Quality by Design Approach in Liposomal Formulations: Robust Product Development

Nanomedicine is an emerging field with continuous growth and differentiation. Liposomal formulations are a major platform in nanomedicine, with more than fifteen FDA-approved liposomal products in the market. However, as is the case for other types of nanoparticle-based delivery systems, liposomal formulations and manufacturing is intrinsically complex and associated with a set of dependent and independent variables, rendering experiential optimization a tedious process in general. Quality by design (QbD) is a powerful approach that can be applied in such complex systems to facilitate product development and ensure reproducible manufacturing processes, which are an essential pre-requisite for efficient and safe therapeutics. Input variables (related to materials, processes and experiment design) and the quality attributes for the final liposomal product should follow a systematic and planned experimental design to identify critical variables and optimal formulations/processes, where these elements are subjected to risk assessment. This review discusses the current practices that employ QbD in developing liposomal-based nano-pharmaceuticals

Fig 4 -

A) Induced apoptosis of MCF-7_WT cells treated with Dis and Hyd alone and combined B) Induced apoptosis of MCF-7_DoxR cells treated with Dis and Hyd alone and combined.</p

STRING analysis of protein–protein interactions.

(A) Interaction between proteins involved in drug resistance STAT3, NOTCH, DNMT, (B) Illustration of combination of HDACi and DNMTi pathway, (C) combination of HDACi and DNMTi and the effect of cell apoptosis proteins, (D) interactions of DNMTi and proteins ivoved in cell cycle, (E) Effect of ALDH enzymes on P53 proteins, (F) Interactions between ALDH and glutathione (GSH), (G) Interactions between ALDH, ABCC1 and p53 proteins, (H) Signalling pathway showed interaction of DNMT, ALDH, P53, TOP2A, ABCB1. STAT3, NOCH1. (thickness of edges indicate confidence).</p

Diagnostic graphics produced for synergistic affect quantification.

(A) Isobologram for Dis/Hyd combination at different concentration points, (B) The Fa-DRI plot (Chou-Martin plot) for the non-constant ratios of Dis/Hyd combination, (C) The fraction affected (Fa) versus combination index (CI) plot after treatment with Dis/Hyd combination, that most of CI values are 50 of Dis, Hyd and Dis/Hyd combination.</p

Diagnostic graphics produced for synergistic effect quantification of DOX/Dis/Hyd combination against MCF-7_DoxR cells.

(A) Dose -Fa curve for DOX alone and for DOX with (DOX/Dis/Hyd combination) at different concentration points (B) Table of IC50 of DOX, Dis, Hyd alone and combination (C) The fraction affected (Fa) versus combination index (CI) plot after treatment with DOX/Dis/Hyd combination, that most of CI values are < 1 for the range of 0.25–0.8 (D) The Fa-DRI plot for the non-constant ratios of DOX/Dis/Hyd combination.</p

Apoptosis Induced by DOX/Dis/Hyd single treatment and combinations.

A) and B) MCF-7_WT cells, C) and D) MCF-7_DoxR cells.</p

Fig 7 -

IC50 values after treatment with DOX/Dis/Hid. The MCF-7_WT and MCF-7_DoxR cells were treated with DOX, Dis, Hyd and DOX/Dis/Hyd to assess the cytotoxicity levels (A) The dose-response curve for cells treated with Hyd and; (B) The dose-response curve for cells treated with Dis; (C)The dose-response curve for cells treated with DOX; (D) The dose-response curve for cells treated with DOX/Dis/Hyd in combination, the curve showing DOX IC50 with and without combination with Dis/Hyd (0.03/20 μM). All cytotoxicity values represent the average ± SD of three independent experiments.</p

Fig 1 -

The chemical structures of disulfiram (A), hydralazine (B), and doxorubicin (C).</p

Diagnostic graphics produced for synergistic effect quantification of DOX/Dis/Hyd combination against MCF-7_WT cells.

(A) Dose -Fa curve for DOX alone and for DOX with (DOX/Dis/Hyd combination) at different concentration points (B) Table of IC50 of DOX, Dis, Hyd alone and combination (C) The fraction affected (Fa) versus combination index (CI) plot after treatment with DOX/Dis/Hyd combination, that most of CI values are <1 for the range of 0.5–0.95(D) The Fa-DRI plot for the non-constant ratios of DOX/Dis/Hyd combination.</p