穿心莲内酯总磺化物在细胞模型上的转运研究

Transport Study of Andrographolide Sulfonate on Cell Models

目的建立穿心莲内酯总磺化物反相高效液相色谱含量测定方法,研究其注射液中2种主要成分在calu-3和Caco-2细胞模型上的转运情况,评价将其应用于肺部给药和口服给药的安全性和有效性。方法细胞毒性实验筛选安全给药浓度。安全浓度以内的穿心莲内酯总磺化物注射液作用于Calu-3和Caco-2细胞模型,一定时间内取样,高效液相色谱法测定,考察浓度及时间对转运的影响,测定其主成分的表观渗透系数。结果药物浓度小于500μg·mL^(-1)时,Calu-3和Caco-2细胞存活率接近100%。建立的细胞模型显示出优良的紧密性和完整性。在Calu-3细胞模型上,17-氢-9-去氢穿心莲内酯-19-硫酸酯钠和17-氢-9-去氢穿心莲内酯双侧转运表观渗透系数均为1×10^(-5)数量级,在Caco-2细胞模型上,均为1×10^(-6)数量级。在2种细胞模型上,其外排比均小于2。结论穿心莲内酯总磺化物注射液主成分不是P糖蛋白的底物,在胃肠道中具有中等吸收,在呼吸道吸收良好,即肺部给药安全性和有效性更佳。

OBJECTIVE To establish a method for determination of the total content of andrographolide sulfonate, transport of two kinds of main components in the injection in Calu-3 and Caco-2 cell model, can be applied to the assessment of lung and oral administration to the safety and effectiveness of drugs. METHODS Screening of safe concentration of drugs by cytotoxicity test. Injection of andrographolide total sulfonate in safe concentrationwere applied to Calu-3 and Caco-2 cell models. Samples were taken for a certain period of time to determine the influence of concentration and time on the transport, and the apparent osmotic coefficients of the principal components were calculated. RESULTS When the concentration of the drug was less than 500 μg·mL-1, the survival rate of Calu-3 and Caco-2 cells was close to 100%.The established cell model showed excellent compactness and integrity.In the Calu-3 cell model, the apparent permeability coefficients of andrographolide sulfate C and 9-dehydro-17-hydro-andrographolide were 1×10-5 orders of magnitude.In the Caco-2 cell model, both were 1×10-6 orders of magnitude.On two cell models, the efflux ratios were less than 2. CONCLUSION The principal component of andrographolide total sulfonate injection is not the substrate for P glycoproteins, it has moderate absorption in the intestine, and has good absorption in the respiratory tract. Pulmonary administration is safer and more effective.