PROCEEDINGS
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`EIGHTY-EIGHTH ANNUAL MEETING
`
`
`
`APRIL 12-16, 1997
`SAN DIEGO, CA
`VOLUME 38 ® MARCH 1997
`
`i
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`OSI EXHIBIT 2006
`APOTEX V. OSI
`IPR2016-01284
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`i
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`OSI EXHIBIT 2006
`APOTEX V. OSI
`IPR2016-01284
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`

`Steenbock Memorial Library
`University of Wisconsin - Madison
`PROCEEDINGS OF THE
`990 Babcock Drive
`Madison, WI 5°370¢sMERICAN ASSOCIATION FOR CANCER RESEARCH
`
`Executive Director and
`Director of Publications
`MargaretFoti
`Publications Staff
`Associate Director
`of Publications
`Mary Anne Mennite
`
`Manager, Editorial Services
`Heide M. Pusztay
`Staff Editors
`MichaelJ. 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
`TheresaA.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
`JoanD.Ritchie
`
`Officers
`
`Louise C. Strong, President
`Donald S. Coffey, President-Elect
`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
`MargaretA. Pickels
`
`Systems Specialist
`Lydia |. Rodriguez
`Accountant|
`Monica Storione
`
`Staff Assistants
`Jennifer S. Roberts
`Maria M. Edstrom
`
`MeetingsAssistant
`Jinhui Yun
`
`Financial Assistant
`Jeriesha L. Williams
`
`Secretary
`Malika L. Wright
`Editorial Assistant
`Diana F. Certo
`
`Office Clerk
`JamesJ. Waters
`
`Data Entry Clerk
`Robert A. SimmsII
`
`Board of Directors
`
`Term Expiring 1997
`Webster K. Cavenee
`Philip C. Hanawalt
`Stanley J. Korsmeyer
`TheaD. 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
`
`
`:ee
`Addressinquiries to the Office of the
`(AACR), Public Ledger Building, Suite 826, 150 S. Independence Mall West,
`American Association for Cancer Research,Inc.
`Philadelphia, PA 19106-3483 [Telephon
`e: (215) 440-9300; FAX: (215) 440-9313].
`The Proceedingsof the American Association fo
`r Cancer Researchis printed for the AACR by CadmusJournal Services, Linthicum, MD 21090-2908andincludedin
`membersubscriptions to the journals Cancer Re.
`search, Cell Growth & Differentiation, Cancer Epidemiology, Biomarkers & Prevention, and Clinical Cancer Research
`and in nonmember subscriptions to Cancer Res
`earch. Volume 38 ofthe Proceedings of the American Association for Cancer Research (ISSN 0197-016X) succeeds
`Volume 37 of the Proceedingsof the American
`Association for Cancer Research. The Proceedings may be obtained at a price of $45.00 throughregistration at the
`annual meeting of the American Association fo
`t Cancer Research, April 12-16, 1997, or ordered by writing to: Proceedings of the American Association for Cancer
`Research, P.O. Box 3000, Denville, NJ 07834
`(Telephone: (800) 875-2997 or (201) 627-2427; FAX: (201) 627-5872]. Add $6.00 for shipping for orders from outside
`the U.S.; expedited delivery rates are available
`upon request.
`TheProceedings of the American Association for Cancer Researchis c
`Proceedingsorof any materials in the Proceedings
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`ay be granted contingent upon paymentof an appropriate fee. Reproduction requests mustinclude a brief description of intended
`approval of the authors before corresponding
`unauthorized duplication of anyportion of
`with the AACR.Failure to comply with the foregoing restrictions and
`these materials are a violation of applicable laws and m
`ay be subjectto criminal prosecution andcivil penalties.
`No responsibility is accepted by the Editors, by the American Association for Cancer Research,Inc., or by CadmusJournal Services for the opinions
`expressed by the contributors or for the contents of the advertisements herein.
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`ii
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`This material maybe protected by Copyrightlaw (Title 17 U.S. Code)
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`
`PHARMACOLOGY/THERAPEUTICS (PRECLINIGAL AND CLINICAL) 44
`
`(cp31 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 carcinomain the lung.
`
`#4246 CGP 57148B,a protein-tyrosine kinase inhibitor with potential for
`the treatment of Bcr-Abl positive leukemias and diseasesinvolving deregu-
`lation of PDGFreceptor and c-Kit tyrosine kinases. Buchdunger, E., Zimmer-
`mann, J., Mett, H., Muller, M., Law, N., Cioffi, C., Druker, B., and Lydon, N. Ciba
`Pharmaceuticals Division, Oncology Research Department, Ciba-Geigy Ltd., CH-
`4002 Basel, Switzerland, Ciba Pharmaceuticals Division, Ciba-Geigy Ltd., Sum-
`mit, New Jersey 07901, Oregon Health Sciences University, Portland, Oregon
`01
`ice 57148B hasbeenidentified as a potent protein-tyrosine kinase inhibitor
`with selectivity for the Abl and PDGFreceptortyrosine kinases. Cellular prolifer-
`ation and tumor growth of Ber-Abl or v-Abl expressing cells were specifically
`inhibited by this compound. CGP 57148Bselectively inhibited PDGF-mediated
`cellular events such as PDGFreceptor autophosphorylation, inositol phosphate
`formation and c-fos induction. The compound wasa potentinhibitor of PDGF-
`mediated growth of v-sis transformed BALB/c 3T3 cells and PDGFstimulated
`proliferation of A10 rat aortic smooth muscle cells. /n vivo, CGP 57148Binhibited
`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 TypeIll
`receptor tyrosine kinases, CGP 57148B was found to potently inhibit the SCF
`receptor, c-Kit, but not
`the related c-Fms, Flit-3 and Fit-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 NCI-H69 and NCI-H209 SCLClines wassensitive
`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 transductioninhibitor which might have therapeutic potentialfor the treat-
`ment of Ph+ leukemias as well as diseases which involve abnormal activation of
`PDGFreceptor and c-Kit tyrosine kinases.
`
`#4247 Cytoxicity and phosphotyrosine effects of c-src kinase inhibition
`by substituted pyridopyrimidine tyrosine kinase inhibitors in human colon
`carcinomacell 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, S., and Hamby, J. Parke-
`Davis Pharm Res. Div. of Warner-Lambert Co., Ann Arbor, MI 48105
`Tyrosine kinaseinhibition has beenidentified as a potential therapeutic strategy
`in neoplastic disease. Potent c-src kinaseinhibitors (IC;7=9nM in isolated c-src
`assay for 6-(2,6-dichloro-phenyl)-2-[4-(2-diethylamino-ethoxy)-phenylamino]-8H-
`pyrido[2,3-d]pyrimidin-7-one dihydrochloride)(PD166285, A) are described. Src
`family members fyn,
`lyn, and Ick are also potently inhibited in vitro. A causes
`growth delay on plastic with IC;, 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 ICs, values of 1.8 pM(HT-29),
`780nM(HCT-8), and 330nM(SW-620). Phosphotyrosine(P-Y) content of focal ad-
`hesion kinase(FAK) and paxillin(known substrates of c-src) is reduced at sub-uM
`concentrations in HT-29 cells treated with A for 2 hours and then mitogenically
`stimulated. In a fibroblastcell line overexpressing epidermal growth factor recep-
`tor(EGFr) and c-src, A is a specific c-sre 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 tumorcells. Iwata, K., Miller,
`P.E., Barbacci, E.G., Arnold, L., Doty, J., DiOrio, C.1., 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-358,774inhibits isolated human EGFR TK with an ICzo of 2 nM, and reduces
`EGFR autophosphorylation in MDA-MB-468 tumorcells in culture with an ICs, of
`20 nM. EGFR TK is >1000-fold moresensitive to CP-358,774 than other TKs such
`as humanc-sre, insulin receptor, insulin-like growth factor receptor,or v-abl. As a
`further indication of selectivity, EGF-induced tyrosine phosphorylation of SHC
`Proteins in HN5cells 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 ICs, of 70 nM, but doesnot
`Similarly inhibit mitogenesis stimulated by platelet derived growth factor, insulin-
`like 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-
`Pletely inhibits EGF-induced autophosphorylation of human EGFR in HN5 tumor
`xenografls and of EGFR in mouseliver. This inhibitor has potential for the
`treatment of tumors that are dependent on the EGFR pathwayforproliferation.
`
`#4249 Therapy of human carcinomasin athymic micebyinhibition 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 Iwata, K. Dept. of Cancer, Pfizer Central Research,
`Groton, CT 06340, Oncogene Science, Inc., Uniondale, NY 11553
`Tyrosine phosphorylation of EGF receptors is an importantearly event in signal
`transduction and tumorcell 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),
`weobserved reproducible ED;,’s and haveusedthis assay to describethe extent
`and duration of drug action in vivo. CP-358774 is an effective, orally active
`inhibitor of EGFr tyrosine phosphorylation (ED;)= 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 degreeofinhibition of EGFr-
`dependent human HN5 tumor growth in a xenograft therapy model (ED;,= 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
`EGFreceptor: A binding model and structure-activity relationships for sol-
`uble analogues. Denny, W.A., Palmer, B.D., Rewcastle, 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-Kallmeyer, 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-Anilinopyrido[d]pyrimidines are potent, selective inhibitors of the tyrosine
`kinase activity of the epidermal growth factor receptor (EGFR), binding at the ATP
`site with IC, values downto 8 pM. Theyare ofinterest as anticancer drugs, since
`EGFR is over-expressed in many cancers and associated with poorclinical
`prognosis. A model for their binding to the EGFR was constructed from the
`catalytic subunit of the related cAMP-dependentprotein kinase, and from struc-
`ture-activity data. This attributes their high selectivity to binding of the 4-anilino
`ring in a hydrophobic pocket of composition unique to the EGFR. The 6- and
`7-positions of the pyrido[d]pyrimidines occupy the entrance of the ATP binding
`pocket, with the 7-position pointing towards the ribose binding site of ATP. In
`agreementwith 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-phosphorylation 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 factortyrosine 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 EGFRtyrosine kinasein vitro and of the EGF-stimulated
`growth of KB oral carcinomacells in culture (0.08 micromolar). ZD1839 has
`excellent oral bioavailability and shows antitumor activity in a broad range of
`humansolid tumor xenografts implanted in nude mice in the dose range 12.5 -
`200 mg/kg once per dayorally. Sensitive tumors include A431 vulval, KB oral,
`A549 NSCLC, DU145 prostate, HT29, HCT15, CR10 and LoVocolorectal, 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-clinical data ZD1839 has been
`selected for clinical development.
`
`#4252 Novel benzoylacetylenic compounds: Potent and selective EGF
`receptor tyrosine kinaseinhibitors. Suzuki, T., Kitano, Y., Ohya, J., Umeki, H.,
`Inokawa, 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-
`mentof cancer. We have synthesized a novel series of benzoyl acetylenic com-
`poundsaspotent and selective inhibitors of EGF receptor tyrosine kinase. We
`found that one of these compounds, DAB-720, has excellent biological activities
`in in vitro andin vivo studies. We show herethe 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, Ick and PKC, DAB-720
`displayed selectiveinhibitory activity against EGF receptor with IC5, of 0.070 uM
`andlittle or no inhibition for the other kinases at 50 4M, except Ick for which
`DAB-720 had an ICsp of 16 4M. Autophosphorylation of EGF receptor in A431
`cells and of HER2/Erb B2 in BT474 cells werealso inhibited over 50% at 5 wM and
`10 «M respectively. The ICs9 for KB cell growth was 0.76 ,M. 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 e Volume 38 © March 1997
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