Pharmaceutical formulations and systems for delivering androgenic agents and aromatase inhibitors and methods of use

Abstract

The present disclosure relates to formulations, delivery systems, and / or methods of use having novel sustained release multi-phasic concentration patterns that can be used, among other things, to reduce mammographic breast density and / or breast stiffness in a warm-blooded animal. For example, the formulations can comprise: administering to a subject an effective amount of an androgenic agent and an effective amount of an aromatase inhibitor that provides a sustained release multi-phasic concentration pattern over time in the blood of the subject, as measured by serum concentration of the androgenic agent and plasma concentration of the aromatase inhibitor, and that improves, among other things, breast tissue stability and / or increases androgen receptor expression levels. The present disclosure also relates to the use of an effective amount of an androgenic agent in combination with an effective amount of an aromatase inhibitor for preventing or treating autoimmune inflammatory breast (AIM) in a patient in need thereof. Autoimmune inflammatory breast includes conditions of idiopathic inflammatory giant breast, plasma cell mastitis, granulomatous mastitis.

Description

Technical Field

[0001] This disclosure generally relates to pharmaceutical formulations and / or drug delivery systems having novel multiphase release modes when delivered to warm-blooded animals. The disclosed compositions can be used in warm-blooded animals where it is desirable to increase organ-specific or serum androgen levels without increasing or significantly increasing organ-specific and / or serum estradiol levels. This disclosure also relates to implants or transdermal patches containing effective amounts of an androgen agent and an aromatase inhibitor for delivery of multiphase release modes or pharmacokinetics, which can be used in warm-blooded animals where it is desirable 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 related to tissue-specific autoimmune inflammatory conditions.

[0002] Related References

[0003] Boyd et al., “Evidence linking breast tissue firmness to breast cancer risk”, PLoSOne, 2014 Jul 10; 9(7): e100937

[0004] Cheng Q, Jabbari K, Winkelmaier G, et al. Overexpression of CD36 in breast fibroblasts inhibited colony growth of breast cancer cell lines. Biochem Biophys ResCommun 2020; 526(1): 41-47.

[0005] Das L, Rai A, Vaiphei K, et al. Idiopathic gigantomastia: New mechanistic insights involving the paracrine environment. Endocrine 2019; 66(2): 166-177.

[0006] Dawson CA, Pal B, Vaillant F, et al. investigated the role of resident ductal macrophages in mammary epithelium and their role in promoting tissue remodeling. Nature Cell Biology, 2020; 22(5): 546-558.

[0007] DeFilippis RA, Fordyce C, Patten K, et al. Pressure signal transduction from human mammary epithelial cells contributes to the phenotype of mammographic density. Cancer Research 2014; 74(18): 5032-44.

[0008] 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.

[0009] Goulabchand R, Hafidi A, Van de Perre P, et al. Mastitis in Autoimmune Diseases: Literature Review, Diagnostic Approaches and Key Players in Pathophysiology. Journal of Clinical Medicine (J Clin Med) 2020; 9(4).

[0010] Gubbels Bupp MR, Jorgensen TN. Androgen-induced immunosuppression. Frontiers in Immunology (2018); 9: 794.

[0011] Guhl S, Artuc M, Zuberbier T, et al., reported that testosterone exerts selective anti-inflammatory effects on human skin mast cells in a cell subset-dependent manner. *Exp Dermatology*, 2012; 21(11): 878-80.

[0012] Jeremy Bercoff, “Shear Wave Elastic Imaging - White Paper”, SuperSonic Imagine, SA, copyright 2008.

[0013] 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*. *International Journal of Immunopharmacol*, 2020; 81: 106025.

[0014] Touraine P, Youssef N, Alyanakian MA, et al. Inflammatory macromastia in the context of immune-mediated diseases. Journal of Clinical Endocrinology and Metabolism, 2005; 90(9): 5287-94.

[0015] Uray IP, Liang Y, Hyder SM. Estradiol downregulates CD36 expression in human breast cancer cells. Cancer Lett 2004; 207(1): 101-7.

[0016] Walecki M, Eisel F, Klug J, et al. Androgen receptor regulation of Foxp3 expression in CD4+CD25+Foxp3+ regulatory T cells. Molecular Biology of the Cell, 2015; 10.1091 / mbc.E14-08-1323.

[0017] Wang H, Franco F, Tsui YC, et al. CD36-mediated metabolic adaptation supports the survival and function of regulatory T cells in tumors. Nature Immunology 2020; 21(3): 298-308.

[0018] U.S. Provisional Patent Application No. 62 / 067,297, filed on October 22, 2014, is entitled “Method for reducing breast density and / or the risk of breast cancer on mammography.”

[0019] U.S. Provisional Patent Application No. 62 / 324,525, filed on April 19, 2016, is entitled "Method for reducing breast density and / or the risk of breast cancer on mammography".

[0020] U.S. Patent No. 14 / 920,192, filed on October 22, 2015, is entitled “Method for reducing breast density and / or the risk of breast cancer on mammography.”

[0021] U.S. Patent No. 15 / 490,309, filed on April 18, 2017, is entitled “Method for reducing breast density and / or the risk of breast cancer on mammography.”

[0022] PCT / AU2015 / 000633, submitted on October 22, 2015, is titled "Methods for reducing breast density and / or the risk of breast cancer on mammography".

[0023] These references and each of PCT / AU2015 / 000633, U.S. Patent Nos. 62 / 067,297, 62 / 324,525, 15 / 490,309 and 14 / 920,192 are incorporated herein by reference in their entirety. Background Technology

[0024] Testosterone is crucial for regulating immune function in both men and women (Gubbels Bupp, 2018). Several physiological and disease conditions exist where it is desirable to increase tissue levels of 5α-dihydrotestosterone (DHT) while simultaneously decreasing estradiol levels within the same tissue without significantly perturbing serum estradiol levels. DHT is primarily a tissue-derived hormone of testosterone, transported to the tissue via circulation. Because the therapeutic window for administering testosterone without inducing androgenic side effects is relatively small, delivering testosterone to tissues in women requiring treatment is problematic.

[0025] In most tissues, there are also five α-reductases that convert testosterone into DHT. Therefore, there is an unmet need in therapeutic cases where it is desired to increase DHT without increasing estradiol. This disclosure relates to providing pharmaceutical formulations that aim to inhibit aromatase and induce 5α-reductase to achieve an alteration in the DHT to estradiol ratio within such tissues.

[0026] In both men and women, subcutaneous testosterone has been used to provide zero-order kinetics for testosterone administration. However, testosterone alone is not advisable in many physiological and disease states due to high levels of aromatase in the tissues undergoing therapeutic interventions. One example of this is women with high mammographic breast density, in whom tissue aromatase levels are very high, preferentially converting testosterone to estradiol. It is estimated that 43% of women aged 40 to 75 in the United States have mammographic breast density (MBD) classified as high, according to the Mammography Reporting and Data System (MDDS). Scores of 3 and 4 (or c and d) indicate that high breast density is a significant risk factor for breast cancer development, according to the American Cancer Foundation. Traditionally, treatment interventions during the perimenopausal transition have been low-dose combination oral contraceptives or continuous estradiol and synthetic progestin delivery systems to protect the uterus from an increased risk of endometrial cancer and unintended uterine bleeding. This is inappropriate for women with high breast density and / or breast firmness, as these can lower already dangerous testosterone levels and increase breast density and / or firmness. However, these are current recommendations from the North American Menopause Association and the Australian Menopause Association.

[0027] Another problem in the existing technology 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 report that breast pain affects their quality of life, and 10% report experiencing breast pain for at least half their lives. Treatment options are limited; tamoxifen and aromatase inhibitors have been used off-label for this condition. However, tamoxifen is associated with significant side effects affecting patient adherence, and aromatase inhibitors are contraindicated as monotherapy in premenopausal women due to their potential to disrupt the hypothalamic-pituitary-ovarian axis. There is a need in the art for better treatments to reduce breast pain in women.

[0028] Breast tissue elasticity is considered a factor in breast cancer development. Increased elasticity of breast cells has been shown to lead to increased mechanotransduction in the cellular genome, which may result in greater malignant transformation. There is a need in the art to provide compositions of pharmaceutical formulations that reduce mechanotransduction in the cellular genome to decrease the risk of malignant transformation.

[0029] A further problem in this field is tissue-specific autoimmune inflammatory conditions, particularly those occurring in the breast. These conditions are termed autoimmune inflammatory mastitis (AIM). The salivary glands and breast tissues share similar anatomical, histological, and physiological functions because they both fall at effector sites of the associated mucosal immune system, making them susceptible to autoimmune influences. Compared to normal breast tissue, autoimmune breast tissue has been shown to have elevated aromatase (which converts androgens to estradiol) and other inflammation-related factors, such as IGF2, EGFR, TGF-β, PDGFR-α, and β, which have been found to be upregulated. AIM is often treated with large doses of immunosuppressive therapy and / or disfiguring surgery, thus necessitating improved treatment approaches.

[0030] How a drug composition is delivered and its effects on the body may relate to the pharmacokinetic and / or pharmacodynamic profile of the drug composition, the drug delivery system, and / or the location of delivery to the subject's body. Broadly speaking, pharmacokinetics describes the rate and extent of a drug's absorption, distribution, and / or elimination from the body. In other words, how the body or a part of the body affects a drug composition. Broadly speaking, pharmacodynamics describes how a drug affects the body or a part of the body. Drug compositions may have different pharmacokinetic and / or pharmacodynamic profiles. For example, some drug compositions may have zero-order, first-order, or second-order kinetics. Kinetics and / or mechanics may affect the efficacy and / or therapeutic efficacy of a drug composition. Therefore, the role of pharmacokinetics and / or pharmacodynamics in certain drug compositions or drug delivery systems may be relevant to the outcome for a particular drug, drug composition, and / or subject.

[0031] Previously, implants combining testosterone with aromatase inhibitors have been produced, enabling the inhibition of testosterone's conversion to estradiol. However, if the manufacture and concentration of the compound result in the delivery of the active ingredient via zero-order kinetics, the end result is a decrease in serum estradiol levels, which is undesirable in many cases because it leads to estrogen deprivation symptoms and potential side effects from higher sustained levels of aromatase inhibitors. There is a continuing need for pharmaceutical formulations and / or delivery systems that can provide a high intratissue DHT to estradiol ratio. There is also a continuing need for alternative treatments to currently available therapies for diseases and conditions in which it is beneficial to increase tissue levels of 5α-dihydrotestosterone (DHT) while simultaneously decreasing intratissue estradiol levels without significantly perturbing serum estradiol levels.

[0032] This disclosure, in one or more embodiments, relates to addressing and / or improving one or more disadvantages of the prior art, or at least providing a usable pharmaceutical formulation or therapeutic delivery system, as will become apparent from the disclosure herein. This disclosure, in one or more embodiments, may also provide other advantages and / or improvements as discussed herein. Summary of the Invention

[0033] To achieve a high intratissue DHT to estradiol ratio, the inventors have recognized the urgent need for a delivery system that delivers testosterone and an aromatase inhibitor in a multiphase release mode. In one or more embodiments of the pharmaceutical formulation described herein, an early peak in serum testosterone can be provided, which is rapidly blocked by a high level of aromatase inhibitor, thereby allowing induction of the 5α-reductase conversion of testosterone to DHT. The formulation can then provide a rapid reduction in the aromatase inhibitor to ensure complete blockage of estradiol production and a symptomatic decrease in serum estradiol levels. In tissues with aromatase overexpression, this multiphase release mode allows for an axis shift towards elevated DHT to estradiol levels. For example, if zero-order kinetics of both components are used, the release of estradiol, which would lead to adverse effects on women and undesirable prolonged exposure to levels of aromatase inhibitor higher than required for a therapeutic response, will not occur.

[0034] In one aspect of the 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 multiphase concentration pattern in the subject's blood over time, such as that measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphase concentration pattern in the subject's serum or plasma includes:

[0035] In the first time period, the androgen had a first peak concentration (Tmax) in serum, and the aromatase inhibitor had an increased concentration in plasma but below its Tmax concentration in plasma; and

[0036] The second time period, in which the androgen initially had a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor had its Tmax concentration in plasma.

[0037] In another aspect of the 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 multiphase concentration pattern in the subject's blood over time, such as as measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphase concentration pattern in the subject's serum or plasma includes:

[0038] In the first time period, the androgen had a first peak concentration (Tmax) in serum and the aromatase inhibitor had an increased concentration in plasma but below its Tmax concentration in plasma.

[0039] The second time period, in which androgen agents initially have decreased serum concentration levels followed by increased serum concentration levels and aromatase inhibitors have their Tmax concentration in plasma.

[0040] In the third time period, the androgen had a second peak serum concentration below Tmax, and the aromatase inhibitor concentration in plasma gradually decreased, falling below the androgen concentration during the third time period; and

[0041] In the fourth time period, the serum concentration of androgens gradually decreased, and the plasma concentration of aromatase inhibitors gradually decreased, with the decrease levels of both being roughly the same.

[0042] In another aspect of the 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 a pellet; the pellet, when administered subcutaneously to a subject, provides a sustained-release multiphase concentration pattern in the subject's blood over time, such as that measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphase concentration pattern in the subject's serum or plasma includes:

[0043] In the first time period, the androgen had a first peak concentration (Tmax) in serum, and the aromatase inhibitor had an increased concentration in plasma but below its Tmax concentration in plasma; and

[0044] The second time period, in which the androgen initially had a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor had its Tmax concentration in plasma.

[0045] In another aspect of the 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 a pellet; the pellet, when administered subcutaneously to a subject, provides a sustained-release multiphase concentration pattern in the subject's blood over time, such as that measured by serum concentrations of the androgen agent and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphase concentration pattern in the subject's serum or plasma includes:

[0046] In the first time period, the androgen had a first peak concentration (Tmax) in serum and the aromatase inhibitor had an increased concentration in plasma but below its Tmax concentration in plasma.

[0047] The second time period, in which androgen agents initially have decreased serum concentration levels followed by increased serum concentration levels and aromatase inhibitors have their Tmax concentration in plasma.

[0048] In the third time period, the androgen had a second peak serum concentration below Tmax, and the aromatase inhibitor concentration in plasma gradually decreased, falling below the androgen concentration during the third time period; and

[0049] In the fourth time period, the serum concentration of androgens gradually decreased, and the plasma concentration of aromatase inhibitors gradually decreased, with the decrease levels of both being roughly the same.

[0050] In at least some embodiments, the pharmaceutical formulation comprises: 60 mg to 120 mg of testosterone or its ester, 2 mg to 6 mg of aromatase inhibitor, more preferably 4 mg to 6 mg of aromatase inhibitor, and stearic acid; the pharmaceutical formulation is compressed into pellets with a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm; the pellets, when administered subcutaneously to a subject, provide a sustained-release multiphase concentration pattern in the subject's blood over time, as measured by serum concentrations of testosterone or its ester and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphase concentration pattern includes:

[0051] In the first time period, testosterone or its esters have a first peak concentration (Tmax) in serum, and the concentration of aromatase inhibitors in plasma increases but remains below their Tmax concentration in plasma; and

[0052] The second time period, in which testosterone or its esters initially have decreased serum concentration levels followed by increased serum concentration levels, and aromatase inhibitors have their Tmax concentration in plasma.

[0053] In at least some embodiments, the pharmaceutical formulation comprises: 60 mg to 120 mg of testosterone or its ester, 2 mg to 6 mg of aromatase inhibitor, more preferably 4 mg to 6 mg of aromatase inhibitor, and stearic acid; the pharmaceutical formulation is compressed into pellets with a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm; the pellets, when administered subcutaneously to a subject, provide a sustained-release multiphase concentration pattern in the subject's blood over time, as measured by serum concentrations of testosterone or its ester and plasma concentrations of the aromatase inhibitor; and the sustained-release multiphase concentration pattern includes:

[0054] In the first time period, testosterone or its esters have a first peak concentration (Tmax) in serum and the concentration of aromatase inhibitors in plasma increases but is lower than their Tmax concentration in plasma;

[0055] The second time period, in which testosterone or its esters initially have decreased serum concentration levels followed by increased serum concentration levels, and aromatase inhibitors have their Tmax concentration in plasma.

[0056] In the third time period, testosterone or its esters had a second peak concentration in serum below Tmax, and the concentration of aromatase inhibitors in plasma gradually decreased, falling below the concentration of testosterone or its esters during the third time period; and

[0057] In the fourth time period, serum concentrations of testosterone or its esters gradually decreased, and plasma concentrations of aromatase inhibitors gradually decreased, with both decreasing at roughly the same rate.

[0058] In some exemplary embodiments, the aromatase inhibitor does not exhibit zero-order release in the pharmaceutical formulation during the first time period.

[0059] In some exemplary embodiments, the aromatase inhibitor exhibits primary release in the pharmaceutical formulation during a first time period.

[0060] In some exemplary embodiments, the aromatase inhibitor does not exhibit zero-order release in the pharmaceutical formulation during the second time period.

[0061] In some exemplary embodiments, during the second time period, the androgen agent (e.g., testosterone or its ester) in the pharmaceutical formulation does not exhibit zero-order release.

[0062] In some exemplary embodiments, the aromatase inhibitor does not exhibit zero-order release in the drug during the third time period.

[0063] In some exemplary embodiments, the androgen agent does not exhibit zero-order release in the pharmaceutical formulation during the third time period.

[0064] In some exemplary embodiments, the aromatase inhibitor exhibits primary release in the pharmaceutical formulation during the third time period.

[0065] In some exemplary embodiments, the androgen agent exhibits primary release in the pharmaceutical formulation during the third time period.

[0066] In some exemplary embodiments, the aromatase inhibitor does not exhibit zero-order release in the pharmaceutical formulation during the fourth time period.

[0067] In some exemplary embodiments, the androgen agent does not exhibit zero-order release in the pharmaceutical formulation during the fourth time period.

[0068] In some exemplary embodiments, the aromatase inhibitor exhibits primary release in the pharmaceutical formulation during the fourth time period.

[0069] In some exemplary embodiments, the androgen agent exhibits primary release in the pharmaceutical formulation during the fourth time period.

[0070] In some exemplary embodiments of the pharmaceutical formulation, the first time period ends immediately after the androgen reaches its first peak concentration (Tmax) in serum. In some exemplary embodiments of the pharmaceutical formulation, the first time period ends between 5 and 14 hours. In some exemplary embodiments of the pharmaceutical formulation, the first time period ends between 5.5 and 13 hours.

[0071] In some exemplary embodiments of the pharmaceutical formulation, the second time period ends immediately after the aromatase inhibitor has its Tmax. In some exemplary embodiments of the pharmaceutical formulation, the second time period ends between 23 hours and 80 hours.

[0072] Furthermore, some embodiments relate to a pharmaceutical formulation comprising: approximately 80 mg of testosterone or its ester, approximately 4 mg of anastrozole, and approximately 2 mg of stearic acid; said pharmaceutical formulation is compressed into pellets with a diameter of 4.4 mm to 4.6 mm and a length of 4 mm to 7 mm; said pellets, when administered subcutaneously to a subject, provide a sustained-release multiphase concentration pattern in the subject's blood over time, as measured by serum concentrations of testosterone or its ester and plasma concentrations of anastrozole; said sustained-release multiphase concentration pattern includes: a first time period in which testosterone or its ester has a first peak concentration (Tmax) in serum and anastrozole in plasma The concentration increases but remains below its Tmax concentration in plasma; a second time period occurs, wherein testosterone or its esters initially have a decreasing serum concentration level followed by an increasing serum concentration level, and anastrozole has its Tmax concentration in plasma; a third time period occurs, wherein testosterone or its esters have a second peak serum concentration below Tmax, and the concentration of anastrozole in plasma gradually decreases, falling below the concentration of testosterone or its esters during the third time period; and a fourth time period occurs, wherein the serum concentration level of testosterone or its esters gradually decreases, and the concentration of anastrozole in plasma gradually decreases, with the decreases in both being approximately equal. Methods of using the pharmaceutical formulation are also disclosed.

[0073] Further embodiments relate to a pharmaceutical formulation 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 a pellet; the pellet, when administered subcutaneously 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 includes: a first time period in which the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor does not have its Tmax concentration in plasma; a second time period in which the androgen agent has a decreasing serum concentration level followed by an increasing serum concentration level and the aromatase inhibitor has its Tmax concentration in plasma; a third time period in which the androgen agent has a second peak concentration in serum below its Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases, and decreases to a concentration below that of the androgen agent during the third time period; and a fourth time period in which the androgen agent has a decreasing serum concentration level and the concentration of the aromatase inhibitor in plasma decreases.

[0074] Further embodiments relate to a method for reducing breast density measured by mammography as AVBD and / or VBD% in patients with this need, comprising administering a subcutaneous pellet to the patient, said pellet containing: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0075] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing VBD% in patients with this need, including administering subcutaneous pellets to patients, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0076] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing AVBD in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0077] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing AVBD and VBD% in patients with this need, including administering subcutaneous pellets to patients, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0078] Further embodiments relate to the use of pharmaceutical formulations in the treatment of patients in need of breast density measured by mammography via AABD and / or ABD% in patients requiring this treatment, including administering subcutaneous pellets to the patient, said pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0079] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing ABD% in patients with this need, including administering subcutaneous pellets to patients, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0080] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing AABD in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0081] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing AABD and ABD% in patients with this need, including administering subcutaneous pellets to patients, the pellets comprising: i) an effective amount of an androgen 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 mammographic breast density in a patient with a mammographic breast density of 7.5% or higher, comprising administering a subcutaneous pellet to the patient, said pellet containing: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0083] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing VBD% in patients with breasts having 7.5% or higher VBD%, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0084] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing ABD% in patients with breasts having 7.5% or higher ABD%, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0085] Further embodiments relate to pharmaceutical formulations in the treatment of... Use in a method of mammography for breast density in patients with a breast score of 3 or 4 (or c or d) includes administering a subcutaneous pellet to the patient, said pellet containing: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0086] Further embodiments relate to pharmaceutical formulations that reduce the presence of... Use in a method of mammography for breast density in patients with a breast score of 3 or 4 (or c or d) includes administering a subcutaneous pellet to the patient, said pellet containing: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0087] Further embodiments relate to the use of pharmaceutical formulations in methods for inducing breast involution in patients with such needs, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0088] Further embodiments relate to the use of pharmaceutical formulations in methods of inducing net cell death exceeding proliferation in the breast of a patient in need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0089] Further embodiments relate to the use of pharmaceutical formulations in methods for inducing net extracellular matrix degradation in the breast of a patient in need of this, exceeding the development of extracellular matrix, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0090] Further embodiments relate to the use of pharmaceutical formulations in methods for reversing cell counts and mammographic breast density in the breasts of perimenopausal patients, including administering subcutaneous pellets to the patient, said pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0091] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing mammographic breast density and perimenopausal symptoms in patients with this need, including administering subcutaneous pellets to the patient, said pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0092] Further embodiments relate to the use of pharmaceutical formulations in methods for use in premenopausal and / or perimenopausal women to reduce the risk of breast cancer and substantially without causing disturbances to the hypothalamic-pituitary axis and / or other endocrine axes such as the adrenal glands and / or ovaries.

[0093] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing breast firmness in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0094] Further embodiments relate to the use of pharmaceutical formulations in methods of relieving breast pain in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0095] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing breast elasticity in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0096] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing mechanotransduction on the cellular genome to reduce the risk of malignant transformation in patients who require it, including administering subcutaneous pellets to a patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0097] Further embodiments relate to the use of pharmaceutical formulations in methods of increasing the ratio of fibroglandular tissue to adipose tissue in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0098] Further embodiments relate to the use of pharmaceutical formulations in methods of increasing CD36 in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0099] Further embodiments relate to the use of pharmaceutical formulations in methods of stabilizing and / or increasing androgen receptor expression levels in the breast tissue of patients in need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0100] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing and / or treating macromastia in patients with such needs, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0101] Further embodiments relate to the use of pharmaceutical formulations in methods of increasing GCDFP15 in patients with this need, including administering subcutaneous pellets to patients, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0102] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing breast pain associated with mammographic images taken in patients in need of this, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0103] Further embodiments relate to the use of pharmaceutical formulations in methods of increasing the sensitivity of a patient's mammogram, including administering subcutaneous pellets to the patient, said pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0104] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing ABD% and / or AABD in patients in need, including administering subcutaneous pellets to patients, said pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0105] Further embodiments relate to the use of pharmaceutical formulations in methods for reducing BPE in patient MRI images, including administering subcutaneous pellets to a patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0106] Further embodiments relate to the use of pharmaceutical formulations in methods of reducing the size and / or number of cysts in patients with this need, including administering subcutaneous pellets to the patient, the pellets comprising: i) an effective amount of an androgen agent; and ii) an effective amount of an aromatase inhibitor.

[0107] Furthermore, further embodiments relate to the prevention or treatment of autoimmune inflammatory mastitis in patients who require this treatment.

[0108] In particular, in another aspect of the invention, a method for preventing or treating autoimmune inflammatory mastitis in patients with such need is provided, comprising administering to the patient 1) an effective amount of an androgen agent and 2) an effective amount of an aromatase inhibitor.

[0109] In this embodiment, the androgen and aromatase inhibitor may be administered to the patient in the same pharmaceutical formulation or in separate formulations. Therefore, this disclosure extends to all combination therapies for patients with autoimmune inflammatory mastitis. Combination therapy refers to the simultaneous or sequential administration of the androgen and aromatase inhibitor via the same or different routes (as determined by a physician or caregiver), allowing the androgen and aromatase inhibitor to exert their respective therapeutic effects within overlapping therapeutic windows.

[0110] Typically, in at least some of the embodiments described herein for the treatment of autoimmune inflammatory mastitis, androgen agents and aromatase inhibitors are administered in the same or different sustained-release drug formulations.

[0111] In a particularly preferred embodiment, one or more sustained-release drug formulations are used to deliver androgen agents and aromatase inhibitors subcutaneously to a patient, for example, in one or more solid dosage forms such as one or more pellets.

[0112] Most typically, androgen agents and aromatase inhibitors are provided in the same sustained-release pharmaceutical formulation. The sustained-release formulation can be, for example, the pharmaceutical formulation described herein, designed to provide a sustained-release multiphasic concentration pattern in the blood of a subject over time, as measured by serum androgen concentrations and plasma and aromatase inhibitor concentrations.

[0113] Therefore, further embodiments relate to the use of the pharmaceutical preparation in a method of treating autoimmune inflammatory mastitis in patients with this need, including administering the pharmaceutical preparation to the patient in the form of a subcutaneous pellet containing: i) an effective amount of an androgen; and ii) an effective amount of an aromatase inhibitor.

[0114] The autoimmune inflammatory mastitis of patients treated according to this disclosure can be selected from, for example, the group consisting of: idiopathic inflammatory macromastitis, plasma cell mastitis, granulomatous mastitis, and combinations thereof.

[0115] Therefore, further embodiments of this disclosure relate to the use of a pharmaceutical preparation in a method of treating idiopathic inflammatory macromastia associated with an autoimmune disease in patients with this need, including administering the pharmaceutical preparation to a patient in the form of a subcutaneous pellet containing: i) an effective amount of an androgen; and ii) an effective amount of an aromatase inhibitor.

[0116] Further embodiments of this disclosure relate to the use of a pharmaceutical preparation in a method of treating plasma cell mastitis in a patient with this need, including administering the pharmaceutical preparation to the patient in the form of a subcutaneous pellet containing: i) an effective amount of an androgen; and ii) an effective amount of an aromatase inhibitor.

[0117] Further embodiments of this disclosure relate to the use of a pharmaceutical preparation in a method of treating granulomatous mastitis in a patient with such need, including administering the pharmaceutical preparation to the patient in the form of a subcutaneous pellet containing: i) an effective amount of an androgen; and ii) an effective amount of an aromatase inhibitor.

[0118] The invention is further extended to the use of a combination of an androgen agent and an effective amount of an aromatase inhibitor in the preparation of a medicament for the prevention or treatment of autoimmune inflammatory mastitis in patients with such need.

[0119] In yet another embodiment, the use of a combination of an aromatase inhibitor and an effective amount of an androgen agent in the preparation of a medicament for the prevention or treatment of autoimmune inflammatory mastitis in patients with such need is provided.

[0120] Typically, the androgen agent administered or contained in pharmaceutical formulations for the treatment of 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 their pharmaceutically acceptable salt or ester form.

[0121] As can be clearly seen from the foregoing, this disclosure is explicitly extended to methods for administering an effective amount of a pharmaceutical preparation as described herein to a subject for said purpose and / or treatment. Furthermore, in some embodiments, the use of androgens and aromatase inhibitors in the preparation of the pharmaceutical preparation as described herein is provided. The overview provided above is not intended to limit the embodiments disclosed herein or other embodiments disclosed in this specification. Moreover, the limitation of one embodiment may be combined with the limitation of other embodiments to form additional embodiments.

[0122] definition

[0123] Unless otherwise defined below, the terms used herein are as commonly used in the art:

[0124] The term "absolute area of ​​breast density" (AABD) refers to a measurement of the surface area of ​​fibroglandular tissue in a mammogram of a subject, expressed in square centimeters. This can be measured, for example, using the CUMULUS software algorithm or by visual examination of a mammogram. Several other tests can be used to measure AABD, including but not limited to VOLPARA, QUANTRA, and methods that take into account the surface area of ​​fibroglandular tissue in a mammogram.

[0125] The term "androgen agent" refers to an agent that increases androgen activity and / or synthesis. For example, an androgen agent can be a steroid hormone that binds with high affinity (in the pM or nM range) and specificity to its intracellular mediator androgen receptor, causing a conformational change in the receptor that allows for cofactor inclusion, nuclear transport, and / or stimulation of transactivation activity. This thus regulates the expression of target genes. For example, an androgen agent can be an androgen, such as androgens selected from the group consisting of testosterone, methyltestosterone, and dehydroepiandrosterone. Combinations of these androgen agents may also be considered. In addition to pharmaceutically acceptable esters of testosterone, esters may include, but are not limited to, heptaesters, propionates, cyclopentylpropionates, phenylacetic acid esters, acetate esters, isobutyrate, cyclobutyl ester, heptaester, decanoate, undecanoate, decyl ester, and / or isodecyl ester.

[0126] The term "area density percentage of the breast" (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 by visual examination of mammograms. Several other tests that can be used to measure ABD% include, but are not limited to, VOLPARA, QUANTRA, and methods that take into account the surface area of ​​fibroglandular tissue on mammograms.

[0127] 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, aromatase inhibitors can be selected from the group consisting of anastrozole, exemestane, and letrozole.

[0128] The term "autoimmune inflammatory mastitis (AIM)" refers to a non-infectious, non-lactational inflammation of the breast tissue caused by an autoimmune response to components within the breast tissue.

[0129] The term "breast cancer" refers to the malignant proliferation of epithelial cells lining the mammary ducts or lobules.

[0130] The term "breast elasticity" refers to a measurement of the pressure required to achieve deformation of a given portion of the breast or breast tissue. 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 could be directly measuring the elasticity of breast tissue by applying shear waves through the breast tissue, for example, using a method utilizing SuperSonic Imagine's ShearWave. TM SuperSonic Imagine Aixplorer by Elastography TM To measure.

[0131] The term “breast firmness” in its broadest sense refers to a measurement of the breast’s tolerance to deformation. Factors that may influence the degree of breast firmness include, but are not limited to, physical forces resulting from interactions between cells and between cells and the extracellular matrix, the number of cells present in the breast and the amount of collagen, the degree of fluid retention within the breast, the degree of proteoglycan expression, and / or combinations thereof. An example of measuring breast firmness involves using the formula force / deformation (dN / cm), where dN represents ten Newtons and cm represents centimeters, where deformation can be determined as the difference between the radius of the semicircle of the mammographic region and the radius of the volumetric hemisphere, and the pressure is recorded by a mammogram, such as a digital mammogram. For example, according to Boyd et al., deformation can be determined as the difference between R1 and R2, where R1 is the uncompressed radius and R2 is the compressed radius (Boyd et al., “Evidence of breast tissue firmness in relation to breast cancer risk”, PLoS One, 2014 Jul 10; 9(7): e100937).

[0132] The term "breast tissue" refers to the collection of epithelial cells, stromal cells, extracellular matrix, and / or migratory cells located inside or near the breast.

[0133] The term "effective amount" or "drug effective amount" for a reagent or compound refers to an amount sufficient to provide the desired therapeutic effect and is non-toxic, having an acceptable profile of non-toxicity and / or an acceptable profile of side effects. The required amount can vary among patients depending on factors such as the patient's age, general condition, severity of the condition being treated, the specific reagent or compound administered, and one or more combinations of these factors. Those skilled in the art can determine the generally appropriate "effective amount" for an individual case by referring to relevant textbooks and literature and / or using routine laboratory procedures.

[0134] 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 the total breast area (measured by 2D) or volume (measured by 3D). Mammographic breast density (MBD) is a qualitative or quantitative estimate of the amount of fibroglandular tissue (FGT) within the breast. It can be an absolute amount of FGT or a relative amount of FGT to the amount of non-FGT (primarily animal fat or adipose tissue). It can be calculated based on surface area (cm²). 2 (i.e., AABD) or volume (cm) 3The absolute amount of FGT is estimated by AVBD (i.e., absolute volumetric breast density). The relative amount of FGT can be an estimate of the surface area of ​​FGT relative to non-FGT (as a percentage (i.e., ABD%)) or an estimate of the volume of FGT relative to breast volume (i.e., VBD%). Another method can be to determine absolute volumetric breast density (i.e., AVBD), which can be a 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 mammography-to-tomometry (DBT), quantitative tissue imaging of palpable tissue (VTIQ), and combinations thereof. It can be determined by 2-D measurement and / or using Density classification qualitative evaluation of MBD, in In the density classification, 1 (or a) is the least dense, and 4 (or d) is the densest. MBD can also be qualitatively and / or quantitatively assessed by 3-D measurements and / or using breast volume measurements, such as determining volumetric breast density, which is the ratio of fibroglandular (dense) tissue in the breast 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 (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 indicate 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 tests that take into account the volume of fibroglandular tissue (cm³). 3 The method.

[0135] The terms "patient" or "subject" refer to animals, including human species, that can be treated with the compositions, methods, and kits disclosed herein. Unless a specific sex is specifically specified or the context clarifies a sex, the term "patient" is intended to refer to both males and females. The term "patient" can also refer to a female-to-male transsexual.

[0136] The term "perimenopause" or "menopausal transition" refers to the period surrounding menopause, during which a woman's body naturally transitions to permanent infertility (menopause). Women may begin perimenopause at different ages, and signs of progress towards menopause, such as irregular menstruation, may be noticeable in their 40s or even as early as their mid-30s. During perimenopause, estrogen levels may fluctuate unevenly, menstrual cycles may lengthen or shorten, and menstrual cycles may begin in which the ovaries do not release an egg (ovulation). Other menopausal-like symptoms may be experienced during perimenopause, including but not limited to hot flashes, sleep problems, and / or vaginal dryness.

[0137] The term “perimenopausal symptoms” should be understood to include, but is 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; decreased fertility; changes in sexual arousal or needs; 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, or combinations thereof.

[0138] The term "pharmaceutically acceptable" refers to those compounds, reagents, materials, compositions, excipients, and / or dosage forms that, within reasonable medical judgment, are suitable for contact with human and / or animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with reasonable benefits and / or risks.

[0139] The term "pill" refers to a solid dosage form implant containing an effective amount of androgen and an effective amount of aromatase inhibitor. Pills or implants may have different suitable shapes and sizes, such as spherical, cylindrical, rectangular, square, or combinations thereof. They may have corners or rounded edges. In some embodiments, the pill or implant may be compressible.

[0140] The term "pharmaceutical formulation" refers to a formulation containing an effective amount of androgen and an effective amount of aromatase inhibitor, which, when administered to a warm-blooded subject, provides a sustained, multiphasic concentration pattern of androgen release in the subject's blood over time, as measured by serum androgen concentration and plasma aromatase inhibitor concentration. There may be at least two, three, or four different time periods. These time periods vary depending on one or more of the following: the formulation, the delivery system, the drug concentration, and individual physiological changes during the completion or substantial completion of formulation absorption. Typically, the formulation is absorbed for no less than 3 months and no more than 5 months. In some embodiments, the formulation may be delivered via a transdermal patch. In some embodiments, the formulation may be delivered in a solid form, such as a subcutaneous pellet. In some embodiments, the formulation may be delivered in a solid form, such as a compressed subcutaneous pellet.

[0141] The term "postmenopausal women" should be understood to include not only older women who have gone through menopause, but also women whose ovaries have been removed or destroyed by other means or whose estrogen production has been suppressed for other reasons, such as those who have undergone long-term corticosteroid use, have Cushing's syndrome, or have gonadal dysgenesis.

[0142] The term "subject" refers to an animal, including the human species, that can be treated with the compositions, methods, and kits disclosed herein. Unless a specific sex is specifically specified or the context clarifies a sex, the term "subject" is intended to refer to both males and females. The term "subject" can also refer to a female-to-male transsexual.

[0143] As used herein, the term “treatment” or “therapy” includes preventative (e.g., prophylactic) treatment and / or palliative treatment, and as used herein, “treatment” refers to the act of providing preventative and / or palliative treatment.

[0144] The term "volume breast density percentage (VBD%)" refers to the proportion or percentage of the volume of fibroglandular (dense) tissue relative to the total volume of tissue in the breast. This can be achieved, 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. Attached Figure Description

[0145] To better understand this disclosure, and to more clearly illustrate how it can be implemented according to one or more embodiments of this disclosure, reference will now be made to the accompanying drawings by way of example. A sustained release concentration pattern over time in the blood of a subject is provided, as measured by serum concentrations of testosterone or its esters and plasma concentrations of aromatase inhibitors.

[0146] Figure 1 Data on serum androgen concentrations and plasma aromatase inhibitor concentrations over 84 days are shown, according to certain exemplary embodiments.

[0147] Figure 2 Showing Figure 1 The data was expanded to further illustrate the first 42 days.

[0148] Figure 3 Serum testosterone concentrations in 11 subjects up to 7 days after administration are shown according to certain exemplary embodiments.

[0149] Figure 4 Serum testosterone concentrations in 11 subjects up to 85 days after administration are shown according to certain exemplary embodiments.

[0150] Figure 5 The plasma concentrations of anastrozole in 11 subjects 336 hours after administration are shown, according to certain exemplary embodiments.

[0151] Figure 6 The plasma concentrations of anastrozole in 11 subjects after 1680 hours following administration are shown, according to certain exemplary embodiments.

[0152] Figure 7A-BThe absorption rate of anastrozole according to certain embodiments is explained.

[0153] Figure 8A -B indicates the plasma concentration of anastrozole after implantation according to certain embodiments.

[0154] Figure 9 The observation frequency after implantation in Example 3 is shown.

[0155] Figure 10 This illustrates a flowchart example of the model tested in Example 3.

[0156] Figure 11 The final structural model used in Example 3 is shown.

[0157] Figure 12A -B shows the group and individual predictions used in Example 3.

[0158] Figure 13 illustrates 11 individual plots of observed and predicted plasma concentrations of anastrozole according to certain embodiments.

[0159] Figure 14 The visual predictive test (VPC) of post-implantation anastrozole concentration used in Example 3 is shown.

[0160] Figure 15A -B shows the one-compartment and two-compartment models used in Example 3.

[0161] Figure 16 The predictions for patient 10 from several different input models used in Example 3 are shown.

[0162] Figure 17 The diagram illustrates the effects of the androgen testosterone and aromatase inhibitors (T+Ai) on the immunoreactivity of breast tissue.

[0163] Figure 18 Western blot analysis of CD36 protein in three explant samples of normal breast tissue obtained from three perimenopausal women at baseline and 24 hours after culture is presented.

[0164] Figure 19 A is a mammogram before (left) and after (right) treatment according to this disclosure, showing a significant reduction in breast size and density.

[0165] Figure 19 B is Figure 19 Patient A's Volpara Solution before (left) and after (right) treatment TM Breast volume and density measurements showed that breast volume decreased (58%), returning to pre-illness size, and fibroglandular tissue decreased (71%).

[0166] Figure 19 C is Figure 19 The patient's breast MRI images before treatment (left) and three consecutive years after treatment show reversal of extreme background parenchymal enhancement on MRI during the three consecutive years after treatment cessation.

[0167] Figure 20 A is a mammogram of another patient before (left) and after (right) treatment according to this disclosure, showing a significant reduction in breast size and density.

[0168] Figure 20 B is Figure 20 Patient A's Volpara Solution before (left) and after (right) treatment TM Breast volume and density measurements showed a decrease in breast volume (10%) back to pre-illness size and a decrease in fibroglandular volume (41%).

[0169] Figure 21 A shows mammograms of another patient before (left) and after (right) treatment according to this disclosure, which show a significant reduction in breast size and density.

[0170] Figure 21 B is Figure 21 Patient A's Volpara Solution before (left) and after (right) treatment TM Breast volume and density measurements showed that breast volume decreased (32%) back to pre-illness size and fibroglandular volume decreased (52%).

[0171] Figure 22 A is a magnetic resonance imaging (MRI) image of the breast of another patient before (left) and after (right) treatment according to this disclosure, showing complete reversal of the MRI's extreme background parenchymal enhancement.

[0172] Figure 22 B is Figure 22 Mammographic images of the breasts of patient B before (left) and after (right) treatment, showing a decrease in breast size and density.

[0173] Figure 22 C is Figure 22 Volpara Solution for Patient B before (left) and after (right) treatment. TM Breast volume and density measurements showed that breast volume decreased (23%) back to pre-illness size and fibroglandular volume decreased (36%).

[0174] Figure 23A shows extreme breast swelling in another patient, confirmed by mammography prior to treatment according to this disclosure.

[0175] Figure 23 B shows a magnetic resonance imaging (MRI) image of a breast with very little fat and strong metabolic activity, such as Figure 23 The extreme background parenchymal enhancement in the breast of patient A is shown.

[0176] Figure 24 A shows an MRI image of another patient with reactive axillary lymphadenopathy that developed granulomas in the retroareolar space.

[0177] Figure 24 B is Figure 24 MRI images of the breast of patient A, showing regression of granulomas and reduction in axillary lymph node size after treatment according to this disclosure.

[0178] In view of the following detailed description and the appended claims, those skilled in the art will understand that various substitutions and / or modifications can be made to this disclosure without departing from the scope of the claimed invention. Detailed Implementation

[0179] 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. Furthermore, individual features or combinations of features in certain embodiments may constitute additional embodiments. The specific structural and functional details disclosed herein should not be construed as limiting, but are merely representative of the methods for teaching those skilled in the art to use the disclosed embodiments and variations thereof in various ways.

[0180] The headings included in the detailed description are for the reader's convenience only and should not be used to limit the subject matter found throughout the disclosure or claims. These headings should not be used to interpret the scope or limit of the claims.

[0181] This disclosure relates at least in part to providing a drug delivery system having a multiphase sustained release pattern comprising an androgen agent and an aromatase inhibitor in a compressed pellet, unexpectedly overcoming the problem of achieving a high intratissue DHT to estradiol ratio when delivered subcutaneously to a subject. The desired effect is that the early peak of serum testosterone is rapidly blocked by a high level of the aromatase inhibitor, thereby inducing the conversion of testosterone to DHT via 5α-reductase. A rapid reduction of the aromatase inhibitor is then desired to ensure the absence or substantial blockage of overall estradiol production and a symptomatic decrease in serum estradiol levels. By utilizing certain exemplary embodiments exhibiting this multiphase sustained release pattern, a shift towards high DHT to estradiol levels can be achieved in tissues with aromatase overexpression.

[0182] Several instances exist where aromatase overexpression leads to disease processes, in which both women and men can benefit from this combination of testosterone and aromatase inhibitors. Therefore, in these tissues, this enzymatic combination, where aromatase overexpression and 5α-reductase co-expression are likely present, could be pharmacologically manipulated to achieve an ordered shift toward a change in the DHT / estradiol ratio. These disease conditions include (but are not excluded) hypermastographic breast density, breast pain, endometriosis, gynecomastia, perimenopausal and premenstrual tissue abnormalities, and autoimmune inflammatory mastitis (including conditions such as idiopathic inflammatory macromastitis, plasma cell mastitis, and granulomatous mastitis).

[0183] Figure 2 Examples of combinations of testosterone and aromatase inhibitors are illustrated, as well as the effects of this regimen on several subjects (N=11) who had received a subcutaneous pellet regimen.

[0184] In this example, Phase I showed a rapid increase in both serum testosterone and plasma anastrozole in the subject. This increase led to rapid tissue induction of 5α-reductase and severe inhibition of aromatase. The high tissue levels of testosterone were rapidly converted to DHT, and the conversion to estradiol was completely or substantially blocked. In this example, Phase I lasted approximately 8 days, as indicated by the vertical line separating Phase 1 from Phase 2. The duration of Phase 1 can vary due to a variety of factors, including but not limited to: the biology of the specific subject, the formulation of the pellet, and / or the manufacturing parameters used to manufacture the pellet.

[0185] In this example, Phase II lasts approximately 14 days. In Phase 2, the enzymatic system is readjusted to induce sustained high levels of anastrozole in tissues, which continues to allow the induction of 5α-reductase to function unopposed or substantially unopposed for the aromatase conversion of testosterone and estradiol. This readjustment results in a decrease in serum testosterone, avoiding excessive androgenic side effects and sustained 5α-reductase induction. In this example, Phase II lasts approximately 14 days, as indicated by the vertical line separating Phase 2 from Phase 3. The duration of Phase II can vary due to a variety of factors, including but not limited to: the biology of the specific subject, the formulation of the pellet, and / or the manufacturing parameters used to manufacture the pellet.

[0186] In this example, the anastrozole level in phase III rapidly decreases, allowing some testosterone to be converted to estradiol and avoiding the exhaustion symptoms that can result from high levels of anastrozole observed in zero-order release mode. During this period, induced 5α-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 testicles are depleted. As in this example... Figure 1 As shown, the subjects were followed up for another 70 days. The duration of Phase 3 can vary due to a variety of factors, including but not limited to: the biology of the specific subject, the formulation of the pellets, and / or the manufacturing parameters used to manufacture the pellets.

[0187] Some embodiments can be used to treat extreme mammographic breast density. Serum reproductive hormone levels are weakly correlated with intratissue levels due to the large amount of inactive or active metabolites (such as testosterone) metabolized to prohormones. In certain disease states, such as extreme mammographic breast density, there is overexpression of enzymes known to convert testosterone to estrogen or dihydrotestosterone (which is 10 times more potent as an androgen than testosterone). By altering the androgen-to-estrogen ratio in high mammographic breast density, it is possible to reduce the density and thus reduce the incidence of breast cancer. The reduction in density also applies to conditions such as gynecomastia where there is an excess of estrogen / androgen in the tissue. To facilitate the alteration of the androgen-to-estrogen ratio, a subcutaneous pellet has been designed to promote a multiphasic sustained release pattern. When anastrozole and testosterone are combined in a specific manner and compressed into a specifically compressed pellet, the resulting pharmacokinetics promote this multiphasic sustained release of the two active ingredients. Similarly, as discussed elsewhere, it is desirable that an early increase in anastrozole, delivered to tissues expressing high levels of aromatase at approximately peak testosterone levels, achieves the minimum concentration required to promote or substantially block aromatase activity within those tissues. This early increase in anastrozole promotes the conversion of testosterone to dihydrotestosterone via 5α-reduction and fosters an environment of androgen excess relative to estrogen. Once this initial high level of anastrozole is achieved, it is maintained at a significantly low level to avoid inducing hypothalamic-pituitary changes or systemic aromatase inhibition, as seen in higher concentrations of aromatase inhibitors, for example, when taken orally. This then allows testosterone to slowly enter the tissues, where 5α-reductase is induced in the somatic system to continue perturbing the antigenic environment. This unique multiphasic sustained release pattern can be used to alter tissue hormone levels without causing significant physiological changes in other serum reproductive hormone levels and the resulting potential adverse side effects.

[0188] Certain embodiments may be used to treat one or more of the following: high breast density and breast stiffness. High breast density and / or breast stiffness are not normal but pathological; and sometimes can be addressed with effective amounts of androgens and / or aromatase inhibitors rather than with lifestyle modifications (e.g., diet and exercise), which have not been proven successful in premenopausal, perimenopausal, and / or postmenopausal women.

[0189] Some embodiments relate to pharmaceutical formulations that can be used to provide one or more individualized reductions in the following: mammographic breast density and breast firmness in a patient requiring this, including (i) determining the patient's MBD and / or breast firmness; (ii) optionally, measuring the patient's free androgen index and / or changes in the patient's free androgen index over a period of at least one month; (iii) determining an adjusted dose of androgen and 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 some embodiments, measuring the patient's free androgen index and / or changes in the patient's free androgen index over a period of at least one month may include collecting blood samples and measuring the amount of free androgen (or testosterone) in the patient's serum.

[0190] Some embodiments relate to pharmaceutical formulations that can be used to provide one or more of the following individualized reductions in VBD%, AVBD, and breast firmness in patients who require this, including (i) determining the patient's VBD% and / or AVBD and / or breast firmness; (ii) optionally, measuring the patient's free androgen index and / or changes 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 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 some embodiments, measuring the patient's free androgen index and / or changes in the patient's free androgen index over a period of at least one month may include collecting blood samples and measuring the amount of free androgen agent (or testosterone) in the patient's serum.

[0191] Some embodiments relate to pharmaceutical formulations that can be used to provide one or more of the following individualized reductions in ABD%, AABD, and breast firmness in patients who require this, including (i) determining the patient's VBD% and / or AVBD and / or breast firmness; (ii) optionally, measuring the patient's free androgen index and / or changes 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 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 some embodiments, measuring the patient's free androgen index and / or changes in the patient's free androgen index over a period of at least one month may include collecting blood samples and measuring the amount of free androgen agent (or testosterone) in the patient's serum.

[0192] Some embodiments involve pharmaceutical preparations that can be used to reduce breast density and breast firmness in mammograms of patients in need, wherein the pharmaceutical preparation increases the sensitivity of breast imaging detection by mammography, digital mammography, magnetic resonance imaging (MRI), ultrasound, digital mammography-to-the-tissue (DBT) combination, quantitative palpation tissue imaging (VTIQ), or a combination thereof.

[0193] Some embodiments involve pharmaceutical preparations that can be used to reduce breast density and breast firmness in mammograms of patients in need of such preparations, wherein the pharmaceutical preparation increases the detection of breast cancer development in the patient.

[0194] Some embodiments involve pharmaceutical preparations that can be used to reduce one or more of the following in patients who require it: VBD%, AVBD, and breast firmness, wherein the pharmaceutical preparation increases the detection of breast cancer development in the patient.

[0195] Some embodiments involve pharmaceutical preparations that can be used to reduce one or more of the following: ABD%, AABD, and breast firmness in patients who require this, wherein the pharmaceutical preparation increases the detection of breast cancer development in the patient.

[0196] The pharmaceutical preparations disclosed herein can be used to affect one or more of the following in patients: reduce mammographic breast density; treat mammographic breast density; reduce breast firmness; treat breast stiffness; reduce mammographic breast density in patients with a mammographic breast density of 7.5% or higher; reduce breast firmness in patients with breasts with a mammographic breast density of 7.5% or higher; reduce breast firmness in patients with breasts with a mammographic breast density of 7.5% or higher; reduce breast firmness in patients with breasts with breast firmness ... Mammographic breast density in patients with breasts rated 3 or 4 (or C or D); induction of breast involution in patients; induction of net cell death exceeding proliferation in patients' breasts; induction of net extracellular matrix degradation exceeding extracellular matrix development in patients' breasts; methods to reverse cell count and mammographic breast density in perimenopausal patients; reduction of mammographic breast density and perimenopausal symptoms in patients; and prevention or treatment of autoimmune inflammatory mastitis (e.g., idiopathic inflammatory macromastitis, plasma cell mastitis, granulomatous mastitis, and combinations thereof).

[0197] These pharmaceutical formulations and combinations of androgen agents and aromatase inhibitors (Ai) as described herein may be used in premenopausal, perimenopausal, and / or postmenopausal women.

[0198] For example, high breast density in perimenopausal women is known to be a risk factor for breast cancer development. Dense tissue in 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 receiving effective amounts of androgens such as testosterone and effective amounts of aromatase inhibitors (such as anastrozole) via a multiphase formulation may exhibit decreased breast density and / or breast firmness. The inventors have also found that premenopausal, perimenopausal, and / or postmenopausal women receiving effective amounts of testosterone and effective amounts of aromatase inhibitors (such as anastrozole) via a multiphase formulation may exhibit induced breast involution and / or net cell death exceeding proliferation. The inventors have further found that the effective amount of aromatase inhibitor delivered to the subject via breast tissue administered to the subject via a multiphase formulation can be used to prevent the conversion of testosterone to estrogen, thus allowing testosterone to induce breast cell involution.

[0199] Some further embodiments relate to the prevention or treatment of autoimmune inflammatory mastitis (AIM), including conditions such as idiopathic inflammatory macromastitis, plasma cell mastitis, and granulomatous mastitis as described above.

[0200] Breast tissue can be a target tissue for autoimmune diseases, a process favored by the hormonal environment (Touraine, 2005). Compared to normal breast tissue, autoimmune breast tissue has been shown to have elevated aromatase (which converts androgens to estradiol) and other inflammation-related factors such as IGF2, EGFR, TGF-β, PDGFR-α, and β (Das, 2019).

[0201] The breast is unique in that it requires an immune-privileged lactational environment and rapid tissue remodeling after lactation (known as lactational involution), which necessitates immunosuppression to avoid autoimmunity (Dawson, 2020). At its core is the formation of regulatory T (Treg) cells, which are capable of suppressing the proliferation of effector T cells and the production of cytokines, and play a crucial role in immune responses and the prevention of autoimmune diseases. During T cell development, T cell receptor (TCR) gene fragments are rearranged to produce a diverse TCR repertoire necessary for generating immunity against 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 in the thymus promote self-tolerance (known as central tolerance): (1) negative selection, thereby eliminating self-reactive T cells; and (2) CD4 + Forkhead frame P3 (Foxp3) +The generation of regulatory T (Treg) cells. At the heart of this process is CD36-mediated transfer of cell surface antigens to promote tolerance to host antigens during homeostasis. The major regulator of Treg cell development and function is the transcription factor Foxp3. Testosterone treatment induces a strong increase in the Treg cell population both in vivo and in vitro (Walecki, 2015). High levels of Treg cells in breast tumors reduce the effectiveness of immunotherapy due to their immunosuppressive capacity. Targeting Treg cells leads to unacceptable autoimmune side effects, limiting the use of this approach. The effects of testosterone and aromatase inhibitors on the immunoreactivity of breast tissue are illustrated graphically. Figure 17 .

[0202] The manifestations of AIM are associated with dense matrix replacement of breast adipose tissue, which is clearly shown on breast MRI in those affected by the disease (Touraine, 2005). One of the key factors 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, spleen cells, and some skin microdermal endothelial cells. It has been shown 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 breast cancer model subtypes (Cheng, 2020); and c) CD36 is severely suppressed in non-malignant tissues of women at high risk of breast cancer development due to high mammographic density (HMD) (i.e., breast tissue containing low fat) (DeFilippis, 2014).

[0203] Thea Tlsty's group (DeFilipis, 2014) proposed the following cascade: i) Increased 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 subsequently induces the profibrotic-like phenotype observed in HMD tissues.

[0204] As described herein, CD36 expression in breast tissue can be induced by treating tissue with a combination of an effective amount of androgen and an effective amount of aromatase inhibitor, and this disclosure is explicitly extended to such methods and use in patients such as perimenopausal, menopausal or postmenopausal women.

[0205] This article describes a subcutaneous pellet for the prevention or treatment of autoimmune diseases or conditions, designed to promote a multiphasic sustained release pattern to alter the androgen-to-estrogen ratio. As described herein, anastrozole and testosterone can be combined and compressed into pellets to provide pharmacokinetics that promote this multiphasic sustained release of the two active ingredients. This release pattern ensures that early high levels of testosterone are associated with very high levels of aromatase inhibitors, thus only DHT is delivered to the inflamed breast tissue, and testosterone is not converted to estradiol (which is pro-inflammatory). In the second phase, after peak testosterone levels are reached, 5α-reductase induction from the first phase continues to ensure the maintenance of high DHT levels for anti-inflammatory effects, while a faster decline in anastrozole occurs. Sustained high levels of anastrozole may lead to adverse events due to excessive systemic estradiol decline, but this risk is mitigated by the rapid decrease in anastrozole delivery. A third delivery phase then allows for the slow release of testosterone into the tissue, where 5α-reductase is induced in the somatic cellular system to continue perturbing the antigenic environment. This unique multiphase sustained release pattern can alter tissue hormone levels with minimal physiological changes in other serum reproductive hormone levels and the resulting potential adverse side effects. The effects of the androgen testosterone and the aromatase inhibitor (anastrozole, as described herein) on the immunoreactivity of breast tissue are shown in... Figure 17 .

[0206] 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 beneficially reset homeostatic mechanisms and reduce the estradiol to androgen ratio in tissues. This may result in one or more of the following advantages:

[0207] A. Enhanced mammographic detection due to decreased breast density allows mammography to detect malignant tumors at an earlier and / or less invasive stage.

[0208] B. Reduces the risk of interstitial breast cancer, such as those that may occur between mammogram screening rounds. These cancers are common in breasts with high MBD (mammary ductal diameter).

[0209] C. Reduce breast firmness.

[0210] D. Reduces pain during breast compression during mammography.

[0211] E. The ability to achieve better mammographic compression, at least in part, due to reduced pain.

[0212] F. Because better mammographic compression is achieved and breast tissue is less dense, less energy is needed to expose the image on a mammogram, thus reducing radiation to the breast tissue. This lowers the risk of radiation-induced breast cancer.

[0213] G. The ability to achieve better patient compliance when performing regular mammograms.

[0214] H. The ability to treat patients without causing disturbances to the hypothalamic-pituitary axis and / or other endocrine axes.

[0215] I. Reduces breast pain in patients.

[0216] J. Reduces the elasticity of the patient's breasts.

[0217] K. Reduce mechanotransduction of the cellular genome to lower the risk of malignant transformation in patients.

[0218] L. Increases the ratio of fibrous glands and adipose tissue in patients.

[0219] M. The patient's CD36.

[0220] N. Stabilizes and / or increases the level of androgen receptor expression in the patient's breast tissue.

[0221] O. To treat patients with macromastia.

[0222] P. Increases patients' GCDFP15.

[0223] Q. Reduce BPE in the patient's MRI images.

[0224] R. Reduce the size and / or number of cysts in patients.

[0225] S. Reduces the risk of breast cancer.

[0226] T. Treatment of autoimmune inflammatory mastitis (AIM) includes conditions such as idiopathic inflammatory macromastia, plasma cell mastitis, and granulomatous mastitis.

[0227] Diagnostic physicians and doctors use multiple categories to characterize the type and / or degree of breast density in a patient's mammogram.

[0228] 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 mammography-to-the-tissue (DBT) combination, quantitative tactile tissue imaging (VTIQ), and combinations thereof. Physicians may also use other indicators, such as medical or family history (to explain the genetic predisposition to breast density) and / or qualitative assessments of MBD, such as… (For example, the 5th edition uses a classification of breast parenchyma composition: “a” (the breast is almost entirely fat), “b” (dispersed areas of fibroglandular density), “c” (the breast is unevenly dense and may obscure small lumps), and “d” (the breast is very dense and reduces 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.

[0229] Breast pain is an important health concern for women. Breast pain is also associated with an increased VBD% and / or AVBD, breast stiffness, breast cancer risk, or a combination thereof. Some practices involve the use of androgen agents and aromatase inhibitors to reduce breast pain in patients.

[0230] Some embodiments relate to the use of subcutaneous pellets containing effective amounts of anastrozole and testosterone. In an exemplary embodiment, a subject is provided with 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 compressed pellet formulation. In an exemplary embodiment, a subject is provided with 4-6 mg of anastrozole and 60-120 mg of testosterone in a compressed pellet formulation. In an exemplary embodiment, a subject is provided with approximately 4 mg of anastrozole and approximately 80 mg of testosterone in a compressed pellet formulation. Other androgen agents may also be used. The duration of treatment for administering subcutaneous pellets containing anastrozole and testosterone may vary between 2 to 4 weeks, 3 months to 3 years, 6 months to 2 years, 3 months to 5 years, 1 to 5 years, or 1 to 3 years. In some embodiments, the treatment duration 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 some embodiments, the treatment duration 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 treatment duration may be approximately 3 years. The treatment may be applied to one or more of the following: a decrease in ABD%; a decrease in AABD; a decrease in VBD%; a decrease in AVBD; a decrease in breast pain; a decrease in breast firmness; a decrease in breast elasticity; a decrease in macromastia; a decrease in breast cysts; improved sensitivity and reduced false positives in mammography diagnosis; an increased ratio between fibroglandular and adipose tissue; and stable and / or increased androgen receptor expression levels.

[0231] Some embodiments relate to the use of subcutaneous pellets containing effective amounts of letrozole and testosterone. In an exemplary embodiment, a patient is provided with a subcutaneous pellet containing 0.5-20 mg of letrozole (4,4′-((1H-1,2,4-triazol-1-methylene)benzonitrile) and 40-130 mg of testosterone for subcutaneous administration. In an exemplary embodiment, a patient is provided with a subcutaneous pellet containing approximately 10 mg of letrozole (4,4′-((1H-1,2,4-triazol-1-methylene)benzonitrile) and 40-130 mg of testosterone for subcutaneous administration. Other androgen agents may also be used. The duration of treatment with subcutaneous pellets containing letrozole and testosterone can vary between 2 to 4 weeks, 3 months to 3 years, 6 months to 2 years, 3 months to 5 years, 1 to 5 years, or 1 to 3 years. In some embodiments, the treatment duration 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 some embodiments, the treatment duration 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 treatment duration may be approximately 3 years. The treatment may be applied to one or more of the following: a decrease in ABD%; a decrease in AABD; a decrease in VBD%; a decrease in AVBD; a decrease in breast pain; a decrease in breast firmness; a decrease in breast elasticity; a decrease in macromastia; a decrease in breast cysts; improved sensitivity and reduced false positives in mammography diagnosis; an increased ratio between fibroglandular and adipose tissue; and stable and / or increased androgen receptor expression levels.

[0232] Some embodiments relate to the use of subcutaneous pellets containing an effective amount of exemestane and an effective amount of testosterone. In exemplary embodiments, a patient is provided with a subcutaneous pellet containing 10-75 mg of exemestane, 6-methyleneandrost-1,4-diene-3,17-dione, and 40-130 mg of testosterone for subcutaneous administration. Other androgen agents may also be used. The duration of treatment with exemestane and testosterone can vary between 2 to 4 weeks, 3 months to 3 years, 6 months to 2 years, 3 months to 5 years, 1 to 5 years, or 1 to 3 years. In some embodiments, the duration of treatment can be approximately 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 can 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 can be approximately 3 years. Treatment may be applied to one or more of the following: reduced ABD%, reduced AABD%, reduced VBD%, reduced AVBD, reduced breast pain, reduced breast firmness, reduced breast elasticity, reduced macromastia, reduced breast cysts, improved sensitivity and reduced false positives in mammography, increased ratio between fibroglandular and adipose tissue, and stable and / or increased androgen receptor expression levels.

[0233] As described herein, the preparation of esters involves the functionalization of any hydroxyl and / or carboxyl groups that may be present, as will be understood by those skilled in the art of medicinal chemistry and drug delivery. For example, to prepare testosterone esters, the 17-hydroxyl group of the testosterone molecule is typically reacted with a suitable organic acid under esterification conditions. Such conditions usually involve 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, the ester can be reconstituted into a free acid using conventional hydrogenolysis or hydrolysis procedures.

[0234] The effective amount of androgen can vary depending on the androgen. Furthermore, the effective daily amount of testosterone may also vary. In some cases, an effective amount of testosterone can be delivered via a subcutaneous implant. In some embodiments, the effective amount of testosterone can be 40 to 200 mg. For example, in some 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.

[0235] In some embodiments, an effective amount of testosterone may be delivered in the form of a subcutaneous implant, such as a subcutaneous pellet containing 40 to 200 mg of testosterone, for example 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.

[0236] The effective daily dose of methyltestosterone may vary. In an exemplary embodiment, the effective dose of methyltestosterone may be from 0.1 mg to 10 mg, for example, 0.5 mg to 9 mg, 2 mg to 8 mg, 3 mg to 7 mg, or 4 mg to 5 mg. For example, the effective dose of methyltestosterone may be 0.5 mg, 1.25 mg, or 2.5 mg.

[0237] The effective dose of androgens used in combination with aromatase inhibitors may be relatively lower than the standard dose because patients have low levels of sex hormone-binding globulin in their serum, which may be caused by the aromatase inhibitor.

[0238] Sex hormone-binding globulins bind to androgens (such as testosterone) and transport them throughout the body. Its production is regulated by multiple mechanisms, but one of the factors influencing its levels is the amount of estrogen in the serum: higher estrogen levels correspond to higher levels of sex hormone-binding globulins and lower levels of free androgens. Conversely, lower estrogen levels correspond to lower levels of sex hormone-binding globulins and higher levels of free androgens, meaning that androgens have higher bioavailability. Therefore, after menopause, as estrogen levels decline, sex hormone-binding globulin levels decrease, and free androgens such as testosterone increase. These free androgens have multiple functions because androgen receptors are expressed in all cells of the body.

[0239] In some embodiments, dose levels below the lower limit of the above-mentioned androgen dosage range may be sufficient, while in other cases, higher doses above the upper limit of the above-mentioned range may still be used without causing any harmful side effects.

[0240] In some embodiments, the aromatase inhibitor may be, for example, a steroid aromatase inhibitor, a non-steroid aromatase inhibitor, and / or its isomers. Steroid aromatase inhibitors developed to date are based on a fundamental androstenedione core with chemical substituents incorporated at different positions of the steroid. Examples of steroid aromatase inhibitors include, but are not limited to, exemestane. And formestane. Other examples include mechanism-based steroid aromatase inhibitors that mimic the substrate, where the enzyme converts the inhibitor to a reactive intermediate, leading to aromatase inactivation. In some embodiments, the aromatase inhibitor is exemestane. Non-steroidal aromatase inhibitors can be classified into three classes: aminoglutethimide-like molecules, imidazole / triazole derivatives, and flavonoid analogs. Examples of non-steroidal aromatase inhibitors include anastrozole, exemestane, or letrozole. In some embodiments, the aromatase inhibitor is anastrozole or letrozole. In some embodiments, the aromatase inhibitor is anastrozole.

[0241] Aromatase inhibitors typically include third-generation aromatase inhibitors, such as anastrozole. Isimmetan and letrozole These third-generation aromatase inhibitors have revolutionized treatment for patients with hormone-sensitive breast cancer. The action of these aromatase inhibitors is specific because they effectively eliminate estrogen in the serum, thereby reducing sex hormone-binding globulin and enabling a synergistic effect.

[0242] In some embodiments, the aromatase inhibitor may be selected from the group consisting of anastrozole, exemestane, or letrozole. In some embodiments, the aromatase inhibitor is anastrozole or letrozole. In some embodiments, the aromatase inhibitor is anastrozole.

[0243] In some embodiments, a method for determining whether a patient has Breasts rated 3 or 4 (or C or D); breasts with a mammographic density of 7.5% or higher; breasts with dense mammograms; breasts with breast tissue equal to or greater than fat; breasts with more breast tissue than fat; breast cancer or a combination thereof.

[0244] In some embodiments, the patient has or is diagnosed with A breast score (grade 1-4) is between 2 and 4, for example, 2 to 3, or 3 to 4. In some embodiments, the patient has or is diagnosed with Breasts rated 2 or higher, for example A rating of 3 or 4, or The rating is 4.

[0245] In some embodiments, the patient has or is diagnosed with A breast grade (ad) is rated from b to d, for example, from b to c, or from c to d. In some embodiments, the patient has or is diagnosed with... Breasts rated B or higher, for example The rating is C or D, or The rating is d.

[0246] In some embodiments, the patient has or is diagnosed with a breast with a mammographic breast density of 7.5% or higher, such as 10%, 15%, 20%, 30%, 50%, 70%, or 95% or higher.

[0247] In some embodiments, the patient has or is diagnosed with a breast with a VBD% of 7.5% or higher, for example, a VBD% of 10% or higher, 15% or higher, 20% or higher, 30% or higher, 50% or higher, 70% or higher, or 95% or higher.

[0248] In some embodiments, the patient has or is diagnosed with a breast with an ABD% of 7.5% or higher, for example, an ABD% of 10% or higher, 15% or higher, 20% or higher, 30% or higher, 50% or higher, 70% or higher, or 95% or higher.

[0249] In some embodiments, the patient has or is diagnosed with A breast with a mammogram score of 3 (or c) and a breast density of 7.5% or higher, for example, a breast density of 10%, 15%, 20%, 30%, 50%, 70%, or 95% or higher on mammogram. In some embodiments, the patient has or is diagnosed with... A breast with a rating of 4 (or d) and a breast with a mammographic breast density of 7.5% or higher, for example, a mammographic breast density of 10% or higher, 15% or higher, 20% or higher, 30% or higher, 50% or higher, or 95% or higher.

[0250] In some embodiments, the patient has or is diagnosed with a breast with a VBD% ranging from 1% to 100%, for example, 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%. 25% to 50%, 25% to 49%, 30% to 100%, 30% to 95%, 30% to 90%, 30% to 80%, 30% to 70%, 30% to 60%, 30% to 50%, 30% to 40%, 40% to 100%, 40% to 95%, 40% to 90%, 40% to 80%, 40% to 70%, 40% to 60%, 40% to 50%, 50% to 100%, 50% to 95%, 50% to 90%, 50% to 80%, 50% to 75%, 50% to 74%, 50% to 70%, 50% to 60%, 75% to 100%, 75% to 95%, or VBD% of 75% to 90%. In some embodiments, the patient has or is diagnosed with a breast with a VBD% in the range of 10% to 40%.

[0251] In some embodiments, the patient has or is diagnosed with a breast with an ABD% ranging from 1% to 100%, for example, 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%. 25% to 50%, 25% to 49%, 30% to 100%, 30% to 95%, 30% to 90%, 30% to 80%, 30% to 70%, 30% to 60%, 30% to 50%, 30% to 40%, 40% to 100%, 40% to 95%, 40% to 90%, 40% to 80%, 40% to 70%, 40% to 60%, 40% to 50%, 50% to 100%, 50% to 95%, 50% to 90%, 50% to 80%, 50% to 75%, 50% to 74%, 50% to 70%, 50% to 60%, 75% to 100%, 75% to 95%, or ABD% of 75% to 90%. In some embodiments, the patient has or is diagnosed with a breast with an ABD% in the range of 50% to 100%. In some embodiments, the patient has or is diagnosed with a breast that appears dense on mammograms, such as a breast with breast tissue that is substantially equal to or greater than fat.

[0252] In some embodiments, the patient is a perimenopausal or postmenopausal woman.

[0253] In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the number of patients between one or more annual interventional mammography examinations. Scoring. For example, pharmaceutical formulations can be used to differentiate patients between one or more annual interventional mammograms. The score is reduced by 1 or more points, for example, by 2, 3, or 4 points between one or more annual interventional mammograms. In some embodiments, the pharmaceutical preparation can be used to reduce the patient's score between one or more annual interventional mammograms. A score decreases or reduces by 1 point, for example, by 2, 3, or 4 points between one or more annual interventional mammograms. In some embodiments, the pharmaceutical preparation maintains or stabilizes the patient's condition between one or more annual interventional mammograms. score.

[0254] The interval between one or more annual interventional mammograms can be from 1 to 20 years, for example, 1 year, 1.5 years, 2 years, 3 years, 4 years, 5 years, 6 years, 10 years, 15 years, or 20 years.

[0255] In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the mammographic breast density of a patient's breast between one or more annual interventional mammograms. In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the VBD% and / or AVBD of a patient's breast between one or more annual interventional mammograms. In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the ABD% and / or AABD of a patient's breast between one or more annual interventional mammograms. For example, this pharmaceutical preparation can be used to reduce or decrease the mammographic breast density of a patient's breast by 1% to 99% between one or more annual interventional mammograms, for example, by 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%. For example, pharmaceutical preparations can be used to reduce or decrease the VBD% and / or AVBD of a patient's breast by 1% to 99% between one or more annual interventional mammograms, for example, by 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%. For example, pharmaceutical preparations can be used to reduce or decrease the ABD% and / or AABD of a patient's breast by 1% to 99% between one or more annual interventional mammograms, for example, by 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 mammograms.For example, the pharmaceutical preparation 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 mammograms, for example, by at least 5%, 10%, 20%, 30%, 40%, 50%, 75%, 85%, 95%, or 99% between one or more annual interventional mammograms. For example, the pharmaceutical preparation 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 mammograms, for example, by at least 5%, 10%, 20%, 30%, 40%, 50%, 75%, 85%, 95%, or 99% between one or more annual interventional mammograms. For example, a pharmaceutical formulation may 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 mammograms, such as by at least 5%, 10%, 20%, 30%, 40%, 50%, 75%, 85%, 95%, or 99% between one or more annual interventional mammograms. In some embodiments, the pharmaceutical formulation may be used to maintain or stabilize the mammographic breast density of a patient's breast between one or more annual interventional mammograms.

[0256] In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the breast density of a patient's breast by at least 2% over a 4-hour period, such as 5%, 10%, 20%, or 30%, over a period of 8 hours, 24 hours, 1 day, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or 1 year.

[0257] In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the VBD% and / or AVBD of a patient's breast by at least 2% over a 4-hour period, such as 5%, 10%, 20%, or 30%, over a period of 8 hours, 24 hours, 1 day, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or 1 year.

[0258] In some embodiments, the pharmaceutical formulation may be used to reduce or decrease the ABD% and / or AABD of a patient's breast by at least 2% over a 4-hour period, such as 5%, 10%, 20%, or 30%, over a period of 8 hours, 24 hours, 1 day, 3 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or 1 year.

[0259] In some embodiments, the pharmaceutical preparation can be used to mitigate or reduce the risk of breast cancer development in a patient. For example, in some embodiments, the pharmaceutical preparation can be used to mitigate or reduce the risk of breast cancer development in a patient between one or more annual interventional mammograms. In some embodiments, the pharmaceutical preparation can be used to mitigate or reduce the risk of breast cancer development in a patient between one or more annual interventional mammograms and to avoid, mitigate, reduce, or reverse one or more perimenopausal symptoms. For example, one or more perimenopausal symptoms that can be mitigated, reduced, or avoided may include, but are not limited to, irregular menstruation; hot flashes and sleep problems; mood changes; emotional fluctuations; irritability; depression; vaginal dryness; urinary tract infection or vaginal infection; urinary incontinence; decreased fertility; altered sexual arousal or need; 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 combinations thereof.

[0260] In some embodiments, the pharmaceutical formulation may be used to increase or improve a patient's fat-to-breast tissue ratio between one or more annual interventional mammograms. 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 mammograms; for instance, it increases or improves a treated 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, or from 2:3 to 19:1 between one or more annual interventional mammograms. Improve to 19:1, increase or improve from 2:1 to 19:1, increase or improve from 4:1 to 19:1, increase or improve from 6:1 to 19:1, increase or improve from 8:1 to 19:1, increase or improve from 10:1 to 19:1, increase or improve from 1:19 to 10:1, increase or improve from 1:10 to 10:1, increase or improve from 1:4 to 10:1, increase or improve from 1:2 to 10:1, increase or improve from 3:2 to 10:1, increase from 3:1 to 19:1. Increase or improve 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. Increase or improve 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.

[0261] In some embodiments, the pharmaceutical preparation can be used to increase or improve a patient's fat-to-breast tissue ratio from 1:19 to 19:1, for example, 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 period of 4 hours, 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.

[0262] In some embodiments, the pharmaceutical formulation may be used to increase the percentage of fat in a treated patient's breast between one or more annual interventional mammograms. For example, a pharmaceutical preparation may increase the percentage of fat in a treated patient's breast by 1% to 99% between one or more annual interventional mammograms, for example, 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%.

[0263] In some embodiments, the pharmaceutical formulation increases the percentage of fat in the treated patient's breast by at least 2%, such as at least 5%, at least 10%, at least 25%, at least 40%, at least 75%, at least 95%, or at least 99%, over a period of 4 hours, 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.

[0264] In some embodiments, the pharmaceutical formulation enhances, increases, or improves breast compression during mammographic visualization or examination of the breast between one or more annual interventional mammograms. For example, the pharmaceutical formulation enhances, increases, or improves breast compression during mammographic visualization or examination of the breast between one or more annual interventional mammograms by a range of 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 some embodiments, as a result of enhancing, increasing, or improving breast compression during mammographic visualization or examination of the breast, the pharmaceutical formulation further reduces or alleviates patient pain during breast compression. For example, as a result of enhancing, increasing, or improving mammographic visualization or detection of breast compression, pharmaceutical preparations may further reduce, decrease, or minimize patient pain during breast compression 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 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.

[0265] In some embodiments, the pharmaceutical formulation reduces or decreases patient pain during breast compression. For example, the pharmaceutical formulation reduces or decreases patient pain during breast compression 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%, as a result of reducing, decreasing, 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 mammograms. For example, the pharmaceutical preparation may further enhance, increase, or improve breast compression during one or more annual interventional mammography examinations by a range of 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%.

[0266] In some embodiments, the pharmaceutical formulation reduces or decreases patient pain based on a visual analog scale (VAS) during breast compression. For example, the pharmaceutical formulation reduces or decreases VAS-based patient pain during breast compression so that the patient does not experience significant pain at 50-100mm, 50-80mm, 50-70mm, 60-100mm, 70-100mm, 80-100mm, or 90-100mm during one or more annual interventional mammograms.

[0267] In some embodiments, the pharmaceutical formulation enhances or improves patient adherence to regular mammographic visualization or testing, such as adherence to mammographic visualization or testing every 6 months, every year, every 2 years, every 3 years, or every 5 years.

[0268] In some embodiments, the pharmaceutical formulation reduces or decreases the radiation exposure required to visualize or examine a patient's breast during one or more subsequent mammograms, such as one or more subsequent annual mammograms. For example, the pharmaceutical formulation reduces or decreases the radiation exposure required to visualize or examine a patient's breast 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%.

[0269] In some embodiments, the pharmaceutical formulation induces breast involution in patients, such as perimenopausal patients.

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

[0271] In some embodiments, the pharmaceutical formulation induces net cell death exceeding proliferation in the breast of a patient, such as a perimenopausal patient.

[0272] In some embodiments, the pharmaceutical formulation reverses the number of cells and mammographic breast density in a patient's breast, such as in a perimenopausal patient's breast.

[0273] In some embodiments, the pharmaceutical formulation reduces or decreases breast firmness in a patient's breast, such as in a perimenopausal patient. For example, the pharmaceutical formulation reduces or decreases breast firmness in a patient's breast by a range of 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% annually. For example, the pharmaceutical formulation reduces or decreases breast firmness in a patient's breast by at least 5% annually, such as at least 8%, at least 10%, at least 15%, at least 20%, or at least 30%. In some embodiments, the pharmaceutical formulation reduces or decreases breast firmness 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%, over a 4-hour period, such as 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.

[0274] In some embodiments, the pharmaceutical formulation enhances, increases, or improves mammographic visualization or detection of a patient's breast, such as the breast of a perimenopausal patient. For example, the pharmaceutical formulation enhances, increases, or improves mammographic visualization or detection of a patient's breast by a 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 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% between one or more annual interventional mammograms. In some embodiments, the pharmaceutical formulation enhances, increases, or improves the visualization or detection of a patient's breast on mammography by at least 5%, such as 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, such as 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.

[0275] In some embodiments, the pharmaceutical formulation reduces mammographic breast density and avoids inducing masculinizing androgenic side effects or a hyperandrogenic state. For example, masculinizing androgenic side effects may include male pattern baldness, hirsutism or increased hair growth in unwanted areas, a deeper voice, acne, or a combination thereof. In some embodiments, the pharmaceutical formulation reduces mammographic breast density and eliminates the induction of masculinizing androgenic side effects or a hyperandrogenic state. In some embodiments, the pharmaceutical formulation reduces mammographic breast density and minimizes the induction of masculinizing androgenic side effects or a hyperandrogenic state.

[0276] In some embodiments, the pharmaceutical formulation significantly improves or enhances a patient's physical function, such as in relation to the patient's central nervous system, libido, musculoskeletal system, cardiovascular system, risk of autoimmune disease, severity of symptoms associated with autoimmune disease, or a combination thereof. For example, in relation to the patient's central nervous system, the pharmaceutical formulation may reduce depression, anxiety, general cognitive impairment including memory, or reduce the risk of dementia and Parkinson's disease. For example, in relation to the patient's libido, the pharmaceutical formulation may significantly improve overall libido, including the speed of sexual arousal and the ability to achieve orgasm. For example, in relation to the patient's musculoskeletal system, the pharmaceutical formulation may reduce inflammation and degenerative arthritis, improve bone mineral density, or improve muscle strength. For example, in relation to the patient's cardiovascular system, the pharmaceutical formulation may reduce foamy macrophage deposition in the arterial walls, reduce atherosclerosis, increase high-density lipoprotein, leading to improved cholesterol or high-density lipoprotein ratios. For example, in relation to the patient's risk of autoimmune disease, the pharmaceutical formulation may significantly reduce or decrease the patient's risk of developing autoimmune diseases such as Sjögren's syndrome, lupus, and rheumatoid arthritis. For example, when related to the severity of symptoms associated with a patient's autoimmune disease, such as Sjögren's syndrome, lupus, and rheumatoid arthritis, the pharmaceutical formulation can significantly reduce or decrease the severity of symptoms associated with the treatment of the patient's autoimmune disease. In some embodiments, the pharmaceutical formulation significantly improves or enhances a patient's physical function, such as cognitive function; reduces degenerative CNS diseases, including dementia or Parkinson's disease; muscle strength; libido; vitality; reduces monoamine oxidase-induced anxiety and depression; or combinations thereof.

[0277] In some embodiments, the pharmaceutical formulation further provides one or more of the following: i) reducing breast density on mammogram; ii) increasing the involutional effect on the patient's breast without converting testosterone to estrogen; iii) significantly reducing, lowering, or reversing perimenopausal symptoms; or iv) significantly improving or enhancing the patient's physical function, including cognitive function; reducing symptoms associated with degenerative CNS diseases, including dementia or Parkinson's disease; muscle strength; libido; vitality; reducing monoamine oxidase-induced anxiety and depression; or a combination thereof. In some embodiments, the pharmaceutical formulation further provides one or more of the following: i) reducing breast density on mammogram; ii) increasing the involutional effect on end organs (including the breast) affected by hormones without converting testosterone to estrogen; iii) significantly reducing, lowering, or reversing perimenopausal symptoms associated with fluctuations in estrogen levels; or iv) significantly improving or enhancing the patient's physical function, including cognitive function; reducing symptoms associated with degenerative CNS diseases, including dementia or Parkinson's disease; muscle strength; libido; vitality; reducing monoamine oxidase-induced anxiety and depression; or a combination thereof.

[0278] In some embodiments, the patient has a high free androgen index in their breast, such as 30% or higher, within four hours of administration of the androgen and aromatase inhibitor. In some embodiments, the patient has a supraphysiological free androgen index in their breast within four hours of administration of the androgen and aromatase inhibitor.

[0279] In some embodiments, treatment with a pharmaceutical formulation or combination of androgen and aromatase inhibitor as described herein further includes: a) measuring the level of free androgen index and / or aromatase inhibitor in serum, the serum being isolated from a blood sample taken from a patient at least 1 month after treatment; b) determining a subsequent dose, including a subsequent effective amount of androgen and a subsequent effective amount of aromatase inhibitor; and c) administering the subsequent dose to the patient.

[0280] In some embodiments, treatment with a pharmaceutical formulation or combination of androgens and aromatase inhibitors as described herein further includes: a) measuring the level of free androgens and / or aromatase inhibitors in serum, the serum being isolated from a blood sample taken from a patient at least one month after treatment, including centrifuging the patient's blood sample to separate the serum; b) determining a subsequent dose, including a subsequent effective amount of androgens and an subsequent effective amount of aromatase inhibitors; and c) administering the subsequent dose to the patient.

[0281] In some embodiments, the serum level of free androgen index measured in treated patients after 1 month can be 10-25%, for example 10-20%, 10-15%, 15-25%, 15-20%, 12-18%, 8-15%, or 11-14%.

[0282] In some embodiments, the serum level of free androgen index measured in treated patients after 3 months can be 2-10%, for example 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%.

[0283] In some 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 80-95% or 100%. For example, the administered aromatase inhibitor may reduce the aromatization of testosterone to estrogen in the subcutaneous fat of the patient's breast, pelvis, buttocks, abdomen, or combinations thereof, for example, by 80-95% or 100%.

[0284] In some 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, by 80-95% or 100%. For example, the administered aromatase inhibitor can reduce the aromatization of testosterone to estrogens in the subcutaneous fat of the patient's breasts, pelvis, buttocks, abdomen, or combinations thereof, for example, by 80-95% or 100%.

[0285] Annual mammographic density screening can be performed using an appropriate mammographic algorithm to identify a decrease in breast density. This algorithm measures the percentage of fibroglandular tissue volume to total breast volume (MBD). When this is a function of the average of two breast densities, the goal is to achieve an MBD of less than 10%. The rate of decrease in breast density should be at least 2% per year. If 2% is not achieved in the first year, an annual boost factor of 10% can be introduced in both TD and AD (V1(N) above). If the increase in androgen index (AI) is less than 10%, this annual boost factor is typically introduced only on an annual basis.

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

[0287] In some embodiments, administration of a combination of 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 some embodiments, the increase in bioavailability of dihydrotestosterone is greater than 25%, greater than 35%, greater than 45%, or greater than 55%.

[0288] For example, androgen agents used in pharmaceutical formulations or combination therapies as described herein may be selected from the group consisting of free testosterone, methyltestosterone, and / or dehydroepiandrosterone. In some embodiments, the androgen agent may be testosterone undecanoate, such as about 40 mg of testosterone undecanoate. For example, aromatase inhibitors may be selected from the group consisting of anastrozole, exemestane, or letrozole. In some aspects, the aromatase inhibitor may be anastrozole, such as about 1 mg of anastrozole.

[0289] In some embodiments, the pharmaceutical formulation may include an administration of an androgen agent or an androgen agent / aromatase inhibitor complex comprising, for example, an aromatase inhibitor linked by an ester bond, wherein the complex is generated by methods in the art.

[0290] In a preferred embodiment, both androgen agents (e.g., testosterone, methyltestosterone, and / or dehydroepiandrosterone) and aromatase inhibitors (e.g., anastrozole, exemestane, or letrozole) are administered subcutaneously, for example as implants such as pellets.

[0291] In some embodiments, androgen agents (e.g., testosterone, methyltestosterone, and / or dehydroepiandrosterone) and aromatase inhibitors (e.g., anastrozole, exemestane, or letrozole) are administered subcutaneously, for example, as pellets. For example, testosterone and anastrozole can be administered subcutaneously.

[0292] The appropriate dosing regimen for androgen agents, aromatase inhibitors, or pharmaceutical preparations containing androgen agents and aromatase inhibitors, the amount of each dose administered, and the intervals between compound doses may depend on a variety of factors, such as the specific aromatase inhibitor and androgen agent used in combination, the type of pharmaceutical preparation used, the type of physiological condition being treated, the characteristics of the subject being treated (e.g., species, age, weight, sex, medical condition, eating / fasting), the route of administration, and the severity of the condition being treated, or combinations thereof. A physician or diagnostician with general skills can readily determine and prescribe an effective amount of androgen agent, aromatase inhibitor, or pharmaceutical preparation to prevent or treat a specific physiological condition.

[0293] The drug formulation or preparation to be administered may contain a certain amount of a compound or its pharmaceutically acceptable salt or ester, in an amount effective in treating the condition of the subject being treated. Because two different compounds may be used together in combination therapy, the potency of each compound and the interaction effects achieved by combining them are usually also considered. Consideration of these factors for determining the therapeutically effective or preventatively effective dose required to improve side effects is entirely within the scope of a general technical clinician.

[0294] Administering a drug formulation containing androgens and aromatase inhibitors or combinations thereof to a subject includes self-administration and administration to a subject by another person (e.g., a doctor, nurse, healthcare worker, friend, etc.).

[0295] In some embodiments, the pharmaceutical formulation may be formulated in a manner compatible with the desired results. The pharmaceutical formulation may be administered in a convenient manner. The following examples of formulations are merely illustrative and are not intended to limit the scope of this disclosure.

[0296] In some embodiments, the pharmaceutical formulation may be an implant or pellet prepared by direct compression. For example, the pellet formulation may incorporate diluents, binders, lubricants, disintegrants, and active ingredients. 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 polyvinylpyrrolidone. Polyethylene glycol, ethylcellulose, and waxes may also be used as binders. The pellet formulation may require a lubricant to prevent the pellet and punch from sticking to the mold. Lubricants may be selected from smooth solids such as talc, magnesium stearate and calcium stearate, stearic acid, and / or hydrogenated vegetable oils. Typically, the hardness of the implant or pellet ranges from about 6 kg / N to about 10 kg / N, preferably from about 7 kg / N to about 9 kg / N, most preferably about 8 kg / N, and all such dosage forms within this range are explicitly provided herein.

[0297] By compressing the formulation into a solid dosage form, such as an implant in the form of pellets for subcutaneous administration, a sustained-release multiphasic concentration pattern as described herein can be obtained. Given that release patterns of such androgen agents and aromatase inhibitors are known to be achievable, those skilled in the art, in light of the disclosure provided herein, will be able to use different components in formulations and provide such pharmaceutical formulations according to this disclosure.

[0298] In some embodiments, the implant or pellet may be inserted into the subcutaneous fat of the subject's pelvis, the subject's breast, the subject's buttocks, the subject's abdomen, or a combination thereof. In some embodiments, the implant or pellet may be inserted into the subcutaneous fat of the subject's lower abdominal wall. In some embodiments, the implant or pellet may be inserted into the subcutaneous fat of the subject's upper buttock region.

[0299] As will be understood, androgen agents and aromatase inhibitors may be provided in pharmaceutical formulations other than those for subcutaneous administration, or together in the same formulation or in separate formulations for combination therapy as described herein. Such formulations contain one or more suitable fillers, lubricants, binders, disintegrants and / or pharmaceutically acceptable carriers or excipients as described above, or pharmaceutical formulations for providing a desired route of administration and / or for obtaining a desired release pattern (e.g., sustained release multiphase release pattern) of one or more active ingredients.

[0300] In some embodiments, androgen agents, aromatase inhibitors, or pharmaceutical formulations comprising combinations thereof may be provided as articles, such as kits, containing the active ingredients therein or in suitable pharmaceutical formulations, 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 agent or aromatase inhibitor in a pharmaceutical formulation in a container, package, or dispenser, along with instructions for administration to human subjects. The instructions may also include indications of satisfactory clinical endpoints or any adverse events that may occur, or any other information required by the Food and Drug Administration for use in humans.

[0301] Instructions may be on “printed materials,” such as on paper or cardboard inside the kit, on a label affixed to the kit or packaging material, or on vials or tubes containing kit components. Instructions may also be contained on computer-readable media, such as disks (floppy disks or hard disks), optical CDs (e.g., CD or DVD-ROM / RAM), magnetic tapes, electronic storage media such as RAM and ROM), and mixtures of these, including magnetic / optical storage media.

[0302] While this disclosure has been described with reference to certain exemplary embodiments to provide a better understanding of the disclosure, it should be understood that various modifications may be made without departing from the principles disclosed herein. Therefore, the invention should be understood to include such modifications within its scope.

[0303] Example

[0304] Example 1: A single-dose trial to evaluate the pharmacokinetics of testosterone and anastrozole from a subcutaneous testosterone and anastrozole implant (80 mg / 4 mg (T+Ai)) in women.

[0305] Pharmacokinetic analyses were performed on subcutaneous testosterone 80g and anastrozole 4mg implants in 11 healthy volunteers with high mammographic breast density. This study evaluated the modalities of both anastrozole and testosterone and allowed for pharmacokinetic modeling of this exemplary implant to demonstrate modalities and end-organ pharmacodynamic responses known to exist with excess aromatase activity, i.e., end-organ pharmacodynamic responses 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.

[0306] Ultrasonic evaluation of pellet dissolution was performed to demonstrate how a comprehensive analysis of reproductive hormone input and dissolution can be conducted to assess the potential impact on hypothalamic-pituitary function.

[0307] Main objectives:

[0308] The pharmacokinetic characteristics of testosterone after a single dose of T+Ai were evaluated by subcutaneous implantation.

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

[0310] Secondary objectives

[0311] To explore the potential impact of shear wave ultrasound assessment on breast tissue elasticity.

[0312] Implant preparation: Testosterone-anastrozole pills

[0313] Pellets for subcutaneous insertion were prepared by compressing sterile, non-micronized testosterone, anastrozole, and stearic acid in an 80:4:2 ratio into a 5 mm diameter cylindrical mold. The pellets were transferred to sterile 5 ml amber vials lined with sterile cotton lint and sealed with rubber stoppers and rolled aluminum tear seals. The vials were sterilized by gamma radiation at 25 kGy. USP-grade non-micronized testosterone and anastrozole were obtained from Azelis (Brookvale, NSW). NF-grade stearic acid was obtained from Medisca (Mascot, NSW). The steps involved in producing these pellets are summarized below:

[0314] 1. Testosterone (non-micronized), anastrozole and stearic acid were mechanically mixed and titrated over 2 hours.

[0315] 2. Mix thoroughly before titrating.

[0316] 3. The pellets are compressed using a press, and the resulting pellets have a hardness of 8 kg / N.

[0317] 4. Store the pills in a 5ml amber glass vial, with a sterile cotton pad on top.

[0318] 5. Sterilize the stored pellets with gamma at 25KyG.

[0319] 6. Correctly label the small balls.

[0320] 7. Store in a cool, dry place (room temperature; 20-25℃, 66-78°F).

[0321] method

[0322] Premenopausal women with high breast density (MBD) on mammograms were selected for the study. Successful candidates traveled to Wellend Clinic South Australia for administration. Participants were given medication during the luteal phase of their menstrual cycle to avoid the significant hormonal fluctuations of the follicular phase. Following implantation, PK blood sampling was performed continuously for three months. In short, sampling was repeated on the day of administration, daily during the first week, weekly for the next four weeks, and every second week for the final eight weeks, for a total sampling period of three months. The trial concluded on the final day of evaluation, day 85.

[0323] To determine whether the selected pharmacokinetic (PK) sampling protocol provided an adequate description of the T+Ai serum / plasma concentration-time curve, testosterone, dihydrotestosterone, and anastrozole were analyzed in the samples 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 after administration), without adjusting the timing of the existing time points. The sampling duration was not prolonged.

[0324] In summary, blood samples were collected before administration (within one hour of administration) and at 1, 2, 4, 6, 8, and 12 hours after administration 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 sample collection.

[0325] A total of 11 subjects were administered the drug and included in the safety analysis set. The pharmacokinetic (PK) analysis set included 11 subjects. Women were aged 35 to 55 years with a body mass index (BMI) ranging from 20 to 30 kg / m². 2 The demographic data of the participants are shown in Table 1.

[0326] Table 1: Summary of Participant Demographics (Security Analysis Set)

[0327] Demographic parameters (units) Number of participants average value Standard deviation Median Minimum value Maximum value 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 (White / Asian) 1 Race (non-Hispanic or Latino) 11

[0328] PK Results

[0329] Table 2 summarizes the PK parameters of plasma anastrozole and serum testosterone, and shows them in the table below. Figures 1 to 6 .

[0330] Table 2: Summary of PK parameters of plasma anastrozole and serum testosterone

[0331]

[0332]

[0333] Regarding pharmacokinetics, the median T value of anastrozole in plasma is... max The mean C was 2 days after administration (range 1-3 days). max =5.30 ng / mL, and the mean terminal half-life was 14 days. After 12 weeks, most participants had concentrations below the limit of quantitation (0.1 ng / mL). The 14-day half-life observed from T+Ai was 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 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 antitumor effects. The IC50 of anastrozole in tissues overexpressing aromatase (such as the MCF7 breast cancer cell line and the JEG3 human choriocarcinoma cell line) was 3.62 and 5.66 nM, respectively. Therefore, C max =5.3 ng / mL (18.07 nM) will cover the initially higher C of T. max =4.1 ng / mL and anastrozole remained at a consistently low level as T levels decreased. Therefore, using 4 mg of anastrozole in pellets achieved the concentration required to inhibit the aromatase IC50 in the target tissue. In contrast, a dose of 2 mg was insufficient.

[0334] For serum testosterone, median T max The mean C was 8 hours after administration (range 6-12 hours). max =4.1 ng / mL, compared to a baseline mean of 0.2 ng / mL. A second lower peak appeared at 2–3 weeks. Participants' testosterone levels continued to increase at 12 weeks compared to baseline, with a mean concentration of 0.8 ng / mL. Serum dihydrotestosterone was unmeasurable in most samples; in participants with measurable maximum values, T... max This is 2-3 days after administration. This example shows that the pellets, after subcutaneous insertion, display a pattern.

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

[0336] This case study provides an analysis of the reduction in breast density (MBD) on mammograms in women treated with a combination of testosterone and anastrozole. The study was conducted at the Wellend Clinic (Burnside War Memorial Hospital, Adelaide, South Australia). The primary indication for the intervention was one or more of the following: perimenopausal hormonal dysfunction, high MBD, which is considered a factor in reducing the risk of breast cancer (BC).

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

[0338] In 89 patients (14%), reducing MBD / BC risk was the primary indication for treatment; in 334 patients (51%), there was adjunctive hormonal dysfunction; and in 177 patients (27%), there were two indications. No primary indication was provided in 52 patients.

[0339] Note that 90 patients had a history of BC (14% of patients had a non-absent response to their BC history). Note the BC type of 20 patients – 14 had in situ BC and 6 had invasive BC.

[0340] It was noted that 222 patients (34%) had concomitant estrogen-based medications, most of which were in the form of topical medications (n=192).

[0341] As of the data cutoff in July 2017, 365 patients (56%) were still receiving T+Ai therapy. One patient had no treatment 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 deciding to discontinue therapy (n=46), therapy costs (n=30), the subject feeling that T+Ai therapy was ineffective (n=28), and the primary care physician advising the patient that T+Ai therapy had been completed (n=25).

[0342] Measurement of efficacy

[0343] Summary statistics of continuous variable measurements (including %VBD and AVBD measurements of MBD) were obtained from mammograms. Since no mammograms were scheduled for specific time points, an access window system was applied to the data to provide specific data values ​​for those time points. The selected window is as follows:

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

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

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

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

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

[0349] Drug dosage and its relationship with %VBD response

[0350] To examine the effect of T+Ai intervention on MBD (measured by %VBD), a subset of patients who had mammograms before and after treatment initiation were considered. For this subset of patients (n=142), using In the PRO CMIXED mixed model analysis, the change in MBD from baseline was used as the outcome measure. This procedure allows individual patients to contribute more than one mammogram after initiating T+Ai treatment (repeated measures analysis) and examines the possibility of different covariance patterns between the data.

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

[0352] i. Number of days since the first implantation

[0353] ii. Baseline %VBD MBD measurement (closest to but no later than the first T+Ai implantation value)

[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 the time of first implantation (years)

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

[0358] vii. Radiation dose from a mammogram

[0359] viii. Compression pressure in mammograms

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

[0361] x. Use of medications accompanied by estrogen (yes or no)

[0362] xi. Interaction term between the number of days since the first implantation and the cumulative dose of testosterone layer.

[0363] The last interaction term listed above allows for different slope fittings for each testosterone dose layer to see if there are potential differences between the layers.

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

[0365] proc mixed data = mmg3;

[0366] class TESTO_GP SUBJIDC 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 1 yr′1 1 1 / diVisor=3at MMG_DAY=365cl;

[0374] lsmestimate TES TO_GP′Across all dose levels at 2 yr′1 1 1 / divisor=3at MMG_DAY=730cl;

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

[0376] format TESTO_GP testo.BCA_HIST_N E2_USED_N yesno.MMG_MACH_N machine;

[0377] run.

[0378] Each term in the model statement represents an item from the previous bulleted list. Random statements are used to describe repeated measurements performed within each patient over time. An unstructured covariance structure is used because... Compared to the Composite Symmetric (CS) and Autoregressive (AR1) options, it provides a slightly better fit. The LSMEANS and LSMESTIMATE statements are used to estimate the changes in MBD from baseline at 1, 2, and 3 years, segmented by testosterone dose levels. Table 3 shows the parameter estimates obtained from the model.

[0379] Statistically significant findings were noted regarding the number of days since the first T+Ai implantation and the interaction between the number of days since the first T+Ai implantation and the cumulative testosterone level. Specifically, patients with a cumulative testosterone dose exceeding 500 mg showed a greater decrease in MBD over time compared to patients with a dose <500 mg. The cumulative anastrozole dose also approached a significance level of 0.05 (p = 0.06), and estimates further indicated that higher cumulative anastrozole doses were associated with greater MBD reduction. Baseline %VBD was also statistically significant, with higher baseline scores correlated with greater observed changes (reduction in %VBD).

[0380] Table 3: SAS PROC MIXED model estimation of MBD variation from baseline

[0381]

[0382] The least-squares mean of the change from baseline in %VBD of MBD estimated by cumulative testosterone layer at 1, 2, and 3 years is presented in Table 4 below. The highlighted rows show the estimated change from baseline, which are statistically significant (p < 0.05). Least-squares mean analysis of the estimated change from baseline in %VBD at 1, 2, and 3 years.

[0383] Table 4: Least squares of MBD change from baseline estimated based on cumulative testosterone dose and time since first implantation

[0384] Cumulative testosterone dose group Since the first implantation Least squares mean estimation 95% CI Above 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 <700mg 1 year -1.1249 -2.4165 0.1668 0.0872 500 to <700mg 2 years -1.6916 -3.0057 -0.3775 0.0121 500 to <700mg 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] The efficacy of T+Ai measured by AVBD in reducing MBD

[0386] In addition to the analysis performed using %VBD MBD measurements, supplementary analyses were performed using the same model discussed above in this example, with the change in absolute volume breast density (AVBD) from baseline as the dependent variable. The only other change to the list of dependent variables used was replacing 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 in the 700 mg + cumulative testosterone group, at two and three years after the first T+Ai implantation. The estimated changes in AVBD from baseline at two and three years were -22 cm, respectively. 3 and -60cm 3 .

[0387] Table 5: Least squares of AVBD variation from baseline estimated based on cumulative testosterone dose and time since first implantation.

[0388] Cumulative testosterone dose group Since the first implantation Least squares mean estimation 95% CI Above 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 <700mg 1 year 0.2747 -12.2756 12.8251 0.9652 500 to <700mg 2 years -4.1993 -19.8209 11.4223 0.595 500 to <700mg 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 mammogram information obtained from 142 patients who underwent mammograms before and after the start of T+Ai treatment, both %VBD and AVBD showed statistically significant changes from baseline MBD measurements after T+Ai therapy intervention.

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

[0392] When MBD was measured by AVBD, the significance values ​​were those in the 700 mg + cumulative testosterone groups at 2 and 3 years after the first T+Ai implantation. The estimated changes in AVBD from baseline at 2 and 3 years were -22 cm. 3 and -60cm 3 .

[0393] In Example 2, most women received anastrozole at a dose of 2 mg per implant. This dose was chosen to try to keep the dose as low as possible to avoid side effects. Although this dosing was effective, Example 2 shows 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 larger changes in MBD from baseline (decreases from baseline). Surprisingly and unexpectedly, the PK and PD of anastrozole shown in Example 1 suggest that larger doses of anastrozole (4 mg per implant) can be used to achieve efficacy while also avoiding or minimizing the side effects associated with the aromatase inhibitors disclosed herein.

[0394] Example 3: Pharmacokinetic profile of anastrozole obtained using modeling methods

[0395] This example provides a modeling approach to analyze the pharmacokinetic profile of anastrozole to evaluate the optimal input function describing the absorption of the assay described in Example 1. Table 6 lists the sampling schedule for anastrozole and testosterone. At each time point specified in Table 6, at least 8 mL of blood was collected via venipuncture for serum testosterone / DHT and plasma anastrozole. Figure 7A-B The anastrozole absorption rate is illustrated. The data in these figures are expressed as mean + / - standard deviation. Figure 7A shows the Y-axis on a logarithmic scale. Figure 7B shows the Y-axis on a linear scale. This figure was generated using the pellet absorption data summarized in Table 2 of Example 1. The percentage change in average volume from baseline was calculated and plotted over a period of months. Figure 8A -B plots the plasma anastrozole concentration after implantation. Figure 8A In the image, the data on the X-axis is plotted in hours after the data was ported. Figure 8B In the diagram, the X-axis is on a logarithmic scale. Figure 8A In -B, the hollow circle represents the observed data, the black line represents the median of the PK curve, and the dark gray dashed line represents the lower limit of quantitation (0.1 ng / mL). Figure 9 The observation frequency after implantation is shown. Approximately 15% of the data were below the lower limit of quantitation (BLQ). BLQ concentrations were observed before administration or 1000 hours after implantation.

[0396] Table 6

[0397]

[0398] In this example, a population modeling approach is used to characterize the pharmacokinetics (PK) of anastrozole. One-compartment and two-compartment models are fitted to the data. Several absorption models, such as first-order and zero-order, mixed-order, and transfer-compartment models, are tested. Model selection is based on statistical criteria (objective function) and visual inspection of diagnostic plots. Figure 10 A flowchart of the test model is shown. Figure 11 The final structural model used in this example is shown. This example illustrates that the best model for fitting the data is a two-compartment treatment model with absorption described by two inputs. Between-subjects variability (BSV) is included in apparent clearance (CL / F), apparent volume of distribution in the central compartment (Vc / F), and first-order absorption rate constant (KA). A proportional error model is used to describe unexplained variability (RUV) in residues.

[0399] Table 7 below provides parameter estimates for anastrozole. The parameters were estimated with good accuracy (RSE < 30%), except for inter-subject variability terms on Vc / F and apparent intercompartmental clearance (Q / F); however, this is considered acceptable given the available data. The model estimate for CL / F is 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] Figure 12A -B shows the group and individual predictions. Figure 12A The group predictions are displayed. Figure 12B Individual forecasts are displayed. The light gray line represents the identity line, and the dark gray line shows the data trend (Loess smoothing). Because BLQ data was removed, the forecast is... Figure 12B The trend line shows a tail. Overall, the model predicted the data well with minimal bias. The population predicted concentration is typical of patients. The individual predicted concentration is a posterior Bayes / individualized prediction. Figure 13 (ID: 1-11) shows separate plots of observed and predicted anastrozole plasma concentrations. Data are plotted at calibration time. The black dashed line equals the population model prediction, the gray solid line equals the individual model prediction, and the circle equals the individual observation data. The final model was used to simulate 1000 repetitions of the dataset to validate the model's simulation properties. The 95% CI near the median and 10% CI were calculated based on the simulated data. th and 90 th Predict the interval. Then, take the median of the observed data and 10... th and 90 th Percentiles are superimposed on the simulated data to validate the model.

[0403] Figure 14 Visual predictive computation (VPC) of anastrozole concentrations after implantation is shown. The VPC shows that the observed data are well captured by the simulated data from the model. The X and Y axes are on a logarithmic scale. The black dashed line = 10 of the observed values. th and 90 thPercentiles: Black solid line = observed median, gray shaded area = 80% prediction interval, light gray dashed line = lower limit of quantification.

[0404] Figure 15A -B shows a one-compartment to two-compartment model. Figure 15A This indicates a single-room layout. Figure 15B This represents a two-room layout with two inputs. The distribution of the conditionally weighted residuals (CWRES) over time for a single-room model is shown. Figure 15A The error indicates a misspecification in the elimination phase. In contrast, the two-compartment model ( Figure 15B The data shows that CWRES is uniformly distributed over time.

[0405] Figure 16 The figures show predictions for patient 10 from several different input models, including: single-level input (model 9), simultaneous level 1 and level 0 input (model 10), single level 0 input (model 11), and dual level 1 input (model 151). It is clear from the figures that models other than dual level 1 input failed to correctly capture the early stages of absorption.

[0406] Summarize

[0407] This analysis shows that the anastrozole PK profile follows a two-compartment arrangement. The two-input model accurately describes the anastrozole absorption curve. Other input models failed to capture the absorption phase correctly.

[0408] Example 4: Breast tissue elasticity

[0409] This example focuses on the potential impact on breast tissue elasticity assessed by shear wave ultrasound in subjects participating in the trial described in Example 1. Breast elasticity was measured by shear wave ultrasound on days 1, 29, 57, and 85. Four measurements of glandular tissue elasticity and adipose tissue elasticity were performed for each breast. The calculated average of the eight measurements for each tissue type was used to summarize and analyze the breast elasticity values. A summary of breast elasticity at each time point and changes from baseline is provided in Table 8. Table 8 shows that breast tissue elasticity gradually decreased over time, with glandular tissue decreasing by 28% from baseline and adipose tissue decreasing by 32% from baseline.

[0410] Table 8

[0411]

[0412] Example 6: Expression of CD36

[0413] Breast tissue in women with autoimmune inflammatory mastitis (AIM) has high levels of aromatase and estradiol, resulting in severe suppression of Treg expression. Induction of CD36 in breast tissue of women with AIM may lead to tissue-specific immunosuppression rather than systemic immunosuppression. This example demonstrates that CD36 can be expressed in normal breast tissue through treatment with a combination of androgens and aromatase inhibitors as described herein.

[0414] Normal breast tissue was removed from three perimenopausal women during surgery. Tissue samples were transported to the laboratory on ice, with a maximum time of one hour between excision and tissue processing. Breast tissue samples were washed to remove excess blood in RPMI-free medium supplemented with 5 ml of 200 mM glutamine (SAFC biosciences, Kansas, USA), 5 ml of 100x antibiotic / antifungal agent (Sigma, St Louis, MO, USA), 10 μg / ml insulin (Sigma, St Louis, MO, USA), and 10 μg / ml hydrocortisone (Sigma, St Louis, MO, USA).

[0415] Representative sections of each tissue sample were immediately fixed overnight in 4% formalin-phosphate-buffered saline (PBS) at 4°C, followed by dehydration using an automated tissue processor (Sakura Tissue-Tek VIP, USA) and solid paraffin embedding. Hematoxylin and eosin (H&E) staining was used to evaluate histopathology. Remaining fresh tissue was cut into small pieces (~3x3x1mm) and triplicated in 1cm sections. 3 Gelatin sponges (Spongostan; Johnson & Johnson, Skipton, UK) were pre-soaked and then half-immersed in treatment medium containing 10% dextran-coated carbon-treated fetal bovine serum (DCC-FCS) (SAFC biosciences, Kansas, USA) in 24-well tissue culture plates (BD Biosciences, NJ, USA). Tissue samples were then cultured for 24 hours in a medium containing 0.1% ethanol, 5 nM testosterone, and 25 ng / ml anastrozole.

[0416] Whole-cell lysates from control adipose tissue were prepared by sonication at 48°C in lysis buffer (1% Triton X-100, 50 mM KCl, 25 mM HEPES, pH 7.8, 10 mg / ml leucopeptide, 20 mg / ml aprotinin, 125 mM dithiothreitol, and 1 mM benzyl sulfonyl fluoride) and analyzed on the same Western blot. 50 mg of total protein sample was mixed with 50 ml of sodium dodecyl sulfate (SDS)-mercaptoethanol sample buffer and boiled for 10 min. The proteins were then separated on a 7.5% SDS gel and transferred to a polyvinylidene fluoride membrane. The membrane was then blocked for 1 hour at room temperature with 5% skim milk in phosphate-buffered saline (PBS) containing 0.5% Tween-20. Immunoblotting was performed using anti-human CD36 antibody diluted in PBS and horseradish peroxidase-conjugated secondary antibody diluted in PBS (Jackson Immunoresearch), followed by chemiluminescence detection (Amersham Bioscience, Buckinghamshire, England). Band density was measured by densitometrics using ImageMaster VDS and Image Quant Analysis Software (Amersham Pharmacia Biotech, Hong Kong). Relative protein levels of CD36 and β-actin in the raw total protein lysate from breast products were obtained. CD36 protein expression was normalized to β-actin expression. Antibodies containing synthetic peptides derived from the human CD36 sequence were generated by immunizing the host, purified by peptide affinity chromatography, and confirmed using control peptides. Figure 17 The results of Western blot analysis of CD36 protein in three explant samples at baseline and 24 hours after culture are shown. Cell lysates were immunoblotted with an anti-CD36 antibody. The experiment was performed twice, and the results were similar. Data are expressed relative to actin. A two-tailed Student's t-test showed significance at p = 0.00757. The results are listed in Table 9 below.

[0417] Table 9: Measurement of Western blot band density

[0418] 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

[0419] Increased CD36 levels can lead to the conversion of fibroblasts into adipocytes (fat). Adipocytes (fat) are semi-fluid and therefore more elastic. This example demonstrates a significant increase in CD36 in normal breast tissue following treatment with a combination of the androgen testosterone and the aromatase inhibitor anastrozole. Increased CD36 expression is associated with decreased breast firmness. As described herein, certain embodiments of this disclosure relate to the use of a combination therapy of androgens and aromatase inhibitors to increase CD36 and reduce breast firmness.

[0420] The above demonstrates that the combination of testosterone and aromatase inhibitors is highly effective in inducing CD36 in normal breast tissue, as shown by the Western blot below, which indicates that CD36 is largely induced by treatment of normal breast tissue harvested during surgery and grown in explants.

[0421] Example 7: Idiopathic inflammatory gigantomastia

[0422] The following discussion pertains to the treatment of four patients with idiopathic inflammatory mastitis associated with mild to moderate macromastia, according to this disclosure.

[0423] Patient 1 : 42.03416971

[0424] Medical visit

[0425] A 38-year-old chiropractor experienced rapid breast enlargement—"doubling in size"—along with periareolar inflammation and persistent, severe pain for two months, unresponsive to over-the-counter analgesics and NSAIDs, impacting her ability to work.

[0426] Bra cup size before the onset of the disease

[0427] The patient's bra cup size is 32A, and she has been unable to wear a bra since the onset of the disease.

[0428] Past medical history

[0429] The patient had severe premenstrual anxiety disorder partially controlled by oral contraceptives. She had stopped taking oral contraceptives two years prior to consultation, had regular 28-day cycles, two pregnancies, and two live births. Severe postpartum arthritis—undiagnosed—resolved after one year, with no abnormal blood parameters noticed.

[0430] VAS pain scale before onset: 95mm

[0431] treat

[0432] As described in Example 1, subcutaneous implantation of 80 mg testosterone and 4 mg anastrozole pellets was continued for 3 months, and this was repeated for two additional 3-month periods.

[0433] result

[0434] i. VAS pain scale 10 mm at 4 weeks

[0435] ii. No inflammation was detected at 4 weeks.

[0436] iii. 71% reduction in fibroglandular tissue

[0437] iv. Breast volume decreased by 58%, returning to its pre-illness size.

[0438] v. Complete reversal of extreme background parenchymal enhancement on MRI, 3 years after discontinuation of continuous therapy.

[0439] vi. Without SAE

[0440] vii. BMI remained unchanged.

[0441] Mammograms of the patient's breasts before and after treatment, as shown below. Figure 19 As shown in Figure A, and the MRI images before and 3 years after treatment are as follows: Figure 19 As shown in Figure C. The results of breast volume and density measurements are as follows. Figure 19 As shown in B.

[0442] Patient 2: 42.09761311

[0443] Medical visit

[0444] A 42-year-old nurse experienced rapid breast enlargement—"very hard," with diffuse inflammation and persistent, severe pain for four months, unresponsive to over-the-counter painkillers and NSAIDs, impacting her work and family life—she couldn't even ride a horse. Her left breast was significantly larger than her right.

[0445] Bra cup size: before the onset of illness

[0446] The patient's bra cup size is 36C, and she has been unable to wear a bra since the onset of the disease. She wears a sports bra all day long and even sleeps in it.

[0447] Past medical history

[0448] Regular 28-day cycles, two pregnancies and two live births with severe preeclampsia, postpartum depression, including hospitalization, with no noticeable abnormalities in blood parameters.

[0449] VAS pain scale before onset: 100mm

[0450] treat

[0451] Subcutaneous implantation of 80 mg testosterone and 4 mg anastrozole pellets continued for 3 months, with two additional 3-month periods repeated.

[0452] result

[0453] i. VAS pain scale 15mm at 4 weeks

[0454] ii. No inflammation was detected at 4 weeks.

[0455] iii. 41% reduction in fibroglandular tissue

[0456] iv. Breast volume decreased by 10%, returning to its pre-illness size.

[0457] v. No SAE

[0458] vi. BMI remained unchanged.

[0459] vii. No breast augmentation surgery

[0460] Mammograms of the patient's breasts before and after treatment, as shown below. Figure 20 As shown in Figure A. Breast volume and density measurements are as follows: Figure 20 As shown in B.

[0461] Patient 3: 42.09761311

[0462] Medical visit

[0463] A 37-year-old lawyer experienced rapid breast enlargement—inflammation, particularly around the nipples—and persistent, severe pain for two months, unresponsive to over-the-counter painkillers and NSAIDs—affecting her work and family life.

[0464] Bra cup size

[0465] Before the onset of the disease, my bra size was 34C. After the onset of the disease, I was unable to wear a bra and wore a sports bra all day, even sleeping in one.

[0466] Past medical history

[0467] Regular 28-day cycles, no pregnancy, moderate endometriosis, no abnormal blood parameters noticed.

[0468] VAS pain scale before onset: 100mm

[0469] treat

[0470] As described in Example 1, subcutaneous implantation of 80 mg testosterone and 4 mg anastrozole pellets was continued for 3 months, and this was repeated for two other 3-month periods.

[0471] result

[0472] i. VAS pain scale at 4 weeks: 0 mm

[0473] ii. No inflammation was detected at 4 weeks.

[0474] iii. Fibroglandular tissue decreased by 52%.

[0475] iv. Breast volume decreased by 32%, returning to its pre-illness size.

[0476] v. No SAE

[0477] vi. BMI remained unchanged.

[0478] vii. No breast augmentation surgery

[0479] Mammograms of the patient's breasts before and after treatment, as shown below. Figure 21 As shown in Figure A. Breast volume and density measurements are as follows: Figure 21 As shown in B.

[0480] Patient 4: 42.83593371

[0481] Medical visit

[0482] A 41-year-old police officer suffered from rapidly enlarging breasts—"very hard"—diffuse inflammation, and persistent, severe pain for a month. The condition was unresponsive to over-the-counter painkillers and NSAIDs, impacting the patient's work and family life—and she was unable to wear a Kevlar protective kit.

[0483] Bra cup size

[0484] Before the onset of the disease, she was 36B. After the onset of the disease, she was unable to wear a bra and wore a sports bra all day, even sleeping in one.

[0485] Past medical history

[0486] Regular 28-day cycles, no pregnancy, severe endometriosis, multiple surgeries, and no abnormal blood parameters noticed.

[0487] VAS pain scale before onset: 100mm

[0488] treat

[0489] As described in Example 1, subcutaneous implantation of 80 mg testosterone and 4 mg anastrozole pellets was performed for 3 months, repeated for two additional 3-month periods, followed by breast augmentation, and then two more 3-month postoperative treatments as described above.

[0490] result

[0491] i. VAS pain scale at 4 weeks: 0 mm

[0492] ii. No inflammation was detected at 4 weeks.

[0493] iii. Fibroglandular tissue decreased by 36%.

[0494] iv. Breast volume decreased by 23%, returning to its pre-illness size.

[0495] v. Complete reversal of extreme background enhancement in MRI

[0496] vi. Without SAE

[0497] vii. BMI remained unchanged.

[0498] MRI images of the patient's breasts before and after treatment, as shown below. Figure 22 As shown in Figure A, and the mammograms of the patient's breasts before and after treatment are as follows: Figure 22 As shown in Figure B. Breast volume and density measurements are as follows: Figure 22 As shown in C.

[0499] Example 8: Treatment of Autoimmune Inflammatory Mastitis

[0500] patient: 42.04033771

[0501] Medical visit

[0502] A 24-year-old physical therapist experienced rapid breast enlargement—"extreme pain," diffuse inflammation, and persistent, severe pain for four months—making her unable to work and unresponsive to the following:

[0503] i. Over-the-counter analgesics and NSAIDs

[0504] ii.OCP

[0505] iii. Oral progesterone

[0506] iv. Danazol

[0507] v.LhRh agonist

[0508] Bra cup size

[0509] The tumor was 36B before the onset of the disease, and increased to 36EE.

[0510] Past medical history

[0511] The patient's obstetric / gynecological history includes irregular 28-day cycles—amenorrhea due to Zoladex, and no pregnancy. No abnormalities were noted in blood parameters.

[0512] VAS pain scale before onset: 100mm

[0513] treat

[0514] Subcutaneous implantation of 100 mg testosterone and 3 mg anastrozole pellets for 3 months followed by breast reduction surgery, and then two more of the above postoperative treatments for 3 months.

[0515] result

[0516] i. VAS pain scale 50 mm at 4 weeks

[0517] ii. Inflammation significantly decreased at 4 weeks.

[0518] iii. Reversal of high-density mammograms and extreme background-to-solid enhancement on MRI

[0519] iv. No SAE

[0520] v. BMI remained unchanged.

[0521] vi. Subsequently diagnosed with myasthenia gravis

[0522] Mammogram images of the patient's breasts before treatment, such as Figure 23 As shown in Figure A. Post-treatment MRI images are as follows. Figure 23 As shown in B.

[0523] Example 9: Treatment of plasma cell mastitis

[0524] Plasma cell mastitis is an autoimmune inflammatory process of destruction of the mammary ducts behind the areola, resulting in multiple fistulas and inevitably leading to disfiguring surgery and a high risk of recurrence. There is no known treatment for this condition other than surgery, which has significant limitations. An example of plasma cell mastitis is the invasion of the mammary ductal system by inflammatory cells that secrete pro-inflammatory cytokines. The IL-6 inflammatory pathway has recently been shown to be crucial in this inflammatory process and has been targeted as a potential mechanism for treating plasma cell mastitis (Liu, 2020). It has been demonstrated that the female breast contains cells that respond dramatically (53%) to testosterone treatment in reducing IL-6 levels (Guhl, 2012).

[0525] A 43-year-old otherwise healthy woman developed multiple fistulas in the right nipple region of the areolar complex. Initially unresponsive to antibiotic therapy, she received only short-term relief with high-dose corticosteroids and a surgical total ductectomy. Significant inflammation was present around the right nipple-areolar complex, with four fistulas at the 4 o'clock position on the areolar rim. Significant pain (8) was measured on a 0–10 cm visual analog scale. She began T+AI therapy and received subcutaneous implants of 80 mg testosterone and 4 mg anastrozole. Within three weeks, her visual analog scale score decreased to 3, and the periareolar redness also subsided. After 11 months of inserting three implants of the same concentration, the fistulas completely resolved without associated pain. A 12-month follow-up showed no evidence of disease recurrence.

[0526] Example 10: Granulomatous mastitis

[0527] Because the inflammation occurs around the lobules of the breast, granulomatous mastitis is also known as granulomatous lobular mastitis. It is a manifestation of autoimmune inflammatory mastitis in the breast tissue, leading to granuloma formation, inflammation, and fistula formation. There is no known cure for this condition, and women often undergo multiple surgeries and / or immunosuppressive therapy.

[0528] A 32-year-old woman sought a second opinion after six months of treatment for histologically confirmed granulomatous mastitis. She had been given antibiotics, corticosteroids, methotrexate, and underwent surgery to remove a fistula. All of this 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.

[0529] Between the first and second MRI scans taken on May 23, 2018, she had three implants inserted, as described in Example 1, containing 80 mg of testosterone and 4 mg of anastrozole. The inflammation rapidly subsided, and the remaining fistula healed over a three-month period. Tenderness and breast discomfort persisted for five months before slowly receding. A clinical examination on May 23, 2018, showed only a minor scar on her breast from the previous surgery, with no other abnormalities. A follow-up examination in February 2020 showed she was still asymptomatic.

[0530] Mammogram images of the patient's breasts before treatment, such as Figure 24 As shown in Figure A. The MRI images obtained after treatment are as follows. Figure 23 As shown in B.

[0531] Other exemplary and non-limiting embodiments.

[0532] Further advantages of the claimed subject matter will become apparent from the following examples describing certain embodiments of the claimed subject matter.

[0533] Example A

[0534] 1A. A pharmaceutical preparation comprising:

[0535] Effective amounts of androgens, effective amounts of aromatase inhibitors, and binding agents;

[0536] The formulation, when administered to a subject, provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the androgen and plasma concentrations of the aromatase inhibitor; and

[0537] The sustained release multiphase concentration pattern in the subject's serum or plasma includes:

[0538] In the first time period, the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor has an increased concentration in plasma but below its Tmax concentration in plasma; and

[0539] In the second time period, the androgen initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

[0540] 2A. A pharmaceutical preparation comprising:

[0541] Effective amounts of androgens, effective amounts of aromatase inhibitors, and binding agents;

[0542] The pharmaceutical preparation is compressed into small pellets;

[0543] When the pellet is administered subcutaneously to a subject, it provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the androgen and plasma concentrations of the aromatase inhibitor; and

[0544] The sustained release multiphase concentration pattern in the subject's serum or plasma includes:

[0545] In the first time period, the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor has an increased concentration in plasma but below its Tmax concentration in plasma; and

[0546] In the second time period, the androgen initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

[0547] 3A. A pharmaceutical formulation comprising:

[0548] Effective amounts of androgens, effective amounts of aromatase inhibitors, and binding agents;

[0549] The formulation, when administered to a subject, provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the androgen and plasma concentrations of the aromatase inhibitor; and

[0550] The sustained release multiphase concentration pattern in the subject's serum or plasma includes:

[0551] In the first time period, the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor has an increased concentration in plasma but is below its Tmax concentration in plasma;

[0552] In the second time period, the androgen initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

[0553] In the third time period, the androgen has a second peak concentration in serum below Tmax, and the aromatase inhibitor gradually decreases in plasma concentration, falling below the androgen concentration during the third time period; and

[0554] In the fourth time period, the serum concentration of the androgen gradually decreased, and the plasma concentration of the aromatase inhibitor gradually decreased, with the decrease in both being approximately the same.

[0555] 4A. A pharmaceutical preparation comprising:

[0556] Effective amounts of androgens, effective amounts of aromatase inhibitors, and binding agents;

[0557] The pharmaceutical preparation is compressed into small pellets;

[0558] When the pellet is administered subcutaneously to a subject, it provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the androgen and plasma concentrations of the aromatase inhibitor; and

[0559] The sustained release multiphase concentration pattern in the subject's serum or plasma includes:

[0560] In the first time period, the androgen has a first peak concentration (Tmax) in serum and the aromatase inhibitor has an increased concentration in plasma but is below its Tmax concentration in plasma;

[0561] In the second time period, the androgen initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

[0562] In the third time period, the androgen has a second peak concentration in serum below Tmax, and the aromatase inhibitor gradually decreases in plasma concentration, falling below the androgen concentration during the third time period; and

[0563] In the fourth time period, the serum concentration of the androgen gradually decreased, and the plasma concentration of the aromatase inhibitor gradually decreased, with the decrease in both being approximately the same.

[0564] 5A. A pharmaceutical preparation comprising:

[0565] 60 mg to 120 mg of testosterone or its esters, 4 mg to 6 mg of aromatase inhibitor and stearic acid;

[0566] The pharmaceutical preparation is compressed into pellets with a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm;

[0567] When the pellet is administered subcutaneously to a subject, it provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the testosterone or its esters and plasma concentrations of the aromatase inhibitor; and

[0568] The sustained release multiphase concentration mode includes:

[0569] In the first time period, the testosterone or its ester has a first peak concentration (Tmax) in serum and the aromatase inhibitor has an increased concentration in plasma but below its Tmax concentration in plasma; and

[0570] In the second time period, the testosterone or its ester initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

[0571] 6A. A pharmaceutical preparation comprising:

[0572] 60 mg to 120 mg of testosterone or its esters, 4 mg to 6 mg of aromatase inhibitor and stearic acid;

[0573] The pharmaceutical preparation is compressed into pellets with a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm;

[0574] When the pellet is administered subcutaneously to a subject, it provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the testosterone or its esters and plasma concentrations of the aromatase inhibitor; and

[0575] The sustained release multiphase concentration mode includes:

[0576] In the first time period, the testosterone or its ester 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;

[0577] In the second time period, the testosterone or its ester initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

[0578] In the third time period, the testosterone or its ester has a second peak concentration in serum below Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases, and during the third time period, drops to a concentration below that of the testosterone or its ester; and

[0579] In the fourth time period, the serum concentration level of the testosterone or its ester gradually decreased, and the plasma concentration of the aromatase inhibitor gradually decreased, with the decrease levels of both being approximately the same.

[0580] 7A. Pharmaceutical formulations of Examples 1A, 2A, or 5A, wherein the sustained-release multiphase concentration pattern further includes:

[0581] In the third time period, the testosterone or its ester has a second peak concentration in serum below Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases, and during the third time period, drops to a concentration below that of the testosterone or its ester; and

[0582] In the fourth time period, the serum concentration level of the testosterone or its ester gradually decreased, and the plasma concentration of the aromatase inhibitor gradually decreased, with the decrease levels of both being approximately the same.

[0583] 8A. A pharmaceutical formulation of one or more of Examples 1A to 7A, wherein the aromatase inhibitor exhibits primary release during the first time period.

[0584] 9A. Pharmaceutical formulations of one or more of Examples 1A to 8A,

[0585] During the first time period, the aromatase inhibitor did not exhibit zero-order release.

[0586] 10A. A pharmaceutical formulation of one or more of Examples 1A to 9A, wherein the aromatase inhibitor does not exhibit zero-order release during the second time period.

[0587] 11A. A pharmaceutical formulation of one or more of Examples 1A to 10A, wherein the testosterone or its esters do not exhibit zero-order release during the second time period.

[0588] 12A. A pharmaceutical formulation of one or more of Examples 1A to 11A, wherein the aromatase inhibitor does not exhibit zero-order release during the third time period.

[0589] 13A. A pharmaceutical formulation of one or more of Examples 1A to 12A, wherein the testosterone or its esters do not exhibit zero-order release during the third time period.

[0590] 14A. A pharmaceutical formulation of one or more of Examples 1A to 13A, wherein the aromatase inhibitor exhibits primary release during the third time period.

[0591] 15A. A pharmaceutical formulation of one or more of Examples 1A to 14A, wherein during the third time period, the testosterone or its ester exhibits primary release.

[0592] 16A. A pharmaceutical formulation of one or more of Examples 1A to 15A, wherein the aromatase inhibitor does not exhibit zero-order release during the fourth time period.

[0593] 17A. A pharmaceutical formulation of one or more of Examples 1A to 16A, wherein the testosterone or its esters do not exhibit zero-order release during the fourth time period.

[0594] 18A. A pharmaceutical formulation of one or more of Examples 1A to 17A, wherein the aromatase inhibitor exhibits primary release during the fourth time period.

[0595] 19A. A pharmaceutical formulation of one or more of Examples 1A to 18A, wherein during the fourth time period, the testosterone or its ester exhibits primary release.

[0596] 20A. A pharmaceutical formulation of one or more of Examples 1A to 19A, wherein the first time period ends immediately after the androgen, having a first peak concentration (Tmax) in serum.

[0597] 21A. A pharmaceutical preparation of one or more of Examples 1A to 20A, wherein the first time period ends in 5 to 14 hours.

[0598] 22A. A pharmaceutical preparation of one or more of Examples 1A to 21A, wherein the first time period ends between 5.5 hours and 13 hours.

[0599] 23A. A 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.

[0600] 24A. A pharmaceutical preparation of one or more of Examples 1A to 23A, wherein the second time period ends between 23 hours and 80 hours.

[0601] 25A. A pharmaceutical preparation of one or more of Examples 1A to 24A, wherein the pharmaceutical preparation is an implant.

[0602] 26A. A pharmaceutical preparation of one or more of Examples 1A to 25A, wherein the implant is a compressed pellet.

[0603] 27A. A pharmaceutical preparation of one or more of Examples 1A to 24A, wherein said pharmaceutical preparation is a transdermal patch.

[0604] 28A. A pharmaceutical formulation of one or more of Examples 1A to 25A, wherein the implant is administered subcutaneously to a subject.

[0605] 28A. A pharmaceutical preparation of one or more of Examples 1A to 26A, wherein the compressed pellet is administered subcutaneously to a subject.

[0606] Example B

[0607] 1B. A pharmaceutical preparation comprising:

[0608] Approximately 80 mg of testosterone or its esters, approximately 4 mg of anastrozole, and approximately 2 mg of stearic acid;

[0609] The pharmaceutical preparation is compressed into pellets with a diameter of 4.4 mm to 4.6 mm and a length of 4 mm to 7 mm;

[0610] When the pellet is administered subcutaneously to a subject, it provides a sustained, multiphase release pattern of concentrations in the subject's blood over time, as measured by serum concentrations of the testosterone or its esters and plasma concentrations of the anastrozole; and

[0611] The sustained release multiphase concentration mode includes:

[0612] In the first time period, the testosterone or its ester has a first peak concentration (Tmax) in serum and the concentration of anastrozole in plasma increases but is below its Tmax concentration in plasma;

[0613] In the second time period, the testosterone or its ester initially has a decreased serum concentration level and then an increased serum concentration level, and the anastrozole has its Tmax concentration in plasma.

[0614] In the third time period, the testosterone or its ester has a second peak concentration in serum below Tmax, and the concentration of anastrozole in plasma gradually decreases, and during the third time period, drops to a concentration below that of the testosterone or its ester; and

[0615] In the fourth time period, the serum concentration level of the testosterone or its ester gradually decreased, and the plasma concentration of the anastrozole gradually decreased, with the decrease levels of both being approximately the same.

[0616] While 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 thereby cover the methods and structures within the scope of these claims and their equivalents.

[0617] In the above description of some embodiments, specific terms have been used for clarity. However, this disclosure is not intended to be limited to the specific terms so chosen, and it should be understood that each specific term includes other technical equivalents that operate in a similar manner to achieve similar technical purposes. Terms such as “left” and “right,” “front” and “rear,” “up” and “down” are used for convenience to provide reference points and should not be construed as limiting terms.

[0618] In this specification, the word “comprise” should be understood in its “open” sense, that is, in the sense of “including”, and therefore is not limited to its “closed” sense, that is, “consisting of only”. When it appears, the corresponding meaning should be attributed to the corresponding words “comprise”, “comprised”, and “comprises”.

[0619] Furthermore, only some embodiments of the present invention have been described above, and changes, modifications, additions and / or variations may be made to them without departing from the scope and spirit of the disclosed embodiments. These embodiments are exemplary and not restrictive.

[0620] It should be understood that the present invention is not limited to the disclosed embodiments, but rather is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the invention. Furthermore, 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. Moreover, each individual feature or component of any given component can constitute an additional embodiment.

Claims

1. Use of an androgen agent and an aromatase inhibitor in the preparation of a medicament for the prevention or treatment of autoimmune inflammatory mastitis in patients with such need, said medicament comprising i) an effective amount of an androgen agent and ii) an effective amount of an aromatase inhibitor, said aromatase inhibitor being used to block the conversion of the androgen agent into estrogen by aromatase, said medicament being a solid, sustained-release pharmaceutical formulation having a sustained-release multiphasic concentration pattern over time in the patient's blood, as measured by serum concentrations of said androgen agent and plasma concentrations of said aromatase inhibitor, wherein, The drug is intended for subcutaneous delivery to the patient, wherein the aromatase inhibitor is anastrozole, the androgen is testosterone or a pharmaceutically acceptable salt or ester thereof, the ester including heptaester, propionate, cyclopentylpropionate, phenylacetate, isobutyrate, cyclobutyl ester, and undecanoate, and the sustained release multiphasic concentration pattern in the patient's serum or plasma includes: In the first time period, the androgen agent has a first peak concentration (Tmax) in serum and the aromatase inhibitor has an increased concentration in plasma but below its Tmax concentration in plasma; and In the second time period, the androgen initially has a decreased serum concentration level followed by an increased serum concentration level, and the aromatase inhibitor has its Tmax concentration in plasma.

2. The use according to claim 1, wherein the drug is in pellet form with a hardness between 6 kg / N and 10 kg / N to provide a multiphase release mode of the pellet.

3. The use according to claim 1 or 2, wherein the drug is a pharmaceutical preparation comprising: Effective amounts of androgens, effective amounts of aromatase inhibitors, and binding agents; and The pharmaceutical formulation is compressed into pellets with a hardness of approximately 8 kg / N to provide a multiphase release mode of the pellets.

4. The use according to claim 1 or 2, The sustained release multiphase concentration pattern in the patient's serum or plasma further includes: In the third time period, the androgen agent has a second peak concentration in serum below Tmax, and the concentration of the aromatase inhibitor in plasma gradually decreases, and during the third time period, drops to a concentration below that of the androgen agent; and In the fourth time period, the serum concentration level of the androgen gradually decreased, and the plasma concentration of the aromatase inhibitor gradually decreased, with the decrease levels of both being approximately the same.

5. The use according to claim 4, wherein the aromatase inhibitor does not exhibit zero-order release during the third time period.

6. The use according to claim 4, wherein the aromatase inhibitor exhibits primary release during the third time period.

7. The use according to claim 4, wherein during the third time period, the androgen agent does not exhibit zero-order release.

8. The use according to claim 4, wherein during the third time period, the androgen agent exhibits primary release.

9. The use according to claim 4, wherein the aromatase inhibitor does not exhibit zero-order release during the fourth time period.

10. The use according to claim 4, wherein during the fourth time period, the androgen agent does not exhibit zero-order release.

11. The use according to claim 4, wherein during the fourth time period, the aromatase inhibitor exhibits primary release.

12. The use according to claim 4, wherein during the fourth time period, the androgen agent exhibits primary release.

13. The use according to claim 1 or 2, wherein the aromatase inhibitor exhibits primary release during the first time period.

14. The use according to claim 1 or 2, wherein the aromatase inhibitor does not exhibit zero-order release during the first time period.

15. The use according to claim 1 or 2, wherein the aromatase inhibitor does not exhibit zero-order release during the second time period.

16. The use according to claim 1 or 2, wherein the androgen agent does not exhibit zero-order release during the second time period.

17. The use according to claim 1 or 2, wherein the first time period ends immediately after the androgen agent has a first peak concentration (Tmax) in serum.

18. The use according to claim 1 or 2, wherein the first time period ends between 5 hours and 14 hours.

19. The use according to claim 1 or 2, wherein the first time period ends between 5.5 hours and 13 hours.

20. The use according to claim 1 or 2, wherein the second time period ends immediately after the aromatase inhibitor has its Tmax.

21. The use according to claim 1 or 2, wherein the second time period ends between 23 hours and 80 hours.

22. The use according to claim 1 or 2, wherein, The drug is a pharmaceutical preparation, which includes: 60 mg to 120 mg of testosterone or its esters, 2 mg to 6 mg of aromatase inhibitor, and stearic acid; and The pharmaceutical preparation is compressed into pellets with a diameter of 4.25 mm to 4.75 mm and a length of 4 mm to 7 mm.

23. The use according to claim 1 or 2, wherein the autoimmune inflammatory mastitis is selected from the group consisting of idiopathic inflammatory macromastia, plasma cell mastitis, granulomatous mastitis, and combinations thereof.

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