基于循证医学理念和血清药物学方法的蒙医瘟疫热病经典方剂"查干汤"抗流感药效物质基础及其代谢规律研究

Chagantang decoction, also known as Siweitumuxiangsan, is used as a basic traditional Mongolian medicine formula for seasonal febrile disease such as flu and other relevant diseases, and could constitute more than 60 other compound prescriptions after adding some other herbal medicines according to the diagnosis. Chagantang has been included in Chinese Pharmacopoeia as first ethnic drug since 1977. Literature survey showed that Chagantang decoction and its component herbs contain chemical constituents with antipyretic, analgesic, anti-inflammatory, anti-bacteria, and immune enhancement activities, which provided certain scientific evidence for the traditional usage of Chigantang as a influenza treatment. However, most of these studies still remain in the stage of bioactivity-guided isolation in vitro. In this study, literatures about clinical research of Chagantang on influenza will be systematically analysed and evaluated based on theory of traditional Mongolian medicine and evidence-based medicine, and the clinical indications and pharmacodynamics index of Chagantang will be selected accordingly; constituents absorbed into blood of experimental animals will be analysed and characterized by the method of serum pharmacochemistry; the influence of constituents which absorbed into blood on the immunity of experimental animals will be measured by serum pharmacological method; the pharmacokinetic of the main active constituents will be studied. The main purpose of the study is to clarify the material therapeutic basis of Chagantang for influenza, and to improve the quality standard of the compound, and to illustrate the pharmacokinetic of the main active constituents, and accordingly, to lay the foundation for the development of an innovative drug for influenza based on secondary development of Chagantang.

"查干汤"又名"四味土木香散",为蒙医治疗流感等瘟疫热病基础方,经辩证加味可形成60多方,1977年作为首批民族用药载入《中国药典》。研究表明,"查干汤"及其组方药材含解热、镇痛、抗炎、抗菌、增强免疫等活性成分，初步阐释了该方用于流感治疗的科学性,但相关研究尚停留在体外分离追踪水平。本课题基于蒙医药理论和循证医学理念系统评价"查干汤"抗流感临床研究文献，初步明确其临床定位和药效指标；采用血清药物化学方法对实验动物血中移行成分进行分析和表征；应用血清药理学方法考察血中移行成分对实验动物免疫功能的影响；对筛选出的主要药效成分进行药代动力学研究。从而明确"查干汤"抗流感的药效物质基础,完善其质量评价体系，阐明方剂主要药效成分的体内药动学过程。为开发研制基于经典蒙药方剂"查干汤"、临床疗效确切、药效物质基础和作用机理较为明确,量效、时效关系较为清楚，安全有效、质量可控的抗流感创新蒙药奠定基础。

基于循证医学理念和血清药物学方法，对蒙医瘟疫热病经典方剂"查干汤"抗流感药效物质基础及其代谢规律进行研究。通过文献整理和临床应用调研，明确其临床定位和药效指标，本品蒙医临床用于治疗病毒性感冒早期、带状疱疹、过敏等，首次提出本品药效指标应与免疫系统作用相关。.药效物质基础研究：⑴化学成分研究，从土木香中分离并鉴定了22 个单体化合物，3个为新化合物（4β,5-dihydroxy-eudesma-11(13)-dien-12,8β-olide、5,6α-dihydroxy-eudesma- 11(13)-dien-12,8β-olide、1(10),4(5),11(13)- germacra-trien-12,8β-olide）；从悬钩子木中分离鉴定了17个化合物，3个为新化合物（1α,2β,3β,19α-四羟基-乌苏-12-烯-28-酸-β-D-吡喃葡萄糖苷、1,6-二咖啡酰基-葡萄糖、悬钩子木质素A）；首次从悬钩子木中分离纯化得到2种天然均一多糖RSP1-1和RSP1-2。⑵体外化学成分析与质量评价，首次建立了悬钩子木药材中表儿茶素的含量测定方法、HPLC指纹图谱、HPLC-MS/MS分析鉴定方法。⑶血中移行成分的分析与表征，建立了高效液相色谱串联四极杆-静电场轨道阱高分辨质谱法（HPLC-Q-Exactive MS），体外共鉴定110个化合物，12个源于土木香，61个源于苦参，30个源于悬钩子木，7个源于山奈；对体内代谢物进行鉴定，共鉴定63个代谢物，其中包括40个原型产物和23个代谢产物。.对机体免疫功能的影响：⑴采用CCK-8法测定本品对小鼠脾淋巴细胞的增殖作用，结果本品及其组方药材的水提物均能不同程度促进脾淋巴细胞增殖，并通过分泌白细胞介素-2和干扰素-γ发挥免疫调节作用。⑵分离得到的悬钩子木多糖RSP1-1和RSP1-2均能不同程度促进脾淋巴细胞的增殖，促进淋巴细胞分泌IL-2、IFN-γ和TNF-α，呈现一定的免疫调节活性。⑶建立小鼠环磷酰胺免疫抑制模型，灌胃给予本品水提物冻干粉，可提高脏器指数，并对血液学指标具有明显的调节作用。⑷土木香中分离得到的倍半萜内酯类化合物，对5种人体癌细胞系均有不同程度的细胞毒活性，通过PI3K-AKT-mTOR和/或RAS-RAF-MEK信号通路抑制肿瘤细胞生长。.开发了安全有效、质量可控的抗流感创新蒙药“四味土木香。