利用Caco-2细胞模型研究新型Pim-1激酶抑制剂KY-C002的表观渗透系数

目的 利用人结肠癌Caco-2细胞模型研究新型Pim-1激酶抑制剂KY-C002的表观渗透系数,为药物制剂设计提供生物药剂学依据.方法 建立Caco-2细胞单层模型,通过细胞形态观察,跨上皮细胞电阻测定和酚红通透性实验验证Caco-2细胞单层完整性.考察不同浓度Pim-1激酶抑制剂KY-C002从绒毛面(AP侧)到基底面(BL侧),BL侧到 AP侧两个方向的转运情况.应用高效液相色谱法对该化合物进行定量分析,计算其表观渗透系数(Papp).结果 建立的Caco-2细胞单层完整,适用于转运实验.Pim-1激酶抑制剂KY-C002浓度为5、10、20 μg/ml时,Pim-1激酶抑制剂 KY-C002 从 AP 侧到 BL 侧的 Papp分别为(4. 72 ± 0.05) × 10-5、(5.29 ± 0.20) × 10-5、(2. 10 ± 0.41) × 10-5 cm/s(P<0.05);从BL侧到AP侧的Papp分别为(7.59±0.78)×10-6、(1.08 ± 0.04) × 10_5、(6.23 ± 0.63) × 10-6 cm/(P<0.05).结论 成功构建Caco-2细胞单层模型并用于考察Pim-1激酶抑制剂KY-C002的透过细胞单层的性能.在考察的浓度范围内,无论从AP侧到BL侧,还是从BL侧到AP侧,Pim-1激酶抑制剂KY-C002的Papp均与其浓度有关,浓度为10 μg/ml时的Papp大于5 μg/ml和20 μg/ml时的 Papp .

Study on cell permeability coefficients of a new Pim-1 kinase inhibitor KY-C002 by using Caco-2 cell monolayer

Objective To study the cell permeability coefficients of a new Pim-1 kinase inhibitor KY-C002 through a human colorectal cancer Caco-2 cell monolayer, providing a biological basis for the pharmacy pharmaceutical formulation design. Methods Caco-2 cell monolayer was built, and its integrity was confirmed through cell morphology observation, transepithelial electrical resistance measurement and Phenol Red Permeability experiment. The apparent permeability coefficients of Pim-1 kinase inhibitor KY-C002 from apical side (AP side) to basolateral side(BL side) or from BL side to AP side were investigated. The concentrations of KY-C002 were measured by high performance liquid chromatography. The apparent permeability coefficients (Papp) were then calculated. Results The Caco-2 cell monolayer was integrated. The Papp values of Pim-1 kinase inhibitor KY-C002 were (4. 72 ±0. 05)×10-5 cm/s, (5. 29 ±0.20) × 10-5 cm/s, (2. 10 ±0.41) × 10-5 cm/s (P <0.05) from AP side to BL side when the concentrations of KY-C002 were 5, 10, 20 μg/ml respectively. The Papp values of Pim-1 kinase inhibitor KY-C002 were (7.59 ± 0.78) × 10-6 cm/s, (1.08 ±0.04) × 10-5 cm/s, (6.23 ±0.63) × 10-6 cm/s (P < 0. 05) from BL side to AP side when the concentrations of KY-C002 were 5, 10, 20 μg/ml respectively. Conclusion The Caco-2 cell monolayer model was successfully constructed and used to investigate the properties of the permeated monolayer of Pim-1 kinase inhibitor KY-C002. The Papp of the Pim-1 kinase inhibitor KY-C002 was related to its concentration, and the Papp at the concentration of 10 μg/ml was greater than 5 μg/ml and 20 μg/ml of Papp.