Multimeric Ligands for Targeting Melanoma

用于靶向黑色素瘤的多聚配体

• 批准号: 8288314
• 负责人: Victor J Hruby
• 金额: $ 60.07万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-15 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8288314
• 关键词: ADRB1 gene; Adrenergic Receptor; Affinity; Agonist; Antigens; Avidity; Behavior; Binding; Biodistribution; Biological Assay; Biological Models; Cell Surface Proteins; Cell Surface Receptors; Cell surface; Cells; Characteristics; Chemistry; Cholecystokinin; Complex; Confusion; DNA Microarray Chip; Data; Detection; Disease; Drug Kinetics; Engineering; Epitopes; Equilibrium; Fluorescence; G-Protein-Coupled Receptors; Generations; Goals; HCT116 Cells; Image; Immunohistochemistry; In Vitro; Individual; Label; Length; Lesion; Libraries; Ligands; Link; Literature; Malignant - descriptor; Measures; Melanocortin 1 Receptor; Metastatic Lesion; Metastatic Melanoma; Modeling; Molecular; Neoplasm Metastasis; Normal tissue morphology; Patients; Pattern; Research; Screening procedure; Solutions; Specificity; Stream; System; Testing; Time; Tissue Microarray; Tissues; Validation; Work; analytical method; base; cDNA Arrays; cancer cell; crosslink; cyanine dye 5; design; fMet-Leu-Phe receptor; flexibility; fluorophore; improved; in vivo; melanocortin receptor; melanoma; molecular phenotype; monomer; overexpression; programs; prospective; receptor; receptor density; research study; single photon emission computed tomography; stoichiometry; targeted delivery; tumor; ADRB1 gene; Adrenergic Receptor; Affinity; Agonist; Antigens; Avidity; Behavior; Binding; Biodistribution; Biological Assay; Biological Models; Cell Surface Proteins; Cell Surface Receptors; Cell surface; Cells; Characteristics; Chemistry; Cholecystokinin; Complex; Confusion; DNA Microarray Chip; Data; Detection; Disease; Drug Kinetics; Engineering; Epitopes; Equilibrium; Fluorescence; G-Protein-Coupled Receptors; Generations; Goals; HCT116 Cells; Image; Immunohistochemistry; In Vitro; Individual; Label; Length; Lesion; Libraries; Ligands; Link; Literature; Malignant - descriptor; Measures; Melanocortin 1 Receptor; Metastatic Lesion; Metastatic Melanoma; Modeling; Molecular; Neoplasm Metastasis; Normal tissue morphology; Patients; Pattern; Research; Screening procedure; Solutions; Specificity; Stream; System; Testing; Time; Tissue Microarray; Tissues; Validation; Work; analytical method; base; cDNA Arrays; cancer cell; crosslink; cyanine dye 5; design; fMet-Leu-Phe receptor; flexibility; fluorophore; improved; in vivo; melanocortin receptor; melanoma; molecular phenotype; monomer; overexpression; programs; prospective; receptor; receptor density; research study; single photon emission computed tomography; stoichiometry; targeted delivery; tumor

DESCRIPTION (provided by applicant): The goal of the proposed work is to develop heterobivalent agents for targeted delivery of imaging and therapy to metastatic melanoma. These agents contain two ligands that direct the construct to crosslink two different cell surface receptors, resulting in dramatic increases in binding selectivity. This has advantages over agents directed against single receptors in that they do not rely on overexpression of a single cell surface protein. The proposed constructs also have advantages over other multifunctional agents in that they are produced via convergent synthesis and are designed to be relatively small with favorable ADME characteristics. Work during the first period of support has (1) identified and validated a receptor combination that can be used to target a subset of metastatic melanomas, and (2) demonstrated a proof-of-principle that synthetic heterobivalent agents can crosslink heterologous receptors. These constructs bound with up to 50-fold higher affinity compared to corresponding monovalent interactions. Current research will combine these two advances to develop heterobivalent agents against an identified target receptor pair. To achieve this goal, Aim 1 will use a previously developed G-protein coupled receptor (GPCR) system to derive analytical solutions that predict behavior of multivalent ligands as imaging agents. Aim 2 will develop bivalent agents that target a receptor combination identified in the first period of support: N-formyl peptide receptor like, type 2 (FPRL2) and the type 1 melanocortin receptor (MC1R). In this aim, optimum linker and ligand chemistries will be determined with high-throughput binding assays. Bivalent ligands will be labeled with fluorophores for testing by in-cyto and in-vivo imaging. Those with optimum characteristics will be labeled with DOTA. DOTA derivatives will be used to chelate Eu for ex vivo fluorescence and 111In for in vivo SPECT assessment of pharmacokinetics and biodistribution. Aim 3 will continue the validation effort for 21 additional receptors that were identified as targets during the first period of support. These will be validated primarily though immunohistochemistry of tissue microarrays containing multiple melanoma grades as well as multiple normal tissues. It is expected that this work will result in the validation of additional receptor combinations that will target a majority of metastatic melanomas. At the end of this next period of support, we will have (1) developed more precise analytical methods to predict the behavior of multivalent ligands as imaging agents; (2) developed targeting ligands that will be useful against 40-50% of metastatic melanomas and (3) identified additional 3- and 4-receptor target combinations for the remaining molecular phenotypes of this disease.The goal of the proposed work is to develop platform targeting agents for delivery of imaging and therapy to metastatic melanoma. These agents are heterobivalent in that they link together two ligands that are directed against two different cell surface receptors. Such agents have advantages over agents directed against single epitopes by not relying on a single overexpressed cell surface protein. They also have advantages over other multifunctional agents in that they are produced via convergent synthesis and hence are relatively small with acceptable ADME characteristics.

描述（由申请人提供）：拟议工作的目的是开发用于靶向递送成像和治疗转移性黑色素瘤的异体剂。这些试剂包含两个指导构建体的配体，将构建体与两个不同的细胞表面受体交联，从而导致结合选择性的急剧增加。这比针对单个受体的药物具有优势，因为它们不依赖于单个细胞表面蛋白的过表达。所提出的构建体比其他多功能剂具有优势，因为它们是通过收敛合成生产的，其设计为相对较小，具有有利的ADME特性。在第一阶段的支持中，（1）确定并验证了可用于靶向转移性黑色素瘤的子集的受体组合，（2）证明了合成异体剂可以交叉链接异源受体的原理证明。与相应的单价相互作用相比，这些结构与高达50倍的亲和力结合。当前的研究将结合这两个进步，以开发针对已确定的靶受体对的异体剂。为了实现这一目标，AIM 1将使用先前开发的G蛋白偶联受体（GPCR）系统来得出分析溶液，以预测多价配体作为成像剂的行为。 AIM 2将开发靶向第一个支持中鉴定的受体组合的二价药物：N-甲基肽受体类似，类型2（FPRL2）和1型黑色素质素受体（MC1R）。在此目标中，将使用高通量结合测定法确定最佳连接器和配体化学。二价配体将用荧光团标记，用于通过cyto和体内成像进行测试。那些具有最佳特征的人将用dota标记。 DOTA衍生物将用于螯合欧盟，以进行体内荧光，为111in进行药代动力学和生物分布的体内频谱评估。 AIM 3将继续为21个在支持第一阶段被确定为目标的其他受体的验证工作。这些将主要验证，主要是含有多个黑色素瘤等级以及多个正常组织的组织微阵列的免疫组织化学。预计这项工作将导致验证其他受体组合，该组合将针对大多数转移性黑色素瘤。在下一个支持阶段的结束时，我们将（1）开发更精确的分析方法，以预测多价配体作为成像剂的行为； （2）开发的靶向配体将对40-50％的转移性黑色素瘤有用，（3）确定了该疾病剩余的分子表型的其他3和4受体靶标组合。拟议工作的目标是开发用于递送成像和治疗转移性梅兰瘤的平台靶向药物。这些药物是异常的，因为它们将两个针对两个不同细胞表面受体的配体连接在一起。这种药物与针对单个表位的药物具有不依赖单个过表达的细胞表面蛋白的优势。它们比其他多功能剂具有优势，因为它们是通过收敛合成产生的，因此相对较小，具有可接受的ADME特征。