Ac1({Amm'canAssociationfor cancerkesearch
`
`
`
`Anhual Meeting
`
`Ajm'Z 6-10, 2002 - San Francisco, California
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`i/blume 43 - March 2002
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`Injoint sponsorship with the Keck School of Medicine of the University of Southern California
`
`The p:'eim'er :23:-?€tii:g_f’or
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`;“(’.'\.‘(’(U‘("h in the po.s'f-g'ei:r_>:r1fc'- era
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`-i'EALTH SCIENCES i.iE5RARY
`UNa'L-'ERS.1TY 0:: WISCONSIN
`
`a‘-..-ma
`
`:4
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`3:332
`
`F305 Lind en Drive
`Madison, WI 53TOEi
`
`West-Ward Pharm.
`Exhibit 1030
`Page 001
`
`West-Ward Pharm.
`Exhibit 1030
`Page 001
`
`

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`AMERICAN ASSOCIATION FOR CANCER RESEARCH, INC.
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`Officers
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`President
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`. ..................W/aunK.il-long
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`President—Elect
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`. .Susan Band I-Iorwitz
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`Treasurer .
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`. Bayard D. Clarkson
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`. .Tom Curran
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`Past President
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`Chief Executive Officer .
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`. .Margaret Foti
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`Until 2002
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`Mina]. Bissell
`Michael B. Kastan
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`Edison T. Liu
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`Frank Rauscher, III
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`Board of Directors
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`Until 2003
`Michaele C. Christian
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`Lorraine]. Gudas
`Willimn G. Nelson, V
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`Peter K. Vogt
`Geoffrey M. Wald
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`Until 2004
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`Frederick R. Appelbaurn
`Ronald A. DePinho
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`Tyler E. Jacks
`George F. Vande Woude
`Barbara L. Weber
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`Address inquiries to the Olilice of the American Association for Cancer Research, Inc. (AACR). Public Ledger Building. Suite 826, 150 5.
`Independence Mall Wem, Philadelphia, PA 19106-3483 [Telephone: (215) 440-9300; Fax: (215) 440-9313].
`
`The Prueeedirrgs afsbe American Anwiancnfir Canter Remand?) is printed for the AACR by Cadmus Professional Communications, Linthicum, MD
`21090-2908 and is included in the membership dues For active and associate members. Volume 43 of the Prcrmiirigs oftlie American A:.rcer‘an'onfi-r
`Cancer Res-em-:3: (ISSN 019?—016X) succeeds Volume 42 of the Pmceedingr oft!» Anuriiran .A.ooc:'ao‘or:fiar Clearer Remsrtb. The B-oceedings may be
`obtained at a price of $55.00 through registration at the Annual Meeting of the American Association For Cancer Research, April 6-10, 2002, or
`ordered by writing to: AACR Subscription Oflice, I-'20. Box 11806, Birmingham, AL 35202 [Telephone: (300) 533-4931 or (205) 995-1567; Fax:
`(205) 995-1583]. Add $6.00 For shipping for orders outside the US.; expedited delivery rates are available on request.
`
`The ficeudbrgt qfrbe American Auaeiafionfir (hum Restart}: is copyrighted © 2002 by the AACR. All rights reserved. Redistribution or resale of
`the Boceedings or ofany materials in the Proteedings. whether in machine readable, other electronic. or any other form, is prohibited. Reproduction
`for advertising or promotional purpocm. or republication in any Form, may be permitted only under license from dte AACR. Any reproduction,
`whether electronic or otherwise, of abstracts beyond that permitted by copyright law must be authorized in writing in advance by the AACR.
`Requests to reproduce abstracts will be considered on an individual basis and permission may be granted contingent upon payment of an appropri-
`ate fee. Reproduction requests must include a briefdescription of intended use. Third parties should also obtain the approval of the authors before
`corresponding with the AACR. Failure to oomply with the foregoing restrictions and unauthorized duplication of any portion of these materials are
`a violation of applicable laws and may be subject to criminal prosecution and civil penalties.
`
`is accepted by the Editors, by the American Association lint Cancer Research, Inc., or by Cadmus Professional
`No
`Communications fin the opinions expressed by the contributors herein.
`
`_/$HC{
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`93rd Annual Meeting
`
`West-Ward Pharm.
`Exhibit 1030
`Page 002
`
`West-Ward Pharm.
`Exhibit 1030
`Page 002
`
`

`

`differentiation of myeloid leukemia cell lines (NB4. HL—60 and U937’). 85 was
`identified in our recent studies as a potent inhibitor of PTPases. Herein. we
`present data demonstrating that SS (250 pgiml. 6 days} induced S?% of NB4
`cells to reduce nitroblue tetrazoiium (NED. in comparison to the 90% induced by
`_,gTFiA (‘f
`_i.l.M. 6 days}. SS-induced NB4 cell differentiation was confirmed by
`increased CD11b expression and asmiated with growth arrest at S phase and
`increased cell death. Our results showed further that SS-induced NB4 differen-
`iiation was irreversible and required continuous drug exposure for optimal induc-
`tion. Moreover. SS (400 pgiml, 6 days} induced 60% and 55% of NBT-positive
`cells in HL—50 and U937 cell lines. which were augmented in the presence of
`GM-CSF (2 ngimi) to levels (85% and 81%. respectively] comparable to those
`induced by ATFIA. These results provide the first evidence of a differentiation
`induction activity of PTPase inhibitor SS in myeloid leukemia cell
`lines and
`gsuggest its potential therapeutic use in myeloid leukemia. Since 33 induces
`differentiation via targeting F'TPases. a mechanism distinct from that of ATRA. it
`may be particularly useful in Ali.-'lL cases unresponsive or developed resistance to
`ATRA.
`
`#356 NAD(P]H:quinone oxidoreductase iivoo-ii-ciepenctant and -indepen-
`dent cytotoxlcity of potent quinone cdc25 phosphatase inhibitors. Yusheng
`Han. Hongmei Shen. Brian Carr, John S. Lazo, and Su—Shu Pan. Uni'versi‘ty of
`Pi'ti'Sburg'h Cancer institute, Pittsburgh. PA. and University of Pittsburgh. Pitts-
`burgh. PA.
`A vitamin K analogue. compound 5 (Ci:.ld5. a thioethanol naphthoquinone),
`inhibits onoogenic Cdc25 phosphatases, and arrests cell cycle progression at
`both (31 and ozmi. Flecently. we evaluated >10.000 compounds in the NCl
`chemical repository for in vitro inhibition against recombinant human Cdc25B
`phosphatase and identified a ciuinone substructure in many of the active com-
`pounds. Bioreductive enzymes in cells. however. are known to reduce various
`quinones resulting in either detoxification or activation. Therefore. we used an
`isogenic set of human colon cancer cell lines to evaluate the effect of N001 on
`the cytotoxic activity of Cpd5 and the two most potent phosphatase inhibitors
`from the repository: NSC 9539? (a bis-thioethanol naphthoquinone) and N30
`663284 (a ouinolinedione). The two cancer cell lines used were HCT11E, which
`has intermediate N001 activity. and its mitomycin C—resistant sub-line HCT116—
`Fl30A (Fi30A}. which has minimum NCJO1 activity. Cell survival was measured by
`colony formation after 7 days drug exposure. Cell cycle arrest was evaiuated by
`flow cytometry after 6 hr drug exposure. Cpd5 had an I659 of 2210.3 i..tl'v'l tor
`HCT1f6 and 0.23:'.0.05 i.iM for R30A.
`i.e. a 10-fold difference. Inclusion of
`dicoumarol (10 .i.tM). an inhibitor of N001, decreased the |C5o of Cpd5 for
`HCT116 to 024770.04 prvi, but had no effect on Fl30A. In contrast, HCT116 and
`Fi3ClA cells were equally inhibited by NSC 953?5 with lC5ns of 1.4:0.3 i.i.ll.-‘l and
`1.3:0.2 _ul\il. respectively. Similarly, HCT116 and HSOA cells were equally inhib-
`ited by NSC 653234 with lC5os of 2.41.0.3 _ul\.Il and 2.5105 p.l'v'l. respectively. All
`three compounds blocked the two cell lines at the G2i'M phase transition. con-
`sistent with odc25 inhibition. Cpd5 at 2.5 pM arrested Fi30A cells at G2iM but ?.5
`i.iM Cpd5 was needed to arrest HCT116 cells to a similar degree. NSC 953T5 and
`653284 arrested cell cycle progression at G2ilyl of HCT116 and Fi30A cells
`similarly, and did so in a concentration-dependent manner between 2.5 and ?.5
`;.lM. Our data imply that NOO1 in HCT116 cells protected cells from the action of
`Cpdfi. probably by the reduction of Cpd5 to a less active hydroxylquinone.
`In
`contrast. both NSC 9539? and 663284 displayed cytctoxicity that was indepen-
`dent of N001 levels. (Support: NCI CA61862 and CATSDSQ)
`
`#357 Antitumor and anticarcinogenic action of Cpd 5: A new class of
`Protein phosphatase inhibitor. Siddhartha Kar, Meifang Wang. Zhenggang Fien.
`Xiangbai Chen. and Brian I. Carr. University of Pittsburgh, Pittsburgh. PA.
`Background: We have chemically synthesized a new class of inhibitors of dual
`specificity phosphatases (DSP}. which play an important role in cell cycle and
`signal transduction. Cpd 5 or 2-(2-mercaptoethanol)-3-methyl-1 ,r1—naphthoqui—
`none is one of the most potent. It inhibits DSPs (especially the Cdc25 family} in
`tissue culture cells and induces tyrosine phosphorylation of various DSP sub-
`strates, including Colts and inhibits cell growth both in vifro and in vivo (JBC
`2?lJ:2B304, 1995; Proc. AACFI 39:224. 1998). Purpose: in this study we evaluated
`la! the antitumor and (b} the anticarcinogenic activity of Cpd 5 for the first time.
`Methods: (a,l JM1 hepatomas were grown in 2 month old Fischer male rats by
`Subcutaneous injection on the back or intra—portally in the liver. Rats were treated
`with Cpd 5 by intratumor. subcutaneous (nearby site}. intramuscular (distant site),
`01' intraperitoneal injection. either as a single high acute dose or chronically as
`Several low doses. lb) Flats were injected intraperitoneally with a single dose of the
`carcinogen N-Nitrosodiethylamine (DEN). lmrnunostained liver sections for gluta-
`Ihione-S—transferase—pi (GST-on detected pre—neoplastic foci after 3 weeks. Cpd
`5_ was injected subcutaneously or intraperitoneally two weeks after DEN as a
`Sfngle high acute dose or chronically as several low doses. Flesults: (a) Cpd 5 had
`5'Elnificant inhibitory effect on both intrahepatic (14% of control. p<0.00000008}
`and subcutaneous (33% of control. p<0.0000Bi tumor growth and also had
`Significant inhibitory effect when injected intramuscularly at a site distant from the
`"Jmor (50% of control, p<:0.002). There was no significant difference between the
`Effects after acute or chronic injections. However. toxicity was much lower with
`chronic treatment. (b) The number of enzyme altered foci was also significantly
`
`EXPERIMENTAUMOLECULAR THERAPEUTICS 3
`
`reduced when rats were treated with acute (40% of control, p<0.0-0002) or
`chronic (50% of control, p<o.02]i Cpd 5. Conclusions: Cpd 5 had significant
`inhibitory effect on growth of tumors and foci.
`
`#353 Bosentan, a novel endothelin-A and -B receptor antagonist inhibits
`proliferation of malignant melanoma cells. Alel-tsandar Sekulic, Padma Suresh.
`Mark Fl. Pittelkow, and Svetomir N. Ivlarkovio. Mayo Foundation. Rochester, MN.
`Here we tested a feasibility of endothelin (ET) receptor blockade with a due:
`endothelin-A and —B receptor (l:‘rFl-A and ETR—B) antagonist, Bosentan, as a
`novel therapeutic approach for malignant melanoma. ETs are 21aa peptides
`primarily produced by endothelial cells and implicated in a variety of physiological
`functions. Binding of ET to ETFi—A on vascular structures potently stimulates
`angiogenesis and.
`thus.
`likely plays an important role in growth of multiple
`cancers. Activation of ETFt—Bs. among other things. regulates melanocyte devel-
`opment and function. We first examined patterns of ErFl subtype expression on
`six established melanoma cell lines using flow cytometry and immunocytochem—
`istry. Following this sections of primary and metastatic melanoma tissues were
`evaluated for El'FlNETFiB expression by immunohistochemistry. To test func-
`tional effects Bosentan on melanoma cell proliferation. six melanoma cell lines
`were subjected to standard SH-thymidine incorporation assays in presence or
`absence of various concentrations of Bosentan. All examined melanoma tissues
`(2 primary. 9 metastatic} express ETFIB. albeit to different levels, whereas EFHA
`was expressed to low levels in only 3 metastatic tumors.
`In functional assays
`Bosentan inhibited proliferation of all examined cell lines with the I050 ranging
`between 7 and 40 pgrml. Our results suggest that malignant melanocytes express
`functional Eff-ls, and their treatment with Bosentan leads to significant growth
`inhibition. Concurrent inhibition of ETR-A and ETR—Bin vivo by low toxicity. orally
`available inhibitor Bosentan might therefore prove useful as a novel mode of
`anti—meIanoma therapy through simultaneous inhibition of cancer cell growth and
`process of angiogenesis.
`
`#359 in vivo activity of RADGO1. an orally active raparnycin derivative. in
`experimental tumor models. Terence O'Fteilly, Juliane vaxelaire, Melanie Muller.
`Heinz-Herbert Fiebig. Marc Hattenberger. and Heidi A. Lane. Business Unit On-
`cology. Novartis Pna.-ma AG. Basel. Switzerfand. and Oncofest grnbl-l, Freiberg.
`Germany.
`is a hydroxyethyl ether derivative of rapamycin that is orally bioavaiI—
`FlAD00t
`able. RAD001 has demonstrated in vitro anti-proliferative activity against a panel
`of human tumor lines. For in vivo testing. tumor-bearing nude mice were admin-
`istered FIAD-001 in a variety of doses and schedules. Tumors were established by
`transplantation of fragments generated from injection of cells, or by transplanta-
`tion of fragments from stabilized tumors originating from surgically removed
`human tumors. when administered once daily p.o., at doses ranging from 0.5-5.0
`mg/kgiday. RAD001 was a potent inhibitor of tumor growth in 10 different
`xenograft models of human tumors (including pancreatic, colon. epidermoid. lung
`and melanoma).
`Ir! general, RAD001 was well tolerated and better tolerated in
`mouse xencgrafl models than standard cytotoxic agents {i.e. doxorubicin and
`5—fluorouraciii. while possessing similar antitumor activity. Only one instance of in
`vivo resistance has been observed (MAXF 401 mammary xenograft model].
`otherwise the activity of HAD001 was generally inhibition of tumor growth {per-
`sistent regressions in one tumor line. TIC values of 9 to 45 % in 8 tumor lines).
`Xenograft models sensitive to RAD001 treatment included tumors exhibiting
`comparative resistance in vitro {KB-31 and HCT116). Persistent tumor regres-
`sions (41 %} were observed in a line displaying sensitivity to RAD001 in vilro
`(A5491. Pharmacokinetic analyses. following a 5 mgfkg administration, indicated
`rapid uptake into plasma (Cmax 2513 ng/ml; Tmax 1 h). but the time to Cmax was
`delayed in tumors (Cmax 102 ng/g; Tmax 2 hi. Elimination from the tumor {tti’2,
`16 hr) was apparently slower than for plasma (H12. ?.5 hr}. RAD001 levels were
`above the I050 of A549 cells for a '32 h period. Interestingly, tumor RAD-O01 levels.
`following a single 5 mgikg administration. never exceeded the in vitro antiprolif-
`eralive I050 for either KB-31 or HCT116 cells: despite the sensitivity of these lines
`in vivo. From these observations. and given the extreme sensitivity of endothelial
`cells to RAD001. it is plausable that HAD-001 may not only act on tumor cells but
`may also affect angiogenesis. Taken together, these data support the application
`of FlAD(}0l as an antitumor agent.
`
`3360 Discovery of anticancer agents from sponge-associated fungi. Fred-
`erick A. valeriote. Karen Tenney. Charles Grieshaber, Halina Pietraszkiewicz.
`Akiko Amagata. Taro Amagata. Jeff Gautschi. Joseph Media. Joseph Stayanofi.
`Flichard Wiegand. and Phil Crews. Henry Fond Health System. Detroit. Ml, and
`University of California Santa Cruz, Santa Cruz. CA.
`We have evaluated 1.112 extracts (from 560 spongeassociated fungi} for
`assessment of potential anticancer activity. Both broth and mycelia extracts were
`assayed in most cases. Each sample was assayed in vitro against up to 8 cell
`types (murine and human) in a disk diffusion! clcnogenic assay. From these
`results. the samples were assigned into one of 4 categories: Inactive (79% of the
`extracts}. Equally active across cell types (16% of the extracts). Equally active and
`potent (9 extracts or 1%), and Solid tumor selective (42 extracts or 3.8%). The
`equally active and potent category is studied further since solid tumor selective
`compounds might exist in the extract but be concealed by one or more potent.
`cytotoxic compounds. Further, a novel. potent compound could form the basis
`for analog synthesis in an attempt to develop an active anticancer agent. One
`
`Proceedings of the American Association for Cancer Research 0 volume 43 0 March 2002
`
`?1
`
`This material may be protected by Copyright law (Title 17 U.S. Code)
`
`West-Ward Pharm.
`Exhibit 1030
`Page 003
`
`West-Ward Pharm.
`Exhibit 1030
`Page 003
`
`