Traditional Chinese medicine efficacy substance screening method based on 6-dimensional cobweb model

Abstract

The invention discloses a traditional Chinese medicine efficacy substance screening method based on a 6-dimensional cobweb model, and belongs to the technical field of medicine. Based on a UHPLC (Ultra High Performance Liquid Chromatography) analysis result, tanshinol, chlorogenic acid, procyanidine B2, epicatechin, hyperoside, isoquercitrin, rosmarinic acid, alkannic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid and ursolic acid are used as specific candidate functional components of the salvia miltiorrhiza-hawthorn medicine pair; and constructing a cobweb model based on data of the candidate functional components in six dimensions, namely an effectiveness dimension, a compatibility environment dimension, a transmission and traceability dimension, a content testability dimension, a network pharmacology value dimension and a molecular docking target binding activity dimension, so as to obtain the key medicinal effect substances of the salvia miltiorrhiza-hawthorn drug pair. The invention provides theoretical support for revealing the compatibility theory of the salvia miltiorrhiza-hawthorn medicine pair, formulating the quality evaluation standard and clinical application of the salvia miltiorrhiza-hawthorn medicine pair.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine and relates to a method for screening active ingredients of traditional Chinese medicine based on a six-dimensional spider-web model. Background Art

[0002] The Danshen-Hawthorn herb pair is a classic prescription from "Shi Jinmo's Paired Herbs", consisting of two traditional Chinese medicines, Danshen and Hawthorn. It is a classic clinical combination for treating ischemic heart disease based on the traditional Chinese medicine theory of promoting blood circulation to remove blood stasis. Among them, Danshen is slightly cold in nature and bitter in taste, and belongs to the heart and liver meridians, with the effects of promoting blood circulation to remove blood stasis, cooling blood and dissipating carbuncles, and clearing the heart and relieving vexation; Hawthorn is slightly warm in nature and sour and sweet in taste, and belongs to the spleen, stomach and liver meridians, with the effects of promoting digestion and resolving food stagnation, promoting qi circulation and dispersing stasis, and reducing turbidity and lipid-lowering. The combination of the two can produce a synergistic effect, significantly improving the symptoms of chest pain due to blood stasis in the heart, and has important application value in the clinical treatment of cardiovascular diseases.

[0003] The active ingredients of traditional Chinese medicine refer to the chemical components or component groups in traditional Chinese medicine and its compound prescriptions that can characterize clinical efficacy. The research content includes key scientific issues such as the identification of chemical components or component groups, the analysis of in vivo metabolism processes, and the confirmation of action targets. The traditional research mode mainly uses the separation and purification of single components combined with activity screening, which is difficult to comprehensively explain the overall action characteristics of traditional Chinese medicine of "multiple components - multiple targets - multiple pathways". Therefore, establishing a research method that conforms to the holistic characteristics of traditional Chinese medicine and systematically revealing the synergistic action mechanism of each component in traditional Chinese medicine compound prescriptions has great theoretical significance and application value for promoting the modern research of traditional Chinese medicine.

[0004] The spider-web model is a multivariate statistical analysis method based on the principle of radar chart, which can realize the comprehensive evaluation of complex systems through the visualization of multi-dimensional evaluation indexes. At present, the research on the active ingredient basis of the Danshen-Hawthorn herb pair against myocardial ischemia is not sufficient, and its scientific connotation of compatibility and quality control standards need to be clarified urgently. Therefore, exploring and developing a method for screening active ingredients of traditional Chinese medicine based on the spider-web model will provide important technical support for clarifying the compatibility rules of the Danshen-Hawthorn herb pair, establishing a scientific quality evaluation system, and guiding rational clinical medication. Summary of the Invention

[0005] To solve the above technical problems, the present invention provides a method for screening the effective substances of traditional Chinese medicine based on a 6-dimensional cobweb model. Based on the UHPLC analysis results, taking danshensu, chlorogenic acid, procyanidin B2, epicatechin, hyperoside, isoquercitrin, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid, and ursolic acid as candidate effective components, a "cobweb" model is constructed based on the six dimensions of the candidate effective components in the dimensions of "effectiveness", "compatibility environment", "transmission and traceability", "content measurability", "degree value" dimension of network pharmacology, and "target binding activity" dimension of molecular docking to obtain the effective substances of the danshen-hawthorn herb pair. The present invention provides theoretical support for revealing the compatibility theory of the danshen-hawthorn herb pair, formulating the quality evaluation standard of the danshen-hawthorn herb pair, and the clinical application of the danshen-hawthorn herb pair.

[0006] To achieve the technical purpose of the present invention, on the one hand, the present invention provides a method for screening the effective substances of traditional Chinese medicine based on a 6-dimensional cobweb model, wherein the traditional Chinese medicine in the effective substances of traditional Chinese medicine is the danshen-hawthorn herb pair, and the method for screening the effective substances of traditional Chinese medicine includes: Composing a 6-dimensional characteristic network based on the concept of the "five principles" in traditional Chinese medicine quality markers; normalizing the data of each dimension in the 6-dimensional characteristic network to obtain quantitative values; respectively linking the quantitative values of the same candidate effective component in different dimensions to construct a regression region of the candidate effective component; sorting the areas of the regression regions, and screening candidate effective components with a regression region area higher than 0.30 to obtain the effective substances of traditional Chinese medicine; Specifically, the candidate effective components are composed of danshensu, chlorogenic acid, procyanidin B2, epicatechin, hyperoside, isoquercitrin, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid, and ursolic acid; Specifically, the 6-dimensional characteristic network is composed of the dimensions of "effectiveness", "compatibility environment", "transmission and traceability", "content measurability", "degree value" dimension of network pharmacology, and "target binding activity" dimension of molecular docking of the candidate effective components.

[0007] Furthermore, in the method for screening the effective substances of traditional Chinese medicine based on a 6-dimensional cobweb model provided by the present invention, the method for obtaining the data of the "effectiveness" dimension includes: Based on orthogonal partial least squares, associating the levels of myocardial injury markers with the peak areas of the common peaks in the danshen-hawthorn herb pair to obtain the variable projection importance values of the compounds corresponding to the common peaks and the anti-myocardial injury activity; Based on grey relational analysis, the levels of myocardial injury markers were correlated with the peak areas of the common peaks in the Salvia miltiorrhiza - Crataegus pinnatifida pair, and the correlation degree values between the compounds corresponding to the common peaks and the anti - myocardial injury activity were obtained; The variable importance in projection values and correlation degree values of the compounds corresponding to the common peaks and the anti - myocardial injury activity were normalized to obtain the data of the "effectiveness" dimension.

[0008] Specifically, the myocardial injury markers are lactate dehydrogenase and creatine kinase isoenzyme.

[0009] Furthermore, in the method for screening the effective substances of traditional Chinese medicine based on the 6 - dimensional cobweb model provided by the present invention, the method for obtaining the data of the "compatibility environment" dimension includes: The weight of the candidate efficacy components that only exist in Salvia miltiorrhiza in the Salvia miltiorrhiza - Crataegus pinnatifida pair was set to 25, the weight of the candidate efficacy components that only exist in Crataegus pinnatifida was set to 75, and the weight of the candidate efficacy components that exist in both Salvia miltiorrhiza and Crataegus pinnatifida was set to 100; The weights of the candidate efficacy components were normalized to obtain the data of the "compatibility environment" dimension.

[0010] Furthermore, in the method for screening the effective substances of traditional Chinese medicine based on the 6 - dimensional cobweb model provided by the present invention, the method for obtaining the data of the "transmission and traceability" dimension includes: Analyze the distribution of the candidate efficacy components in in vitro samples, serum samples and heart tissue samples; Set the weight of the distribution of the candidate efficacy components in in vitro samples to 20, the weight of the distribution of the candidate efficacy components in serum samples to 80, and the weight of the distribution of the candidate efficacy components in heart tissue samples to 100. After normalization, the data of the "transmission and traceability" dimension are obtained.

[0011] Specifically, the present invention uses chromatography - mass spectrometry coupling technology to analyze and identify the in vitro chemical components of the Salvia miltiorrhiza - Crataegus pinnatifida pair extract, the serum - migrating components and the heart - tissue - distributing components after oral administration. A total of 10 prototype components are found to exist in both serum and heart tissue, namely tanshinone ⅡA, lithospermic acid and its isomers, rosmarinic acid, salvianolic acid A, quercetin, dimethyl lithospermate, lithospermic acid, salvianolic acid B, dihydrotanshinone Ⅰ and cryptotanshinone. The above components can be rapidly distributed to the heart tissue through blood circulation to play a myocardial protection role. Components that only exist in serum are also obtained, including danshensu, chlorogenic acid, hyperoside and isoquercitrin.

[0012] Furthermore, in the method for screening the effective substances of traditional Chinese medicine based on the 6 - dimensional cobweb model provided by the present invention, the method for obtaining the data of the "measurability of content" dimension includes: Danshen and Hawthorn herbs from different purchase sources are compounded according to the mass ratio of Danshen:Hawthorn = 1:3 to obtain the Danshen-Hawthorn herb pair; Determine the content of candidate efficacy components in the Danshen-Hawthorn herb pair; Normalize the content of the candidate efficacy components to obtain the data of the "content measurability" dimension.

[0013] Specifically, in the present invention, the content of candidate efficacy components in 14 batches of Danshen-Hawthorn herb pairs purchased from different manufacturers was determined according to the UHPLC method, and the average values of 16 candidate efficacy components in the extracts of 14 batches of herbs were calculated respectively. This value reflects the content level of each component in the Danshen-Hawthorn herb pair. Finally, data normalization was performed as the data of the content measurability dimension.

[0014] Furthermore, in the method for screening traditional Chinese medicine efficacy substances based on the 6D cobweb model provided by the present invention, the method for obtaining the data of the "degree value" dimension of network pharmacology includes: Visualize the candidate efficacy components and action targets to construct a component-target network; Apply the Analyzer tool to calculate network topology parameters to obtain the data of the "degree value" dimension of network pharmacology.

[0015] Specifically, in the present invention, 386 action targets related to the Danshen-Hawthorn herb pair were predicted through the TCMSP, Swiss Target Prediction, and Batman-Tcm platforms. The candidate efficacy components and action targets were imported into the Cytoscape 3.7.2 software for visualization processing to construct a "component-target" network, and the Analyzer tool was used to calculate network topology parameters to obtain the degree values of the candidate efficacy components.

[0016] Furthermore, in the method for screening traditional Chinese medicine efficacy substances based on the 6D "cobweb" model provided by the present invention, the method for obtaining the data of the "target binding activity" dimension of molecular docking includes: Perform molecular docking of the candidate efficacy components with the core targets, and calculate the binding free energy of the selected efficacy components and the core targets; Normalize the binding free energy to obtain the data of the "target binding activity" dimension of molecular docking; The core targets consist of STAT3, SRC, TP53, JUN, and AKT1.

[0017] On the other hand, the present invention claims the efficacy substances screened by the above method for screening traditional Chinese medicine efficacy substances based on the 6D cobweb model, which are composed of salvianolic acid B, tanshinone IIA, dihydrotanshinone I, rosmarinic acid, isoquercitrin, lithospermic acid, cryptotanshinone, ursolic acid, procyanidin B2, and hyperoside.

[0018] In addition, the present invention claims the application of the above-mentioned pharmacodynamic substances in establishing the quality evaluation standard for the Salvia miltiorrhiza - Crataegus pinnatifida drug pair.

[0019] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages: (1) The present invention provides a screening method for traditional Chinese medicine pharmacodynamic substances based on a 6 - dimensional spider web model. A 6 - dimensional characteristic network is composed based on the concept of the "five principles" in traditional Chinese medicine quality markers, specifically including six dimensions: the "effectiveness" dimension, the "compatibility environment" dimension, the "transmission and traceability" dimension, the "content measurability" dimension, the "degree value" dimension of network pharmacology, and the "target binding activity" dimension of molecular docking. The measurement data of each dimension are subjected to min - max normalization processing to eliminate the analysis deviation caused by big data fluctuations. Finally, a radar chart is drawn using R language, and the area of the regression region is calculated. The candidate efficacy components with a regression region area higher than 0.30 are sorted out, and the candidate efficacy components are the traditional Chinese medicine pharmacodynamic substances.

[0020] (2) Through a screening method for traditional Chinese medicine pharmacodynamic substances based on a 6 - dimensional spider web model, the present invention has screened out that the pharmacodynamic substances in the Salvia miltiorrhiza - Crataegus pinnatifida drug pair are composed of salvianolic acid B, tanshinone ⅡA, dihydrotanshinone Ⅰ, rosmarinic acid, isoquercitrin, lithospermic acid, cryptotanshinone, ursolic acid, procyanidin B2, and hyperoside. The pharmacodynamic substances screened out by the present invention are convenient for formulating the quality evaluation standard for the Salvia miltiorrhiza - Crataegus pinnatifida drug pair, and ensure the efficacy stability and quality controllability of the Salvia miltiorrhiza - Crataegus pinnatifida drug pair in clinical applications. Brief Description of the Drawings

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following - described drawings are only some embodiments of the present invention.

[0022] Figure 1 It is the UHPLC chromatogram of the Salvia miltiorrhiza - Crataegus pinnatifida drug pair at a detection wavelength of 280 nm. Figure 1 In it, A is the UHPLC chromatogram of the mixed reference substances at a detection wavelength of 280 nm; Figure 1 In it, B is the UHPLC chromatogram of the test sample of the Salvia miltiorrhiza - Crataegus pinnatifida drug pair at a detection wavelength of 280 nm. The attached drawing reference numerals are as follows: Peak 1: salvianolic acid B; Peak 2: 5 - hydroxymethylfurfural; Peak 3: danshensu; Peak 4: procyanidin B2; Peak 5: epicatechin; Peak 6: salvianolic acid A; Peak 7: dihydrotanshinone Ⅰ; Peak 8: tanshinone Ⅰ; Peak 9: cryptotanshinone; Peak 10: tanshinone ⅡA.

[0023] Figure 2It is the UHPLC chromatogram of Danshen-Hawthorn medicine at a detection wavelength of 335 nm. Figure 2 In it, A is the UHPLC chromatogram of the mixed reference substances at a detection wavelength of 335 nm; Figure 2 In it, B is the UHPLC chromatogram of the Danshen-Hawthorn medicine pair test sample at a detection wavelength of 335 nm. The attached drawing reference numerals are as follows: Peak 11: Chlorogenic acid; Peak 12: Hyperoside; Peak 13: Isoquercitrin; Peak 14: Rosmarinic acid; Peak 15: Lithospermic acid peak.

[0024] Figure 3 It is the UHPLC chromatogram of Danshen-Hawthorn medicine at a detection wavelength of 210 nm. Figure 3 In it, A is the UHPLC chromatogram of the mixed reference substances at a detection wavelength of 210 nm; Figure 3 In it, B is the UHPLC chromatogram of the Danshen-Hawthorn medicine pair test sample at a detection wavelength of 210 nm. The attached drawing reference numerals are as follows: Peak 16: Oleanolic acid; Peak 17: Ursolic acid.

[0025] Figure 4 It is the screening diagram of the active ingredients of Danshen-Hawthorn medicine pair based on the "spider web" model. Figure 4 In it, A is the histogram of the regression area ranking of the "spider web" model; Figure 4 In it, B is the 6-dimensional feature network result of the "spider web" model. Specific implementation manners

[0026] Next, the technical solutions of the present invention will be described in conjunction with the embodiments. However, the present invention is not limited to the following embodiments. The experimental methods and detection methods described in each embodiment are all conventional methods unless otherwise specified; the reagents and materials described are all commercially available unless otherwise specified.

[0027] Example 1 This example provides the construction of each dimension of the 6-dimensional feature network in the "spider web" model.

[0028] The present invention composes a 6D feature network based on the concept of "five principles" in traditional Chinese medicine quality markers. The candidate efficacy components are determined by the "uniqueness" of the components. The 6D feature network specifically includes: the "efficacy" dimension of the candidate efficacy components, the "compatibility environment" dimension, the "transmission and traceability" dimension, the "measurability" dimension of the content, the "degree value" dimension of network pharmacology, and the "target binding activity" dimension of molecular docking. The "efficacy" dimension is evaluated by integrating the VIP values of compounds obtained by two spectral efficacy analysis methods, OPLS and GRA, and the correlation values of compounds with myocardial injury serum markers LDH and CK-MB; the "compatibility environment" dimension is evaluated by the compatibility ratio of the herb pair; the "transmission and traceability" dimension is evaluated by detecting the distribution of candidate efficacy components in vitro and in vivo (rat plasma, rat heart); the "measurability" dimension of the content is evaluated by measuring the average content of candidate efficacy components in extracts of multiple batches of herb pairs; the "degree value" dimension of network pharmacology is evaluated by constructing a "component-target" network and calculating the network topology parameter degree; the "target binding activity" dimension of molecular docking is evaluated by calculating the binding energy of candidate efficacy components with core targets using molecular docking.

[0029] 1. Screening of candidate efficacy components based on the "uniqueness" of components Previously, UHPLC was used to analyze the chemical components of the extract of the Salvia miltiorrhiza-Fructus Crataegi herb pair (the mass ratio of Salvia miltiorrhiza to Fructus Crataegi is 1:1) at different wavelengths, as Figures 1 to 3 shown. After comparison with reference substances, a total of 17 chemical components were finally identified, including Peak 1: salvianolic acid B, Peak 2: 5-hydroxymethylfurfural, Peak 3: danshensu, Peak 4: procyanidin B2, Peak 5: epicatechin, Peak 6: salvianolic acid A, Peak 7: dihydrotanshinone I, Peak 8: tanshinone I, Peak 9: cryptotanshinone, Peak 10: tanshinone IIA, Peak 11: chlorogenic acid, Peak 12: hyperoside, Peak 13: isoquercitrin, Peak 14: rosmarinic acid, Peak 15: lithospermic acid, Peak 16: oleanolic acid, Peak 17: ursolic acid. Among them, Peak 2 was identified as 5-hydroxymethylfurfural, which is an organic compound usually formed by the dehydration of reducing sugars during heating-related preparation processes. Although 5-hydroxymethylfurfural has been reported to have other activities such as antioxidant, anti-allergic, anti-hyperuricemia, and anti-cancer activities, it is not a specific compound of Salvia miltiorrhiza and Fructus Crataegi, and has been detected in many herbs, honey, and sugary foods. Therefore, the other 16 components were selected as the candidate efficacy components of the Salvia miltiorrhiza-Fructus Crataegi herb pair.

[0030] 2. Evaluation based on the "efficacy" and "compatibility environment" dimensions Previously, SIMCA 14.1 software and SPSSPRO online software were used for spectral-effect relationship analysis. The levels of myocardial injury markers in the serum of mice in the treatment groups with different compatibility ratios of Salvia miltiorrhiza - Hawthorn fruit herb pairs (the mass ratios of Salvia miltiorrhiza to Hawthorn fruit were 1:0, 0:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5) were related to the peak areas of the common peaks in the corresponding samples. The serum myocardial injury markers were lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB). Orthogonal partial least squares (OPLS) model and grey relational analysis (GRA) were used for spectral-effect relationship analysis, and the optimal compatibility ratio of Salvia miltiorrhiza - Hawthorn fruit herb pair was screened out as 1:3. The variable importance in projection (VIP) values obtained from the OPLS model and the correlation values of the compounds with LDH and CK-MB obtained from the GRA model (Table 1) were normalized as the "effectiveness" dimension for evaluating the efficacy of the compounds. For different drugs, with different compatibility ratios, there will be differences in the efficacy and material basis. Starting from the "compatibility environment" of the herb pair, the weights of the candidates of Salvia miltiorrhiza and Hawthorn fruit were set as 25 and 75 respectively, and the common components of the two were set as 100. Finally, the normalization of the weights of each component was carried out.

[0031] Table 1 Spectral-effect analysis table of the anti-myocardial injury activity of Salvia miltiorrhiza - Hawthorn fruit herb pair

[0032] 3. Evaluation based on the "transmission and traceability" dimension The components of the Salvia miltiorrhiza - Hawthorn fruit herb pair need to enter the blood or the heart to play their cardioprotective role. Therefore, an evaluation of the "transmission and traceability" dimension of the candidate compounds entering the blood and the heart was carried out. Previously, comprehensive use of chromatography-mass spectrometry technology was used to analyze and identify the in vitro chemical components of the extract of the Salvia miltiorrhiza - Hawthorn fruit herb pair, the serum migratory components and the heart tissue distribution components after oral administration. A total of 10 prototype components were found to exist in both serum and heart tissue, namely tanshinone ⅡA, lithospermic acid and its isomers, rosmarinic acid, salvianolic acid A, quercetin, dimethyl lithospermate, lithospermic acid, salvianolic acid B, dihydrotanshinone Ⅰ and cryptotanshinone. The above components can be rapidly distributed to the heart tissue through blood circulation to play a myocardial protection role. Components only present in serum were also obtained, including danshensu, chlorogenic acid, hyperoside, and isoquercitrin. The weights of the candidates that can enter the heart and blood were set as 100 and 80 respectively, and the weights of the candidates that can only be detected in in vitro samples were set as 20. Finally, the normalization of the weights of each component was carried out.

[0033] 4. Evaluation based on the "measurability" dimension of content The measurability of the content of traditional Chinese medicine components is an important factor in ensuring the quality and evaluation of medicinal materials. Therefore, the average content is selected as the evaluation index from the perspective of measurability. In the early stage, through methodological verification, the contents of candidate efficacy components in 14 batches of Salvia miltiorrhiza - Hawthorn fruit herb pairs (pairing the Salvia miltiorrhiza and Hawthorn fruit herbs from the same manufacturer) purchased from different manufacturers were determined according to the UHPLC method. The average values of 16 candidate efficacy components in the extracts of 14 batches of medicinal materials were calculated respectively (Table 2), and this value reflects the content level of each component in the Salvia miltiorrhiza - Hawthorn fruit herb pair. Finally, data normalization was carried out to serve as the "content dimension".

[0034] Table 2 Contents of candidate efficacy components in 14 batches of Salvia miltiorrhiza - Hawthorn fruit herb pairs

[0035] 5. Evaluation of the "component - target" network "degree value" dimension In the early stage, 386 related action targets of the Salvia miltiorrhiza - Hawthorn fruit herb pair were predicted through the TCMSP, Swiss Target Prediction, and Batman - Tcm platforms. The candidate efficacy components and action targets were imported into the Cytoscape 3.7.2 software for visualization processing to construct a "component - target" network, and the Analyzer tool was used to calculate the network topology parameters to obtain the degree values of the candidate efficacy components. The larger the degree value, the higher the importance of the component. The obtained degree values of the candidate efficacy components are 54 for ursolic acid, 51 for salvianolic acid B, 41 for tanshinone IIA, 39 for tanshinone I, 34 for rosmarinic acid, 32 for dihydrotanshinone I, 31 for cryptotanshinone, 26 for lithospermic acid, 18 for oleanolic acid, 12 for salvianolic acid A, 11 for danshensu, 10 for hyperoside, 7 for procyanidin B2, 7 for epicatechin, 4 for isoquercitrin, and 2 for chlorogenic acid. Finally, the degree values of the candidate efficacy components were normalized to serve as the "degree value" dimension.

[0036] 6. Evaluation of the "target binding activity" dimension The candidate bioactive components were subjected to molecular docking with the top five core disease targets screened by network pharmacology (STAT3, SRC, TP53, JUN, and AKT1, among which STAT3 is one of the key targets involved in myocardial ischemia injury) to evaluate the specific binding activity dimension between the candidate bioactive components and the disease targets. The two-dimensional structural formulas of the candidate bioactive components were obtained from the PubChem database, and the three-dimensional molecular models were constructed using Chem3D software and exported as mol2 format files. Subsequently, the crystal structures of the core targets were downloaded from the PDB database, and the structures were preprocessed using PyMOL software (removing non-critical components such as water molecules and phosphate groups). AutoDockTools 1.5.6 was used for preprocessing the molecular structures, including format conversion (pdb→pdbqt) and active site identification. Rigid docking calculations of the binding free energy were performed using Autodock Vina, and flexible docking was performed using Discovery Studio 2019 to obtain the LibDockScore to comprehensively evaluate the ligand-receptor interactions. According to the calculation results of Autodock Vina (Table 3), complexes with a binding energy ≤ -5.0 kcal / mol were considered stable binding systems. The three-dimensional conformations were visualized using PyMOL software, and the key binding sites were analyzed in combination with two-dimensional interaction diagrams. Finally, by normalizing the comprehensive binding energies of the candidate bioactive components with the 5 targets, the data for the "target binding activity" dimension were obtained.

[0037] Table 3 Binding energies of candidate bioactive components and core disease targets

[0038] Example 2 This example provides the establishment of the "spider web" model.

[0039] The six dimensions of the OPLS and GRA effectiveness results, compatibility environment, in vitro and in vivo component transfer and traceability, content measurability, component-target network degree value, and target binding activity in the spectral-effect analysis of Example 1 were respectively labeled as P1 to P6, and the normalization results are shown in Table 4. Based on the data of the six dimensions, a six-dimensional "spider web" model was established, a radar chart was drawn using R language, and the regression area was calculated. The six-dimensional characteristic network results of the "spider web" model and the histogram of the regression area rankings are as Figure 4 shown.

[0040] Normalization formula: X' = (X - X min ) / (X max - X min ).

[0041] Table 4 Normalized data of candidate bioactive components in the six-dimensional "spider web" model

[0042] As can be seen from Table 4 and Figure 4 it can be known that the bioactive compounds with higher regression areas are salvianolic acid B, tanshinone IIA, dihydrotanshinone I, rosmarinic acid, isoquercitrin, lithospermic acid, cryptotanshinone, ursolic acid, procyanidin B2, hyperoside, etc. The above bioactive compounds are the key substances for the Salvia miltiorrhiza - Crataegus pinnatifida pair to exert myocardial protection. Based on the 6 - dimensional "spider web" model, the present invention screened out the pharmacodynamic substances of 10 Salvia miltiorrhiza - Crataegus pinnatifida pairs, which is of great significance for further exploring the potential action mechanism of the Salvia miltiorrhiza - Crataegus pinnatifida pair in the treatment of ischemic heart disease.

[0043] The above - described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely represents the selected embodiments of the present invention. All other embodiments obtained by relevant deductions and substitutions made by those of ordinary skill in the art under the condition of the inventive concept of the present invention without creative work fall within the scope of protection of the present invention.

Claims

1. A method for screening effective substances of traditional Chinese medicine based on a 6-dimensional cobweb model, characterized in that, Including: Construct a 6-dimensional feature network; Normalize the data of each dimension in the 6-dimensional feature network to obtain quantization values; Link the quantization values of the same candidate active ingredient in different dimensions respectively to construct regression regions of different candidate active ingredients; Sort the areas of the regression regions, screen the candidate active ingredients with regression region areas higher than 0.30 to obtain the traditional Chinese medicine effective substances; The candidate active ingredients are composed of danshensu, chlorogenic acid, procyanidin B2, epicatechin, hyperoside, isoquercitrin, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid, and ursolic acid; The 6-dimensional feature network is composed of the effectiveness dimension, compatibility environment dimension, transmission and traceability dimension, content measurability dimension, network pharmacology degree value dimension, and molecular docking target binding activity dimension of the candidate active ingredients; The traditional Chinese medicine in the traditional Chinese medicine effective substances is the Danshen-Hawthorn herb pair, which is prepared by compounding Danshen and Hawthorn herbs according to the mass ratio of Danshen:Hawthorn = 1:

3.

2. The method for screening active ingredients of traditional Chinese medicine based on a 6-dimensional cobweb model according to claim 1, wherein The data acquisition method for the effectiveness dimension includes: Based on orthogonal partial least squares, associate the levels of myocardial injury markers with the peak areas of the common peaks in the Danshen-Hawthorn herb pair to obtain the variable projection importance values of the compounds corresponding to the common peaks and anti-myocardial injury activity; Based on grey relational analysis, associate the levels of myocardial injury markers with the peak areas of the common peaks in the Danshen-Hawthorn herb pair to obtain the correlation degree values of the compounds corresponding to the common peaks and anti-myocardial injury activity; Integrate the variable projection importance values and correlation degree values of the compounds corresponding to the common peaks and anti-myocardial injury activity to obtain the data of the effectiveness dimension.

3. The method for screening effective substances of traditional Chinese medicine based on a 6D cobweb model according to claim 2, characterized in that, The myocardial injury markers are lactate dehydrogenase and creatine kinase isoenzyme.

4. The method for screening effective substances of traditional Chinese medicine based on a 6D cobweb model according to claim 1, characterized in that The data acquisition method for the compatibility environment dimension includes: According to the compatibility ratio of the Danshen-Hawthorn herb pair, set the weight of the candidate active ingredients only present in Danshen to 25, the weight of the candidate active ingredients only present in Hawthorn to 75, and the weight of the candidate active ingredients present in both Danshen and Hawthorn to 100; Normalize the weights of the candidate active ingredients to obtain the data of the compatibility environment dimension.

5. The screening method of traditional Chinese medicine effective substances based on a 6-dimensional cobweb model according to claim 1, characterized in that The data acquisition method for the transmission and traceability dimension includes: Analyze the distribution of the candidate active ingredients in in vitro samples, serum samples, and heart tissue samples; Set the weight of the distribution of the candidate active ingredients in in vitro samples to 20, the weight of the distribution of the candidate active ingredients in serum samples to 80, and set the weight of the distribution of the candidate active ingredients in heart tissue samples to 100, and obtain the data of the transmission and traceability dimension after normalization.

6. The method for screening active ingredients of traditional Chinese medicine based on a 6D cobweb model according to claim 1, wherein, The data acquisition method for the content measurability dimension includes: Determine the contents of the candidate active ingredients in different batches of the Danshen-Hawthorn herb pair; Normalize the contents of the candidate active ingredients to obtain the data of the content measurability dimension.

7. The method for screening effective substances of traditional Chinese medicine based on a 6-dimensional cobweb model according to claim 1, wherein, The data acquisition method for the network pharmacology degree value dimension includes: Visualize the candidate active ingredients and their action targets to construct a component-target network; The Analyzer tool is applied to calculate the network topology parameters to obtain the data of the network pharmacology degree value dimension.

8. The method for screening the effective substances of traditional Chinese medicine based on the 6D cobweb model according to claim 1, wherein, The method for obtaining the data of the binding activity dimension between the candidate active ingredients and the targets includes: Performing molecular docking on the candidate active ingredients and the core targets, and calculating the binding free energy between the candidate active ingredients and the core targets; Normalizing the binding free energy to obtain the data of the binding activity dimension; The core targets consist of STAT3, SRC, TP53, JUN, and AKT1.

9. The pharmacodynamic substances screened by the screening method of traditional Chinese medicine pharmacodynamic substances based on the 6D cobweb model according to any one of claims 1 to 8, characterized in that, It consists of salvianolic acid B, tanshinone IIA, dihydrotanshinone I, rosmarinic acid, isoquercitrin, lithospermic acid, cryptotanshinone, ursolic acid, procyanidin B2, and hyperoside.

10. Use of the pharmacodynamic substance according to claim 9 in establishing the quality evaluation standard for the Salvia miltiorrhiza - Crataegus pinnatifida pair.

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