腫瘤靶向D構(gòu)型多肽納米纖維作為10-羥基喜樹堿載體的研究
發(fā)布時(shí)間：2018-09-04 13:58
【摘要】：自組裝納米藥物載體在藥物控制釋放及靶向藥物傳輸方面,已經(jīng)發(fā)揮了巨大地推動(dòng)作用。研究證實(shí),除化學(xué)組成、尺寸大小及表面電位等因素外,納米載體的形狀對(duì)其在體內(nèi)的代謝也具有重要影響。對(duì)于某些載體材料,納米纖維比納米微球表現(xiàn)出更少的網(wǎng)狀內(nèi)皮系統(tǒng)攝取和更強(qiáng)的腫瘤組織選擇性。多肽納米材料因生物相容性好,易連接功能基團(tuán),且可以根據(jù)需要“自下而上”地設(shè)計(jì)序列而被廣泛應(yīng)用。自組裝多肽納米纖維在藥物傳輸領(lǐng)域具有很大的應(yīng)用前景。 由天然氨基酸(L氨基酸)組成的納米纖維在體內(nèi)容易被各種蛋白酶催化降解,存在半衰期短、穩(wěn)定性差等缺點(diǎn),可能導(dǎo)致負(fù)載藥物的過(guò)早釋放,在一定程度上限制了其作為藥物載體在體內(nèi)的應(yīng)用。將D構(gòu)型的非天然氨基酸引入到多肽序列中,能夠降低體內(nèi)各種消化蛋白酶的識(shí)別和降解,比L構(gòu)型的氨基酸表現(xiàn)出更高的穩(wěn)定性。精氨酸-甘氨酸-天冬氨酸(arginine-glycine-aspartic, RGD)三肽模型是整合素αvβ3的結(jié)合基序,而整合素αvβ3過(guò)度表達(dá)于惡性腫瘤內(nèi)皮細(xì)胞,對(duì)癌細(xì)胞的粘附、信號(hào)轉(zhuǎn)導(dǎo)、遷移和生存至關(guān)重要,被廣泛應(yīng)用于腫瘤的診斷和靶向治療。因此,我們?cè)O(shè)計(jì)并合成了具有腫瘤靶向性的由L構(gòu)型氨基酸構(gòu)成的多肽分子Nap-GFFYGRGD(用L-peptide表示)和由D構(gòu)型氨基酸構(gòu)成的多肽分子Nap-GDFDFDYGRGD(用D-peptide表示),將多肽分子自組裝形成的納米纖維作為疏水性抗腫瘤藥物10-羥基喜樹堿(10-hydroxycamptothecin, HCPT)的載體用于體內(nèi)外腫瘤治療的研究。實(shí)驗(yàn)結(jié)果表明：L-peptide和D-peptide經(jīng)加熱煮沸自然冷卻后可自組裝形成直徑為10-20nm的納米纖維(分別用L-fiber和D-fiber表示)；與L-fiber相比,D-fiber具有更強(qiáng)地抵抗蛋白酶K降解的能力,說(shuō)明D氨基酸的引入提高了多肽納米纖維的穩(wěn)定性,為其體內(nèi)應(yīng)用奠定了基礎(chǔ)；通過(guò)氯胺-T法對(duì)多肽進(jìn)行放射性同位素125Ⅰ標(biāo)記,體內(nèi)分布實(shí)驗(yàn)結(jié)果表明L-peptide和D-peptide具有不同的體內(nèi)代謝規(guī)律,L-fiber主要分布在胃中,而D-fiber主要分布于肝臟,二者在腫瘤部位均具有明顯的積累；L-peptide和D-peptide可通過(guò)疏水作用與HCPT共組裝形成可注射的載藥納米纖維(分別用L-fiber-HCPT和D-fiber-HCPT表示),實(shí)驗(yàn)表明以L-fiber和D-fiber為載體可以提高HCPT在生理環(huán)境中的溶解度,分別約51倍和46倍,并且在細(xì)胞和體內(nèi)水平上提高了HCPT的抗腫瘤效果。因此,D-fiber-HCPT與L-fiber-HCPT相比具有更好的體外穩(wěn)定性和體內(nèi)腫瘤治療效果。 通過(guò)本論文我們完成了一種能夠?qū)κ杷钥鼓[瘤藥物HCPT增溶、提高其體內(nèi)外抗腫瘤效果并降低毒副作用的D構(gòu)型多肽納米纖維載體的研究,為腫瘤靶向的治療及多肽藥物載體的設(shè)計(jì)提供新的思路和方法。
[Abstract]:Self-assembled nano-drug carriers have played an important role in drug delivery and drug delivery. In addition to chemical composition, size and surface potential, the shape of nano-carrier also has an important effect on its metabolism in vivo. For some carrier materials, nanofibers exhibit less reticuloendothelial uptake and stronger tumor tissue selectivity than nano-microspheres. Polypeptide nanomaterials are widely used because of their good biocompatibility, easy linking of functional groups and "bottom-up" design of sequences according to their needs. Self-assembled polypeptide nanofibers have great application prospect in the field of drug transport. Nanofibers composed of natural amino acids (L-amino acids) are easily degraded by various proteases in vivo, which may lead to the premature release of loaded drugs due to their short half-life and poor stability. To some extent, it limits its application as drug carrier in vivo. The introduction of non-natural D-shaped amino acids into polypeptide sequences can reduce the recognition and degradation of various digestive proteases in the body and exhibit higher stability than L-configuration amino acids. Arginine glycine aspartic acid (arginine-glycine-aspartic, RGD) tripeptide model is the binding motif of integrin 偽 v 尾 3, and integrin 偽 v 尾 3 overexpression in malignant tumor endothelial cells, adhesion, signal transduction, migration and survival of cancer cells is very important. It is widely used in tumor diagnosis and targeted therapy. therefore We have designed and synthesized polypeptide molecule Nap-GFFYGRGD (expressed by L-peptide) and polypeptide molecule Nap-GDFDFDYGRGD (expressed by D-peptide), which is composed of L-configuration amino acid and D-configuration amino acid, which has tumor targeting property. Fiber was used as a carrier of 10-hydroxycamptothecin (HCPT) as a hydrophobic antitumor drug in vitro and in vivo. The experimental results show that: L-peptide and D-peptide can self-assemble into nanofibers with diameter of 10-20nm (expressed as L-fiber and D-fiber, respectively) after heated and boiled naturally cooled, and D-fiber has stronger ability to resist protease K degradation than L-fiber. It shows that the introduction of D-amino acid improves the stability of polypeptide nanofibers and lays the foundation for its application in vivo. The results of in vivo distribution experiment showed that L-peptide and D-peptide had different metabolic laws in the stomach, while D-fiber mainly distributed in the liver, and both had obvious accumulation in the tumor site. L-peptide and D-peptide can co-assemble with HCPT to form injectable drug-loaded nanofibers (expressed by L-fiber-HCPT and D-fiber-HCPT respectively) through hydrophobic interaction. The experimental results show that L-fiber and D-fiber can improve the solubility of HCPT in physiological environment, about 51 times and 46 times, respectively. It also improves the anti-tumor effect of HCPT at cell and in vivo level. Therefore, D-fiber-HCPT has better in vitro stability and in vivo tumor therapy than L-fiber-HCPT. In this paper, we have completed a novel D-configuration polypeptide nanofiber carrier which can solubilize hydrophobic antitumor drug HCPT, improve its antitumor effect in vivo and in vitro, and reduce toxic side effects. To provide new ideas and methods for tumor targeting therapy and the design of polypeptide drug carrier.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R943
本文編號(hào)：2222310
[Abstract]:Self-assembled nano-drug carriers have played an important role in drug delivery and drug delivery. In addition to chemical composition, size and surface potential, the shape of nano-carrier also has an important effect on its metabolism in vivo. For some carrier materials, nanofibers exhibit less reticuloendothelial uptake and stronger tumor tissue selectivity than nano-microspheres. Polypeptide nanomaterials are widely used because of their good biocompatibility, easy linking of functional groups and "bottom-up" design of sequences according to their needs. Self-assembled polypeptide nanofibers have great application prospect in the field of drug transport. Nanofibers composed of natural amino acids (L-amino acids) are easily degraded by various proteases in vivo, which may lead to the premature release of loaded drugs due to their short half-life and poor stability. To some extent, it limits its application as drug carrier in vivo. The introduction of non-natural D-shaped amino acids into polypeptide sequences can reduce the recognition and degradation of various digestive proteases in the body and exhibit higher stability than L-configuration amino acids. Arginine glycine aspartic acid (arginine-glycine-aspartic, RGD) tripeptide model is the binding motif of integrin 偽 v 尾 3, and integrin 偽 v 尾 3 overexpression in malignant tumor endothelial cells, adhesion, signal transduction, migration and survival of cancer cells is very important. It is widely used in tumor diagnosis and targeted therapy. therefore We have designed and synthesized polypeptide molecule Nap-GFFYGRGD (expressed by L-peptide) and polypeptide molecule Nap-GDFDFDYGRGD (expressed by D-peptide), which is composed of L-configuration amino acid and D-configuration amino acid, which has tumor targeting property. Fiber was used as a carrier of 10-hydroxycamptothecin (HCPT) as a hydrophobic antitumor drug in vitro and in vivo. The experimental results show that: L-peptide and D-peptide can self-assemble into nanofibers with diameter of 10-20nm (expressed as L-fiber and D-fiber, respectively) after heated and boiled naturally cooled, and D-fiber has stronger ability to resist protease K degradation than L-fiber. It shows that the introduction of D-amino acid improves the stability of polypeptide nanofibers and lays the foundation for its application in vivo. The results of in vivo distribution experiment showed that L-peptide and D-peptide had different metabolic laws in the stomach, while D-fiber mainly distributed in the liver, and both had obvious accumulation in the tumor site. L-peptide and D-peptide can co-assemble with HCPT to form injectable drug-loaded nanofibers (expressed by L-fiber-HCPT and D-fiber-HCPT respectively) through hydrophobic interaction. The experimental results show that L-fiber and D-fiber can improve the solubility of HCPT in physiological environment, about 51 times and 46 times, respectively. It also improves the anti-tumor effect of HCPT at cell and in vivo level. Therefore, D-fiber-HCPT has better in vitro stability and in vivo tumor therapy than L-fiber-HCPT. In this paper, we have completed a novel D-configuration polypeptide nanofiber carrier which can solubilize hydrophobic antitumor drug HCPT, improve its antitumor effect in vivo and in vitro, and reduce toxic side effects. To provide new ideas and methods for tumor targeting therapy and the design of polypeptide drug carrier.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R943
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 楊柯君;;全球癌癥狀況最新數(shù)據(jù)更新[J];上海醫(yī)藥;2014年02期
，本文編號(hào)：2222310
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