Robust freeze-drying process re-design of a legacy product based on risk analysis and design of experiments.

In this study, a QbD freeze-drying process re-design applied to a lyophilized injectable drug product is presented. The main objective was to assess the freeze-drying process robustness using risk analysis and a proper experimental design. First, the product's thermal fingerprint was characterized by thermal analysis and freeze-drying microscopy. Then, according to the output of the risk analysis, primary drying temperature and pressure were studied by a Doehlert DoE design with four responses; primary drying time, appearance, residual moisture content, and reconstitution time. Statistically significant MLR models were obtained for residual moisture content and primary drying time. In the latter, the temperature factor was the predominant factor to predict the duration of the primary drying stage. Two additional lab-scale batches were run to confirm the mathematical model predictions. Finally, optimal primary drying conditions (30 °C, 0.400 mbar) were selected to minimize the duration of the primary drying stage, while preserving the quality of the product. It was possible to set high temperature and pressure values because no collapse temperature was found during the thermal characterization of the product. Secondary drying temperature and time were defined based on the residual moisture content results. It was shown that secondary drying is robust between 30 °C and 50 °C and from 3 to 16 h. In conclusion, we were able to define a robust freeze-drying process which was further validated at an industrial scale with satisfactory results and approved by the health authorities in different countries around Europe.

Finding a reliable limit of detection in the NIR determination of residual moisture in a freeze-dried drug product.

The validation of an analytical method in the pharmaceutical industry follows strictly regulated guidelines. The introduction of multivariable calibration methods requires a revision of these recommendations, since some of them are contradictory regarding the limit of detection (LOD). This work compares the LOD values obtained using pseudounivariate and multivariate procedures in the PLS-NIR determination of residual moisture content (RMC) in a freeze-dried drug. As NIR has proved to be more precise than Karl-Fischer at low RMC values, LOD has been estimated by ordinary and by orthogonal least squares regression. The precision of the RMC determination in approx. 2000 industrial vials was used as an indirect evidence of the reliability of the LOD values obtained. The effect of reducing the number of calibration samples and increasing the RMC values have also been studied. No significant differences were observed using a number of calibration samples ≥ 20. Based on our findings, when the size of the calibration sample set is high and the range of RMC values is close to the limit, the LOD estimated with the ICH formula and using orthogonal regression should be recommended. If water content moves away, the ICH formula should be replaced by the LODOS equation as a practical, reliable and simple procedure.

Nanostructured lipid carriers for triamcinolone acetonide delivery to the posterior segment of the eye.

Triamcinolone acetonide (TA) is a corticosteroid drug currently administered by intravitreal injection for a broad spectrum of inflammatory, edematous and angiogenic ocular diseases. To increase the drug's bioavailability by ocular instillation, TA was encapsulated in nanostructured lipid carriers (NLC), previously optimized by our group using a factorial design approach. In the present paper, nanometric (∼200 nm), unimodal and negatively charged NLC loaded with the fluorescent lipid marker Nile red (NR-NLC) and drug (TA-NLC) were produced by high pressure homogenization. Based on the selected formulations, in vivo tests were carried out by eye-drop instillation of NR-NLC in mice, revealing the systems' ability of delivering lipophilic actives to the posterior segment of the eye via the corneal and non-corneal pathways. Short and long-term stability of TA-NLC was assessed by high performance stability analysis using the Turbiscan®. The results showed a backscattering of less than 1.5% and during a period of 6 months, anticipated the low tendency of these particles for aggregation during shelf life when stored at room temperature.