Key Laboratory of Drug Targeting and Drug Delivery Systems, West China School of Pharmacy, Sichuan University , Chengdu, 610041, China.
College of Polymer Science and Engineering, Sichuan University , Chengdu, 610065, China.
Biomacromolecules. 2017 Mar 13;18(3):865-876. doi: 10.1021/acs.biomac.6b01756. Epub 2017 Feb 27.
To alleviate the hemorrhagic side effect of thrombolysis therapy, a thrombus targeted drug delivery system based on the specific affinity of Annexin V to phosphatidylserine exposed on the membrane surface of activated platelet was developed. The amphiphilic and biodegradable biomaterial, polycaprolactone-block-poly(2-(dimethylamino)ethyl methacrylate)-block-poly(2-hydroxyethyl methacrylate) (PCL-b-PDMAEMA-b-PHEMA (PCDH)) triblock polymer, was synthesized via ring opening polymerization (ROP) and atom transfer radical polymerization (ATRP) to use as the nanocarriers of thrombolytic drug. In order to conjugate Annexin V to the polymer, PCDH was modified by succinic anhydride via ring-opening reaction to introduce the carboxyl group (PCDH-COOH). After preparation of PCDH/PCDH-COOH (9/1, m/m) mixed micelles, Annexin V was coupled with the micelles using carbodiimide chemistry. The blood clot lysis assay in vitro confirmed that lumbrokinase-loaded targeted micelles (LKTM) had stronger thrombolysis potency than free lumbrokinase (LK) and LK-loaded nontargeted micelles (LKM, P < 0.05). In vivo thrombolytic assay, multispectral, optoacoustic tomography (MSOT) was used to assess the target ability of LKTM. The results of MSOT images indicated the fluorescence intensity of the LKTM group located in the blood clot position were significantly stronger than the LKM group. A 5 mm of carotid artery containing blood clot was cut out 24 h later after administration to assess the degree of thrombolysis. The results of thrombolytic assay in vivo were consistent with the assay in vitro, which the differences between LK, LKM, and LKTM groups were both statistically significant. All the results of thrombolysis assays above proved that the capacity of thrombolysis in the LKTM group was optimal. It suggested that Annexin V-conjugated micelles will be a potential drug delivery system for targeted thrombolysis.
为了缓解溶栓治疗的出血副作用，开发了一种基于膜表面活化血小板暴露的磷脂酰丝氨酸与 Annexin V 特异性亲和力的血栓靶向药物输送系统。两亲性和可生物降解的生物材料聚己内酯嵌段聚（2-（二甲氨基）乙基甲基丙烯酸酯）嵌段聚（2-羟乙基甲基丙烯酸酯）（PCL-b-PDMAEMA-b-PHEMA（PCDH））三嵌段聚合物通过开环聚合（ROP）和原子转移自由基聚合（ATRP）合成，用作溶栓药物的纳米载体。为了将 Annexin V 连接到聚合物上，通过开环反应用琥珀酸酐修饰 PCDH 以引入羧基（PCDH-COOH）。制备 PCDH/PCDH-COOH（9/1，m/m）混合胶束后，用碳二亚胺化学法将 Annexin V 偶联到胶束上。体外血块溶解试验证实，载有蚓激酶的靶向胶束（LKTM）比游离蚓激酶（LK）和载有非靶向胶束（LKM，P <0.05）具有更强的溶栓活性。体内溶栓试验，多光谱，光声断层扫描（MSOT）用于评估 LKTM 的靶向能力。MSOT 图像的结果表明，位于血块位置的 LKTM 组的荧光强度明显强于 LKM 组。给药 24 小时后取出含有血块的 5 毫米颈动脉以评估溶栓程度。体内溶栓试验的结果与体外试验一致，LK、LKM 和 LKTM 组之间的差异均具有统计学意义。所有溶栓试验的结果均表明 LKTM 组的溶栓能力最佳。这表明 Annexin V 缀合的胶束将成为一种有前途的靶向溶栓药物输送系统。
