(W1030-01-05) Sustainable Development of a Nanosystem for the Co-encapsulation of Apigenin and Curcumin for Pharmaceutical Applications

Purpose: Developing innovative pharmaceutical systems that enhance conventional treatments with more sustainable or biodegradable raw materials to mitigate environmental impact draws attention to compounds from natural sources with therapeutic properties. Such is the case with apigenin and curcumin, which have individually demonstrated anti-inflammatory, antibacterial, and anticancer properties. This study aimed to design and optimize a sustainable nanostructured system for co-encapsulation of apigenin and curcumin, utilizing green solvents such as ethanol and renewable feedstock. This approach employs nanotechnology to craft sustainable drug delivery systems aimed at enhancing treatment efficacy and reducing toxic effects associated with conditions like breast cancer.

Methods: The nanostructured system of apigenin (API) and curcumin (CUR) was fabricated using the nanoprecipitation method with some modifications, employing ethanol:water as the solvent phase and pH 4 water as the antisolvent. Additionally, biocompatible, biodegradable, and renewable feedstock such as zein (encapsulating agent) and chitosan (coating agent) were utilized to achieve a fully biodegradable product. A Box-Behnken design was employed to optimize the independent variables: zein concentration, chitosan concentration, and API:CUR concentration. Response variables measured included particle size (PS), polydispersity index (PDI), zeta potential (ζ), and encapsulation efficiency (EE%) for each experimental batch. Once the nanosystem (NC API-CUR) was optimized, it was characterized through IR spectroscopy, DSC, and SEM morphology analysis. DDPH assay was conducted to explore potential NC API-CUR pharmaceutical applications and determine antioxidant capacity. A qualitative assessment of its antimicrobial activity against Staphylococcus aureus and Escherichia coli was compared with free API and CUR extracts.

Results: The modified nanoprecipitation method enabled co-encapsulation of API and CUR with nanometric sizes. According to desirability analysis (D=0.7178) derived from the Box-Behnken design, optimal conditions for manufacturing NC API-CUR were determined to be 1.2% w/v zein, 0.18% w/v chitosan, and 0.55 mg/mL API and 0.45 mg/mL CUR (Figure 1). Under these conditions, nanoparticles were obtained with a PS of 137.4 ± 6.1 nm, PDI of 0.300 ± 0.06, ζ of 50.1 ± 3.7 mV, and EE of 79.7% for API and 80.3% for CUR. A monodisperse, stable nanosystem with high encapsulation efficiency for simultaneous API and CUR administration was obtained. The suspension of nanoparticles exhibited a milky yellow coloration due to zein. IR and DSC results indicated the coupling of active principles, as evidenced by the decrease or disappearance of signals corresponding to individual extracts. The antioxidant activity assay revealed more significant DDPH radical inhibition for NC API-CUR, approximately 45%, compared to unencapsulated API-CUR at around 30% (Figure 2). Regarding antimicrobial activity, a more significant inhibition zone was observed for the API-CUR solution in the Staphylococcus aureus test compared to NC API-CUR, attributed to the time required for drugs to deliver the nanostructure. No inhibition zone was observed for both NC API-CUR and API-CUR solution in the Escherichia coli test (Figure 3).

Conclusion: This study successfully produced a fully biodegradable, monodisperse nanosystem with high encapsulation efficiency for the simultaneous administration of apigenin and curcumin. A more sustainable product was also obtained, utilizing only biodegradable, biocompatible materials derived from renewable sources with potential antioxidant and antimicrobial properties. These findings underscore the importance of developing products with natural drugs to prevent, control, and treat certain conditions.

References: Adel, M., Zahmatkeshan, M., Akbarzadeh, A., Rabiee, N., Ahmadi, S., Keyhanvar, P., & Seifalian, A. M. (2022). Chemotherapeutic effects of Apigenin in breast cancer: Preclinical evidence and molecular mechanisms; enhanced bioavailability by nanoparticles. Biotechnology Reports, 34, e00730.
Brotons-Canto, A., González-Navarro, C. J., Gil, A. G., Asin-Prieto, E., Saiz, M. J., & Llabrés, J. M. (2021). Pharmaceutics zein nanoparticles improve the oral bioavailability of curcumin in Wistar rats.
Cunha, I. V. N., Campos, A. M., & Caon, T. (2024). Nano-based apigenin delivery systems for cancer applications. Journal of Drug Delivery Science and Technology, 105334.
Hosseini-Zare, M. S., Sarhadi, M., Zarei, M., Thilagavathi, R., & Selvam, C. (2021). Synergistic effects of curcumin and its analogs with other bioactive compounds: A comprehensive review. European Journal of Medicinal Chemistry, 210.

Acknowledgements: Lizbeth Martínez-Acevedo and Moises Galindo-Pérez acknowledge the Consejo Nacional de Humanidades Ciencias y Tecnologías (CONAHCyT) for the postdoctoral scholarship awarded.


Figure 1. The response surface as a function of a) PS; b) PDI; c) ζ; d) EE% CUR; e) EE% API.



Figure 2. Antioxidant activity of NC API-CUR compared with unencapsulated API and CUR solutions.



Figure 3. Antimicrobial activity of NC API-CUR compared with unencapsulated API and CUR solutions.