Identification of pancreatic cancer specific high affinity targeting ligands

胰腺癌特异性高亲和力靶向配体的鉴定

• 批准号: 7297106
• 负责人: Pappanaicken Rangasamy Kumar
• 金额: $ 7.6万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7297106
• 关键词: Adverse effects; Affinity; Amino Acids; Antibodies; Apoptosis; Ascites; Binding; Biological; Blood; Cancer Patient; Cancer cell line; Cell surface; Clinic; Clinical Trials; Collaborations; Collection; Coupling; Cyclic Peptides; Data; Defect; Drug Delivery Systems; Drug Industry; Drug resistance; Ductal Epithelial Cell; Early Diagnosis; Evaluation; Feces; Flow Cytometry; Goals; Grant; Hematologist; Hepatocyte; Human; Image; Lead; Letters; Libraries; Life; Ligand Binding; Ligands; Link; Liquid substance; Magic; Malignant Neoplasms; Malignant neoplasm of pancreas; Medical; Medical center; Methods; Monoclonal Antibodies; Mus; Normal Cell; Numbers; Oncologist; Outcome Study; Pancreas; Peptides; Pharmaceutical Preparations; Pleural; Radiation therapy; Radio; Range; Reticuloendothelial System; Scientist; Screening procedure; Signal Pathway; Solid Neoplasm; Source; Specificity; Standards of Weights and Measures; Structure; Structure-Activity Relationship; Therapeutic; Therapeutic Agents; Time; Tissue Microarray; Toxin; Universities; Writing; Xenograft Model; base; cancer cell; chemotherapy; circulating cancer cell; combinatorial; design; gemcitabine; high throughput screening; killings; mouse model; novel; novel therapeutics; peptidomimetics; receptor; research study; size; small molecule; tumor

DESCRIPTION (provided by applicant): Pancreatic cancer (PCa) is one of the most lethal malignancies in humans. Gemcitabine is the current standard chemotherapy of advanced PCa but it is still far from optimam and novel therapeutic strategies are urgently needed. Accumulating clinical-trial results are showing that most tumors, particularly solid tumors are multifactorial and are frequently linked to defects in more than one signaling pathway. Therefore, a dual targeting or multi-targeting therapy might be more rational, not only to efficiently eliminate cancer cells, but also to limit the emergence of drug resistance. A method of identifying novel targeting ligands is much warranted. High throughput screening (HTS) of compound collections, derived from natural as well as synthetic sources have been used by the pharmaceutical industry for the identification of drug leads. To make HTS more efficient we used "one-bead one-compound" (OBOC) combinatorial library method by which up to 75,000 compounds can be screened in 10 cm petri dish in parallel. OBOC library is generated in a unique way of having only one type of compounds on each bead of ~ 100 micron size. It is derived by random split-mix synthesis by coupling various building blocks such as L-amino acids, D-amino acids, unnatural amino acids and small molecules or mixture of all. The generated OBOC libraries can be screened against a variety of biological targets, including live cancer cells. In the past year, we have used three pancreatic cancer cell lines to screen several OBOC combinatorial libraries and identified several D-amino acid containing cyclic peptides that bind preferentially to pancreatic cancer cells but not to immortalized normal hepatocytes. These targeting ligands showed specific binding in ¿M range and not inducing apoptosis in cancer cells. However these targeting ligands can be an excellent lead compounds to derive more structure assisted combinatorial libraries to identify high affinity ligands that bind pancreatic cancer cells specifically. These high affinity ligands can be used for imaging and to trap circulating cancer cells in the biological fluids such as blood, pancreatic ascites and stool. In this proposal, we plan to exploit these ligands by developing more focused OBOC libraries, in which low passage pancreatic cancer cell lines and normal immortalized pancreatic ductal cells will be used to screen for high affinity ligands in an ultra-high-throughput manner. The over all aim of this proposal is to identify novel pancreatic cancer-specific targeting ligands that can be used as effective imaging and as drugs delivery agents to treat pancreatic cancer. The specific aims of this proposal are as follows: Aim 1. To design and synthesize six focused OBOC libraries, from the lead compound cKMIMRA and cVVKMHKc Aim 2.To identify high affinity pancreatic cancer specific ligands by high-throughput screening OBOC libraries from aim 1. Aim 3. To evaluate specificity and high affinity binding of selected ligands by tissue microarray and flowcytometry analysis. Time line 12 months 24 months Aim1 To synthesize several OBTC libraries Aim2 HTS stringent screening of high affinity ligands Aim3 Evaluation of specificity and high affinity of selected ligands The out come of this study will generate pancreatic cancer specific high affinity ligands that will be used to write a R21-grants for imaging pancreatic cancer for early detection in collaboration Dr. Maitra, john Hopkins University (letter attached) and identification circulating cancer cells in pancreatic cancer patients in collaboration with Dr. Shiro Uriyama, gasteroenterologist at UC Davis Medical Center at Sacramento. There will be enough data generated through this grant to write an R01-grant to evaluate therapeutic potential of these high-affinity ligands in combination with or without radiometal/ chemotoxin in mouse model and characterization of targeting receptors in pancreatic cancer cells. Currently available conventional chemotherapy and radiotherapy will not differentiate in a great way between normal cells and cancer cells thereby lot of side effects to the body. Scientists are looking various ways to kill cancer cells without harming normal cells. In this research study our goal is to identify targeting ligands (Magic bullets) that are specifically target pancreatic cancer cells and not the normal cells. This is achieved by screening few thousand to million compounds at the same time against the cancer cells by recently developed one-bead-one-compound technolology. These targeting ligands can be used to deliver drugs, toxins or radiometal specifically to cancer cells and not the normal cells.

描述（由适用提供）：胰腺癌（PCA）是人类最致命的恶性肿瘤之一。吉西他滨是晚期PCA的当前标准化学疗法，但远离最佳状态，迫切需要新颖的治疗策略。累积的临床试验结果表明，大多数肿瘤，尤其是实体瘤是多因素的，并且经常与多个信号通路中的缺陷有关。因此，双重靶向或多目标疗法可能更合理，不仅可以有效消除癌细胞，而且还可以限制耐药性的出现。识别新颖靶向配体的一种方法是非常保证的。制药行业已经使用了自然和合成来源的化合物集合的高吞吐量筛选（HTS），用于鉴定药物铅。为了使HTS更加高效，我们使用了“单孔单杂色”（OBOC）组合库方法，通过该方法可以在10 cm Petri菜肴中筛选高达75,000种化合物。 OBOC库是以独特的方式生成的，即在〜100微米尺寸的每个珠子上只有一种类型的化合物。它是通过将各种构件（例如L-氨基酸，D-氨基酸，非天然氨基酸，小分子或所有混合物）偶联而得出的。可以针对包括活癌细胞在内的各种生物学靶标筛选生成的OBOC文库。在过去的一年中，我们使用了三种胰腺癌细胞系来筛选几个OBOC组合文库，并确定了几种含有含有环状肽的D-氨基酸，这些肽优先与胰腺癌细胞结合，但不能与永生化的正常肝细胞结合。这些靶向配体在``M范围''中显示出特异性的结合，而在癌细胞中未诱导凋亡。但是，这些靶向配体可能是一种出色的铅化合物，可以得出更多结构辅助组合文库，以鉴定特定结合胰腺癌细胞的高亲和力配体。这些高亲和力配体可用于成像和捕获诸如血液，胰腺腹水和粪便等生物学液中的循环癌细胞。在此提案中，我们计划通过开发更集中的OBOC库来探索这些配体，其中低通道胰腺癌细胞系和正常永生的胰腺导管细胞将用于以超高直射的方式筛选高亲和力的高度配体。该提案的所有目的是确定可以用作有效成像的新型胰腺癌特异性靶向配体，并用作治疗胰腺癌的药物递送剂。该提案的具体目的如下：目的1。从铅化合物CKMIMRA和CVVKMHKC AIM 2.识别高亲和力胰腺癌特异性配体，通过从AIM AIM 1进行评估的lig lig iganders 3。流程仪分析。 Time line 12 months 24 months Aim1 To synthesize several OBTC libraries Aim2 HTS stringent screening of high affinity ligands Aim3 Evaluation of specificity and high affinity of selected ligands The out come of this study will generate pancreatic cancer specific high affinity ligands that will be used to write a R21-grants for imaging pancreatic cancer for early detection in collaboration Dr. Maitra, john Hopkins University (letter附有）和鉴定胰腺癌患者的循环癌细胞与萨克拉曼多加州大学戴维斯分校医学中心的Gasteroenteroloks Shiro Uriyama博士合作。将有足够的数据通过该赠款生成，以编写R01赠品，以评估这些高亲和力配体的治疗潜力，并在小鼠模型中与有或没有放射性/趋化毒素结合使用，并在胰腺癌细胞中表征靶向受体。当前可用的常规化疗和放射疗法不会以正常细胞和癌细胞之间的良好方式分化，从而对人体有很大的副作用。科学家正在寻找多种杀死癌细胞而不会损害正常细胞的方法。在这项研究中，我们的目标是鉴定针对靶向胰腺癌细胞而不是正常细胞的靶向配体（魔术斗牛）。这是通过在最近开发的一珠一对组合技术对癌细胞同时筛选几千万用化合物来实现的。这些靶向配体可用于将药物，毒素或辐射量专门用于癌细胞，而不是正常细胞。