Her2抗體介導(dǎo)的藥物傳遞與靶向治療
發(fā)布時(shí)間：2018-11-09 16:27
【摘要】：多年來,癌癥一直對(duì)人類的健康構(gòu)成巨大威脅。目前治療癌癥最常用的方法是化療。但化療存在很大的弊端,由于化療藥物在體內(nèi)不能特異性識(shí)別腫瘤,在殺死腫瘤細(xì)胞的同時(shí),也會(huì)殺傷正常組織,從而帶來嚴(yán)重的毒副作用。光動(dòng)力療法(Photodynamic Therapy, PDT)是近年來興起的一種治療癌癥的新方法。其中光敏劑是PDT的關(guān)鍵物質(zhì)。然而絕大多數(shù)光敏劑都是不溶于水,或在生理?xiàng)l件下容易聚合。這大大限制PDT的發(fā)展。本實(shí)驗(yàn)以疏水性吲哚菁綠(ICG-ODA)為光敏劑,阿霉素(doxorubicin, DOX)為化療藥,以Her2抗體為靶頭,采用硫酸銨梯度法制備了一種具有靶向特異性的光敏脂質(zhì)體。本實(shí)驗(yàn)采用單因素法篩選處方,制備出了粒徑為128.1±1.8nm的光敏脂質(zhì)體(PSL),粒徑大小均一,穩(wěn)定性高。脂質(zhì)體的結(jié)構(gòu)清晰,形態(tài)良好。DOX包封率高達(dá)90%以上。對(duì)測(cè)定包封率的影響因素考察結(jié)果表明包封率的測(cè)定方法科學(xué),結(jié)果準(zhǔn)確、可靠。光敏脂質(zhì)體的體外釋放實(shí)驗(yàn)中,經(jīng)近紅外激光(NIR)照射后,DOX會(huì)快速且持續(xù)地從光敏脂質(zhì)體中釋放出來。結(jié)果顯示在近紅外激光照射條件下,6h內(nèi)DOX的累積釋放量比無近紅外激光照射時(shí)提高了近20%,近紅外激光觸發(fā)DOX從脂質(zhì)體釋放的效果明顯。在以上光敏脂質(zhì)體的基礎(chǔ)上,我們又以DSC為中間嫁接物,制備出了PEG化修飾的Her2抗體,再通過與光敏脂質(zhì)體進(jìn)行共孵育制備得到Her2靶向的光敏脂質(zhì)體(Her2-PSL)。該靶向脂質(zhì)體可以特異性地被MCF-7攝取。在MCF-7和A549細(xì)胞攝取差異實(shí)驗(yàn)中,MCF-7細(xì)胞中的熒光信號(hào)強(qiáng)度遠(yuǎn)高于A549細(xì)胞,表明Her2-PSL能夠特異性地主動(dòng)靶向至MCF-7細(xì)胞。荷MCF-7和荷A549腫瘤的動(dòng)物模型體內(nèi)分布研究顯示,MCF-7腫瘤內(nèi)熒光信號(hào)明顯也強(qiáng)于A549腫瘤,表明攜帶Her2抗體的光敏脂質(zhì)體能夠更多地靶向至MCF-7實(shí)體瘤。在抗腫瘤藥效學(xué)研究中,結(jié)果顯示照射近紅外激光后,治療組腫瘤抑制率為93.6%,且體重曲線變化波動(dòng)很小,展現(xiàn)出顯著的抗腫瘤效果。實(shí)驗(yàn)結(jié)果表明,該光敏脂質(zhì)體與Her2抗體結(jié)合后可以選擇性靶向到MCF-7腫瘤部位,并在給予近紅外激光照射后釋放出大量DOX,實(shí)現(xiàn)化療與PDT的協(xié)同治療,療效顯著。二者協(xié)同治療在抗腫瘤治療領(lǐng)域有著光明的前景。
[Abstract]:Cancer has been a great threat to human health for many years. The most commonly used treatment for cancer is chemotherapy. However, chemotherapeutic drugs have great disadvantages. Because chemotherapy drugs can not specifically recognize tumor in vivo, it can kill tumor cells and kill normal tissues, which brings serious toxic side effects. Photodynamic therapy (Photodynamic Therapy, PDT) is a new method to treat cancer in recent years. Guang Min is the key substance of PDT. However, most Guang Min agents are insoluble in water or easily polymerized under physiological conditions. This greatly limits the development of PDT. Using hydrophobic indocyanine green (ICG-ODA) as Guang Min, doxorubicin (doxorubicin, DOX) as chemotherapeutic agent, and Her2 antibody as target, a kind of liposome with targeted specificity was prepared by ammonium sulfate gradient method. In this experiment, the single factor method was used to screen the prescription, and the liposome (PSL), with a diameter of 128.1 鹵1.8nm was prepared with uniform particle size and high stability. The structure and morphology of liposomes were clear and the entrapment efficiency of DOX was over 90%. The results of investigation on the factors influencing the determination of encapsulation efficiency show that the determination method of encapsulation efficiency is scientific, and the results are accurate and reliable. In the in vitro release of Guang Min liposomes, DOX was released rapidly and continuously from Guang Min liposomes after near-infrared laser (NIR) irradiation. The results showed that under the condition of NIR laser irradiation, the cumulative release amount of DOX in 6 h was increased by nearly 20% than that without NIR laser irradiation, and the effect of NIR laser triggering DOX release from liposome was obvious. On the basis of the above Guang Min liposomes, we prepared the Her2 antibody modified by PEG with DSC as the intermediate grafted material. Then we co-incubated with Guang Min liposomes to obtain the Her2 targeted Guang Min liposomes (Her2-PSL). The targeted liposome can be specifically ingested by MCF-7. The fluorescence intensity of MCF-7 cells was much higher than that of A549 cells in the uptake difference assay between MCF-7 and A549 cells, which indicated that Her2-PSL could target MCF-7 cells specifically and actively. The distribution of MCF-7 and A549 tumor models in vivo showed that the fluorescence signal in MCF-7 tumor was stronger than that in A549 tumor, indicating that Guang Min liposome carrying Her2 antibody could target more MCF-7 solid tumor. In the study of antitumor pharmacodynamics, the results showed that the tumor inhibition rate of the treatment group was 93.6 after NIR laser irradiation, and the change of body weight curve was very small, showing significant antitumor effect. The results showed that the liposome combined with Her2 antibody could selectively target to the tumor site of MCF-7 and release a large amount of DOX, after irradiation with NIR laser to achieve the synergistic effect of chemotherapy and PDT. The synergistic therapy has a bright prospect in the field of anti-tumor therapy.
【學(xué)位授予單位】：哈爾濱商業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R943
[Abstract]:Cancer has been a great threat to human health for many years. The most commonly used treatment for cancer is chemotherapy. However, chemotherapeutic drugs have great disadvantages. Because chemotherapy drugs can not specifically recognize tumor in vivo, it can kill tumor cells and kill normal tissues, which brings serious toxic side effects. Photodynamic therapy (Photodynamic Therapy, PDT) is a new method to treat cancer in recent years. Guang Min is the key substance of PDT. However, most Guang Min agents are insoluble in water or easily polymerized under physiological conditions. This greatly limits the development of PDT. Using hydrophobic indocyanine green (ICG-ODA) as Guang Min, doxorubicin (doxorubicin, DOX) as chemotherapeutic agent, and Her2 antibody as target, a kind of liposome with targeted specificity was prepared by ammonium sulfate gradient method. In this experiment, the single factor method was used to screen the prescription, and the liposome (PSL), with a diameter of 128.1 鹵1.8nm was prepared with uniform particle size and high stability. The structure and morphology of liposomes were clear and the entrapment efficiency of DOX was over 90%. The results of investigation on the factors influencing the determination of encapsulation efficiency show that the determination method of encapsulation efficiency is scientific, and the results are accurate and reliable. In the in vitro release of Guang Min liposomes, DOX was released rapidly and continuously from Guang Min liposomes after near-infrared laser (NIR) irradiation. The results showed that under the condition of NIR laser irradiation, the cumulative release amount of DOX in 6 h was increased by nearly 20% than that without NIR laser irradiation, and the effect of NIR laser triggering DOX release from liposome was obvious. On the basis of the above Guang Min liposomes, we prepared the Her2 antibody modified by PEG with DSC as the intermediate grafted material. Then we co-incubated with Guang Min liposomes to obtain the Her2 targeted Guang Min liposomes (Her2-PSL). The targeted liposome can be specifically ingested by MCF-7. The fluorescence intensity of MCF-7 cells was much higher than that of A549 cells in the uptake difference assay between MCF-7 and A549 cells, which indicated that Her2-PSL could target MCF-7 cells specifically and actively. The distribution of MCF-7 and A549 tumor models in vivo showed that the fluorescence signal in MCF-7 tumor was stronger than that in A549 tumor, indicating that Guang Min liposome carrying Her2 antibody could target more MCF-7 solid tumor. In the study of antitumor pharmacodynamics, the results showed that the tumor inhibition rate of the treatment group was 93.6 after NIR laser irradiation, and the change of body weight curve was very small, showing significant antitumor effect. The results showed that the liposome combined with Her2 antibody could selectively target to the tumor site of MCF-7 and release a large amount of DOX, after irradiation with NIR laser to achieve the synergistic effect of chemotherapy and PDT. The synergistic therapy has a bright prospect in the field of anti-tumor therapy.
【學(xué)位授予單位】：哈爾濱商業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R943
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