CN111388484A

CN111388484A - New pharmaceutical application of platypodium

Info

Publication number: CN111388484A
Application number: CN202010371146.2A
Authority: CN
Inventors: 陈孝; 唐欲博; 黄帅; 陈杰; 邹学农
Original assignee: First Affiliated Hospital of Sun Yat Sen University
Current assignee: First Affiliated Hospital of Sun Yat Sen University
Priority date: 2020-05-06
Filing date: 2020-05-06
Publication date: 2020-07-10

Abstract

The invention discloses a new application of spatholobus stem in preparing an EphB4/Ephrin-B2 signal channel inhibitor, and the EphB4/Ephrin-B2 signal channel inhibitor can treat diseases caused by abnormal activation of an EphB4/Ephrin-B2 signal channel, and particularly inhibit the tumor proliferation, migration, angiogenesis and anti-apoptosis effects of non-small cell lung cancer caused by abnormal activation of an EphB4/Ephrin-B2 signal channel.

Description

New pharmaceutical application of platypodium

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of platypodium triquetrum and/or derivatives thereof as a signal pathway inhibitor.

Background

The EphB4/Ephrin-B2 signal path molecule has EphB4(Ephrin type-B receptor4) as a receptor and Ephrin-B2 as a ligand; EphB4 and Ephrin-B2 can be mutually activated, namely, the signal transduction between the receptor and the ligand is bidirectional, and Ephrin-B2 activates EphB4 to be a forward signal, and EphB4 activates Ephrin-B2 to be a reverse signal. The EphB4/Ephrin-B2 signal channel is closely related to nervous system development, physiological pathological angiogenesis and tumor development.

The plastic wistin (Pristein, Pris for short, CAS number 1258-84-0) is a natural quinone methyl triterpene compound separated from various plants of Chenopodiaceae and Hippophae, and has multiple pharmacological activities including antiinflammatory, anti-peroxidation, antioxidant, antimalarial, antifungal and insecticidal activities. In addition, PRIS is reported to be a proteasome inhibitor, but the use of PRIS as an inhibitor of the EphB4/Ephrin-B2 signaling pathway has not been reported.

Disclosure of Invention

Through theoretical analysis and experimental exploration of an inventor, the invention provides a new application of the flat plastic rattan, and particularly relates to an application of the flat plastic rattan and/or derivatives thereof as an EphB4/Ephrin-B2 signal pathway inhibitor, wherein the application comprises the application of the flat plastic rattan in disease mechanism research, the application of the flat plastic rattan in targeted therapy of related diseases and the like.

In some embodiments, the invention may include one or more of the following:

1. use of platypodium effuses and/or derivatives thereof in preparing EphB4/Ephrin-B2 signal pathway inhibitor.

2. The use of item 1, wherein the EphB4/Ephrin-B2 signaling pathway inhibitor comprises an EphB4 inhibitor and/or an Ephrin-B2 inhibitor.

3. The use according to item 1, characterized in that: EphB4/Ephrin-B2 signaling pathway inhibitors are useful for treating diseases in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated.

4. The use according to item 3, wherein: the diseases with abnormal activation of EphB4/Ephrin-B2 signal pathway comprise tumor diseases.

5. The use according to item 4, wherein: the neoplastic disease includes head and neck cancer, ovarian cancer, lung cancer, esophageal cancer, or colorectal cancer.

6. The use according to item 5, wherein: the lung cancer comprises non-small cell lung cancer and/or small cell lung cancer.

7. The use of item 4, wherein the EphB4/Ephrin-B2 signal pathway inhibitor inhibits the EphB4/Ephrin-B2 signal pathway, thereby inhibiting tumor cell proliferation, inhibiting tumor cell migration, inhibiting formation of tumor vascular structures, and promoting tumor cell apoptosis.

8. The use of item 1, wherein the platypodium and/or derivative thereof downregulates the expression level of EphB4 by at least 10%.

9. The use of item 1, wherein the platypodium and/or derivative thereof downregulates the expression level of Ephrin-B2 by at least 10%.

10. The use of item 1, wherein the derivative of wisteria florbunda comprises a pharmaceutically acceptable salt of wisteria florda, a pharmaceutically acceptable ester of wisteria florda, a structural analog of wisteria florda, or a modifier of wisteria florda.

In some embodiments, the invention finds that the patulin can inhibit the EphB4/Ephrin-B2 signaling pathway, and in some embodiments, the patulin dose-dependently inhibits the expression of EphB4 and Ephrin-B2.

In some embodiments, the administration of the target cells of fugetin downregulates the expression level of EphB4 in the target cells by at least 10%, e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100% compared to the absence of fugetin.

In some embodiments, the application of platinoidin downregulates the expression level of Ephrin-B2 in the target cell by at least 10%, such as at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100% compared to the absence of application of platinoidin.

In some embodiments, the use of patulin as an inhibitor of the EphB4/Ephrin-B2 signaling pathway is useful in the treatment of diseases in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated. The disease is improved or relieved by inhibiting the EphB4/Ephrin-B2 signal channel or reducing the molecular expression level of the EphB4/Ephrin-B2 signal channel, so that the abnormally activated EphB4/Ephrin-B2 signal channel is restored to normal level or is inhibited.

In some embodiments, a disease in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated is a neoplastic disease, including benign tumors and malignant tumors, which are cancers. In some embodiments, the disease in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated is head and neck cancer, ovarian cancer, lung cancer, esophageal cancer, or colorectal cancer. In some embodiments, the disease in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated is non-small cell lung cancer and/or small cell lung cancer. In some embodiments, the disease in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated is non-small cell lung cancer. In some embodiments, the ampelopsin is as an EphB4/Ephrin-B2 signal pathway inhibitor, through inhibiting EphB4/Ephrin-B2 signal pathway, thereby inhibiting tumor cell proliferation, inhibiting tumor cell migration, inhibiting tumor cell vascular structure formation, and promoting tumor cell apoptosis.

In some embodiments, the modification of the patulin is a modification of β cryptoxanthin, which does not alter its substance as an EphB4/Ephrin-B2 signaling pathway inhibitor.

In some embodiments, the model of non-small cell lung cancer is a non-small cell lung cancer condition reprogrammed cell, abbreviated CR L C, that lacks primary tumor heterogeneity compared to traditional non-small cell lung cancer cell lines, CR L C is derived from patient non-small cell lung cancer tissue, is closer to clinical levels, and is effective in identifying tumor therapeutics.

The invention has the beneficial effects that:

the invention provides a new application of the flat plastic rattan in preparing an EphB4/Ephrin-B2 signal channel inhibitor for the first time, and the EphB4/Ephrin-B2 signal channel inhibitor can treat diseases caused by abnormal activation of an EphB4/Ephrin-B2 signal channel, and particularly inhibit the tumor proliferation, migration, angiogenesis and anti-apoptosis effects of non-small cell lung cancer caused by abnormal activation of an EphB4/Ephrin-B2 signal channel.

Drawings

FIG. 1 shows that the expression levels of EphB4 and Ephrin-B2 were analyzed by immunoblot analysis after CR L C was treated with 0, 2, 4, and 8. mu.M PRIS for 24 h;

FIG. 2 location of EphB4 expression by immunofluorescence staining after CR L C was treated with 0, 2, 4, 8 μ M PRIS for 24 h;

FIG. 3 PRIS (4. mu.M) and/or NVP-BHG71 (0.1. mu.M) treated CR L C, immunoblot analysis EphB4, Ephrin-B2;

FIG. 4 measurement of mRNA and protein expression levels of EphB4 48 hours after transfection of CR L C with either EphB 4-specific siRNA or EphB 4-non-specific siRNA (Scr siRNA);

FIG. 5 PRIS and/or EphB4 siRNA treated CR L C and immunoblotted to analyze the expression levels of EphB4, Ephrin-B2;

FIG. 6 depicts mRNA and protein expression levels of EphB4 in BEAS-2B cells and CR L C from different patients, 3T3-J2 being feeder cells, as one of the CR L C controls;

FIG. 7 PRIS (4. mu.M) and/or NVP-BHG71 (0.1. mu.M) treatment of CR L C to determine cell proliferation potency;

FIG. 8 determination of cell proliferation potency (panel A) by PRIS and/or EphB4 siRNA treatment of CR L C, number of migrating cells in 10 random fields (panel B), lumen formation potency in vitro (panel C);

FIG. 9 shows the apoptosis and caspase activity and levels after PRIS treatment of CR L C, the detection results of the apoptosis kit (panel A), the determination results of caspase activity (panel B), the immunoblot analysis of cytochrome C and caspase activation levels (panel C), the cell viability levels after different caspase inhibitors (10. mu.M) and PRIS treatment of CR L C (panel D);

FIG. 10 immunoblot analysis of cytochrome C, activated caspase-3, activated caspase-9, Bax, Bcl-2 (Panel A) with or without PRIS addition to CR L C transfected with EphB4 siRNA (Panel B), calculation of Bax/Bcl-2 (Panel B);

FIG. 11 characterization of conditionally reprogrammed non-small cell lung carcinoma cells (CR L C) morphology of CR L C at different growth stages (Panel A), representative immunofluorescence images of CR L C showing phenotype status of CK7 and CEA, DNA staining of nuclei with Hoechst 33342 (Panel B), immunohistochemical staining of CR L C to detect CK7, CEA and TTF-1 (Panel C), Western blot analysis of CR L C, 3T3-J2 and non-small cell lung carcinoma SPCA-1 cell lines CK7, CEA and TTF-1 (Panel D), STR analysis of CR L C (Panel E);

in the above figures, immunoblots were referenced to GAPDH and data were expressed as mean ± SD, indicating P <0.05, P <0.01, ns indicating no significant difference.

Detailed Description

The present invention will be more readily understood by the following examples, taken in conjunction with the general description of the invention, which are intended to illustrate embodiments of certain aspects of the invention, and are not intended to limit the invention. It should be noted that the experimental methods or experimental conditions used in the experimental examples were carried out according to the conventional methods or manufacturer's instructions without specific instructions, and the materials and reagents used in the experimental examples were commercially available without specific instructions.

Example 1 PRIS inhibition of the EphB4/Ephrin-B2 Signal pathway

(1) CR L C cells were treated with various concentrations of PRIS (0, 2, 4, 8. mu.M) for 24 hours and examined by Western blotting, as shown in FIG. 1, which inhibited the expression of EphB4/Ephrin-B2 signaling pathway molecules EphB4 and Ephrin-B2 in a dose-dependent relationship, and immunofluorescent staining, as shown in FIG. 2, which inhibited the nuclear expression of EphB4 and the nuclear expression level of EphB 4.

(2) The results of Western blot analysis of CR L C cells treated with PRIS and/or NVP-BHG712 (0.1. mu.M) significantly inhibited the expression of EphB4 and Ephrin-B2, and the EphB4/Ephrin-B2 signaling pathway was inhibited, whereas the inhibition of EphB4/Ephrin-B2 was not enhanced by treating CR L C cells with PRIS and NVP-BHG712 simultaneously, as compared with NVP-BHG712 alone, thus demonstrating that EphB4 autophosphorylation blocks further signaling, while PRIS inhibits EphB4/Ephrin-B2 signaling.

(3) EphB4 siRNA can silence EphB4 expression in CR L C, and mRNA analysis and Western blotting results are shown in FIG. 4. when CR L C cells are transfected with PRIS and/or EphB4 siRNA, Western blotting results are shown in FIG. 5, and when CR L C cells are treated with PRIS and EphB4 siRNA at the same time, the inhibition of EphB4/Ephrin-B2 is not enhanced as compared with EphB4 siRNA alone, thus it can be shown that EphB4 silencing blocks further signal transduction, while PRIS inhibits EphB4/Ephrin-B2 signal transduction.

In summary, PRIS is a potent EphB4/Ephrin-B2 signaling pathway inhibitor that inhibits EphB4 and/or Ephrin-B2; especially in non-small cell lung cancer.

Example 2 PRIS as EphB4/Ephrin-B2 Signal pathway inhibitors inhibits cell proliferation, cell migration, and vascular Structure formation

(1) According to the analysis of CR L C cells and normal human bronchial epithelial cells BEAS-2B, mRNA analysis and Western blotting results are shown in figure 6, and compared with normal human bronchial epithelial cells, EphB4 in CR L C is highly expressed, which indicates that EphB4/Ephrin-B2 signal channel activation abnormality exists in non-small cell lung cancer.

(2) The results of cell proliferation assays using PRIS and/or NVP-BHG712 treated CR L C cells are shown in FIG. 7, where PRIS and NVP-BHG712 (0.1. mu.M) both exert the effect of inhibiting CR L C cell proliferation through EphB4/Ephrin-B2 signaling pathway inhibition.

(3) The results of the detection of cell proliferation, cell migration and in vitro lumen formation ability of CR L C cells transfected with PRIS and/or EphB4 siRNA are shown in FIG. 8, and both PRIS and EphB4 siRNA exert the effect of inhibiting CR L C cell proliferation, cell migration and in vitro angiogenesis through EphB4/Ephrin-B2 signal channel inhibition.

In conclusion, PRIS, as a potent EphB4/Ephrin-B2 signaling pathway inhibitor, could inhibit tumor cell proliferation, cell migration and in vitro angiogenesis in abnormal activation of EphB4/Ephrin-B2 signaling pathway, especially in non-small cell lung cancer.

Example 3 PRIS promotes apoptosis by inhibiting the EphB4/Ephrin-B2 signaling pathway.

(1) The results of apoptosis measurements using PRIS as EphB4/Ephrin-B2 signaling pathway inhibitors on CR L C cells are shown in FIG. 9, where PRIS significantly promoted CR L C apoptosis (FIG. 9A), where PRIS significantly increased the activity of caspase-9, caspase-3 and caspase-4 (FIG. 9B), as measured by caspase activity, as measured by dose-dependent increase in the expression of activated caspase-9, activated caspase-3 and activated caspase-4 and cytochrome C (FIG. 9C), and where caspase inhibitors include Z-L EHD-FMK for caspase-9 inhibition, Z-DEVD-FMK for caspase-3 inhibition, Ac-L EVD-CHO for caspase-4 inhibition, as shown in FIG. 9D, where PRIS reversed apoptosis promotion, and PRIS was seen to promote apoptosis by activated caspase.

(2) The results of detecting apoptosis-related factors by transfecting CR L C cells with PRIS and/or EphB4 siRNA are shown in FIG. 10, EphB4 siRNA significantly increases cytochrome C release, increases the levels of activated caspase-3 and activated caspase-9, increases the expression ratio of Bax/Bcl-2, and thus it can be seen that the inhibition of EphB4 on EphB4/Ephrin-B2 signal channel to promote apoptosis, and the promotion of apoptosis by treating CR L C cells with PRIS and EphB4 siRNA is not enhanced compared with that by using EphB4 alone, thus it can be seen that PRIS also promotes apoptosis by inhibiting EphB4/Ephrin-B2 signal channel.

In conclusion, PRIS, as a potent EphB4/Ephrin-B2 signaling pathway inhibitor, could promote the activation of aberrant tumor cell apoptosis in EphB4/Ephrin-B2 signaling pathways, especially in non-small cell lung cancer.

Materials and methods

Tissue origin: the tissue sample of the patient with the non-small cell lung cancer is obtained from lung resection or ultrasound-guided puncture biopsy of the first hospital affiliated to Zhongshan university.

Cell isolation and culture fresh non-small cell lung carcinoma tissue collected was minced, primary cells were isolated by shaking with collagenase, hyaluronidase and dispase (StemCell Technologies) for 3h at 37 deg.C, and the primary cells were co-cultured with irradiated J2NIH-3T3 fibroblasts (feeder cells) in DMEM complete medium with additives 10% FBS, penicillin, streptomycin, glutamine, F12 nutrient mixture (all 1X), 25. mu.g/m L hydrocortisone, 125ng/m L EGF, 5. mu.g/m L insulin, 250. mu.g/m L mycotoxin, 10. mu.g/m L gentamycin, 10. mu.g/m L statin, 0.1 nmol/L toxin and 10. mu.mol/L Rho kinase inhibitor Y27632 (Enzo). The culture medium was inoculated once every 3 days, and cell culture was passaged with trypsin for the following experiments L CR 5C.

FIG. 11 shows the CR L C cells obtained by reprogramming using the above conditions, wherein, after in vitro continuous culture for 3 months and more than 20 times, CR L C cells also have strong proliferation capacity (FIG. 11A). the results of cellular immunofluorescent labeling show that, after ten passages, the cells cultured in vitro are highly purified, the biomarkers CK7, CEA and TTF-1 of epithelial-derived tissues and non-small cell lung cancer are positive (FIG. 11B). immunohistochemistry and immunoblotting of paraffin-embedded tissues also confirm the expression of these markers (FIGS. 11C and 11D). DNA fingerprinting data show that, in FIG. 11E, there is no cell line contamination in early, middle and late passage CR L C cultures, and CR L C cells are successfully constructed and can be used in the following experiments.

Immunocytochemistry, namely fixing cells by using 4% paraformaldehyde buffer solution for 15 minutes, adding 1% Triton X-100, incubating for 10min at room temperature, washing for 5min × 3 times by PBS, sealing for 30min at room temperature by 3% BSA, dropwise adding anti-CEA, CK7 and TTF-1 on a cell slide, flatly placing the cell slide in a wet box, incubating overnight at 4 ℃, washing for 3 times by PBS every 5min every other day, dropwise adding a secondary antibody to cover the slide, incubating for 50min at room temperature, washing for 3 times by PBS, washing for 5min every time, adding DAB developing solution into the slide, controlling the developing time under a microscope, wherein the positive is brown yellow, the developing is stopped by washing the slide with tap water, performing hematoxylin-situ repolarization for about 3min, washing with tap water, differentiating for seconds by 1% hydrochloric acid alcohol, washing with tap water, returning ammonia water to blue, washing with running water, photographing with alcohol, sealing with neutral gum, and finally observing the dyeing condition of the cells under.

Immunofluorescence labeling: cells were fixed with 4% paraformaldehyde and non-specific antibody binding was blocked by incubation with 5% normal goat serum in PBS. The cells were then conjugated with primary antibody, conjugated with red fluorescence

594 conjugated secondary antibodies (Abcam, MA, USA) were incubated together and finally the cells were stained with DAPI for 10min at room temperature. Observations were made using a Zeiss fluorescence microscope equipped with a fluorescein filter set at 350nm (blue) and 488nm (red).

STR analysis: short Tandem Repeat (STR) analysis (i.e., DNA fingerprinting) A commercially available kit (Cell IDSystems; Promega Corporation, Madison, Wis.) was used. The system can carry out co-amplification and trichromatic detection on 10 loci (9 STR loci and Y chromosome specific Amelogenin). Wherein, STR mark: CSF1PO, D13S317, D16S539, D5S818, D7S820, THO1, TPOX, vWA. PCR amplification is according to the manufacturer's recommended protocol. The amplified fragments were detected using ABI3100 gene analyzer (Applied Biosystems). Data analysis and allele size determination were performed using GeneMapper software (Applied Biosystems).

Apoptosis analysis by flow cytometry CR L Cs were seeded into six well plates and allowed to attach for 24 hours, treated with PRIS for 48 hours, and apoptosis assessed by the Annexin V-FITC/PI apoptosis detection kit following the manufacturer's instructions (Miltenyi, Bergisch Gladbach, Germany.) briefly, cells were trypsinized, washed with PBS, resuspended in binding buffer, and incubated with Annexin V-FITC and PI for 10 minutes at room temperature in the dark flow cytometry analysis used the L SRII system (BD L SRII)^TMGermany) and FlowJo software (Tree Star, usa).

PRIS treatment PRIS was purchased from Sigma, dissolved in sterile dimethyl sulfoxide (DMSO) and stored at-20 ℃ in a solution of 50 mmol/L.

Cell Proliferation Assay after treating CR L C with PRIS for an appropriate time, the number of cells was determined using the MTS Assay (CellTiter 96Aqueous One Solution Cell Proliferation Assay, Promega, Madison, Wis., USA) according to the manufacturer's instructions, the absorbance was measured at 490nm using a spectrophotometer, all experiments were repeated, and each concentration was read three times.

Cell migration experiments invasion and migration experiments were performed using the transwell cell method, briefly, top inoculation 5 × 10⁴Cells, supplemented with serum-free medium, were packed in 600 μ L DMEM with 10% FBS in the lower chamber, and after 48 hours, the lower chamber cells were fixed, stained in 0.1% crystal violet solution for 30 minutes, and then counted.

In vitro angiogenesis assay: in vitro angiogenesis Using the BD Bio-Coat angiogenesis System according to the manufacturer's instructionsTo determine, briefly, 2 × 10⁴Primary HUVECs (C-12200, Promocell, Germany) of individual cells/m L were seeded into matrigel-precoated well plates with CR L C and medium with or without PRIS, and after 18 hours the formation of vascular structures in vitro was assessed and photographically imaged using a fluorescence microscope (Zeiss Axio Observer Z1, Germany).

siRNA transfection cells were transfected with 5 × 10⁵After overnight culture, CR L c was transfected with L ipofectamine RNAi Max reagent (Invitrogen, Carlsbad, CA, USA) according to targeting EphB4 or interference control siRNA (scr siRNA) — protein levels were checked by quantitative real-time RT-PCR (qRT-PCR) and immunoblotting, all experiments performed 48 hours after transfection.

RNA extraction and qRT-PCR: total RNA was isolated by using the RNeasy Mini Kit (QIAGEN, Hilden, Germany). Total RNA (300ng) from each sample was reverse transcribed using a cDNA reverse transcription kit (Applied Biosystems, Foster City, Calif., USA) according to the manufacturer's protocol. SYBR Premix Ex Taq was used^TMII kit (applied biosystems, Foster City, Calif., USA) qRT-PCR amplification was performed and CFX96 was used^TMThe real-time PCR detection system was used for analysis, data were normalized with GAPDH as a reference under a 25. mu. L system (95 ℃, 30 s; 40 cycles: 95 ℃ for 5s, 60 ℃ for 30s), and the relative expression was calculated by the 2-. DELTA.CT method.

Western blot analysis samples containing equal amounts of protein (30. mu.g) were separated electrophoretically on polyacrylamide SDS gels and transferred by electroblotting onto polyvinylidene fluoride (PVDF, Amersham, UK) membranes, which were then saturated with 5% (v/v) FBS and probed with primary antibodies overnight at 4 ℃ the membranes were then washed thoroughly and probed with horseradish peroxidase conjugated secondary antibodies (Cell Signaling Technologies, Beverly, MA, USA) followed by enhanced chemiluminescence (GE healthcare, Buckinghamshire, UK) detection, membrane stripping and re-probing of total protein or GAPDH, quantitative intensity value bands for the corresponding protein loading controls by Quantity One 4.6.2 software (Bio-Rad L assays, Hercules, CA, USA), all immunoblots being representative of at least three independent experiments.

Caspase activity assay CR L C with or without PRIS was harvested and suspended in lysis buffer and incubated for 1h on ice, cell lysates were centrifuged at 4 deg.C (12,000 × g, 30 min), after centrifugation, supernatants were collected and protein concentrations were measured immediately using BCA assay kit for caspase activity assay, cell lysates were placed in 96-well plates and incubated with specific caspase substrate, Ac-L EHD-AMC for caspase 9 substrate, Ac-DEVD-AMC for caspase-4 substrate, Ac-L EVD-AFC for 1 hour at 37 deg.C, and fluorescence activity was measured at excitation/emission 380/440 nm.

Statistical analysis: unless otherwise stated, data are presented as mean ± standard deviation. Inter-group statistics were analyzed by one-way anova, followed by Student's t-test using the SPSS 16.0 software package. Compared to the indicated groups, the significance level was indicated by P <0.05, P < 0.01.

The above description is only exemplary of the present invention, and the protection scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the protection scope of the present invention.

Claims

2. Use according to claim 1, characterized in that: the EphB4/Ephrin-B2 signaling pathway inhibitors include EphB4 inhibitors and/or Ephrin-B2 inhibitors.

3. Use according to claim 1, characterized in that: EphB4/Ephrin-B2 signaling pathway inhibitors are useful for treating diseases in which the EphB4/Ephrin-B2 signaling pathway is abnormally activated.

4. Use according to claim 3, characterized in that: the diseases with abnormal activation of EphB4/Ephrin-B2 signal pathway comprise tumor diseases.

5. Use according to claim 4, characterized in that: the neoplastic disease includes head and neck cancer, ovarian cancer, lung cancer, esophageal cancer, or colorectal cancer.

6. Use according to claim 5, characterized in that: the lung cancer comprises non-small cell lung cancer and/or small cell lung cancer.

7. Use according to claim 4, characterized in that: the EphB4/Ephrin-B2 signal channel inhibitor can inhibit tumor cell proliferation, inhibit tumor cell migration, inhibit formation of tumor vascular structure and promote tumor cell apoptosis by inhibiting EphB4/Ephrin-B2 signal channel.

8. Use according to claim 1, characterized in that: the platypodium and/or the derivative thereof can reduce the expression level of EphB4 by at least 10%.

9. Use according to claim 1, characterized in that: the platypodium and/or the derivative thereof can reduce the expression level of Ephrin-B2 by at least 10%.

10. Use according to claim 1, characterized in that: the derivative of the plastic wisteria comprises pharmaceutically acceptable salt of the plastic wisteria, pharmaceutically acceptable ester of the plastic wisteria, structural analogues of the plastic wisteria, or a modifier of the plastic wisteria.