Case IPR20 1 7-00 900
`
`Declaration of Dorraya El-Ashry, Ph.D. Under 37 C.F.R. § 1.68 in Support of
`Petition for Inter Partes Review of U. S. Patent No. 8,329,680
`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`
`BEFORE THE PATENT TRIAL AND APPEAL BOARD
`
`INNOPHARMA LICENSING, LLC,
`Petitioner
`
`V.
`
`ASTRAZENECA AB,
`Patent Owner
`
`Case IPR2017-00900
`
`Patent No. 8,329,680
`
`
`DECLARATION OF DORRAYA EL-ASI-IRY Ph.D. UNDER 37 C.F.R.
`
`§ 1.68 IN SUPPORT OF PETITION FOR INTER PARTES REVIEW OF U. S.
`PATENT N0. 8,329,680
`
`Mail Stop: Patent Board
`Patent Trial and Appeal Board
`United States Patent and Trademark Office
`
`PO. Box 1450
`
`Alexandria, VA 22313-1450
`
`InnoPharma Exhibit 1014.0001
`
`

`

`Case IPR2017—00900
`
`Declaration of Dorraya El-Ashry, PhD. Under 37 C.F.R. § 1.68 in Support of
`Petition for Inter Partes Review of U.3. Patent No. 8,329,680
`
`TABLE OF CONTENTS
`
`I.
`
`INTRODUCTION ........................................................................................ .. 1
`
`II. BACKGROUND AND QUALIFICATIONS ............................................... .. 2
`
`III. MATERIALS CONSIDERED FOR THIS DECLARATION ...................... .. 6
`
`IV. SUIVIMARY OF OPINIONS ........................................................................ .. 6
`
`V. LEVEL OF ORDINARY SKILL IN THE PERTINENT ART .................... .
`
`. ll
`
`VI. OVERVIEW OF TYPES OF BREAST CANCER AND THE AVAILABLE
`
`THERAPIES AS OF 2000 .......................................................................... ..12
`
`A. Hormone-Dependent Breast Cancer .................................................. .
`
`. 12
`
`(1) SERMs ........................................................................................ ..13
`
`(2) AIs .............................................................................................. ..14
`
`(3) ERDs ........................................................................................... ..15
`
`B. Hormone-Independent Breast Cancer ................................................ ..15
`
`VII. OVERVIEW OF THE MCLESKEY REFERENCE .................................... .
`
`. 17
`
`A.
`
`Summary of the Testing Conducted in the McLeskey Reference ....... ..17
`
`B. Results Reported in the McLeskey Reference ................................... ..19
`
`C. Conclusions That A Skilled Researcher Would Draw From the
`
`McLeskey Reference ......................................................................... .22
`
`VIIIRESPONSE TO THE MCLESKEY DECLARATION ............................... .23
`
`A. Fulvestrant Was Not a Treatment Failure .......................................... .23
`
`B. A Skilled Researcher Would Not Have Concluded That Preformulated
`
`Fulvestrant Could Only Be Administered to Animals ........................ .25
`
`C. A Skilled Researcher Would Have Considered the McLeskey Reference
`
`ii
`
`InnoPharma Exhibit 1014.0002
`
`

`

`Case IPR20 1 7-00 900
`
`Declaration of Dorraya El-Ashry, PhD. Under 37 C.F.R. § 1.68 in Support of
`Petition for Inter Partes Review of U. S. Patent No. 8,329,680
`
`Relevant and Helpful ......................................................................... ..27
`
`IX. CONCLUSION ........................................................................................... . .29
`
`iii
`
`InnoPharma Exhibit 1014.0003
`
`

`

`Case IPR20 1 7-00 900
`
`Declaration of Dorraya El-Ashry, Ph.D. Under 37 C.F.R. § 1.68 in Support of
`Petition for Inter Partes Review of U. S. Patent No. 8,329,680
`
`I, Dorraya El—Ashry, PhD. hereby declare as follows:
`
`I.
`
`INTRODUCTION
`
`1.
`
`I have been retained as an expert witness on behalf of lnnoPharma
`
`Licensing, LLC (“lnnoPharma”) for the above-captioned Petition for Inter Partes
`
`Review (“IPR”) of US. Patent No. 8,329,680 (“the ‘680 patent”).
`
`I am being
`
`compensated for my time in connection with this lPR at my standard consulting
`
`rate of $500 per hour. My compensation is in no way dependent on the outcome of
`
`this matter.
`
`2.
`
`I have been asked to provide my opinions regarding the article
`
`Tamoxzfen-resistant Fibroblast Growth Factor-transfected MCF-7 Cells Are
`
`Cross-Resistant in Vivo to the Antiestrogen [Cl 182,780 and Two Aromatase
`
`Inhibitors,
`
`4 Clinical Cancer Research 697-711
`
`(1998)
`
`(“the McLeskey
`
`Reference”).
`
`1 am an author on the McLeskey Reference, and was the principal
`
`estrogen receptor expert on the project. The remaining authors of the McLeskey
`
`reference—including Drs. McLeskey and Kern—were growth factor experts, not
`
`estrogen receptor experts.
`
`3.
`
`I have also been asked to respond to certain opinions set forth in the
`
`Declaration of Sandra McLeskey (“the McLeskey Declaration”), which I
`
`understand that AstraZeneca AB (“AstraZeneca”) attached as Exhibit 2043 to its
`
`InnoPharma Exhibit 1014.0004
`
`

`

`Patent Owner Response in IPR2016-01316, -01324, -01325, and -01326.
`
`4.
`
`In preparing this Declaration, I have reviewed the ‘680 patent, the
`
`McLeskey Reference,
`
`the McLeskey Declaration,
`
`and the
`
`article
`
`titled
`
`Pharmacokinetics, Pharmacological and Anti-tumor Effects of the Specific Anti-
`
`Oestrogen ICI 182780 in Women with Advanced Breast Cancer, BRITISH J. OF
`
`CANCER, 74, p. 300-308 (1996) (“the Howell Reference”).
`
`5.
`
`In forming the opinions expressed in this Declaration, I relied upon
`
`my education and experience in the relevant field of the art, and have considered
`
`the viewpoint of a person having ordinary skill in the relevant art as of 2000.
`
`II.
`
`BACKGROUND AND QUALIFICATIONS
`
`6.
`
`I received my BA.
`
`in Molecular Biology in 1984 from Vanderbilt
`
`University, and my PhD. in Experimental Pathology in 1989 from the University
`
`of Colorado Health Sciences Center. My dissertation focused on the cellular and
`
`molecular properties of progesterone receptors in breast cancer.
`
`7.
`
`After completing my Ph.D., I began a Post-Doctoral Fellowship at the
`
`Georgetown Lombardi Cancer Center in 1994 in Washington, DC. My research
`
`area was estrogen regulation of TGF-oc in breast cancer. While at the Lombardi
`
`Cancer Center, I helped draft the McLeskey Reference along with my co-authors,
`
`including Dr. McLeskey and Dr. Kern.
`
`8.
`
`I was promoted to Research Assistant Professor at the Lombardi
`
`InnoPharma Exhibit 1014.0005
`
`

`

`Cancer Center in 1996 and was subsequently appointed to the Department of
`
`Oncology in 1999, where I was promoted again in 2000 to Assistant Professor.
`
`I
`
`remained in this position until 2001, when I moved to the University of Michigan
`
`Health System and was appointed Assistant Professor in the Department of
`
`Medicine, Division of Hematology/Oncology. For the 7 years I was at
`
`the
`
`University of Michigan, I was the Principle Investigator of a breast cancer research
`
`lab — the research focus of my was the generation of the estrogen receptor (ER)-
`
`negative phenotype in breast cancer by altered grth factor signaling pathways,
`
`and mechanisms for re-expression of ER and thereby potential restoration of anti-
`
`estrogen responses. At U of M I also trained graduate students and post-doctoral
`
`fellows,
`
`taught, served on committees and was a member of the Institutional
`
`Review Board (IRB).
`
`9.
`
`In 2008,
`
`I moved to the University of Miami Miller School of
`
`Medicine, Sylvester Comprehensive Cancer Center”s Braman Family Breast
`
`Cancer Institute, where I was appointed Associate Professor in the Department of
`
`Medicine, Division of Hematology/Oncology. Here at W, I am the PI of a breast
`
`cancer research lab where the focus of our research continued to be mechanisms of
`
`ER repression and its re-expression with restoration of anti-estrogen responses. We
`
`moved this work into the clinic where we wanted to determine the frequency of ER
`
`re-expression in ER-negative cancers via an inhibitor of a growth factor signaling
`
`InnoPharma Exhibit 1014.0006
`
`

`

`molecule. More recently, our focus has moved onto the tumor microenvironment
`
`(Th/IE) which is the normal cells surrounding a cancer that have been usurped by
`
`the cancer to support the cancer’s behavior, and we focus on a particular cell type
`
`in of the TlVlE, cancer associated fibroblasts or CAFs. We have studied the role of
`
`CAFs in repressing ER expression in the cancer cell, as well as specific differences
`
`in CAFs from ER+, luminal breast cancers compared with CAFs from ER-, basal
`
`breast cancers, and have most recently demonstrated that CAFS circulate in the
`
`blood of cancer patients along with circulating tumor cells (CTCs).
`
`I
`
`train
`
`graduate students, post-doctoral and clinical fellows, sit on several committees, and
`
`am ad—hoc member of the IRB.
`
`10.
`
`I have spent my entire career studying breast cancer, with a specific
`
`focus on the role of the estrogen receptor in breast cancer, and mechanisms for
`
`restoration of anti-estrogen therapy responses.
`
`In particular, I have authored more
`
`than 35 peer-reviewed publications, more than 50 abstracts (presented at National
`
`and International meetings), and review articles, many of which specifically
`
`focused on the role of the estrogen receptor in breast cancer. See, e. g, Holloway et
`
`al., A Cytoplasmic Substrate ofMAPK is Responsible for ERa Down-Regulation in
`
`Breast Cancer Cells: Role of NFKB, 18 Molecular Endocrinology 1396-1410
`
`(2004); Creighton et al., Activation ofMAPK in ERa-positive breast cancer cells in
`
`vitro induces an in vivo molecular phenotype of ERa-negative human breast
`
`InnoPharma Exhibit 1014.0007
`
`

`

`tumors, 66 Cancer Research 3903-11 (2006)', Shah et al., Hierarchical Paracrine
`
`Interaction of Breast Cancer Associated Fibroblasts with Cancer Cells via
`
`hMAPK-microRNAs to Drive ER-Negative Breast Cancer Phenotype, 16 Cancer
`
`Biology & Therapy 1671-81 (2015).
`
`11.
`
`l have also served as the Principal or Co-lnvestigator in close to 20
`
`grants for research studies focused on studying the role of the estrogen receptor
`
`(ER) in the development and treatment of breast cancer. See, e. g., Reversing the
`
`ER-Negative Phenotype in Breast Cancer (2001-2002), SWI/SNE: Mediator of the
`
`ER +/PR—, Tamoxifen-Resistant Phenotype in Breast Cancer? (2004), Mechanisms
`
`Underlying the Acquisition of the ER-Negative Phenotype in Breast Cancer (2001-
`
`2004), SWI/SNE.‘ Mediator of the ER+/PR-, Tamoxifen-Resistant Phenotype?
`
`(2004-2006), Restoring ER Expression and Anti-Estrogen Response to ER-
`
`Negative Breast Cancers (2006-2008), Mechanisms Underlying ER-Negative
`
`Breast Cancer (2006-2010), ER Expression and Anti-Estrogen Response in ER-
`
`Negative Breast Cancer (2010-2012).
`
`12.
`
`I am currently a member of the Editorial Board of PloS ONE (since
`
`2008) and Cancer Biology and Therapy (since 2001). I previously served as a
`
`Scientific Editor on the Senior Editorial Board of Endocrine-Related Cancer
`
`(2011-2016).
`
`I have also served as a reviewer for a number of peer-reviewed
`
`journals,
`
`including Molecular Endocrinology, Endocrinology, Journal of the
`
`InnoPharma Exhibit 1014.0008
`
`

`

`National Cancer Institute, Oncogene, Breast Cancer Research and Treatment,
`
`Breast Cancer Research, Cancer Research, Clinical Cancer Research, Cancer
`
`Prevention Research, Molecular Cancer Research, Neoplasia, Journal of
`
`Biological Chemistry, Journal of Cellular Physiology, and Journal of Steroid
`
`Biochemistry and Molecular Biology.
`
`13.
`
`I am also a member of numerous professional societies.
`
`In particular,
`
`I have also played an active role in the American Association for Cancer Research,
`
`serving as the chair or co-chair for numerous meetings,
`
`including meetings
`
`focusing on steroid and nuclear receptor/growth factor signaling pathways, as well
`
`as endocrinology more generally.
`
`I also actively participate in the Endocrine
`
`Society, including serving as a chair at annual meetings.
`
`14. My academic and research background, qualifications, and a complete
`
`list of my publications over my career are set forth in my curriculum vitae, which
`
`is attached as Exhibit A to this declaration.
`
`I have not testified as an expert at trial
`
`or deposition over the past four years.
`
`III. MATERIALS CONSIDERED FOR THIS DECLARATION
`
`15.
`
`In addition to my general knowledge, education, and experience, I
`
`considered the materials listed in Exhibit B in forming my opinions.
`
`IV.
`
`SUMMARY OF OPINIONS
`
`16.
`
`Based on my review of the McLeskey Declaration and the McLeskey
`
`InnoPharma Exhibit 1014.0009
`
`

`

`Reference,
`
`the other materials I have considered, and my knowledge and
`
`experience, my opinions are as follows:
`
`0 Dr. McLeskey and AstraZeneca misinterpret the McLeskey Reference when
`
`they characterize its use of fulvestrant as a “treatment failure.”
`
`0 As I explain below, fulvestrant worked exactly as it was intended in the
`
`McLeskey Reference—by inhibiting the estrogen receptor. In fact, as Dr.
`
`McLeskey explains,
`
`fulvestrant was used as a control
`
`in the study
`
`because it was well-known in the art to inhibit the estrogen receptor.
`
`McLeskey Decl. ll 5 (fulvestrant was used to “make sure that the estrogen
`
`receptor
`
`(ER) was not activated by small amounts of estrogen
`
`synthesized by the mouse’s liver and adrenal glands—with the goal being
`
`to determine if the activity of FGF (rather than estrogen) could drive
`
`tumor growth in tamoxifen-resistant breast cancer cells”).
`
`0
`
`In the control
`
`(“NIL-20”) experiments reported in the McLeskey
`
`Reference, fulvestrant successfully inhibited growth. Exhibit 1008 at
`
`Figure 4, Figure 5.
`
`0 We discovered that the fibroblast growth factor (“FGF”) transfected cell
`
`line that we used in the study was resistant to tamoxifen (previously
`
`determined) and fulvestrant because the estrogen receptor was being
`
`entirely bypassed.
`
`Id. at 0012 (“When taken together, these data provide
`
`InnoPharma Exhibit 1014.0010
`
`

`

`evidence
`
`for
`
`a mechanism by which FGF-stimulated estrogen
`
`independent grth bypasses the ER signal
`
`transduction pathway”).
`
`Because the estrogen receptor was being bypassed,
`
`it was not at all
`
`surprising that an estrogen receptor antagonist like fulvestrant would not
`
`inhibit tumor growth in that cell line.
`
`We observed in the McLeskey Reference that, despite the estrogen
`
`receptor bypass in the transfected breast cancer cell
`
`line, fulvestrant
`
`“retained activity” and worked exactly as intended in “preventing effects
`
`of endogenous estrogens” on the endometrium.
`
`Id. at 0006. And further,
`
`that it still worked in the cancer cells to inhibit the ER—however this ER
`
`inhibition was ineffective since the FGF pathway was bypassing the ER.
`
`Taken together, the McLeskey Reference would convey to the skilled
`
`researcher that fulvestrant effectively inhibits the estrogen receptor.
`
`Because of that known property, fulvestrant was used as a control in the
`
`McLeskey Reference to ensure that tumor growth could be attributed to
`
`fibroblast growth factor and not to estrogen, and to demonstrate that the
`
`ER expressed in the FGF-transfectants was still a functional ER. The
`
`fact that tumor growth was not inhibited in a transfected cell line where
`
`the estrogen receptor was being entirely bypassed would not in any way
`
`detract from those conclusions.
`
`InnoPharma Exhibit 1014.0011
`
`

`

`o
`
`I respectfully disagree with AstraZeneca and Dr. McLeskey”s assertion that
`
`the formulations identified in the McLeskey Reference would be interpreted
`
`as administrable to only animals. McLeskey Decl. 1l 6. Dr. McLeskey relies
`
`on tamoxifen pellets and a letrozole gavage in support of that assertion, but
`
`it
`
`is well-known that that tamoxifen and letrozole are solid oral dosage
`
`formulations. Mice cannot swallow pills, so solid oral doses must be given
`
`to mice as part of their food, which is inherently unreliable. As a result,
`
`scientists administer those drugs in a different form to animals, such as
`
`gavage or subcutaneous pellets,
`
`that results in reliable dosing.
`
`Those
`
`concerns do not apply to parenteral formulations, which can be administered
`
`to mice g humans. At the time,
`
`it was well-known that steroids were
`
`administered in oil depot
`
`formulations in humans, and fulvestrant
`
`is
`
`specifically described in the McLeskey Reference as “a preformulated drug”
`
`obtained from AstraZeneca.
`
`Id. at 0002. Given those facts, a skilled
`
`researcher would not interpret such a formulation as necessarily only an
`
`animal formulation.
`
`0
`
`I respectfully disagree with AstraZeneca and Dr. McLeskey’s conclusion
`
`that a skilled researcher “interested in developing a treatment for humans”
`
`would “not have looked at
`
`the McLeskey [Reference]
`
`for guidance.”
`
`McLeskey Decl.
`
`fll 11. Dr. McLeskey identifies three reasons for this
`
`InnoPharma Exhibit 1014.0012
`
`

`

`conclusion:
`
`(i)
`
`the McLeskey study was performed in mice;
`
`(ii)
`
`the
`
`McLeskey Reference explores hormone-independent breast cancer; and (iii)
`
`the McLeskey Reference allegedly provides no information about how to
`
`formulate fulvestrant.
`
`I briefly address these issues below.
`
`0
`
`I disagree that the use of mice in the McLeskey study somehow renders it
`
`irrelevant. Animal testing is a critical component of pre-clinical research,
`
`and researchers frequently rely on and use animal models to predict the
`
`efficacy and safety of drugs in humans.
`
`0
`
`In my view, a skilled researcher would assess hormone-independent and
`
`hormone-dependent pathways together—not separately as Dr. McLeskey
`
`asserts—to more effectively treat breast cancer. Specifically, fulvestrant
`
`was and is known to be a second-line therapy for the treatment of breast
`
`cancer, and is typically used after failure with tamoxifen.
`
`In order to
`
`effectively treat and understand breast cancer in a second-line setting, a
`
`skilled researcher would need to understand the mechanism for both
`
`hormone-dependent and hormone-independent breast cancer, and anti-
`
`estrogen sensitive and resistant breast cancer.
`
`Indeed,
`
`this directly
`
`impacts patient care. For example, a patient with hormone-independent
`
`cancer will likely be resistant to anti-estrogen therapy, and mechanisms
`
`leading to tamoxifen-resistance could cause cross-resistance to additional
`
`10
`
`InnoPharma Exhibit 1014.0013
`
`

`

`anti-estrogens like fulvestrant. Hormone-dependent cancer may become
`
`resistant during the course of first line treatment. A skilled researcher
`
`would need to understand both mechanisms to effectively treat such a
`
`patient.
`
`0 With respect to Dr. McLeskey’s criticism that the McLeskey Reference
`
`“provides no information about how to formulate fulvestrant,” l have
`
`been advised that the claims at issue here are directed to a method of
`
`treatment. Additionally, the McLeskey reference describes the specific
`
`formulation details for fulvestrant
`
`V.
`
`LEVEL OF ORDINARY SKILL IN THE PERTINENT ART
`
`17.
`
`I have been advised that
`
`there are multiple factors relevant
`
`to
`
`determining the level of ordinary skill in the pertinent art, including the educational
`
`level of active workers in the field at the time of the invention, the sophistication of
`
`the technology,
`
`the type of problems encountered in the art, and the prior art
`
`solutions to those problems.
`
`18.
`
`It is my opinion that a person having ordinary skill in the relevant art
`
`at the time of invention would have an advanced degree in medicine or a related
`
`field, with several years of practical experience in researching hormone dependent
`
`diseases of the breast.
`
`ll
`
`InnoPharma Exhibit 1014.0014
`
`

`

`VI. OVERVIEW OF TYPES OF BREAST CANCER AND THE
`
`AVAILABLE THERAPIES AS OF 2000
`
`19.
`
`Breast cancer is often described as either ER+ or ER — (and more
`
`recently HER2 assessed as well), with ER+ breast cancer then being hormone-
`
`dependent
`
`(i.e., hormone-sensitive) or hormone-independent
`
`(1'.e., hormone-
`
`insensitive).
`
`I discuss both of these forms of cancer below.
`
`A.
`
`Hormone-Dependent Breast Cancer
`
`20.
`
`In women, most invasive breast cancer cells are classified as ER+, and
`
`hormone-dependent, which means their growth is stimulated by hormones that
`
`occur naturally in women—primarily, estrogen and progesterone. These breast
`
`cancer cells contain receptors, or binding sites, that these hormones can attach to
`
`and activate, promoting cell growth and, ultimately, resulting in cancer.
`
`21.
`
`In the realm of hormone-dependent breast cancer, there are two main
`
`types of hormone receptors. They are estrogen receptors (ER) and progesterone
`
`receptors (PR).
`
`22.
`
`Various types of breast cancer are identified based on whether the
`
`cancer cells have hormone receptors. For example, hormone receptor-positive
`
`(HR+) breast cancer is generally hormone-dependent, while hormone receptor-
`
`negative (HR-) breast cancer is hormone-independent. More specifically, breast
`
`cancer cells with estrogen receptors are categorized as estrogen receptor-positive
`
`(ER+), and those without are called estrogen receptor-negative (ER-). Likewise,
`
`12
`
`InnoPharma Exhibit 1014.0015
`
`

`

`breast cancer cells with progesterone receptors are progesterone receptor-positive
`
`(PR+), and those without are progesterone receptor-negative (PR-).
`
`23.
`
`In order to treat HR+ breast cancer, doctors often prescribe hormone
`
`therapy, which functions by lowering the amount of hormones produced in the
`
`body or blocking hormones from attaching to receptors on breast cancer cells.
`
`This reduces or eliminates hormone receptor activation and, therefore, slows or
`
`stops cell growth.
`
`24.
`
`By 2000, the main types of hormone therapy were selective estrogen-
`
`receptor response modulators (SERMs), aromatase inhibitors (AIS), and selective
`
`estrogen-receptor downregulators (SERDS).
`
`I discuss each of these categories of
`
`hormone therapy below.
`
`(1)
`
`
`SERMs
`
`25.
`
`SERMs work by selectively binding to ERs in breast cells and cells in
`
`other estrogen target organs. In breast cancer cells, this results in antagonism such
`
`that the ER is thereby prevented from attaching to estrogen resulting in blocking of
`
`estrogen-induced cell growth.
`
`However, SERMs are not pure antagonists
`
`(inhibitors) and exhibit agonist (activator) activity depending on the identity of the
`
`cell.
`
`26.
`
`In other words, SERMs act as antagonists for ER in breast cells, but
`
`they can also activate estrogen receptors in other cells, such as bone, liver, and
`
`13
`
`InnoPharma Exhibit 1014.0016
`
`

`

`uterine cells. Ex. 1007 at 0001. Prior to 2000, the most commonly-prescribed
`
`SERM was tamoxifen.
`
`27.
`
`In addition to having antagonist activity, tamoxifen was known to
`
`have “agonist .
`
`.
`
`. activity as well.” Ex. 1039 at 0001. As a result, tamoxifen was
`
`linked to an increased risk of uterine cancer, and its use often resulted in tamoxifen
`
`resistance. Additional studies indicated that resistance to tamoxifen was not just a
`
`result of an alteration to the structure and function of the tamoxifen-estrogen
`
`receptor complex, but could also occur due to upregulated growth factor pathways
`
`that could either induce tamoxifen agonist activity or simply bypassed the ER.
`
`New pathways to cell growth that could affect ER and tamoxifen response
`
`included EGFR (epidermal growth factor receptor) signaling and FGFR (fibroblast
`
`growth factor receptor) signaling via fibroblast grth factors, discussed below.
`
`(2) fi
`
`28.
`
`In postmenopausal women, Als are used to stop the production of
`
`estrogen from the adrenal glands and fat tissue by blocking the enzyme aromatase.
`
`This enzyme converts the hormone androgen into estrogen in the body. The
`
`inhibition of estrogen production results in less estrogen to bind to estrogen
`
`receptors. However, AIs cannot block the production of estrogen in ovaries, so
`
`they are not effective in premenopausal women.
`
`29.
`
`Common Als include Arimidex® and Femara®. Although AIs do not
`
`14
`
`InnoPharma Exhibit 1014.0017
`
`

`

`have the same agonist properties as tamoxifen,
`
`they are associated with an
`
`increased risk of bone loss.
`
`(3)
`
`
`SERDs
`
`30.
`
`SERDs function by binding to estrogen receptors and preventing
`
`estrogen from attaching to and activating the estrogen receptor.
`
`In addition,
`
`SERDs break down or change the shape of estrogen receptors in breast cells
`
`resulting in its degradation, such that
`
`there are fewer
`
`functioning estrogen
`
`receptors to interact with estrogen.
`
`31.
`
`ICl 182,780, or fulvestrant, was developed in the 1990s as an
`
`alternative to tamoxifen. Ex. 1031 at 0001. It was well-known that fulvestrant—a
`
`steroidal SERD—has significant antiestrogenic potency and functions as a pure
`
`antagonist that blocks the activation of estrogen receptors.
`
`Ids, Ex. 1039 at 0001',
`
`Ex. 1007 at 0001. Because fulvestrant is a pure antagonist with no agonist activity,
`
`it was known as an effective, if not superior, alternative to tamoxifen, especially as
`
`second-line therapy following tamoxifen resistance. Ex. 1007 at 0007.
`
`B.
`
`Hormone-Independent Breast Cancer
`
`32. While many patients respond to hormone therapy,
`
`there are also
`
`patients who either fail to show any initial response to hormone-based treatment or
`
`later acquire resistance to such therapy. Ex. 1069 at 0003. Researchers classify
`
`breast tumors that do not depend on hormones for growth as hormone-independent
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`InnoPharma Exhibit 1014.0018
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`breast cancer.
`
`33. Hormone independent breast cancer may result from growth factor
`
`signaling. Growth factors are naturally occurring substances that can stimulate cell
`
`growth. Two main types of growth factors that are usually associated with breast
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`cancer are epidermal growth factor receptor (EGFR), human epidermal growth
`
`factor receptor 2 (HERZ), and fibroblast growth factor (FGFR).
`
`34. Human epidermal growth factor receptor 2 (HERZ) is a protein found
`
`on breast cells that controls how cells grow, divide, and repair themselves.
`
`Overexpression of HER2 can cause breast cells to divide uncontrollably, leading to
`
`breast cancer.
`
`35.
`
`Like HERZ, fibroblast growth factors (FGFS) are essential proteins
`
`found in healthy cells, but overexpression of these growth factors or their receptor
`
`can lead to tumor growth. FGFs stimulate the growth of blood vessels in a tumor,
`
`providing oxygen and nutrients to promote cell growth and helping spread the
`
`tumor throughout the body, as well as promote growth pathways within cells. Ex.
`
`1069 at 0007.
`
`36.
`
`By 2000, a role for growth factor signaling in tamoxifen resistance
`
`was known.
`
`In fact, expression of growth factors, including FGF, provided an
`
`alternative pathway for tumor growth by entirely “bypass[ing] the need for ER
`
`activation” in ER+ cancer cells. Id. at 0008', Ex. 1008 at 0002.
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`37.
`
`Before the McLeskey Reference, researchers hypothesized that FGF-
`
`mediated breast cancer could result in a change to the conformation of the estrogen
`
`receptor, or that it might bypass the estrogen receptor entirely. Since fulvestrant
`
`was used as a second-line therapy for
`
`tamoxifen resistance, we needed to
`
`investigate whether FGF-mediated cell
`
`lines would be
`
`cross-resistant1 to
`
`fulvestrant and aromatase inhibitors. We set out to test this hypothesis in the
`
`McLeskey Reference.
`
`VII. OVERVIEW OF THE MCLESKEY REFERENCE
`
`A.
`
`Summagy of the Testing Conducted in the McLeskey Reference
`
`38. We set out to test that hypothesis in the McLeskey Reference.
`
`In
`
`particular, we wanted to understand whether
`
`fibroblast growth factor was
`
`“replacing estrogen as .
`
`.
`
`. stimulus for tumor growth.” Id. at 0001. To confirm
`
`that conclusion, we utilized MCF—7 cell lines.
`
`Id. at 0002. The MCF-7 breast
`
`cancer cell line—named after the Michigan Cancer Foundation—is a human breast
`
`cancer cell line that was obtained from a postmenopausal woman in the 1970s.
`
`It
`
`is often used to study the estrogen receptor because of its sensitivity to estrogen
`
`manipulation.
`
`1 Cross-resistance refers to being resistant
`
`to both tamoxifen and additional
`
`antiestrogens such as fulvestrant.
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`39.
`
`For purposes of the study, we introduced a modification into the
`
`MCF-7 cell line.
`
`In particular, we modified this cell line to overexpress FGF so
`
`that we could carefully study FGF and its
`
`impact on cellular
`
`signaling.
`
`Throughout the study, we called our modified cell line the “FGF-transfected MCF-
`
`7 cell line.”
`
`40.
`
`During the study, we injected the FGF-transfected MCF-7 cell line
`
`into ovariectomized mice (i.e., mice with their ovaries
`
`removed).
`
`It
`
`is
`
`commonplace for researchers to rely on animal models—and mouse models in
`
`particular—for
`
`studying human breast cancer.
`
`Because of “advances
`
`in
`
`manipulating targeted genes in a tissue-specific manner,” mouse models can play a
`
`valuable role in studying the molecular pathways involved in human breast cancer
`
`progression without directly experimenting on human patients. EX. 1065 at 0001.
`
`Even though a mouse model was used in our study, the research was directed
`
`toward elucidating aspects of human breast cancer.
`
`41.
`
`To confirm the hypothesis that FGF—rather than estrogen—was
`
`stimulating tumor growth in FGF-mediated cell lines, we needed to “abrogate all
`
`estrogenic activity” in the mice. Ex. 1008 at 0010. Although the mice had their
`
`ovaries removed, estrogen synthesized “at extraovarian sites” was still expected to
`
`be present.
`
`Id. at 0004. Thus, we need to eliminate this extraovarian estrogen to
`
`ensure that it was not playing a role in stimulating tumor growth. Id.
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`42.
`
`To do so, we administered the most efficacious antiestrogens known
`
`at the time—fulvestrant and two aromatase inhibitors—in separate experiments in
`
`the McLeskey Reference. 161., see also McLeskey Decl. 11 5 (fulvestrant was used
`
`to “make sure that the estrogen receptor (ER) was not activated by small amounts
`
`of estrogen synthesized by the mouse’s liver and adrenal glands—with the goal
`
`being to determine if the activity of FGF (rather than estrogen) could drive tumor
`
`growth in tamoxifen-resistant breast cancer cells”). We knew that fulvestrant
`
`would be effective in inhibiting all estrogenic activity because we had previously
`
`confirmed fulvestrant’s activity in tests on a control cell line, which we refer to in
`
`the McLeskey Reference as the ML—20 cell line. Id. at Figure 4, 5, n. 4.
`
`43.
`
`In the study, we used two different formulations of fulvestrant: (l)
`
`“powdered [fulvestrant] dissolved in 100% ethanol and spiked into warmed peanut
`
`oil” to give a final concentration of 50 mg/mL” and (2) “50 mg/mL drug in a
`
`vehicle of 10% ethanol, 15% benzyl benzoate, 10% benzyl alcohol, brought to
`
`volume with castor oil.” EX. 1008 at 0002. The latter formulation was supplied to
`
`us “preformulated” directly from AstraZeneca. Id.
`
`B.
`
`Results Reported in the McLeskey Reference
`
`44.
`
`For at least four reasons, the results that we reported in the McLeskey
`
`Reference confirm that
`
`fulvestrant worked exactly as
`
`it was
`
`intended—by
`
`inhibiting estrogenic activity.
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`45.
`
`First, we directly confirmed that fulvestrant was blocking the estrogen
`
`receptors in the FGF cells.
`
`In particular, as shown in Figure 8 in the McLeskey
`
`Reference,
`
`treatment of FGF-expressing cells with fulvestrant
`
`in estrogenic
`
`conditions abrogates estrogen-induced elevated ER activity.
`
`Id. at 0011. These
`
`results confirm that ER is still activated when estrogen is available (i.e., in the
`
`absence of fulvestrant), and that fulvestrant successfully blocks estrogen from
`
`triggering ER activation.
`
`46.
`
`The results also confirm that FGF transfection does not directly
`
`stimulate any ER activity. Accordingly, we concluded that “the transfected FGFs
`
`are stimulating growth by a mechanism that bypasses the ER-mediated growth-
`
`stimulatory pathway.”
`
`Id. at 0010.
`
`In other words, because we were able to
`
`confirm that fulvestrant was successfully blocking the ERs in the FGF cells, we
`
`knew that the FGF-mediated cancer cell growth was mediated by a pathway
`
`independent of the ER. The diagram below illustrates this point:
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`ER-D epen dent
`Pathway
`
`Response: C ell Growth
`
`Pathway .
`
`ER Bypass
`Pathway
`
`I FGF Receptor
`
`ER-Depen dent
`
`r’”“"~\
`fix
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`\
`
`47.
`
`Second, we confirmed that
`
`fulvestrant “retained activity” and
`
`inhibited endometrial growth in intact mice, which demonstrates that fulvestrant
`
`was blocking the estrogen receptors in these mice.
`
`Specifically, we injected
`
`fulvestrant into reproductively intact female mice for two weeks “to observe for
`
`activity in preventing effects of endogenous estrogens on the endometrium,” or
`
`uterine lining. Id at 0005-06. Results from this experiment showed that the mice
`
`treated with fulvestIant
`
`lacked endometrial glandular structures.
`
`Id.
`
`Thus,
`
`fulvestrant “retained activity” and worked exactly as intended in preventing growth
`
`of endometrial cells, even though it had no effect on breast tumor growth in
`
`experiments with FGF-transfected breast cancer cells. Id.
`
`48.
`
`Third, we confirmed with control DAL-20 cells
`
`that no growth
`
`occurred in the presence of fulvestrant. Ex. 1008 at Figure 4, Figure 5.
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