Short-term storage stability of Triple IT solution Triple IT solution Objective Stability protocol Results Discussion Conclusion Matthias D’Hondt1, Elien Vangheluwe1, Nadia Lemeire1, Tiene Bauters2, Brigitte Pelfrene2, Johan Vandenbroucke2, Hugo Robays2, and Bart De Spiegeleer1,* 1 DruQuaR (Drug Quality and Registration) group, Faculty of Pharmaceutical Sciences and 2 Ghent University Hospital, Ghent University, Harelbekestraat 72, B-9000 Ghent, Belgium (*Bart.DeSpiegeleer@UGent.be) 2011-211b

Triple Intrathecal (IT) chemotherapy: Triple IT solution Objective Stability protocol Results Discussion Conclusion Triple Intrathecal (IT) chemotherapy: Cytarabine (CB): pyrimidine antitumor agent Methotrexate (MT): synthetic inhibitor of dihydrofolate reductase vomiting, fever, paraplegia, meningoencephalopathy corticosteroid: reduces complications BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Hydrocortisone: benzylalcohol (preservative) Triple IT solution Objective Stability protocol Results Discussion Conclusion Hydrocortisone: benzylalcohol (preservative) Methylprednisolone (21)-sodium succinate (Solu-Medrol®): no preservative Yet, no in-use stability data available: logistics and cost issues BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

(0.385 %m/V CB, 0.154 %m/V MT, 0.051 %m/V MP(21)-SS) Triple IT solution Objective Stability protocol Results Discussion Conclusion Determination of short-term (in-use) storage stability of Triple IT solution (ex tempore prepared): (0.385 %m/V CB, 0.154 %m/V MT, 0.051 %m/V MP(21)-SS) Evaluation of influences: - Storage temperature - Photometric conditions - Packaging - Batch to batch variability BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

1. Syringe system (PhaSeal®) 2. Brown glass vial Triple IT solution Objective Stability protocol Results Discussion Conclusion Batch 3 different batches Packaging material 1. Syringe system (PhaSeal®) 2. Brown glass vial 3. Brown glass vial filled with needles BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Temperature (°C) Relative humidity (%) Triple IT solution Objective Stability protocol Results Discussion Conclusion Temperature stability time points (n=6): 0, 4, 8, 24, 32, 48 hrs. 3 batches, 3 packaging materials (all protected from light) Temperature (°C) Relative humidity (%) 5 50 25 60 40 75 Controlled conditions BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Photometric stability Triple IT solution Objective Stability protocol Results Discussion Conclusion Photometric stability - Triple IT solution  quartz cuvette (25°C / 60 % R.H.) UV light – 2 days VIS light – 8 days - (Light protected) control samples 2 batches, in duplo ICH Q1B compliant (light energy) BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Fused-core HPLC method Triple IT solution Objective Stability protocol Results Discussion Conclusion Fused-core HPLC method Column (+ guard column) Halo C18 (4.6×150 mm; 2.7 µm) Column / sample temp. 30°C / 15°C Mobile phase A: 0.1% V/V glacial acetic acid in H2O B: 0.1% V/V glacial acetic acid in ACN Gradient program Time (min) 15 %A 90 10 %B Flow rate 1 ml/min Injection volume 10 µl Detection DAD UV @ 240 nm (MP(21)-SS) @ 280 nm (MT and CB) BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University Triple IT solution Objective Stability protocol Results Discussion Conclusion T0hrs. Triple IT 240 nm 280 nm BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Triple IT Comp. Degradation Impurity formed Mass balance 1 Triple IT solution Objective Stability protocol Results Discussion Conclusion Temperature stability – Global results Triple IT Comp. Degradation Impurity formed Mass balance 1 CB - 100.39 – 101.31% MT 99.43 – 100.97% MP(21)-SS 2 stat. significant RRT: 0.88 and 0.90 99.49 – 100.40% 2 Reporting threshold: 0.5% 1 relative to T0 hrs.; 95% CI 2 Including formed impurities, assuming RRF 1 BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

MP(21)-SS degradation kinetics: Triple IT solution Objective Stability protocol Results Discussion Conclusion MP(21)-SS degradation kinetics: assuming (and confirmed) 1st order to determine k Arrhenius regression: ln k = ln A – Ea/RT   Ea: 45.5 kJ/mol (95% CI: 41.6 - 49.4 kJ/mol) A: 524×103 h-1 (95% CI: 107×103 - 2571×103 h-1) BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Identification MP(21)-SS related degradants (ESI-MS) [M+H+]+ [M+H+]+ RRT: 0.88 RRT: 0.90 Methylprednisolone Methylprednisolone (17)-succinate [2M+H+]+ [2M+H+]+ BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Batch / packaging material influence on MP(21)-SS degradation Triple IT solution Objective Stability protocol Results Discussion Conclusion Batch / packaging material influence on MP(21)-SS degradation MLR: k batch and packaging material (Temp. constant)  No stat. sign. (95%) batch / packaging influence BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Triple IT Comp. Degradation (%) 1 Impurity formed Mass balance 1 Triple IT solution Objective Stability protocol Results Discussion Conclusion Photometric stability – Global results Triple IT Comp. Degradation (%) 1 Impurity formed Mass balance 1 CB UV - 101.29 VIS 3.80 RRT 1.12 99.47 MT 32.46 66.92 100.00 RRT 0.85; 1.05 28.09 MP(21)-SS 2 RRT 0.88; 0.90 98.62 90.84 Reporting threshold: 0.5% 1 Decrease of API, relative to light protected control samples 2 Including formed impurities, assuming RRF=1 BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Photometric stability – MTdegradation Triple IT solution Objective Stability protocol Results Discussion Conclusion Photometric stability – MTdegradation MT: single compound vs Triple IT mixture BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Related impurities specifications (Ph. Eur.) of MP(21)-SS MP: 0.5% Triple IT solution Objective Stability protocol Results Discussion Conclusion Related impurities specifications (Ph. Eur.) of MP(21)-SS MP: 0.5% MP(17)-SS: 0.5% Risk: MP (lower solubility) precipitation Risk: min. (31% degradation) Conclusion: some consistent and controlled (MP + MP(17)-SS) degradation of MP(21)-SS is allowed in FPP (DP)  Triple IT (3%). In API  Solu-Medrol®  FDP (Triple IT) higher, e.g. 1% ∑impurities < 1% BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Predicted MP(21)-SS degr. in function of storage condition and time Triple IT solution Objective Stability protocol Results Discussion Conclusion Predicted MP(21)-SS degr. in function of storage condition and time BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Related impurities specifications (Ph. Eur.) of MT API Triple IT solution Objective Stability protocol Results Discussion Conclusion Related impurities specifications (Ph. Eur.) of MT API 0.2% (= IT): H, I 0.3%: (B, E) 0.5% (= QT): C 3%: (F) 6 unspecified  0.05% ∑impurities < 0.5% (excl. imp. B, E and C) Conclusion: no significant MT degradation is allowed  Triple IT: total imp. <0.5% 6 specified  BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Proposed storage conditions: Storage: Triple IT solution Objective Stability protocol Results Discussion Conclusion Proposed storage conditions: Storage: Max. 12 hour refrigerator  1.8% MP(21)-SS degradation Before administration Max. 30 min heating cycle (37°C)  0.6% MP(21)-SS degradation During administration / transport Max. 2 hours administration (25°C)  1.1% MP(21)-SS degradation All light protected  no MT degradation BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University

Thank you for your attention!!! BGFW - DruQuaR, Fac. Pharm. Sciences, Ghent University