Ojwang J O, Mustain S D, Marshall H B, Rao T S, Chaudhary N, Walker D A, Hogan M E, Akiyama T, Revankar G R, Peyman A, Uhlmann E, Rando R F
Aronex Pharmaceuticals, Inc., The Woodlands, Texas 77381-4223, USA.
Biochemistry. 1997 May 20;36(20):6033-45. doi: 10.1021/bi970124x.
Tumor necrosis factor alpha (TNF alpha), a polypeptide produced by activated macrophages, is a highly pleiotropic cytokine which elicits inflammatory and immunological reactions. The binding of TNF alpha to tumor necrosis factor receptor type I (TNFRI) is considered the initial step responsible for some of the multiple biological functions mediated by TNF alpha. The role of TNF alpha as an inflammatory mediator through human TNFRI makes TNFRI an attractive target for intervention in both acute and chronic inflammatory diseases. In this study, we have identified partial phosphorothioate oligodeoxyribonucleotides (ODNs) containing C-5 propynyl or hexynyl derivatives of 2'-deoxyuridine which specifically inhibited TNFRI and subsequently inhibited the functions of TNF alpha mediated through TNFRI. The most active ODNs were directed against the 3'-poly adenylation signal site on the TNFRI mRNA, and in a cellular assay, gene-specific antisense inhibition occurred in a dose-dependent fashion at submicromolar concentrations, in the presence of Cellfectin. The inhibition of gene expression correlated with the binding affinity of the ODN for the target mRNA. The ODNs lowered TNFRI protein levels and TNF alpha-mediated functions by specifically reducing levels of TNFRI mRNA. These anti-TNFRI ODNs offer a novel approach for controlling biological functions of TNF alpha and may be useful as human therapeutic agents for treating diseases in which TNF alpha has been implicated.
肿瘤坏死因子α（TNFα）是一种由活化巨噬细胞产生的多肽，是一种具有高度多效性的细胞因子，可引发炎症和免疫反应。TNFα与I型肿瘤坏死因子受体（TNFRI）的结合被认为是介导TNFα多种生物学功能的某些作用的起始步骤。TNFα通过人TNFRI作为炎症介质的作用使得TNFRI成为干预急性和慢性炎症性疾病的一个有吸引力的靶点。在本研究中，我们鉴定了含有2'-脱氧尿苷的C-5丙炔基或己炔基衍生物的部分硫代磷酸酯寡脱氧核糖核苷酸（ODN），其特异性抑制TNFRI，随后抑制通过TNFRI介导的TNFα的功能。最具活性的ODN靶向TNFRI mRNA上的3'-聚腺苷酸化信号位点，并且在细胞试验中，在存在Cellfectin的情况下，基因特异性反义抑制以亚微摩尔浓度呈剂量依赖性发生。基因表达的抑制与ODN对靶mRNA的结合亲和力相关。这些ODN通过特异性降低TNFRI mRNA水平降低了TNFRI蛋白水平和TNFα介导的功能。这些抗TNFRI ODN为控制TNFα的生物学功能提供了一种新方法，并且可能作为治疗与TNFα相关疾病的人类治疗剂有用。
