Search for Reversing Agents of Multidrug-Resistance in Tumor Cells from Marine Organisms

从海洋生物中寻找肿瘤细胞多药耐药性逆转剂

基本信息

批准号：
10680567
负责人：
KOBAYASHI Motomasa
金额：
$ 2.3万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680567/
关键词：
multidrug resistant tumor cells P-glycoprotein multidrug resistance associated protein araguspongines cembrane diterpenes agosterol A KB-C2 cells KB-CV60 cells がん多剤耐性克服物質 multidrug resistance protein Spongia属海綿 glutathione araguspo

项目摘要

In the course of our study of bioactive substances from marine organisms, we focused on a search for reversing agents of multidrug resistance (MDR) in tumor cells. We used two kinds of MDR human epidermoid carcinoma cell lines, KB-C2 cells overexpressing P-glycoprotein (P-gp) and KB-CV60 cells overexpressing multidrug resistance associated protein (MRP) for the screening assay. We found that araguspongines conquered MDR in KB-C2 cells. Araguspongines were isolated from an Okinawan marine sponge of Xestospongia sp. and characterized as macrocyclic dimer of 2,9-disubstituted 1-oxaquinolizidine. Araguspongine D, which is a major component among araguspongines, completely conquered MDR in KB-C2 cells at 10 μg/ml. Araguspongines B and E, which were stereoisomers of araguspongine D, also completely reversed MDR in KB-C2 cells. 5,5'-N-Dimethyl derivative and 2,2'-diol synthesized from araguspongine E did not show reversing activity. We isolated cembrane-type diterpenes reversing MDR in KB-C2 cells from a soft coral of Sinularia sp. This cembranoid completely conquered MDR in KB-C2 cells at 10 μg/ml. This cembranoid is seemed to be an artifact compound, which was produced from known natural cembranolide by addition of n-BuOH. We synthesized a series of cembrane derivatives replaced the n-butyl ester with various alcohols and found that the reversing activity of those derivatives were related to the carbon number of alcohol. Agosterol A, a novel polyhydroxylated sterol acetate, was isolated from a marine sponge of Spongia sp. as a reversing agent of MDR. Agosterol A perfectly reversed resistance to colchitine in KB-C2 cells and resistance to vincristine in KB-CV60 cells at 3μM concentration. Agosterol A recovered the accumulation of vincristine in KB-C2 and KB-CV60 cells to the level equal to that of parental KB 3-1 cells at 3 μM concentration.

在研究海洋生物的生物活性物质的过程中，我们重点寻找肿瘤细胞中多药耐药性（MDR）的逆转剂，我们使用了两种MDR人表皮样癌细胞系，即过度表达P-的KB-C2细胞。糖蛋白 (P-gp) 和过表达多药耐药相关蛋白 (MRP) 的 KB-CV60 细胞进行筛选试验，我们发现阿拉格斯海绵克服了 KB-C2 中的多药耐药性。 Araguspongines 是从 Xestospongia sp. 的冲绳海洋海绵中分离出来的，其特征是 2,9-二取代 1-oxaquinolizidine 的大环二聚体，它是 araguspongines 的主要成分，在 10 时完全克服了 KB-C2 细胞中的 MDR。 μg/ml。阿拉格斯海绵 B 和 E，是阿拉格斯海绵的立体异构体。 D，也完全逆转了KB-C2细胞中的MDR，而从araguspongine合成的5,5'-N-二甲基衍生物和2,2'-二醇在KB-C2中没有显示出逆转MDR的活性。来自 Sinularia sp. 的软珊瑚细胞这种 cembranoid 在 10 μg/ml 下完全克服了 KB-C2 细胞中的 MDR。西松烷内酯似乎是一种人工合成的化合物，它是由已知的天然西松内酯通过添加正丁醇而产生的，我们合成了一系列用各种醇代替正丁酯的西松烷衍生物，并发现这些衍生物的逆转活性与以下因素有关。 Agosterol A 是一种新型多羟基甾醇乙酸酯，是从海绵中分离出来的，作为 MDR 的逆转剂。 3μM浓度的Agosterol A完全逆转了KB-C2细胞对秋水仙碱的耐药性和KB-CV60细胞对长春新碱的耐药性，使KB-C2和KB-CV60细胞中长春新碱的积累恢复到与亲代KB相同的水平。 3-1 个细胞，浓度为 3 μM。