【摘要】：目的：多西他賽(docetaxel, DTX)是一種新型植物堿類抗腫瘤藥,為了克服其在水中溶解度低的缺點,制備了一種能夠包載多西他賽的聚乙二醇-聚乳酸mPEG-b-PDLLA共聚物；并且對多西他賽載藥納米膠束的進行了體內(nèi)外評價。方法：以聚乙二醇單甲醚mPEG和D,L-丙交酯為原料,在辛酸亞錫的催化下,開環(huán)聚合,形成嵌段聚合物。通過傅立葉變換紅外光譜(FT-IR)、核磁共振(1H-NMR)、差示掃描量熱法(DSC)、 MALDI-TOF-TOF-MS、氣相色譜法、電感耦合法、pH計等對聚合物mPEG-b-PDLLA進行了表征。采用溶劑揮發(fā)薄膜水化法制備了包載抗癌藥物多西他賽的納米膠束(DTX-PM)。利用Malvern Nano ZS激光粒度分析儀測定其粒徑和分布；透射電鏡觀測納米膠束的形態(tài)；高效液相色譜法(HPLC)測定包封率及載藥量,并進行了質(zhì)量標準研究；通過粒徑和載藥量的測定觀察載多西他賽納米膠束的穩(wěn)定性。通過細胞遷移試驗、MTT法和激光共聚焦實驗考察載多西他賽納米膠束對A549細胞的增殖抑制作用。裸鼠皮下接種瘤塊建立肺癌動物模型后,采用尾靜脈的方法分別給裸鼠注射葡萄糖水溶液、DTX-PM溶液及市售多帕菲(?)溶液,定期觀察裸鼠皮下瘤體體積大小和裸鼠體重的變化。結(jié)果：通過1H-NMR得出丙交酯的摩爾百分含量為17～19%, mPEG2000的摩爾百分含量為81%～83%,共聚物平均分子量在3300左右。通過紅外光譜譜圖的分析證實了丙交酯和mPEG聚合形成了嵌段共聚物。MALDI-TOF-TOF-MS實驗顯示檢測峰主要在3300 Da,純化后雜質(zhì)峰明顯減少。DSC測試得到mPEG-b-PDLLA聚合物的熔點約為41.8℃,熔融焓Hm=20.3 mJ/mg。殘留量檢查的結(jié)果顯示甲苯、二氯甲烷、乙醚等有機溶劑并無殘留,丙交酯的殘留含量為1.2%,錫的殘留含量為1.521E+05μg/kg。 mPEG-b-PDLLA溶于水后測得pH值為5.2。多西他賽納米膠束的粒徑約16 nm左右,PDI在0.1～0.2之間；透射電鏡下觀察到DTM-PM的形狀呈規(guī)則的球形。通過HPLC檢測,得出多西他賽納米膠束的包封率為(88.88±0.61)%,載藥量為(15.00±0.69)%。質(zhì)量標準研究中的含量測定方法簡便、準確度高、重現(xiàn)性好�？疾霥TX-PM的復溶穩(wěn)定性,結(jié)果顯示膠束穩(wěn)定性良好,可以滿足輸液要求。MTT法結(jié)果顯示,高濃度時市售多帕菲(?)比DTX-PM具有更強的細胞增殖抑制作用,而在低濃度時DTX-PM具有更強的細胞增殖抑制作用；細胞遷移試驗顯示,市售多帕菲(?)和DTX-PM均可抑制A549細胞的增殖；激光共聚膠試驗表明,膠束的入胞速度很快且能夠攝取進入細胞核。給裸鼠注射葡萄糖溶液、市售多帕菲(?)溶液和DTX-PM溶液后發(fā)現(xiàn)DTX-PM組的裸鼠腫瘤生長速度慢于市售多帕菲(?)組,且毒性小于市售多帕菲(?)組。結(jié)果表明,DTX-PM的抗肺癌作用強于現(xiàn)在臨床上使用的多帕菲(?)。結(jié)論：通過mPEG-b-PDLLA共聚物的結(jié)構(gòu)表征,可以證實丙交酯和mPEG成功聚合成了嵌段聚合物。聚合物中有機溶劑、丙交酯和金屬錫的殘留量均符合要求。載多西他賽納米膠束的形態(tài)呈球形,其粒徑分布均勻,性質(zhì)穩(wěn)定。質(zhì)量標準研究中的含量測定方法簡便、準確度高、重現(xiàn)性好,可以用于該制劑的質(zhì)量控制。多西他賽納米膠束能夠有效地抑制A549細胞的增殖,具有抗肺癌作用。
[Abstract]:Objective: To overcome the disadvantage of low solubility in water, a new type of anti-tumor medicine of plant base is prepared by using Dosizel (DTX), and a polyethylene glycol-polylactic acid (PEG-b-PDLLA) copolymer which can be used for carrying out the polysiracite is prepared. and in vivo out-of-vivo evaluation of the nano-micelle of the multi-sizinese medicament-carrying agent. Methods: The block polymer was formed by ring-opening polymerization under the catalysis of stannous octoate as the raw material with polyethylene glycol monomethyl ether (mPEG) and D (L-lactide). The polymer mPEG-b-PDLLA was characterized by Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (1H-NMR), differential scanning calorimetry (DSC), MALDI-TOF-TOF-MS, gas chromatography, inductive coupling method, pH meter and the like. The nano-micelle (DTX-PM) was prepared by the method of solvent volatilization film hydration. The particle size and distribution were determined by a Malvern Nano-ZS laser particle size analyzer, the morphology of the nano-micelle was observed by transmission electron microscope, the encapsulation rate and the drug-carrying amount were determined by high performance liquid chromatography (HPLC), and the quality standard was studied. The stability of the nano-micelle is observed by the determination of the particle size and the drug-loading amount. The proliferation of A549 cells was studied by cell migration test, MTT method and laser confocal experiment. After a lung cancer animal model was established by subcutaneous inoculation of nude mice, a method of tail vein was used to inject glucose solution, DTX-PM solution and multi-pafenone (?) in the nude mice, respectively.) The volume size of the subcutaneous tumor of the nude mice and the body weight of the nude mice were observed on a regular basis. Results: By 1H-NMR, the molar percentage content of the acrylate was 17-19%, the mole percentage of mCloth was 81-83%, and the average molecular weight of the copolymer was about 3300. Through the analysis of the infrared spectrum, the block copolymer was formed by the polymerization of the acrylate and the mPEG. MALDI-TOF-TOF-MS showed that the detection peak was mainly at 3300 Da, and the impurity peak after purification was significantly reduced. The melting point of the mPEG-b-PDLLA polymer was about 41. 8 鈩，

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