Development of screening systems for new anti-tumor agents using antibiotic-binding proteins and sensor-promoters

使用抗生素结合蛋白和传感器启动子开发新型抗肿瘤药物的筛选系统

基本信息

批准号：
07556093
负责人：
SUGIYAMA Masanori
金额：
$ 1.22万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

Bleomycin (Bm) has been used in the combination chemotherapy based treatment of carcinoma. But its use is very much limited bccause of its untoward effects of pulmonary toxicity. So, new Bm analogues without htese side effects could be potentially indispensable in the treatment of carcinoma. We constructed a promoter-probe vector, designated pMX180, carrying a Streptomyces tyrosinase genc as a reporter gene. The present study showed that Escherichia coli harboring a plasmid pMX180tac, generated by insertion of tac promoter into pMX180, produced melanin pigment mediated by tyrosinase, when inhibitors against DNA synthesis were added to the culture medium. The Bm family of antibiotics such as pepleomycin, liblomycin and phleomycin were also effective in producing the melanin pigment. Mitomycin C was most effective in the pigment production. Nalidixic acid and azidothymidine also significantly induced the synthesis of melanin pigment. However, some inhibitors of protein and cell wall syntheses did not induce the synthesis of melanin pigment. The culture broth from Bm-producingS.verticillus induced melanin sythesis in this E.coli harboring pMX180tac, suggesting that this plasmied enables visual detection of inhibitors of nucleic acid synthesis that might be used as potent antitumor agents. The vector that we have made enables new Bm analogues to be screened directly in the culture of microorganisms that produce similar anticancer compounds, This system of screeing new Bm analogues, in a very simple, cost effective and time efficient way, would be of great importance in finding a new drug without the hazardous side effects. We call pMX180tac as an intelligent vector. The vector may have application in the efficient screening of microorganism that produce DNA synthesis inhibitors.

博来霉素（BM）已用于基于化学疗法的癌治疗。但是它的使用极为有限，因为它对肺毒性的不良影响。因此，没有HTESE副作用的新的BM类似物对于癌的治疗可能是必不可少的。我们构建了一个指定为PMX180的启动子 - 探针载体，并带有链霉菌酪氨酸酶GenC作为记者基因。本研究表明，当将TAC启动子插入PMX180中产生的具有质粒PMX180TAC的大肠杆菌，当将抑制剂抑制对DNA合成的抑制剂添加到培养基中时，会产生由酪氨酸酶介导的黑色素色素。 BM抗生素的BM家族，例如胸膜霉素，libomycin和phleomycin也有效产生黑色素色素。丝霉素C在色素生产中最有效。奈替二酸和叠氮噻噻替胺还显着诱导了黑色素色素的合成。但是，一些蛋白质和细胞壁合成的抑制剂不会诱导黑色素色素的合成。来自BM产生的培养汤。垂直菌诱导的含有PMX180TAC的大肠杆菌诱导黑色素的结论，这表明这种质量可以视觉检测核酸合成的抑制剂，该抑制剂可能用作有效的抗肿瘤药物。我们已经制造的矢量使新的BM类似物可以直接在产生类似的抗癌化合物的微生物培养中进行筛选，这种散布新的BM类似物的系统以非常简单，具有成本效益的时间和时间高效的方式非常重要，对于在没有危险副作用的情况下寻找一种新药物而言非常重要。我们将PMX180TAC称为智能向量。该载体可能在产生DNA合成抑制剂的微生物的有效筛选中应用。