PP199. Investigation of antialzheimer and antiangiogenic activities of donepezil hydrochloride loaded PLGA nanoparticle (NP) systems by in vitro AchE and BuChE inhibitor activity analyzes and in vivo CAM method

Kömür M1, Kıyan H2, Öztürk A1
1Anadolu University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Eskişehir, Turkey, 2Anadolu University, Faculty of Pharmacy, Department of Pharmacognosy, Eskişehir, Turkey

Introduction: NPs with unique physicochemical and biological properties, are taken more easily by the cells than larger molecules; therefore, they can be used effectively as a drug delivery system (DDS). The most important reasons for the use of NPs as a DDS include prolonged release of encapsulated drug, increasing drug's therapeutic efficacy and reducing side effects [1]. For this purpose, PLGA-NPs were prepared in this study to increase the effectiveness of donepezil hydrochloride (DNP).

Methods: DNP loaded PLGA-based-NPs were prepared by the 'Double Emulsification Solvent Evaporation' method. The I-DNP coded formulation was selected as optimum and solid-state characterizations were performed with DSC, FT-IR and 1H-NMR [1]. After the characterization of I-DNP in terms of pharmaceutical technology (particle size, PDI, zeta potential, EE% and release) was completed, the Ellman Test was used to determine the acetylcholinesterase (AchE) and butyrylcholinesterase (BuChE) enzyme inhibition activity. In vivo CAM assay was used to study the effect of I-DNP on angiogenesis [1,2].

Results: Encapsulation was proven by DSC, FT-IR, and 1H-NMR. According to the AChE and BuChE inhibitor activity results, I-DNP showed strong anti-AChE activity with 96.88±0.16% inhibition at 400 μg/mL concentration and strong anti-BuChE activity with 82.96±0.57% inhibition at a concentration of 400 μg/mL. According to the in vivo CAM results, I-DNP showed anti-angiogenic effect by significantly inhibiting capillary vessel development at a concentration of 10 μg/pellet, compared to the blank (2.5% agarose) and I-Blank coded formulation. Depending on the decrease in capillary area, the activity order was determined as I-DNP˃DNP˃Blank formulation. Total capillary area was determined as 14.702%±3.395, 17.982%±2.595, and 20.362%±3.353 for I- DNP, DNP, and blank formulation, respectively.

Conclusion: According to the anti-alzheimer activity results, I-DNP showed strong anti-AChE and anti-BuChE activities compared to DNP, which is an acetylcholinesterase inhibitor. Within vivo CAM experiments, it was determined that the I-DNP coded formulation attenuated (reduced/inhibited) angiogenesis and it was concluded that the prepared NP system could be a new anti-angiogenic treatment by stimulating the cholinergic system. This study was financed by Anadolu University Scientific Research Project Foundation (No:2304S027).

[1] Öztürk AA, Kıyan HT. Treatment of oxidative stress-induced pain and inflammation with dexketoprofen trometamol loaded different molecular weight chitosan nanoparticles: Formulation, characterization and anti-inflammatory activity by using in vivo HET-CAM assay. Microvasc Res. 2020;128:103961.

[2] Kıyan HT, Demirci B, Başer KHC, Demirci F. The in vivo evaluation of anti-angiogenic effects of Hypericum essential oils using the chorioallantoic membrane assay. Pharm Biol. 2014;52(1):44-50.