载PROTAC分子白蛋白纳米粒的构建及对胶质瘤细胞NAD+代谢的抑制

目的 制备载蛋白降解靶向嵌合体（PROTAC）分子的白蛋白纳米粒并优化处方,考察纳米粒对胶质瘤细胞增殖活性及烟酰胺腺嘌呤二核苷酸（NAD⁺）代谢的抑制。方法 以热驱动法制备载NPT-B2的白蛋白纳米粒并表征,建立NPT-B2的高效液相色谱（HPLC）检测方法,以CCK8法和蛋白免疫印迹法分别考察NPT-B2和白蛋白纳米粒（B2-BSA-NPs）对胶质瘤细胞的增值活性抑制作用,及肿瘤细胞限速酶-烟酰胺磷酸核糖转移酶（NAMPT）的降解。结果 HPLC方法线性良好,精密度、回收率符合测定要求。纳米粒粒径为55.48 nm、电位-12.9 mV,包封率94.74%,载药量8.61%。NPT-B2对胶质瘤细胞的IC₅₀为61.16 nmol/L,同时具有NAMPT降解作用。B2-BSA-NPs对胶质瘤细胞的IC₅₀为41.21 nmol/L,对NAMPT具有非常显著降解效果。结论 构建了载PROTAC分子的白蛋白纳米粒,优化处方提高药物包封率,改善PROTAC分子水溶性差的问题,纳米粒对胶质瘤细胞增殖活性及NAD⁺能量代谢有显著抑制作用。

Construction of albumin nanoparticles loading PROTAC and its inhibition effect on NAD+ in glioma cells

Objective To prepare and optimize the formulation of Albumin nanoparticles loading PROTAC molecule and observe the inhibition effect of nanoparticles on the proliferation and NAD⁺ metabolism of glioma cells. Methods Albumin nanoparticles loading NPT-B2 were prepared and characterized with a thermal driving method, and the prescription was optimized. An HPLC method was established to determine the content of NPT-B2. The proliferation inhibition of NPT-B2 and B2-BSA-NPs on U251 cells were investigated by the CCK8 method, and the degradation effects of NPT-B2 and B2-BSA-NPs on NAMPT in glioma cells were investigated by western blotting. Results The HPLC method was stable, with good linearity, precision, and recovery rate. The nanoparticles had a particle size of about 55.48 nm, a potential of about -12.9 mV, an encapsulation rate of about 94.74%, and a drug loading amount of about 8.61%. The IC₅₀ of NPT-B2 on glioma cells was 61.16 nmol/L, which had a degradation effect on NAMPT. The IC₅₀ of B2-BSA-NPs on glioma was 41.21 nmol/L, which had a very significant degradation effect on NAMPT. Conclusion Albumin nanoparticles loading PROTAC molecules were constructed. The prescription was optimized to improve the drug encapsulation rate, and the low water solubility of PROTAC molecule was improved, which had a significant inhibitory effect on the proliferation and NAD⁺ energy metabolism of glioma cells.