Development of antiproliferative long-circulating liposomes co-encapsulating doxorubicin and curcumin, through the use of a quality-by-design approach

Abstract

The aim of this work was to use the quality-by-design (QbD) approach in the development of long-circulating liposomes co-loaded with curcumin (CUR) and doxorubicin (DOX) and to evaluate the cytotoxic potential of these liposomes in vitro using C26 murine colon carcinoma cell line. Based on a risk assessment, six parameters, namely the phospholipid, CUR and DOX concentrations, the phospholipid:cholesterol molar ratio, the temperature during the evaporation and hydration steps and the pH of the phosphate buffer, were identified as potential risk factors for the quality of the final product. The influence of these variables on the critical quality attributes of the co-loaded liposomal CUR and DOX was investigated: particle size, zeta potential, drug loading and entrapment efficiency. For this, a 26−2 factorial design was employed to establish a proper regression model and to generate the contour plots for the responses. The obtained data served to establish the design space for which different combinations of variables yielded liposomes with characteristics within predefined specifications. The validation of the model was carried out by preparing two liposomal formulations corresponding to the robust set point from within the design space and one outside the design space and calculating the percentage bias between the predicted and actual experimental results. The in vitro antiproliferative test showed that at higher CUR concentrations, the liposomes co-encapsulating CUR and DOX had a greater cytotoxic effect than DOX-loaded liposomes. Overall, this study showed that QbD is a useful instrument for controlling and optimizing the manufacturing process of liposomes co-loaded with CUR and DOX and that this nanoparticulate system possesses a great potential for use in colon cancer therapy.

Pharmaceutical Development of Liposomes Using the QbD Approach

“…Depending on its solubility, it can be entrapped in the aqueous core or in the lipid bilayer [40]. In order to achieve a greater therapeutic effect, more than one active substance may be encapsulated in the same product, so different physiochemical properties of one may influence the other/others drug entrapment process [40,41]. Besides this, their octanol/buffer partition must also be taken into consideration because a low partition will lead to a prolonged release of the drug and vice versa [42].…”

“…Regarding the preparation process, many techniques and methods like film hydration, emulsification, and reverse phase evaporation were developed but the most used one remains the film hydration method [37]. For this method to be efficient from the viewpoint of EE, parameters like temperature or rotation speed in the evaporation and hydration steps are critical and must be optimized [41,49]. Film hydration method's great disadvantage is that the obtained liposomes are in the vast majority of micron size with a multilamellar structure, being characterized by a high percentage of lipids and a reduced internal volume.…”

“…Regarding the drug concentration, several studies concluded that the total amount of drug used for encapsulation has a breaking point from which the EE cannot be further increased [41,46]. EE may also be influenced by PPs.…”

The effect of intravenous administration of liposomal curcumin in addition to sumatriptan treatment in an experimental migraine model in rats

“…Liposomal curcumin was characterized for mean particle size, polydispersity value, zeta potential, curcumin content, and encapsulation efficiency. 22,24 Thus, liposomal curcumin had a size around 140 nm; a low polydispersity value, close to 0.1; and a zeta potential of about −50 mV. The size, size distribution, and surface charge of the liposomes were determined immediately after preparation, upon dilution in a ratio of 1:100.…”

Effect of Liposomal Curcumin on Acetaminophen Hepatotoxicity by Down-regulation of Oxidative Stress and Matrix Metalloproteinases

“…Substances. Liposomal curcumin was encapsulated in longcirculating liposomes at a concentration of 4.7 mg/ml, using the film hydration method with a lipid molar ratio of 9.5:0.5:1 (1,2dipalmitoyl-sn-glycero-3-phosphocholine:1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000]-DPPC:PEG-2000-DSPE:CHO) as previously described (35,36). The proposed formulation had appropriate quality attributes for i.p.…”