药学学报, 2019, 54(12): 2195-2203
引用本文:
冯碧, 曹波, 谭鹏, 宋娇, 张海珠, 许洪, 杜晓娟, 牛明, 许润春, 杨明, 韩丽, 张定堃. 化学指纹图谱与活血、抗炎生物效价在小金丸整体质量评价中的适宜性对比研究[J]. 药学学报, 2019, 54(12): 2195-2203.
FENG Bi, CAO Bo, TAN Peng, SONG Jiao, ZHANG Hai-zhu, XU Hong, DU Xiao-juan, NIU Ming, XU Run-chun, YANG Ming, HAN Li, ZHANG Ding-kun. Comparative study on the suitability of chemical fingerprint and anti-platelet aggregation/anti-inflammatory biopotency in the overall quality evaluation of Xiaojin pills[J]. Acta Pharmaceutica Sinica, 2019, 54(12): 2195-2203.

化学指纹图谱与活血、抗炎生物效价在小金丸整体质量评价中的适宜性对比研究
冯碧1, 曹波1, 谭鹏2, 宋娇1, 张海珠3, 许洪4, 杜晓娟4, 牛明5, 许润春1, 杨明6, 韩丽1, 张定堃1
1. 成都中医药大学药学院, 西南特色中药资源国家重点实验室, 四川 成都 611137;
2. 四川省中医药科学院, 国家中医药管理局中药质量生物评价重点研究室, 四川 成都 610041;
3. 大理大学药学与化学学院, 云南 大理 671000;
4. 成都永康制药有限公司, 四川 成都 611137;
5. 中国人民解放军总医院第五医学中心, 全军中医药研究所, 北京 100039;
6. 江西中医药大学, 江西 南昌 330004
摘要:
以中医临床治疗乳腺增生的首选中成药小金丸为例,对比生物活性检定与化学指纹图谱在小金丸整体质量评价中的适宜性与科学性。测定小金丸抗血小板聚集生物效价与环氧化酶2(cyclooxygenase-2,COX-2)抑制率,建立小金丸脂溶性和水溶性双部位指纹图谱,对A、B 2个厂家16个批次市售小金丸进行指纹图谱分析,对比研究小金丸在生物活性和化学分析层面的一致性。本研究动物实验方案已获成都中医药大学附属医院实验动物伦理学委员会批准(批准号:2018BL-002)。结果表明,16个批次小金丸的抗血小板聚集生物效价在0.712~1.278 U·mg-1,相对标准偏差(RSD)为15.4%;COX-2抑制率在52.07%~68.95%,RSD为8.91%,结果显示生物效应差异较小,生物活性一致性较好。16个批次小金丸指纹图谱相似度差,近一半样品相似度小于0.9,共有峰峰面积在各样品中RSD达32.74%~165.37%,化学一致性差。为探究小金丸生物活性一致性好而化学一致性差的原因,对指纹图谱色谱进行多元统计分析,筛选影响相似度的主要色谱峰,并进行色谱峰药材归属指认。结果显示各批次小金丸的相似度主要受脂溶性指纹图谱中面积较大的17、18、20、23、27号色谱峰,水溶性部分30号峰的影响,经鉴定这些色谱峰属于枫香脂和当归。进一步研究不同浓度枫香脂、当归抗血小板聚集与抑制COX-2活性的作用,结果表明在小金丸组方比例下,枫香脂、当归几乎没有抗血小板聚集和抑制COX-2活性的作用。本研究发现化学指纹图谱用于小金丸质量一致性评价存在一定的局限性,部分丰度高、光谱响应强、活性弱的色谱峰成为影响评价结果的主要因素,选择反映临床功效的生物评价方法能很好弥补化学评价方法的不足,为复杂中成药的整体质量评价提供新思路与方法。
关键词:    小金丸      质量评价      指纹图谱      抗血小板聚集      COX-2抑制率     
Comparative study on the suitability of chemical fingerprint and anti-platelet aggregation/anti-inflammatory biopotency in the overall quality evaluation of Xiaojin pills
FENG Bi1, CAO Bo1, TAN Peng2, SONG Jiao1, ZHANG Hai-zhu3, XU Hong4, DU Xiao-juan4, NIU Ming5, XU Run-chun1, YANG Ming6, HAN Li1, ZHANG Ding-kun1
1. State Key Laboratory of Characteristic Chinese Medicine Resources in Southwest China, College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China;
2. State Key Laboratory of Quality Evaluation of Traditional Chinese Medicine, Sichuan Academy of Traditional Chinese Medicine, Chengdu 610041, China;
3. Department of Pharmacy and Chemistry, Dali University, Dali 671000, China;
4. Chengdu Yongkang Pharmaceutical Co., Ltd., Chengdu 611137, China;
5. China Military Institute of Chinese Medicine, the Fifth Medical Centre of Chinese People's Liberation Army General Hospital, Beijing 100039, China;
6. Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China
Abstract:
Xiaojin pills, the first choice for clinical treatment of breast hyperplasia, were selected to explore the suitability of a bioactivity assay with chemical fingerprinting for the development of an overall quality evaluation assay. The liposoluble and water-soluble fraction fingerprints of Xiaojin pills were established. The ability to inhibit platelet aggregation and the rate of inhibition of cyclooxygenase-2 (COX-2) for 16 batches of Xiaojin pills from several manufacturers was analyzed; the chemical fingerprints of these samples were correlated with the bioactivity and chemical analysis. The animal protocol was approved by the Committee on the Ethics of Animal Experiments of Affiliated Hospital of Chengdu University of Traditional Chinese Medicine Approval, ID:2018BL-002. Results showed that the antiplatelet aggregation activity of 16 batches was 0.712-1.278 U·mg-1, with a relative standard deviation (RSD) of 15.4%. COX-2 inhibition was 52.07%-68.95% and the RSD was 8.91%. The results showed that there was little difference in the biological effects of these samples. However, the chemical fingerprint consistency of these 16 batches of Xiaojin pills was poor, and the similarity of nearly half of the samples was less than 0.9. The total peak area of Xiaojin pills was 32.74%-165.37% across samples, showing very poor chemical consistency. In order to explore the reasons for the poor chemical consistency despite good consistency in the biological assays, the fingerprint chromatogram was analyzed by multivariate statistical analysis. The main chromatographic peaks were identified. The results showed that the similarity of Xiaojin pills was mainly determined by the prominent chromatographic peaks 17, 18, 20, 23 and 27 in the liposoluble fingerprints, which were identified from Liquidambaris resina and Angelica sinensis Radix. However, Liquidambaris resina and Angelicae sinensis Radix had almost no anti-platelet aggregation activity or COX-2 inhibitory effect at the normal prescription ratio. As a result, the ability to utilize chemical fingerprints to evaluate the quality consistency of Xiaojin pills is limited. The selection of biological evaluation methods that reflect clinical efficacy could make up for the shortcomings of chemical evaluation methods for quality assessment, and provide new ideas and methods for the overall quality evaluation of complex Chinese patent medicines.
Key words:    Xiaojin pill    quality evaluation    fingerprint    antiplatelet aggregation    COX-2 inhibition rate   
收稿日期: 2019-08-20
DOI: 10.16438/j.0513-4870.2019-0667
基金项目: 国家自然科学基金面上项目(81873232,81573676);成都中医药大学中药学学科特色创新科研团队(CXTD2018006);四川省科技厅应用基础研究项目(2019YJ0640).
通讯作者: 韩丽,E-mail:hanliyx@163.com;张定堃,E-mail:zhangdingkun@cdutcm.edu.cn
Email: hanliyx@163.com;zhangdingkun@cdutcm.edu.cn
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参考文献:
[1] Xiao XH, Zhang DK, Wang JB, et al. Comprehensive quantitative evaluation and control system for traditional Chinese medicine quality-power pyramid of standard evaluation[J]. China J Chin Mater Med (中国中药杂志), 2015, 40:7-12.
[2] Tanaka R, Takahashi N, Nakamura Y, et al. In-line and real-time monitoring of resonant acoustic mixing by near-infrared spectroscopy combined with chemometric technology for process analytical technology applications in pharmaceutical powder blending systems[J]. Anal Sci, 2017, 33:41-46.
[3] Chen S, Sun G, Ma D, et al. Quantitative fingerprinting based on the limited-ratio quantified fingerprint method for an overall quality consistency assessment and antioxidant activity determination of Lianqiao Baidu pills using HPLC with a diode array detector combined with chemometric methods[J]. J Sep Sci, 2018, 41:548-559.
[4] Yi LZ, Wu H, Liang YZ. Chromatographic fingerprint and quality control of traditional Chinese medicines[J]. Chin J Chromatogr (色谱), 2008, 26:166-171.
[5] Zhao CP. Overview of traditional Chinese medicine fingerprints[J]. Chin J Trauma Disabil Med (中国伤残医学), 2014, 5:290-291.
[6] Sun TT, Ma XH, Li XX, et al. Discussion on the status quo and development ideas of traditional Chinese medicine bio-potency[J]. Chin Tradit Herb Drugs (中草药), 2017, 48:1906-1911.
[7] Li XJ, Huang YY, Yang Z, et al. Research strategy of biomarkers for quality of traditional Chinese medicine based on effect benchmark[J]. Acta Pharm Sin (药学学报), 2019, 54:34-40.
[8] Sun LK, Liu F, Chen WQ, et al. Meta-analysis-based evaluation of Xiaojin pills in the treatment of breast hyperplasia[J]. Chin J Drug Eval (中国药物评价), 2016, 33:448-451.
[9] Zhang D, Ma R, Lin CX. Clinical observation of Xiaojin pills in the treatment of multiple small nodular goiter[J]. Mod J Integr Tradit Chin West Med (现代中西医结合杂志), 2016, 25:309-311.
[10] Qu JN. 80 cases of benign prostatic hyperplasia treated with Xiaojin pills[J]. World Chin Med (世界中医药), 2008, 3:277.
[11] Luo ZP, Zhou YX. Progress in clinical application of Xiaojin pills[J]. Chin Pract Med (中国实用医药), 2009, 4:227.
[12] Wei ZY, Pan LY, Tan Y. Observation on therapeutic effect of Xiaojin pills on chronic hepatitis B hepatic fibrosis[J]. Chin Pharm (中国药业), 2011, 20:73.
[13] Xiong X, Feng B, Cao B, et al. Study on the modern research of Xiaojin pills and analysis of key programs[J]. China J Chin Mater Med (中国中药杂志), 2018, 43:4801-4807.
[14] Xiong X, Pan Y, Zhang DK, et al. Quantitative determination of HPLC-MS/MS in combination with stoichiometry to study the quality consistency of commercially sold Xiaojin pills[J]. Chin Tradit Herb Drugs (中草药), 2017, 48:2189-2196.
[15] Xiong X, He YN, Bi F, et al. Screening for the anti-inflammation quality markers of Xiaojin pills based on HPLC-MS/MSmethod, COX-2 inhibition test and protein interaction network[J]. Sci Rep, 2018, 8:7454.
[16] Qu XW, Zhang SW, Zhang PH, et al. Xiaojin pills inhibits the expression of COX-2 in prostate tissues of prostatitis pain rats[J]. Nat J Androl, 2008, 14:759-762.
[17] Li HB. Clinical observation of Xiaojin pills combined with Youjiale in the treatment of nodular goiter[J]. Chin Manip Rehabil Ther (按摩与康复医学), 2016, 7:41-42.
[18] An MJ, Cheon JH, Kim SW, et al. Guggulsterone induces apoptosis in colon cancer cells and inhibits tumor growth in murine colorectal cancer xenografs[J]. Cancer Lett, 2009, 279:93-100.
[19] Zhou AG, Ding YX, Guo HY, et al. Experimental investigation on the anti-cancer effect of modifed Xiaojin pills[J]. J Shanghai Second Med Univ (上海第二医科大学学报), 1990, 3:185-188.
[20] Qu XW, Zhang SW, Zhang PH, et al. Effect of Xiaojin pills on COX-2 expression in prostate tissue of rats with prostatitis pain[J]. Nat J Androl (中华男科学杂志), 2008, 8:759-762.
[21] Xu DD, Bao YR, Liu Y, et al. Overview of research on antiplatelet aggregation mechanism of traditional Chinese medicine[J]. Chin J Ethnomed Ethnopharm (中国民族民间医药), 2018, 27:88-91.
[22] Hua DF. A review of existing Chinese patent medicines for adjuvant anti-tuberculosis therapy[J]. J Clin Pulm Med (临床肺科杂志), 2010, 15:1286-1288.
[23] Zhang h, Wang YT, Yan C, et al. Research progress and comparison of pharmacological effects of Angelica and Ligusticum wallichii[J]. Lishizhen Med Mater Med Res (时珍国医国药), 2008, 19:293-295.
[24] Food and Drug Administration. Botanical Drug Development Guidance for Industry. 2016-12-27[2019-08-20]. https://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm458484.pdf.
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