Exploration of Thoughts and Methods in Study on Material Base of Traditional Chinese Medicinal Herbs Prescriptions

WORLD SCIENCE AND TECHNOLOGY MODERNIZATION OF TRADITIONAL CHINESE MEDICINE AND MATERIA MEDICA

Volume 11, Issue 4, August 2009 Online English edition of the Chinese language journal Cite this article as: Mode Tradit Chin Med Mater Med, 2009, 11(4): 488–492

REVIEW

Exploration of Thoughts and Methods in Study on Material Base of Traditional Chinese Medicinal Herbs Prescriptions Zhao Xiaoli1, Zhang Xinzhuang1, Di Liuqing1*, Wang Shouchuan2, Cai Baochang3, Du Qiu1, Shan Jinjun1, Bi Xiaolin1, Wei Hongmei1 1

Research Institute of Chinese Medicine and Pharmacology, Nanjing University of Chinese Medicine; Nanjing, Jiangsu 210029, China; Department of Pediatrics, Nanjing University of Chinese Medicine; Nanjing, Jiangsu 210029, China; 3 Jiangsu Key Laboratory of Chinese Medicine Processing, Nanjing University of Chinese Medicine; Nanjing, Jiangsu 210029, China 2

Abstract: The study on the material base of compound preparation lays a foundation for the development of preparation techniques for traditional Chinese medicine (TCM). The study ideas and methods directly influence the rationality and the clinical efficacy of these techniques. This paper gives a general review on the current characteristic of the study ideas and methods, and it also presents some new ideas and methods for the study on the material basis of TCM compound preparation. Key Words: Traditional Chinese herbs prescriptions, material base, thoughts, methods

1

Introduction

Traditional Chinese medicine (TCM) compound preparation is a basic Chinese medicine form accumulated by ancient medical practitioners through long-time medical experiences. TCM compound preparation is a different form from the simple opposite and complementary intervention mold. TCM herbs prescription is under the guidance of TCM theories and syndrome differentiation. The prescription is given based on the cause and the mechanism of the disease, and on the basis of the properties of TCM herbs. The prescription composition follows the rule of “principal-assistant-adjuvant-mediating guide” of TCM theory. Thus, the composition of herbs becomes “an army with strict discipline” so as to achieve comprehensive therapeutic effects. TCM herbs prescription has good therapeutic effects; however, due to the indistinctness of its effective substances, there are still some problems that influence TCM preparation process. In recent years, with the development of subjects, such as cellular biology, molecular biology, computer technology, chromatographic techniques, genomics, and proteomics, and the application of fingerprint, high-throughput screening, and computer-aided technique, in the study of material base of TCM compound preparation, many new thoughts and methods are proposed. This paper gives a review on literature study and the earlier laboratory study, and then it also puts forward a thought in the study of

material basis of TCM compound preparation.

2 Thoughts in study on material base of TCM herbs prescriptions Luo [1] proposed that the material base of TCM herbs prescriptions is composed of general chemical constituents, which are inorganic constituents, organic constituents, and trace elements. The study should also combine chemical constituents with action mechanism and effect of Chinese medicine to explain characteristics of multitargets and synergistic actions of TCM herbs prescription. The study was summarized as “one combination, two basic understandings, three chemical and four pharmacological levels,” which is combined with modern scientific techniques and analytical instruments in the study of material base of TCM herbs prescriptions. Wu [2] suggested that the study methods on material base of TCM herbs prescriptions should be the same as the study on the material base of a single herb. The study should be launched on the basis of TCM syndrome models and evaluated by the therapeutic effects and indications of the compound preparation. Mathematical methods are applied in removing noneffective constituents from the system in order to identify the effective system of the compound preparation. Liu [3] believed that the material base of TCM herbs prescriptions is the effective constituents based on the combination of a disease and its syndromes. The study should be fur-

Received date: 6 August 2009 * Corresponding author. Tel: +86-25-85811071; E-mail: [email protected] Foundation item: Supported by the National High Technology Research and Development Program of China (863 Program) (No.2003AA2Z3247), National Natural Science Foundation of China (30873401), Natural Science Foundation of Jiangsu Province (BK2005150), and Open Funds of Key Subject on TCM Pediatrics, Nanjing University of Chinese Medicine (EZK2009019). Copyright © 2009, World Science and Technology Press. Published by Elsevier BV. All rights reserved.

Zhao Xiaoli et al. / Mode Tradit Chin Med Mater Med, 2009, 11(4): 488–492

ther explored on the establishment of animal, organ, and cell models under the guidance of TCM and modern medical theories, as well as the therapeutic effect of the compound preparation. Thus, the material base of TCM herbs prescriptions will be explained. Ma [4] held that the chemical constituents of TCM herbs prescriptions influence the combination of signal molecules and restore the disordered signal molecules to their balance in the network in order to treat TCM syndrome and diseases. Thus, a new concept of “TCM moleculomics” is proposed in the study on material base of TCM herbs prescriptions. Huang [5], based on the hypothesis of syndrome and treatment pharmacokinetics, suggested that not all chemical constituents in the serum have pharmacological effect and biological activity after oral administration. Only those that are transported to targeting sites of the disease can be referred to as the real effective compositions in the herbal compounds. Therefore, a new hypothesis of tissue pharmacology of prescription was suggested. The above-mentioned thoughts suggested study of material base of TCM herbs prescriptions from different angles. They are reasonable and provide guidance for its study in certain time. However, due to the diversity of chemical constituents and effects, the complex processes of human body, the multilevel interaction between compound preparation and the body, and the limited understanding of TCM theories, currently, it is difficult to fully explain material base of TCM herbs prescriptions.

3 Approaches to the study on material base of medicinal effectiveness in TCM herbs prescripttions According to the thoughts on the above-mentioned researches, research methods applied in the study on material base of TCM herbs prescriptions fall into two categories. The first category is to separate the active chemical constituents with chemical methodologies, and to study the pharmacological effect in order to search for the material base of compound preparations. Methods applied in this category are chemical matteromics and high-throughput screening with multitarget. The second category is to study the material base of compound preparations through samples, such as body fluid and blood. Medicine components are absorbed through body. Study methods used in this category are serum pharmacology, molecular biochromatography, and metabonomics. 3.1 Serum pharmacology and serum pharmacochemistry TCM serum pharmacochemistry, since its concept and detailed rules were suggested by Wang Xijun, has been continuously improved in the study of Chinese herbs. The theory of this method is that most herbs are transformed into complex chemical constituents during herb preparation (e.g.

processing, decocting) and a series of effect after oral administration, such as gastric acid, digestive fluid, intraintestinal enzymes, intestinal flora, liver enzyme, and so on. It is considered that only parts of the compounds are selectively absorbed into the body and be effective on target tissues. Therefore, the hypothesis that chemical constituents in the blood are the real effective substances was suggested [6]. This method is considered as one of the main research methods in the study of material base of TCM herbs prescriptions. Jin [7] studied the oral administration of Gegen Qinlian decoction and each individual herb in the prescription on rats. The comparison of high-performance liquid chromatography (HPLC) fingerprints of plasma samples revealed that 31 constituents enter into the blood. And 15 of them remain as the original constituents contained in Gegen Qinlian decoction, 13 of them are metabolites while three of them are new constituents produced by oral administration of the decoction. The study initially identifies the material base of the compound preparations. Classic prescription [8, 9] Liuwei Dihuang pill was thoroughly studied by serum pharmacochemistry. The material base of kidney tonification was identified and initially separated from the serum. In vitro experiment showed that the effect of morroniside, sweroside, and loganin are the most active constituents and considered as the core factors in kidney tonification. Parts of the material base of compound preparation were explained in the study of this prescription. It has laid a solid foundation for studies in the future. This method combines compound preparations with their action subject in order to remove the distraction of complex physical and chemical properties (e.g. electrolyte, tannin, pH, osmotic pressure, and so on). It narrows the searching range of material base and helps revealing the real effective constituents. This method provides a new method for the secondary development of compound preparation [10]. But there are still some disadvantages in this method at present, such as limitations of detection approach and complexity of endogenous substance. Therefore, not all constituents that enter into the blood can be detected; especially those trace compositions which play an important role in the therapeutic effects [11]. The quality and quantity of constituents absorbed into the blood also change according to the condition of the body. Thus, the therapeutic effects are different. Till date, no standard concerning TCM syndromes are confirmed. So the research results can only be used to show certain aspect of the material base of TCM herbs prescriptions. 3.2 Combinatorial chemical research of TCM and chemical matteromics Enlightened by the theory of “the natural combinatorial chemical libraries,” Yang [12, 13] suggested a new strategy of combinatorial chemical research of TCM in the year of

Zhao Xiaoli et al. / Mode Tradit Chin Med Mater Med, 2009, 11(4): 488–492

1998. This method is based on the mechanism of multitarget of Chinese material medical compounds (CMMC), which are the natural combinatorial chemical libraries. Active compounds are extracted by screening methods according to bioactive target and affinity chromatography from libraries under the guidance of TCM theories. Then TCM chemical constituents are used in different chemical libraries in order to determine their structures and bioactivities. Finally, through analysis and summation, the material base of compound preparation is explained [10]. Combined with theories of systematic biology, Luo [14] proposed “chemical matteromics” and successfully applied it in the study of Qingkailing compound and its global chemome, which was divided into nine subchemomes. Then the matrix of chemical information and the matrix of pharmacologic active information were correlated and a nonlinear composition-effectiveness relation was applied in the identification of screening effective constituents group in TCM. Finally, four subchemomes effective in treating cerebral ischemia injury of Qingkailing are extracted, which are bile acids, baicalin, iridoids from cape jasmine fruit, Concha margaritifera. This method can be used to simplify the diversity of molecules in the prescription in order to develop simplified compound preparation. It emphasizes both the concept of holism and syndrome differentiation. This is another research method for the modernization of TCM. However, as this is just the beginning of this technology, some standard technique and method have not been established. Further research in this system is needed. Besides, the construction of a new prescription should be guided by modern medical theories. Thereby, it is still a difficulty to form a new prescription and still under the guidance of TCM theories. 3.3

High-throughput screening with multitargets

It is well known that the pharmadynamical function of TCM herbs prescriptions acts in the way of the integral regulation of multicomponents and multitargets. This integral regulation may be the accumulation of multitargets biological effects, the organic chain reaction of multilinks and multitargets, the direct effect of effective medical substance toward the target, or the change of biological characteristics of the target influenced by the internal chemical environment adjustment due to effective medical substances [15]. On the basis of ligands-targets theory and combined with computer technology, high-throughput screening with multitargets was proposed and applied the analysis of the targets or the screen of effective compounds [10]. This method reveals the effective material base and mechanism of TCM herbs prescriptions from cellular, molecular, enzyme and gene level. Biochips are widely applied in the study on material base of TCM herbs prescriptions. Main gene groups of different compound prescriptions are identified in the study in order to analyze the related genes, improve targets, and finally

develop new compounds [16, 17]. The effective material bases of Chinese herbal medicine and herbs prescriptions can be further explored according to the symptom improvement after medicine administration and the histological and biological adjustment with the gene function. The application of high-throughput screening with multitargets should be on the basis of identifying the target and its relation with the therapeutic effects [18]. TCM herbs prescriptions are used to treat syndromes in TCM. At present, the meaning of TCM syndrome has not been entirely revealed. Thus, the target which exactly fits TCM theories is still unable to be identified. However, this technology will contribute to the development of modernization of TCM in the future, the study on classic prescriptions, and also the interpretation of syndrome in TCM. 3.4

Molecular biochromatography

According to modern molecular biological theories, it is believed that Chinese medicine adjusts life-related activities through interactions of neurotransmitters, enzymes, recaptors with chemical constituents, in order to maintain the balance between internal environment and external environment. Therefore, molecular biochromatography is introduced with pharmaceutical chemistry. This method is used to separate and determine active components based on large biomolecule with special identifications. It contains molecular biochromatography with serum albumin stationary phase, biomembrane chromatography, microdialysis sampling coupled with HPLC, and so on. [9] Zou [19] applied molecular biochromatography with human serum albumin and Į-acid glycoprotein as stationary phases to separate active compounds from several kinds of common Chinese herbs. Results showed that effective components, such as ferulic acid and ligustilide of Angelica sinensis (Oliv.) Diels, and capillarisin and scoparone of Herba Artemisia scoparia, remain obviously in the molecular biochromatography. The molecular biochromatography is not only applied in the study on material base of TCM herbs prescriptions but also assisted in the explanation of ADME/Toxic of TCM [20]. Besides, the crude extract can be directly used in this method, which is simple, quick, and repeatable. Thus, this method may be widely used in exploring the material base of TCM herbs prescriptions. 3.5

TCM chromatographic pharmacodynamics

The study of chromatographic pharmacodynamics is on the basis of fingerprint. The research process is composed of the following three steps: (1) A system of TCM mathematic expression of state function relationship, which corresponds to the “syndromes” of animal (or human being), model should be established; (2) A fingerprint of quality or efficiency should be established according to the relationship of indented joint between microcosmic substances and

Zhao Xiaoli et al. / Mode Tradit Chin Med Mater Med, 2009, 11(4): 488–492

effectors corresponding to “syndromes” in TCM, with the target separators of effectors based on affinity chromatograph as its stationary phase and by the application of LC/ MS for the detection of its quality and efficiency; (3) Fundamentals of the effective components of TCM compounds are studied for the relationship of the change between the fingerprint and the state function value of organisms in accordance with the changing tendency of the efficiency value before and after the functioning of effectors (when the concentration of drugs in vivo is zero) [21]. Ning [22] launched a study on the material base of TCM Wuzhuyu decoction with the assistance of chromatographic pharmacodynamics. The analysis revealed that the chemical constituents of peak No. 4, No. 9 (which is identified as Rutaecarpine), No. 10, and No. 12 are in direct proportion with the given pharmacological indexes. It showed that these constituents are the main effective material bases of this prescription. This method bases on fingerprint and takes effectors and efficacy as its main study objects. The breakthrough is considered as the system of TCM mathematic expression of state function and its correspondent “syndromes” in TCM. However, the detailed research idea has not been formed. So this method is still in the starting phase. Its application in the study of material base of TCM herbs prescriptions should be further explored. 3.6

Metabonomics

The effect of Chinese herbs is achieved though the systematic intervention with the human metabolic network with multiefficacy in Chinese herbs. It is featured as multitargets intervention on the systemic level. Thus, in combination with biology, metabonomic with its holism concept was used in the study of compound preparations. The effective material bases of TCM herbs prescriptions are identified through the analysis of all metabolic group changes in the spectrum. The so-called “metabolome” is composed of small molecules metabolites or intermediate products in the progress of gene expression and metabolism [23]. This method is not only applied in the study of the metabolic change of herbal medicine itself but also used in the study of endogenous metabolites induced by herbal medicine, which is helpful in directly understanding the physiological and biochemical situations of the body [24]. Therefore, this method is in accordance with the principle of TCM treatment, which is to adjust the balance between yin and yang of the body. Huang [25] studied the biochemical mechanism of Uncaria-stem Misture for hyperactivity syndrome with metabonomic method. The dynamic changes of endoneurotransmitters (e.g. Ach, DA, NE, and 5-TH) in the serum were determined by HPLC after the oral administration of medicine. Biomarkers were revealed from the type and quantity change of metabonomics according to time. Biochemical substances produced by the holistic effect of medicine is regarded as the

material base of this prescription.

4 New thoughts in study on material base of TCM herbs prescriptions Indications of TCM herbs prescriptions are usually related to many types of diseases. And the therapeutic effects are also related to several pharmacological actions. It is still difficult to reveal the material base of TCM herbs prescriptions systematically and thoroughly. We believe that the main bioactive compounds, which can be identified by serum pharmacology, fingerprint, and pharmocodynamic study, are consisted in the chemical constituents absorbed into blood, tissue, and target site after the interaction of drugs and organisms. Hence, we seek to establish a thought on the study of the main material base, which aims to limit indications on the basis of combination disease with syndrome in the process of the research and development of Tongshaimai pellets, which is subject to National “863” Program. Details of the study are as follows: (1) According to the TCM theories and the research on its efficacy by modern medicine, animal molds and main pharmacological indexes are established, which correspond to the special disease of TCM herbs pre- scriptions; (2) Because of the interaction of drugs and organisms, including biotransformation in gastrointestinal tract and metabolism in tissue or organ, various biological samples were collected by different methods and its fingerprint was established. Then the relationship of biological fingerprint and chemical fingerprint was further analyzed in order to identify the main bioactive compounds. Details of the method are as follows: firstly, the absorbed fingerprint was established through pass perfusion in vivo and molecular biochromatography and then was analyzed for the change of the quantity and the type of chemical constituents on the TCM herbs prescriptions, which are absorbed after and before, to screen the potential active compounds; secondly, the serum fingerprint, which shows the change of the quantity and kinds of chemical constituents absorbed into blood, is established by means of serum pharmacology and serum pharmacochemistry. In addition, the adscription of plasma chemical constituents is identified; thirdly, due to the close correlation with the effect of drug and target areas, the fingerprints of the body fluid of target areas and tissues, which is picked up according to the modern medical theories, are analyzed. After collecting the information of different levels, the interaction of different fingerprints, which is chemical fingerprints and the fingerprints of absorption, serum, and target areas, is explored by some statistical methods, such as bivariate, stepwise, and enter, in order to seek the main material base of TCM herbs prescriptions.

5

Conclusions

With the guidance of TCM theories and clinical effect, the material base of TCM herbs prescriptions is studied by some models of pharmacology, which correspond to the main

Zhao Xiaoli et al. / Mode Tradit Chin Med Mater Med, 2009, 11(4): 488–492

efficacy of TCM, and assisted by the action mechanism of TCM by genomics and proteomics. But in terms of the situation, the study on material base of TCM herbs prescriptions is only that of certain disease, whereas the “syndromes” of TCM is the synthesis function of the intrinsic quality of many diseases. So it is believed that the current study is only a small part of the material base of TCM herbs prescriptions. Recently, chemonics-integrated global systems biology is developed for the study of interaction between TCM and living system on the basis of fundamental principle of TCM and

systems biology, which is able to combine syndromes, prescription, and effect. Therefore, it will be helpful to reveal the material base and action mechanism of TCM herbs prescripttions [26]. The study on the material base of TCM herbs prescriptions shows the significance of studying the TCM fundamentals in the development of revealing compound preparations, improving preparation process, and ensuring clinical efficacy of Chinese medicine.

References [1]

Luo GA, Wang YM. Chemical studies of the compound prescriptions of Traditional Chinese Medicine. World Sci. Technol. Mod. Chin. Mater. Med 1999; 1:16-19.

[2]

[3]

[4]

[5]

Sci. Technol.: Mod. Chin. Mater. Med 2001; 3(5):37-40.

study on material base of medicinal effectiveness of com-

[14] Luo GA, Liang QL, Zhang RL et al. Study of chemical

pound prescriptions of Chinese medicine. World Sci. Technol.:

matteromics and prescription of traditional Chinese medicine.

Mod. Chin. Mater. Med. 2003; 5(6):13-15.

World Sci. Technol.: Mod. Chin. Mater. Med 2006; 8(1):6-15.

Liu JY, Ren JG. Investigation of substantial basis of effects of

[15] Li PT, Qiao YJ, Wang YY et al. Several viewpoints about the

Chinese medicinal compounds. Research and Information on

research methods based on the characteristics of the tradi-

Traditional Chinese Medicine 2004; 6(12): 8-11.

tional Chinese prescription. Journal of Beijing University of

Ma ZC, Gao Y, Tan HL et al. Study of blood-tonifying me-

Traditional Chinese Medicine 1999; 22(5):15-18.

chanisms of Siwutang of traditional Chinese medicine by

[16] Liu WT, Li LH, Dai J. Thinking way and methods of the

technology of TCM moleculomics. World Sci. Technol.: Mod.

biochip technology and Chinese herbs action mechanisms.

Chin. Mater. Med 2005; 7(3):24-28.

Chin. Journal of Basic medicine in Traditional Chinese

Tang WF, Wan MH, Huang X. Tissue pharmacology of

medicine 2008;14(11):845-847.

[8]

[9]

[17] Wang Z, Liu JY, Zhang BL et al. Application of biochip,

36(1): 1-3.

bioinformatics and high-throughput techniques to screen of

Wang XJ. Progress and future developing of the serum phar-

activity of compound Chinese medicines. Chin. J. Chin.

macochemistry of Traditional Chinese Medicine. Chin. J.

Mater. Med 2003; 28(7):686-688.

Chin. Mater. Med 2006; 31(10): 987-297. [7]

developing of the material base of TCM compounds. World

Wu KY, Liang GY, Jin FY et al. Thoughts and methods in

recipe—A new hypothesis. Chin. Tradit. Herb Drug 2005; [6]

Material Medica 1998, 14(3):42-44. [13] Zhang ZY, Zhen XK, Bi YF et al. Progress and future

[18] Wang MQ, Wu B, Yuan SH et al. Investigation of molecular

Jin H, Wang Y, Yan C. Adscription of plasma effective con-

mechanism of TCM compounds. Modern Journal of Inte-

stituents of rat after oral administration of Gegen Qinlian

grated Traditional Chinese and Western Medicine 2003; 12

decoction. Chin. J. Chin. Mater. Med 2008; 33(22): 2687-2691.

(5):449-450.

Sun H, Zhang N, Li LJ et al. Promoting effect of constituents

[19] Kong L, Zou HF, Wang HL et al. Analysis of biologically

in plasma after oral administration of Liuwei Dihuangwan on

active compounds in Chinese medicines by molecular bio-

proliferation of rat osteoblast. Chin. J. Chin. Mater. Med

chromatography with human serum albumin (HSA) sta-

2008; 33(17):2161-2163.

tionary Phase. Chemical Journal of Chinese Universities

Wang XJ, Zhang N, Sun H et al. Protective effect of Liuwei

2000; 21(1):36-40.

Dihuang Wan and its constituents absorbed into blood after oral

[20] Zhang ZQ, Zhang YN, Tian ZJ. Screening models and

administration on hydrocortisone inducing renal deficiency in rat.

techniques for medicine and its application in the studies on

Chin. Exp. Tradit. Med. Form 2008, 14(2): 33-37.

the active components of Chinese medicine. Chin. J. Chin.

[10] Guo LM, Wang CY, Gu QQ et al. Progression on studies of the effective components of decoction. Chin. Tradit. Pat. Med 2007; 29(1):118-121. [11] Cao ZQ. New thinking about study of pharmacodynamic material basis and functional mechanism in Chinese material medica. Journal of Shanghai University of Traditional Chinese Medicine 2000; 14(1):63-93. [12] Yang K, Guo L, Zhou MM. Discussion on method for studying combinatorial chemical research of traditional Chinese prescriptions. Pharmacology and Clinics of Chinese

Mater. Med 2003; 28 (10):907-910. [21] He FY, Luo JF, Liu WL. Preliminary exploration of orlentation and method of chromatographic pharmacodynamics of traditional Chinese medicine. World Sci. Technol.: Mod. Chin. Mater. Med 2004; 6(6):44-50. [22] Ning LL, Bi KS, Wang R et al. Methodological study on the material basis for the efficacy of the traditional Chinese medicine Wu ZHU YU decoction. Acta Pharm. Sin 2000; 35 (2):131-134. [23] Liu CX. Systems biology and modern research of traditional

Zhao Xiaoli et al. / Mode Tradit Chin Med Mater Med, 2009, 11(4): 488–492

Chinese medicines. Journal of Tianjin University of Traditional Chinese Medicine 2006; 25 (4):191-194.

hyperactivity syndrome by metabonomic method. Chin. Tradit. Herb Drug 2005; 36 (3):398-402.

[24] Liu CX. Significance of metabonomics in modern research

[26] Luo GA, Liang QL, Liu QF et al. Chemomics-integrated

of Chinese materia medica. Chin. Tradit. Herb Drug 2004;

global systems biology: A holistic methodology of study on

35 (6):10.6-50.6.

compatibility and mechanism of formulas in traditional

[25] Huang YR, Wei GL, Long H et al. Pharmacodynamics and its biochemical mechanism of Uncaria-stem Mistura for

Chinese medicine. World Sci. Technol.: Mod. Chin. Mater. Med 2007; 9(1):10-15.