SEARCH FOR METABOLITES INFLUENCING CELLAR FUNCTION FROM AQUATIC INVERTEBRATES AND ANLYSIS OF THEIR MODES OF ACTION.

从日本不同海岸采集的许多海洋无脊椎动物，包括海绵、软珊瑚和被毛类，已经被测试了对P388白血病细胞的细胞毒性，对大鼠神经母细胞3Y1的形态变化诱导活性，对丝氨酸蛋白酶、半胱氨酸蛋白酶、金属蛋白酶和糖苷酶的抑制活性。从有希望的样品中分离和结构鉴定活性成分，发现了71个新化合物，其中包括钙调素抑制生物碱。对新的毛盏花素衍生物和那些从毛盏花素A化学衍生出来的生物活性的研究导致了对毛盏花素与蛋白磷酸酶相互作用的重要见解。对获得的选择性化合物的作用模式进行了详细的分析，得出了几个重要的发现。Mycalolide/kabiramides与肌动蛋白的结合部位不同于细胞松弛素D和牛膝菌素A。双酮内酯A以1：2的摩尔比与G-肌动蛋白结合，以抑制肌动蛋白聚合。Stelletamide A及相关生物碱可通过抑制钙调蛋白而抑制肌球蛋白轻链的磷酸化，从而引起平滑肌收缩。为了寻找影响棘皮动物胚胎发育的代谢物，从海绵中分离出了几种活性物质，包括阿可瑞林A和棒状棕榈酸。Jaspisin被发现选择性地抑制海胆的一种孵化酶，一种基质内肽酶。最后，一个简单的系统，通过使用抗体来区分ANO39(海胆卵的一种内质网蛋白)中的特定磷酸化氨基酸来检测蛋白磷酸酶的抑制物，如冈田酸和Calyculin A。

A number of marine invertebrates including sponges, soft corals, and tunicates collected from various Japanese coasts has been tested for cytotoxicity against P388 leukemia cells, morphological changes-inducing activity in rat neuroblast 3Y1, inhititory activity against serine proteases, cystein proteases, metalloproteinases, and glycosidases. Isolation and structure elucidation of active constituents from promising specimens resulted in discovery of 71 new compounds, among which calmodulin-inhibiting alkaloids were included. Biological activity studies of new calyculin derivatives and those derived chemically from calyculin A led to important insights into the interaction of calyculins with protein phosphatases.Detailed analyses of modes of action of selective compounds obtained led to several inportant findings. Mycalolides/kabiramides bind to a different site of actin from those of cytochalasin D and latrunculin A.Bistheonellide A bind to G-actin in a molar ration of 1 : 2 to inhibit actin polymerization. Stellettamide A and related alkaloids induce contraction of smooth muscles which was found to be due to inhibition of phosphorylation of myosin light chain by inhibiting calmodulin. It was also found that discodermin A forms pores in biomembranes and that theonellamides A and F interact with dehydrogenases.In search for metabolites affecting development of echinoderm embryos, several active substances were isolated from sponges including ancorinoside A and rhopaloic acids. Jaspisin was found to inhibit selectively a hatching enzyme of sea urchin, a matrix endopeptidase. Finally, a simple system to detect inhibitors of protein phosphatases such as okadaic acid and calyculin A by using antibodies which differentiate specific phosphorylated amino acids in ANO39, an endoplasmic reticulum protein of sea urchin eggs.