Synthesis and DNA binding affinity evaluation of novel gilvocarcin-C-glucosides

新型吉沃卡星-C-葡萄糖苷的合成及DNA结合亲和力评价

基本信息

批准号：
8473229
负责人：
Thomas Gerard Minehan
金额：
$ 10.49万
依托单位：
CALIFORNIA STATE UNIVERSITY NORTHRIDGE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-06-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8473229
关键词：
2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole Acids Address Affinity Alcohols Binding Binding Sites Biological Factors Bistris Carbohydrates Carbon Carboxylic Acids Characteristics Code Complex Coupling Cytotoxic agent DNA DNA Binding DNA Binding Agent DNA Minor Groove Binding DNA Sequence DNA Sequence Rearrangement Data Development Disaccharides Disease Esterification Evaluation Excision Family Fluorescence Spectroscopy Future Gene Targeting Genes Genetic Transcription Glucose Glucosides Glycosides Goals Health Intercalating Agents Iodination reaction Ligands Link Literature Measures Mediating Minor Groove Mono-S Monosaccharides Naphthalene Nature Nucleotides Palladium Pharmaceutical Preparations Poly dA-dT Positioning Attribute Preparation Process Protocols documentation Reaction Reagent Rebeccamycin Regulation Relative (related person)Research Series Site Specificity Techniques Therapeutic Agents Titrations Toxic effect Trisaccharides Vertebral column antineoplastic antibiotics calf thymus DNA cancer cell cancer therapy cellular targeting chromophore cytotoxicity design desulfurization flexibility gilvocarcin M glycosylation hedamycin member neoplastic cell novel oxidation preference promoter research study small molecule sugar ultraviolet

项目摘要

DESCRIPTION (provided by applicant): Numerous natural products that are known to bind duplex DNA possess two key structural characteristics that allow tight binding interactions with their cellular target: a planar aromatic aglycone that intercalates the backbone of DNA, and one or more carbohydrate moieties that participate in non-covalent interactions with the major and/or minor grooves. C-Aryl glycosides, substances containing carbon-carbon bonds between an aromatic intercalator and one or more carbohydrate residues, may thus be viewed as promising candidates for the development of high affinity DNA-binding agents. Although it has been shown that aromatic intercalators containing attached glycosyl residues have a superior ability to bind DNA in comparison to intercalators lacking such residues, there exist in the literature no systematic studies quantitatively comparing the DNA binding affinity of intercalators containing one, two, or three appended glycosyl units. In the proposed research, we will prepare a series of mono-, bis-, and tris-C-aryl glucoside derivatives of the gilvocarcin M chromophore and evaluate their association constants for duplex DNA by fluoresence and ultraviolet spectrocopic techniques. Members of the gilvocarcin family of C-aryl glycoside natural products are ideal templates for undertaking such a study since they are known DNA-binding agents that possess high antitumor activity with low overall toxicity. The synthesis plan calls for the convergent assembly of protected carbohydrate, naphthalene, and o-iodobenzoic acid subunits, and will allow the preparation of four gilvocarcin C-glucoside derivatives in seven to nine steps. The key carbon-carbon bond-forming reactions between aromatic and carbohydrate moieties will involve the direct coupling of arylmetal (aryllithium or arylcuprate) reagents and sugar thiolactones, followed by stereoselective radical reduction of the intermediate hemithioketal. The proposed binding studies measuring the strength of interaction of our gilvocarcin derivatives with calf-thymus DNA, poly(dA/dT)7poly(dA/dT), and poly(dG/dC)7poly(dG/dC) will not only allow us to assess the degree to which the attachment of additional carbohydrates to the gilvocarcin chromophore alters DNA binding affinity, but also will allow us to establish a correlation between the site of carbohydrate attachment on the chromophore (corresponding to placement in the major and/or minor grooves) and DNA binding affinity; furthermore, the DNA sequence preferences of our derivatives will also be apparent from the data obtained. Since it has been hypothesized that the DNA sequence specificity of C-aryl glycoside natural products depends on the nature of the glycosidic substituents, these studies will pave the way for the eventual preparation of gilvocarcin derivatives bearing a variety of different carbohydrate moieties useful in establishing paradigms for the design of sequence-dependent DNA-binding ligands. Such molecules, if capable of selectively interacting with the promoters or coding regions of target genes, may ultimately be used for the regulation of gene transcription and as therapeutic agents for a wide range of disease states.

描述（由申请人提供）：已知结合双链 DNA 的许多天然产物具有两个关键的结构特征，可与其细胞靶标紧密结合相互作用：插入 DNA 主链的平面芳香苷元，以及插入 DNA 骨架的一个或多个碳水化合物部分。参与与主要和/或次要凹槽的非共价相互作用。 C-芳基糖苷是在芳香族嵌入剂和一个或多个碳水化合物残基之间含有碳-碳键的物质，因此可以被视为开发高亲和力 DNA 结合剂的有希望的候选物。尽管已经表明，与缺乏此类残基的嵌入剂相比，含有附着的糖基残基的芳香族嵌入剂具有更优异的结合DNA的能力，但文献中还没有系统性的研究来定量比较含有一个、两个或三个的嵌入剂的DNA结合亲和力。附加糖基单元。在本研究中，我们将制备一系列 gilvocacin M 发色团的单、双和三 C 芳基葡萄糖苷衍生物，并通过荧光和紫外光谱技术评估它们与双链 DNA 的缔合常数。 C-芳基糖苷天然产物 gilvocacin 家族的成员是进行此类研究的理想模板，因为它们是已知的 DNA 结合剂，具有高抗肿瘤活性和低总体毒性。该合成计划要求将受保护的碳水化合物、萘和邻碘苯甲酸亚基聚合组装，并将允许通过七到九步制备四种吉沃卡星C-葡萄糖苷衍生物。芳香族和碳水化合物部分之间关键的碳-碳键形成反应将涉及芳基金属（芳基锂或芳基铜酸盐）试剂和糖硫内酯的直接偶联，然后对中间体半硫缩酮进行立体选择性自由基还原。拟议的结合研究测量了我们的 gilvocacin 衍生物与小牛胸腺 DNA、poly(dA/dT)7poly(dA/dT) 和 poly(dG/dC)7poly(dG/dC) 的相互作用强度，这不仅使我们能够评估附加碳水化合物与 gilvocacin 发色团的结合改变 DNA 结合亲和力的程度，而且还将使我们能够建立碳水化合物附着位点之间的相关性发色团（对应于大沟和/或小沟中的位置）和 DNA 结合亲和力；此外，我们的衍生物的 DNA 序列偏好也将从获得的数据中显而易见。由于假设 C-芳基糖苷天然产物的 DNA 序列特异性取决于糖苷取代基的性质，因此这些研究将为最终制备带有各种不同碳水化合物部分的 gilvocacin 衍生物铺平道路，这些衍生物可用于建立范例用于设计序列依赖性 DNA 结合配体。此类分子如果能够选择性地与靶基因的启动子或编码区相互作用，则最终可用于基因转录的调节并作为多种疾病状态的治疗剂。