Pharmaceutical composition for colorectal adenoma and application thereof

Abstract

The invention relates to a pharmaceutical composition for colorectal adenoma and application thereof. The pharmaceutical composition comprises at least one of the following components: a compound 1 and / or a pharmaceutically acceptable salt thereof; compound 2 and / or a pharmaceutically acceptable salt thereof; compound 3 and / or a pharmaceutically acceptable salt thereof; and a compound 4 and / or a pharmaceutically acceptable salt thereof, which can be used for treating and / or preventing diseases or conditions mediated by TNF-alpha. Target spots and active ingredients of the traditional Chinese medicine compound are determined through a network pharmacology research method, and the method has important application significance for guiding research and development of the traditional Chinese medicine compound.

Description

[0001] This application claims priority to an earlier application filed on May 17, 2024, with the China National Intellectual Property Administration, patent application number 2024106186169, entitled "Pharmaceutical Composition for Colorectal Adenoma and Its Application Therein". The entire contents of the earlier application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of pharmaceuticals, specifically relating to a pharmaceutical composition for colorectal adenoma and its application. Background Technology

[0003] Tubular adenomas are the most common precancerous lesions of colorectal cancer, and precise prevention and treatment are key to reducing the incidence of colorectal cancer. The high recurrence rate after resection of colorectal adenoma lesions presents a significant challenge in diagnosis and treatment. Currently, patients can only monitor and prevent recurrence through colonoscopy, which is costly and cannot be widely applied to the general population. There is an urgent need to explore effective recurrence prevention strategies. Traditional Chinese medicine (TCM) has the advantage of reducing postoperative recurrence rates and improving patient symptoms in the treatment of colorectal adenomas, but treatment protocols lack standardization. Furthermore, the components of specialized compound preparations are complex, and their mechanisms of action are unclear. Therefore, it is crucial to clarify the drug targets and mechanisms of action of TCM specialized compound preparations for the prevention of colorectal adenoma recurrence, further identify their advantageous populations, and determine the biomarkers and intervention pathways for the clinical application of these specialized compound preparations.

[0004] Current research suggests that the detection rate of colorectal adenomas in the first year after resection is 36%–61%, raising awareness of the importance of preventing adenoma recurrence. Much current research focuses on biomarkers for screening, such as peripheral blood DNA, peripheral blood inflammatory factors, and fecal DNA, but their diagnostic efficacy for adenomas is not yet optimal. Drug treatments for preventing recurrence often focus on traditional Chinese medicine. For example, Huchzermeyer et al. proposed that changes in lipid and bile acid metabolism after lactulose intake can prevent colorectal adenomas to some extent, but convincing clinical relevance studies and exploration of specific mechanisms are lacking. Currently, biomarkers for screening colorectal adenomas include multi-target fecal FIT-DNA, circulating blood methylated Septin9 DNA detection, fecal PKM2 protein, and microbial biomarkers. However, these often lack large-scale clinical trials, are expensive, and have limited practical application convenience. Therefore, further development of drugs to prevent adenoma recurrence is needed.

[0005] Compound berberine hydrochloride tablets (also known as berberine tablets) are mainly composed of four traditional Chinese medicines: berberine hydrochloride, costus root, evodia fruit, and white peony root. They have the effects of clearing heat and drying dampness, promoting qi circulation and relieving pain, and stopping dysentery and diarrhea. They are commonly used for damp-heat in the large intestine, bloody or white dysentery, tenesmus or sudden diarrhea, and burning sensation in the anus; also for enteritis and dysentery with the above symptoms. Berberine, also known as berberine, has the effects of clearing heat and detoxifying, purging fire and drying dampness, and is used to treat damp-heat fullness, vomiting and diarrhea, high fever and thirst. Costus root promotes qi circulation and relieves pain, strengthens the spleen and promotes digestion, and is used for chest and abdominal distension and pain, tenesmus after diarrhea, and indigestion. Evodia fruit warms the middle jiao, relieves pain, regulates qi, and dries dampness, and can treat vomiting and diarrhea due to cold in the internal organs, abdominal distension and pain, and early morning diarrhea. White peony root soothes the liver and regulates qi, relieves spasms and pain, and can be used to treat abdominal and chest pain, such as abdominal pain due to deficiency and cold in the middle jiao. Current research indicates that, in addition to its anti-inflammatory effects, compound berberine tablets also have anti-tumor, hypoglycemic, hypolipidemic, and immune-enhancing effects.

[0006] Network pharmacology is an interdisciplinary science that explores the intervention and effects of complex drugs on diseases based on pharmacology, systems biology, big data, and network analytics, thereby further revealing the mechanisms by which drugs exert their effects in the body. Applying network pharmacology to the development of traditional Chinese medicine can overcome the difficulty in elucidating the mechanisms of action in TCM. Based on network pharmacology, the effects and mechanisms of compound berberine hydrochloride tablets in the treatment and prevention of colorectal adenomas can be analyzed and verified, leading to the development of drugs for the treatment and prevention of colorectal adenomas. Summary of the Invention

[0007] To address the above problems, the present invention provides a composition comprising at least one selected from the following: compound 1 and / or a pharmaceutically acceptable salt thereof; compound 2 and / or a pharmaceutically acceptable salt thereof; compound 3 and / or a pharmaceutically acceptable salt thereof; and compound 4 and / or a pharmaceutically acceptable salt thereof;

[0008] Compounds 1, 2, 3, and 4 have the chemical structures shown below:

[0009]

[0010] According to an embodiment of the present invention, the active ingredient in the composition comprises: compound 1 and / or a pharmaceutically acceptable salt thereof; compound 2 and / or a pharmaceutically acceptable salt thereof; compound 3 and / or a pharmaceutically acceptable salt thereof; and compound 4 and / or a pharmaceutically acceptable salt thereof.

[0011] According to an embodiment of the present invention, the active ingredient in the composition comprises the following components: compound 1 and / or a pharmaceutically acceptable salt thereof; compound 2 and / or a pharmaceutically acceptable salt thereof; compound 3 and / or a pharmaceutically acceptable salt thereof; and compound 4 and / or a pharmaceutically acceptable salt thereof.

[0012] According to an embodiment of the present invention, the composition comprises berberine or a pharmaceutically acceptable salt thereof, as well as costus root, evodia fruit and white peony root.

[0013] According to an embodiment of the present invention, the composition comprises berberine, costus root, evodia fruit and white peony root extract.

[0014] According to embodiments of the present invention, the binding energy of the active ingredient in the composition to TNF-α is ≤-5 kcal / mol, for example ≤-6 kcal / mol, preferably -10 kcal / mol to -5 kcal / mol. As examples, the binding energies of the active ingredient in the composition to TNF-α are -9.2 kcal / mol, -6.1 kcal / mol, -9.2 kcal / mol, and -9.5 kcal / mol.

[0015] According to embodiments of the present invention, the active ingredient of the composition does not contain any substances other than berberine, costus root, evodia fruit, and white peony root, except for: compound 1 and / or its pharmaceutically acceptable salt; compound 2 and / or its pharmaceutically acceptable salt; compound 3 and / or its pharmaceutically acceptable salt; and compound 4 and / or its pharmaceutically acceptable salt.

[0016] The present invention also provides a pharmaceutical composition comprising at least one of the above compositions, and one or more pharmaceutically acceptable excipients.

[0017] The present invention also provides the use of at least one of the above-described compositions for the preparation of a medicament for the treatment and / or prevention of diseases or conditions mediated by TNF-α.

[0018] The present invention also provides a method for treating and / or preventing diseases or conditions mediated by TNF-α, the method comprising administering at least one of the above-described compositions to a patient in need.

[0019] According to embodiments of the present invention, the TNF-α-mediated disease or condition is a tumor or tumor recurrence. For example, the tumor is a malignant tumor (such as cancer), such as a malignant solid tumor, preferably a colorectal adenoma (e.g., colorectal adenocarcinoma).

[0020] In the context of this invention, "recurrence" refers to the reappearance or spread of tumor cells (such as tumor cells of colorectal adenoma) that were previously thought to have been eliminated or controlled after treatment.

[0021] According to an embodiment of the invention, the patient in need may be a mammal, such as a human.

[0022] This invention also provides a method for determining the target of a traditional Chinese medicine compound, wherein the method includes the following steps:

[0023] (1) The effective active ingredients and targets of traditional Chinese medicine compound were extracted from the TCM Systems Pharmacology Database and Analysis Platform (TCMSP). The effective active ingredients and targets of each drug in the traditional Chinese medicine compound were screened out under the conditions of oral bioavailability (OB) ≥30% and drug-like properties (DL) ≥0.18.

[0024] (2) Gene search: Genes related to colorectal adenoma were searched through GeneCards, OMIM, PharmGk and DrugBank databases respectively, and the databases were merged to obtain genes related to colorectal adenoma.

[0025] (3) Target analysis: The target IDs of each component in the traditional Chinese medicine compound are converted, and the R language is used to intersect the converted target IDs with the genes in step (2) to obtain the potential targets of the traditional Chinese medicine compound related to colorectal adenoma.

[0026] (4) Network construction: Based on the potential target genes obtained in step (3), a traditional Chinese medicine compound regulatory network and a protein interaction network were constructed using Cytoscape. Protein interaction network analysis yielded 10 core network genes, including TNF, IL6, and TP53.

[0027] (5) Pathway enrichment: KEGG pathway enrichment analysis was performed on genes in the regulatory network of traditional Chinese medicine compound using R language to obtain the pathways enriched with target genes related to colorectal adenoma.

[0028] (6) Double screening: The intersection of the core genes obtained from the protein interaction network analysis in step (4) and the top 10 gene pathways with the significance of the KEGG pathway enrichment analysis in step (5) is used to obtain the target TNF.

[0029] (7) Combined analysis: The 2D structure of small molecule ligands of the components in the traditional Chinese medicine compound was converted into a 3D structure, and water molecules and small molecules were removed from the protein receptor of the drug target. The optimal binding points of small molecule ligands and protein receptors of the components in the traditional Chinese medicine compound were determined.

[0030] (8) Molecular docking of drug targets for colorectal adenoma lesions with effective active ingredients in traditional Chinese medicine compound formulas.

[0031] Optionally, after docking is completed in step (8), the binding energy between the active ingredient molecule and the target site is further detected.

[0032] According to an embodiment of the present invention, the traditional Chinese medicine compound can be compound berberine hydrochloride.

[0033] According to an embodiment of the present invention, the compound berberine hydrochloride contains berberine hydrochloride, costus root, evodia fruit, and white peony root in a weight ratio of (20-40):(100-120):(30-50):(150-170), such as 30:116:40:162.

[0034] Beneficial effects

[0035] The inventors investigated clinical treatment protocols for preventing recurrence of colorectal adenomas and provided evidence-based medicine. Furthermore, they conducted in-depth research on the association between colorectal adenomas and target genes, as well as their binding properties with active ingredients. This research provided a comprehensive TCM-Western medicine intervention protocol for preventing colorectal adenoma recurrence using compound berberine hydrochloride, offering evidence support for the integrated TCM-Western medicine approach to prevent postoperative recurrence of colorectal adenomas. This invention is expected to provide a practical basis for researchers to further develop new drugs, identify effective active ingredients in TCM compound formulas, and provide treatment and / or prevention protocols for colorectal adenomas, or for preventing colorectal adenoma recurrence. Attached Figure Description

[0036] Figure 1A This is a schematic diagram illustrating the intersection of active ingredient targets and disease targets.

[0037] Figure 1B A network diagram illustrating the regulatory mechanisms of traditional Chinese medicine compound prescriptions;

[0038] Figure 1C A protein-protein interaction network diagram of traditional Chinese medicine compound preparations;

[0039] Figure 2A This is a schematic diagram of protein-protein interaction network analysis in traditional Chinese medicine compound preparations.

[0040] Figure 2B This is a schematic diagram of KEGG pathway enrichment analysis;

[0041] Figure 3 The diagram shows the docking of TNF-α with kaempferol, benzoylpaeoniflorin, quercetin, and evodiamine.

[0042] Figure 4 Example 2: Changes in cellular protein levels as shown in the Western Blot experiment;

[0043] Figure 5 The results of the cell heat transfer experiment in Example 2;

[0044] Figure 6A This is a schematic diagram of the mouse model used in the animal experiment of Example 2;

[0045] Figure 6B Example 2: Comparison of mouse body weights in different groups during animal experiments;

[0046] Figure 6C Example 2: Comparison of intestinal length in mice of different groups in animal experiments. Detailed Implementation

[0047] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0048] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0049] Example 1: Target Analysis and Mechanism Study of Compound Berberine Hydrochloride

[0050] 1. Screening of active ingredients and targets of compound berberine hydrochloride: Using network pharmacology, chemical components of four herbs—Huangliansu (berberine), Muxiang (costus root), Wuzhuyu (Evodia fruit), and Baishao (white peony root)—were collected from the TCM Systems Pharmacology Database and Analysis Platform (TCMSP). Active ingredients meeting the criteria of oral bioavailability (OB) ≥ 30% and drug-like properties (DL) ≥ 0.18 were screened. The resulting active ingredients included 14 from berberine, 5 from costus root, 13 from white peony root, and 30 from Evodia fruit, as shown in Table 1.

[0051] Table 1:

[0052]

[0053]

[0054]

[0055] We searched for drug targets using the TCM Systems Pharmacology Database and Analysis Platform (TCMSP), which identified 288 targets for berberine, 43 targets for costus root, 123 targets for white peony root, and 501 targets for evodia fruit.

[0056] 2. Target Screening for Colorectal Adenomas: Genes targeting colorectal adenomas were queried in the GeneCards, OMIM, PharmGkb, and DrugBank databases. GeneCards yielded 4718 results, OMIM 17, DrugBank 46, and PharmGkb 17. Further gene merging of the four databases resulted in 4736 colorectal adenoma-related target genes. The active ingredient target IDs of berberine hydrochloride obtained in step 1 were converted, and R language was used to perform an intersection analysis between the active ingredient target and the colorectal adenoma-related gene files obtained in this step, yielding 143 potential targets of berberine hydrochloride associated with colorectal adenomas, such as... Figure 1A As shown.

[0057] 3. Construction of a compound berberine hydrochloride regulatory network and protein interaction network: Based on the target genes of the above-mentioned colorectal adenoma-related formulas, a compound regulatory network and protein interaction network of traditional Chinese medicine were constructed using Cytoscape, such as... Figure 1B , Figure 1C As shown.

[0058] 4. Protein-Protein Interaction (PPI) Network Analysis and KEGG Pathway Enrichment Analysis: To investigate target-related biological functions, protein-protein interaction (PPI) analysis and KEGG pathway enrichment analysis were conducted. PPI analysis was performed using Cytoscope software, while KEGG pathway enrichment was performed using the dedicated website KEGG:Kyoto Encyclopedia of Genes and Genomes. Both are relatively mature analytical methods. The dedicated website KEGG (Kyoto Encyclopedia of Genes and Genomes) is a database resource containing information on the functions of biological systems. It provides information on genomes, chemical substances, metabolic pathways, diseases, and drugs, covering multiple biological fields, including genomics, metabolomics, and drug discovery. It provides researchers with rich biological information resources for understanding the structure, function, and dynamic changes of biological systems.

[0059] Results of protein interaction network analysis of compound berberine hydrochloride are as follows: Figure 2A As shown, 10 core genes in the regulatory network, including TNF, IL6, and TP53, were obtained. KEGG pathway enrichment analysis was performed on the genes in the compound berberine hydrochloride regulatory network using R language. The KEGG pathway enrichment analysis is shown below. Figure 2BAs shown, the analysis results indicate that target genes related to colorectal adenoma are significantly enriched in pathways such as the "TNF signaling pathway". A double screening was performed on core genes, with the criteria being the intersection of genes identified as core nodes by PPI analysis and the top 10 genes significantly enriched by KEGG pathway analysis. The target TNF was obtained through this secondary screening. Studies have shown that tumor necrosis factor (TNF) is a small molecule protein secreted by macrophages, with a normal serum level of 4.3±2.8 mg / L. TNF-α is mainly produced by macrophages and is the most important pro-inflammatory cytokine involved in cell growth, differentiation, and apoptosis. The levels of TNF-α in the blood and colonic mucosa of patients with proctitis are significantly higher than in the general population, and anti-TNF-α drugs have achieved significant therapeutic effects in the clinical treatment of proctitis. Based on the above results and research, it is suggested that compound berberine hydrochloride may exert its effects through the TNF signaling pathway, which may be a direct drug target of its four herbal components.

[0060] 5. Molecular docking analysis of the binding efficiency of active ingredients to the target: In compound berberine hydrochloride, based on the aforementioned network pharmacology analysis, four active ingredients that directly interact with TNF were identified: kaempferol, paeoniflorin, quercetin, and rutaecarpine. The 2D structures of the small molecule ligands of these four active ingredients were downloaded and converted to 3D structures. Simultaneously, water molecules and small molecules were removed from the protein receptor of the drug target. The optimal binding sites of the small molecule ligands and protein receptors were determined. Finally, molecular docking was completed between the drug target of colorectal adenoma lesions and the active ingredients in compound berberine hydrochloride. The molecular docking results are as follows: Figure 3 As shown. The binding energies of TNF-α with kaempferol, paeoniflorin, quercetin, and rutaecarpine were calculated to be -9.20, -6.10, -9.20, and -9.50 kcal / mol, respectively, indicating that the binding energy is low, the binding efficiency is high, and the binding structure is stable.

[0061] Example 2: Application Study of Compound Berberine Hydrochloride Drug Target Intervention Using In Vivo and In Vitro Experiments

[0062] In vitro validation:

[0063] 1. Inhibitory effect of compound berberine hydrochloride on the proliferation of colon cancer cells.

[0064] The preventive effect of compound berberine hydrochloride on the recurrence of colorectal adenomas was verified using HT 29 and Caco2 colon cancer cell lines (purchased from ATCC), which have a lower malignancy. Different concentrations of compound berberine hydrochloride were used to treat the two cell lines, and cell viability was detected and IC50 was calculated using a CKK8 cell proliferation assay. 50 value.

[0065] Experimental results: After 48 hours of drug treatment, the IC50 value of the HT 29 cell line was [not specified]. 50 The IC50 value for the Caco2 cell line was 54.26 μM. 50 The concentration was 51.99 μM, indicating that the compound berberine hydrochloride had an inhibitory effect on the proliferation of colon cancer cells.

[0066] 2. Western blot analysis of changes in cellular protein levels after treatment with compound berberine hydrochloride.

[0067] Western Blot Experiment: A 100 mM stock solution of compound berberine hydrochloride was prepared using DMSO as the solvent and diluted to 0 μM, 12.5 μM, 25 μM, and 50 μM with the corresponding culture media. HT 29 and Caco 2 were then resuscitated at a concentration of 1 × 10⁻⁶ mg / well. 6 Cells were evenly seeded in 6-well cell culture dishes and incubated overnight. When the cell density reached 80%, fresh medium was added, and different concentrations of pre-prepared compound berberine hydrochloride were added. The dishes were then incubated for 48 hours. After 48 hours of culture, the medium was discarded, and the cells were washed three times with 1×PBS. RIPA lysis buffer and protease inhibitors were added, and the dishes were incubated on ice for 10 minutes. Cells were then collected using a cell scraper, transferred to centrifuge tubes, lysed by sonication, and centrifuged (4°C, 12000 rpm) for 15 minutes. The supernatant was collected. Protein concentration was determined, and the mixture was diluted to 20 μg / 10 μL with DDW and loading buffer. The samples were boiled at 100°C for 10 minutes. Separating and stacking gels were prepared and incubated at room temperature for 30 minutes until completely solidified. Protein samples were loaded at 10 μL / well, electrophoresis was performed at 90 V for 30 min, followed by 120 V for 60 min, and then transferred to a membrane at 300 mA for 90 min. The membrane was then blocked with 5% skim milk for 2 h, and the corresponding primary antibody was added and incubated overnight at 4 °C. β-Tubulin was used as an internal control. The next day, the corresponding secondary antibody was added and incubated at room temperature for 2 h. Finally, protein expression levels were detected using ECL high-sensitivity chemiluminescence buffer.

[0068] Experimental results are as follows Figure 4As shown, the expression levels of the cell-metabolite-related markers NCAD and Vimentin decreased significantly; meanwhile, the expression level of the epithelial-related marker ECAD did not change much, indicating that the drug can maintain cell epitheliality while reversing the stromal nature of tumor cells and effectively improve epithelial-mesenchymal transition (EMT); at the same time, compound berberine hydrochloride can effectively reduce the protein levels of the cell proliferation marker PCNA and the cell stemness marker CD44.

[0069] 3. Cellular thermal transfer assay (CETSA)

[0070] Experimental conditions: HT 29 cells were seeded in 10cm culture dishes. After the cells were almost confluent, two dishes were taken and washed twice with PBS. After trypsin digestion, the cells were added to two 15ml centrifuge tubes and centrifuged at 3000rpm for 3min. The supernatant was discarded, and the cells were gently washed twice more with PBS. For the final wash, 500ul of PBS was added to each tube to resuspend the cells. The drug group was treated with 100μM compound berberine hydrochloride, and the control group was treated with the corresponding volume of carrier solvent DMSO. After thorough mixing, the cells were divided into eight 200ul aliquots. EP tubes, 60 μL each, were incubated at 37°C in a 5% CO2 incubator for 2 h. Cells were then removed, and the drug group and control group were heated for 3 minutes each at corresponding temperature gradients (37°C, 42°C, 47°C, 52°C, 57°C, 62°C, 67°C) using a gradient PCR instrument. Immediately after heating, the cells were placed on ice, and protease inhibitors were added. Cells were lysed using liquid nitrogen freeze-thaw cycles. After centrifugation at 20,000 rcf for 20 min at 4°C, 50 μL of supernatant was collected from each tube and transferred to a new EP tube. Loading buffer was added, and the mixture was thoroughly mixed and boiled at 100°C for 10 min. Finally, the TNF-α protein level was detected using the aforementioned Western blotting method. Experimental results are as follows: Figure 5 As shown, compound berberine hydrochloride binds to TNF-α and enhances the thermal stability of TNF-α protein.

[0071] Animal experiments verified:

[0072] Experimental mice: 6-week-old male C57BL / 6 mice.

[0073] according to Figure 6A The procedure shown is used to construct an AOM / DSS-induced colonic adenoma model.

[0074] Mice were randomly divided into three groups: a normal control group, an AOM / DSS induction group, and an AOM / DSS + compound berberine hydrochloride group, with six mice in each group. Except for the normal control group, all other groups received an intraperitoneal injection of 10 mg / kg AOM, followed by oral administration of 2.5% DSS aqueous solution. The normal control group and the AOM / DSS induction group received sterile saline daily, while the AOM / DSS + compound berberine hydrochloride group received 40 mg / kg compound berberine hydrochloride daily. Mouse weight and symptoms were observed, and intestinal length, adenoma number, and other indicators were compared among the different groups. Experimental results are as follows: Figure 6B , 6C As shown, mice in the AOM / DSS induced group exhibited significant weight loss and a marked shortening of colon length, indicating successful model establishment. Mice in the AOM / DSS + compound berberine hydrochloride group had significantly higher body weight than the AOM / DSS induced group, and their colon length also showed some recovery compared to the AOM / DSS induced group. These results confirm that compound berberine hydrochloride can significantly reduce the severity of AOM / DSS colitis and decrease the incidence of colorectal adenomas.

[0075] The exemplary embodiments of the present invention have been described above. However, the scope of protection of this application is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A composition comprising at least one selected from the group consisting of: compound 1 and / or a pharmaceutically acceptable salt thereof; compound 2 and / or a pharmaceutically acceptable salt thereof; compound 3 and / or a pharmaceutically acceptable salt thereof; and compound 4 and / or a pharmaceutically acceptable salt thereof; in, Compounds 1, 2, 3, and 4 have the chemical structures shown below:

2. The composition according to claim 1, characterized in that, The active ingredient in the composition comprises: compound 1 and / or a pharmaceutically acceptable salt thereof; compound 2 and / or a pharmaceutically acceptable salt thereof; compound 3 and / or a pharmaceutically acceptable salt thereof; and compound 4 and / or a pharmaceutically acceptable salt thereof.

3. The composition according to claim 2, characterized in that, The active ingredient in the composition comprises the following components Composition: Compound 1 and / or its pharmaceutically acceptable salt; Compound 2 and / or its pharmaceutically acceptable salts; Compound 3 and / or its pharmaceutically acceptable salts; and compound 4 and / or its pharmaceutically acceptable salts.

4. The composition according to any one of claims 1-3, characterized in that, The composition comprises berberine or a pharmaceutically acceptable salt thereof, as well as costus root, evodia fruit and white peony root; preferably, the composition comprises extracts of berberine, costus root, evodia fruit and white peony root.

5. The composition according to any one of claims 1-4, characterized in that, The binding energy of the active ingredient in the composition to TNF-α is ≤-5 kcal / mol, for example ≤-6 kcal / mol, preferably -10 kcal / mol to -5 kcal / mol; more preferably, the binding energy of the active ingredient in the composition to TNF-α is -9.2 kcal / mol, -6.1 kcal / mol, -9.2 kcal / mol and -9.5 kcal / mol.

6. The composition according to any one of claims 1-5, characterized in that, The active ingredients of the composition do not contain any substances other than berberine, costus root, evodia fruit, and white peony root, except for the following: compound 1 and / or its pharmaceutically acceptable salts; Compound 2 and / or its pharmaceutically acceptable salt; compound 3 and / or its pharmaceutically acceptable salt; and compound 4 and / or its pharmaceutically acceptable salt.

7. A pharmaceutical composition comprising at least one of the compositions according to any one of claims 1-6, and one or more pharmaceutically acceptable excipients.

8. Use of the composition according to any one of claims 1-6 or the pharmaceutical composition according to claim 7 for the preparation of a medicament for the treatment and / or prevention of a disease or condition mediated by TNF-α, preferably, the disease or condition mediated by TNF-α being a tumor or tumor recurrence; for example, the tumor being a malignant tumor (such as carcinoma), such as a malignant solid tumor, preferably a colorectal adenoma (e.g., colorectal adenocarcinoma).

9. A method for determining the target of a traditional Chinese medicine compound, wherein the method includes the following steps: (1) The effective active ingredients and targets of traditional Chinese medicine compound were extracted from the TCM Systems Pharmacology Database and Analysis Platform (TCMSP). The effective active ingredients and targets of each drug in the traditional Chinese medicine compound were screened out under the conditions of oral bioavailability (OB) ≥30% and drug-like properties (DL) ≥0.

18. (2) Gene search: Genes related to colorectal adenoma were searched through GeneCards, OMIM, PharmGk and DrugBank databases respectively, and the databases were merged to obtain genes related to colorectal adenoma. (3) Target analysis: The target IDs of each component in the traditional Chinese medicine compound are converted, and the R language is used to intersect the converted target IDs with the genes in step (2) to obtain the potential targets of the traditional Chinese medicine compound related to colorectal adenoma. (4) Network construction: Based on the potential target genes obtained in step (3), a traditional Chinese medicine compound regulatory network and a protein interaction network were constructed using Cytoscape. Protein interaction network analysis yielded 10 core network genes, including TNF, IL6, and TP53. (5) Pathway enrichment: KEGG pathway enrichment analysis was performed on genes in the regulatory network of traditional Chinese medicine compound using R language to obtain the pathways enriched with target genes related to colorectal adenoma. (6) Double screening: The intersection of the core genes obtained from the protein interaction network analysis in step (4) and the top 10 gene pathways with the significance of the KEGG pathway enrichment analysis in step (5) is used to obtain the target TNF. (7) Combined analysis: The 2D structure of small molecule ligands of the components in the traditional Chinese medicine compound was converted into a 3D structure, and water molecules and small molecules were removed from the protein receptor of the drug target. The optimal binding points of small molecule ligands and protein receptors of the components in the traditional Chinese medicine compound were determined. (8) Molecular docking of drug targets for colorectal adenoma lesions with effective active ingredients in traditional Chinese medicine compound formulas; Optionally, after docking is completed in step (8), the binding energy between the active ingredient molecule and the target site is further detected.

10. The method according to claim 9, characterized in that, The traditional Chinese medicine compound is compound berberine hydrochloride. Preferably, the weight ratio of berberine hydrochloride, costus root, evodia fruit, and white peony root in the compound berberine hydrochloride is (20-40):(100-120):(30-50):(150-170), such as 30:116:40:162.

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