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: 8078218
• 负责人: Thomas Gerard Minehan
• 金额: $ 10.88万
• 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/8078218
• 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; 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. PUBLIC HEALTH RELEVANCE: The development of high-affinity DNA-binding molecules is a crucially important goal in the context of current approaches to cancer therapy; indeed, to maximize tumor cell-specific cytotoxicity, drug molecules must be able to bind tightly to unique targets identified within cancer cells. The proposed research will involve the synthesis and DNA binding-affinity evaluation of a series of mono-, bis-, and tris- C-aryl glucoside derivatives of the gilvocarcins, a class of non-toxic antitumor antibiotics viewed as templates for the design of gene-specific cytotoxic agents. Through this study we hope to establish a correlation between the strength of small molecule-DNA interactions and both the number and position of attachment of carbohydrate residues around an aromatic intercalator.

描述（由申请人提供）：已知会结合双链DNA的许多天然产物具有两个关键的结构特性，可以与其细胞靶标紧密结合相互作用：一种平面芳族芳族性氨基糖酮，可将DNA的骨架相互作用，一个或多个碳水化合物的部分与大型和/或/或或/或或或/或或或或或或/或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或或一位。 c-芳基糖苷，芳香族介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，可以看作是有望发展高亲和力DNA结合剂的有希望的候选者。尽管已经表明，与缺乏此类残基相比，含有附着的糖基残基的芳族介导剂具有较高的结合DNA的能力，但在文献中，没有系统的研究数量地比较了含有一个，二，两个或三个appended糖基单元的Intercalators的DNA结合亲和力。在拟议的研究中，我们将准备一系列的吉尔沃卡菌素和彩色团的单，二芳基糖苷葡萄糖苷衍生物，并通过荧光和紫外线光谱学技术评估其对双链DNA的关联常数。 C-芳基糖苷天然产物的吉尔沃卡素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，具有较高的抗肿瘤活性，总体毒性较低。合成计划要求将受保护的碳水化合物，萘和O-碘苯甲酸亚基的收敛组装，并将允许在七个至九步中制备四种吉尔沃卡菌素C-葡萄糖苷衍生物。芳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（芳基锂或芳基磷酸盐）试剂的直接耦合和糖硫代酮，然后立体选择性地从中间半硫代盐的立体式自由基减少。提出的结合研究测量了我们吉尔沃卡蛋白衍生物与小腿 - 胸腺DNA，poly（da/dt）7poly（da/dt）和聚（DG/DC）7-poly（dg/dc）相互作用的强度我们在发色团（对应于主要和/或次要的凹槽中的放置）和DNA结合亲和力之间建立碳水化合物附着位点之间的相关性；此外，从获得的数据也可以明显看出我们的衍生物的DNA序列偏好。 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.如果这种分子能够选择性地与靶基因的启动子或编码区进行选择性相互作用，则最终可用于调节基因转录，作为广泛疾病状态的治疗剂。 公共卫生相关性：在当前的癌症治疗方法的背景下，高亲和力DNA结合分子的发展是至关重要的目标。实际上，为了最大化肿瘤细胞特异性细胞毒性，药物分子必须能够与癌细胞中鉴定的独特靶标紧密结合。拟议的研究将涉及吉尔沃卡蛋白的一系列单，BIS-和三芳基糖苷衍生物的合成和DNA结合 - 亲和力评估，Gilvocarcins是一类无毒抗肿瘤抗生素，被视为用于基因基因特异性细胞毒素的设计模板的模板。通过这项研究，我们希望建立小分子-DNA相互作用的强度与碳水化合物残基周围的芳香族介入物的数量和位置之间的相关性。