A method for screening effective substances in traditional Chinese medicine based on a 6-dimensional spider web model

Abstract

The present invention discloses a method for screening effective substances of traditional Chinese medicine based on a 6-dimensional spider web model, belonging to the field of medical technology. Based on the results of UHPLC analysis, the present invention uses danshensu, chlorogenic acid, procyanidin B2, epicatechin, hyperoside, isoquercetin, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid, and ursolic acid as the "specific" candidate effective ingredients of the Danshen-Hawthorn medicinal pair. Based on the data of the candidate effective ingredients in six dimensions, namely, effectiveness dimension, compatibility environment dimension, transmission and traceability dimension, content measurability dimension, network pharmacology degree value dimension, and molecular docking target binding activity dimension, a "spider web" model is constructed to obtain the key effective substances of the Danshen-Hawthorn medicinal pair. The present invention provides theoretical support for revealing the compatibility theory of the Danshen-Hawthorn medicinal pair, formulating its quality evaluation standards, and clinical application of the Danshen-Hawthorn medicinal pair.

Description

Technical Field

[0001] The invention belongs to the field of medical technology and relates to a method for screening effective substances of traditional Chinese medicine based on a 6-dimensional spider web model. Background Art

[0002] The Danshen-Haoshan herbal pair, a classic prescription from Shi Jinmo's "Duiyao," consists of two Chinese herbs, Danshen and Hawthorn. It is a classic clinical combination for the treatment of ischemic heart disease based on the TCM theory of promoting blood circulation and removing blood stasis. Danshen, slightly cold in nature and bitter in flavor, enters the heart and liver meridians, and has the effects of promoting blood circulation and removing blood stasis, cooling blood and eliminating carbuncles, and clearing the heart and relieving restlessness. Hawthorn, slightly warm in nature and sour and sweet in flavor, enters the spleen, stomach, and liver meridians, and has the effects of promoting digestion and eliminating accumulation, promoting qi and dispersing blood stasis, and clearing turbidity and reducing lipids. The combination of the two herbs produces a synergistic effect, significantly improving the symptoms of chest pain caused by blood stasis, and holds significant clinical value in the treatment of cardiovascular disease.

[0003] The pharmacological substances of traditional Chinese medicine (TCM) refer to the chemical components or groups of components in TCM and its compound formulas that can characterize clinical efficacy. Research on these components includes key scientific issues such as identification of chemical components or groups of components, analysis of in vivo metabolic processes, and confirmation of target sites. Traditional research models, which primarily focus on the isolation and purification of single components combined with activity screening, have difficulty fully elucidating the holistic "multi-component, multi-target, and multi-pathway" characteristics of TCM. Therefore, establishing a research method that aligns with the holistic characteristics of TCM and systematically uncovering the synergistic mechanisms of the various components in TCM compound formulas has significant theoretical significance and application value in promoting the modernization of TCM research.

[0004] The spider-web model is a multivariate statistical analysis method based on radar chart principles. It enables comprehensive evaluation of complex systems through the visualization of multidimensional evaluation indicators. Currently, research on the pharmacological basis of the Danshen-Hawthorn combination for myocardial ischemia is insufficient, and the scientific implications and quality control standards for its compatibility urgently need to be clarified. Therefore, exploring and developing a spider-web model-based method for screening effective compounds in traditional Chinese medicine will provide important technical support for clarifying the compatibility patterns of the Danshen-Hawthorn combination, establishing a scientific quality evaluation system, and guiding rational clinical drug use. Summary of the Invention

[0005] To solve the above technical problems, the present invention provides a method for screening effective substances of traditional Chinese medicine based on a 6-dimensional spider web model. Based on the results of UHPLC analysis, the present invention uses danshensu, chlorogenic acid, proanthocyanidin B2, epicatechin, hyperoside, isoquercetin, 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 ingredients. Based on the six dimensions of the candidate effective ingredients, namely, "effectiveness" dimension, "compatibility environment" dimension, "transfer and traceability" dimension, "content measurability" dimension, network pharmacology "degree value" dimension, and molecular docking "target binding activity" dimension, a "spider web" model is constructed to obtain the effective substances of the Danshen-Hawthorn drug pair. The present invention provides theoretical support for revealing the compatibility theory of the Danshen-Hawthorn drug pair, formulating the quality evaluation standard of the Danshen-Hawthorn drug pair, and the clinical application of the Danshen-Hawthorn drug pair.

[0006] To achieve the technical purpose of the present invention, on the one hand, the present invention provides a method for screening effective Chinese medicine substances based on a 6-dimensional spider web model, wherein the Chinese medicine in the effective Chinese medicine substance is a Danshen-Hawthorn drug pair, and the method for screening effective Chinese medicine substances comprises:

[0007] Based on the concept of "five principles" in traditional Chinese medicine quality markers, a 6-dimensional feature network is formed; the data of each dimension in the 6-dimensional feature network is normalized to obtain a quantitative value; the quantitative values ​​of the same candidate efficacy component in different dimensions are linked respectively to construct a regression region of the candidate efficacy component; the areas of the regression regions are sorted, and candidate efficacy components with regression region areas greater than 0.30 are screened to obtain the traditional Chinese medicine efficacy substance;

[0008] Specifically, the candidate active ingredients are composed of danshensu, chlorogenic acid, proanthocyanidin B2, epicatechin, hyperoside, isoquercetin, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid, and ursolic acid;

[0009] Specifically, the six-dimensional feature network consists of the "effectiveness" dimension, "compatibility environment" dimension, "transfer and traceability" dimension, "content measurability" dimension, network pharmacology "degree value" dimension, and molecular docking "target binding activity" dimension of the candidate effective ingredients.

[0010] Furthermore, in the method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model provided by the present invention, the data acquisition method of the "effectiveness" dimension includes:

[0011] Correlating the levels of myocardial injury markers with the peak areas of the common peaks in the Danshen-Hawthorn drug pair based on orthogonal partial least squares method, and obtaining the variable projection importance values ​​of the compounds corresponding to the common peaks and the anti-myocardial injury activity;

[0012] Based on the grey correlation analysis, the myocardial injury marker levels were correlated with the peak areas of the common peaks in the Danshen-Hawthorn drug pair to obtain the correlation values ​​between the compounds corresponding to the common peaks and the anti-myocardial injury activity;

[0013] The variable projection importance values ​​and correlation values ​​of the compounds corresponding to the common peaks and the anti-myocardial injury activity are normalized to obtain the data of the "effectiveness" dimension.

[0014] Specifically, the myocardial injury markers are lactate dehydrogenase and creatine kinase isoenzymes.

[0015] Furthermore, in the method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model provided by the present invention, the data acquisition method of the "compatibility environment" dimension includes:

[0016] The weight of the candidate active ingredients in the Danshen-Hawthorn pair that are only present in Danshen is set to 25, the weight of the candidate active ingredients that are only present in Hawthorn is set to 75, and the weight of the candidate active ingredients that are present in both Danshen and Hawthorn is set to 100;

[0017] The weights of the candidate effective ingredients are normalized to obtain data of the "compatibility environment" dimension.

[0018] Furthermore, in the method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model provided by the present invention, the data acquisition method of the "transfer and traceability" dimension includes:

[0019] Analyzing the distribution of the candidate effective ingredients in in vitro samples, serum samples, and heart tissue samples;

[0020] The weight of the distribution of candidate effective ingredients in in vitro samples was set to 20, the weight of the distribution of candidate effective ingredients in serum samples was set to 80, and the weight of the distribution of candidate effective ingredients in heart tissue samples was set to 100. After normalization, the data of the "transfer and traceability" dimension was obtained.

[0021] Specifically, the present invention uses chromatography-mass spectrometry to analyze and identify the in vitro chemical composition of the Danshen-Crataegus herbal extract, as well as the serum-transferred components and cardiac tissue-distributed components after oral administration. Ten prototype components were identified, present in both serum and cardiac tissue: tanshinone IIA, lithospermic acid and its isomers, rosmarinic acid, salvianolic acid A, quercetin, dimethyl lithospermate, lithospermic acid, salvianolic acid B, dihydrotanshinone I, and cryptotanshinone. These components can be rapidly distributed to cardiac tissue through the bloodstream and exert a myocardial protective effect. Components found only in serum include danshensu, chlorogenic acid, hyperoside, and isoquercitrin.

[0022] Furthermore, in the method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model provided by the present invention, the data acquisition method of the "content measurability" dimension includes:

[0023] The Danshen and Hawthorn medicinal materials purchased from different sources were compounded in a mass ratio of Danshen: Hawthorn = 1:3 to obtain the Danshen-Hawthorn medicinal pair;

[0024] Determining the content of the candidate active ingredient in the Danshen-Haoshan medicinal pair;

[0025] The content of the candidate effective ingredients is normalized to obtain data of the "content measurability" dimension.

[0026] Specifically, the present invention measured the content of candidate active ingredients in 14 batches of Danshen-Hawthorn herb pairs purchased from different manufacturers using a UHPLC method. The average values ​​of 16 candidate active ingredients in each of the 14 batches of herbal extracts were calculated, reflecting the concentration of each ingredient in the Danshen-Hawthorn herb pair. Finally, the data were normalized to provide data for content measurability.

[0027] Furthermore, in the method for screening effective Chinese medicine substances based on the 6-dimensional spider web model provided by the present invention, the data acquisition method of the "degree value" dimension of network pharmacology includes:

[0028] Visualizing the candidate active ingredients and their targets to construct an ingredient-target network;

[0029] The Analyzer tool was used to calculate the network topology parameters and obtain the data of the "degree value" dimension of the network pharmacology.

[0030] Specifically, the present invention predicted 386 relevant targets of the Danshen-Hawthorn drug pair using the TCMSP, Swiss Target Prediction and Batman-Tcm platforms. The candidate effective components and targets were imported into Cytoscape 3.7.2 software for visualization, a "component-target" network was constructed, and the Analyzer tool was used to calculate the network topology parameters to obtain the degree values ​​of the candidate effective components.

[0031] Furthermore, in the method for screening effective substances of traditional Chinese medicine based on the 6-dimensional "spider web" model provided by the present invention, the data acquisition method of the molecular docking "target binding activity" dimension includes:

[0032] Perform molecular docking of the candidate active ingredient with the core target and calculate the binding free energy of the candidate active ingredient with the core target;

[0033] Normalizing the binding free energy to obtain data on the “target binding activity” dimension of molecular docking;

[0034] The core targets consist of STAT3, SRC, TP53, JUN and AKT1.

[0035] On the other hand, the present invention requests protection for the pharmacological substances screened by the above-mentioned method for screening pharmacological substances of traditional Chinese medicine based on the 6-dimensional spider web model, which are composed of salvianolic acid B, tanshinone ⅡA, dihydrotanshinone I, rosmarinic acid, isoquercitrin, lithospermic acid, cryptotanshinone, ursolic acid, proanthocyanidin B2 and hyperoside.

[0036] In addition, the present invention seeks to protect the use of the above-mentioned medicinal substances in establishing the quality evaluation standard of the Danshen-Hawthorn medicinal pair.

[0037] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0038] (1) The present invention provides a method for screening effective substances of traditional Chinese medicine based on a 6-dimensional spider web model. A 6-dimensional feature network is formed based on the concept of the "five principles" in traditional Chinese medicine quality markers, specifically including the candidate effective ingredients in the "effectiveness" dimension, "compatibility environment" dimension, "transfer and traceability" dimension, "content measurability" dimension, network pharmacology "degree value" dimension, and molecular docking "target binding activity" dimension. The measurement data of each dimension are subjected to minimum-maximum normalization processing to eliminate the analysis bias caused by big data fluctuations. Finally, a radar chart is drawn using R language, and the regression area is calculated. The regression area is ranked, and the candidate effective ingredients with a regression area greater than 0.30 are screened out as the traditional Chinese medicine effective substances.

[0039] (2) The present invention screened out the active substances in the Danshen-Hawthorn pair through a method for screening active substances of traditional Chinese medicine based on a 6-dimensional spider web model, and found that the active substances are composed of salvianolic acid B, tanshinone ⅡA, dihydrotanshinone I, rosmarinic acid, isoquercitrin, lithospermic acid, cryptotanshinone, ursolic acid, proanthocyanidin B2, and hyperoside. The active substances screened out by the present invention facilitate the formulation of quality evaluation standards for the Danshen-Hawthorn pair, ensuring the stability of the efficacy and controllability of the Danshen-Hawthorn pair in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention.

[0041] Figure 1 This is the UHPLC chromatogram of the Danshen-Hawthorn drug pair at a detection wavelength of 280 nm. Figure 1 A in the figure is the UHPLC chromatogram of the mixed reference substance at a detection wavelength of 280 nm; Figure 1Figure B is the UHPLC chromatogram of the Danshen-Hawthorn drug pair at a detection wavelength of 280 nm. The accompanying figures are labeled 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.

[0042] Figure 2 This is the UHPLC chromatogram of Danshen-Crataegus pinnatifida medicine at a detection wavelength of 335 nm. Figure 2 A in the figure is the UHPLC chromatogram of the mixed reference substance at a detection wavelength of 335 nm; Figure 2 Figure B is the UHPLC chromatogram of the Salvia miltiorrhiza-Crataegus pinnatifida drug pair at a detection wavelength of 335 nm. The accompanying figures are labeled as follows: Peak 11: Chlorogenic acid; Peak 12: Hyperoside; Peak 13: Isoquercitrin; Peak 14: Rosmarinic acid; Peak 15: Lithospermic acid.

[0043] Figure 3 The UHPLC chromatogram of Danshen-Crataegus fructus at a detection wavelength of 210 nm. Figure 3 A in the figure is the UHPLC chromatogram of the mixed reference substance at a detection wavelength of 210 nm; Figure 3 Figure B is the UHPLC chromatogram of the Salvia miltiorrhiza-crataegi drug pair at a detection wavelength of 210 nm. The accompanying figures are labeled as follows: Peak 16: oleanolic acid; Peak 17: ursolic acid.

[0044] Figure 4 This is a screening diagram for the active substances of the Salvia miltiorrhiza-crataegi drug pair based on the "spider web" model. Figure 4 A in the figure is the histogram of the regression area ranking of the “spider web” model; Figure 4 B in the figure is the 6-dimensional feature network result of the “spider web” model. DETAILED DESCRIPTION

[0045] The technical solutions of the present invention are described below with reference to the following examples. However, the present invention is not limited to the following examples. The experimental methods and detection methods described in each example are conventional methods unless otherwise specified; the reagents and materials described are commercially available unless otherwise specified.

[0046] Example 1

[0047] This embodiment provides the construction of each dimension of the 6-dimensional feature network in the "spider web" model.

[0048] The present invention forms a 6-dimensional feature network based on the concept of the "five principles" in traditional Chinese medicine quality markers. The candidate effective ingredients are determined by the "specificity" of the ingredients. The 6-dimensional feature network specifically includes: the "effectiveness" dimension of the candidate effective ingredients, the "compatibility environment" dimension, the "transfer 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 "effectiveness" dimension is evaluated by integrating the VIP values ​​of the compounds obtained by the two spectrum-effect analysis methods of OPLS and GRA, and the correlation values ​​between the compounds and the serum markers of myocardial injury, LDH and CK-MB; the "compatibility environment" dimension is evaluated by the compatibility ratio of drug pairs; the "transfer and traceability" dimension is evaluated by detecting the distribution of candidate effective ingredients in vitro and in vivo (rat plasma, rat heart); the "measurability" dimension of content is evaluated by measuring the average content of candidate effective ingredients in extracts of multiple batches of drug 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 uses molecular docking to calculate the binding energy of candidate effective ingredients with core targets for evaluation.

[0049] 1. Screening of candidate active ingredients based on their “uniqueness”

[0050] In the early stage, UHPLC was used to analyze the chemical components of the extracts of the drug pair Danshen-Hawthorn (the mass ratio of Danshen to Hawthorn was 1:1) at different wavelengths. Figures 1 to 3 After comparison with reference substances, 17 chemical components were ultimately identified, including: Peak 1: Salvianolic acid B, Peak 2: 5-hydroxymethylfurfural, Peak 3: Danshensu, Peak 4: Proanthocyanidin 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, and Peak 17: Ursolic acid. Peak 2 was identified as 5-hydroxymethylfurfural, an organic compound typically formed by the dehydration of reducing sugars during heat-related preparation processes. Although 5-HMF has been reported to have other activities, such as antioxidant, anti-allergic, anti-hyperuricemic, and anti-cancer, it is not a compound specific to Danshen and Crataegus pinnatifida and has been detected in many herbs, honey, and sugary foods. Therefore, the other 16 components were selected as candidate active ingredients of the Danshen-Crataegus pinnatifida medicinal pair.

[0051] 2. Evaluation based on the dimensions of “effectiveness” and “compatibility environment”

[0052] Spectrum-effect relationship analysis was initially performed using SIMCA 14.1 software and SPSS PRO online software. Serum levels of myocardial injury markers (lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB)) were correlated with the peak areas of the common peaks in the corresponding samples in mice treated with different Danshen-Hawthorn herb combinations (Danshen:Hawthorn mass ratios of 1:0, 0:1, 2:1, 1:1, 1:2, 1:3, 1:4, and 1:5, respectively). Orthogonal partial least squares (OPLS) modeling and gray relational analysis (GRA) were used for spectrum-effect relationship analysis, and the optimal Danshen-Hawthorn herb combination ratio of 1:3 was identified. The compound variable projected importance (VIP) values ​​obtained from the OPLS model and the correlation values ​​between the compounds and LDH and CK-MB obtained from the GRA model (Table 1) were normalized and used as the "effectiveness" dimension to evaluate compound efficacy. Different drugs have different compatibility ratios, and their efficacy and material basis will also vary. Starting from the perspective of the "compatibility environment" of the drug pair, the weights of the Salvia miltiorrhiza and Crataegus pinnatifida candidates are set to 25 and 75 respectively, and the common components of the two are set to 100. Finally, the weights of each component are normalized.

[0053] Table 1 Spectrum-effect analysis of the activity of Danshen-Hawthorn medicine against myocardial injury

[0054]

[0055] 3. Evaluation based on the dimension of “transmission and traceability”

[0056] The components of the Danshen-Hawthorn pair need to enter the blood or heart to exert their cardioprotective effects. Therefore, the candidate compounds were evaluated in terms of "transmission and traceability" of entry into the blood and heart. In the early stage, the comprehensive use of chromatography-mass spectrometry technology was used to analyze and identify the in vitro chemical components of the Danshen-Hawthorn pair extract, as well as the serum migration components and cardiac tissue distribution components after oral administration. A total of 10 prototype components were found to exist in both serum and cardiac 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 quickly distributed to cardiac tissue through the blood circulation to exert myocardial protective effects. Components that only exist in serum were also found, including tanshinone, chlorogenic acid, hyperoside, and isoquercitrin. The weights of candidates that can enter the heart and blood are set to 100 and 80 respectively, and the weight of candidates that can only be detected in in vitro samples is set to 20. Finally, the weights of each component are normalized.

[0057] 4. Evaluation based on the “measurability” dimension of content

[0058] Measurability of the content of traditional Chinese medicine ingredients is a crucial factor in ensuring the quality and evaluation of medicinal materials. Therefore, from the perspective of measurability, average content was selected as an evaluation metric. Previously, a methodological validation was conducted using UHPLC to determine the content of candidate active ingredients in 14 batches of Danshen-Hawthorn herb pairs (Danshen and Hawthorn herbs from the same manufacturer were paired). The average values ​​of the 16 candidate active ingredients in each of the 14 batches of extracts were calculated (Table 2). This value reflects the concentration of each ingredient in the Danshen-Hawthorn herb pair. Finally, the data were normalized to serve as the "content dimension."

[0059] Table 2 Contents of candidate active ingredients in 14 batches of Danshen-Hawthorn medicinal pairs

[0060]

[0061] 5. Evaluation of the “degree value” dimension of the “component-target” network

[0062] In the early stage, 386 relevant targets of the Danshen-Hawthorn drug pair were predicted by TCMSP, Swiss Target Prediction and Batman-Tcm platforms. The candidate effective ingredients and targets were imported into Cytoscape 3.7.2 software for visualization, and a "component-target" network was constructed. The network topology parameters were calculated using the Analyzer tool to obtain the degree values ​​of the candidate effective ingredients. The larger the degree value, the higher the importance of the ingredient. The degree values ​​of the candidate effective ingredients were ursolic acid 54, salvianolic acid B 51, tanshinone ⅡA 41, tanshinone I 39, rosmarinic acid 34, dihydrotanshinone I 32, cryptotanshinone 31, lithospermic acid 26, oleanolic acid 18, salvianolic acid A 12, tanshinone 11, hyperoside 10, procyanidin B2 7, epicatechin 7, isoquercetin 4, and chlorogenic acid 2. Finally, the degree values ​​of the candidate efficacy components are normalized as the "degree value" dimension.

[0063] 6. Evaluation of the “target binding activity” dimension

[0064] Molecular docking of candidate active ingredients with the top five core disease targets screened by network pharmacology (STAT3, SRC, TP53, JUN, and AKT1, with STAT3 being a key target involved in myocardial ischemia) was performed to evaluate the specific binding activity of the candidate active ingredients with the disease targets. Two-dimensional structural formulas of the candidate active ingredients were obtained from the PubChem database, and three-dimensional molecular models were constructed using Chem3D software and exported as mol2 files. Crystal structures of the core targets were then downloaded from the PDB database and pre-processed using PyMOL software (removing non-critical components such as water molecules and phosphate groups). Molecular structure pre-processing was performed using AutoDockTools 1.5.6, including format conversion (pdb→pdbqt) and active site identification. Rigid docking was performed using Autodock Vina to calculate binding free energies, while flexible docking was performed using Discovery Studio 2019 to obtain the LibDockScore for comprehensive evaluation of ligand-receptor interactions. Based on the Autodock Vina calculation results (Table 3), complexes with binding energies ≤ −5.0 kcal / mol were considered stable binding systems. Three-dimensional conformational visualization was performed using PyMOL software, and key binding sites were analyzed using two-dimensional interaction maps. Finally, the "target binding activity" dimension was obtained by normalizing the combined binding energies of the candidate active ingredients with the five targets.

[0065] Table 3 Binding energy of candidate active ingredients and core disease targets

[0066]

[0067] Example 2

[0068] This embodiment provides the establishment of a "spider web" model.

[0069] The six dimensions of OPLS and GRA effectiveness results, compatibility environment, in vivo and in vitro component delivery and traceability, content measurability, component-target network degree value, and target binding activity in the spectrum-effect analysis of Example 1 are marked as P1 to P6, and the normalized results are shown in Table 4. Based on the data of the six dimensions, a six-dimensional "spider web" model was established, and a radar chart was drawn using R language, and the regression area was calculated. The histogram of the six-dimensional characteristic network results and regression area ranking of the "spider web" model is shown in Figure 4. Figure 4 shown.

[0070] Normalization formula:

[0071] X'=(XX min ) / (X max -X min ).

[0072] Table 4 Normalized data of candidate efficacy components in the 6-dimensional “spider web” model

[0073]

[0074] From Table 4 and Figure 4 The active compounds with the highest regression areas are, in order, salvianolic acid B, tanshinone IIA, dihydrotanshinone I, rosmarinic acid, isoquercetin, lithospermic acid, cryptotanshinone, ursolic acid, procyanidin B2, and hyperoside. These active compounds are key components of the Danshen-Hawthorn pair in exerting myocardial protection. This study, based on a six-dimensional "spider web" model, screened 10 active components of the Danshen-Hawthorn pair, which is of great significance for further investigation into the potential mechanism of action of the Danshen-Hawthorn pair in treating ischemic heart disease.

[0075] The embodiments described above are some of the embodiments of the present invention, rather than all of them. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but rather merely represents selected embodiments of the present invention. All other embodiments obtained without creative effort and through deduction and substitution by a person of ordinary skill in the art based on the concept of the present invention are 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 spider web model, characterized in that: include: Construct a 6-dimensional feature network; Normalizing the data of each dimension in the 6-dimensional feature network to obtain a quantized value; Link the quantitative values ​​of the same candidate efficacy component in different dimensions respectively and construct regression regions of different candidate efficacy components; sorting the areas of the regression regions, screening candidate active ingredients with regression area greater than 0.30, and obtaining the Chinese medicinal active substances; The candidate active ingredients are composed of danshensu, chlorogenic acid, proanthocyanidin B2, epicatechin, hyperoside, isoquercetin, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, oleanolic acid, and ursolic acid; The six-dimensional feature network consists of the effectiveness dimension of the candidate active ingredients, the compatibility environment dimension, the delivery and traceability dimension, the content measurability dimension, the network pharmacology value dimension, and the molecular docking target binding activity dimension; The Chinese medicine in the Chinese medicinal active substance is a pair of salvia miltiorrhiza and hawthorn, which is obtained by compounding salvia miltiorrhiza and hawthorn according to a mass ratio of salvia miltiorrhiza to hawthorn = 1:3; The data acquisition method of the effectiveness dimension includes: Correlating the levels of myocardial injury markers with the peak areas of the common peaks in the Danshen-Hawthorn drug pair based on orthogonal partial least squares method, and obtaining the variable projection importance values ​​of the compounds corresponding to the common peaks and the anti-myocardial injury activity; Based on the grey correlation analysis, the myocardial injury marker levels were correlated with the peak areas of the common peaks in the Danshen-Hawthorn drug pair to obtain the correlation values ​​between the compounds corresponding to the common peaks and the anti-myocardial injury activity; Integrating the variable projection importance values ​​and correlation values ​​of the compounds corresponding to the common peaks and the anti-myocardial injury activity to obtain data of the effectiveness dimension; The data acquisition methods for the transmission and traceability dimensions include: Analyzing the distribution of the candidate effective ingredients in in vitro samples, serum samples, and heart tissue samples; The weight of the candidate efficacy component distribution in in vitro samples was set to 20, the weight of the candidate efficacy component distribution in serum samples was set to 80, and the weight of the candidate efficacy component distribution in heart tissue samples was set to 100. After normalization, the data of the transfer and traceability dimensions were obtained; The data acquisition method of the network pharmacology degree value dimension includes: Visualizing the candidate active ingredients and their targets to construct an ingredient-target network; Calculating network topology parameters using the Analyzer tool to obtain data on the network pharmacology degree value dimension; Methods for obtaining data on the binding activity dimension of the candidate active ingredient and the target include: Perform molecular docking of the candidate active ingredient with the core target and calculate the binding free energy of the candidate active ingredient with the core target; Normalizing the binding free energy to obtain data on the binding activity dimension; The core targets consist of STAT3, SRC, TP53, JUN and AKT1.

2. The method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model according to claim 1, characterized in that: The myocardial injury markers are lactate dehydrogenase and creatine kinase isoenzymes.

3. The method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model according to claim 1, characterized in that: The data acquisition method of the compatibility environment dimension includes: According to the compatibility ratio of the Danshen-Hawthorn drug pair, the weight of the candidate active ingredients present only in Danshen is set to 25, the weight of the candidate active ingredients present only in Hawthorn is set to 75, and the weight of the candidate active ingredients present in both Danshen and Hawthorn is set to 100; The weights of the candidate effective ingredients are normalized to obtain data of the compatibility environment dimension.

4. The method for screening effective substances of traditional Chinese medicine based on the 6-dimensional spider web model according to claim 1, characterized in that: Methods for obtaining data on the content measurability dimension include: Determining the content of candidate active ingredients in different batches of the Danshen-Haoshan medicinal pair; The content of the candidate effective ingredients is normalized to obtain data on the content measurability dimension.

5. The pharmacological substance obtained by screening the pharmacological substance screening method of traditional Chinese medicine based on the 6-dimensional spider web model according to any one of claims 1 to 4, characterized in that: It is composed of salvianolic acid B, tanshinone ⅡA, dihydrotanshinone Ⅰ, rosmarinic acid, isoquercetin, lithospermic acid, cryptotanshinone, ursolic acid, proanthocyanidin B2 and hyperoside.

6. Use of the pharmacological substance according to claim 5 in establishing a quality evaluation standard for the Danshen-Haoshan medicinal pair.

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