DEVELOPMENT OF A THERAPEUTIC TNF DEGRADATION PRODUCT

治疗性 TNF 降解产品的开发

• 批准号: 3493363
• 负责人: John P Fruehauf
• 金额: $ 5万
• 依托单位: ONCOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-06-01 至 1993-12-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3493363
• 关键词: MCF7 cell; antineoplastics; breast neoplasms; colon neoplasms; cytotoxicity; drug design /synthesis /production; drug resistance; enzyme linked immunosorbent assay; female; high performance liquid chromatography; human tissue; lung neoplasms; melanoma; multiple myeloma; neoplasm /cancer pharmacology; nonHodgkin's lymphoma; ovary neoplasms; peptide chemical synthesis; protein sequence; radiotracer; stainings; tissue /cell culture; tumor necrosis factor alpha

Recombinant human tumor necrosis factor-alpha (TNF) has had only modest success in clinical trials, in part because of dose limiting toxicities. To more fully realize TNF's clinical activity it will be necessary to develop more potent TNF analogues possessing fewer untoward effects. Several studies have found that TNF cytotoxicity depends on TNF internalization and processing by the cell. We compared the processing of TNF by TNF-sensitive and resistant cancer cells of breast and prostate origins. Sensitive cells were found to process internalized TNF to 15 kD and 5.5 kD species, while the resistant cell lines produced multiple lower molecular weight TNF products. We also evaluated the toxicity of conditioned medium from sensitive cells to determine if media containing the degradation products was toxic to resistant cells. Hydrophobic interaction column HPLC was employed to isolate and purify the degradation products for evaluation of their cytotoxic activity. Conditioned media and the HPLC fractions of conditioned media enriched for the 5.5 kD TNF fragment were toxic to both TNF-sensitive and -resistant tumor cells. Phase I of this project would isolate, sequence and synthesize the 5.5 kD degradation product and assay its antineoplastic activity against fresh human tumors of various histology's. Phase II of this project would evaluate the toxicity profile and anti-tumor action of this peptide in vivo in murine models if Phase I results are encouraging. The ultimate goal would be to develop a new chemotherapeutic agent.

重组人肿瘤坏死因子-α（TNF）仅具有适度的 在临床试验中取得成功，部分原因是剂量限制性毒性。 为了更充分地认识TNF的临床活性，有必要 开发具有更少不良反应的更有效的TNF类似物。 几项研究发现，TNF细胞毒性取决于TNF 由细胞内化和加工。 我们比较了 乳腺癌和前列腺癌的肿瘤坏死因子敏感和耐药细胞的肿瘤坏死因子 起源. 发现敏感细胞将内化的TNF加工至15 kD 和5.5 kD的物质，而抗性细胞系产生多个 低分子量TNF产物。 我们还评估了 条件培养基，以确定是否含有 降解产物对抗性细胞有毒。 疏水 采用相互作用柱高效液相色谱法分离和纯化 降解产物，以评价其细胞毒性活性。 条件培养基和富集的条件培养基的HPLC级分 对于5.5kD的TNF片段，对TNF敏感的和 - 抗性肿瘤细胞。 这个项目的第一阶段将分离，测序 并合成5.5kD降解产物， 抗各种新鲜人肿瘤的抗肿瘤活性 组织学。 该项目的第二阶段将评估 在鼠模型中，如果 第一阶段的结果令人鼓舞。 最终目标是开发一个 新的化疗药物。