Compositions for regulating platelet aggregation, screening methods targeting platelet-derived growth factor receptor α, and applications thereof

Abstract

The present invention relates to the technical field of traditional Chinese medicine chemistry, and specifically relates to a composition for regulating platelet aggregation, a screening method targeting platelet-derived growth factor receptor α and its application. The composition can effectively inhibit platelet aggregation. Experimental results prove that the composition has obvious curative effects in regulating platelet aggregation. The dosage form of the composition is tablets, capsules, syrups, oral liquids, injections, etc. The combined compounds of the present invention have the advantages of significant curative effects, no contraindications and adverse reactions, and can be used for a long time.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine chemistry, and particularly relates to a composition for regulating platelet aggregation, a screening method targeting platelet-derived growth factor receptor α, and applications thereof. Background Art

[0002] Platelet-derived growth factor receptor α (PDGFRA) is a transmembrane single-chain glycoprotein belonging to the tyrosine kinase receptor, which plays an important role in angiogenesis, cell division promotion, etc. It mainly exists in cells regulated by platelet-derived growth factor, such as endothelial cells, smooth muscle cells, fibroblasts, glomerular cells, tumor cells, etc. It plays a role in platelet activation, secretion of platelet granule agonists, and thrombin-induced platelet aggregation. PDGFRA antagonists can block the occurrence of these effects, have great application prospects, have a protective effect on cerebral ischemia and cerebral inflammatory injury, and have good therapeutic and preventive effects on PDGFRA-induced cardiovascular diseases. Therefore, it is of great significance to search for and develop new, efficient, safe, low-toxic, and convenient combined compounds with the effect of regulating platelet aggregation based on PDGFRA. Summary of the Invention

[0003] Aiming at the deficiencies of the prior art, the present invention provides a composition for regulating platelet aggregation targeting platelet-derived growth factor receptor α (PDGFRA).

[0004] Another object of the present invention is to provide a method for screening this composition targeting platelet-derived growth factor receptor α (PDGFRA) through virtual screening molecular docking, or in combination with in-vitro experiment verification.

[0005] The object of the present invention is achieved in the following manner:

[0006] A composition for regulating platelet aggregation, the composition comprising the following components: geraniol, cyasterone, kakkalide, hemerocallin, and formononetin.

[0007] Preferably, the composition for regulating platelet aggregation is 2-4 parts by weight of geraniol, 0.7-1.9 parts by weight of cyasterone, 1.1-3.1 parts by weight of kakkalide, 0.7-1.3 parts by weight of hemerocallin, and 0.8-1.6 parts by weight of formononetin.

[0008] Most preferably, the composition for regulating platelet aggregation is: 2 parts by weight of geraniol, 0.7 parts by weight of cyasterone, 1.1 parts by weight of kakkalide, 0.7 parts by weight of hemerocallin, and 0.8 parts by weight of formononetin.

[0009] A method for screening the above composition for regulating platelet aggregation using platelet-derived growth factor receptor α (PDGFRA) as a target, comprising the following steps:

[0010] 1) Obtaining the three-dimensional structure of platelet-derived growth factor receptor α (PDGFRA) protein;

[0011] 2) Using molecular docking software, determining the active site region of the platelet-derived growth factor receptor α (PDGFRA) protein structure with Sunitinib as the center;

[0012] 3) Performing computer virtual screening on the small molecule ligands of five traditional Chinese medicines, Bupleuri Radix, Cyathulae Radix, Puerariae Flos, Hemerocallis Root, and Puerariae Lobatae Radix, and the platelet-derived growth factor receptor α (PDGFRA) protein. According to the set active site region, using molecular docking software, docking each small molecule ligand with the active site region of the platelet-derived growth factor receptor α (PDGFRA), and recording the docking score data;

[0013] 4) Sorting according to the docking score results in step 3) (for each traditional Chinese medicine, selecting the small molecule ligand with the highest docking score), and determining the composition that can regulate platelet aggregation: geraniol, cyasterone, kakkalide, hemerocallin, and formononetin.

[0014] In the above method, before obtaining the three-dimensional structure of the platelet-derived growth factor receptor α (PDGFRA) protein, searching the public database for the effective chemical components of five traditional Chinese medicines, Bupleuri Radix, Cyathulae Radix, Puerariae Flos, Hemerocallis Root, and Puerariae Lobatae Radix, a total of 264 chemical component information was obtained, including 83 for Bupleuri Radix, 36 for Cyathulae Radix, 104 for Puerariae Flos, 24 for Hemerocallis Root, and 17 for Puerariae Lobatae Radix. Each compound was pre-treated and optimized using the MacroModel small molecule ligand pre-treatment optimization module.

[0015] In the above method, each compound was pre-treated and optimized using the MacroModel small molecule ligand pre-treatment optimization module, and the energy threshold was set to 0.5 kJ·nm -1 ·mol -1 .

[0016] In step 1) of the above method, the three-dimensional structure of platelet-derived growth factor receptor α (PDGFRA) protein is specifically obtained from the RCSB PDB protein database with the PDB ID of 5GRN. This protein has a total of 356 amino acid residues, and its construction method is X-RAY DIFFRACTION, with a resolution of:

[0017] In step 1) of the above method, after obtaining the three-dimensional structure of platelet-derived growth factor receptor α (PDGFRA) protein, the constructed protein can be subjected to model validation and molecular dynamics to evaluate the structural rationality.

[0018] In step 2) of the above method, the specific method for determining the active site region of the platelet-derived growth factor receptor α (PDGFRA) protein structure with Sunitinib as the center is as follows: The platelet-derived growth factor receptor α (PDGFRA) protein and Sunitinib are respectively added to the molecular docking Maestro system for pre-processing such as adding hydrogen atoms, deleting water molecules, and processing heavy atoms. The receptor grid generation (i.e., active pocket generation) panel under the glide docking module is used to determine the active site region of the platelet-derived growth factor receptor α (PDGFRA) protein structure with Sunitinib as the center.

[0019] The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α (PDGFRA) as a target specifically includes the following steps:

[0020] 1) Search for the effective chemical components of five traditional Chinese medicines, Bupleurum chinense, Cyathula officinalis, Flos Puerariae, Hemerocallis fulva L., and Pueraria lobata, from public databases, and a total of 264 chemical component information is obtained, including 83 for Bupleurum chinense, 36 for Cyathula officinalis, 104 for Flos Puerariae, 24 for Hemerocallis fulva L., and 17 for Pueraria lobata. Each compound is pre-processed and optimized using the MacroModel small molecule ligand pre-processing optimization module.

[0021] 2) Obtain the three-dimensional structure of platelet-derived growth factor receptor α (PDGFRA) protein from the RCSB PDB protein database with the PDB ID of 5GRN. This protein has a total of 356 amino acid residues, and its construction method is X-RAY DIFFRACTION, with a resolution of:

[0022] 3) Add the platelet-derived growth factor receptor α (PDGFRA) protein and sunitinib obtained in step 2) into the molecular docking Maestro system respectively, perform preprocessing such as adding hydrogen atoms, deleting water molecules, and processing heavy atoms. Use the receptor grid generation panel under the glide docking module (i.e., active pocket generation) to determine the active site region of the PDGFRA protein structure with sunitinib as the center; import the 264 compound molecules of the four traditional Chinese medicines obtained in step 1) into the molecular docking Maestro system, and set the distance between the PDGFRA protein and the compound molecules (i.e., the small molecule ligands in the previous text) to 1.0 / 0.8; select standard precision in terms of accuracy, that is: introduce the 264 compound molecules of the five traditional Chinese medicines, namely bupleurum root, cyathula root, flos puerariae, hemerocallis root, and pueraria root, into the active site region of the PDGFRA protein structure respectively, and conduct virtual screening with the PDGFRA protein. Perform molecular docking between each compound and the active site region of the PDGFRA protein, and record the docking score data; sort according to the docking scores to determine the composition that can regulate platelet aggregation: geraniol, cyasterone, kakkalide, hemerocallin, and formononetin.

[0023] 4) Conduct bioactivity verification through in-vivo pharmacological experiments.

[0024] The present invention conducts the following specific in-vivo pharmacological experiment verification on the method for screening the composition that regulates platelet aggregation with platelet-derived growth factor receptor α (PDGFRA) as the target:

[0025] 1) Take ICR mice weighing 18 - 22 g, administer the corresponding drugs respectively, give the drugs by tail vein injection, and continuously administer for 7 days. After the last administration, take blood from the mouse eyeballs within 1.5 - 2.5 hours, anticoagulate with sodium heparin, and centrifuge to obtain the supernatant to get the plasma containing the drug.

[0026] 2) Select a mouse platelet-derived growth factor receptor α (PDGFRA) enzyme-linked immunosorbent assay (ELISA) kit, use the double antibody sandwich method to measure the level of mouse platelet-derived growth factor receptor α (PDGFRA) in the specimen, measure the absorbance (OD value) with an enzyme-labeled instrument, and calculate the concentration of platelet-derived growth factor receptor α (PDGFRA) in the plasma containing the drug of the mouse through the standard curve.

[0027] 3) Divide the difference between each measured value and that of the blank control group by the measured value of the blank control group to obtain the inhibitory rate of platelet-derived growth factor receptor α (PDGFRA), and evaluate the pharmacodynamic characteristics of the combination of five compounds, namely geraniol, cyasterone, kakkalide, hemerocallin, and formononetin, in inhibiting PDGFRA.

[0028] The composition screened by the method of the present invention can be prepared into a preparation with pharmaceutically acceptable excipients, and the dosage form can be tablets, capsules, syrups, oral liquids, injections, etc. The traditional Chinese medicine composition of the present invention has the advantages of no obvious toxic and side effects, significant curative effect, no contraindications and adverse reactions, and can be used for a long time.

[0029] The last object of the present invention is to disclose the application of the aforementioned composition for regulating platelet aggregation in the preparation of drugs for anti-platelet aggregation or regulating platelet aggregation.

[0030] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0031] (1) The present invention screens platelet aggregation inhibitors with geraniol, cyasterone, kakkalide, hemerocallin, and formononetin as the composition. The compounds are derived from traditional Chinese medicinal materials, have no obvious toxic and side effects, significant curative effect, no contraindications and adverse reactions, and can be used for a long time.

[0032] (2) The present invention provides a method for screening the traditional Chinese medicine composition with anti-platelet aggregation effect by combining virtual screening molecular docking method and in-vitro experiment verification with PDGFRA as the target, which has high precision and accurate and reliable results.

[0033] (3) The present invention first uses virtual screening molecular docking technology to screen the most excellent compound monomers for the PDGFRA target from five traditional Chinese medicinal materials, namely Bupleuri Radix, Cyathulae Radix, Puerariae Flos, Hemerocallidis Radix, and Puerariae Lobatae Radix, and then combines and proportions the five screened monomers, which has an obvious effect on regulating platelet aggregation and shows synergistic effects. The drug ratio used in the composition of the present invention is verified by a large number of experiments. Too low or too high dosage of any drug will affect the efficacy of regulating platelet aggregation. Detailed implementation manners

[0034] The present invention will be further explained and illustrated through the following examples:

[0035] Example 1:

[0036] (1) Search for the active chemical components of five traditional Chinese medicines, namely Bupleuri Radix, Cyathulae Radix, Puerariae Flos, Hemerocallis Root, and Puerariae Lobatae Radix, from the database (TCMSP Traditional Chinese Medicine Systems Pharmacology Platform). A total of 264 chemical component information were obtained, including 83 for Bupleuri Radix (the names of the included compounds are shown in Table 1), 36 for Cyathulae Radix (the names of the included compounds are shown in Table 2), 104 for Puerariae Flos (the names of the included compounds are shown in Table 3), 24 for Hemerocallis Root (the names of the included compounds are shown in Table 4), and 17 for Puerariae Lobatae Radix (the names of the included compounds are shown in Table 5). For each compound, use the MacroModel small molecule ligand preprocessing and optimization module to preprocess and optimize the active component database (this library consists of 264 chemical component information), and the energy threshold is set to 0.5 kJ·nm -1 ·mol -1 . The MacroModel small molecule ligand preprocessing and optimization module converts the two-dimensional planar structure of the compound into a three-dimensional structure and performs energy optimization preprocessing;

[0037] (2) Obtain the three-dimensional structure of the PDGFRA protein from the RCSB PDB protein database. Its PDB ID is 5GRN. This protein has a total of 356 amino acid residues, and its construction method is X-ray diffraction (X-RAY DIFFRACTION), with a resolution of:

[0038] (3) Import the PDGFRA protein and Sunitinib obtained in step (2) into the molecular docking Maestro system respectively. Perform preprocessing such as adding hydrogen atoms (since there are no hydrogen atoms in the protein structure, so add hydrogen atoms in batches), deleting water molecules, and processing heavy atoms. Use the receptor grid generation panel under the glide docking module to determine the active site region of the PDGFRA protein structure with Sunitinib as the center; set the distance between the PDGFRA protein and the compound molecule to 1.0 / 0.8, select "standard precision" in terms of accuracy, perform molecular docking of Sunitinib with the active site region of PDGFRA, and record the total docking score. The docking score of Sunitinib and PDGFRA is -9.0401. Use this score as a threshold to judge the interaction effect of the five traditional Chinese medicinal materials and PDGFRA. Introduce the 264 small molecule compounds obtained in step (1) into the active site region of the PDGFRA protein structure respectively, and perform virtual screening with the PDGFRA protein, and record the docking score data (the docking score data is mainly composed of the lipophilicity, hydrogen bonds, metal ligands, inappropriate bond rotations, and steric repulsion between the small molecule compound and the PDGFRA protein. The docking score data is calculated by the molecular docking system according to the built-in logic of the molecular docking system and then directly output). The results show that geraniol in Bupleuri Radix (see Table 1 for details, docking score -9.792), cyasterone in Cyathulae Radix (see Table 2 for details, docking score -9.652), kakkalide in Puerariae Flos (see Table 3 for details, docking score -9.942), hemerocallin in Hemerocallis Root (see Table 4 for details, docking score -9.663), and formononetin in Puerariae Lobatae Radix (see Table 5 for details, docking score -9.509) have the highest scores after docking with PDGFRA (that is, the highest absolute value of the score), above the threshold set by Sunitinib (that is, compare the absolute value of each docking score with the absolute value of the docking score of Sunitinib). Therefore, select 5 compounds, namely geraniol, cyasterone, kakkalide, hemerocallin, and formononetin, for the combination ratio of compounds regulating platelet aggregation.

[0039] Table 1 Virtual screening results of Bupleuri Radix and platelet-derived growth factor receptor alpha (PDGFRA)

[0040]

[0041]

[0042]

[0043] Table 2 Virtual screening results of Cyathula officinalis Kuan and platelet-derived growth factor receptor α (PDGFRA)

[0044]

[0045]

[0046] Table 3 Molecular docking results of Pueraria lobata flower and platelet-derived growth factor receptor α (PDGFRA)

[0047]

[0048]

[0049]

[0050]

[0051] Table 4 Molecular docking results of Hemerocallis fulva root and platelet-derived growth factor receptor α (PDGFRA)

[0052]

[0053]

[0054] Table 5 Molecular docking results of Pueraria lobata root and platelet-derived growth factor receptor α (PDGFRA)

[0055]

[0056] The following in-vivo experiments were conducted to verify the method for screening compositions that can regulate platelet aggregation using PDGFRA as a target:

[0057] (1) 140 ICR mice, weighing 18 - 22 g, were produced and distributed by Shanghai Jieshijie Experimental Animal Co., Ltd. They were randomly divided into 28 groups of 5 mice each. Each group was given the monomers or compositions of 5 compounds, namely geraniol, cyasterone, kakkalide, hemerocallin, and formononetin, corresponding to groups 1 - 28 in Table 6 (the types and doses of drugs given to each group are shown in Table 6). Each drug was dissolved in physiological saline according to the corresponding dose in advance and administered by tail vein injection for 7 consecutive days. At 1.5 - 2.5 hours after the last dose, the eyeballs of the ICR mice were removed to collect blood, anticoagulated with sodium heparin, and centrifuged at 5000 revolutions per minute to obtain the drug-containing plasma supernatant for standby.

[0058] Table 6 Detection results of PDGFRA enzyme-linked immunosorbent assay kit

[0059]

[0060]

[0061] A: geraniol; B: cyasterone; C: kakkalide; D: hemerocallin; E: formononetin. * P < 0.05, compared with the blank

[0062] control group, there was a statistically significant difference; a P < 0.05, compared with Group 4 (A: 4 mg / kg -1 ), there was a statistically significant difference; b P < 0.05, compared with Group 22 (C: 1.1 mg / kg -1 ; D: 0.8 mg / kg -1 ), there was a statistically significant difference; c P < 0.05, compared with Group 25 (D: 0.7 mg / kg -1 ; B: 0.7 mg / kg -1 ; C: 1.1 mg / kg -1 ), there was a statistically significant difference. d P < 0.05, compared with Group 27 (A: 2 mg / kg -1 ; B: 0.7 mg / kg -1 ; C: 1.1 mg / kg -1 ; D: 0.7 mg / kg -1 ), there was a statistically significant difference.

[0063] (2) Select the mouse platelet-derived growth factor receptor α (PDGFRA) enzyme-linked immunosorbent assay (ELISA) kit provided by Shanghai Sig Biotech Co., Ltd. Refer to the instruction manual and use the double antibody sandwich method to measure the level of mouse PDGFRA in the specimen (i.e., plasma containing drugs). Measure the absorbance (OD value) at a wavelength of 450 nm using a Synergy 2 multimode microplate reader from BioTek Instruments, Inc., USA, and calculate the concentration of mouse PDGFRA in the sample (i.e., plasma containing drugs) through the standard curve.

[0064] (3) Divide the difference between the measured value of each plasma containing drugs and the blank control group by the measured value of the blank control group to obtain the PDGFRA inhibition rate. The detailed results are shown in Table 6 (the data of 5 mice in each group together constitute the inhibition rate column in Table 6), where the measured value is the measured concentration value of mouse PDGFRA in the plasma containing drugs. The results showed that in Group 28 (geraniol 2 mg / kg -1 , cyasterone 0.7 mg / kg -1 , kakkalide 1.1 mg / kg -1, hemerocallin 0.7 mg / kg -1 and formononetin 0.8 mg / kg -1 ) Compared with the blank group, the highest values of the low, medium, and high dose groups of the 5 single compounds (Group 4), the highest values of the two combinations of the 5 compounds (Group 22), the highest values of the three combinations of the 5 compounds (Group 25), and the highest values of the four combinations of the 5 compounds (Group 27), there was statistical significance, indicating that the combination of the 5 compounds geraniol, cyasterone, kakkalide, hemerocallin, and formononetin of the present invention had a good inhibition rate on PDGFRA. Therefore, the combination of the 5 compounds geraniol, cyasterone, kakkalide, hemerocallin, and formononetin could regulate platelet aggregation (manifested as inhibition).

[0065] Example 2

[0066] geraniol 2 parts by weight, cyasterone 0.7 parts by weight, kakkalide 1.1 parts by weight, hemerocallin 0.7 parts by weight, and formononetin 0.8 parts by weight;

[0067] Mix the above components with the granule excipients and prepare granules according to the conventional preparation method.

[0068] Example 3

[0069] geraniol 2 parts by weight, cyasterone 0.7 parts by weight, kakkalide 1.1 parts by weight, hemerocallin 0.7 parts by weight, and formononetin 0.8 parts by weight;

[0070] Mix the above components with the tablet excipients and press into tablets according to the conventional preparation method.

[0071] Example 4

[0072] geraniol 2 parts by weight, cyasterone 0.7 parts by weight, kakkalide 1.1 parts by weight, hemerocallin 0.7 parts by weight, and formononetin 0.8 parts by weight;

[0073] Mix the above components and prepare oral liquid according to the conventional oral liquid preparation method.

[0074] The basic principles, main features and advantages of the present invention have been shown and described above. Those skilled in the art should understand that the above embodiments do not limit the present invention in any form. Any technical solutions obtained by means of equivalent replacement or equivalent transformation shall fall within the protection scope of the present invention.

Claims

1. A composition for regulating platelet aggregation, characterized in that: The composition comprises the following components: Geraniol, cyasterone, kakkalide, hemerocallin and formononetin.

2. The composition for regulating platelet aggregation according to claim 1, characterized in that: The composition: 2-4 parts by weight of geraniol, 0.7-1.9 parts by weight of cyasterone, 1.1-3.1 parts by weight of kakkalide, 0.7-1.3 parts by weight of hemerocallin and 0.8-1.6 parts by weight of formononetin.

3. A method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target, characterized in that: The following steps are involved: 1) Obtaining the three-dimensional structure of platelet-derived growth factor receptor α (PDGFRA) protein; 2) Using molecular docking software, we determined the active site region of the platelet-derived growth factor receptor α (PDGFRA) protein structure with sunitinib as the center; 3) Computer virtual screening was performed on the small molecule ligands of five Chinese medicinal materials, namely, Bupleurum chinense, Cyathula officinalis, Pueraria lobata, Hemerocallis radix and Pueraria lobata, and the platelet-derived growth factor receptor α (PDGFRA) protein. According to the set active site region, molecular docking software was used to perform molecular docking on each small molecule ligand and the platelet-derived growth factor receptor α (PDGFRA) active site region, and the docking score data was recorded; 4) According to the docking score results of step 3), the composition that can regulate platelet aggregation is determined: geraniol, cyasterone, kakkalide, hemerocallin and formononetin.

4. The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target according to claim 3, characterized in that: Before obtaining the three-dimensional structure of platelet-derived growth factor receptor α (PDGFRA) protein, the effective chemical components of five traditional Chinese medicinal materials, namely Bupleurum, Cyathula, Pueraria, Hemerocallis and Pueraria, were searched from the database. A total of 264 chemical components were obtained, including 83 from Bupleurum, 36 from Cyathula, 104 from Pueraria, 24 from Hemerocallis and 17 from Pueraria. The MacroModel small molecule ligand preprocessing optimization module was applied to each compound for preprocessing optimization.

5. The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target according to claim 4, characterized in that: MacroModel small molecule ligand pretreatment optimization module was applied to each compound for pretreatment optimization, and the energy threshold was set to 0.5 kJ×nm -1 ×mol -1 .

6. The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target according to claim 3, characterized in that: In step 1), the three-dimensional structure of the platelet-derived growth factor receptor α (PDGFRA) protein was obtained in the following way: obtained through the RCSB PDB protein database, its PDB ID is 5GRN, the protein has a total of 356 amino acid residues, and its construction method is X-ray diffraction, with a resolution of 1.77 Å.

7. The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target according to claim 3, characterized in that: The specific method for determining the active site region of the platelet-derived growth factor receptor alpha protein (PDGFRA) structure with sunitinib as the center in step 2) is as follows: the platelet-derived growth factor receptor alpha (PDGFRA) protein and sunitinib are respectively added to the molecular docking Maestro system, and hydrogen atoms are added, water molecules are deleted, and heavy atoms are processed before processing. The active pocket generation panel under the glide docking module is used to determine the active site region of the platelet-derived growth factor receptor alpha protein (PDGFRA) structure with sunitinib as the center.

8. The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target according to claim 3, characterized in that: The following steps are involved: (1) The effective chemical components of five traditional Chinese medicines, namely, Bupleurum chinense, Cyathula officinalis, Pueraria lobata, Hemerocallis radix and Pueraria lobata, were searched from the database. A total of 264 chemical components were obtained, including 83 from Bupleurum chinense, 36 from Cyathula officinalis, 104 from Pueraria lobata, 24 from Hemerocallis radix and 17 from Pueraria lobata. The MacroModel small molecule ligand pretreatment optimization module was used to optimize the pretreatment of each compound. (2) The structure of platelet-derived growth factor receptor α (PDGFRA) protein was obtained through the RCSB PDB protein database. Its PDB ID is 5GRN. The protein has 356 amino acid residues and was constructed by X-ray diffraction with a resolution of 1.77Å. (3) The platelet-derived growth factor receptor α protein and sunitinib obtained in step (2) were added to the molecular docking Maestro system respectively, and hydrogen atoms were added, water molecules were deleted, and heavy atoms were processed. The active pocket generation panel under the glide docking module was used to determine the active site region of the platelet-derived growth factor receptor α (PDGFRA) protein structure with sunitinib as the center; 264 compound molecules of the five traditional Chinese medicines obtained in step (1) were introduced into the molecular docking Maestro system , the distance between the platelet-derived growth factor receptor α (PDGFRA) protein and the compound molecules was set to 1.0 / 0.8; that is, 264 compound molecules from five Chinese medicinal materials, namely, Bupleurum, Cyathula, Pueraria, Hemerocallis and Pueraria, were introduced into the active site region of the platelet-derived growth factor receptor α (PDGFRA) protein structure, and virtual screening was performed with the platelet-derived growth factor receptor α (PDGFRA) protein. Each compound was molecularly docked with the active site region of the platelet-derived growth factor receptor α (PDGFRA), and the docking score data was recorded; The docking scores were ranked and the combination that can regulate platelet aggregation was identified: geraniol, cyasterone, kakkalide, hemerocallin and formononetin; (4) Verify biological activity through physical pharmacology experiments.

9. The method for screening a composition for regulating platelet aggregation using platelet-derived growth factor receptor α as a target according to claim 8, characterized in that: The pharmacological experimental steps are as follows: (1) ICR mice weighing 18-22 g were given the corresponding drugs by tail vein injection for 7 consecutive days. Blood was collected from the mouse eyeballs within 1.5-2.5 hours after the last administration, and anticoagulated with heparin sodium. The supernatant was centrifuged to obtain drug-containing plasma; (2) The mouse PDGFRA enzyme-linked immunosorbent assay kit was used to determine the level of mouse platelet-derived growth factor receptor (PDGFRA) in the specimen using the double antibody sandwich method. The absorbance was measured using an enzyme-labeled instrument, and the concentration of platelet-derived growth factor receptor α (PDGFRA) in the drug-containing plasma of the mouse was calculated using the standard curve. (3) The difference between each measured value and the blank control group was divided by the measured value of the blank control group to obtain the platelet-derived growth factor receptor α (PDGFRA) inhibition rate, so as to evaluate the pharmacodynamic characteristics of the five compound combinations in inhibiting platelet-derived growth factor receptor α (PDGFRA).

10. Use of the platelet aggregation regulating composition according to claim 1 or 2 in the preparation of drugs for resisting platelet aggregation or regulating platelet aggregation.

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