【摘要】：7-乙基-10-羥基喜樹堿(SN-38)是臨床上抗腫瘤藥伊立替康(CPT-11)的活性代謝產(chǎn)物,SN-38具有活性高、活性強(qiáng)的特點(diǎn),但一些因素限制了其臨床應(yīng)用,如SN-38水溶性差,口服生物利用度低,體內(nèi)消除較快。本文采用薄膜-探超法制備普通脂質(zhì)體,再以葉酸、殼聚糖為原料合成葉酸-殼聚糖偶聯(lián)物(FA-CS),然后通過吸附作用加載于脂質(zhì)體表面,制備成FA-CS包裹的脂質(zhì)體(FA-CS-SN-38-LP)。一方面,該制劑解決了藥物難溶于水的難題;另一方面該制劑具有緩釋和靶向的效果,可為SN-38的進(jìn)一步開發(fā)和應(yīng)用提供依據(jù)。通過單因素考察和正交試驗(yàn),確定SN-38脂質(zhì)體(SN-38-LP)、FA-CS包裹的脂質(zhì)體(FA-CS-SN-38-LP)的處方和工藝,并以制劑的粒徑、zeta電位、載藥量、包封率、體外釋放及穩(wěn)定性等為評(píng)價(jià)指標(biāo),對(duì)制劑的性質(zhì)進(jìn)行表征。優(yōu)化后,SN-38-LP和FA-CS-SN-38-LP的平均粒徑分別為158nm和239nm,Zeta電位分別為-7.15mV和㧏18.63m V。采用超濾-離心法測(cè)定脂質(zhì)體的包封率為79.23%,載藥量為3.85%。采用透析法研究體外釋放,比較SN-38-LP和FA-CS-SN-38-LP的釋放特性。結(jié)果表明,與SN-38溶液劑(SN-38-Sol)和SN-38--LP相比,FA-CS-SN-38-LP的釋放較慢,此外,SN-38-LP、FA-CS-SN-38-LP均屬于脂質(zhì)體載藥體系,體外釋放曲線相似。在體外細(xì)胞毒性研究中,以原料藥為對(duì)照,MTT法考察不同濃度制劑(SN-38-LP、FA-CS-SN-38-LP)對(duì)腫瘤細(xì)胞(MCF-7、HepG-2)的抑制作用。在給藥24h、48h、72h的情況下,FA-CS-SN-38-LP組對(duì)細(xì)胞的抑制作用均優(yōu)于SN-38-Sol、SN-38-LP組。細(xì)胞抑制程度與制劑中藥物的濃度成正相關(guān)。FA-CS-SN-38-LP對(duì)MCF-7細(xì)胞的抑制作用強(qiáng)于HepG-2細(xì)胞,該制劑對(duì)MCF-7細(xì)胞有較好抑制效果。建立S180荷瘤小鼠模型,考察制劑FA-CS-SN-38-LP在小鼠體內(nèi)的組織分布情況。在體內(nèi)抗腫瘤活性研究中,主要以重量抑瘤率和體積抑瘤率為評(píng)價(jià)指標(biāo)。實(shí)驗(yàn)結(jié)果顯示,FA-CS-SN-38-LP組重量抑瘤率和體積抑瘤率分別為67.04%和65.73%;與生理鹽水組相比,SN-38-LP組和FA-CS-SN-38-LP組腫瘤生長較慢,且腫瘤表面無潰爛、出血現(xiàn)象。病理學(xué)檢查結(jié)果顯示,相比生理鹽水組,FA-CS-SN-38-LP組腫瘤細(xì)胞排列疏松,呈不同程度的退變,腫瘤細(xì)胞數(shù)量減少,說明該制劑可較大程度地抑制腫瘤細(xì)胞的生長,具有較好的抗腫瘤效果。
[Abstract]:7-ethyl-10-hydroxycamptothecin (SN-38) is an active metabolite of the antitumor drug CPT-11. SN-38 has the characteristics of high activity and strong activity, but some factors limit its clinical application. For example, SN-38 has poor water solubility, low oral bioavailability and rapid elimination in vivo. In this paper, the common liposomes were prepared by thin-film method, then folic acid and chitosan were used as raw materials to synthesize folic acid-chitosan coupling compound (FA-CS), and then loaded on the surface of liposomes by adsorption. FA-CS encapsulated liposomes (FA-CS-SN-38-LP) were prepared. On the one hand, the preparation solves the problem that the drug is insoluble in water, on the other hand, it has the effect of slow release and targeting, which can provide the basis for the further development and application of SN-38. The formulation and process of SN-38 liposome (SN-38-LP) and FA-CS encapsulated liposome (FA-CS-SN-38-LP) were determined by single factor investigation and orthogonal test. The preparation size, zeta potential, drug loading capacity and encapsulation efficiency were determined. The properties of the preparation were characterized by in vitro release and stability. The average particle sizes of SN-38-LP and FA-CS-SN-38-LP were-7.15mV and-18.63 MV, respectively, and the average particle size of 158nm and FA-CS-SN-38-LP were-7.15mV and -18.63 MV, respectively. The entrapment efficiency of liposome was 79.23 by ultrafiltration-centrifugation, and the drug loading was 3.85. The release characteristics of SN-38-LP and FA-CS-SN-38-LP in vitro were studied by dialysis. The results showed that the release of FA-CS-SN-38-LP was slower than that of SN-38 solution (SN-38-Sol) and SN-38--LP. In addition, SN-38-LP,FA-CS-SN-38-LP belonged to liposome drug delivery system. In vitro release curve is similar. In vitro cytotoxicity study, the inhibitory effects of different concentrations of SN-38-LP,FA-CS-SN-38-LP on tumor cells (MCF-7,HepG-2) were investigated by MTT. The inhibitory effect of FA-CS-SN-38-LP group on cells was better than that of SN-38-Sol,SN-38-LP group at 24 h and 48 h / 72 h. The inhibitory effect of FA-CS-SN-38-LP on MCF-7 cells was stronger than that on HepG-2 cells, and the inhibitory effect of FA-CS-SN-38-LP on MCF-7 cells was better than that on HepG-2 cells. The model of S 180 tumor bearing mice was established to investigate the tissue distribution of FA-CS-SN-38-LP in mice. In the study of antitumor activity in vivo, the weight inhibition rate and volume inhibition rate were used as the evaluation indexes. The results showed that the weight inhibition rate and volume inhibition rate of FA-CS-SN-38-LP group were 67.04% and 65.73%, respectively. Compared with normal saline group, SN-38-LP group and FA-CS-SN-38-LP group had slower tumor growth, and no ulceration and bleeding on tumor surface. Pathological examination showed that compared with normal saline group, the tumor cells in FA-CS-SN-38-LP group were loosely arranged, with varying degrees of degeneration, and the number of tumor cells decreased, indicating that the preparation could inhibit the growth of tumor cells to a large extent. It has good antitumor effect.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R943

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本文編號(hào)：2307584