PRODUCTION OF ARTEMESININ BY CELL CULTURES

通过细胞培养生产青蒿素

• 批准号: 2064765
• 负责人: RAMACHANDRAN M NAIR
• 金额: $ 11.97万
• 依托单位: CITY COLLEGE OF NEW YORK
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 1995-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2064765
• 关键词: antimalarial agents; biotechnology; cell free system; drug design /synthesis /production; medicinal plants; parasitic disease chemotherapy; parasitic diseases; plant extracts; tissue /cell culture

DESCRIPTION (Adapted from the Applicant's Abstract): Artemisinin (Qinghaosu) is a potent antimalarial produced by the Chinese herb, Artemisia annua L. and is particularly effective in the treatment of the parasitic infections caused by chloroquine-resistant Plasmodium species. In clinical studies conducted in the Peoples Republic of China, spanning over a decade, involving thousands of human patients, artemisinin was found to be relatively safe and free of adverse side effects. However, at present, it is available for treatment, only in the Peoples Republic of China where the independent clinical evaluation as a prophylactic, and for mass treatment in other areas where malaria is endemic, is particularly urgent now, because of the steady proliferation of chloroquine-resistant strains. The primary objective of this project is to employ, as complementary, or as an alternative, to whole plant extraction, the following highly effective biotechnological approaches, namely, cell suspension cultures and bioconversion using cell-free enzymes. Dr. Nair and co-workers recently demonstrated in preliminary experiments that cell suspension cultures of A. annua produces artemisinin. They now propose to obtain new cell lines which are better producers of the drug, by varying the growth hormones and other parameters governing callus formation. These cell lines will then be used in suspension cultures under varying conditions to optimize artemisinin production. Finally, pilot plant scale studies will be initiated to determine the economic viability of the process. Intermediates in the biosynthesis of a compound can be less complex and thus more easily accessible by synthesis, and hence, are attractive starting compounds for bioconversion studies. Terpenoids closely related to artemisinin, as well as those of cadinane and germacrane groups, have been postulated as precursors of it. However, the only reported biotransformation using cell-free extracts has been that of arteannuin B. Now, artemesic acid, another metabolite of A. annua, which is more abundant (> 10 times) than artemisinin in some strains, has also been converted to the latter. Parallel to these studies, they will use labeled, basic building blocks to define the biogenetic pathway of this terpenoid and to identify the intermediates. Partially purified and/or immobilized enzymes derived for leaves and cell cultures will be used for bioconversions. The proposed approaches will lead to additional routes for an adequate supply of artemisinin; they also can result in the isolation of more potent derivatives of the drug by its bioconversion by the cell cultures and cell-free extracts.

描述（改编自申请人的摘要）：阿耳马素 （qinghaosu）是一种有效的抗疟药，由中国草药产生 Artemisia Annua L.，在治疗 由氯喹引起的寄生虫感染。 在中华民国进行的临床研究中 十多年来，涉及成千上万的人类患者，发现了阿耳马素 相对安全，没有不利的副作用。 但是，在 目前，只能在人民共和国进行治疗 中国独立的临床评估是一种预防性的，并且 在疟疾流行的其他地区的大规模治疗，尤其是 现在很紧急，因为耐氯喹的稳定扩散 菌株。 该项目的主要目的是雇用 补充，或作为替代方案，与整个植物提取， 遵循高效的生物技术方法，即细胞 使用无细胞酶的悬浮培养物和生物转化。 Nair博士和同事最近在初步实验中证明了 A. Annua的细胞悬浮培养物产生阿秒蛋白。 他们现在 提议获得新的细胞系，该细胞系更好地是该药物的更好生产者 改变愈伤组织的生长激素和其他参数 形成。 然后，这些细胞系将用于悬浮培养 各种条件以优化青蒿素的产生。 最后，飞行员 将开始植物规模研究以确定经济生存能力 过程。 化合物的生物合成中的中间体可能不那么复杂，并且 因此，可以更容易通过合成，因此很有吸引力 启动生物转化研究的化合物。 萜类密切相关 到青蒿素以及cadinane和菌氨基烷基的青蒿素，具有 被认为是它的前体。 但是，唯一的报道 使用无细胞提取物的生物转化是Arteannuin B的。 现在，A。Annua的另一种代谢物artemesic Acid，更丰富 （>> 10次）比某些菌株中的青蒿素也被转换为 后者。 与这些研究平行，他们将使用标记的，基本的 建立块以定义该萜类化合物的生物遗传途径 识别中间体。 部分纯化和/或固定酶 用于叶子和细胞培养物的衍生物将用于生物转化。 提出的方法将导致足够的其他路线 阿秒蛋白的供应；它们还可以导致隔离更有效 该药物通过其生物转化的衍生物通过细胞培养物和 无细胞提取物。