Poster Session Topoisomerase I inhibitors Friday 1 October 155

8 and 15, every 28 days, were administered to patients (pts) with advanced 508 POSTER
solid malignancies who had received minimal prior myelotoxic therapy. A National Comprehensive Cancer Network phase II study of
Results: To date, 31 pts (median age 57, [30-75]; 17 male/14 female; gemcitabine and irinotecan in metastatic breast cancer: can
pancreas/biliary tract [15/5], colon [3], esophagus [2], NSCLC/SCLC [2/1], topoisomerase I localization predict response to irinotecan?
other [3]) have received 137 courses (range 1-22) at rubitecan dose levels
of 1.0 mg/m~/day (13 pts), 1.25 mg/m~/day (8 pts), and 1.5 mg/m~/day S. Moulder, N. Valkov, S. Minton, P. Munster, R. Lush, M. Lacevic, I. Sims,
A. Cammarata, J. Horton, D. Sullivan. H. Lee Moffitt Cancer Center and
(10 pts) with full doses of gemcitabine. First cycle DLTs have been
Research Institut, Department of Interdisciplinary Oncology, Tampa, USA
uncomplicated gr 4 neutropenia >5 days (1 pt) at 1.0 mg/m~/day; gr
3 vomiting (1 pt), gr 3/4 thrombocytopenia (2 pts) at 1.25 mg/m~/day; Background: Gemcitabine, a nucleoside analogue, and irinotecan, a
and febrile neutropenia (lpt), gr 3 transaminase elevation (1 pt) at topoisomerase I (tope I) inhibitor, have both demonstrated efficacy as
1.5 mg/m2/day. Other toxicities are mostly mild to moderate, and also single agents in patients with metastatic breast cancer and preclinical data
include non dose-limiting gr 4 neutropenia (6 pts), gr 4 thrombocytopenia indicate that the incorporation of gemcitabine into DNA enhances cleavage
(2 pt), gr 3 transaminase elevations (5 pts), gr 3 diarrhea (2 pts), and gr complexes in vitro when combined with a tope I inhibitor. Since tope I
3 vomiting, fatigue, cystitis, weight loss, and epistaxis (each 1 pt). Patient requires nuclear localization to exert its activity, predominate localization of
accrual continues at 1.0 mg/m ~ of rubitecan, which is the recommended tope I within the cytoplasm may predict for drug resistance.
phase II dose, in combination with 1000 mg/m ~ of gemcitabine. 9-NC Methods: After obtaining informed consent, 16 patients received therapy
and gemcitabine AUCs (n=23 and 14 pts, respectively) increased with with gemcitabine at 1000 mg/m ~ and irinotecan at 100 mg/m ~ on days
increasing dose levels. No drug-drug interactions were identified. Overall 1 and 8 of a 21 day cycle. Tumors from 5 patients were biopsied by
clearance of 9-NC and Gemcitabine were 7555.86±12901.74 mL/hr fine needle aspiration (FNA) prior to initiation of therapy. 2×105 cells
and 314.3±1133.5 mL/hr, respectively. Other PK parameters (9-NC and were used to create cytospin slides for immunofluorescence staining
Gemcitabine, respectively) were: T1/~, 12.9±6.5 h and 10.2±13.6 h; AUC, of tope I. A monoclonal antibody against histone was used to identify
800.5±635.9 h*ng/mL and 254,779±227,986 h*ng/mL; and Vd, 92.3±97.6 nuclei and function as an internal control for sample variation. Tope I
L and 1071.3±3247.3 mL. A partial response has been observed in 4 of was detected using the C-21 murine monoclonal IgM antibody directed
16 pts with evaluable pancreas/biliary tract cancer (25%; 95%CI, 7.3% against an epitope in the C-terminal 67 kDa. Immunofluorescence
to 52.4%), of whom 2 were gemcitabine-refractory, and also in 1 pt with was observed with a Leitz Orthoplan 2 microscope and images were
esophagus cancer. Additionally, 11 of 24 pts evaluable for tumor response captured by a CCD-camera with Smart Capture program. Quantification
have shown stable disease lasting 3+-22+ months (pancreas, 4 pts; biliary of tope I was performed on 50 randomly selected tumor cells/sample
tract, 4 pts; colon, lung, and H&N, 1 pt each). Remarkably, 12 of 16 patients with measurements confirmed by Adobe Photoshop 7.0. Each cellular
with evaluable pancreatic or biliary tumors had a partial response or durable compartment was quantified separately in pixels and nuclear/cytoplasmic
stable disease as best response. ratios were calculated individually for each cell with the mean value
Conclusions: Disease-directed evaluations of this safe and feasible for each variable listed in the table below. The ratios were plotted on
regimen are planned in pancreatic and biliary tumors, where impressive scattergrams and the mean values and standard deviations were calculated
preliminary activity has been observed. with GraphPad 4.0 software.
Results: Of the 16 patients enrolled, 14 have been evaluated for response
507 POSTER with an overall response rate of 36% (CR=0, PR=5, SD=3, PD=6). The
Role of topoisomerase I inhibition in the cytotoxic action of synthetic results of the five patients who had tissue biopsies to assess for tope I are
derivatives of the anticancer marine alkaloid lamellarin D listed in the table.
Conclusion: Preliminary results indicate that gemcitabine and irinotecan
A. Lansiaux 1, W. Laine 1, C. Tardy 1, M. Iwao~, F. Ishibashi ~, C. Bailly 1. is an active combination for metastatic breast cancer and that tope I
11NSERM U-524- Centre Oscar Lambret, Pharmacology, Lille, France; localization can be measured in breast cancer patients using immunoflu-
2Faculty of Engineering and Faculty of Fisheries, Nagasaki University, orescence in tumor samples obtained by FNA. In this limited data set,
Nagasaki, Japan the tumor sample with the highest nuclear/cytoplasmic ratio of tope I was
associated with a partial response while the lowest ratio was associated
We have recently identified the marine alkaloid Lamellarin D (Lam-D) with progression of disease.
as a novel potent inhibitor of human topoisomerase I with an efficacy
comparable to that of the reference drug camptothecin (Cancer Res. Patient no. Pixeldensity Nuclear/cytoplasmicratio* Clinical response
2003, 63, 7392-7399). This natural product is highly cytotoxic and
insensitive to P glycoprotein-mediated drug efflux; its cytotoxicity is Nuclear Cytoplasmic
Topo I Topo I
dependent, at least in part to its capacity to promote DNA cleavage by
topoisomerase I. In the present work, we have analyzed the topoisomerase
003 76646 58633 1.5 PD
I inhibitory properties of 8 lamellarin derivatives diversely substituted on
the benzopyranopyrroloisoquinolinone B-F pentacyclic planar chromophore 004 74707 90233 0.91 PR
006 150114 149439 1.23 SD
or the orthogonal phenol A-ring (J. Nat. Prod. 2002, 65, 500-504).
010 53411 6531 13.5 PR
Stabilization of topoisomerase I-DNA covalent complexes was studied
013 79158 184587 0.5 PD
using complementary electrophoretic methods with supercoiled plasmid
and radiolabeled DNA restriction fragments. The cytotoxicity of the test
compounds was evaluated by a conventional tetrazolium-based assay
using a pair of cell lines expressing a normal or mutated topoisomerase I 509 POSTER
gene. Human CEM leukemia cells are highly sensitive to Lam-D whereas Antitumour activity of the novel 7-substituted camptothecin ST1481
the CEM/C2 cells resistant to camptothecin are cross-resistant to Lam-D. (Gimatecan) in human neuroblastoma
The mutation of the Asn722 to a Ser residue adjacent to the active
site Tyr723 residue of the human topoisomerase I enzyme considerably A.M. Di Francesco 1 D. Meco 1, A. Riccardi 1, G. Barone 1, C. Pisano z,
decreases the cytotoxicity of Lam-D and its analog FI-02 lacking a methoxy P. Carminati z, S. Rutella 3, M. D'lncalci 4, R. Riccardi 1. 1Catholic University,
group on the F-ring. In contrast the deletion of the adjacent hydroxy Division of Pediatric Oncology, Rome, Italy; 2Sigma- Tau, Dept. of Oncology
group considerably reduces the cytotoxicity of the compound and almost R&D, Pomezia (RM), Italy; 3Catholic University, Division of Hematology,
abolishes its ability to interfere with topoisomerase I. The hydroxyl group
Rome, Italy; 4Institute "M. Negri", Dept. of Oncology, Milan, Italy
on the phenol A ring is also a crucial element both for cytotoxicity and
topoisomerase I inhibition. This study (i) reveals a solid correlation between
the cytotoxic potential of the 8 lamellarin derivatives tested and their ability Background: Gimatecan (ST1481,7-tert-Butoxyiminomethylcamptothecin),
to inhibit topoisomerase I, and (ii) provides important structure-activity is a novel lipophilic camptothecin analog showing a better pharmacological
relationships to guide the development of antitumor agents in this chemical profile and a lack of cross-resistance to topotecan and irinotecan.
series. Gimatecan is currently under evaluation in Phase 1/11 clinical trials
administered by oral route. In the present study we compared the in vitro
antitumour activity of gimatecan, SN38 (the active metabolite of irinotecan)
and topotecan in neuroblastoma.
Methods: Cytotoxicity was evaluated by growth inhibition assay and
clonogenic survival in a panel on neuroblastoma cell lines (SK-N-DZ;
BE(2)M17; LAN-1; RNGA and BE(2)c). From these studies SK-N-DZ
cells were selected for further evaluation of cell cycle distribution by flow
cytometry; induction of DNA strand-breaks induction by alkaline Comet
assay; induction of apoptosis through the hypoploid peak, active caspase-3