基于药代动力学信息追踪光叶菝葜和小叶菝葜抗痛风性关节炎药效物质及其构效关系的研究

The gouty arthritis is a type of Traditional Chinese medicine "arthralgia" category, and the clinical incidence rate is high,Chemical drugs with toxic side effect such as colchicin, allopurinol and so on are used to treat the disease, and the patients are anguished. There is no ideal curative drug now. Reported in the literature and the results of our research show that the Smilax glabra Roxb and Smilax microphylla have the effect on gouty arthritis. They are widely used in fork, and the effect is significant. The research group has investigated on the chemical constituents of Smilax microphylla systematically. Frty five compounds such as flavonoid and steroid saponin were separated from Smilax macrophylla, and some new compounds were found. This study will be carried out by a four-step "Pharmacodynamic screening- - -Pharmacokinetics process- - -active monomer compounds- - -clearing structure-activity relationship". On the basic of chemical effective constituents separated and identified systematically from Smilax glabra Roxb and Smilax microphylla and based on the constituents absorbed into blood during the pharmacokinetic process, the effective monomer compounds of anti-goutyarthritis will be tracked, and the method of computational chemistry is applied to build pharmacophore. The study will provide lead compound for the research of selecting a new type of anti-gout arthritis drug, also provide important theoretical basis for the design of the plan of more active lead compounds.

痛风性关节炎属于中医痹症范畴，临床发病率高，常用的化学药如秋水仙碱、别嘌呤醇等毒副作用大，患者痛苦，目前尚无理想的治疗药物。 文献报导和本课题组的研究结果表明，光叶菝葜和小叶菝葜都具有抗痛风性关节炎的作用，民间使用广泛，效果显著。课题组前期针对小叶菝葜的化学成分进行了系统的研究，分离得到黄酮类和甾体皂苷类等45个化合物，发现了一些新的化合物。 本研究拟通过"药效筛选- - -药代动力学过程- - -活性单体化合物- - 明晰构效关系"四个步骤，在系统分离鉴定光叶菝葜和小叶菝葜的有效物质基础上，以药代动力学过程中入血移行成分为依据，追踪抗痛风性关节炎的有效单体化合物，并采用计算化学的方法构建药效团。 本研究将为选择新型的抗痛风性关节炎药物研究提供先导化合物，同时也为设计活性更强的先导化合物提供重要理论基础。

1. 首次提取土茯苓的总黄酮，已知成分标示量达到50%以上。50 kg的土茯苓药材70%乙醇回流提取2次，得到的浸膏进行萃取，获得乙酸乙酯部位干浸膏586 g、正丁醇部位干浸膏588 g。进一步考察聚酰胺柱富集土茯苓总黄酮的最优工艺：选用60-100目聚酰胺，最大上样量0.1111 g乙酸乙酯部位/g聚酰胺，上样后静置2 h，先用2 BV水洗脱，2 BV 10%乙醇除杂质，最后50%乙醇洗脱收集4 BV，旋干，得到土茯苓总黄酮富集物，可标识的总黄酮含量约为54.98%，达到国家五类新药标准。.2. 土茯苓总黄酮富集物各给药组均能显著降低高尿酸血症小鼠血尿酸值（p<0.01或p<0.05），高、中剂量组相对模型组的血清尿素氮值有显著下降（p<0.01），低剂量组能有效降低小鼠的血清肌酐值（p<0.01）。土茯苓总黄酮富集物高、中剂量给药组能够降低受试小鼠24h尿液中的尿酸与肌酐值（p<0.01），中剂量组还降低了小鼠肝脏黄嘌呤氧化酶的含量（p<0.05）。.3. 土茯苓总黄酮提取物不同浓度给药组均能减少尿酸钠所致大鼠急性足肿胀大鼠的足肿胀程度（p<0.01），其中中剂量给药组的IL-1β值显著下降(p<0.05)，低剂量组能显著抑制IL-6，TNF-α的产生（p<0.05）。.4. 与正常组比较，模型组小鼠肾脏有机阴离子转运体1（OAT1）和肉碱/有机阳离子转运体（OCTN2）蛋白表达及mRNA表达均下降；与模型组比较，土茯苓总黄酮富集物高剂量组和中剂量组显著上调高尿酸血症小鼠肾脏OAT1和OCTN2蛋白表达水平，土茯苓总黄酮富集物高剂量组显著上调高尿酸血症小鼠肾脏OAT1和OCTN2 mRNA的表达水平。提示土茯苓总黄酮富集物可能通过增加尿酸分泌来促进高尿酸血症小鼠尿酸排泄以降低血清尿酸水平。.5. 正在申请土茯苓总黄酮提取和药效相关专利，为开发成降尿酸中药奠定基础。