Cyclic Peptide Protease Inhibitors for the Treatment of Prostate Cancer

用于治疗前列腺癌的环肽蛋白酶抑制剂

• 批准号: 10408279
• 负责人: Aaron Matthew LeBeau
• 金额: $ 18.95万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10408279
• 关键词: Address; Affect; African American; Androgen Receptor; Antiandrogen Therapy; Antibodies; Biochemical; Biodistribution; Biological Assay; Biopsy; Caucasians; Cell Line; Cell surface; Chelating Agents; Chemistry; Clinic; Clinical; Complex; Copper; Cryoelectron Microscopy; Cyclic Peptides; Data; Development; Disease; Dose; Drug Design; Elements; Exhibits; Formalin; Frequencies; Generations; Goals; Human; Immunohistochemistry; In Vitro; Investigation; Label; Lead; Life; Ligand Binding; Malignant neoplasm of prostate; Membrane; Metalloproteases; Metastatic Prostate Cancer; Modality; Molecular Conformation; Monitor; Mus; Nuclear Structure; Organ; Paraffin Embedding; Patients; Peptide Hydrolases; Peptides; Phage Display; Pharmaceutical Preparations; Positron-Emission Tomography; Prostate Adenocarcinoma; Prostate Cancer therapy; Protease Inhibitor; Radiolabeled; Receptor Signaling; Research; Resistance; Screening for Prostate Cancer; Specificity; Structure; Substrate Specificity; Therapeutic; Tissue Embedding; Tissue Microarray; Tissues; Toxic effect; Treatment Efficacy; Treatment Protocols; Tumor Burden; Variant; X-Ray Crystallography; Xenograft Model; Xenograft procedure; alanine aminopeptidase; androgen deprivation therapy; base; bioaccumulation; cancer cell; cancer subtypes; clinical translation; clinically relevant; combat; curative treatments; design; dimer; effective therapy; imaging study; in vitro Model; in vivo; inhibitor/antagonist; innovation; insight; men; molecular imaging; monomer; nanomolar; novel; novel therapeutics; nuclear imaging; overexpression; patient population; peptide drug; prostate cancer cell; prostate cancer model; racial disparity; seryl-histidine; subcutaneous; targeted treatment; therapeutic candidate; therapeutic development; therapeutic target; therapy development; tool; treatment response; tryptophyl-proline; tumor

Although new therapies have had an impact prolonging the quality and quantity of life of men suffering from prostate cancer (PCa), novel treatments are urgently needed to combat aggressive forms of the disease. Recently, we made several key discoveries in the development of therapeutic peptides targeting the membrane bound metalloprotease aminopeptidase N (APN) in aggressive subtypes of PCa. To accurately and reliably interrogate the expression of APN in PCa tissues, we identified a novel antibody by phage display that specifically recognized APN in formalin-fixed paraffin embedded tissues. This antibody allowed us to screen PCa tissue microarrays (TMAs) with a fidelity never before seen. Immunohistochemistry analysis of patient biopsies and TMAs found that APN was preferentially overexpressed in non-androgen receptor (AR) driven PCa compared to AR-driven prostate adenocarcinoma. We also documented in a race disparity TMA and patient biopsies that APN was expressed in the PCa of African-American men while absent in Caucasian patients. Both African- American men and men with non-AR driven PCa represent patient populations who exhibit aggressive disease with poor overall survival. Using a rational design approach based off of the substrate specificity of APN, we identified a cyclic peptide (Leu-His-Ser-Pro-Trp = cLHSPW) that was a micromolar inhibitor of APN’s enzymatic activity. This peptide was found to be moderately therapeutic in APN-expressing PCa xenografts at high doses. Tethering of two cLHSPW peptides together by a linker created a dimer with enhanced potency in vitro and in vivo. The dimer was a nanomolar inhibitor of APN and was rapidly internalized by the cancer cell. When administered at the same high dose as the monomer, the dimer led to complete tumor attrition in vivo. These findings suggest that higher order multivalent cLHSPW peptides may be effective APN-targeted therapeutics. In this proposal, we will synthesize dimeric, trimeric, and tetrameric cLHSPW peptides using macrocyclic frames. In addition to ease of synthesis, the macrocyclic frames can also chelate radiometals for positron-emission tomography (PET). To develop a new class of PCa therapeutics, the in vitro therapeutic efficacy of the peptides will be characterized and PET imaging will be performed in vivo to monitor biodistribution (Aim 1), the ability of the peptides to eliminate tumor burden in subcutaneous xenografts will be examined (Aim 2) and X-ray crystallography and cryo-electron microscopy will be performed on APN complexed with the peptides to obtain mechanistic insights and ultimately lead to the development of optimized inhibitors (Aim 3). Our preliminary data strongly suggest that we have developed potent therapeutics that have the potential to result in a dramatic shift in how aggressive PCa is treated.

尽管新疗法产生了影响，从而延长了患有男性的生活的质量和数量 前列腺癌（PCA），迫切需要新的治疗方法来打击侵略性的疾病。 最近，我们在靶向膜的热肽的发展方面做出了一些关键发现 PCA侵袭性亚型中结合的金属蛋白酶N（APN）。准确可靠地 询问APN在PCA组织中的表达，我们通过噬菌体显示了一种新颖的抗体 在福尔马林固定的石蜡嵌入组织中识别的APN。这种抗体使我们能够筛选PCA组织 以前从未见过的微阵列（TMA）。患者活检和 TMAS发现，与非雄激素受体（AR）驱动的PCA相比，APN优先表达 AR驱动的前列腺腺癌。我们还记录了种族差异TMA和患者活检 APN在非裔美国人的PCA中表达，而白人患者缺席。两个非洲 - 非AR驱动PCA的美国男性和男性代表暴露侵略性疾病的患者人群 总体生存率差。使用基于APN的底物特异性的合理设计方法，我们 鉴定出是APN酶促的微摩尔抑制剂的环状肽（Leu-His-Ser-ser-pro-trp = clhspw） 活动。发现这种胡椒在高剂量的表达APN的PCA异种移植物中具有适度的治疗性。 将两个CLHSPW Pepperides链接在一起，由一个接头创建了一个二聚体，并具有增强的体外效力 体内。该二聚体是APN的纳摩尔抑制剂，被癌细胞迅速内化。什么时候 二聚体以与月份相同的高剂量给药，导致体内完全肿瘤损耗。这些 研究结果表明，高阶多价CLHSPW Petides可能是有效的APN靶向疗法。在 该建议，我们将使用大环框架合成二聚体，三聚体和四聚体CLHSPW肽。 除了易于合成之外，大环框架还可以珍惜辐射标准的辐射。 断层扫描（宠物）。为了开发新的PCA疗法，Petides的体外疗法效率 将在体内进行表征，并将在体内进行宠物成像（AIM 1）， 将检查消除皮下异种移植物中肿瘤燃烧的肽（AIM 2）和X射线 晶体学和冷冻电子显微镜将在与Petides复合的APN上进行 机械洞察力，最终导致了优化抑制剂的发展（AIM 3）。我们的初步数据 强烈建议我们开发了潜在的疗法，有可能导致急剧转变 在如何治疗侵略性的PCA中。