Stasiłowicz-Krzemień Anna, Rosiak Natalia, Racaniello Giuseppe Francesco, Denora Nunzio, Cielecka-Piontek Judyta
Department of Pharmacognosy and Biomaterials, Poznan University of Medical Sciences, Rokietnicka 3 Str., 60-806 Poznan, Poland.
Department of Pharmacy-Pharmaceutical Sciences, University of Bari Aldo Moro, Orabona St. 4, 70125 Bari, Italy.
Int J Mol Sci. 2025 Aug 22;26(17):8126. doi: 10.3390/ijms26178126.
Apigenin (AP) is a natural flavonoid with senomorphic potential and neuroprotective action; however, poor aqueous solubility (<1 μg/mL) limits its bioavailability and therapeutic use. Therefore, the aim of this study was to obtain an amorphous dispersion of AP and evaluate its biological properties. Screening of AP solubilization capabilities under supercritical carbon dioxide processing conditions showed that the system with Soluplus (SOL) achieved the greatest improvement in AP dissolution (6455.4 ± 27.2 μg/mL). Using optimized process parameters (50 °C, 6500 PSI), the AP solubility increased to 8050.2 ± 35.1 μg/mL. X-ray powder diffraction (XRPD) confirmed amorphization, aligning with improved dissolution of AP in both acidic and neutral pH media. As a result, using the PAMPA model, an improvement in AP penetration through membranes simulating gastrointestinal and blood-brain barriers was demonstrated. The significant stability of the obtained amorphous AP dispersion (12 months at room conditions) was associated with stabilizing AP-solubilizer intermolecular interactions, mainly expressed as the shifts in the bands of AP in the range of 1018-1269 cm observed in ATR-FT-IR spectra. Chromatographic analysis confirmed the lack of AP decomposition immediately after the preparation of the amorphous dispersion, as well as after 12 months. As expected, the improvement of AP solubility is correlated with better biological activity assessed in selected in vitro tests such as antioxidant properties (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and cupric ion reducing antioxidant capacity (CUPRAC) assays) and anticholinesterase inhibition capabilities (AChE and BChE assays). The effect of the studies on improving AP solubility under supercritical carbon dioxide processing conditions is obtaining a stable amorphous AP dispersion (up to 12 months). Regardless of the pH of the media, an improvement in AP dissolution and penetration, conditioned by the passive diffusion process, through biological membranes was noted. Moreover, a more efficient antioxidant and neuroprotective effect of AP in the developed amorphous dispersion can also be suggested.
芹菜素（AP）是一种具有衰老形态潜力和神经保护作用的天然黄酮类化合物；然而，其较差的水溶性（<1μg/mL）限制了其生物利用度和治疗用途。因此，本研究的目的是获得AP的无定形分散体并评估其生物学特性。在超临界二氧化碳处理条件下对AP增溶能力的筛选表明，含有固体分散体辅料（SOL）的体系使AP的溶出度有最大程度的提高（6455.4±27.2μg/mL）。采用优化的工艺参数（50℃，6500磅力/平方英寸），AP的溶解度提高到8050.2±35.1μg/mL。X射线粉末衍射（XRPD）证实了非晶化，这与AP在酸性和中性pH介质中溶出度的提高一致。结果，使用平行人工膜渗透模型（PAMPA），证明了AP透过模拟胃肠道和血脑屏障的膜的渗透有所改善。所获得的无定形AP分散体在室温条件下具有显著的稳定性（12个月），这与稳定AP-增溶剂分子间相互作用有关，主要表现为在衰减全反射傅里叶变换红外光谱（ATR-FT-IR）中观察到的AP在1018-1269cm范围内的谱带位移。色谱分析证实，在制备无定形分散体后以及12个月后均未观察到AP分解。正如预期的那样，AP溶解度的提高与在所选体外试验（如抗氧化性能（2,2-二苯基-1-苦基肼自由基（DPPH）、2,2'-联氮-双-（3-乙基苯并噻唑啉-6-磺酸）（ABTS）和铜离子还原抗氧化能力（CUPRAC）测定））和抗胆碱酯酶抑制能力（乙酰胆碱酯酶（AChE）和丁酰胆碱酯酶（BChE）测定）中评估的更好的生物学活性相关。研究在超临界二氧化碳处理条件下提高AP溶解度的作用是获得了一种稳定的无定形AP分散体（长达12个月）。无论介质的pH值如何，均观察到AP的溶出度和透过生物膜的渗透有所改善，这是由被动扩散过程决定的。此外，还可以推测在开发的无定形分散体中AP具有更有效的抗氧化和神经保护作用。
