims 1 to 5, wherein the first period of time ends between 5 hours and 14 hours, or preferably, the first period of time ends between 5.5 hours and 13 hours.

7. The pharmaceutical formulation according to any one of claims 1 to 6, wherein the second time period ends immediately after the aromatase inhibitor has its Tmax.

8. The pharmaceutical formulation according to any one of claims 1 to 7, wherein the second period of time ends at 23 hours to 80 hours.

9. The pharmaceutical formulation according to any one of claims 1 to 8, wherein the androgenic agent is testosterone or a pharmaceutically acceptable salt or ester thereof.

10. The pharmaceutical preparation according to claim 9, wherein the androgen agent is selected from the group consisting of testosterone, methyltestosterone, dehydroepiandrosterone and testosterone undecanoate, preferably testosterone.

11. The pharmaceutical formulation according to any one of claims 1 to 10, for subcutaneous delivery of the androgen agent and the aromatase inhibitor.

12. A pharmaceutical preparation, comprising: 60 mg to 120 mg testosterone or an ester thereof, 2 mg to 6 mg aromatase inhibitor and more preferably 4 mg to 6 mg aromatase inhibitor, and stearic acid; The drug formulation is compressed into pellets having a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm; said pellet, when administered subcutaneously to a patient, provides a sustained-release multiphasic concentration pattern in the patient's blood over time as measured by serum concentration of said testosterone or ester thereof and plasma concentration of said aromatase inhibitor; and The sustained release multi-phase concentration mode includes: a first time period wherein the testosterone or ester thereof has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; and A second time period wherein the testosterone or ester thereof initially has a decreased serum concentration level and then has an increased serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma.

13. A pharmaceutical preparation, comprising: 60 mg to 120 mg testosterone or an ester thereof, 2 mg to 6 mg aromatase inhibitor and more preferably 4 mg to 6 mg aromatase inhibitor, and stearic acid; The drug formulation is compressed into pellets having a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm; said pellet, when administered subcutaneously to a patient, provides a sustained-release multiphasic concentration pattern in the patient's blood over time as measured by serum concentration of said testosterone or ester thereof and plasma concentration of said aromatase inhibitor; and The sustained release multi-phase concentration mode includes: a first time period wherein the testosterone or ester thereof has a first peak concentration (Tmax) in serum and the aromatase inhibitor increases in plasma concentration but is below its Tmax concentration in plasma; a second time period wherein the testosterone or ester thereof initially has a decreased serum concentration level and then has an increased serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma; a third time period, wherein the testosterone or its ester has a second peak concentration in serum that is lower than Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases and falls below the concentration of the testosterone or its ester during the third time period; and A fourth time period, wherein the serum concentration level of testosterone or its ester gradually decreases, and the concentration of the aromatase inhibitor in plasma gradually decreases, and the decrease levels of the two are approximately the same as each other.

14. The pharmaceutical preparation according to any one of claims 1 to 13, wherein the aromatase inhibitor blocks the conversion of androgens into estrogens under the action of aromatase.

15. The pharmaceutical formulation according to any one of claims 1 to 14, wherein the aromatase inhibitor is selected from the group consisting of anastrozole, exemestane and letrozole, preferably anastrozole.

16. The pharmaceutical formulation according to any one of claims 1 to 15, wherein the hardness of the pellets ranges from about 6 Kg / N to about 10 Kg / N, preferably about 8 Kg / N, so as to provide a multi-phase release pattern for the pellets.

17. Use of a pharmaceutical formulation according to any one of claims 1 to 16 for treating a condition in a patient selected from the group selected from: high mammographic breast density, breast pain, breast stiffness, macromastia, endometriosis, breast inflammation, gynecomastia, autoimmune inflammatory mastitis and breast cysts.

18. Use of an androgenic agent in combination with an effective amount of an aromatase inhibitor in the preparation of a medicament for preventing or treating autoimmune inflammatory mastitis in a patient in need thereof.

19. Use of an aromatase inhibitor in combination with an effective amount of an androgenic agent in the preparation of a medicament for preventing or treating autoimmune inflammatory mastitis in a patient in need thereof.

20. The use according to claim 18 or 19, wherein the androgenic agent and the aromatase inhibitor are used in the same medicament.

21. The use according to claim 20, wherein the drug is a sustained release drug formulation.

22. The use according to claim 21, wherein the medicament is in a solid dosage form for subcutaneous delivery to a patient.

23. The use according to claim 22, wherein the drug is in the form of pellets.

24. The use according to any one of claims 20 to 23, wherein the medicament provides a sustained-release multiphasic concentration pattern in the patient's blood over time as measured by serum concentration of the androgen and plasma concentration of the aromatase inhibitor.

25. The method according to claim 24, wherein the medicament is a pharmaceutical formulation as defined in any one of claims 1 to 16.

26. The use according to any one of claims 18 to 25, wherein the androgenic agent is testosterone or a pharmaceutically acceptable salt or ester thereof.

27. The use according to claim 26, wherein the androgenic agent is selected from the group consisting of testosterone, methyltestosterone, dehydroepiandrosterone and testosterone undecanoate.

28. The use according to claim 27, wherein the androgenic agent is testosterone.

29. The use according to any one of claims 18 to 28, wherein the aromatase inhibitor blocks the conversion of androgens into estrogens by the action of aromatase.

30. Use according to any one of claims 18 to 29, wherein the aromatase inhibitor is selected from the group consisting of anastrozole, exemestane and letrozole, preferably anastrozole.

31. The use according to any one of claims 18 to 30, wherein the autoimmune inflammatory mastitis is selected from the group consisting of idiopathic inflammatory macromastitis, plasma cell mastitis, granulomatous mastitis, and combinations of the foregoing.

32. The use according to claim 31, wherein the autoimmune inflammatory mastitis is granulomatous mastitis or plasma cell mastitis.

33. The use according to any one of claims 18 to 32, wherein the hardness of the pellets ranges from about 6 Kg / N to about 10 Kg / N, preferably about 8 Kg / N, so as to provide a multi-phase release pattern for the pellets.

34. The pharmaceutical formulation according to any one of claims 1 to 17 or the use according to any one of claims 18 to 33, wherein the patient is a female patient, preferably selected from the group consisting of perimenopausal women, menopausal women or postmenopausal women.

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