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

• 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
• 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

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）：已知结合双链体DNA的许多天然产物具有两个关键的结构特征，其允许与其细胞靶标的紧密结合相互作用：嵌入DNA骨架的平面芳香糖苷配基，以及参与与大沟和/或小沟的非共价相互作用的一个或多个碳水化合物部分。C-芳基糖苷，在芳香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，因此可以被视为用于开发高亲和力DNA结合剂的有希望的候选物。尽管已经表明，含有连接的糖基残基的芳族嵌入剂与缺乏这种残基的嵌入剂相比具有上级的结合DNA的能力，但在文献中没有定量比较含有一个、两个或三个连接的糖基单元的嵌入剂的DNA结合亲和力的系统研究。在本研究中，我们将制备一系列gilvocarcin 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-芳基糖苷天然产物的gilvocarcin家族的成员是进行这种研究的理想模板，因为它们是已知的具有高抗肿瘤活性和低总体毒性的DNA结合剂。该合成计划要求保护的碳水化合物，萘，和邻碘苯甲酸亚基的会聚组装，并将允许在七至九个步骤中制备四个gilvocarcin C-葡萄糖苷衍生物。芳香族和碳水化合物部分之间的关键碳-碳键形成反应将涉及芳基金属（芳基锂或芳基铜酸盐）试剂和糖硫代内酯的直接偶联，随后是中间体半硫代缩酮的立体选择性自由基还原。测量我们的gilvocarcin衍生物与小牛胸腺DNA、poly（dA/dT）7 poly（dA/dT）和poly（dG/dC）7 poly（dG/dC）的相互作用强度的所提出的结合研究将不仅允许我们评估额外的碳水化合物与gilvocarcin发色团的连接改变DNA结合亲和力的程度，还能让我们建立发色团上碳水化合物附着位点（对应于在大沟和/或小沟中的放置）和DNA结合亲和力;此外，我们的衍生物的DNA序列偏好也将从所获得的数据中显而易见。由于已经假设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具有多种不同碳水化合物部分的gilvocarcin衍生物铺平道路，这些碳水化合物部分可用于建立序列依赖性DNA结合配体的设计范例。这些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和作为广泛疾病状态的治疗剂。