¹Universitätsklinikum Jena, Institut für Krankenhauspharmazie, Jena, Germany
²Helmholtz-Zentrum für Infektionsforschung Braunschweig, Mikrobielle Interaktionen und Prozesse, Braunschweig, Germany
³Universitätsklinikum Jena, Klinik für Innere Medizin IV, Jena, Germany
⁴Universitätsklinikum Jena, Institut für Klinische Chemie und Laboratoriumsdiagnostik, Jena, Germany

Text

Background: The ongoing FRESCO trial [1] is designed to assess the efficacy and safety of long-term (12 weeks) oral administration of frozen encapsulated faecal microbiota (FM) and a sterile microbiota filtrate (FMF) compared to placebo (PL) in patients with mild to moderate active ulcerative colitis (UC).

A key challenge in the supply of frozen FM or FMF capsules is ensuring the integrity and stability of the frozen investigational medicinal product (IMP) throughout the entire supply chain – from manufacturing to administration to the patient – in order to preserve therapeutic potential and safety. After manufacture, the capsules should be stored at temperatures of ≤ -65 °C and remain frozen at ≤ -10 °C during transport and storage at the patient’s home.

Materials and Methods: Storage conditions of FM and FMF capsules have been tested for integrity and stability at ≤ -65 °C over a period of 12 months and at -15 °C ± 5 °C over a period of 3 months.

The composition of FM in the produced and stored bulk material should remain above 75 % similarity compared to the initial reference stool sample and should not drop below an acceptable 70 % similarity threshold predefined for the trial after 12 months of storage, as determined by 16S rRNA gene sequencing of the V1V2 variable regions of the 16S rRNA gene and Bray-Curtis similarity analysis.

The stability of FMF was proved by photometric analysis of bile acids, total protein, aspartate aminotransferase (ASAT) and glutamate dehydrogenase (GLDH) from 1 day up to 12 months after storage at ≤ -65 C°. The analytes must remain stable over this time as assessed using the Wilcoxon rank signed test.

The shipping process was validated by measuring the temperature in the transport boxes used (va-Q-med 7 premium, Va-Q-tec^®) at various time points up to 30 hours.

For the checking of IMP temperature during the transport, a temperature measurement strip was enclosed to the box, controlled and documented on an inspection sheet at the clinical trial centres to ensure temperature monitoring and storage conditions.

To assure the storage temperature at the patient's deep freezer, a thermometer is included with the delivery, allowing the patient to monitor the daily temperature of the IMP.

Results: When testing the microbiota stability of FM capsules the similarity of the bacterial community between the reference stool sample and the capsules stored at different temperatures (≤ -65 °C, -15 °C ± 5 °C) for different storage periods up to 12 month was > 78 % for all batches tested and thus above the accepted threshold.

Studies of FMF showed no negative impact of storage at ≤ -65 °C on the stability of the analytes (bile acids, total protein, ASAT, GLDH) in the FMF capsules over a period up to 12 months. Furthermore, an increased temperature of -15 °C ± 5 °C over 3 months also revealed no significant effect on stability of the analytes compared to storage at ≤ -65 °C.

The process validation for shipping the IMP was successfully completed. The shipping process with IMP was simulated and the temperature curve was monitored. After 24 hours, the temperature in the box was at least below -18.2 °C, and after 30 hours below -17 °C. The clinical trial centers confirmed the specified temperature of the IMP delivery and the storage conditions on the inspection sheet. All results met the specified acceptance criteria. The procedure and documentation of the shipping process were validated, justified, and approved for the FRESCO study.

The storage temperature of the IMP at the patient’s deep freezer is daily documented in the patient diary and proved by the study monitor.

Conclusion: Validated processes for storage and shipping of frozen IMPs within the FRESCO trial were established maintaining the integrity and stability of the IMPs throughout the entire supply chain. Based on the stability data collected, the Federal Institute for Drugs and Medical Devices (BfArM) approved an extension of the shelf life of the frozen capsules from 6 to 12 months. The processes were successfully carried out over a period of 180 IMP shipments, without any deviation in temperature.

Literatur

[1] Stallmach A, Grunert P , Stallhofer J, Löffler B, Baier M, Rödel J, Kiehntopf M, Neugebauer S, Pieper DH, Junca H, Tannapfel A, Merkel U, Schumacher U, Breternitz-Gruhne M, Heller T, Schauer A, Hartmann M, Steube A. Transfer of FRozen Encapsulated multi-donor Stool filtrate for active ulcerative Colitis (FRESCO): study protocol for a prospective, multicenter, double-blind, randomized, controlled trial. Trials. 2022 Feb 22;23(1):173. DOI: 10.1186/s13063-022-06095-1