An in vivo analysis of the therapeutic and synergistic properties of Chinese medicinal formula Yin-Chen-Hao-Tang based on its active constituents
Graphical abstract
Traditional Chinese medicine (TCM) formula usually takes multi-component to produce a sufficient effect at low doses, exert a more robust therapeutic effect than any individual compounds by hitting multiple targets.
Introduction
During the past few years, the pharmaceutical industry has seen a shift from the “1 disease, 1 target, 1 drug” and “1 drug fits all” approaches to the pursuit of combination therapies that include more than one active ingredient [1], [2], [3]. The complexity of medicine suggests that treatment protocols should be carefully designed, and the construction of a prescription is an art in fighting disease. Increasing evidence demonstrates that, in treating illnesses, including human acute promyelocytic leukemia [4], [5], cancer [6], HIV [7] and chronic hepatitis C virus [8], treatment regimens containing multiple drugs with distinct but related mechanisms can usually amplify the therapeutic efficacies of each agent, leading to maximal therapeutic efficacy with minimal adverse effects, represent a triumph for modern medicine [9], [10]. Interestingly, TCM is a unique medical system assisting the ancient Chinese in dealing with disease, and has advocated combinatory therapeutic strategies for 2500 years by prescriptions called formulae [11]. Typically, formulae consist of several types of medicinal herbs or minerals, in which one represents the principal component, and others serve as adjuvant ones to assist or facilitate the effects of the principal component [12], [13]. The therapeutic efficacy of TCM usually attributed to the synergistic property, improving the therapeutic efficacy, among multiple herbs and constituents which was named as “formula compatibility”. It is believed that multiple components in the formulae could hit multiple targets and exert synergistic therapeutic efficacies [12], [14]. A growing body of evidence has demonstrated that multidrug combinations are increasingly important in modern medicine [15], [16]. However, the precise mechanisms through which formulae function are poorly understood and must be addressed using a modern approach and will certainly further promote the reasonable and effective application of TCM.
Yin-Chen-Hao-Tang (YCHT), which was recorded in “Shanghanlun” and is a classic resource on TCM written by Zhongjing Zhang (150–215 A.D.), is one of the most famous Chinese herbal formulae. YCHT consists of Artemisia annua L. (the monarch herb; D), Gardenia jasminoides Ellis (the minister herb; G), and Rheum Palmatum L. (the assistant and servant herb; R) and has been used for more than a thousand years to treat jaundice and liver disorders [17]. Pharmacological studies and clinical practices have shown that it can be used clinically to treat cholestasis, hepatitis C, primary biliary cirrhosis, liver fibrosis, and cholestatic liver diseases [18], [19], [20], [21]. Interestingly, D has been shown to be effective in treating HI by exerting hepato-protectivity and contributes directly to the therapeutic effect of YCHT [22]. G, a major component of fruits of Gardenia jasminoids Ellis, has various pharmacological activities, including anti-oxidant, anti-inflammatory and hepatoprotective effects [23], [24], [25]. A recent study showed that rhein, a metabolite of anthranoidsand major components of Rheum officinale Baillon, contributes to restrict the liver fibrosis [26], [27], [28]. D, G and R were adopted as marker compounds for quality control in the manufacturing process of this medicine in our facility [29]. Of note, a recent report have reported that with formula change, every herbal drug exhibited different effects on the pharmacokinetics of D and G in normal rats, all of which could contribute to the special priority of compatible administration of YCHT [30].
Generally, the curative effect of TCM is an integrative result of a number of bioactive compounds. Understanding synergistic effects of DGR represents an even greater challenge. Up to now, results from in vivo pharmacokinetic studies performed on D, G and R (chemical structures of three markers were shown in Fig. 1.) in some prescriptions, have been carried out [31], [32], [33], [34], [35]. However, synergistic property on the combined pharmacokinetic and biochemical strategy has not been explored yet. Previous studies (medicinal substances level) showed that with formula change, every herbal drug exhibited different effects on the pharmacokinetics of D and G in normal rats, all of which could contribute to the special priority of compatible administration of YCHT [30]. In trying to approach the rationale of formula design in TCM, here we use the treatment of HI with DGR as a working model. D, G, and R were used as active compounds of Artemisia capillaries Thunb, Gardenia jasminoids Ellis, and Rheum officinale Baillon, respectively, and the therapeutic and synergistic effects of DGR combination on HI were elucidated.
Section snippets
Materials and reagents
Acetonitrile and methanol, HPLC grade, were purchased from Dikma Technology Corporation (Richmond Hill, Ontario, Canada). Deionized water was purified on a Milli-Q system (Millipore, Bedford, USA). Formic acid and phosphoric acid, analytical grade, were obtained from Beijing Reagent Company (Beijing, China). Carbon tetrachloride (A.R, Batch No. 200904081) was purchased from Chemicals Factory (Tianjin, PR China). Glycerol was supplied from Chemicals Factory (Shanghai, PR China). Olive oil
Method validation of pharmacokinetics
To develop a simple and popular method for D, G, R assay in rat plasma for pharmacokinetic studies, HPLC with UV detection was selected as the method of choice. The chromatograms for blank rat plasma, D, G and R in rat plasma after oral administration, and D, G and R in methanol are shown in Fig. 2. These chromatographic conditions revealed no biological substances that would significantly interfere with the accurate determination of D G and R. The retention time for D, G and R is approximately
Discussion
Currently, rather than focusing on a single target, a paradigm shift is occurring that is sparking new interest in agents that modulate simultaneously multiple targets with fewer adverse effects and lower toxicity. Combination medicines with multi-effect pathways and multi-effect targets tend to be more effective than single drugs and may help to address many treatment-related challenges [36]. However, continued progress will be essential if we are to develop effective, customized multi-drug
Acknowledgments
This work was supported by grants from the Key Program of the Natural Science Foundation of the State (Grant No. 90709019), the National Key Program on the Subject of Drug Innovation (Grant No. 2009ZX09502-005), and the National Program for Key Basic Research Projects in China (Grant No. 2005CB523406).
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2020, Journal of Pharmaceutical and Biomedical AnalysisCitation Excerpt :Traditionally, TCMPs are composed of multiple herbs that follow the principle of compatibility efficacy guided by traditional Chinese medicine (TCM) theory, and display a synergistic action so as to obtain the curative effect [7,8]. The synergistic action could achieve efficacy enhancing and toxicity reducing by a series of complicated interactions in the existence of thousands of constituents, such as solubilization, precipitation, etc. [7–9]. Accordingly, TCMP, which is composed of various herbal medicines with complicated chemical composition, should be regarded as an organic whole to take chemical contribution of individual herbal medicine and structural diversity into consideration as more as possible, so as to comprehensively characterize its chemical profile.