本文選題：2-甲氧基雌二醇 + 牛血清白蛋白��； 參考：《鄭州大學》2014年碩士論文

【摘要】：2-甲氧基雌二醇(2-methoxyestradiol,2-ME)是一種甾體激素類化合物，屬新型激素類抗腫瘤藥物，對快速生長的腫瘤細胞具有相對較高的特異性，且不會對骨髓和造血器官產生毒性。由于2-ME是一種難溶性藥物，胃腸道吸收差，血漿消除率快，生物利用度低等特點，限制了其在臨床上的開發(fā)應用。因此本文以牛血清白蛋白(BSA)作為藥物載體，小分子葉酸(FA)作為靶向因子，抗腫瘤藥物2-甲氧基雌二醇(2-ME)作為模型藥物，構建成以葉酸受體介導的2-ME白蛋白納米粒腫瘤靶向給藥系統(tǒng)(2-ME-FA-BSANPs)，旨在提高2-ME的水溶性和生物利用度，減少給藥劑量，降低藥物毒副作用。 1.2-ME-FA-BSANPs的制備及相關性質的考察。 首先采用去溶劑化-化學交聯(lián)法制備2-ME白蛋白納米粒(2-ME-BSANPs)，通過單因素考察白蛋白濃度、2-ME濃度、交聯(lián)劑戊二醛用量、乙醇與水比例、溶液pH等條件，確定了2-ME-BSANPs制備工藝為：BSA濃度為40mg/ml；2-ME濃度為4mg/ml；溶液pH值為10.0；乙醇與水的比例為3:1。進一步利用葉酸活性酯與白蛋白納米粒表面活性氨基進行偶聯(lián)反應，制備葉酸偶聯(lián)的2-ME白蛋白納米粒靶向給藥系統(tǒng)(2-ME-FA-BSANPs)，使其具有主動靶向作用，并提高白蛋白納米粒的腫瘤細胞攝取率。 測定2-ME-FA-BSANPs平均粒徑和電位分別為(217.5±5.1) nm和(-37.59±1.39) mV；平均包封率與載藥量分別為(82.50±5.80)%和(7.50±0.50)%；掃描電鏡圖顯示:2-ME-FA-BSANPs粒徑分布均勻，呈球形且表面光滑；體外釋藥結果顯示：原料藥2-ME釋放較快，在12h內累積釋放量達85%左右；相對應的2-ME-FA-BSANPs釋放緩慢平穩(wěn)，在96h內累積釋放量達85%左右，表明靶向給藥系統(tǒng)具有一定的緩釋特性；將靶向制劑制備成凍干粉，經篩選選擇3%的蔗糖作為其凍干保護劑；考察穩(wěn)定性結果顯示：在4℃和室溫條件下凍干粉可穩(wěn)定保存6個月，表明該靶向制劑具有較好的穩(wěn)定性。 2.2-ME-FA-BSANPs體外抗腫瘤活性與靶向性的研究。 采用MTT法考察靶向給藥系統(tǒng)對SMMC-7721腫瘤細胞存活率的影響。在2-ME-FA-BSANPs作用時間24h、48h和72h，最大濃度為40μmol/l時，對應的腫瘤細胞抑制率分別為50.58%、68.53%和75.00%，相同條件下2-ME組的腫瘤細胞抑制率分別為25.28%、30.31%和35.37%，結果表明在不同時間、不同濃度作用下，2-ME-FA-BSANPs組對腫瘤細胞的抑制率均高于2-ME組(P0.05)。 采用熒光顯微鏡法與細胞流式分析法考察2-ME-BSANPs和2-ME-FA-BSANPs的靶向性。結果顯示：相比于2-ME-BSANPs組，，2-ME-FA-BSANPs組的熒光強度明顯增強，且在作用4h時2-ME-FA-BSANPs組進入細胞內的比例為99.8%，表明靶向給藥系統(tǒng)在葉酸介導作用下可以有效、快速的進入腫瘤細胞內部，對腫瘤細胞具有較高的靶向作用；細胞周期研究結果表明所制備靶向給藥系統(tǒng)主要將SMMC-7721腫瘤細胞阻滯在G2/M期，且不改變原料藥2-ME的作用機制；細胞凋亡研究結果表明，與2-ME組相比，靶向給藥系統(tǒng)2-ME-FA-BASNPs能誘導更多腫瘤細胞發(fā)生凋亡。 3.2-ME-FA-BSANPs體內藥代動力學的研究。 本研究以Sprague-Dawle(SD)大鼠為動物模型，采用HPLC法測定2-ME在大鼠血漿中的濃度，研究靶向給藥系統(tǒng)的體內藥代動力學特征。結果顯示，2-ME-FA-BSANPs和2-ME組的消除半衰期t1/2β分別為：(124.61±12.00) min和(22.17±6.00) min；兩組平均滯留時間MRT分別為(181.03±5.20) min和(22.66±2.54)min；兩組AUC分別為(290.53±10.10)(μg/ml) min和(112.99±5.50)(μg/ml) min。結果表明，2-ME-FA-BSANPs靶向給藥系統(tǒng)的消除半衰期、平均滯留時間與AUC均顯著高于2-ME組(P0.05)，顯著改善藥物2-ME半衰期短，生物利用度低等缺點，達到了提高藥物療效，降低毒副作用的目的。
[Abstract]:2 - methoxyestradiol ( 2 - methoxyestradiol , 2 - ME ) is a kind of steroid hormone compound . It is a new kind of steroid antitumor drug . It has relatively high specificity for rapidly growing tumor cells , and has no toxicity to bone marrow and hematopoietic organs .

Preparation and related properties of 1.2 - ME - FA - BSANPs .

2 - ME albumin nanoparticles ( 2 - ME - BSANPs ) were prepared by de - solvation - chemical crosslinking method . The preparation of 2 - ME - BSANPs was determined by single factor analysis of albumin concentration , 2 - ME concentration , glutaraldehyde dosage of crosslinking agent , ratio of ethanol to water , pH of solution , etc . The preparation process of 2 - ME - BSANPs was as follows : BSA concentration was 40mg / ml ;
2 - ME concentration of 4mg / ml ;
the pH value of the solution is 10.0 ;
The ratio of ethanol to water is 3 : 1 . The folic acid active ester is further used for coupling reaction with the surface active amino group of the albumin nanoparticle surface active amino group , and the folic acid - coupled 2 - ME albumin nanoparticle targeted administration system ( 2 - ME - FA - BSANPs ) is prepared so as to have an active targeting effect and improve the tumor cell uptake rate of albumin nanoparticles .

The mean particle size and potential of 2 - ME - FA - BSANPs were ( 217.5 鹵 5.1 ) nm and ( - 37.59 鹵 1.39 ) mV , respectively .
The average entrapment efficiency and the drug loading were ( 82.50 鹵 5.80 ) % and ( 7.50 鹵 0.50 ) % , respectively .
SEM images show that the particle size distribution of 2 - ME - FA - BSANPs is uniform , spherical and smooth surface .
The results of in vitro drug release showed that the drug substance 2 - ME was released rapidly , and the cumulative release was about 85 % in 12h ;
The corresponding 2 - ME - FA - BSANPs released slowly and steadily , and the cumulative release rate was about 85 % within 96 h , indicating that the targeted drug delivery system had certain sustained release characteristics .
preparing the targeted preparation into lyophilized powder , and screening to select 3 % sucrose as its freeze - drying protective agent ;
The results showed that the freeze - dried powder can be stably stored for 6 months at 4 鈩

本文編號：1962354