Preclinical development of AlaTIMP-2 as an cancer therapeutic

AlaTIMP-2 作为癌症治疗剂的临床前开发

• 批准号: 8157696
• 负责人: William Stetler-Stevenson
• 金额: $ 94.14万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8157696
• 关键词: Development; Malignant Neoplasms; Therapeutic; pre-clinical

Working towards our stated goal of preclinical development of TIMP-2 as a novel cancer therapeutic we have performed in vivo xenograft growth experiments. These experiments have shown that the TIMP-2 homolog lacking MMP inhibitory activity known as Ala+TIMP-2 effectively inhibits growth of the human A549 lung cancer xenograft, when overexperssed in the tumor cells using a retroviral vector system. Ongoing experiments are utilizing the TET-On system to examine the effect of forced expression of TIMP-2 and Ala+TIMP-2 in established tumor systems. To further the development of Ala+TIMP-2 as a therapeutic we have developed CHO cell expression systems for the production of recombinant Ala+TIMP-2 and TIMP-2, and are continuing these efforts to optimize production of large quantities under good manufacturing procedures (GMP). It is our plan to test these materials as therapeutic agents in both xenogfraft and syngeneic murine tumor models to demonstrate the effectiveness of treatment with exogenous recombinant proteins. We also plan to utilize newly developed genetically engineered mouse models (GEMMs) of lung cancer for testing the anti-tumor effects of recombinant exogenous TIMP-2 and Ala+TIMP-2. The results will be compared with the effects of other members of the TIMP family for potency and efficacy. These models will focus on the treatment of lung cancer (A549 and Lewis Lung) as well as melanoma (A2058 and B16F10). Various dosing regimens will be utilized to compare the relative in vivo effectiveness of Ala+TIMP-2 compared to TIMP-2. Preliminary studies indicate that Ala+TIMP-2 is more effective than TIMP-2, which is attributed to the fact that Ala+TIMP-2 does not bind to the active site of MMP like TIMP-2, therefore effectively increasing its concentration for cell binding sites. Another important aspect of this project is to determine if we can develop peptide analogs that could be utilized for in vivo therapy. Furthermore we propose to develop a high throughput screening assay to screen synthetic small molecule analogs that can compete for TIMP-2, Ala+TIMP-2, or TIMP peptide binding to the cell surface receptor integrin alpha3 beta1, that we have shown modulates the anti-angiogenic and anti-tumorigenic activity of Ala+TIMP-2.

为了实现我们所述的TIMP-2作为新型癌症治疗剂的临床前开发目标，我们进行了体内异种移植物生长实验。这些实验已经表明，当使用逆转录病毒载体系统在肿瘤细胞中过表达时，称为Ala+TIMP-2的缺乏MMP抑制活性的TIMP-2同系物有效地抑制人A549肺癌异种移植物的生长。正在进行的实验利用TET-On系统来检查TIMP-2和Ala+TIMP-2在已建立的肿瘤系统中的强制表达的效果。为了进一步开发Ala+TIMP-2作为治疗剂，我们已经开发了用于生产重组Ala+TIMP-2和TIMP-2的CHO细胞表达系统，并且正在继续这些努力以优化在良好生产程序（GMP）下的大量生产。我们计划在异种移植和同系小鼠肿瘤模型中测试这些材料作为治疗剂，以证明用外源重组蛋白治疗的有效性。我们还计划利用新开发的肺癌基因工程小鼠模型（GEMMs）来测试重组外源性TIMP-2和Ala+TIMP-2的抗肿瘤作用。将结果与TIMP家族其他成员的效力和疗效进行比较。这些模型将专注于肺癌（A549和刘易斯肺）以及黑色素瘤（A2058和B16 F10）的治疗。将使用各种给药方案来比较Ala+TIMP-2与TIMP-2相比的相对体内有效性。初步研究表明，Ala+TIMP-2比TIMP-2更有效，这归因于Ala+TIMP-2不像TIMP-2那样结合MMP的活性位点，因此有效地增加了其用于细胞结合位点的浓度。该项目的另一个重要方面是确定我们是否可以开发可用于体内治疗的肽类似物。此外，我们建议开发高通量筛选测定以筛选合成的小分子类似物，其可以竞争TIMP-2、Ala+TIMP-2或TIMP肽与细胞表面受体整合素α 3 β 1的结合，我们已经显示整合素α 3 β 1调节Ala+TIMP-2的抗血管生成和抗肿瘤发生活性。