| Quality control and evaluation is one of the key issues for the modernization of Chinese materia medicas (CMMs), the efficient constituents identification is the foundation of CMMs'Quality control. However, current pattern of quality control for CMMs mainly focused on the qualitative and quantitative determination of chemical components, could assure neither the quality stability of CMMs, nor the safety and efficacy.To seek a new breakthrough of the efficient constituents identify and quality control for CMMs, its current status and problems with qulity control of CMMs, significance and practical importance of screening/ definite pharmacodynamic constituents of CMMs for the quality control were all studied; And a strategy combining spectrum-effect relationship with quantity-effect relationship for screening and analysis of bioactive components and quality control of CMMs named "knock out & knock in" was introduced and demonstration, which could supplement and consummate the current system of the quality control and evaluation for CMMs. Besides the pattern for the bioactive components screening and quality control of the "Gene Diagmosis& Treatment" was introduced to refer. In this article, Curcuma longa L. was studied as a mould due to its universal usage in clinic and relative clarity of chemical components to establish a pattern and method of the quality control for CMMs which was based on the strategy of "knock out & knock in". This study was provided a theory and reference for quality control of CMMs for development and standards. The main results and conclusion are summarized below:(1) In the view of spectrum-effect relationship, the "pharmacodynamic constituents groups" which are related with traditional/clinical pharmacodynamic action were analyzed and investigated. Ultra-performance liquid chromatography (UPLC) was used for analysis of chemical constituents of the 13 batches Curcuma sample, and the chemical fingerprints were established. Antioxidant phannaco-activity in vivo of curcuma were evaluation by the tradition pharmacological method, the "spectra-effect" relationships were established combined with chemical fingerprints, and preliminarily screening antioxidant composition/portion. Canonical correlation analysis results indicated Curcuminoids were active composition/portion of antioxidant activity from total extracts of Curcuma.(2) Screening and identification of the antioxidant activity constituents of Curcuma longa L. based on the strategy of target constituents "knock-out & knock-in", and a method for evaluation antioxidant activity in vitro were preliminarily established. The antioxidant activity of samples that were obtained by "knock-out" from the alcohol extract of Curcuma longa L., was determined, key pharmacodynamic constituents (KPCs) were identified and the regulation of multi-components were elucidated. The free radical scavenging rate which is related with the antioxidant activity of C longa was used as evaluation index when the total extract of C.longa was taken as a reference. The results showed that Cur, DMC and BDMC were contribution extent were 61.92%,11.09% and 9.10% of the total extract respectively, showing the composition and content of active substances of scavenging DPPH free radical.the results were consistent with the investigation had validated, which is suggested that the target constituent "knock-out" strategy was applied to the KPCs screen were reasonable, and have good feasibility.(3) Then, the target constituent "knock-in" strategy was introduced to study the quantity-activity relationships of the KPCs in C.longa. The contents range was delimited for quality control, which could provid some reference for controlling the quality of C.longa and the new quality criteria related with clinical curative effect were established. The mutual relationships of multi-components were elucidated based on "Knock-out". Then, the quantity of the target constituent was added into the minus extract, respectively. And the quantity-activity relationship was established and the range might be delimited for the aim to quality control. The results suggested that the contents of Curcumin should not be lower than 0.96%, and demethoxycurcumin should not be lower than 0.30%.This study could provid some reference for new quality criteria of C.longa.(4) Bases on traditional effectiveness of Curcuma longa L., the anticoagulation and antithrombotic activity were applied to investigate the correlation relationship between each constituent and anticoagulation activity by the target constituent "knock-out" strategy, and KPCs were identified, the results were consistent with the investigation had reported, this results suggested that the target constituent "knock-out" strategy was applied to the KPCs screen were reasonable, and have good feasibility.(5) In this study, the pattern and strategy for KPCs identify and quality control of CMMs based on target constituents "knock-out & knock-in" was produced and established, the results indicated that this strategy was applied to the KPCs screen and quality contol were scientific, reasonable, and have good feasibility. According to above results, the quality criteria protocol of C.longa based on pharmacodynamic action was preliminarily stipulated which provide some idea and technical support for effectiveness-related quality control for C.longa.In summary, This strategy of target constituent "Knock out & Knock in" is not only identifying pharmacodynamic constituents objectively and validity; Also it is well coincident with the characteristic of holistic and systemic of CMMs. This strategy can more precisely and quickly, and provide some idea and technical support for recognize the KPCs and effectiveness-related quality control for C.longa and other CMMs. |
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