P-949 Phase II pharmacodynamic trial of erlotinib in advancednon-small cell lung cancer (NSCLC) patients previously treated with platinum-based chemotherapy

Posters I Targeted therapies Results: Based on the flow cytometnc analys~s. EPCs wore increased by 4-fold in cancer patients (P < 0 0002 vs healthy controls) Differences between patients with adeno- or squamous cell carcinoma wore not significant Before therapy, the number of EPCs did not correlate with the stage of the disease. however in the subgroup of 12 patients who achieved a complete remission. the EPC number in peripheral blood was similar to healthy contToIs and significantly lower than eancer4)eadng patients Interestingly. circulating RNA levels of endothelial progenitor call-specific markers CD133. VE-Cadherin. and CD34. were not significantly increased in NSCLC patients, whereas circulating VLGFR2 RNA was significantly increased in cancer patients before therapy (P <0.001 vs. healthy controls). Furthermore. a statistically significant increase was seen in the expression of VLGFR2. RNA in stage IV. patients relative to stage I. patients. In surgically treated patients with high preoperative clroulatlng EPC number. we analyzed NSCLC tissue samples for the frequency and disthtxJtlon of EPCs incorporated into the endothelial layer of tumor vessels CD31 and the eady EPC marker CD133 double labeling of tumor microvessels made dear that incorporated EPCs are extTemely rarely can be found in NSCLC tissue sections Conduslon,=: Our findings demonst]'ate the significant increase of late circulating EPCs. and the increased expression of the EPC specific marker 'v'EGFR2 RNA in patients with NSCLC While these data suggest a participation of endothelial progenitor cells in tumor growth and vasculartzatlon in these patients, it is not clear yet. whether EPCs are essential for these processes or what the relative conthbution of ECP is compared with that of in sltu proliferating endothelial coils. Moreover. it still remains to be determined whether EPCs can be targeted to t]'oat certain typos of malignancies, or alternatively as they are endowed with the capacity to home to lung cancer vasculature they can be used to deliver toxins or vascular targeting agents. [P~

Bexarotene and erlotlnlb as an active targeted cornblnaUon therapy for advanced aerodlgastlve 'b'act cancer

K. Dragnev I . W. PettyI . S. Shah2. M. Carey 2. W. DiSaivo <. I. Williams 2. J Riges I . E Dmitrovsky2 lOartmou~4-titchcock Medical Center, Lebanon,

New Hampshire, USA, 2Dartmouth Medica/ School, Hanover, NH, USA Background: We previously reported the abberant expression of cyclin D1 and the epidermal growth factor receptor (EGFR) as early events in lung carcinogenesis. In human bronchial epithelial cells (HBEC) all trans retinoic acid (RA) suppressed EGFR expression through a b'anscrlptional mechanism and cydin D1 through proteasome dependent proteolysis. Non classical retinolds such am the rexinold, bexaroteno (B). repress both cyclin D1 and EGFR expression but signal independent of RAR beta. which is often transcriptionally silenced in lung cancer Combining an EGFR tyrosine Idease inhibitor, edotlnib (E). with B induced at least addi'dve suppression of growth and o/clin D1 expression in lung cancer cell lines and in RA-resistant HBEC with silenced RAR beta Methods: We conducted a phase I tTial of E and B in patients (lots) with advanced aerodigestive tract cancers Primary obje~ve was to determine the maximum tolerated dose of the combination Secondary objectives were determination of texicities, effK~Cy, and surrogate markers of response (cydin D1) in bucoal swabs from lots at the highest dose. Three dose levels were studied ndash: 1: B 300m~'m 2. E 100rag: 2: B 300mg/m 2. E 150mg: 3: B 400mg/m 2. E 150mg. At least 3 pts were enrolled at each level. All pts received atorvastatin for elevated tnglycendos. Results:Twenty four pts were enrolled incluclng 46% women and 79% with NSCLC. The median age was 61 years. 62% had first line and 21% second line chemotherapy, one third had KPS of 70. and 67"% were former smokers Median time on tTeatment was 46 days Tmdcitles ware mild with frequent hyperlTiglycandemia and skin rash No cases of pancreatltls were observed Two DLTs wore observed: CPK elevation and systemic pain Four objec~ve papal responses were seen in lots with NSCLC. eight pts (,5 NSCLC. 3 head/neck) had stable cisease With a median follow-up of 8 3 months, median time to progression was 2 23 months, overall survival was 14 1 months, and on6~year survival was 73.8%. For NSCLC patients the one year survival was 72%. EGFR sequencing revealed no activating mutations in axons 18. 19. and 21 for two responding cases. Cyclin D1 protein expression was repressed in bucoal swabs from 5/'6 pts. Conclusions: The recommended phase II doses are edotlnib 150mg and bexarotene 400 mg/m 2 daily This regimen is well tolerated and shows evidence of activity for the treatment of NSCLC

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S369

Phase II pharmacobynamlc trial of erlotlnlb In advanced non-small cell lung cancer (NSCLC) patients previously treated with plaUnum-baesd chemotherapy

E Felip I . F Rojo I . A Helle~. D Fcarnzler ~. J Mejo I . B Klughammer ~. J Ramos 1. H Maacke ~. U Brennscheidt ~. J Baselga I 1Vaild'Hebron

Umverstty Hospttal, Barcelona, Spain: 2Roche GmbH, Swrlzerland, 3E HoffmannJ~a Roche Ltd, SHntzerland Background: Edctinib is a potent epidermal growth factor receptor (EGFR) tyrosin6~lUnese inilb~tor that significantly prolongs survival in 2nd and 3rd line NSCLC t]'eatment. This ongoing b-lal alms to identify preclctlve markers of clinical benefit, to study the effects of edotlnib on EGFR and downsb'eam signaling cemponents, and to estimate response rates to odotlnib in NSCLC patients who progressed after platinum based chemotherapy. Methods: All patients underwent tumor biopsy at study entry and then started Izeatmont with odotlnib 150 mg/.d p.o. In a subgroup of patients a ~rther biopsy was performed after 6 weeks of edotlnib tTeatment Results: As of December 2004. charactedstlos of the 58. patients included ware: median age ,56 (range 35-7"8): sex: male 72%. female 28.%: histology: adeneearclnoma 49%. large cell 33%. squamous cell 18% Of 49 avaluable batlents. 5 (10%) achieved partial response (PR). 1g (39%) had stable disease (SD) and 2,5 (,51%) had progression (PD) Edotlnib was well tolerated. 7"0% of patients had grade 1-3 rash and no unexpected texicities were seen On analyzing tumor samples from diagnosis. EGFR mutations were found in 2 patients with PR. No mutations were seen in 1 patient wtth SD or in 5 with PD. The effects of 6 weeks of odotlnib on EGFR pathway were analyzed in 12 patients for whom tumor samples wore available both at study entry and after 6 weeks of b-eatmont. After edotlnib t]'oatment. EGFR esprosslon remained unchanged whereas there was a decrease in phosphorylatod (p) MAPK (p= 0.002) and Ki 67 expression (p= 0.033). Additionally. in those patients with clinical benefit there was a decrease in p/~,KT expression Co 0 08.) after 6-weeks of edotinib treatment Conclusions: Edctinib achieves significant clinical benefit in praviously-tTeated NSCLC patients EGFR mutations are seen in some edotlnib responders In senal tumor biopsies, edotinib was shown to decrease levels of pMAPK and ~4~7 Accrual is ongoing to 80 patients and affymetdx analyses are to be performed [P~

Combined antlsense chemoradlolmrnunotherapy consisting of entI-HER2 scFv lifted onto high energy radioisotopes, vlnorelblne and 21-nucleoUds double stTended slRNA targeted to DNMT1 Induce PCD In metastatic NSCLC characterlsed by hypermetbylated oncosuppressor p16 promoter CpG Islands and overexpreeslon of oncogerms cdc25c, Raf-1, HER-2 and bct-2

J. Giamios ~. P. Lambrlnes 2, E. Michaila=kisI . ~Dept.ot Ct/ntcal Oncotogy,

GSHA, Athens, Greece, 2Dept of Oncology, PF, Greece Background: There is a great need for targeted and tailored treatment strategies in SCLC patients diagnosed with metastatic disease.The current conventional treatment sb'atagios consisting of surgery chomo or radiotherapy have a palliative effect leering to short survival parlods loss than two years.]he failure of tumor cells to undergo apoptosis cause resistance to chomoradologlcal therapies due to overexpresslon of antlapoptotlc oncogonos such as bcl 2. HER 2/nou(c~rbB2). odc25c or RaF1 and transcriptionally repressed antlapoptotlc tumor suppressor genes such as pl 6 due to aberrant promoter methylatlon which was associated with CIMP+ Methods: We obtain tumor calls from a metastatic NSCLC patient Methylatlon specific PCR detected methylatlon of p16 promoterQuantltatlve IHC. WB. SB and PCR exhibited overespression of DNMTI. bcl-2. HER-2Jneu(c-erbB2). Raf-1 and edc25cWe treated the NSCLC calls with antiq-lER-2(c-erbB2) scFv attached to high energy- radioisotopes, vinorelbine-tartTate and 21-nueleotlde double-stranded siRNA segment generated against DNMT1 Results: Post4reatmont. we detected r6~expression of oncesuppressor p16 after inhibition of DNMTlmRNA. downregulatlon of antlapoptotlc oncegone HER 2/nou(~orbB 2) duo to targeted scFv and inactivation of bcl 2. Ra#l and odc25c due to phosphorylatlon by vlnerelbino. Furthermore. we detected upregulatlon of p21waf1, p27kip and Bak. The high energy radioisotopes induced DNA double stz'and breaks in tumor cells arresting synergistically with MT depolymenzlng vinorelbine their growth at the G2JM Izansit]on acoording to flow oftometTy analysis We detected externalization of PS. depolarization of mitochendrial t]'ansmombrane potential (A'~M). activation of cespase3. cespase-9, bax and DNA fragmentation TEM e~d'libited irreversible D2 apoptotlc signs forming apoptotlc bodies which were phagocytosed by adjacent tumor cells leading to a bystander killing effect (BIKE) BrdlJ and MTI" eshibited ini"ibitlon of DNA synthesis and metabolic activity of tTeated tumor calls compared to untreated controls Conclusion: We have achieved to eradicate metastatic NSCLC cells with combined chomoradoimmunetherapy after c~rcumvontlon of chom~ and radores~stant mechanisms such as hypermethylatlon of oncasuppressor p16 promoter and ovormq3ression of antlapoptotlc oncogenos such am bcl 2. her 2J'neu. Ra#l and cdc25c.