Preparation/production and Utilization Researches of Terpenoidal Plant-growth Stimulants and Other Bioactive Substances with High Bioactivity.

1.利用反序半合成技术制备高生物活性的梭菌素衍生物我们通过合成梭菌素的真正生物合成中间体——梭菌卡-2,10(14)-二烯的19/20-氟衍生物，成功地获得了19/20-氟梭菌素，并将其输入产生梭菌素的真菌中进行代谢转化。我们将这种新工艺命名为“逆序半合成”，因为这种方法与典型的半合成相反，因为纯化学步骤之后是生化转化。在日本发现新的Fusicoccum canker真菌（Phomopsis amygdali Niigata）是日本第一个产生fusicocin的真菌，并选择其Niigata 74菌株作为fusicocin产量最高的菌株之一。该菌株在商业糖豆粕培养基中培养一周产生梭菌素a和3′-去乙酰梭菌素a（各0.4 g/L），培养两周仅产生3′-去乙酰梭菌素a （0.9g/L）。从Niigata菌株中分离出74个新的丝裂菌素P、Q、R、S和3-表丝裂菌素H、Q为极性丝裂菌素，并通过化学和光谱证据对其结构进行了鉴定。镰刀菌素P和R为16- o -甲基镰刀菌素，在脱落酸存在下的种子促萌发试验中表现出较高的活性。Cotylenin A诱导哺乳动物细胞分化活性及其肿瘤细胞生长抑制活性在原代培养中，Cotylenin A诱导人急性髓系白血病细胞分化，并在低浓度下对PC12神经细胞表现出神经突生成（生长）促进活性。子叶列宁A和干扰素α协同诱导人非小细胞肺癌裸鼠细胞凋亡并具有抗肿瘤作用。

1.Preparation of Highly Bioactive Fusicoccin Derivatives by Reverse-order Semi-synthesisWe have succeeded in obtaining 19/20-fluorofusicoccins quite efficiently by synthesizing 19/20-fluoro derivatives of fusicocca-2,10(14)-diene, a genuine biosynthetic intermediate of fusicoccins, and then feeding them into the fusicoccin-producing fungus for metabolic transformations. We named this novel process a "Reverse-order Semi-synthesis", as the approach deviates reversely from typical semi-synthesis insofar as purely chemical steps are followed by biochemical transformations.2.Finding of New Fusicoccin-producing Fungi in Japan and Gram-scale Production of FusicossinsWe found peach Fusicoccum canker fungi (Phomopsis amygdali Niigata) as the first fusicoccin-producing fungi in Japan, and selected its Niigata 74 strain as one of the highest fusicoccin-productive strains. This strain in a commercial sugar-soybean meal medium produced fusicoccin A and 3'-deacetylfusicoccin A (each 0.4 g/L) for its one-week cultivation or only 3'-deacetylfusicoccin A (0.9g/L) for its 2-week cultivation.3.Isolation of New Fusicoccins and Their Seed Germination-stimulating ActivityFrom the strain Niigata 74 new fusicoccins P, Q, R, and S, and 3-epifusicoccins H and Q were isolated as polar fusicoccins, and their structures were elucidated by chemical and spectroscopic evidence. Fusicoccins P and R being 16-O-methylfusicoccins showed the high activity in the seed germination-stimulating test in the presence of abscisic acid.4.Mammal Cell Differentiation-inducing Activity of Cotylenin A and Its Tumor Cell-Growth Suppressive ActivityCotylenin A induced the differentiation of human acute myeloid leukaemia cells in primaiy culture, and showed neurite genaration (outgrowth) promoting activity toward PC12 nerve cells at low concentrations. Cotylenin A and interferon-α cooperatively induced apoptosis and have an antitumor effect on human non-small cell lung carcinoma cells in nude mice.