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

• 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

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.

在海洋生物活性物质的研究过程中，我们的重点是寻找肿瘤细胞中多药耐药（MDR）的逆转剂。我们使用过表达p -糖蛋白（P-gp）的KB-C2细胞和过表达多药耐药相关蛋白（MRP）的KB-CV60细胞两种MDR人表皮样癌细胞进行筛选。我们发现，在KB-C2细胞中，蜘蛛碱能够攻克耐多药耐药。从日本冲绳海绵中分离得到一种巨环2,9-二取代- 1-草喹喹啉二聚体。在10 μg/ml的浓度下，阿拉木皂素D完全攻克了KB-C2细胞的MDR。槟榔素B和E是槟榔素D的立体异构体，也能完全逆转KB-C2细胞的MDR。由木棉素E合成的5,5′- n -二甲基衍生物和2,2′-二醇不具有逆转活性。我们从Sinularia sp.软珊瑚中分离出逆转KB-C2细胞MDR的膜型二萜，这种膜类物质在10 μg/ml浓度下完全克服了KB-C2细胞的MDR。这种类胶质物质似乎是一种人造化合物，它是由已知的天然胶质内酯通过添加n-BuOH而产生的。我们合成了一系列用不同醇取代正丁基酯的膜衍生物，发现这些衍生物的逆转活性与醇的碳数有关。从海绵中分离出一种新型的多羟基化醋酸甾醇A作为耐多药的逆转剂。Agosterol A在3μM浓度下完全逆转了KB-C2细胞对秋几丁碱的抗性和KB-CV60细胞对长春新碱的抗性。Agosterol A使KB- c2和KB- cv60细胞长春新碱的积累恢复到3 μM浓度下与亲本KB- 3-1细胞相当的水平。

项目成果

N. Murakami: "Total synthesis of callystatin A, a potent cytotoxic polyketide from the marine sponge, Callyspongia truncata"Tetrahedron Lett.. 39. 23498-2352 (1998)

N. Murakami：“callystatin A 的全合成，一种来自海洋海绵 Callyspongia truncata 的有效细胞毒性聚酮化合物”Tetrahedron Lett.. 39. 23498-2352 (1998)

S.Aoki: "Agostereol A, a novel polyhydroxylated sterol acetate reversing multidrug resistance from a marine sponge of spongia sp"Tetrahedron Letter. 39. 6303-6306 (1998)

S.Aoki：“Agostereol A，一种新型多羟基化甾醇乙酸酯，可逆转海绵的多药耐药性”四面体信件。

S.Aoki: "Agosterol A, a novel polyhydroxylated sterol acetate rerersing multidrug resistance from marine sponge of spongia SP" Tetrahedron Lett. 39. 6303-6306 (1998)

S.Aoki：“Agosterol A，一种新型多羟基化甾醇乙酸酯，可逆转海绵 SP 的多药耐药性”四面体 Lett。