Department of Surgery, Duke University Medical Center, Durham, NC, USA.
Surgery. 2011 Aug;150(2):224-30. doi: 10.1016/j.surg.2011.05.015.
We report pharmacokinetic (PK) data, evaluation of modifications for increased stability, evaluation for cellular uptake, and mediation of regression of breast cancer for the aptamer OPN-R3.
The OPN-R3 aptamer was assessed for PK data in vivo with additional comparison of IV and subcutaneous dosing. Five aptamer variants were generated by differential 2'-O-methylation for comparison with parent. OPN-R3-Cy3 was incubated with MDA-MB231 cells and cellular uptake evaluated under confocal microscopy. Mice were treated with OPN-R3, mutant, or saline 3 weeks after inoculation with MDA-MB231 cells and tumor size was evaluated.
OPN-R3 PK data were: t(1/2) 7.76 hours, T(max) 3 hours, C(max) 13.2 mmol/L, mean residence time 9 hours, AUC (0-t) 161.9 mmol/hr/L, and K(d) 57.2 nmol/L. The half-life was higher when given intravenously versus subcutaneously (E(1/2) 7.93 vs 0.74 hours). The 2' methylation of all available bases increased unmodified aptamer stability and affinity (t(1/2) 6.2 hours; K(d) 520 nmol/L), but this did not improve on parent aptamer (t(1/2) 7.78 hours, K(d) 18 nmol/L). The aptamer remained extracellular. OPN-R3 caused regression of tumor to levels seen at 1 week after tumor inoculation.
We show the efficacy of OPN-R3 for reversing growth of breast cancer cells with adequate PK stability for clinical application.
我们报告了适体 OPN-R3 的药代动力学（PK）数据、增加稳定性的修饰评估、细胞摄取评估以及乳腺癌消退的介导作用。
评估了 OPN-R3 适体的体内 PK 数据，并与 IV 和皮下给药进行了额外比较。通过差异 2'-O-甲基化生成了 5 种适体变体，与亲本进行比较。将 OPN-R3-Cy3 与 MDA-MB231 细胞孵育，并在共聚焦显微镜下评估细胞摄取。在接种 MDA-MB231 细胞 3 周后，用 OPN-R3、突变体或生理盐水处理小鼠，并评估肿瘤大小。
OPN-R3 PK 数据为：t(1/2)7.76 小时，T(max)3 小时，C(max)13.2mmol/L，平均驻留时间 9 小时，AUC(0-t)161.9mmol/hr/L，和 K(d)57.2nmol/L。静脉内给药时半衰期高于皮下给药时（E(1/2)7.93 比 0.74 小时）。所有可用碱基的 2'甲基化增加了未修饰适体的稳定性和亲和力（t(1/2)6.2 小时；K(d)520nmol/L），但这并没有改善亲本适体（t(1/2)7.78 小时，K(d)18nmol/L）。适体仍然在细胞外。OPN-R3 导致肿瘤消退至接种后 1 周时的水平。
我们展示了 OPN-R3 逆转乳腺癌细胞生长的功效，其 PK 稳定性足以用于临床应用。
