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`PHIGENIX
`PHIGENIX
`Exhibit 1029
`Exhibit 1029
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`

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`THE UNITED STATES PATENT AND TRADEMARK OFFICE
`
`Applicant:
`
`Sharon Erickson
`
`Attorney Docket #:
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`GNE-0073R2C2
`
`Serial No.
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`11/949,351
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`Group Art Unit
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`1643
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`Filing Date
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`12/03/2007
`
`Examiner:
`
`Nataraj an, Meera
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`Customer No.:
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`35489
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`Confirmation No.:
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`4598
`
`Title:
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`METHODS OF TREATMENT USING ANTI-ErbB ANTIBODY
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`MAYTANSINOID CONJUGATES
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`FILED VIA EFS
`
`Commissioner for Patents
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`PO. Box 1450
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`Alexandria, VA 22313—1450
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`DECLARATION OF BARBARA KLENCKEa MD.
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`1, Barbara Klencke, M.D., declare and say as follows: -
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`l.
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`I obtained a BS. in 1979 from Indiana University, and an MD. from University
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`of California, Davis. I completed my residency at University of California San Francisco
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`(UCSF) Medical Center. Following two years of Medical Oncology fellowship training at
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`UCSF, I served as Assistant Professor of Medicine for over seven years at UCSF.
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`2.
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`I joined Chiron (later acquired by Novartis) in 2002 and served as Medical
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`Director for one year, and in 2003, Ijoined Genentech, Inc., where I worked as Associate Group
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`Director, and where my current title is Group Director.
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`3.
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`During my employment at Genentech, I have worked on the development of
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`marketed products, such as Avastin® and Tarceva®, and several therapeutic compounds in early
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`development, prior to working on the development of T—DMl. T-DM1 is a therapeutic
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`immunoconjugate in which the anti—ErbB2 antibody trastuzumab is conjugated to the cytotoxic
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`maytansinoid “DMl.” Part of my responsibilities as Associate Group Director included the
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`PHIGENIX
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`Exhibit 1029-01
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`

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`development of clinical trial strategy for T-DMl in the treatment of tumors, primarily breast
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`tumors, that express ErbB2.
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`4.
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`My Curriculum Vitae, including my list of publications, patents, pending patent
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`applications, and awards, is enclosed as Exhibit A and forms part of this Declaration.
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`5.
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`I am familiar with and understand the disclosure of the above—identified patent
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`application, including the currently pending claims, including the new claims added concurrently
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`with filing the present Declaration. I am also familiar with and understand the Office Action
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`mailed on June 8, 2010 in connection with the above—identified patent application, and the
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`references cited in that Office Action. Claim 40 is directed to an immunoconjugate comprising
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`the antibody huMAb4D5—8 conjugated to a maytansinoid. The antibody huMAb4D5—8 is the
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`same antibody as trastuzumab and is marketed under the tradename Herceptin®. All other claims
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`depend, directly or indirectly, from claim 40.
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`6.
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`According to the Office Action, the claimed invention would have been obvious
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`to one of ordinary skill in the art at the time the invention was made over the combination of
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`U.S. Patent No. 5,208,020 (referred to hereinafter as “Chari et al.”) and U.S. Patent No.
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`6,054,297 (referred to hereinafter as “Carter et al.”). Chari et a1. allegedly teach a composition
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`comprising one or more maytansinoids (col. 6-8) linked to a monoclonal antibody or antibody
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`fragment, where the monoclonal antibody is selective for tumor cell antigens. Carter is cited for
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`disclosing humanized 4D5 antibodies, including huMAb4D5—8, and for allegedly teaching that
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`the humanized 4D5 antibodies may be used as immunotoxins, conjugated with a cytotoxic
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`moiety (col. 44). The Examiner acknowledges that Chari et a1. do not teach an anti—ErbB2
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`antibody conjugated to a maytansinoid. The finding of obviousness is based on the assertion that
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`one of ordinary skill would have been motivated to make the claimed antibody—maytansinoid
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`conjugates because such conjugates fall within the scope of the immunotoxins of Carter et al. and
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`because Carter et al. teach that ErbB2 is amplified or overexpressed in human malignancies.
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`7.
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`Before the earliest priority date of the present application (March 16, 2000), it
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`was known that ErbB2 (also known as HER2) is amplified and overexpressed in certain breast
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`and ovarian cancers, and such overexpression is correlated with poor prognosis. (See application
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`at page 2, para. [0006].) It was also known that HER2 is expressed in normal tissues at levels
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`PHIGENIX
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`Exhibit 1029-02
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`similar to those found in non-HER2-amplified, non—HERZ—overexpressing breast cancers and
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`breast cancer cell lines. For example, HER—2 protein was identified on cell membranes of
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`normal epithelial cells in the gastro—intestinal, respiratory, reproductive, and urinary tract as well
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`as in the skin, breast and placenta, demonstrating that HER2 is normally a membrane constituent
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`of a variety of epithelial cell types.
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`(See, Press et al., Oncogene 5(7):953—62, 1990 — Exhibit B,
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`Abstract, Table 1 and Figures 1 and 2.)
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`8.
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`Herceptin® was approved by the FDA in 1998 for the treatment of metastatic
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`HERZ—overexpressing breast cancer, either as an initial treatment in combination with
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`chemotherapy (paclitaxel), or as a monotherapy after prior treatment with chemotherapy.
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`Herceptin® was later approved in 2006 as part of a multi—agent regimen for the adjuvant (post—
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`surgical) treatment of HERZ—overexpressing breast cancer. In 2008, Herceptin® received further
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`approvals from the FDA for treatment in the adjuvant setting. Although Herceptin® is a
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`breakthrough in the treatment of HERZ—overexpressing (“HER2-positive”) breast cancer, nearly
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`all patients with metastatic disease treated with Herceptin® will progress after experiencing a
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`period of clinical benefit, and there remains an unmet medical need for patients with HER2—
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`positive breast cancer. (See application at page 5, para. [0014].)
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`9.
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`The maytansinoids are a family of cytotoxic molecules that include maytansine
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`and its derivative DM1. Before the priority date, it was reported that maytansine acts as a very
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`potent mitotic inhibitor by inhibiting microtubule polymerization. (See abstract, Rao et al.,
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`Cancer Research 39:3152—3155, 1979 — Exhibit C; and see abstract, Remillard et al., Science
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`189: 1002—1005, 1975 — Exhibit D.) Maytansine was also reported as being of relatively high
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`cytotoxicity (i.e., about 100—200 fold more cytotoxic than other anti—mitotic agents). (See
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`Exhibit C, col. 1, para. 2.) Maytansine was unsuccessful in human clinical trials because of
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`unacceptable systemic toxicity, i.e., maytansine did not demonstrate clinically reasonable
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`therapeutic benefit at safe doses. (See Issell et al., Cancer Treat. Rev. 5:199—207, 1978 — Exhibit
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`E, e.g., at pages 204 and 207.)
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`10.
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`Before the earliest priority date of the present application, it was recognized in the
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`art that some monoclonal antibody-based therapies had begun to show promise in the treatment
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`of cancer, as demonstrated, e.g., by Herceptin® and by Rituxan®, another Genentech monoclonal
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`PHIGENIX
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`Exhibit 1029-03
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`antibody first approved in 1997 for the treatment of non—Hodgkin’s lymphoma. Despite the
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`promise shown by some monoclonal antibodies in the clinical setting at the priority date,
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`immunoconjugates (referring to antibodies conjugated to moieties such as radioisotopes,
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`enzymes, drugs, and cytotoxic agents) had achieved very limited clinical success at that time, a
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`fact that holds‘true to this day. Indeed, Trail and Bianchi recognized at the priority date that
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`immunoconjugates were not “established” chemotherapeutic agents and that further work in the
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`field was needed to improve “the efficacy and therapeutic index of immunoconjugates by
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`optimizing selectivity and potency.” (See Trail and Bianchi, Current Opinion in Immunology
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`11:584—588, Oct. 1999 — Exhibit F, e.g., at page 586, col. 2, para. 2.) “Selectivity” refers to the
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`ability of immunoconjugates to bind preferably to tumor cells expressing the target antigen over
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`normal cells that may also express the target antigen, while “potency” refers to the degree of
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`cytotoxicity of the immunoconjugate, as conferred by the cytotoxic moiety of the
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`immunoconjugate. Trail and Bianchi cautioned that “[i]t is therefore necessary to balance the
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`relative selectivity of the MAb [monoclonal antibody] with the potency of the agent delivered.”
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`(See Exhibit F page 584, col. 1, para. 2.) If that balance is not achieved, an immunoconjugate
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`may cause toxicity on the one hand, or may not be sufficiently potent on the other.
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`(See Exhibit
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`F, page 585, col. 1, first para, and col. 2, last para.) Indeed, Trail and Bianchi further cautioned
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`that “even low levels of expression of the targeted antigen by normal cells may lead to significant
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`toxicity.” (Exhibit F, page 585, col. 1, para. 1, emphasis added.)
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`11.
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`Thus, at the priority date, the immunoconjugate field was still evolving with no
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`outstanding successes, and there was considerable uncertainty and concern surrounding the
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`therapeutic potential and in particular, the therapeutic index, of immunoconjugates. Indeed, Trail
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`and Bianchi predicted that the major role of immunoconjugates would lie in “minimal disease
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`settings,” based on the assumption that potency would need to be comprised in order to avoid
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`unacceptable levels of toxicity. (See Exhibit F, e. g., at page 585, C01. 2, last para, and page 586,
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`col. 2, last para.)
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`12.
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`In my capacity as Associate Group Director at Genentech, I played a major role in
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`designing the “TDM4374g” clinical trial protocol. The TDM4374g clinical trial is a
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`Genentech/Roche—sponsored single-arm, multi—center Phase II trial of T—DMl in 110 patients
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`with HER2—positive metastatic breast cancer. Patients that participated in this study had
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`PHIGENIX
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`Exhibit 1029-04
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`progressive disease after having been treated with at least the following five different agents or
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`classes of agents: an anthracycline, a taxane, capecitabine (Xeloda®) and two HERZ-directed
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`therapies, i.e., Herceptin® and lapatinib (Tykerb®, an inhibitor of the intracellular tyrosine
`kinase domain of HERZ). Anthracyclines, taxanes and capecitabine encompass the most
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`commonly used chemotherapeutic agents in the treatment of breast cancer; Herceptin® and
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`lapatinib represent the only FDA-approved HERZ—direeted therapies for metastatic HERZ-
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`positive breast cancer. Thus, the patients in the TDM4374g trial had been previously treated
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`with at least five agents representing the most commonly used classes of drugs currently
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`available for the treatment of metastatic breast cancer, including the only two that are approved ‘
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`for the treatment of HERZ—positive breast cancer.
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`13.
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`The patients enrolled in the TDM4374g trial had been treated with a median of
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`seven different agents, yet their cancers had progressed after the last therapy received. T—DMl
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`was essentially a last option for the terminally ill patients in this study.
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`14.
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`At this point in the TDM4374g trial, the clinical activity of T—DMl has well
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`exceeded what a clinical oncologist practicing in the field of metastatic breast cancer would have
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`expected, given the clinical setting the trial was designed to address as well as the activity of
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`other agents currently used in the treatment of metastatic breast cancer.
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`15.
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`In the TDM4374g trial as of December 2009, the objective response rate (ORR),
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`which is the percentage of patients whose tumors shrank by at least 30% after treatment with T—
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`DMl, was 32.7%, as determined by an independent review facility. The clinical benefit rate,
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`which is the percentage of patients with objective response or stable disease maintained for at
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`least 6 months, was 44.5%, as determined by an independent review facility.
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`16.
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`To put these data in perspective, patients initially presenting with HERZ—positive
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`metastatic breast cancer typically receive Herceptin® in combination with a taxane. If a patient’s
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`disease progresses after that “first—line” therapy, then a “second-line” HERZ—directed therapy is
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`typically administered. That second—line therapy typically consists of either Herceptin® or
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`lapatinib administered in combination with further chemotherapeutic agents. After first— and
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`second—line therapies fail, the treatment options for metastatic breast cancer are very limited.
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`PHIGENIX
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`Exhibit 1029-05
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`One remaining approved treatment option in this “third—line” setting is ixabepilone (Ixempra®;
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`Bristol—Myers Squibb), a non—HER2—specific microtubule inhibitor.
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`17.
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`The ORR for T—DMl was unexpectedly better than the ORR seen with agents
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`currently used in second— and third-line metastatic breast cancer therapies. For example, the
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`ORR for the combination of lapatinib and capecitabine was 23.7% in a “second-line” study of
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`patients that had received prior treatment with an anthracycline, a taxane and Herceptin®.
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`(See
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`Tykerb® package insert, rev. 2010 — Exhibit G, e. g., at page 15 , Table 5.) The ORR for
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`ixabepilone in the treatment of metastatic breast cancer previously treated with an anthracycline,
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`a taxane and capecitabine was 12.4%. (See Ixempra® package insert, rev. 2009 — Exhibit H,
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`e.g., at Table 8.)
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`18.
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`Based on my experience as an oncologist, the trial would have been considered a
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`success if T—DMl, which was administered as a third—line therapy in TDM4374g, demonstrated
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`an ORR comparable to that of current second line therapies. Thus, an ORR of about 24% would
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`have been considered a favorable outcome in the TDM4374g trial. The fact that the ORR of T—
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`DMl in TDM4374g (32.7%) significantly exceeded that of current second—line therapies
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`(23.7%), and well surpassed that of current third line therapies (12.4%), was a better result than
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`expected.
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`19.
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`T—DMl also showed considerably low systemic toxicity. T—DMl was generally
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`well tolerated by patients at the dose and schedule tested, and the observed toxicities were
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`generally acceptable and manageable in this patient population. The low toxicity of T—DMl is an
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`advantage, particularly in light of the fact that HER2 is expressed on normal cells, and that
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`maytansine (from which DMl is derived) had demonstrated unacceptable toxicity in previous
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`clinical trials.
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`20.
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`It is my considered opinion that T—DMl provides an important advance in the
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`field of HER2—positive metastatic breast cancer treatment, by showing unexpectedly superior
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`activity in a patient population that had progressed after treatment with multiple standard
`chemotherapies as well as all currently approved HER2—directed therapies. TDM4374g is the
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`first clinical trial that specifically treated patients who had progressive disease after receiving
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`five FDA—approved agents or classes of agents for the treatment of breast cancer, including the
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`PHIGENIX
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`Exhibit 1029-06
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`only two approved HER2—directed agents (trastuzumab and lapatinib). That patient population
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`has very limited treatment options. Current chemotherapeutic options in this setting have poor
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`response rates and significant toxicity. In contrast, T—DMl provides a Viable single treatment
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`option with impressive efficacy and a favorable safety profile.
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`21.
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`The opinion that T-DMl demonstrated unexpectedly superior activity is reflected
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`by the statements of others in the field praising the performance of T-DMl in the TDM4374g
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`trial. For example, Dr. Ian Krop, the lead clinical investigator in the study, was quoted in a
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`“WebMD” online article as stating that “[t]his is the first study looking at women who have
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`failed so many other treatments” and “we think these results are as good as we’ve ever seen in
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`such a refractory population.” (See. WebMD website http://Www.webmd.com/breast—
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`cancer/news/20091215/targeted—breast-cancer—drug—shrinks—tumors, dated 12/ 1 5/2009, printed on
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`12/ 18/2009 — Exhibit I, emphasis added.) Dr. Krop was also quoted in an online article in
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`“TheStreet.com” as stating: “T-DMl is a drug that I’d like to see made available to patients. I’m
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`sure Genentech [Roche] is strongly considering using this data for the basis of an accelerated
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`approval filing.” (See http://www.thestreet.com/print/story/ 10644637.html, dated 12/14/2009,
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`printed on 12/ 18/2009 — Exhibit J, emphasis added.)
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`22.
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`Edith Perez, M.D., a breast cancer specialist and internationally known researcher
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`at the Mayo Clinic, was quoted in the “WebMD” online article cited above as stating that “[t]he
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`response rate they saw in the study is exceptional in a group of patients this ill.” (See Exhibit 1,
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`emphasis added.) (It is noted that Dr. Perez was not a clinical investigator in the TDM4374g
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`trial but is an investigator in other Genentech/Roche—sponsored clinical trials of T—DMl in other
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`settings.)
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`23.
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`It is also my considered opinion that the results of the TDM4374g trial challenged
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`the skepticism surrounding immunoconjugates at the priority date, e.g., that the major role of
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`immunoconjugates would lie “in minimal disease settings.” In the TDM4374g trial, T—DMI
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`demonstrated unexpectedly superior efficacy as a single agent in an extremely advanced disease
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`setting, thus well exceeding expectations at the priority date. T—DMl achieved this feat by
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`demonstrating both impressively high potency and low toxicity. Those results were not
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`PHIGENIX
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`Exhibit 1029-07
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`

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`predictable, given that maytansinoids are highly cytotoxic molecules and that HER2 is expressed
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`on normal cells.
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`24.
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`It is further my considered opinion that T—DMl fills a long felt but unresolved
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`need at the priority date, i.e., the need for HER2—directed agents that treat metastatic HERZ—
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`positive breast cancer. At the priority date, only one HERZ—directed therapy, i.e., Herceptin®,
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`had been approved by the FDA for the treatment of HERZ—overexpressing metastatic breast
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`cancer. Moreover, nearly all patients with metastatic disease treated with Herceptin® will
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`progress after experiencing a period of clinical benefit.
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`25.
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`The opinions expressed in paragraphs 20 and 23—24 are consistent with those of
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`other experts in the field, who have praised T—DMl and would like to see T—DMl available to
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`patients (i.e., approved by the FDA) as soon as possible. As shown by the results of the
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`TDM4374g trial, T—DMl has the potential to considerably improve the lives of breast cancer
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`patients, offering new hope to patients suffering with advanced, aggressive disease.
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`26.
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`It is also my considered opinion that T—DMl is a pioneer in the immunoconjugate
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`field. To this day (about 10 years after the priority date), immunoconjugates have achieved very
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`limited clinical success. Although immunoconjugates have been subject to over two decades of
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`research and development within the scientific and pharmaceutical community, the FDA has thus
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`far approved only one immunoconjugate, Mylotarg®, for the treatment of acute myeloid
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`leukemia, a hematologic cancer. The unexpectedly superior activity of T—DM 1 in treating solid
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`tumors represents an important advance in the field of immunoconjugates.
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`27.
`I declare further that all statements made in this Declaration of my own
`knowledge are true and that all statements made on information and belief are believed to be true
`and further, that these statements are made with theknowledge that willful statements and the
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`like so made are punishable by fine or imprisonment, or both, under Section 1001 of Title 18 of
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`the United States Code and that such willful false statements may jeopardize the validity of the
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`application or any patent granted thereon.
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`
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`PHIGENIX
`
`Exhibit 1029-08
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`

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`EXHIBIT A
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`PHIGENIX
`
`xhibit 1029-09
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`

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`Barbara J. Klencke, M.D.
`
`Genentech, Inc.
`1 DNA Way, M/S 66
`South San Francisco, CA 94080
`(650) 467—6278 Phone
`e-mail: bklencke@gene.com
`
`Emeloyment
`
`2010 - present
`2003 - 2010
`
`Group Medical Director
`Associate Group Medical Director
`BioOncology
`Genentech, Inc.
`South San Francisco, CA
`
`Details of Genentech Career History
`2003 - 2005: Assistant then Associate Medical Director, Tarceva team, GNE
`2005:
`Associate Medical Director, Exploratory Clinical Development, GNE
`2005 - 2007: Medical Director then Senior Medical Director then Associate Group Medical
`Director; Avastin Breast Team
`2007 - 2008: Associate Group Medical Director: T—DM1 team and Development Subteam Lead
`for T—DM1
`
`2002 —— 2003
`
`Medical Director, Clinical Development
`Chiron Corporation
`Emeryville, California
`
`1998 — 2002
`
`Assistant Clinical Professor of Medicine
`
`University of California at San Francisco
`Division of Hematology/Oncology, Department of Medicine
`
`1995 - 1998
`
`Clinical Instructor
`
`University of California at San Francisco
`Division of Hematology/Oncology, Department of Medicine
`
`Education
`
`
`
`1993 - 1995
`
`University of California at San Francisco
`Medical Oncology Fellowship
`
`PHIGENIX
`
`Exhibit 1029-10
`
`

`

`1990 - 1993
`
`University of California at San Francisco
`Internship and Residency
`Internal Medicine, Categorical Program
`
`1986 - 1990
`
`University of California at Davis School of Medicine
`Degree: MD.
`
`1984 — 1985
`
`University of California at Davis
`
`1975 — 1979
`
`Indiana University at Bloomington
`Degree: BS
`
`Honors and Awards
`“
`
`1989
`1986 ~ 1990
`1990
`
`1995
`
`Alpha Omega Alpha
`Co-Class President, University of California Davis, School of Medicine
`Graduation Awards, University of California Davis, School of Medicine
`- Department of Medicine Award
`- American Medical Women’s Association Award
`
`- Lange Publication Award
`American Federation of Clinical Research
`Excellence in Clinical Research Award
`
`Board Certifications, Licensure, Prolessional Sociez Memberships
`
`1993
`
`1995
`
`G74133
`
`Member:
`
`Internal Medicine
`
`Medical Oncology
`
`_
`
`California State Medical License; issued May 26, 1992
`
`ASCO since 1995
`AACR since 2001
`ASH since 2002
`
`Committee eL‘Zorts, Teaching, Public Service
`
`
`2002 — 2002
`
`Chair, HPV Working Group
`AIDS Malignancies Clinical Trials Consortium (AMC)
`
`1999 — 2001
`
`C0— Chair, HPV Working Group, AMC
`
`1999 — 2002
`
`Steering Committee, AMC
`
`1998 — 2002
`
`Kaposi’s Sarcoma Working Group, Member, AMC
`
`1997 — 2002
`
`Director, Oncology Educational Electives for Residents, UCSF
`
`1999 — 2002
`
`UCSF Hematology / Oncology Fellowship Evaluation Committee, Member
`
`2
`
`PHIGENIX
`
`Exhibit 1029-11
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`

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`1999- 2002
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`Protocol Review Committee, UCSF Cancer Center, Member
`
`1997 — 1998
`
`Dean’s Advisory Committee for AIDS, UCSF, Member
`
`1999 — 2002
`
`Advisory Committee to the UCSF AIDS Research Institute, Member
`
`1999 - 2000
`
`Patient / Physician Communication Task Force
`National Comprehensive Cancer Network
`
`V
`Wblico .
`
`
`
`Manuscripts
`
`l.
`
`10.
`
`11.
`
`W. K. Alfred Yung, James J. Vredenburgh, Timothy F. Cloughcsy, Phioanh (Leia) Nghiemphu,
`Barbara Klencke, Mark R. Gilbert, David A. Reardon, Michael D. Prados. Safety and Efficacy of
`Erlotinib in First—Relapse Glioblastoma: A Phase II_Open—Label Study. Submitted (Oct 2008) for
`publication to NeuroOncology.
`Chan AT. Tao Q. Robertson KD. Flinn IW. Mann RB. Klencke B. Kwan WH. Leung TW. Johnson
`PJ. Ambinder RF. Azacitidine induces demethylation of the Epstein—Barr virus genome in tumors.
`Journal of Clinical Oncology. 22(8):1373-81, 2004 Apr 15
`Klencke B, Palefsky J. Anal Cancer: An HIV-related malignancy. Hematology and Oncology
`Clinics of North America. 17(3):859-72, 2003 Jun.
`Rose B, Matthay KK, Price D, Huberty J, Klencke B, Norton JA, Fitzgerald PA. High dose 131I-
`Metaiodobenzylguanadine (MIGB) Therapy for 12 Patients with Malignant Pheochromocytoma.
`Cancer 2003; 98(2): 23 9—248.
`_
`Rodrigues L, Klencke B, Vin-Christian K, Berger TG, Crawford RI, Miller JR 3rd, Ferreira CMM,
`Nosrati M, and Kashani-Sabet M. Altered Clinical Course of Malignant Melanoma in HIV-Positive
`Patients. Archives of Dermatology 2002; 138:765—770.
`_
`Klencke B, Urban RG, Berry M, Matijevic, M, Lathey JL Thatcher J, Wilson J, Jay N, Palefsky J.
`Microsphere Encapsulated Plasmid DNA (pDNA) Treatment for HPV-16 Associated Anal
`Dysplasia: A Phase I Study of ZYClOl. Clin Cancer Res 2002 8: 1028-1037.
`Benson HI AB, Mitchell E, Abramson N, Klencke B, Ritch P, Burnham JP, McGuirt C, Bonny T,
`Levin J, Hohneker J. Oral eniluracil/fluorouracil in patients with inoperable hepatocellular
`carcinoma. Ann Oncol. 2002 Apr;13(4):576-81.
`Blackbourn DJ, Lennette E, Klencke B, Moses A, Chandran B, Weinstein M, Glogau RG, Witte
`MH, Way DL, Kutzkey, Herndier D, and Levy JA. The Restricted Cellular Host Range Of HHV—8.
`AIDS 2000; 14(9):1123-1133.
`Hoffman R, Krieg R, Klencke B, Weinberg V, Welton ML. The significance of pretreatment CD4
`count on the outcome and treatment tolerance of HIV-positive patients with anal cancer. Int. J. of
`Radiation Oncology Biol., Phys. 1999; 44(1): 127—3 1.
`Klencke B, Kaplan L. Advances and future challenges in Hon-Hodgkin's lymphoma. Current
`Opinion in Oncology 1998; 10: 422—427.
`Dorsey BV, Benjamin LE, Rausher F, Klencke B, Venook AP, Warren RS, Weidner N. Intra-
`Abdominal Desmoplastic Small Round—Cell Tumor: Expansion of the Pathologic Profile. Modern
`Pathology 1996; 9 (6):703—709.
`
`PHIGENIX
`
`Exhibit 1029-12
`
`

`

`Abstracts
`
`Brooks DJ, Cox J, Berman BS, Amare M, Benson AB, Masuoka L, Klencke B. Phase 2 study of
`tezacitabine in subjects with advanced/recurrent colorectal cancer. PASCO 2003, abstract 1122.
`Klencke B. HIV-Associated Anal Cancer: Biology, Epidemiology, Prevention and Treatment.
`PASCO, Educational Session, May 2002.
`Klencke B, Urban RG, Berry M, Thatcher J, Wilson J, Matijevic, M, Lathey JL, Jay N, Palefsky J.
`Results from a Phase I Trial of a Microsphere Encapsulated DNA Vaccine (ZYC101) for High-grade
`Anal Dysplasia. Oral Presentation. Fifth International AIDS Malignancies Conference, April 23-25,
`2001.
`
`Klencke B. HPV in Anal and Penile Cancers. Papilloma virus: 18th International Conference.
`Barcelona July 21-28, 2000. Oral Presentation
`Klencke B, Berry JM, Thatcher J, Darraugh T, Rokovich J, Urban R, Palefsky J. Phase I Clinical
`Trial of HPV-1 6 E7 Vaccine for High Grade Anal Dysplasia. WHO / Eurogin Human
`Papillomavirus, Genital infections and Neoplasia Conference. Paris April 5—9, 2000. Oral
`Presentation 4/9/2000.
`
`Martin, J., Lee, T., Busch, M., Cheng, A., Graves, A., Klencke, B., Cossen, C., Forghani, B.,
`Osmond, D. Salivary Shedding Of KSHV Among Homosexual Men. Third Annual International
`HHV—8 Meeting. July 2000.
`Kaplan LD, Herndier B, Klencke B, Gascon R, Kesterson J, McGrath MS. Poyamine Analogue
`Therapy with Diethylhomosperrnine (DEHOP) for HIV-Associated non-Hodgkin's Lymphoma:
`Targeting Proliferating Macrophages. VII International Conference on Malignant Lymphoma 2-5
`June, 1999, Lugano Switzerland Abstract # 105 p 35
`Kaplan LK, Hemier B, Klencke B, Gascon R, Kesterson R, McGrath MS. Diethylhomospermine
`(DEHOP) for HIV-associated non-Hodgkin’s lymphoma (HIV-NHL). Third National AIDS
`Malignancies Conference May 26-27 1999, Abst 117.
`Benson AB, Mitchell E, Abramson N, Klencke B, Ritch P, Burnham J, McGuirt C, Levin J,
`Hohneker J. A multicenter, phase II trial of oral eniluracil plus 5 -FU in patients with inoperable
`hepatocellular carcinoma. PASCO 1999, Abstract 985.
`Chang IWC, Vaquerano JE, Cadbury P, Peng M, Zhou YM, Wong B, Kashani-Sabet M, Klencke
`BJ, Sagabiel RW, Ituarte P, Leong SPL. Active specific immunotherapy with autologous tumor
`vaccine plus recombinant GMCSF with and without interferon alfa—2B mediates tumor regression in
`stage IV melanoma patients. PASCO 1999, Abstract 1697.
`Blackbourn D., Lennette E., Witte M., Klencke B., Levy J. Cellular host range of human
`herpesvirus 8 (HHV8). Abstract, AACR meeting 1997.
`Klencke B, Soetikno R, Corkins S, Venook AP, Warren RS, Garber A. Hepatic Artery Infusion
`Pump Therapy for Colorectal Cancer: Is It Cost-Effective? Abstract, AFCR Clinical Research
`Meeting 1995
`Chapters
`
`Klencke B., Volberding P. AIDS-Related Malignancies. In: Holmes et a1. Sexually Transmitted
`Diseases, 3rd edition. New York: McGraw—Hill, 1998, Chapter 76.
`
`PHIGENIX
`
`Exhibit 1029-13
`
`

`

`‘ EXHIBIT B
`
`PHIGENIX
`
`xhibit 1029-14
`
`

`

`,fetal tissues
`
`© Macmillan Press Ltd,._199'0
`
`/ Oncogene (1990), 5, 953—962
`
`Expression of the HER-Z/neu prom-oncogene in normal human adult and-
`
`
`
`
`Michael F. Pressl, Carlos Cordon—Crude2 and Dennis J. Slamo-n3
`.
`.
`1Department ofPathology, UniversityofSouthern California, 201]
`_
`Pathology, MemorialSloaneKetr-eriny Cancer Center, 1275 York Avenue, New York, New York 10021; JDivision ofHematology
`and Oncology, Department ofMedicine, UCLA Medical School, Los Angeles, California 90024, USA
`
`The HER-Zlneu proto-oncogerle is homOlOgous With, but
`nodeenegat-ive' breast cancer (Wright et al., 1989; R0 et
`distinct
`from,
`the epidermal growth factor reCeptor.
`at, 1989; Paik et al., 1990); HER-2/n‘eu amplification
`Current evidence indicates, that this gene is frequently
`and overexpression have also been found in human
`’ amplified and/or overexpres'sed in some human breast
`ovarian carcinomas where the alteration is again as‘sOci—
`and ovarian cancers and that these alterations may be
`ated with a shorter overall survival
`(Slamo-n er al.,
`- clinically important; hawever, little is known about the
`19893, b). Amplification of this gene is consistently
`expression pattern of the gene in normal tissues. Using
`associated with overexpression of both the 4.5kb mes—
`immnnohistoc‘hemistry and northern blot analyses to
`senger RNA encoding the gene and the p185 protein
`identify the HER-Z/neu protein and transcript respec-
`product. Several lines of experimental evidence indicate
`tively, we have evaluated a variety of normal adult and
`that alterations in HER—Z/neu can play an important
`cells in the gastrointestinal, respiratory,
`reproductir‘ze,
`by the mouse mammary tumor virus promoter consis-
`and urinary tract as well as in the skin, breast and pla-
`tently develop breast carcinoma (Muller et al, 1988).
`center. Northern hybridization confirmed the presence of
`Transfection
`and
`subsequent
`amplification/
`the 4.5’kb transcript encoding the protein in these tissues.
`overexpre‘ss‘ion or overexpressi’on alone of a normal
`The amount ofHER-2/n‘e‘u message and protein was gen-
`HER—Z/neu gene (Hudziak at al, 1987; diFiorc er £21.,
`
`orally higher in fetal tissues than in the corresponding
`1987) renders NIH3T3 cells more tumorigenici'n nude
`normal adult
`tissues. HER-2/neu expression levels in
`mice. The HER-Z/neu gene encodes a membrane protein
`these normal tissues were similar to the levels found in
`with an extracellular domain, which makes it a candi—
`non—amplified, non-overexpre‘s‘sing breast cancers and
`date target for immunotherapeutic approaches against
`breast cancer
`cell
`lines. Southern hybridization of
`those cells expressing high levels of gene prOduct.
`extracted DNA showed that none of the normal tissues
`Experimental data using neu~transformed cell
`lines
`expressing HER-2km: had amplification of the gene.
`grown in vitro as well as in nude mice shggest that this
`These results confirm that HER-217ml
`is normally a
`type of approach may be feasible (Drebin at £21., 1985).
`membrane Constituent ofa variety ofepithelial cell types.
`To explmt
`these potential
`therapies in panents it 13
`important to understand the distribution and relative
`" "““h‘“ " '
`'
`expressiOn levels of HER~2/neu in normal tissues. Little
`'
`Introduction
`This study describes the distribution of HER—2/neu
`isolated from DNA
`first
`The neu oncogene was
`terizes the expression levels.
`extracted from ethylnitrosourea-induced adrenal neuro-
`glioblastomas of neonatal rats (Shih et al, 1981). The
`Results
`oncogene was identified by its ability to function as a
`dominant
`transforming gene in NH'I3T3 transfection
`HER—2/ncu expression, in both fetal and adult tissues,
`assays. Since this gene was identified in DNA from ~was identified immunohistochemically on the mem-
`neuro‘g‘lioblastomas it was referred to as neu (Schechter
`branes ofepithelial cells (Table 1, Figures 1 and 2). This
`er 0L, 1984) Subsequently, three different groups inde-
`staining pattern was found throughout
`the gastro-
`pende‘ntly identified the human 1101110108 Of this gene
`intestinal, respiratory, urinary, and reproductive tracts,
`(COUSSC'IIS 873' 0L, 1985; Semba 91‘
`‘11-, 1985; King, KIaUS
`as well as the skin of fetal and adult specimens. Levels
`& Aaronson, 1985) and, because of homology t0 the
`of expression were, in almost all instances, higher in,
`human epidermal grthh factor receptor (HER) and
`fetal than in the cOrresponding adult tissues. Immuno—
`c~erb~B prom-oncogene, it was referred to as HER-2
`staining of frozen, normal tissues ranged from barely
`(0011535115 er 01-: 1985.) 01” c~erb-B-2 (Sern'bga 9‘ al., 1985)-
`detectable to,
`in a few specimens, moderate intensity
`Recently, amplification and overexpression ofthe HER-
`(2+)
`(Table
`1').
`lmmunostaining in formalin~fixed,
`2/neu gene was found to be associated with shortened
`paraffin—embedded tissues ‘was also seen on cell mem-
`. disease-free and overall survival in women With DOdC'
`branes; however, these sectionscOnsistently showed less
`DOSitiVC breast cancer (Slamon 9t 611-7 1937; 1989a, b)- In
`immunostaining than the corresponding frozen sections
`addi