本文選題：達托霉素 + 長循環(huán)脂質(zhì)體　； 參考：《西南大學(xué)》2014年碩士論文

【摘要】：隨著病原體耐藥防控難度的持續(xù)增大,除不斷研發(fā)新型抗生素之外,對已有抗生素進行遞送研究與新制劑創(chuàng)制具有同等重要性。達托霉素是近年來上市的-種新型脂肽類抗生素,目前僅有其普通溶液型注射劑用于臨床。本文采用聚乙二醇修飾的脂質(zhì)體作為載體,考察并系統(tǒng)評價經(jīng)優(yōu)化制備的長循環(huán)達托霉素脂質(zhì)體(PLD)抗耐甲氧西林金葡菌(MRSA252)全身感染。通過優(yōu)化制備得到的達托霉素脂質(zhì)體粒徑(111.5±15.4)nm、載藥量較高(5.81±0.19)%,并具有良好穩(wěn)定性。體內(nèi)外評價結(jié)果顯示,在相同劑量下PLD與達托霉素原料藥及普通脂質(zhì)體制劑相比,對MRSA252有更持久的抑制效果,并能顯著提高耐甲氧西林金葡菌全身感染模型小鼠的存活率。此外,PLD能顯著提高達托霉素在肺部的分布并顯示良好的安全性。上述研究結(jié)果提示,長循環(huán)脂質(zhì)化達托霉素對抗MRSA嚴重感染(血源性肺炎等)具有良好的應(yīng)用前景,可為達托霉素新制劑的研發(fā)及臨床相關(guān)研究提供重要參考。 本課題主要研究內(nèi)容如下：(1)達托霉素長循分析方法的建立 確定達托霉素的最大吸收波長為221nm,并且還確定了HPLC檢測達托霉素的色譜條件： 色譜柱：Inertsil ODS-SP C18柱(150mm×5mm,5μm) 流動相：乙腈-0.5%磷酸二氫銨緩沖液=40：60 流速：0.7mL/min;檢測波長：221nm進樣量：20μ L；柱溫：30℃出峰時間：6.5±0.2min最低檢測限：0.60ng還通過回收率實驗、精密度實驗得出此種檢測方法準確且重現(xiàn)性好。 (2)達托霉素長循環(huán)脂質(zhì)體的制備及相關(guān)性質(zhì)考察 通過考察制備達托霉素長循環(huán)脂質(zhì)體的幾個影響因素：氫化卵磷脂(HSPC)與膽固醇的摩爾比、脂藥質(zhì)量比以及高壓均質(zhì)機循環(huán)次數(shù)。最終得出制備達托霉素長循環(huán)脂質(zhì)體的最優(yōu)處方為,氫化卵磷脂：膽固醇：mPEG2ooo-DSPE=18：6：1(摩爾比),脂藥比=15：1(質(zhì)量比),水化液經(jīng)高壓均質(zhì)機1000bars循環(huán)均質(zhì)10次,就能得到穩(wěn)定的達托霉素長循環(huán)脂質(zhì)體。由最優(yōu)制備工藝所制得的脂質(zhì)體,粒徑為(111.5±15.4)nm, Zeta電位為(-15.1±2.8)mV,載藥量為(5.81±0.19)%,包封率為(92.56±3.15)%,并且性質(zhì)穩(wěn)定,在4-C的冰箱中保存2個月后幾乎無泄漏、無沉淀現(xiàn)象發(fā)生。 (3)體外抗耐甲氧西林金葡菌實驗 主要考察達托霉素三個制劑：達托霉素原料藥水溶液DS、普通達托霉素脂質(zhì)體CLD、達托霉素長循環(huán)脂質(zhì)體PLD對MRSA252生長的影響,結(jié)果表明,PLD對MRSA252的抑制效果更持久。 (4)藥動學(xué)及組織分布研究 結(jié)果表面達托霉素長循環(huán)脂質(zhì)體延長了達托霉素在大鼠血液里的循環(huán)時間,且明顯的增加了達托霉素在肝、脾、肺的分布量。 (5)體內(nèi)藥效學(xué)、安全性評價 通過比較DS、CLD、PLD對MRSA252致全身感染KM小鼠的治療效果比較,結(jié)果顯示,尾靜脈注射單次次給藥達托霉素長循環(huán)脂質(zhì)體就能使感染KM小鼠的存活率在觀察期(7天)提高到80%。安全性評價結(jié)果表明達托霉素長循環(huán)脂質(zhì)體未對KM小鼠有明顯毒性。
[Abstract]:With the increasing difficulty of drug resistance and prevention and control, in addition to the continuous development of new antibiotics, the research on the delivery of the existing antibiotics is of equal importance to the creation of new preparations. DDT is a new type of lipopeptide antibiotic in recent years. At present, only the common solution injection is used in clinical. This paper uses poly (two) The alcohol modified liposome was used as a carrier to evaluate and systematically evaluate the optimized long circulation dacostin liposome (PLD) against methicillin resistant Staphylococcus aureus (MRSA252) systemic infection. Through the optimization of the size of the liposomes (111.5 + 15.4) nm, the drug loading is higher (5.81 + 0.19)%, and has good stability. The results showed that, at the same dose, PLD had a more persistent inhibitory effect on MRSA252 compared with the common liposomal drug and the common liposome agent, and could significantly increase the survival rate of the model mice with methicillin-resistant Staphylococcus aureus systemic infection. In addition, PLD could significantly improve the distribution of DTM in the lung and show good safety. The results suggest that long circulatory liposomes have good application prospects against MRSA severe infection (hematogenous pneumonia, etc.), and can provide an important reference for the research and development of the new preparation of dalamycin and clinical related research.
The main contents of this research are as follows: (1) establishment of DAC analysis method
The maximum absorption wavelength of Das was determined to be 221nm, and the chromatographic conditions for HPLC detection of Das were also determined.
Chromatographic column: Inertsil ODS-SP C18 column (150mm x 5mm, 5 m)
Mobile phase: acetonitrile -0.5% phosphoric acid two hydrogen ammonium buffer =40:60
Flow rate: 0.7mL/min; detection wavelength: 221nm sampling amount: 20 mu L; column temperature: 30 C peak time: 6.5 + 0.2min minimum detection limit: 0.60ng is also through recovery experiment, precision experiment results show that this method is accurate and reproducible.
(2) preparation and related properties of long circulating liposomes of DTaP
Several factors affecting the preparation of the long circulating liposomes of dalochomycin: the molar ratio of HSPC to cholesterol, the mass ratio of lipoid drugs and the cycles of high pressure homogenizer. The optimal formulation for the preparation of the long circulating liposome of dalotamiin was obtained, and the hydrogenated oophospholipid: cholesterol: mPEG2ooo-DSPE=18:6:1 (mole ratio) When the liposomes are compared with the =15:1 (mass ratio), the hydrated liquid is homogenized 10 times through the high pressure homogenizer 1000bars, and the stable dapycin long circulating liposome can be obtained. The liposomes obtained by the optimal preparation process are (111.5 + 15.4) nm, the Zeta potential is (-15.1 2.8) mV, the drug loading is (5.81 + 0.19)%, the encapsulation rate is (92.56 + 3.15)%, and the property is stable. It has been stored in the refrigerator of 4-C for 2 months, and there is almost no leakage.
(3) the experiment of anti methicillin resistant Staphylococcus aureus in vitro
Three preparations of dalotomiin were mainly studied: DS, CLD of dalotomiin liposome, and the effect of PLD on the growth of MRSA252. The results showed that the inhibition effect of PLD to MRSA252 was more lasting.
(4) study on pharmacokinetics and tissue distribution
Results DDT lengthened the circulation time of dalotamicin in the blood of rats, and significantly increased the distribution of dalamycin in the liver, spleen and lung.
(5) in vivo pharmacodynamics, safety evaluation
By comparing the therapeutic effects of DS, CLD, and PLD on KM mice induced by MRSA252, the results showed that the single dose of tataycin long circulating liposome injected into the tail vein could increase the survival rate of the infected KM mice in the observation period (7 days) to the 80%. security evaluation results, indicating that the long circulating liposomes of tataycin did not have obvious toxicity to KM mice. Sex.

【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R978.1

【參考文獻】

相關(guān)期刊論文 前8條

1 穆筱梅;梁世強;;乳化法制備聚乙烯醇修飾的果酸脂質(zhì)體[J];日用化學(xué)工業(yè);2006年06期

2 吳燕;吳誠;梅興國;呂萬良;;鹽酸表柔比星長循環(huán)熱敏凍干脂質(zhì)體的處方工藝研究與體外釋藥機制探討[J];軍事醫(yī)學(xué)科學(xué)院院刊;2010年02期

3 程志祥,徐建民;一種新的抗革蘭陽性菌制劑達托霉素[J];中國新藥與臨床雜志;2001年06期

4 曾昭武;王小麗;張陽德;;替加氟磁性長循環(huán)脂質(zhì)體在大鼠體內(nèi)的藥代動力學(xué)和靶向性[J];中國藥理學(xué)與毒理學(xué)雜志;2009年01期

5 王世霞;曹德英;;聚合物修飾長循環(huán)脂質(zhì)體研究進展[J];中國藥房;2008年34期

6 儲茂泉,古宏晨,劉國杰;丹參酮脂質(zhì)體的藥物滲漏和微粒聚結(jié)特性研究[J];中國醫(yī)藥工業(yè)雜志;2002年08期

7 顏仁梁;劉志剛;;黃芩素長循環(huán)脂質(zhì)體處方優(yōu)化研究[J];中藥材;2010年01期

8 王軍;;殼聚糖包衣的維生素E脂質(zhì)體的制備及其體外性質(zhì)考察[J];中國藥師;2008年07期

，

本文編號：1824467