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

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）：拟议工作的目标是开发异二价药物，用于对转移性黑色素瘤进行靶向成像和治疗。这些试剂含有两种配体，可指导构建体交联两种不同的细胞表面受体，从而显着提高结合选择性。这比针对单一受体的药物具有优势，因为它们不依赖于单一细胞表面蛋白的过度表达。所提出的构建体还比其他多功能制剂具有优势，因为它们是通过聚合合成生产的，并且设计得相对较小，具有良好的 ADME 特性。第一个支持期间的工作（1）确定并验证了可用于靶向转移性黑色素瘤子集的受体组合，以及（2）证明了合成异二价药物可以交联异源受体的原理证明。与相应的单价相互作用相比，这些构建体的结合亲和力高出 50 倍。目前的研究将结合这两项进展来开发针对已确定的目标受体对的异二价药物。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) 系统来得出分析解决方案，预测多价配体作为显像剂的行为。目标 2 将开发针对在第一阶段支持期间确定的受体组合的二价药物：N-甲酰肽受体样 2 型 (FPRL2) 和 1 型黑皮质素受体 (MC1R)。为此，将通过高通量结合测定来确定最佳接头和配体化学性质。二价配体将用荧光团标记，以便通过细胞内和体内成像进行测试。那些具有最佳特性的将被贴上DOTA标签。 DOTA 衍生物将用于螯合 Eu，用于离体荧光，并用于螯合 111In，用于体内药代动力学和生物分布的 SPECT 评估。目标 3 将继续验证第一阶段支持期间确定为目标的另外 21 个受体。这些将主要通过包含多种黑色素瘤级别以及多种正常组织的组织微阵列的免疫组织化学来验证。预计这项工作将验证针对大多数转移性黑色素瘤的其他受体组合。在下一阶段的支持结束时，我们将 (1) 开发出更精确的分析方法来预测多价配体作为显像剂的行为； (2) 开发了可用于对抗 40-50% 转移性黑色素瘤的靶向配体，并且 (3) 为该疾病的其余分子表型确定了额外的 3 和 4 受体靶组合。拟议工作的目标是开发用于转移性黑色素瘤成像和治疗的靶向药物平台。这些试剂是异二价的，因为它们将针对两种不同细胞表面受体的两个配体连接在一起。由于不依赖于单一过表达的细胞表面蛋白，此类试剂比针对单一表位的试剂具有优势。与其他多功能试剂相比，它们还具有优势，因为它们是通过聚合合成生产的，因此相对较小，具有可接受的 ADME 特性。