兰州大学机构库 >化学化工学院
Alternative TitleDesign, synthesis and biological application of Baylis–Hillman derivatives as probes and potential drugs
Thesis Advisor房建国
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name理学博士
Degree Discipline化学
Keyword生物巯基 Baylis–Hillman衍生物 荧光探针 药物控释系统 Trx系统抑制剂
Abstract生物巯基在生命系统中具有重要的作用,其含量或氧化还原水平的变化与许多疾病密切相关。因此,发展高效直观的方法对生物巯基进行检测或以巯基为靶点进行药物设计显得十分重要。本论文基于Baylis-Hillman(BH)衍生物与亲核性巯基可发生加成-消除-再加成的反应性质,合成了一系列BH衍生结构的荧光探针和药物分子,分别用于小分子生物巯基的检测、蛋白中邻二巯基的检测、巯基为靶点的荧光前药设计、硫氧还蛋白系统抑制剂的设计中。BH衍生结构的荧光探针或药物设计为进一步构建性能优异的探针、新药或药物控释体系奠定了基础。主要内容如下: 1. 概述了特异性检测小分子生物巯基和蛋白邻近巯基荧光探针的研究进展。接着对近年来可视化荧光前药体系的发展情况进行综述。最后,对BH类衍生物在药物设计和蛋白修饰中的应用进行了归纳总结。 2. 基于BH衍生物具有巯基点击-释放的特点,设计合成了特异性识别生物巯基的BH类探针分子MHA-green。测试表明,该探针能够快速、选择性识别生物巯基,并能应用于细胞和斑马鱼活体中进行生物巯基的荧光成像。这一探针的设计应用为进一步发展基于BH衍生结构的探针或药物提供了理论基础。 3. 邻二巯基蛋白(VDPs)在维持细胞内氧化还原平衡方面发挥重要作用,而VDPs水平的异常与许多疾病密切相关。因此,发展特异性检测VDPs的方法是十分重要的。基于BH衍生物与硫醇双亲核加成的特点,我们将BH衍生化的丙烯酰基连接到二乙氨基香豆素荧光团上,合成了6个BH类VDPs探针。筛选发现探针AC-green对VDPs具有快速、特异性的响应,并可进一步应用于细胞和斑马鱼活体中进行VDPs的检测成像。利用了探针对帕金森疾病模型中还原态VDPs水平的变化进行监测,首次揭示了该类疾病与蛋白邻近巯基的氧化有一定关联,为疾病的诊断治疗提供一定的参考。AC-green的设计合成为进一步的发展检测VDPs的小分子探针提供了新的策略。 4. 可控释放策略在探针和前药设计方面受到越来越多的关注。基于BH衍生物与巯基点击-释放的特点,我们将BH加成物作为药物递送系统的模块分子,构建了一个具有广泛兼容性的AH类荧光前药体系,实现对不同官能团分子(如胺基、羟基、羧基、巯基等)的可视化激活释放。同时,利用AH类荧光前药双光子成像的优势,连接喜树碱和一氧化氮供体的前药AH-CPT和AH-NO可以在细胞、斑马鱼活体和小鼠大脑组织中激活并可视化释放,AH-NO可控释放NO的特点也使其在中风模型中具有保护神经细胞的作用。基于BH加成物的荧光前药体系,具有易于制备、官能团广泛兼容性、响应速度快、释放效率高等优点,有望在药物控释系统的构建上得到广泛的应用。 5. 基于酰胺类BH衍生物对邻二巯基蛋白有更好反应选择性的特点,我们采用Ugi反应,设计合成了一系列的BH类拟肽分子,通过细胞毒活性的筛选,得到了活性较好的化合物5f,其可以选择性抑制HepG2细胞的生长。通过对其作用机制的研究,发现化合物5f可通过抑制硫氧还蛋白系统来发挥抑制肿瘤生长的作用。这一研究为进一步开发BH类药物分子提供了理论基础。
Other AbstractBiological thiols play vital roles in many biological processes. Abnormal change of thiols level or redox are associated with many diseases. Therefore, it is very important to develop efficient methods to detect biological thiols or to design drugs with thiols as targets. In this thesis, we reported herein a new strategy for constructing fluorescent probes, fluorogenic controlled release system and inhibitors by employing the specific reaction between thiols and BH derivatives. The structural novelty of probes or drugs would provide guidance for developing novel probes, controlled release system and potential drugs. Details of this dissertation were summarized as follows: 1. A brief review of the fluorescent probes for small molecule thiols and VDPs were presented. Then, we summarized the application of BH derivatives in drug design and protein modification. In addition, the development of fluorogenic release system in recent years were also reviewed. 2. Conjugation of the BH derivative (methyl 2-(hydroxymethyl)acrylate) with naphthalimide via a carbamate linker generates the fluorescent probe MHA-green. The probe displays low toxicity, excellent sensitivity towards thiols. The probe was also successfully applied to detect biological thiols in cells and zebrafish. 3. Vicinal dithiol-containing proteins (VDPs) play vital roles in regulating the cellular redox balance. Abnormal change VDPs level is associated with many diseases. We describd in this manuscript a structurally-novel β-allyl carbamate fluorescent probe (AC-green) based on the Baylis-Hillman reaction. The probe exhibits low toxicity, and features high selectivity and rapid response towards VDPs. The sensing mechanism of the probe was clarified and the probe is ready to image VDPs in living cells and zebrafishes. We further demonstrated the application of the probe to trace global changes of VDPs level in Parkinsonism, linking the decrease of VDPs to Parkinson’s disease for the first time. In addition, the success of the probe also provides a new scaffold to advance constructing VDPs probes. 4. The controlled release of a molecule of interest (MOI) has gained increasing interests as this strategy is particularly useful in probe design, prodrug construction and drug delivery. We report herein a versatile thiol-triggered fluorogenic release system using the Baylis-Hillman (BH) adducts as a modular template. Various common functional groups (e.g. amino group, hydroxyl group, carboxylic group and sulfhydryl group) in a MOI can be readily integrated into the BH adduct, demonstrating a broad scope of functional group compatibility. We further prepared two prodrugs to release camptothecin and nitric oxide (NO) using the BH adduct template that containing a two-photon excitable fluorophore. By taking the advantage of the two-photon imaging technology, the activation of prodrugs was revealed in live cells and in mouse brain tissues. The therapeutic potential of the NO prodrug was also validated in a cellular model of stroke. This BH adduct-based fluorogenic controlled release system features multiple advantages, such as easy preparation, broad compatibility of functional groups, fast response with high release yield and tunable emission spectra, and is expected to have broad applications. 5. The results from the Chapter 3 demonstrsted that the amides of BH derivatives could selectively label proteins vicinal dithiol. We synthesized a series of BH derivatives scaffold by the Ugi reaction. Then we evaluated the antitumor activity of the BH compounds and found that compound 5f has a good inhibition on the growth of HepG 2 cells. Further mechanistic studies disclose that 5g may selectively inhibit Trx system and ultimately induce inhibition of HepG 2 cells.
Document Type学位论文
First Author AffilicationCollege of Chemistry and Chemical Engineering
Recommended Citation
GB/T 7714
赵澜宁. Baylis–Hillman类荧光探针/药物的设计合成及生物应用[D]. 兰州. 兰州大学,2021.
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