PROCEEDINGS
`
`EIGHTY-EIGHTH ANNUAL MEETING
`
`APRIL 12- I6, I997
`
`SAN DIEGO, CA
`
`VOLUME 38 I MARCH I997
`
`OSI EXHIBIT 2006
`APOTEX V. OSI
`|PR2016-01284
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`i
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`OSI EXHIBIT 2006
`APOTEX V. OSI
`IPR2016-01284
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`
`teenock Memoria ”-
`University of Wisconsin - Madison
`550 Babcock Drive
`Madison. WI 5370§flfiE-EHCAN AS
`
`PROCEEDINGS OF THE
`SOCIATION FOR CANCER RESEARCH
`
`Executive Director and
`Director of Publications
`Margaret Foti
`Publications Staff
`Associate Director
`of Publications
`Mary Anne Mennite
`
`Manager, Editorial Services
`Heide M. Pusztay
`Staff Editors
`Michael J. Beveridge
`Kathleen C. Assenmacher
`Pamela R. Arnold
`Laurie B. Baker
`Ronald M. Vitale
`
`Staff Assistant
`Mary Ellen Pirring
`Administrative Coordinator
`Holly M. Haskins
`
`Senior Editorial Assistant
`Theresa A. Griffith
`
`Editorial Assistants
`Valerie L. Samuel
`Andrea Conrad
`Cecilia Gallo
`Brenda Roberson
`
`Marketing Coordinator
`Robert B. O‘Malley
`
`AMERICAN ASSOCIATION FOR CANCER RESEARCH, INC.
`Administrative Staff
`Director of Administration
`Adam D. Blistein
`Controller
`Joan D. Ritchie
`
`Officers
`
`Louise C. Strong, President
`Donald S. Coffey, President-E/ect
`Bayard D. Clarkson, Treasurer
`Joseph R. Bertino, Immediate Past President
`Edward Bresnick, Past President
`Margaret Foti, Executive Director
`
`coordinator, Financial Operations
`George L. Moore
`
`Manager, Meetings and Exhibits
`Jeffrey M. Ruben
`
`Meeting Planner
`Carole L. Kanoff
`
`Public Information Coordinator
`Jenny Anne Horst—Martz
`
`Assistant to the Executive Director
`Ruth E. Fortson
`
`Membership Development Coordinator
`Robin E. Felder
`
`Administrative Assistant
`Margaret A. Pickels
`
`Systems Specialist
`Lydia l. Rodriguez
`Accountant l
`Monica Storione
`
`Staff Assistants
`Jennifer S. Roberts
`Maria M. Edstrom
`
`Meetings Assistant
`Jinhui Yun
`
`Financial Assistant
`Jeriesha L. Williams
`
`Secretary
`Malika L. Wright
`Editorial Assistant
`Diana F. Certo
`
`Office Clerk
`James J. Waters
`
`Data Entry Clerk
`Robert A. Simms ll
`
`Board of Directors
`
`Term Expiring 1997
`.Webster K. Cavenee
`Philip C. Hanawalt
`Stanley J. Korsmeyer
`Thea D. Tlsty
`
`Term Expiring 1998
`Clara Derber Bloomfield
`Michael M. Gottesman
`Ann R. Kennedy
`Frederick P. Li
`
`Term Expiring 1999
`Susan P.C. Cole
`Eric R. Fearon
`Stephen H. Friend
`Waun Ki Hong
`
` _j
`Address inquiries to the Office of the American Association for Cancer Research, Inc. (AACR), Public Ledger Building, Suite 826, 150 S. independence Mall West,
`Philadelphia, PA 191063483 [Telephone: (215) 440-9300; FAX: (215) 440-9313].
`
`y Cadmus Journal Services, Linlhicum, MD 21090-2908 and included in
`esearch, Cell Growth & Differentiation, Cancer Epidemic/og Biomarkers & Prevention, and Clinical Cancer Research
`
`No responsibility is accept
`expressed by the contributor
`
`comply with the foregoing restrictions and
`ay be subject to criminal prosecution and civil penalties.
`
`ii
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`This material may be pmtected by Copyright law (Title 17 us. Code)
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`PHARMACOLOGY/THERAPEUTICS (PRECLINICAL AND CLINICAL) 44
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`(CD31 staining) in CGP 53716 treated with mice. Collectively, these data suggest
`that inhibition of tumor stroma formation and angiogenesis following CGP 53716
`treatment results in growth inhibition of H226 carcinoma in the lung.
`
`#4246 CGP 57148B, a protein-tyrosine kinase inhibitor with potential for
`the treatment of Bcr-Abl positive leukemias and diseases involving deregu-
`lation of PDGF receptor and c-Kit tyrosine kinases. Buchdunger, E., Zimmer-
`mann. Jt, Mett, H., Muller. M.. Law, N., Cioffi, C., Druker, B., and Lydon, N. Ciba
`Pharmaceuticals Division, Oncology Research Department, Ciba-Geigy Ltd., CH-
`4oo2 Basel, Switzerland, Ciba Pharmaceuticals Division, Ciba-Geigy Ltd., Sum-
`mit, New Jersey 07901, Oregon Health Sciences University, Portland, Oregon
`201
`97cGP 57148B has been identified as a potent protein—tyrosine kinase inhibitor
`with selectivity for the Abl and PDGF receptor tyrosine kinases. Cellular prolifer-
`ation and tumor growth of Bcr-Abl or v-Abl expressing cells were specifically
`inhibited by this compound. CGP 57148B selectively inhibited PDGF—mediated
`cellular events such as PDGF receptor autophosphorylation, inositol phosphate
`formation and c—fos induction. The compound was a potent inhibitor of PDGF-
`mediated growth of v—sis transformed BALB/c 3T3 cells and PDGFstimu|ated
`proliferation of A10 rat aortic smooth muscle cells. In vivo, CGP 57148B inhibited
`growth of a number of PDGF—dependent tumors. Combination of CGP 57148B
`with cytotoxic agents resulted in tumor regression and cures, with different
`xenografts being sensitive to different drug combinations. Amongst Type III
`receptor tyrosine kinases, CGP 57148B was found to potently inhibit the SCF
`receptor, c-Kit, but not
`the related c-Fms, F|t—3 and Flt—1
`tyrosine kinases.
`Increasing evidence suggests that inappropriate receptor signaling by c-Kit may
`be involved in a number of cancers,
`including small cell
`lung cancer (SCLC).
`Interestingly, tumor growth of NCl—H69 and NC|—H209 SCLC lines was sensitive
`to inhibition by CGP 57148B which is compatible with the inhibition of the c-Kit
`tyrosine kinase. Taken together, these results suggest that CGP 57148B is a
`signal transduction inhibitor which might have therapeutic potential for the treat-
`ment of Ph+ leukemias as well as diseases which involve abnormal activation of
`PDGF receptor and c-Kit tyrosine kinases.
`
`#4247 Cytoxicity and phosphotyrosine effects of c-src kinase inhibition
`by substituted pyridopyrimidine tyrosine kinase inhibitors in human colon
`carcinoma cell lines. Kraker, A.J., Moore, C.W., Amar, A.M., Shen, C.S., Nelson,
`J., Slintak, V., Fry, D.W., Lu, G., Panek, R., Klutchko, 8., and Hamby, J. Parke-
`Davis Pharm Res. Div. of Warner—Lambert Co., Ann Arbor, MI 48105
`Tyrosine kinase inhibition has been identified as a potential therapeutic strategy
`in neoplastic disease. Potent c-src kinase inhibitors (IC5,,=9nM in isolated c-src
`assay for 6-(2,6-dich|oro—pheny|)—2—[4—(2—diethy|amino-ethoxy)-phenylamino]—8H-
`pyrido[2,3—d]pyrimidin—7—one dihydrochloride)(PD166285, A) are described. Src
`family members fyn,
`lyn, and lck are also potently inhibited in vitro. A causes
`growth delay on plastic with IC5o values of 140nM for HT—29, 426nM for HCT—8
`and 270nM for SW-620 cell
`lines.
`In soft agar clonogenic assays with 2 day
`treatment followed by drug removal, A gave IC50 values of 1.8 [.LNl(HT-29),
`780nM(HCT—8), and 330nM(SW—620). Phosphotyrosine(P-Y) content of focal ad-
`hesion kinase(FAK) and paxi|lin(known substrates of c-src) is reduced at SUD-p_M
`concentrations in HT-29 cells treated with A for 2 hours and then mitogenically
`stimulated. In a fibroblast cell line overexpressing epidermal growth factor recep-
`tor(EGFr) and c-src, A is a specific c-src inhibitor(reducing FAK and c-src P-Y)
`compared to EGFr kinase activity(receptor P-Y). Because of potent,kinase inhi-
`bition. selectivity, and cellular signaling effects of these compounds,
`in vivo
`studies are underway.
`
`#4248 CP—358,774: A selective EGFR kinase inhibitor with potent antipro—
`liferative activity against HN5 head and neck tumor cells. lwata, K., Miller,
`P.E., Barbacci, E.G., Arnold, L., Doty, J., DiOrio, C.l., Pustilnik, L.R., Reynolds, M..
`Thelemann, A., Sloan, D., and Moyer, J.D. Oncogene Science Inc., Uniondale, NY
`11553-3649, Pfizer Central Research, Groton, CT 06340
`CP—358,774 is an inhibitor of the epidermal growth factor receptor (EGFR)
`tyrosine kinase (TK), an activity which is overexpressed in many carcinomas.
`CP—358,774 inhibits isolated human EGFR TK with an I050 of 2 nM, and reduces
`EGFR autophosphorylation in MDA—MB—468 tumor cells in culture with an |C50 of
`20 nM. EGFR TK is >1000—fo|d more sensitive to CP—358,774 than other TKs such
`as human c-src, insulin receptor, insulin—like growth factor receptor,or v—abl. As a
`further indication of selectivity, EGF-induced tyrosine phosphorylation of SHC
`proteins in HN5 cells is completely blocked by CP—358,774, whereas the insulin-
`Induced phosphorylation of IRS—1 is unaffected. CP—358,774 inhibits EGF—stim—
`Ulated mitogenesis in Fischer rat embryo cells with an |C5D of 70 nM, but does not
`similarly inhibit mitogenesis stimulated by platelet derived growth factor, insulin-
`Iike growth factor, or basic fibroblast growth factor, which also act through
`transmembrane receptors with TK activity. The proliferation of HN5 cells, which
`overexpress EGFR, is inhibited by CP—358,774 at 50 nM, and completely blocked
`at 250 nM. Pretreatment of athymic mice with CP—358,774 (100 mg/kg) com-
`Dletely inhibits EGF-induced autophosphorylation of human EGFR in HN5 tumor
`Xenografls and of EGFR in mouse liver. This inhibitor has potential for the
`treatment of tumors that are dependent on the EGFR pathway for proliferation.
`
`#4249 Therapy of human carcinomas in athymic mice by inhibition of EGF
`receptor-mediated signal transduction with CP-358774: Dynamics of recep-
`tor inhibition and anti-tumor effects. Pollack, V.A., Savage, D.M., Baker, D.A.,
`Tsaparikos, K.E., Sloan, D.E., Barbacci, E.G., Pustilnik, L.R., Smolarek, T.A.,
`Davis, J.A., Vaidya, M.P., and lwata, K. Dept. of Cancer, Pfizer Central Research,
`Groton, CT 06340, Oncogene Science, lnc., Uniondale, NY 11553
`Tyrosine phosphorylation of EGF receptors is an important early event in signal
`transduction and tumor cell replication. We devised an ex vivo assay to quantitate
`EGFr-specific tyrosine phosphorylation in human tumors obtained as s.c. xeno-
`grafts in athymic mice. Using an enzyme-linked immunosorbent assay (ELISA),
`we observed reproducible ED50‘s and have used this assay to describe the extent
`and duration of drug action in vivo. CP—358774 is an effective, orally active
`inhibitor of EGFr tyrosine phosphorylation (ED50= 10 mg/kg po). It has significant
`duration of action, producing, on average, 70% inhibition of EGFr-PY over a 24—hr
`period. Most importantly, we observed that inhibition of EGFr-PY in an ex vivo
`assay effectively correlates with the potency and degree of inhibition of EGFr-
`dependent human HN5 tumor growth in a xenograft therapy model (ED50: 10
`mg/kg qd X 5). These data suggest that CP-358774 may be an important new
`agent for therapy of EGFr-overexpressing human cancers.
`
`#4250 Pyrido[d]pyrimidine inhibitors of the tyrosine kinase activity of the
`EGF receptor: A binding model and structure-activity relationships for sol-
`uble analogues. Denny, W.A., Palmer, B.D., Flewcastle, G.W., Thompson, A.M.,
`Bridges, A.J., Doherty, A.M., Fry, D.W., Nelson, J.M., Rubin, J.R., Showalter,
`H.D.H., and Trumpp—KaI|meyer, S. Cancer Society Res. Lab, University of Auck-
`land, Private Bag 92019, Auckland, New Zealand, Parke—Davis Pharmaceutical
`Res., Division of Warner—Lambert Co., 2800 Plymouth Rd., Ann Arbor, MI 48105
`4-Anilinopyridoldlpyrimidines are potent, selective inhibitors of the tyrosine
`kinase activity of the epidermal growth factor receptor (EGFR), binding at the ATP
`site with ICE-,0 values down to 8 pM. They are of interest as anticancer drugs, since
`EGFR is over—expressed in many cancers and associated with poor clinical
`prognosis. A model for their binding to the EGFR was constructed from the
`catalytic subunit of the related cAMP—dependent protein kinase, and from struc-
`ture-activity data. This attributes their high selectivity to binding of the 4—ani|ino
`ring in a hydrophobic pocket of composition unique to the EGFR. The 6- and
`7-positions of the pyridoidlpyrimidines occupy the entrance of the ATP binding
`pocket, with the 7—position pointing towards the rlbose binding site of ATP.
`In
`agreement with this, analogues with weakly basic solubilising groups at the 6- or
`7—positions retain potent inhibitory activity towards EGFR. They are also potent
`inhibitors of EGFR auto—phosphory|ation in A431 cells. Examples evaluated
`against murine colon 38 tumors and human tumor xenografts in mice gave
`substantial growth delays.
`
`#4251 ZD1839, an epidermal growth factor tyrosine kinase inhibitor se-
`lected for clinical development. Woodburn, J.R., Barker, A.J., Gibson, K.H.,
`Ashton, S.E., Wakeling, A.E., Curry, B.J., Scarlett, L., and Henthorn, L.R. Zeneca
`Pharmacueticals, Alderley Park, Macclesfield, Cheshire, U.K. SK10 4TG
`Epidermal growth factor receptor (EGFR) is overexpressed in a wide variety of
`solid human cancers including non-small cell lung, breast, head and neck, blad-
`der and ovarian carcinomas. ZD1839, an anilinoquinazoline, is a potent inhibitor
`(0.02 micromolar) of the EGFR tyrosine kinase in vitro and of the EGF-stimulated
`growth of KB oral carcinoma cells in culture (0.08 micromolar). ZD1839 has
`excellent oral bioavailability and shows antitumor activity in a broad range of
`human solid tumor xenografts implanted in nude mice in the dose range 12.5 —
`200 mg/kg once per day orally. Sensitive tumors include A431 vulval, KB oral,
`A549 NSCLC, DU145 prostate, HT29, HCT15, CR10 and LoVo colorectal, and
`HX62 ovarian. Antitumor effects range from reduced growth rate to stasis; with
`marked regressions seen in some tumors. Therapy for up 4 months in nude mice
`is well tolerated. On the basis of these exciting pre—c|inica| data ZD1839 has been
`selected for clinical development.
`
`#4252 Novel benzoylacetylenic compounds: Potent and selective EGF
`receptor tyrosine kinase inhibitors. Suzuki, T., Kitano, Y., Ohya, J., Umeki, H.,
`lnokawa, H., Kawahara, E., Nakamura, H., Takayanagi, H., and Hara, H. Pharma-
`ceuticals Lab., Yokohama Research Center, Mitsubishi Chemical Co., Yokohama,
`Japan
`Inhibition of oncogenic tyrosine kinases is a potential approach for the treat-
`ment of cancer. We have synthesized a novel series of benzoyl acetylenic corn-
`pounds as potent and selective inhibitors of EGF receptor tyrosine kinase. We
`found that one of these compounds, DAB—720, has excellent biological activities
`in in vitro and in vivo studies. We show here the structure—activity relationships of
`these derivatives and biological profiles of DAB-720. Using a panel of protein
`kinases including EGF receptor, PDGF receptor, c-src, lck and PKC, DAB—72O
`displayed selective inhibitory activity against EGF receptor with ICSD of 0.070 iLM
`and little or no inhibition for the other kinases at 50 piM, except lck for which
`DAB-720 had an |C5o of 16 pM. Autophosphoiylation of EGF receptor in A431
`cells and of HER2/Erb B2 in BT474 cells were also inhibited over 50% at 5 pM and
`10 ,iM respectively. The |C50 for KB cell growth was 0.76 pM. In in vivo study
`significant growth inhibition of KB xenograft in nude mice was demonstrated by
`30 mg/kg intraperitoneal administrations of DAB—720.
`
`Proceedings of the American Association for Cancer Research 0 Volume 38 0 March 1997
`
`638