DC beads loaded with Irinotecan : precilinical in-vitro & in-vivo evaluation

1. DC beads loaded with Irinotecan :precilinical in-vitro &in-vivo evaluation T de Baere • Bench top • Animal model

2. Colorectal cancer chemotherapy % response rate FOLFIRI FOLFOX LV5FU2 5FU-FA 55 % 55 % 5 FU 25 % 20 % 10 % Sytemic therapy • Improvement in response rate, namely due to Oxaliplatinum and Irinotecan

3. Colorectal cancer chemotherapy % response rate Oxal. FOLFIRI FOLFOX FUDR 64 % LV5FU2 5FU-FA 55 % 55 % 45 % 5 FU 25 % 20 % 10 % Sytemic therapy Intra-arterial therapy • Intra-arterial delivery provides higher response rates • 50% of the responder to IV oxaliplatin had failed IV oxalipaltin and irinoteacn (Boige V, Lacombe S, de Baere T - Ann SurgOncol 2008)

4. CL Rart = QA (1 - Er) Rationale for intra-arterial route - Rart : advantage of IA vs IV -Er :extraction ratio at 1st pass - QA : arterial flow - CL : Body clearance of the drug

5. CL Rart = QA(1 - Er) 101 81 Rart 61 0.1 41 Er 0.5 21 0.9 1 24 18 12 6 QA (L/h) Rationale for intra-arterial route • Drug eluting embols • Complete embolization (QA=0, Rart≈∞)

6. In vitro (loading) Loading capabilities of embolics with Irinotecan Irinotecan - 25mg/ml of beads •  DC beads™ • 500-700 mm hydrated •  Hepaspheres™ • 100-150 mm dry • 400-600 mm hydrated Comparative study of chemoembolization loadable beads. Jordan O, Denys A, de Baere T, Boulens N, Doelker E. JVIR. 2010

7. In vitro (Rigidity) Loading capabilities of embolics with Irinotecan Irinotecan - 25mg/ml of beads •  DC beads™ • 500-700 mm hydrated •  Hepaspheres™ • 100-150 mm dry • 400-600 mm hydrated Compression of a microsphere monolayer to determine elastic modulus Comparative study of chemoembolization loadable beads. Jordan O, Denys A, de Baere T, Boulens N, Doelker E. JVIR. 2010

8. In vitro (Size) Loading capabilities of embolics with Irinotecan Irinotecan - 25mg/ml of beads •  DC beads™ • 500-700 mm hydrated •  Hepaspheres™ • 100-150 mm dry • 400-600 mm hydrated • Shrinkage under irinotecan loading with return to baseline after release Comparative study of chemoembolization loadable beads. Jordan O, Denys A, de Baere T, Boulens N, Doelker E. JVIR. 2010

9. In vitro (loading homogeneity) Loading capabilities of embolics with Irinotecan Irinotecan - 25mg/ml of beads •  DC beads™ • 500-700 mm hydrated •  Hepaspheres™ • 100-150 mm dry • 400-600 mm hydrated Comparative study of chemoembolization loadable beads. Jordan O, Denys A, de Baere T, Boulens N, Doelker E. JVIR. 2010

10. In vitro (Elution/Release)  Hepaspheres  DC beads DC Beads Hepasheres Drug load = 93% Drug release = 102 ±11 % T75% = 66 minutes Drug load = 90% Drug release = 95 ± 11 % T75% = 7 minutes Comparative study of chemoembolization loadable beads. Jordan O, Denys A, de Baere T, Boulens N, Doelker E. JVIR. 2010

11. In vivo • VX2 liver tumor • IV : 12 mg • IA : 12 mg • TACE : 6-16.5 mg (m=9.35) / 100-300 microns • Complete stasis • PharmacoKinetic CPT11 & SN38* • Venous sampling (10min - 24H) • Tissue sampling (1, 6, 24H) • Tumor • Liver • Quantification of tumor necrosis * SN38 is the most active metabolite of Irinotecan

12. In vivo : venous concentration Serumirinotecan IV IA DEB • Reduced systemic peak levels by more than 50% • Lower systemic toxicity expected for a given dose

13. In vivo : venous concentration Serumirinotecan Taylor RR, Eur J Pharmaco Sciences 2007 • Reduced systemic peak levels by more than 50% • Lower systemic toxicity expected for a given dose

14. In vivo : venous concentration Serumirinotecan • Complex SN38 pattern, but still lower systemic level for DEB • SN38 : active metabolite through carboxylesterase in the liver

15. In vivo : Tissue concentration IV IA DEB

16. In vivo : Tissue concentration • Higher concentration of Irinotecan in tumor at 24 hours • DEBIRI : X 10 IA • DEBIRI : X 64 IV

17. In vivo : Tissue concentration • Higher concentration of SN38 at 24 hours

18. In vivo : Tissue concentration CRC rat model  48h Irinotecan  72h Irinotecan Concentration (nM) MTT assay (in vitro) shows 50% cell survival with 50 g at 48 h, and 25 g at 72 h in CC531-lac-Z rat CRC cells Eyol E, Clin Exp Metastasi 2008.

19. IV Tumor necrosis IA Tumor necrosis DEB • Baseline necrosis of 30%, usual in VX2 tumor • Equivalent percent of necrosis for IA & TACE @ 6 h • Significantly higher necrosis for TACE @ 24 h

20. Liver toxicity • ≈ 10 fold increase in transaminases • Start to decrease as early as 24 hours

21. Conclusion • In vitro • DC Beads can load rapidly 96% of Irinotecan • DC Beads provide a real delivery platform for Irinotecan without burst release • In vivo • DEBIRI produces lower systemic exposure to Irinotecan when compare to IV or IAH injection • DEBIRI provides higher Irinotecan and SN38 concentration in tumor than IV and IAH • DEBIRI provides higher degree of tumor necrosis than IV or IAH injection of Irinotecan

23. In vitro (Elution/Release) Flow • flow-through apparatus 4 (Sotax CE6; Sotax) • Six parallel implant cells. Flow at 5 mL/min • Faster release under flow than under simple shaking Comparative study of chemoembolization loadable beads. Jordan O, Denys A, de Baere T, Boulens N, Doelker E. JVIR. 2010

24. Colorectal cancer • World wide incidence: 400,000 patients • 60% patients will develop liver metastases • Surgery is feasible in only a minority of patients and most patients are treated with systemic chemotherapy. • Patients have a poor prognosis with reported 1- and 3-year survival rates of 31% and 2.6%, respectively .

25. In vivo : Tissue concentration Activation of carboxylesterase by embolization ?