载紫杉醇的聚乙二醇修饰的大黄酸偶联物胶束的体内安全性及抗肿瘤药效学研究

Study on in Vivo Safety and Anti-tumor Pharmacodynamics of Paclitaxel-loaded PEGylated Rhein Conjugate Micelles

关键词:  紫杉醇  聚合物胶束  大黄酸偶联物  抗肿瘤

Key words:  paclitaxel  polymer micelles  rhein conjugate  anti-tumor

摘要:

目的 探讨载紫杉醇(paclitaxel，PTX)的聚乙二醇修饰的羧甲基壳聚糖-大黄酸(CRmP)偶联物胶束(PTX/CRmP胶束)作为静脉注射给药制剂的安全性，并对PTX/CRmP胶束在小鼠体内的抗肿瘤效果进行研究。方法 测定PTX/CRmP胶束的pH值及渗透压。以新鲜兔血红细胞检测溶血性。新西兰白兔耳缘静脉注射PTX/CRmP胶束溶液、CRmP偶联物溶液，光镜下对血管连同周围组织切片进行组织学检查。建立H-22荷瘤小鼠模型，随机分组，分别尾静脉注射给予0.9%氯化钠注射液、注射用PTX脂质体、PTX注射液和PTX/CRmP胶束低、中、高剂量(5，10，15 mg·kg^-1)。给药过程中每天观察小鼠活动情况，记录小鼠体质量变化、肿瘤体积，绘制体质量变化曲线、肿瘤体积变化曲线，计算抑瘤率，HE染色观察肿瘤组织切片。结果 不同浓度PTX/CRmP胶束溶液的pH值与血液pH值相近，渗透压为286~292 mOsm·kg^-1。浓度在0.01~0.05mg·mL^-1内，CRmP偶联物及PTX/CRmP胶束的溶血率<5%；经耳缘静脉注射后，注射部位肉眼可见血管无红肿、充血等刺激性现象，血管均未见到明显的病理变化。PTX/CRmP胶束能抑制肿瘤生长和转移，抑制效果呈现剂量依赖。当给药剂量为10 mg·kg^-1时，PTX/CRmP胶束的抑瘤率达到了52.11%，抑瘤效果优于注射用PTX脂质体与PTX注射液。PTX/CRmP胶束组小鼠体质量增长，肿瘤无明显的全身扩散现象，精神状态良好。病理组织切片表明，PTX/CRmP胶束高剂量组肿瘤组织坏死程度最大。结论 PTX/CRmP胶束具有良好的生物安全性，用于静脉注射给药可有效抑制肿瘤生长，同时可降低PTX的不良反应，提高PTX抗肿瘤效果，为临床应用提供参考。

Abstract:

OBJECTIVE To investigate the safety of paclitaxel(PTX)-loaded PEGylated carboxymethyl chitosan-rhein (CRmP) polymeric micelles(PTX/CRmP PMs) as intravenous drug delivery preparations, and to investigate the anti-tumor effect of PTX/CRmP PMs in mice. METHODS The pH value and osmotic pressure of PTX/CRmP PMs were measured. The hemolysis was detected using the blood cells of New Zealand Rabbit. New Zealand rabbits were injected with PTX/CRmP PMs and CRmP conjugate solutions via the auricle vein. The vessels along with surrounding tissue sections were examined histologically under light microscopy. H-22 tumor-bearing mice models were established and divided into 0.9% sodium chloride injection group, paclitaxel liposome for injection group, paclitaxel injection group, PTX/CRmP PMs groups in low, medium and high dose(5, 10, 15 mg·kg^-1). The mice were administered via tail vein. During administration, the activity was observed every day. Changes of body weight and tumor volume were measured. The body weight change curve and tumor inhibition rate were plotted. Morphological changes in tumor tissues were observed by HE staining. RESULTS The pH values of PTX/CRmP PMs at different concentrations were similar to blood, with an osmotic pressure of 286-292 mOsm·kg^-1. In the concentration range of 0.01~0.05 mg·mL^-1 the hemolysis of CRmP conjugate and PTX/CRmP PMs were <5%. After intravenous injection via the auricle vein, no irritation phenomena were observed in the blood vessels at the injection site, and no obvious pathological changes were observed in the blood vessels. Tumor growth and metastasis were inhibited in PTX/CRmP PMs group, and the tumor-inhibiting effect showed dose-dependent. At the administered dose was 10 mg·kg^-1, the tumor inhibition rate of PTX/CRmP PMs reached 52.11%, better than that of paclitaxel liposome for injection group and paclitaxel injection group. Moreover, the mice in the PTX/CRmP PMs group gained weight, and no apparent whole-body diffusion was observed at the tumor site. During the treatment, the mice were in a good mental state. Pathological tissue sections indicated that the degree of tumor tissue necrosis was the greatest in the PTX/CRmP PMs high-dose group. CONCLUSION PTX/CRmP PMs have good biosafety and can be used for intravenous administration to effectively inhibit tumor growth, while reducing the toxic side effects of PTX and improving the anti-tumor effect of PTX, providing a reference for clinical application.