Viability of Microorganisms in Novel Chemical and Biopharmaceutical Anticancer Drug Solutions
Total Page：16
File Type：pdf, Size：1020Kb
European Journal of Parenteral & Pharmaceutical Sciences 2015; 20(1): 5-12 2015 Pharmaceutical and Healthcare Sciences Society Viability of microorganisms in novel chemical and biopharmaceutical anticancer drug solutions Iman Sarakbi, Matteo Federici, Irene Krämer* Department of Pharmacy, University Medical Center Mainz, Johannes Gutenberg-University, Mainz, Germany Most anticancer drug products used in clinical practice lack antimicrobial properties. Therefore, materials and methods utilised to prepare the parenterally administered preparations must ensure sterility and avoid the introduction of contaminants and the growth of microorganisms. The aim of the study was to evaluate the growth potential of four different microorganisms in diluted ready-to- use novel chemical and biopharmaceutical anticancer drug preparations. In three consecutive series, 14 different antineoplastic drugs were diluted to the lowest customary concentrations in polyolefin containers prefilled with 0.9% sodium chloride or 5% dextrose solution. Aliquots (9 mL) of each anticancer drug solution were inoculated with 1 mL suspension of bacteria or fungi (Staphylococcus aureus, Enterococcus faecium, Pseudomonas aeruginosa and Candida albicans) to achieve approximately 104 microorganisms per mL. Pure vehicle solutions were used as positive controls in each series. The inoculated preparations were stored at room temperature (22°C) and protected from light. Samples (1 mL) were taken immediately and 1, 3, 5, 24, 48 and 144 hours after inoculation, processed and transferred to tryptic soy agar plates. The plates were incubated at 37°C and the colony-forming units counted after 24 hours. The tested microorganisms remained viable in most of the anticancer drug solutions over a period of 144 hours after inoculation. Trabectedin was the only product generating distinct and rapid antibacterial activity. Viability of C.albicans was not affected by trabectedin, but growth of the fungus was retarded in temsirolimus-containing samples. Nab-paclitaxel suspension supported the growth of the selected bacteria and fungus. Most of the novel anticancer drug products showed neither growth-retarding nor growth- supporting properties. Therefore, in pharmacy departments the anticancer drug products for parenteral administration should be prepared under strict aseptic conditions and refrigerated. Lack of antibacterial and antifungal properties should be considered when assigning extended expiry dates. Attention should be paid to the vulnerability of albumin-containing nab-paclitaxel suspensions to microorganism proliferation. Key words: Biopharmaceutical anticancer drug solution, microorganisms, antimicrobial activity. 1 Introduction 010-3 , the European Pharmacopoeia 7.7 Pharmaceutical Preparations2, the United States Pharmacopeia Monograph The armamentarium of anticancer drugs is still constantly <797>3, and various national guidelines. Materials and growing. Recently approved anticancer medicinal products methods used to prepare parenterally administered products comprise of the innovative signal transduction inhibitors, must ensure sterility and avoid introduction of contaminants which can be administered orally, as well as novel small and the growth of microorganisms2. molecules and monoclonal antibodies, which are administered Quality tests of individual anticancer drug preparations are parenterally. The latter are mostly supplied as ready-to-use limited by singularity of each preparation and time restraints. preparations individually prepared for specific patients by In most cases, preparations are administered earlier than the pharmacy-based centralised cytotoxic preparation units. The results of sterility tests are available. However, the results of preparation process must take place in accordance with the sterility tests, media fills and environmental monitoring regulatory framework of good preparation practice, such as the programs are useful for process validation. Pharmaceutical Inspection Cooperation Scheme (PIC/S) PE This is especially true because most of the anticancer drug preparations lack antimicrobial properties4–7. Of note, growth *Corresponding author: Professor Dr. Irene Krämer, Department of of microorganisms was not stimulated in previously tested Pharmacy, University Medical Center, Johannes Gutenberg-University, 4 Langenbeckstraße 1, 55131 Mainz, Germany; Tel: +49 6131 177209; Email: monoclonal antibody-containing preparations . There was no [email protected] obvious direct link between chemical structures or 5 6 IMAN SARAKBI, MATTEO FEDERICI, IRENE KRÄMER pharmacological principles and antimicrobial activity in Pharmacy Department of the University Medical Center our previous studies. Moreover, there was no correlation Mainz, Germany. Vehicle solutions and container material between antifungal and antibacterial activity, and were selected in order to guarantee maximal chemical and antimicrobial activity proved to be species specific. physical stability. Samples were prepared by injecting the During the validation of the sterility tests, the calculated amount of each concentrated solution into a manufacturers of medicinal products have to determine polyolefin bag containing a nominal volume of 50 mL the extent to which bacteria survive in the products. The 0.9% NaCl infusion solution (Freeflex Isotonische aim of the present study was to evaluate the growth of four Kochsalzlösung Infusionslösung, Fresenius Kabi different microorganisms in diluted ready-to-use Deutschland GmbH, Bad Homburg, Germany; LOT preparations of 14 chemical and biopharmaceutical 13GBS271, LOT 13GGS152) or a polyolefin bag medicinal products approved for anticancer therapy after containing a nominal volume of 250 mL 5% the year 2007. The microorganisms chosen are commonly dextrose/glucose solution (Freeflex Glucosteril® 5% associated with nosocomial infections and represent Infusionslösung, Fresenius Kabi Deutschland GmbH, Bad potential contaminants. The test conditions simulate the Homburg, Germany; LOT 14GD7107) in order to achieve worst possible conditions for patients and optimal the predetermined concentrations. These final circumstances for the growth of microorganisms. concentrations were the lowest ones prescribed by the physicians in daily clinical practice. ® Materials and methods Nelarabine injection solution (Atriance ) was transferred to an empty infusion bag (MIB300 - EVA The study was performed in three consecutive series, mixing and infusion bag, Hegewald Medizinprodukte investigating the antimicrobial activity of four drug GmbH, 09638 Lichtenberg, Germany; LOT 3GH71302), products in the first series (aflibercept, brentuximab- and used without any further dilution. Nab-paclitaxel vedotin, nelarabine, ofatumumab), four drug products in (Abraxane®, Celgene, München, Germany) suspension the second series (arsenic trioxide, temsirolimus, was prepared by adding 20 mL of 0.9% NaCl infusion ipilimumab, cabazitaxel), and six drug products in the solution (Freeflex Isotonische Kochsalzlösung third series (trabectedin, eribulin, fotemustine, nab- Infusionslösung, Fresenius Kabi Deutschland GmbH, Bad paclitaxel, panitumumab, clofarabine). During each of the Homburg, Germany; LOT 14GG7346) to 3 vials each and three series, the vehicle solutions (0.9% sodium chloride 7 mL aliquots were withdrawn directly from the vials. (NaCl) solution, 5% dextrose solution) were tested in Aliquots amounting to 9 mL were aseptically parallel and acknowledged as a positive control measure. withdrawn in quadruplicate from each sample preparation (except nab-paclitaxel) and transferred to sterile empty Preparation of inocula containers (15 mL centrifuge tubes with screw caps, VWR The preparation of inocula was carried out at the Institute International, Radnor PA, USA; LOT 317CE). of Medical Microbiology and Hygiene, University Medical Center Mainz, Germany. The four Inoculation microorganisms selected for the study were Prior to inoculation, each tube containing bacterial or Staphylococcus aureus ATCC strain 6538, Pseudomonas fungal suspension was carefully homogenised with an aeruginosa ATCC strain 15542, Enterococcus faecium automatic shaker for 2 minutes in order to suspend CFU ATCC strain 6057, and Candida albicans ATCC strain adhering to surfaces and to ensure the uniformity of the 10231. The microorganisms were cultured on agar plates suspensions. Subsequently, 1 mL of the suspension was (CASO-Agar, Oxoid Deutschland GmbH; LOT 1406767) added to 9 mL of the freshly prepared drug product to for 24 hours (C. albicans 72 h) at 37°C. achieve concentrations of approximately 104 Subsequently, the cultures were harvested and microorganisms per mL. As an exception, 7.0 mL of nab- suspended in 0.9% NaCl solution. The concentration of paclitaxel suspension was inoculated with 0.7 mL of microorganisms was adjusted by matching the turbidity of bacteria or fungal suspension to achieve the target the suspensions with McFarland standards. The concentration of microorganisms. McFarland standards used to achieve a microorganism NaCl solution (0.9%) and 5% dextrose solution (same 8 concentration of 10 colony forming units (CFU)/mL were brand and batch number as the vehicle solution) 0.5 for S.aureus and E.faecium, 0.2 for P.aeruginosa, and inoculated with microorganisms in the same manner, 2.5 for C.albicans. The inocula were further diluted with served as positive controls. Dextrose solution (5%) served 0.9% NaCl solution to achieve viable counts of as a positive control for the fotemustine