摘要

目的：买朱尼吾西拜出自《卡拉巴丁艾再木》（公元1785-1875年，穆哈穆德·艾扎木汗），为古代经典名方目录（第二批）入选品种。处方由欧菝葜根、地锦草、诃子、番泻叶等共10味药材组成，具有清除异常脾液质和异常黏液质，治疗尿路炎症的作用。本课题按照中药注册分类中“3.1类按古代经典名方目录管理的中药复方制剂”的相关要求，开展买朱尼吾西拜物质基准研究，包括药材质量研究、饮片炮制研究、制备工艺研究、质量标准建立和药效学研究。旨在将其开发成一个独具特色的治疗尿路感染的经典名方，为维药经典名方买朱尼吾西拜的注册申报建立基础。

方法：（1）药材研究：包括关键信息考察、药材质量检查、药材质量提升等。依据2020版《中国药典》与《卫生部部颁标准》等标准，对买朱尼吾西拜组方中收集到的所有批次药材进行质量分析。在原有质量标准上，结合最新文献资料，开展药材标准提升研究，增加单/多成分含量测定、薄层鉴别、灰分、浸出物等质量控制指标，并根据多批次的研究结果制定企业内控标准。（2）毛诃子炒制工艺研究：采用单因素实验法，监测毛诃子中主要成分总皂苷的含量变化，考察确定炒制温度、炒制时间、料液比等关键参数，建立炮制工艺并进行验证，同时进行性状、鉴别、检查、含量测定等多项目研究，建立饮片质量标准。（3）物质基准样品制备研究：包括粉碎工艺与混合工艺。比较单独粉碎方式和串碾粉碎对中间体粉末出粉率、粉体粒径等指标的影响，确定合适的粉碎方式。比较不同混合次数下中间体粉末中安石榴苷、鞣花酸的含量并计算RSD值，选出最佳混合工艺。结合粉碎与混合工艺对十批次的物质基准样品进行质量研究。（4）药效学研究：包括体外抑菌活性研究及体内UTI尿路感染动物实验，采用琼脂倍比稀释法测定买朱尼吾西拜全方及欧菝葜根、地锦草、番泻叶对三种尿道感染菌的MIC值。建立大肠埃希菌急性尿路感染小鼠模型，考察不同剂量买朱尼吾西拜对小鼠一般状态、尿细菌培养的影响，评价其治疗尿路感染的作用。

结果：（1）药材质量检查：共收集到162批药材，经检查155批药材合格，7批次不合格。其中2批毛诃子，2批诃子性状外观不合格，2批地锦草灰分不合格，1批余甘子含量不合格。（2）毛诃子炒制工艺：炒制30 min后，毛诃子黄度值、三种鞣质类成分含量最高，总皂苷含量变化趋于平缓，不再下降。（3）物质基准样品制备：全药材1/6处方量粉碎20 min并混合10次为最佳制备工艺。以最佳工艺进行十批次的物质基准样品研究，验证后工艺稳定可行。（4）药效学结果：买朱尼吾西拜对三种常见尿道菌抑菌效果明显，尿路感染UTI动物模型买朱尼吾西拜中、高剂量组对小鼠尿路感染具有一定的治疗效果。

结论：买朱尼吾西拜组方药材合格率高；毛诃子经炮制后总皂苷含量下降，药效成分增加。炮制工艺、物质基准样品研究、经验证后稳定；买朱尼吾西拜抑菌效果明显，对UTI尿路感染疗效显著。可为买朱尼吾西拜后续开发研究提供基础。

Abstract

Object: Maizhuniwuxibai is from Kalabadin Aizamu (1785-1875 A.D., Muhammad Aizamu Khan), an entry in the Catalog of Ancient Classical Formulas (second batch). The prescription consists of a total of 10 herbs, including Sarsaparilla Ovata Root, Humifuse Euphorbia Herb, Chebulin, and Senna Leaf, which have the effect of removing abnormal splenic and abnormal mucus matter and treating inflammation of the urinary tract. According to the requirements of "Class 3.1 Compound Preparations of Traditional Chinese Medicines Managed According to the Catalog of Ancient Classical Formulas" in the Classification of Traditional Chinese Medicines Registration, this project is to carry out the research on the material baseline of Buizhu Nyusibai, which includes the quality of medicinal materials, concoctions of drinking tablets, preparation process, establishment of quality standards, and pharmacodynamic studies. The aim is to develop it into a unique classic formula for treating urinary tract infections, and to establish a foundation for the registration of Maizhuniwuxibai, a classic prescription of Uyghur medicine.

Methods: (1) Research on herbs: including key information examination, quality check of herbs, and quality improvement of herbs. Based on the 2020 edition of the Chinese Pharmacopoeia and the Ministry of Health Standards and other standards, we analyzed the quality of all batches of medicinal herbs collected in the Maizhuniwuxibai Formulas. On the basis of the original quality standards, combining with the latest literature, we carried out research on the standard improvement of the herbs, added quality control indexes such as single/multiple ingredient content determination, thin-layer identification, ash, leachate, etc., and formulated the internal control standards of the enterprise based on the results of the research on multiple batches.(2) Study on the frying process of Quixote: A single-factor experimental method was used to monitor the changes in the content of total saponin, the main component in Quixote, to determine the frying temperature, frying time, material-liquid ratio and other key parameters, to establish and validate the concocting process, and at the same time to carry out a multi-factor study on the traits, identification, inspection, content determination and other research projects, and to establish a quality standard for the drinking tablets.(3) Substance reference sample preparation study: including pulverization and mixing processes. Compare the effects of individual pulverization and tandem pulverization on the powder yield and particle size of the intermediate powder, and determine the appropriate pulverization method. Compare the content of andrographolide and ellagic acid in the intermediate powder under different mixing times and calculate the RSD value to select the best mixing process. The quality of ten batches of reference samples of the substance was studied by combining the pulverization and mixing processes.(4) Pharmacodynamics research: including in vitro bacteriostatic activity research and in vivo UTI urinary tract infection animal experiments, using agar doubled dilution method to determine the MIC value of the whole formula of Maizhuniwuxibai and Sarsaparilla Roots, Ground Jasmine Herb, Senna Leaf against three kinds of urinary tract infection bacteria. A mouse model of acute urinary tract infection by Escherichia coli was established to investigate the effects of different doses of Maizhuniwuxibai on the general state and urine bacterial culture of mice, and to evaluate its therapeutic effects on urinary tract infection.

Results:(1) Quality inspection of herbs: 155 batches of herbs were qualified after quality inspection, 2 batches of Hairy Chebula, 2 batches of Chebula failed in appearance of traits, 2 batches of Dicentralea failed in ash content, and 1 batch of Eugenol failed in content. (2) A one-way examination was conducted on the concoction of Hairy Chebula, examining the time of concoction, etc., and the content of total saponins as well as three kinds of tannin-like components were determined, as well as the determination of Escherichia coli MIC before and after concoction. (3) Summarize the crushing process according to the crushing characteristics of each medicinal flavor in the prescription of Maizhuniwuxibai, formulate the pre-treatment method of its medicinal herbs, and select the crushing of the whole medicinal herbs in 1/6 prescription quantity for 20 min according to the comprehensive consideration of the rate of powder output and fluidity; then compare the RSD values of the content of aniline and ellagic acid in the powder under different mixing times, and find that it is optimal to mix it for 10 times. The optimal process for the material reference sample was determined and the quality standard of the material reference sample was established.(4) Pharmacodynamic results: Maizhuniwuxibai has obvious bacteriostatic effect on three common urogenital bacteria, and Maizhuniwuxibai in the middle and high dose groups of UTI animal model has certain therapeutic effect on urinary tract infection in mice.

Conclusion:Herbs of Maizhuniwuxibai grouping were qualified; the total saponin content of Maohuaxia decreased and the medicinal components increased after the concoction. The concoction process, material benchmark sample research, and validation were stabilized; Maizhuniwuxibai has obvious antibacterial effect and significant efficacy against UTI urinary tract infection. It can provide the basis for the subsequent development research of Maizhuniwuxibai.

[1]顾思浩,张薇,王冰,等. 经典名方苓桂术甘汤历史沿革[J]. 中国医药导报, 2020, 17（21）: 145-148.

[2]翟艳敏. 经典名方现代研发关键技术[D]. 江苏: 江苏大学, 2021.

[3]刘艳, 章军, 杨林勇, 等. 经典名方物质基准研制策略及关键问题分析[J]. 中国实验方剂学杂志, 2020, 26（01）: 1-9.

[4]陈畅, 程锦堂, 刘安. 经典名方研发策略[J]. 中国中药杂志, 2017, 42（09）: 1814-1818.

[5]张鹏, 刘安, 邬兰, 等. 经典名方概述与开发探讨[J]. 中国实验方剂学杂志, 2020, 26（15）: 183-194.

[6]周菲, 林美斯, 王琳, 等. 经典名方百合地黄汤物质基准制备及过程质量控制研究[J]. 中草药, 2019, 50（16）: 3824-3832.

[7]高乐, 杨梦婷, 冯玉, 等. 经典名方旋覆代赭汤物质基准的质量研究[J]. 中药材, 2022（05）: 1178-1184.

[8]陈士林, 刘昌孝, 张铁军, 等. 基于中药质量标志物和传统用法的中药饮片标准汤剂传承发展研究思路与建议[J]. 中草药, 2019, 50（19）: 4519-4528.

[9]王饶琼, 杨思进, 谢春光, 等. 清肺排毒汤治疗新型冠状病毒肺炎的临床疗效观察[J]. 中药药理与临床, 2020, 36（01）: 13-18.

[10]王檀, 仕丽, 陈亦洋, 等. 新型冠状病毒肺炎50例中医临床疗效分析[J]. 吉林中医药, 2020, 40（03）: 281-285.

[11]赵永旺, 柏莹, 刘峥嵘, 等. 日本汉方医药学发展历程对我国中医药学发展的启示[J]. 湖南中医药大学学报, 2018, 38（05）: 601-604.

[12]赵建, 张俊博. 浅论中医学与日本汉方医学[J]. 内蒙古中医药, 2012, 31（15）: 121.

[13]于翠婷, 田侃, 住田尚之, 等. 日本汉方制剂的发展现状及其经验启示[J]. 中草药, 2018, 49（02）: 494-498.

[14]肖月园, 杨志波. 日本汉方医学发展带来的启示[J]. 中国中西医结合皮肤性病学杂志, 2018, 17（06）: 554-558.

[15]赵晓晖, 马维骐. 日本汉方腹诊发展概况[J]. 国医论坛, 2018, 33（05）: 14-16.

[16]王欢, 朱莹. 经方在中国传统医学和日本汉方医学的应用现状[J]. 中草药, 2019, 50（15）: 3714-3719.

[17]杨平, 林丹, 宋菊, 等. 日本汉方制剂及其特点与中药新药研究的思考[J]. 中草药, 2018, 49（09）: 1985-1989.

[18]高树葳, 邱智东, 赵燕, 等. 基于古代经典名方的中药新产品研究进展[J]. 长春中医药大学学报, 2021, 37（03）: 674-677.

[19]艾买提江·阿衣甫别克, 买买提·努尔艾合买提, 买买提江·阿布都瓦克. 维药“祖发”的本草考证[J]. 中国民族民间医药, 2021, 30（16）: 29-35.

[20]孙嘉磊, 阿依佐合热, 王海生, 等. 新疆维吾尔自治区常用维药、药源植物调查[J]. 时珍国医国药, 2018, 29（08）: 1980-1982.

[21]毛鑫, 姚荣妹, 徐玉辉, 等. 急性尿路感染大鼠模型的建立及在中药药效评价中的应用[J]. 中药药理与临床, 2019, 35（03）: 177-180.

[22]赵秋华, 刘秀蜀, 林乐欣. 阿莫西林克拉维酸钾治疗小儿尿路感染的临床效果[J]. 中国当代医药, 2019, 26（05）: 120-122.

[23]余明主, 郑慧杰, 曹卫霞, 等. 银花泌炎灵片联合左氧氟沙星治疗急性尿路感染的临床观察[J]. 中国医师杂志, 2019, 21（10）: 1571-1574.

[24]陈小丽, 白燕. 中医药治疗淋证的临床研究进展[J]. 内蒙古医学杂志, 2022, 54（04）: 460-462.

[25]胡献国. 淋病的中医治疗[J]. 家庭医学, 2021（01）: 54.

[26]赵俐超, 赵菁莉. 黄文政治疗下尿路感染经验[J]. 湖南中医杂志, 2021, 37（09）: 36-38.

[27]郭秀英. 尿路感染的病因、发病机理与治疗[J]. 基层医学论坛, 2006（12）: 560-561.

[28]王立平. 扶正清热利湿方治疗泛耐药大肠埃希氏菌尿路感染小鼠模型实验研究[D]. 重庆: 西南大学, 2013.

[29]陈文静, 梁文仪, 李师, 等. 基于 HPLC-MSn 和化学计量学的指纹图谱在毛诃子鞣质部位质量评价中的应用 [J]. 世界科学技术—中医药现代化, 2017, 19（7）: 1228- 1233.

[30]崔雨,董婉绒,刘恬恬等. 毛诃子化学成分及药理作用研究进展 [J]. 中国现代中药, 2023, 25 （01）: 216-226.

[31]邓星,罗莉娅,苟立平等. 藏药毛诃子的质量标准提高研究 [J]. 中南药学, 2018, 16 （01）: 117-120.

[32]艾莉,陈君程,张继良等. 基于色彩色差计的川楝子炮制前后含量和颜色变化 [J]. 成都中医药大学学报, 2011, 34 （04）: 81-83.

[33]盛洁静,杨硕. 大黄炮制前后体外抗菌活性比较 [J]. 哈尔滨商业大学学报（自然科学版）, 2019, 35 （06）: 643-644+650.

[34]周坤,简平,梁文仪等. 基于UPLC-Q-Exactive Orbitrap-MS分析藏药诃子与毛诃子化学成分 [J]. 质谱学报, 2020, 41 （03）: 254-267.

[35]李雪冬,罗晓敏,马筝等. 毛诃子化学成分和药理作用的研究进展及其质量标志物（Q-Marker）预测分析 [J]. 中草药, 2023, 54 （03）: 976-990.

[36]李师. 藏药毛诃子化学成分及体外抗癌活性的研究[D]. 北京中医药大学, 2018.

[37]董宇,姜鑫,刘静等. 分光光度法测定宁神颗粒中总皂苷含量 [J]. 食品安全质量检测学报, 2018, 9 （19）: 5208-5211.

[38]欧亮苗,阿来·赛坎,李柯翱等. HPLC法同时测定维药没食子中没食子酸、没食子酸甲酯和鞣花酸的含量 [J]. 国际药学研究杂志, 2015, 42 （05）: 637-641.

[39]黄宽,付鹏,林艾和等. HPLC法同时测定毛诃子中5种鞣质类成分的含量 [J]. 中国药师, 2021, 24 （03）: 607-609.

[40]陈文静,梁文仪,李师等. 基于指纹图谱分析和多成分同时定量的藏药毛诃子质量评价研究 [J]. 中草药, 2017, 48 （06）: 1210-1215.

[41]李秀宇,马芹. 水龙骨中没食子酸与鞣花酸的含量测定 [J]. 化学研究, 2023, 34 （01）: 14-19.

[42]Yadava R N. A new cardenolide from the seeds of Terminalia bellerica [J]. Fitoterapia, 2001, 72（3）: 310-312.

[43]王舒. 药用植物毛诃子研究进展 [J]. 安徽农业科学, 2015, 43 （05）: 65-66.

[44]喻文丽,李佩琪,秦新政等. 不同粉碎工艺对天山雪耳营养成分溶出率及化学成分的影响 [J]. 食品与发酵工业, 2020, 46 （18）: 122-127.

[45]胡慧芬. 姜厚朴均匀化饮片的工艺研究[D]. 湖北: 湖北中医药大学, 2015.

[46]蛋鸡饲料的最佳粉碎粒度 [J]. 家禽科学, 2018, （09）: 7.

[47]王红英,陈计远,金楠等. 饲料粉碎工艺高效利用技术研究 [J].饲料工业,2020, 41 （13）: 1-8.

[48]钟禹文.“扶正清热通淋”汤通过调节NLRP3炎症小体通路对大鼠复杂性尿路感染作用的研究[D]. 四川: 西南医科大学, 2022.

[49]贺天文. 新型大鼠尿路感染模型的建立与中药方剂对产ESBLs大肠埃希菌致大鼠复杂性尿路感染作用的研究[D]. 四川: 西南医科大学, 2019.

[50]石明.小鼠急性细菌性膀胱炎模型的建立及透明质酸钠膀胱灌注治疗小鼠膀胱炎的实验研究[D]. 湖北: 华中科技大学, 2013.

[51]杜婉迎.含色氨酸抗菌肽对铜绿假单胞菌滞留菌的作用研究[D]. 辽宁: 辽宁师范大学, 2022.

[52]苏佳纯,杨帆. 头孢他啶-阿维巴坦在铜绿假单胞菌感染中的应用价值 [J]. 中国感染与化疗杂志, 2023, 23 （02）: 237-242.