本文選題：氧化石墨烯 + 阿霉素　； 參考：《遼寧大學(xué)》2017年碩士論文

【摘要】：氧化石墨烯(GO)是一種新型藥物載體,GO近些年被廣泛應(yīng)用于靜注藥物的口服吸收載體,從而提高難溶性藥物的生物利用度。本文通過采用改良的新方法制備GO,并利用電荷吸引的原理在其表面進(jìn)行聚電解質(zhì)的包裹修飾。研究聚電解質(zhì)修飾GO用于抗癌藥物口服轉(zhuǎn)運(yùn)的作用及其機(jī)制,并通過電鏡表征,粒度分析,體外釋放,藥動(dòng)學(xué)研究,MTT實(shí)驗(yàn)等多方面進(jìn)行考察。本文采用改良后的Hummers法制得氧化石墨,經(jīng)超聲剝離后制得GO。采用掃描電子顯微鏡(SEM)、透射電子顯微鏡(TEM)、傅里葉紅外光譜掃描儀(FTIR)、紫外光譜(UV)等對(duì)其形貌和化學(xué)結(jié)構(gòu)進(jìn)行表征。通過表征結(jié)果證實(shí)了制備的GO是納米級(jí)的具有片層結(jié)構(gòu)的載體。相較于以前的制備GO的方法,經(jīng)我們改良之后不僅簡化了實(shí)驗(yàn)操作步驟,也獲得了安全性方面的提升。隨后研究了其對(duì)小分子量的阿霉素(DOX)和大分子量的平陽霉素(PYM)的包載和體外釋放行為。通過載藥實(shí)驗(yàn)數(shù)據(jù)分析,GO對(duì)DOX的包封率達(dá)87.58%;對(duì)PYM的包封率達(dá)到82.05%。這是由于GO結(jié)構(gòu)中含有更多的含氧官能團(tuán),所以很大程度上增加了GO的載藥能力。對(duì)GO-DOX和GO-PYM的制劑組體外釋放結(jié)果進(jìn)行分析。制劑組在不同的pH條件下表現(xiàn)出不同的釋放效果,兩組的實(shí)驗(yàn)結(jié)果又表現(xiàn)了相同的釋放趨勢,且兩組都于60h時(shí)在pH2.3的條件下達(dá)到最高的累計(jì)釋放量。說明其釋放行為表現(xiàn)出一定的pH依賴性,但是未表現(xiàn)出明顯的緩釋性。通過單因素考察法對(duì)聚丙烯酸(PAA)結(jié)合半胱氨酸(CYs)的制備工藝的最優(yōu)方法進(jìn)行了研究。通過Ellman法檢測游離巰基含量,結(jié)果表明反應(yīng)時(shí)加入的EDC含量、pH和溫度等都會(huì)影響PAA與CYs的結(jié)合,影響高分子聚合物PAA-CYs上二硫鍵的含量。因此篩選出適宜的方法,使得PAA-CYs的合成產(chǎn)率相對(duì)提高。通過電荷吸引的方法將GO-DOX和GO-PYM與巰基聚電解質(zhì)相結(jié)合并做了一系列的表征。將PAA-CYs/PAH-GO-DOX和PAA-CYs/PAH-GO-PYM進(jìn)行了透射電鏡的測定,結(jié)果呈現(xiàn)出相對(duì)穩(wěn)定的結(jié)構(gòu)。通過粒徑考察,兩種藥物的制劑組的粒徑分別為700nm與750nm左右。通過Zeta電位的考察可以看出Zeta電位為-39.8mV與-34.1mV,一方面表明出制劑的穩(wěn)定性好,另一方面也證明制劑組帶負(fù)電荷,與之前的理論分析相符合;之后又通過檢測自由巰基可看出兩種制劑組都不含有游離的巰基,可以從側(cè)面的證實(shí)PAA與CYs結(jié)合完全。最后將DOX組,GO-DOX組和PAA-CYs/PAH-GO-DOX組與PYM組、GO-PYM組和PAA-CYs/PAH-GO-PYM組分別進(jìn)行了體外釋放考察,通過數(shù)據(jù)分析可以看出,制劑組全都表現(xiàn)出了極高的耐強(qiáng)酸穩(wěn)定性,證明通過兩層聚電解質(zhì)的包裹,可使藥物在強(qiáng)酸性環(huán)境得到保護(hù);最后本課題進(jìn)行了大鼠在體腸吸收實(shí)驗(yàn)、藥動(dòng)學(xué)研究以及MTT考察。在腸吸收實(shí)驗(yàn)中,從分析結(jié)果來看,PAA-CYs/PAH-GO-PYM制劑組相比于PYM組和GO-PYM組表現(xiàn)出了更好的腸道吸收效果。主要是由于表面有PAA-CYs的包裹,使PCPGP與腸道內(nèi)壁相粘附,所以達(dá)到了更好的腸吸收效果。PCPGP制劑組在藥動(dòng)學(xué)研究方面,其AUC值較比于原料藥提高65.57mg/L·h,并且T1/2也延長了0.519h,表現(xiàn)出一定的緩釋效果。通過MTT法考察了制劑組的細(xì)胞活性研究。結(jié)果表明,在濃度20μM-100μM的條件下,PCPGP制劑組對(duì)于癌細(xì)胞A549的致死率相比于PYM組降低了2.40%。說明在24h內(nèi),PCPGP制劑組對(duì)細(xì)胞的殺傷效果沒有PYM原料藥組好。PCPGP制劑組的IC50值較之PYM組提高了24.28μM,較之GO-PYM組提高了29.42μM,說明PCPGP制劑組在相同濃度24h的時(shí)間內(nèi)表現(xiàn)出了一定的緩釋效果。最終說明PCPGP制劑組有效地提高了抗癌藥物的口服生物利用度。
[Abstract]:Graphene oxide (GO) is a new drug carrier. In recent years, GO has been widely used as an oral carrier for intravenous drug injection so as to improve the bioavailability of insoluble drugs. In this paper, a modified new method was used to prepare GO, and the polyelectrolyte package modification was carried out on its surface by the principle of charge attraction. The effect and mechanism of GO used for oral transport of anticancer drugs was investigated by electron microscopy, particle size analysis, in vitro release, pharmacokinetics and MTT experiments. The modified Hummers method was used to obtain graphite oxide, and GO. was prepared by scanning electron microscope (SEM), transmission electron microscope (TEM), and Fu after ultrasonic peeling. The morphology and chemical structure of the inner leaf infrared spectrum scanner (FTIR) and the ultraviolet spectrum (UV) were characterized. The result of characterization proved that the prepared GO was a nanoscale carrier with lamellar structure. Compared with the previous method of preparing GO, we not only simplified the procedure of the experiment, but also obtained the security. The encapsulation and in vitro release of the small molecular weight of adriamycin (DOX) and the large molecular weight of Pingyang mycin (PYM) was investigated. The encapsulation efficiency of GO to DOX was 87.58% through the drug loading experiment data, and the encapsulation efficiency of PYM was 82.05%., which was due to the presence of more oxygen functional groups in the GO structure, so the GO was greatly increased. The release results of GO-DOX and GO-PYM in vitro were analyzed. The preparation group showed different release effects under different pH conditions. The results of the two groups showed the same release trend, and the two groups all reached the highest cumulative release under the conditions of pH2.3 at the time of 60H. A certain pH dependence, but not showing obvious slow release. The optimum method of the preparation process of polyacrylic acid (PAA) binding cysteine (CYs) was studied by single factor investigation. The content of free sulfhydryl group was detected by Ellman method. The results showed that the content of EDC, pH and temperature in the reaction would affect the combination of PAA and CYs. The content of the two sulfur bond on the polymer PAA-CYs is rounded. Therefore, a suitable method is selected to make the synthesis yield of PAA-CYs relatively higher. A series of characterization of GO-DOX and GO-PYM with sulfhydryl polyelectrolytes is made by the method of charge attraction. The transmission electron microscopy of PAA-CYs/PAH-GO-DOX and PAA-CYs/PAH-GO-PYM has been measured. The results showed a relatively stable structure. The particle size of the preparation group of the two drugs was 700nm and 750nm respectively through the particle size. The Zeta potential was -39.8mV and -34.1mV through the Zeta potential investigation. On the one hand, the stability of the preparation was good, on the other hand, the negative charge of the preparation group was proved to be in conformity with the previous theoretical analysis. After the detection of free sulfhydryl group, we can see that all two kinds of preparation groups do not contain free sulfhydryl groups. It can be confirmed from the side of the combination of PAA and CYs. Finally, the release of DOX, GO-DOX and PAA-CYs/PAH-GO-DOX group with PYM group, GO-PYM group and PAA-CYs/PAH-GO-PYM group were examined respectively. All the groups showed high resistance to strong acid stability. It was proved that the drug could be protected by two layers of polyelectrolytes in the strong acid environment. Finally, the rat intestinal absorption experiment, pharmacokinetic study and MTT investigation were carried out in the rat. In the intestinal absorption experiment, the PAA-CYs/PAH-GO-PYM preparation group was compared to PYM The group and the GO-PYM group showed a better intestinal absorption effect, mainly due to the PAA-CYs parcel on the surface, which made the PCPGP adhere to the inner wall of the intestines, so it reached a better intestinal absorption effect, so the.PCPGP preparation group had a higher AUC value than the 65.57mg/ L. H compared to the API, and T1/2 also extended 0.519h. The cell activity of the preparation group was investigated by MTT method. The results showed that, under the condition of concentration of 20 mu M-100 M, the lethal rate of A549 in the PCPGP preparation group was lower than that of the PYM group. The killing effect of the PCPGP preparation group on the cells was no longer than the IC50 value of the PYM raw material group and the.PCPGP preparation group. The PYM group increased 24.28 mu M and increased 29.42 M compared with the GO-PYM group, indicating that the PCPGP preparation group showed a certain slow release effect in the time of the same concentration of 24h. Finally, the PCPGP preparation group effectively improved the oral bioavailability of the anticancer drug.

【學(xué)位授予單位】：遼寧大學(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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