【作者】 刘悦；

【导师】 郭兵；

【作者基本信息】 哈尔滨工业大学 ， 化学， 2024， 硕士

【摘要】 目前为止,脑胶质瘤依然是一种严重危害人类健康的脑重大疾病,患者面临治疗后复发率高及平均生存周期短等问题。近年来,光热疗法和免疫疗法已经成为新型癌症治疗方式,其具有安全、高效且微创等优势。目前已经开发的小分子免疫分子和光敏剂被广泛用于治疗肿瘤,但它们在生物体内表现出循环时间短,靶向效率低,生物利用度低等诸多缺点。纳米药物递送载体的出现,可以有效改善这些缺陷。值得注意的是,近年来的研究表明,以脂质体、外泌体、巨噬细胞膜、以及它们的杂化体为代表的双层膜结构纳米载体具有良好的生物相容性、可负载水溶性和油溶性药物,易于进行功能性修饰、良好的肿瘤靶向性、甚至具有免疫功能等特点,其相关研究方兴未艾。目前研究的重点问题依然是如何进一步提高这些纳米载体的药物递送效率和对疾病的治疗效果等。因此,找到适用于这些纳米载体的成像示踪分子和匹配的成像模态将对于解决这些关键问题具有重大意义。目前研究中大都使用1,1’-双十八烷基-3,3,3’,3’-四甲基吲哚菁高氯酸盐（Dil）、1,1’-双十八烷基-3,3,3,3-四甲基吲哚菁碘化物（DiR）等亲脂性荧光染料来进行活体示踪仿生纳米载体,其荧光成像大多在可见-近红外一区范围。结合最新发展的近红外二区荧光成像的优势,发展亲脂性近红外二区荧光成像探针将对仿生纳米载体的示踪具有巨大推进作用。花菁染料具有强吸收性能、较宽的发射光谱范围、荧光稳定性、生物相容性和可功能性修饰等优点,已经被广泛应用于体外和体内生物膜标记、疾病诊断和治疗等领域。在本研究中,首先设计适用于标记脂质体、外泌体和巨噬细胞膜的亲脂性近红外花菁染料,之后制备出仿生纳米药物,并评价其在近红外二区荧光成像和光热、免疫治疗脑肿瘤的效果。在论文第三部分,合成类似Dil结构的近红外花菁染料11-氯-1,1’-双十八烷基-3,3,3’,3’-四甲基-10,12-三亚甲基吲哚三碳花菁碘盐（IRC18）,以标记脂质体与基因修饰的巨噬细胞膜的杂交体,用于脑肿瘤的靶向成像和光热联合免疫治疗。具体而言,杂交体具有基因修饰的巨噬细胞膜的内源靶向性和纳米粒子的增强渗透和滞留（EPR）效应,能有效地将IRC18运送并积聚在肿瘤部位。使基因修饰的巨噬细胞膜所具有的免疫功能和IRC18的光热治疗效果相结合,使用温和光热联合免疫治疗,以起到高效治疗脑肿瘤的效果。在论文第四部分,合成具有近红外二区吸收的花菁染料NIR-C₁₂,用于标记脂质体与巨噬细胞来源的外泌体的杂交体,用于皮下脑肿瘤的靶向NIR-Ⅱ荧光成像和光热治疗。具体而言,外泌体的内源靶向性和纳米粒子的EPR效应,能有效地将NIR-C₁₂运送并积聚在肿瘤部位,从而实现NIR-Ⅱ荧光成像和光热治疗的最大化。本研究主要是合成近红外二区荧光花菁染料,研究其对脂质体-巨噬细胞膜/巨噬细胞膜的外泌体的杂化体的标记、体内示踪、及肿瘤光热/联合免疫治疗的效果。协同利用脂质体、巨噬细胞膜/巨噬细胞膜的外泌体的功能,提高对肿瘤的诊断和治疗效果。本研究中的纳米药物具有临床转化潜力。更多还原

【Abstract】 So far,glioblastoma remains a major and devastating brain disease that poses a significant threat to human health.Patients with glioblastoma often experience high rates of treatment recurrence and short average survival periods.Recently,photothermal therapy and immunotherapy have become the novel cancer treatment modality,with merits in good safety,high efficiency and minimal invasiveness.The developed small molecular immunotherapeutic drugs and photothermal agents have been widely used in cancer therapy,but they have shown shortcomings such as low circulation time,low targeting efficiency,and low bioavailability.Importantly,nanocarriers could greatly circumvent these shortcomings.Notably,recent research has demonstrated that double-layer membrane nanocarriers,such as liposomes,exosomes,macrophage membrane nanoparticles,and their hybrids,exhibit excellent biocompatibility,the ability to encapsulate both water-soluble and oil-soluble drugs,ease of functional modification,good tumor targeting properties,and e ven immunotherapeutic functionality.Studies in this field are rapidly emerging.So far,the key issue is to improve the drug delivery efficiency and therapeutic efficacy on different diseases for these nanocarriers.Importantly,to obtain highly efficient imaging tracker and corresponding imaging modalities for the nanocarriers is of high importance to solve the concern as mentioned above.Currently,lipophilic dyes such as 1,1’-bis（heptadecyl）-3,3’,3’,3’-tetramethylindocarbocyanine perchlorate（Dil）and 1,1’-bis（heptadecyl）-3,3,3’,3’-tetramethylindocarbocyanine iodide（DiR）are often used to label and trace biomimetic nanocarriers,for which the fluorescence imaging falls in visible to near-infrared imaging in the first window（NIR-I）.As fluorescence imaging in the second near-infrared window（NIR-Ⅱ）has intrinsic advantages,it is helpful to develop new lipophilic NIR-Ⅱ fluorescence probes to trace biomimetic nanocarriers.Cyanine dyes exhibit merits in strong absorbance,wide emission wavelength region,fluorescence stability,good biocompatibility,ease in modification,and have been widely used in membrane labelling,disease diagnosis,and treatment.In this study,lipophilic near-infrared cyanine dyes suitable for labeling liposomes,exosomes,and macrophage membranes were firstly designed,after which biomimetic nanomedicines were prepared and evaluated for NIR-Ⅱ fluorescence imaging and photothermal and immunotherapeutic treatment of brain tumors.In the third part of the thesis,the NIR cyanine dyes 11-chloro-1,1’-bis（heptadecyl）-3,3,3’,3’-tetramethyl-10,12-trimethyleneindotricarbocyanine iodide（IRC18）with similar structure to Dil is formulated for labelling hybrid of liposomes and gene-edited macrophage membrane for targeted imaging and combinatory photothermal and immunotherapy of brain tumors.In details,the hybrid nanomedicine has intrinsic targeting capability from gene-edited macrophage membrane,and enhanced penetration and enhanced permeability and retention（EPR）effect,could efficiently deliver IRC18 to tumor tissue.With combining the immunotherapeutic functions of macrophage membrane and photothermal effect of IRC18,the nanomedicine could execute high efficacy in tumor killing under mild photothermal conditions.In the fourth part of the thesis,the cyanine dye NIR-C₁₂ with NIR-Ⅱ absorbance is synthesized to label hybrid constructs of liposomes and macrophage-derived exosomes,which is further used for targeted imaging and photothermal therapy of brain tumors.Specifically,the intrinsic targeting of exosomes and EPR effect of nanoparticles enable effective transportation and accumulation of NIR-C₁₂ at the tumor site,thereby achieving maximized NIR-Ⅱ fluorescence imaging and photothermal therapy.The main objective of this study is to synthesize NIR fluorescent cyanine dye and investigate its labeling,in vivo tracking,and effects on hybrid constructs of liposome-macrophage membrane/exosome.The study aims to evaluate the therapeutic efficacy of tumor photothermal therapy and combined immunotherapy.By leveraging the functionalities of liposomes and macrophage membrane/exosomes,this research aims to enhance the diagnosis and treatment of tumors.The nano-drug developed in this study holds potential for clinical translation.更多还原