WO2009036566A1 - Methods for detecting abnormalities in breast tissue using aromatase inhibitors - Google Patents

Abstract

The invention relates to improved methods for detecting abnormalities in breast tissue, in particular dense breast tissue. A method of the invention generally comprises administering to a subj ect an effective amount of one or more aromatase inhibitor to reduce breast density and/or vascularity in the subject prior to making an image of the breast tissue.

Description

Title: Methods for Detecting Abnormalities in Breast Tissue Using Aromatase

Inhibitors

FIELD OF THE INVENTION

The invention generally relates to methods and formulations for detecting abnormalities in breast tissue.

BACKGROUND OF THE INVENTION

Breast cancer is the most common form of cancer in women, and is the second leading cause of cancer death in humans. It is well known that early detection of abnormalities combined with appropriate medical care reduces the risk of death or severe medical consequences. Mammography, along with physical breast examination, is the modality of choice for screening for early breast cancer. Magnetic resonance imaging (MRI) is a highly effective emerging imaging technique in the diagnosis and staging of breast cancer (Bedrosian, 2003). MRI has been reported to be particularly accurate in detecting high grade tumors (Bedrosian, 2003) and it is significantly more sensitive than mammograms or breast ultrasound in diagnosing breast cancer in BRCA mutation carriers (Hagen, 2007). However, the accuracy of a breast imaging method depends on a number of factors including breast density and vascularity which may result in false-positive or false- negative results. SUMMARY OF THE INVENTION The invention relates to improved methods for detecting abnormalities in breast tissue, in particular dense breast tissue. A method of the invention generally comprises administering to a subject an effective amount of one or more aromatase inhibitor (AI) to reduce breast density and/or vascularity in the subject prior to making an image of the breast tissue. In an aspect, the invention relates to a method for reducing breast density and/or vascularity in a subject comprising administering to the subject an effective amount of one or more AI to reduce breast density and/or vascularity prior to making an image of breast tissue of the subject.

In another aspect, the invention provides a method for detecting an abnormality in breast tissue in a subject comprising administering to the subject an effective amount of one or more AI to reduce the density and/or the vascular perfusion of the breast tissue in the subject, and making an image of the breast tissue such that the abnormality in the breast is capable of being, or is detected.

An abnormality in breast tissue includes without limitation, a mass, a cyst, a microcalcification, a fibrous finding, an architectural distortion, and/or other abnormal findings related to benign or malignant abnormalities. In particular aspects of the invention, the abnormality in the breast tissue is a breast lesion, more particularly a tumor, which may be benign or malignant. In particular aspects of the invention the abnormality in the breast tissue is a high grade tumor.

An image of breast tissue may be made using techniques known in the art, including, without limitation, film mammography, digital mammography, magnetic resonance imaging (MRI), dynamic contrast-enhanced MRI, or alternative techniques for visualizing breast tissue such as ultrasonography, thermography, computed tomography, positron emission tomography, transillumination, or radionucleotide imaging.

The invention also relates to a packaged formulation for reducing breast density and/or vascularity in a subject prior to making an image of the subject's breast tissue comprising one or more AI packaged with instructions for using the AI(s) for reducing breast density and/or vascularity in the subject prior to making an image of the subject's breast tissue.

The invention involves the use of one or more AI, either alone or in combination with other pharmaceutical agents (e.g. hormone replacement therapy) for reducing breast density and/or vascularity in a subject before making an image of the subject's breast tissue.

The invention also involves the use of one or more AI for preparation of a packaged formulation for reducing breast density and/or vascularity in a subject prior to making an image of the subject's breast tissue. In an aspect, the invention relates to the use of an aromatase inhibitor for use in combination with mammography or MRI or for use in the manufacture of a medicament for use in combination with mammography or MRI in a subject.

Other objects, features and advantages of the present invention will become apparent from the following drawings and detailed description. BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows scanned images acquired through three different sources, a. Digital image acquired by Direct Digital Acquisition (DDA); b. Image scanned with Kodak L S85 film digitizer; and c. Image scanned with Agfa DuoScan Flatbed scanner.

Figure 2 shows the editing tools included in the ImageQuant 5 program. Figure 3 shows selection and background correction in respect to a comparison of scanned images using the ImageQuant 5 program.

Figure 4 shows the outlining of the total area of glandular tissue and the densest glandular region of a scanned image using the ImageQuant 5 program.

Figure 5 shows a visual representation of breast density using the "Surface Plot" function of the Image J program.

Figure 6 shows a 3d interactive surface plot of breast density that is available as a plugin of the Image J program.

Figure 7 shows a display of a greyscale image in colors through the "lookup tables" tool of the Image J program. Greyscale images are displayed using a color lookup table which describes the color to be used for each of 256 possible displayed pixel values.

Figure 8 shows a comparison of breast density for a patient receiving an aromatase inhibitor pre- or post- mammogram.

Figure 9 shows a comparison of breast density for a patient receiving an aromatase inhibitor pre- or post- mammogram. Figure 10 shows a 3-D interactive surface plot of pixel density of images taken from a patient before and after receiving an aromatase inhibitor.

Figure 11 shows a comparison of breast density in a subject receiving an aromatase inhibitor pre- or post- mammogram.

Figure 12 shows MRI-Tl : A pre-contrast, B post-contrast and C subtraction views.

Figure 13 shows benign background enhancement in MRI and its Image J interactive surface plotting, a) MRI benign breast tissue with no background enhancement after gadolinium injection, b) Interactive surface plotting of Image J representation of the breast in picture a), c) MRI stippled foci of benign background enhancement in the same patient, d) Interactive surface plotting of Image J representation of the breast in picture c) assigning lighter gradient of color corresponding to the increased pixel intensity caused by the stippled foci of enhancement. - A -

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

The invention is related to methods, formulations and compositions for improved detection of breast abnormalities, in particular breast abnormalities in dense breast tissue.

The methods of the invention generally comprise administering to a subject an effective amount of one or more AI to reduce breast density and/or vascularity in the subject prior to making an image of the subject's breast tissue.

For convenience, certain terms employed in the specification and claims are collected below.

The term "subject" refers to a warm-blooded animal such as a mammal, which requires a reduction in breast density and/or vascularity, and/or is being subjected to methods for imaging breast tissue, for example, mammography or MRI. In particular, the term refers to a human, more particularly a female human. In embodiments of the invention the subject is a menopausal female, in particular a menopausal female receiving hormone replacement therapy. In embodiments of the invention the subject is a postmenopausal female. In aspects of the invention the term includes male humans. In aspects of the invention, a subject is a BRCA mutation carrier. In other aspects, the subject has multiple cancerous foci in breast tissue. In other aspects, the subject has a high incidence of interval breast malignancies (Komenaka, 2004).

An "aromatase inhibitor" refers to a substance that inhibits or inactivates aromatase, an enzyme of the cytochrome P-450 superfamily and a product of the CYPl 9 gene. The enzyme synthesizes estrogens from androgenic substrates, for example, estrone is synthesized from the preferred substrate androstenedione and estradiol from testosterone. The inhibitors have been classified as first-, second- and third- generation inhibitors according to the chronologic order of their clinical development, and as type 1 or type 2 inhibitors according to their mechanism of action. (See, for example, Smith, M.D., I.E. and Dowsett, Ph.D., M, N Eng J Med 2003, 348, 2431-2442 for a discussion of aromatase inhibitors). Generally, Type 1 aromatase inhibitors are steroidal analogues of androstenedione that bind irreversibly to aromatase to thereby inactivate the enzyme, and Type 2 aromatase inhibitors are nonsteroidal and bind reversibly to the heme group of the enzyme by way of a basic nitrogen.

In aspects of the invention, the aromatase inhibitors for use in the methods of the present invention include the compounds described in U.S. Patent Nos. 4,808,616, 4,904,650 and 4,322,416; EP Patent Publication Nos. EP-A-165904, EP-A-236940, EP-A- 408509, EP 91810110.6, EP-A-114033, EP-A-166692, EP-A-356673, EP-A-337929, EP- A-337928, EP-A-340153, EP-A-293978, EP-A-250198, EP-A-281283, EP-A-296749, EP- A-299683, EP-A-299684, EP-A-316097, EP-A-354689, EP-A-354683, andEP-A-181287; Swiss Patent Application Nos. 1339/90-7, 3014/90-0, 3013/90-0, 3923/90-4; German Patent Application Nos. DE-A-4014006, DE-A-3926365, DE-A-3740125, and DE-A- 3622841 ; or Published British Patent Application No. GB-A-2171100.

Examples of aromatase inhibitors include: 6-methylenandrosta-l,4-diene-3,17- dione; 1 -methyl-6-methylenandrosta- 1 ,4-diene-3 , 17-dione; 1 -ethyl-6-methylenandrosta- l,4-diene-3,l 7-dione; 4-methyl-6-methylenandrosta-l,4-diene-3,l 7-dione; 4-ethyl-6- methylenandrosta- 1 ,4-diene-3 , 17-dione; 4-fluoro-6-methylenandrosta- 1 ,4-diene-3 , 17- dione; 4-chloro-6-methylenandrosta- 1 ,4-diene-3 , 17-dione; 6-ethylidenandrosta- 1 ,4-diene- 3 , 17-dione; 6-propylidenandrosta- 1 ,4-diene-3 , 17-dione; 4-fluoro- 1 -methyl-6- methylenandrosta- 1 ,4-diene-3 , 17-dione; 4-chloro- 1 - methyl-6-methylenandrosta- 1 ,4-diene- 3,17-dione; l-methyl-6-ethylidenandrosta-l,4-diene-3,l 7-dione; and 4-fluoro-6- ethylidenandrosta-l,4-diene-3, 17- dione.

In aspects of the invention, an aromatase inhibitor is 6-methylenandrosta-l ,4-diene- 3,17-dione, also known as exemestane (Aromasin®). 6-Methylenandrosta-l,4-diene-3,17- dione may be synthesized using the process of Example 1 in U.S. Patent Nos. 4,808,616 and 4,904,650. Other examples of aromatase inhibitors useful in the methods and formulations of the invention include: 3-(4-aminophenyl)-3-ethyl-2, 6- piperidinedione also known as aminoglutethimide (Cytadren®); 4-hydroxyandrostenedione, also known as formestane; 1 -methylandrosta-1 ,4-diene-3, 17-dione, also known as atamestane; α,α,α, α'- tetramethyl-5-(l H-l,2,4-triazol-l-ylmethyl)-l,3-benzenediacetonitrile, also known as anastrazole (see U.S. RE36617) and marketed as Arimidex®; 4-(5,6,7,8- tetrahydroimidazo[l,5-a]pyridin-5-yl)-benzonitrile monohydrochloride, also known as fadrozole; 4-(3-(4-Fluorophenyl)-2-hydroxy- 1 -( 1 H- 1 ,2,4-triazol- 1 -yl)-propyl)- benzonitrile also known as finrozole; 4,4'-(lH-l,2,4-triazol-l-ylmethylene)bis-benzonitrile, also known as letrozole (see U. S. Patent 4,978,672) and marketed as Femara®; 6-[(4- chlorophenyl)- 1 H- 1 , 2,4- triazol- 1 -ylmethyl]- 1 -methyl- 1 H- benzotriazole, also known as vorozole; and 4- [N-(4-bromobenzyl)-N-(4-cyanophenyl)amino]-4H-l ,2,4- triazole. An aromatase inhibitor may be formulated with a pharmaceutically acceptable carrier, excipient or vehicle. The term "pharmaceutically acceptable carrier, excipient, or vehicle" refers to a medium which does not interfere with the effectiveness or activity of an active ingredient and which is not toxic to the hosts to which it is administered. A carrier, excipient, or vehicle includes diluents, binders, adhesives, lubricants, disintegrates, bulking agents, wetting or emulsifying agents, pH buffering agents, and miscellaneous materials such as absorbants that may be needed in order to prepare a particular composition. The use of such media and agents for an active substance is well known in the art. Suitable pharmaceutical carriers, excipients, and vehicles are described in the standard text, Remington: The Science and Practice of Pharmacy, [(formerly called Remington's Pharmaceutical Sciences), Editor: University of the Sciences in Philadelphia (USIP), 21^st Edition, May, 2005].

It is contemplated that aromatase inhibitors may be administered to a subject as a pharmaceutically acceptable salt, prodrug, or a salt of a prodrug. The aromatase inhibitors may also be in the form of hydrates or their crystals and may include the solvent used for crystallization. Further the aromatase inhibitors may occur in the form of R- or S- enantiomers and as enantiomeric mixtures thereof, for example in the form of a racemate. Isomers of aromatase inhibitors, mixtures of at least two isomers (e.g. mixtures of diastereoisomers or enantiomers), and geometric isomers (e.g., cis- and trans-isomers) may also be utilized in the present invention. All forms and variations of aromatase inhibitors are intended to be included in the invention.

It is also contemplated that the aromatase inhibitor is a sustained release formulation, i.e, a formulation that allows for the continuous delivery of an AI to a subject over a period of time (e.g., 1-2 days, 1-3 days, 1-5 days, 1-7 days, 1-2 weeks, 1-3 weeks, 1- 4 weeks, 1-5 weeks, 1-7 weeks, 1-10 weeks, 1-12 weeks, 1-9 weeks, 1-6 weeks, 1 -12 months, 1-9 months, 1-6 months or 1-3 months).

The term "breast density" refers to the appearance of the breast parenchyma produced using techniques for imaging breast tissue. Breast density can be classified using the system proposed by Wolfe, JN, Cancer, 37:2486-2492, 1976 which is based on the appearance of four breast parenchyma patterns in mammograms (i.e., the categories Nl, Pl, P2 and DY). InNl, the breast consists of mostly fatty tissue with no ducts visible and it represents an essentially normal breast. The Pl category represents a fatty breast, with predominant ducts in the anterior portion occupying up to a quarter of the breast area. In P2, the breast is involuted, with prominent duct patterns of moderate to severe degree, occupying more than a quarter of the breast, and the visible duct pattern may occupy the entire breast. In DY, the breast parenchyma is dense, which usually denotes connective tissue hyperplasia, and it may appear homogeneous due to the overall increased density. Breast tissues classified as P2 or DY using the Wolfe system are generally considered to be "dense breasts".

The American College of Radiology (ACR) has proposed a modified version of the Wolfe patterns for the BI-RADS [American College American College of Radiology. Illustrated Breast Imaging Reporting and Data System (BI-RADS), The American College of Radiology, third edition, 1998]. The classification scheme is generally as follows:

I. The breast is almost entirely fat.

II. There are scattered fibroglandular densities.

III. The breast is heterogeneously dense. This may lower the sensitivity of mammography.

IV. The breast tissue is extremely dense, which could obscure a lesion in mammography.

Other pattern classifications include, without limitation the six-category classification (SCC) and others (see, for example, Marias K, et al, Conf Proc IEEE Eng Med Biol Soc. 6:6394-8, 2005; and Jeffreys, M., Br J Radiol. 76(908):561-3, 2003 and Table 3 herein).

The term "vascularity" includes vessel permeability, vascular volume and perfusion of breast tissue.

The phrases "to reduce breast density and/or vascularity" or "for reducing breast density and/or vascularity" refer to methods and compositions that result in a change in breast density and/or vascularity. The terms include a change in the breast parenchyma classification of a subject to a less dense or more lucent appearance and/or a change in the vascularity of breast tissue of a subject, relative to the breast density and/or vascularity observed in the subject in the absence of a treatment with a method or formulation of the invention. In MRI, the terms may include a reduction in baseline enhancement of contrast material (e.g. Gadolinium dye) uptake by the blood vessels. The terms "making an image of breast tissue", "imaging breast tissue" and "making an image of the subject's breast tissue" refer to one of a variety of techniques known in the art for visualizing breast tissue. Suitable techniques include, without limitation, film mammography, digital mammography, ultrasonography, thermography, computed tomography, positron emission tomography, transillumination, magnetic resonance imaging, contrast-enhanced MRI, or radionucleotide imaging (see for example, Jackson, VP et al, Radiology 188:297-301, 1993; Braeuning, MP et al, Breast Cancer Res. Treat. 35:31-38, 1995; Lehman CD, Journal of Magnetic Resonance Imaging 24 (5): "964-70, 2006; Helvie MA. Imaging analysis: Mammography. In: Harris JR, LippmanME, Morrow M, Osborne CK, eds. Diseases of the Breast. 3rd ed. Philadelphia, Pa: Lippincott Williams & Wilkins; 2004:103-130; Amercian Cancer Society, Mammograms and Other Breast Imaging Procedures http://www.cancer.org/docroot/PED/content/PED 2 3X Mammography and Other Breast Imaging Procedures.asp; Stomper PC, et al, Breast Cancer Res Treat 1997;45 : 39-46; Fischer DR, et al, AJR 2006; 187:147 -151). In aspects of the invention, an image is made using film mammography. In other aspects of the invention an image is made using digital mammography. In other aspects of the invention an image is made using MRI.

The term "abnormality of the breast" refers to any abnormality in breast tissue that may be detected by making an image of the breast tissue. An abnormality in breast tissue includes without limitation, a mass, a cyst, a microcalcification, a fibrous finding, an architectural distortion, and/or other abnormal findings related with benign or malignant abnormalities. In particular aspects of the invention, the abnormality in the breast tissue is a breast lesion, more particularly a tumor, which may be benign or malignant. In particular aspects of the invention the abnormality is a high grade tumor. The invention relates to a method for reducing breast density and/or vascularity in a subject prior to making an image of breast tissue of the subject. The method involves administering an effective amount of one or more AI to reduce breast density in the subject.

The invention also relates to methods for detecting an abnormality of the breast of a subject. The methods comprise administering to the subject an effective amount of one or more AI to reduce breast density and/or vascularity of the subject's breast tissue, and making an image of the breast tissue such that an abnormality of the breast can be or is detected. In embodiments of the invention, the abnormality is a malignant or benign tumor. In embodiments of the invention, the image of the breast tissue is made by standard film mammography. In other embodiments of the invention, the image of the breast tissue is made by digital mammography. In still other embodiments of the invention, the image of the breast tissue is made by MRI.

In aspects, the invention relates to a method for diagnosing breast cancer in a subject comprising administering an effective amount of an aromatase inhibitor to the subject and making an image of breast tissue of the subject to detect an abnormality of the breast tissue. In an aspect of the invention, a method for diagnosing breast cancer in a subject is provided comprising administering an effective amount of an aromatase inhibitor to the subject and subjecting the subject to mammography.

In another aspect of the invention, a method for diagnosing breast cancer in a subject is provided comprising administering an effective amount of an aromatase inhibitor to the subject and subjecting the subject to MRI. In other aspects, the invention relates to a method of preparing a female subject for mammography comprising the step of, prior to performing the mammography, administering to the subject one or more doses of at least one aromatase inhibitor, either alone or in conjunction with another pharmaceutical agent, in an effective amount to reduce breast density in the subject. In still other aspects, the invention relates to a method of preparing a female subject for breast MRI comprising the step of, prior to performing the breast MRI, administering to the subject one or more doses of at least one aromatase inhibitor, either alone or in conjunction with another pharmaceutical agent, in an effective amount to reduce breast vascularity in the subject. In particular aspects of this method, the subject is a BRCA mutation carrier. In other particular aspects of this method, the subject has multiple cancerous foci in breast tissue. In other particular aspects of this method, the subject has a high grade tumor. In other particular aspects of this method, the subject has a high incidence of interval breast malignancies (Komenaka, 2004).

An "effective amount" relates to the amount or dose of AI(s) that is sufficient to result in a desired reduction in breast density and/or to improve detection of an abnormality of the breast. The effective amount may vary with the particular subject involved, the density and/or vascularity of the breast tissue prior to treatment, the particular AI(s), and/or the intended result. In aspects of the invention, the amount of AI to reduce breast density and/or vascularity in a subject is in a dosage range of about 0.01 mg/kg to about 2000 mg/kg, 0.01 mg/kg to about 1000 mg/kg, 0.01 to about 500 mg/kg, 0.01 to about 250 mg/kg, 0.01 to about 200 mg/kg, 0.01 to about 100 mg/kg, 0.01 to about 75 mg/kg, 0.01 to about 60 mg/kg, 0.01 to about 50 mg/kg, 0.01 to about 25 mg/kg, or 0.01 to about 20 mg/kg, once, twice or three times daily. In other aspects of the invention, the amount of AI to reduce breast density and/or vascularity in a subject is in a dose of about 1 to about 500 mg, 10 to about 500 mg, 1 to about 100 mg, 1 to about 75 mg, 1 to 60 mg, 1 to about 50 mg, 2 to about 75 mg, 2 to about 60 mg, 2 to about 50 mg, 5 to about 75 mg, 5 to about 60 mg, 5 to about 50 mg, or 2 to about 20 mg once, twice or three times daily, preferably once daily.

In aspects of the invention, the dose of AI is about 0.01 mg/kg/day to about 400 mg/kg/day, 0.01 mg/kg/day to about 250 mg/kg/day, 0.01 mg/kg/day to about 200 mg/kg/day, 0.01 mg/kg/day to about 100 mg/kg/day, 0.01 mg/kg/day to about 75 mg/kg/day, 0.01 mg/kg/day to about 60 mg/kg/day, 0.01 mg/kg/day to about 50 mg/kg/day, 0.01 mg/kg/day to about 30 mg/kg/day, 0.01 mg/kg/day to about 20 mg/kg/day or 0.01 mg/kg/day to about 10 mg/kg/day.

By way of example, aminoglutethimide (marketed as Cytadren® and available as 250 mg oral tablets) may be administered initially at a dose of 250 mg given at 8-hour intervals and the dosage may be increased to a daily dose of 2 grams. Other commercially available aromatase inhibitors include letrozole, marketed as Femara®, which is available as a 2.5 mg oral tablet; anastrazole, marketed as Arimidex®, which is available as a 1 mg oral tablet and exemestane, marketed as Aromasin®, which is available as a 25 mg oral tablet. In aspects of the invention the aromatase inhibitor is letrozole which is administered in a daily dose of from about 2.5 mg to about 50 mg or 5 mg to about 20 mg, in particular 5 mg, 12.5 mg, or 20 mg. In an embodiment letrozole is administered for 1 to 3 days, preferably 3 days.

In other aspects, the aromatase inhibitor is anastrozole which is administered in a daily dose of from about 1 mg to about 100 mg, 1 mg to about 75 mg, or 1 mg to about 60 mg. In an embodiment anastrozole is administered for 1 to 3 days, preferably 3 days. In other aspects, the aromatase inhibitor is vorozole which is administered in a daily dose of from about 4 mg to about 100 mg, 4 mg to about 75 mg, 1 mg to about 60 mg or 4 mg to about 60 mg.

In further aspects, the aromatase inhibitor is exemestane which is administered in a daily dose of from about 25 mg to about 500 mg, 25 mg to about 250 mg, 10 mg to about 100 mg, 10 mg to about 50 mg, 25 mg to about 100 mg or 25 mg to about 50 mg.

The treatment regimen of an AI may vary depending on the particular subject involved, the density of the breast tissue and/or vascularity prior to treatment, the particular AI(s), and/or the intended result. The administration of one or more AI may be given as a single dose for one day or multiple days, or as multiple doses for one day or multiple days. Other pharmaceutical agents (e.g. hormone replacement therapy) may be administered before, concomitant with, or after administration of the AI. Administration of the AI(s) alone or in conjunction with other medications can start at least about 1 to about 2 days, 1 to about 3 days, 1 to about 5 days, 1 to about 7 days, 1 to about 10 days, 1 to about 14 days, 1 to about 20 days, 1 to about 30 days, 1 to about 60 days, 1 to about 120 days, 1 to about 180 days, 1 to about 200 days, 1 to about 210 days, 1 to 240 days, 1 week to 2 weeks, 1 week to 4 weeks, 1 week to 6 weeks, 1 week to 8 weeks, 1 week to 10 weeks, 1 week to 3 months, 1 week to 6 months, 1 week to 9 months, 1 week to 12 months, 1 month to 3 months, 1 month to 4 months, 1 month to 6 months, 1 month to 9 months, 1 month to 12 months, 4 months to 6 months, 4 months to 9 months, 6 months to 12 months, or 12 months prior to making an image of the breast.

The period of time that a subject is administered an AI(s) may vary depending on factors such as the particular subject involved, the density and/or vascularity of the breast tissue prior to treatment, the AI(s) used, and the desired degree of breast density and/or vascularity reduction. In aspects of the invention, a subject may be treated with an AI(s) prior to making an image of the subject's breast tissue continuously for about 1 to about 2 days, 1 to about 3 days, 1 to about 5 days, 1 to about 7 days, 1 to about 10 days, 1 to about 14 days, 1 to about 20 days, 1 to about 30 days, 1 to about 60 days, 1 to about 120 days, 1 to about 180 days, 1 to about 200 days, 1 to about 210 days, 1 to about 240 days, 1 week to 2 weeks, 1 week to 4 weeks, 1 week to 6 weeks, 1 week to 8 weeks, 1 week to 10 weeks, 1 week to 3 months, 1 week to 6 months, 1 week to 9 months, 1 week to 12 months, 1 month to 3 months, 1 month to 6 months, 1 month to 9 months, 1 month to 12 months, 6 months to 12 months, 12 months to 24 months, or 12 months to 48 months or longer, or periodically.

The invention further relates to a packaged formulation for reducing breast density in a subject prior to making an image of breast tissue of a subject. A packaged formulation of the invention comprises one or more AI packaged with instructions for using the one or more AI for reducing breast density and/or vascularity in a subject prior to making an image of the breast tissue of the subject. In an aspect of the invention, the packaged formulation comprises an effective amount of one or more AI. In an aspect of the invention, the AI in the packaged formulation may be in a form suitable for oral administration. In an aspect of the invention, the AI in the packaged formulation may be in a form suitable for systemic administration. In an aspect of the invention, the AI in the packaged formulation may be in a form suitable for subcutaneous or intravenous injection. In an aspect of the invention, the AI in the packaged formulation may be in a form suitable for continuous infusion. In an aspect of the invention, the AI in the packaged formulation is in a sustained release formulation.

A packaged formulation may be provided in a single dosage unit or multiple dosage units (i.e., tablets or capsules) having about 1 to about 1000 mg, 1 to about 500 mg, 1 to about 300 mg, 1 to about 250 mg, 1 to about 200 mg, 1 to about 150 mg, 1 to about 100 mg, 1 to about 75 mg, 1 to about 70 mg, 1 to about 60 mg, 1 to about 50 mg, 5 to about 60 mg, 5 to about 25 mg, 5 to about 20 mg, 10 to about 20 mg, 12 to about 25 mg, 15 to about 25 mg, or 15 to about 20 mg of an AI.

The methods and formulations of the invention may be used with or comprise any of a variety of aromatase inhibitors. A single aromatase inhibitor or a combination of aromatase inhibitors, in particular aromatase inhibitors with different half lives, may be used in the methods and formulations of the invention. An aromatase inhibitor may be selected that has a half-life of about 2 hours to about 1 day, 2 hours to about 3 days, 2 hours to about 4 days, 2 hours to about 2 days, 1 to about 5 days, 1 to about 3 days, 1 to about 2 days, or in particular 1 , 2, or 3 days.

Particular aromatase inhibitors that may be used in the methods of the invention include non-steroidal and reversible aromatase inhibitors or steroidal and non-reversible aromatase inhibitors. In aspects of the invention, an oral aromatase inhibitor, in particular a commercially available oral aromatase inhibitor is employed, including without limitation anastrozole (Arimidex™), letrozole (Femara™), vorozole and/or exemestane (Aromasin™). Preferably letrozole (Femara™), is employed in the present invention.

The invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes, and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters which can be changed or modified to yield essentially the same results. Example 1

Use ofAromatase Inhibitors to Decrease Breast Density in Menopausal Women on HRT The following methodology was employed in the study:

1. Patient selection Selection criteria:

• Postmenopausal women.

• Have been using hormonal replacement therapy for the management of menopausal symptoms.

• Have used an aromatase inhibitor (AI) for at least 4 months.

• Having regular annual mammograms done within the past 5-6 years or at least 2 mammograms dated just before (up to 1 year) and after (up to 1 year) the administration of the aromatase inhibitor. • No present or past history of breast cancer

Control mammograms were chosen from available mammograms of the same patients while they were on HRT alone prior to starting the AI. Additionally, mammograms of an age matched group of postmenopausal women using HRT alone were used. After obtaining the consents, all of the past and the present medical history were reviewed carefully and the data related to the study was collected and entered to coded electronic sheets.

2. Collection of Mammograms

Original mammogram films were recruited from different diagnostic imaging departments and clinics across the Greater Toronto Area after sending the required consents and ethics approval documents. Films were either picked by one of the investigators or received in mail. Most mammograms were returned within 3 weeks after finishing their digitization.

Two formats of mammograms were recruited: • Analog mammograms (Hard copies) : obtained by the conventional Film- based X-ray systems using an analog acquisition technique.

• Digital mammograms (CDs): obtained by Direct Digital Acquisition (DDA) and achieved through the picture archiving and communication systems (PACS) which are computer networks employed for the storage, retrieval, and distribution of digital images. PACS were available in recent mammograms post 2000 in Mount Sinai Hospital and Princess Margaret Hospital. The mammograms were provided on CDs with special software (eFilm (TM) Lite (TM) software Copyright ©1998-2005, Merge eMed) capable of manipulating the images in different ways and exporting them to other software. 3. Digitization /Scanning

The next step was to transform the mammograms available only in hard copies into digital forms that can be further analyzed by image analysis software. For this purpose two different pieces of equipment were used: a) Kodak LS85 film digitizer (Eastman Kodak Co., Rochester, NY). This is a high-resolution film digitizer which has a laser beam to illuminate scanned films and produce digital signals as accurate as 0.001 optical density through a logarithmic amplifier. The resolution and densities in the digital images produced by this digitizer are closely representative of the original image. The Kodak LS85 film digitizer Specifications are shown in Table 1. b) Agfa DuoScan Flatbed scanner : (Agfa-Gevaert Group- Mortsel, Belgium). This is a flatbed scanner which includes a transparency tray for transparencies up to 8.25" x 10". The scanner incorporates TwinPlate Scanning Technology to enable scanning of transparencies and reflective work on independent scan beds in a single integrated unit. Films are placed on a tray, which slides into the body of the scanner. A single carriage carrying light sources, optics, and a CCD sensor, traverses between the upper glass plate for reflective work and the transparency tray below. A rotation of a single mirror occurs upon switching from reflective to transparency scanning. The Agfa DuoScan Flatbed scanner specifications are shown in Table 2. Scanning of the films Preparation and placement of the films: In order to have an equal scanning environment necessary for comparison, each patient's mammograms were compared using the same scanner. Additionally each scanner's software was set to the same scanning parameters. Differences in the equipment or in any of these parameters would result in differences in the measurements of the breast density. Each patch of films was carefully arranged and labelled serially with a coded obscure name that could be interpreted only by the investigators into patient's ID, date of mammogram, laterality and view of the original film. Each film was then cleaned and placed inside the digitizer or inside the tray of the scanner. In the case of the Agfa scanner, proper application was important to make sure that no air is entrapped between the film and the glass tray which could create background noise that might affect the quality of the produced image. The Kodak digitizer needed at least 30 minutes to warm up before starting the process of scanning and it takes approximately 30 seconds to scan a single image while the Agfa scanner usually takes around 1 minute. After finishing the scanning process, the scanned images are saved by file names using a naming convention of the same labels tagged earlier on the X-rays; then each patient's files are stored in a single directory. Mammograms Scanning Software:

In order to acquire the image from the scanner, specific computer software is needed to be installed, through which previewing, managing, and setting the parameters of the scan can be done. There was specific software installed on the computer system linked to each of the scanners:

A. Lumisys Scan program is used with the Kodak LS85 film digitizer

With this equipment calibration steps are needed by the software at the start and end of each day of digitizing. It creates digital images in 2 formats: .iv2 and bmp. An iv2 is created at 260 microns per pixel and is 12 bits. For small films (18 x 24cm) an .iv2 is 1.2 MB and for large films (24 cm x 30 cm) an .iv2 is 2.05 MB. The bmp format is in 8 bits and is 0.6 MB for the small and 1.2 MB for the large films. The scanned image files are in RGB (red green blue) format which needs further transformation before computer assisted image analysis. The scan parameters were set as follows: 260 microns; X and Y offsets= 0; Averaging mode- none; 12 bits/pixel; MSB pixel format; IV2 format; User LUT= inverse; Make BMP on; and Average BMB off.

B. Fotolook FL 3.6 is used with Agfa DuoScan Scanner: This is used to preview, adjust and set up the profile of the scanned films. The images could be acquired directly in grayscale mode and the images could be cropped in a preview mode if desired. After scanning the image, it was saved into the "Tagged Image File Format" (TIFF) which is suitable for further image analysis. Produced file sizes were approximately 78MB. The scan parameters were set as follows: Original: Transparent; Size: Max. area; Mode : Gray Scale; Input: 800ppi; Scale to: 100%; 16 bit; Range: Automatic; Tone curve: None; Sharpness: high; Descreen: None; Flavor: none; and w^ point: none.

4. Conventional qualitative and semi quantitative analysis of the mammograms- Role of the Radiologist

A single experienced radiologist in breast imaging at the Marvelle Koffler Breast Centre at Mount Sinai Hospital and the breast imaging department of Princess Margaret Hospital were responsible for the analysis of the mammograms by visual inspection. The radiologist reviewed the mammograms after their digitization and not in hard copies. The radiologist was totally blinded to any information related to the patients, the dates of the mammograms or if a specific film was obtained before or after the investigated protocol. The file names of the images appearing on the screen were all in an obscure code that could not be interpreted by the radiologist as identifying data. Films were also shown by the investigator to the radiologist randomly without specific order. The radiologist analyzed each image separately and comments were recorded by the investigator. In order to make for intraobserver reliability, randomly taken samples of the films were reblinded and shown again to a breast-imaging specialist.

Two commonly used scoring methods were applied:

A. The Breast Imaging Reporting and Data System (BI-RADS): BI-RADS is a qualitative method developed by the American College of Radiology

(ACR), which provides a standard classification for mammographic breast density estimations and facilitates the reporting of patients' results and the communication among health providers. Table 3 shows the interpretation of BI-RADS scores

B. Semi-quantitative analysis using visual measuring of percent breast density: Five categories were assigned for measuring breast density: <10%; 10-25%; 26-

50%; 51-75%; and 76-100%.

5. Quantitative Computer Assisted Image analysis: In total, there were 3 different sources of the images that were eventually collected as shown in Figure 1. There were well visible differences in the quality and resolution of the produced images among the three methods. The highest quality being for digitally acquired films followed by films scanned by the Kodak LS85 digitizer and the least quality was produced by the Agfa Duoscan flatbed scanner. Additionally, a previous study found that breast density was significantly lower in mammograms originally acquired in digital format compared to those digitized after being obtained by the conventional film screen (Harvey, J. 2004). However, as mentioned above each patient's group of films assigned for comparison were screened and scanned using the same equipment. Though all the available views were collected of both breasts (right and left cranio caudal (RCC, LCC), and right and left mediolateral oblique (RMLO, LMLO), but for the scanning and comparison purposes the RCC projection was mostly used, based on previous data of the high symmetry existing between different projections (Bying JW et al, 1996). One mammogram before the start of the aromatase inhibitor treatment and the most recent mammogram during the administration of the drug were compared.

Two image analysis tools were used:

L ImageQuant: ImageQuant Version 5 (Copyright ©1998 Molecular Dynamics Sunnyvale,CΛ )

This is a multi-purpose program commonly used in biomedical labs. It offers different modules for various types of samples. Every module has a simple wizard interface. Different editing tools are included in the program (Figure 2). In the field of breast imaging, this program has been used for the purpose of analyzing mammographic breast density and it showed comparable results to both the visual analysis and the commonly used Cumulus program (Cumulus 108 software, University of Toronto) (Palomares, MR et al, 2006). The ImageQuant software calculates the area using the number of pixels for the outlined region. Most importantly it takes in consideration the volume averaging from projecting the image of a three-dimensional breast onto a two-dimensional film, and therefore it calculates a volume estimate as well. ImageQuant then assigns each pixel a numerical value according to the absorbance of the image at this spot. Eventually, it presents the summation of all those numerical values in what is termed : the integrated pixel intensity (IPI) (Palomares, MR et al, 2006). Steps of Analysis: For optimal analysis, images should be in the TIFF format 16-bit gray scale. Therefore digitally acquired images or images scanned by the Kodak LS85 digitizer were transformed from the RGB(red green blue) format into gray scale 16 bit by using Adobe® Photoshop® CS2 program. Images scanned by Agfa were already saved in this format. The two mammograms assigned for comparison were opened simultaneously in the program and the gray scale was adjusted to clearly visualize the breast edge. The whole breast was outlined using the mouse. Different tools are available for region selection; the polygonal tool was chosen which provided better control and more accurate outline of the breast edge. Chest wall structures, when present, were excluded from the outline. Using the copy- paste tool, the polygon drawn above is pasted over the breast in the second image. When there is a discrepancy between the sizes of the breast visualized in the two films, then the smaller breast is outlined first and copied and pasted to the larger breast starting from the nipple side to avoid biased measurements based on patient positioning which might happen during the original film screen. (Figure 3). Then, in order to avoid the effect of the background noise on the measured (IPI), background correction tools available in ImageQuant were used. There are different types of background correction methods that subtract the calculated background value from the IPI to result in the final volume estimate for the outlined area. The "none" correction option was used when the background value is zero as in the digitally acquired films. "Local Average" option value was selected when the background is heterogeneous allover the film surface, and this option calculates a background value which is equal to the average of all the points beneath the outline. Background correction can be also done using an Object Average option, which should be best used when the background noise is homogenous. In most films, heterogeneous backgrounds were seen, so in addition to the "Local Average", the "Object Average" tool was adjusted by drawing a box that extends across the film to account for the different densities in the background away from the breast area and the box was automatically pasted over the second image (Figure 3). The gray scale was readjusted again to enhance contrast between fatty and glandular tissue, then the total area of glandular tissue and the densest glandular region were outlined (Figure 4). The "IPI "was then calculated for all selected regions along with a large number of other useful measurements using the Report Volume function. The output could be exported to the Excel program (Microsoft Office Excel 2003) and saved in .xl excel file format. ii. Image J : (National Institute of Health NIH)

Image J is a free public domain Java image processing program offered by the NIH (NIH, Image J). It can be used, either as an online applet or can be downloaded to any computer. This system should have Java 1.4 (Sun Microsystems, Inc.) or later versions installed. It also has more than 300 plugins that can be downloaded and added to enhance its function. Using image J, the average intensities of the pixels inside an outlined area similar to ImageQuant were measured; however there was no volume measurement feature. On the other hand, Image J provided a more visual representation of the breast density by its "Surface Plot" function (Figure 5). Interactive surface plot is also available as a plugin (Figure 6). Identifying patient information and other extraneous marks should be removed by cropping the image before running the surface plot function to avoid confusion with the breast tissue. Grayscale images can be also displayed in colors through the "lookup tables" tool which allows for a broad selection of colors and patterns that can be assigned for each of 256 possible displayed pixel values (Figure 7). 6. Data Recording and Statistical Analysis

ImageQuant data output was exported to Excel software (Microsoft Office Excel 2003). The statistical tests were then performed using SPSS 14.00 for Windows (Release 14.01, SPSS Inc., Chicago, IL, USA). Both the paired samples t-test and the non parametric Wilcoxon's test were used to analyze related continuous variables and the independent student's Mest and the non-parametric Mann- Whitney Latest were used to analyze continuous independent variables. P < 0.05 was considered statistically significant. Results:

The results are shown in Figures 8 to 11 and Tables 4, 5 and 6. The results show a significant reduction in breast density in women who have received an aromatase inhibitor. Thus, aromatase inhibitors, preferably in large doses, can be used to decrease breast density prior to mammogram to improve accuracy of the test. Example 2

A study will be performed to determine if giving a high dose of aromatase inhibitors prior to MRI could, through the marked suppression of local estrogenic milieu, improve the MRI specificity by reducing the rapid and marked uptake of highly vascular benign breast tissues and improve the MRI sensitivity by suppressing the background signal intensity of normal breast tissue through reducing breast density. The main difference between MRI and other imaging techniques is the accompanying intravenous injection of a contrast material, commonly Gadolinium, which is a non radioactive material specifically up-taken by highly vascular tissues, especially cancer, leading to enhancement or brightness of this particular tissue. The malignant tissue is typically able to enhance more markedly and quickly than normal tissue or benign lesions possibly due to an abnormal and extremely high vascularity. Though the sensitivity of MRI is very high, up to 90%, its specificity is much lower which means more benign tissues could similarly enhance and cause false positive results resulting in low positive predictive values (Sardanelli, 2004;. Piccoli, 1997). (Figure 13 shows benign background enhancement in MRI and its Image J interactive surface plotting.) Also, malignant MRI enhancement could appear as a non-mass like pattern in a considerable number of breast cancer patients representing another MRI diagnostic difficulty (Schrading, 2008).

The primary objectives of this study are to determine the effectiveness of the aromatase inhibitor Letrozole in increasing the sensitivity and specificity of MRI in healthy postmenopausal women with diagnostic difficulties. Secondary obj ectives are to determine the tolerability and side effects of this intervention at different doses.

The following are the primary and secondary outcome measures: Primary Outcome Measures (Efficacy Analysis):

1. Quantitative percentage change in MRI signal intensity between the pre and the post treatment MRI.

2. Objective analysis of breast density, volume and composition using breast MRI.

3. Assessment of confidence of radiologist to conclude a diagnosis Secondary outcome Measures (Safety Analysis):

1. Subjective responses of study subjects about side effects that could be related to aromatase inhibition.

2. Blood pressure

The following describes the study design and duration: Study Type: Interventional

Study Design: Pilot prospective study with a clinically similar retrospective cohort as control group.

Eligibility: Age eligibility: 35 years old and over. Gender eligibility. Female. Target population: Healthy postmenopausal who have had one or repeated MRIs for diagnostic difficulties.

Brief Outline:

Study subjects will be enrolled in one of three arms:

• Arm I: Patients receive oral letrozole 5 mg daily for 3 days. • Arm II: Patients receive oral letrozole 12.5 mg daily for 3 days.

• Arm III: Patients receive oral letrozole 20 mg daily for 3 days. Total study duration: 6 months

Post treatment period: One month

Sample Size: A total of 60 women with 20 women in each arm will be enrolled for this study. Another 60 retrospective cases of MRI will serve as a control group. Inclusion Criteria

• Postmenopausal, defined by any of the following criteria:

> Cessation of menstrual bleeding of duration of 12 months or more

> Prior hysterectomy and bilateral salpingo-oophorectomy. > > 55 years of age with a prior hysterectomy with or without oophorectomy.

> < 55 years of age with a prior hysterectomy without oophorectomy OR the status of the ovaries is unknown and follicle-stimulating hormone (FSH) level is in the postmenopausal range

• Have had a previous MRI within the past month or been already scheduled for an MRI due to diagnostic difficulty or high risk screening population.

Exclusion Criteria

• Present or past diagnosis of breast cancer.

• Patients with any implant including pacemakers, vagus nerve stimulators, implantable cardioverter-defibrillators (ICD), loop recorders, insulin pumps, cochlear implants, deep brain stimulators, surgical prostheses, aneurysm clips or any other implant.

• Present or past history of renal impairment (Serum creatinine >2.0 mg/dL) or renal disease.

• Osteoporosis: defined as a bone mineral density T-score > -2.5 on dual-energy x- ray absorptiometry. History of pathological or fragility fractures.

• Mental health status resulting in cognitive or emotional impairment that would preclude study participation. Any medical condition that would jeopardize the patient or the integrity of the data obtained including advanced cardiac, renal, liver diseases, blood, metabolic severe dementia, advanced cancers in any body site, etc.

• Concurrent or prior use within the past 3 months or of gonadotrophic releasing hormone analogues, selective estrogen receptor modulators, phytoestrogens, other aromatase inhibitors, androgens and prolactin inhibitors.

Drug Formulation: Femara ™ Novartis Pharmaceuticals Canada Inc., Dorval, QC). Oral Tablets 2.5 mg.

Dosage Regimen: 5 mg, 12.5 mg and 20 mg once daily for 3 successive days according to the study arm. Study subjects will be randomized for each of these three arms. Concomitant Medications

The following drugs are disallowed 3 months before and during the study period:

Selective estrogen receptor modulators (SERMs); Gonadotrophin releasing hormone analogues (GnRh agonists and antagonists); Phytoestrogens; Other aromatase inhibitors

(steroidal and non steroidal); Androgens; Prolactin inhibitors and Other investigational drugs.

The following outcome analysis methods will be used:

1. Quantitative percentage changes in signal intensity between the pre and the post treatment MRI. MRIs are obtained and archived through the Picture Archiving and Communication Systems (PACS). The MRIs will be burned on CDs with a DICOM viewer (eFilm (TM) Lite (TM) software Copyright ©1998-2005, Merge eMed, Milwaukee, WI USA) that allowed manipulation of the images and exporting them to other computer programs. Transverse sections will be used for the purpose of this study. The level of the nipple will be taken as a landmark to compare the corresponding sections in both the pre and post treatment films. The MRI series (Tl) will be used for the evaluation and comparison of enhancement in all study subjects and controls. Subtraction views will be measured for signal intensities (SI) (Figure 12) (Medscape, 2001). Both the right and left breast will be analyzed. Image analysis software including Image J (http://rsb.info.nih.gov/ij/) and ImageQuant (IQ) Version 5 (Copyright ©1998 Molecular Dynamics Sunnyvale, CA) will be used to analyze signal intensity (SI) in the compared images. In both programs the integrated pixel intensity will be calculated as described above. Enhancement after gadolinium dye injection will be calculated based on the following equation (Rieber, 1997): Enhancement = SI postcontrast - SI precontrast X 100

SI pre-contrast

SI postcontrast will be determined at 3 time points after Gadolinuim injection.

2. Objective assessment of breast density and vascularity by MRI. Transverse sections will be used to calculate the integrated pixel intensity (IPI) of the overall change of breast density and vascularity using the above described softwares in both the treatment and the control groups. 3. Assessment of confidence of radiologist to conclude a diagnosis. The objective assessment of time taken by the radiologist to conclude a diagnosis will be determined by another blinded investigator and compared between the treatment group and the retrospective control group. The subjective assessment of the clarity of image visualization and identification of a benign versus malignant lesions will be determined by assigning a score for clarity of image and easiness of identifying pattern of enhancement (benign versus malignant) pre and post treatment and in the control MRIs. The same radiologist will assign the scores for all cases.

4. Assessment for the retrospective control group: A clinically similar retrospective cohort will be chosen as a control group. Similarity is based on age, body mass index, hormone therapy status and breast cancer risk. Breast cancer risk is based on familial and genetic risk factors as well as Gail risk score. The same outcome measures mentioned above will be applied for the retrospective cohort of MRI. Two successive MRIs with 4 months or less apart will be compared to see if there is a difference between the change of MRI pattern of enhancement and density with time and without an intervention being applied.

Data Analysis. Statistical Analysis will include the analysis of a subject's base line clinical features, risk indicators as well as the baseline and after treatment endpoints. The non- parametric Mann-Whitney U-test and repeated measures ANOVA will be used to analyze independent and related continuous variables respectively, Wilcoxon's signed-rank test to analyze related continuous variables, the chi-square (χ2) test to analyze categorical variables and Spearman's test in measuring bivariate correlations. Table 1

Table 2

Table 3

Table 4

Age: Control Group vs Treatment Group

Table 5

Control Group Integrated Pixel Intensity (total volume)

Table 6

AI Treatment Group Integrated Pixel Intensity (total volume)

References:

1. Kodak, http://www. kodak. com/eknec/documents/59/0900688a802f7759/EN LS85. vdf. 2006.

2. Bassignani, M.J., A.P. LoRusso, and J.A. Harvey, Digitizing and Consolidating Mammograms and Other Images for Teaching Applications. Academic Radiology, 2006. 13(6): p. 774-781.

3. AGFA, FotoLook FL 3.6 http://www. agfa. com/digicam scanner drivers/ scanner/ duoscan/index. html.

4. Eberl, M. M., et al., BI-RADS Classification for Management of Abnormal Mammograms. J Am Board Fam Med, 2006. 19(2): p. 161-164.

5. Palomares, M.R., et al., Mammography Density Correlation with Gail Model Breast Cancer Risk Estimates and Component Risk Factors. Cancer Epidemiol Biomarkers Prev, 2006. 15(7): p. 1324-1330.

6. Harvey, J., Quantitative assessment of percent breast density: analog versus digital acquisition. Technol Cancer Res Treat. , 2004. 3((6)): p. 611-6.

7. Byng JW, B.N., Little L, Lockwood G, Fishell E, Jong RA, Yaffe MJ, Symmetry of projection in the quantitative analysis of mammographic images. Eur J Cancer Prev., 1996. 5(5): p. 319-27.

8. NIH, Image J. Image Processing And Analysis In Java http://rsb. info, nih. gov/ii /index, html, W. Rasband., Editor. National Institute of Health.

9. Bedrosian l, MickR, Orel SG, SchnallM, Reynolds C, Spitz FR, CallansLS, Buzby GP, Rosato EF, Fraker DL, Czerniecki BJ 2003 Changes in the surgical management of patients with breast carcinoma based on preoperative magnetic resonance imaging. Cancer 98:468-473

10. Hagen AI, KvistadKA, Maehle L, Holmen MM, Aase H, Styr B, Vabo A, ApoldJ, Skaane P, Moller P 2007 Sensitivity of MRI versus conventional screening in the diagnosis of BRCA-associated breast cancer in a national prospective series.

Breast 16:367-374

11. Komenaka IK, DitkoffBA, Joseph KA, Russo D, Gorroochurn P, Ward M Horowitz E, El-Tamer MB, Schnabel FR 2004 The Development of Interval Breast Malignancies in Patients with BRCA Mutations. Cancer 100:2079-2083 12. Medscape 2001 http://www.medscape.com/content/2001/00/40/90/409021/art- cc0805.01.figl.jpg. Medscape 13. Rieber A, ZeitlerH, Rosenthal H, GorichJ, Kreienberg R, Brambs HJ, TomczakR

1997 MRI of breast cancer: influence of chemotherapy on sensitivity. Br J Radiol

70:452-458 14. Sardanelli F, Giuseppetti GM, Panizza P, Bazzocchi M, Fausto A, Simonetti G,

Lattanzio V, Del Maschio A 2004 Sensitivity of MRI versus mammography for detecting foci of multifocal, multicentric breast cancer in Fatty and dense breasts using the whole-breast pathologic examination as a gold standard. AJR Am J

Roentgenol 183:1149-1157 15. Piccoli CW 1997 Contrast-enhanced breast MRI: factors affecting sensitivity and specificity. Eur Radiol 7 Suppl 5:281-288 16. SchradingS, Kuhl CK 2008 Mammographic, US, and MR imaging phenotypes of familial breast cancer. Radiology 246:58-70

The recitation of numerical ranges by endpoints herein includes all numbers and fractions subsumed within that range (e.g. 1 to 5 includes 1 , 1.5, 2, 2.75, 3, 3.90, 4, and 5). It is also to be understood that all numbers and fractions thereof are presumed to be modified by the term "about." Further, it is to be understood that "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to an "aromastase inhibitor" includes a mixture of two or more aromatase inhibitors The term "about" means plus or minus 0.1 to 50%, 5-50%, or 10-40%, preferably 10-20%, more preferably 10% or 15%, of the number to which reference is being made. The present invention is not to be limited in scope by the specific embodiments described herein, since such embodiments are intended as but single illustrations of one aspect of the invention and any functionally equivalent embodiments are within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

All publications, patents and patent applications referred to herein are incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. The citation of any reference herein is not an admission that such reference is available as prior art to the instant invention.

Claims

WHAT IS CLAIMED IS:

1. A method for detecting an abnormality in breast tissue in a subject comprising administering to the subject an effective amount of one or more aromatase inhibitor to reduce the density and/or vascularity of the breast tissue in the subject and making an image of the breast tissue such that the abnormality in the breast is capable of being, or is detected.

2. A method according to claim 1 wherein the abnormality is a mass, a cyst, a microcalcification, a fibrous finding, an architectural distortion, and/or other abnormal findings related with benign or malignant abnormalities.

3. A method according to claim 1 wherein the abnormality is a benign or malignant tumor.

4. A method according to claim 1 wherein the image is made of breast tissue using film mammography, digital mammography, ultrasonography, thermography, computed tomography, positive emission tomography, transillumination, magnetic resonance imaging (MRI), or radionucleotide imaging.

5. A method according to claim 4 wherein the image is made of breast tissue using mammography.

6. A method according to claim 4 wherein the image is made of breast tissue using magnetic resonance imaging.

7. A method for diagnosing breast cancer in a subject comprising administering an effective amount of an aromatase inhibitor to the subject and subjecting the subject to a mammogram procedure.

8. A method of preparing a female patient for a mammogram procedure comprising the step of, prior to performing the procedure, administering to the patient one or more doses of at least one aromatase inhibitor, either alone or in conjunction with another pharmaceutical agent, in an amount effective to reduce breast density in the subject.

9. A method for diagnosing breast cancer in a subject comprising administering an effective amount of an aromatase inhibitor to the subject and subjecting the subject to a breast MRI.

10. A method of preparing a female patient for a breast MRI comprising the step of, prior to performing the MRI, administering to the patient one or more doses of at least one aromatase inhibitor, either alone or in conjunction with another pharmaceutical agent, in an amount effective to reduce vascularity in the subject.

11. A method according to any preceding claim wherein the aromatase inhibitor has a steroidal or non-steroidal chemical structure.

12. A method according to any preceding claim wherein the aromatase inhibitor is selected from non steroidal and reversible aromatase inhibitors.

13. A method according to any preceding claim wherein the aromatase inhibitor is an inhibitor selected from the group consisting of anastrozole, letrozole, vorozole and exemestane.

14. A method according to any preceding claim wherein the aromatase inhibitor is letrozole administered in a daily dose of from about 2.5 mg to about 60 mg.

15. A method according to any preceding claim wherein the aromatase inhibitor is anastrozole administered in a daily dose of from about 1 mg to about 100 mg.

16. A method according to any preceding claim wherein the aromatase inhibitor is vorozole administered in a daily dose of from about 4 mg to about 100 mg.

17. A method according to any preceding claim wherein the aromatase inhibitor is exemestane administered in a daily dose of from about 25 mg to about 500 mg.

18. A method according to any preceding claim wherein the aromatase inhibitor is administered orally.

19. A packaged formulation for reducing breast density and/or vascularity in a subject prior to making an image of the subject's breast tissue comprising one or more aromatase inhibitor packaged with instructions for using the aromatase inhibitor for reducing breast density and/or vascularity in the subject prior to making an image of the subject's breast tissue.

20. Use of one or more aromatase inhibitor either alone or in combination with other pharmaceutical agents for reducing breast density and/or vascularity in a subject before making an image of the subject's breast tissue.

21. Use of an aromatase inhibitor for the manufacture of a medicament for use in combination with mammography or MRI in a subject.