Anti-tumor composition and application thereof

Abstract

The invention provides an anti-tumor composition and application thereof, and belongs to the technical field of biological medicine. The invention provides an anti-tumor composition. The anti-tumor composition comprises a first component and a second component, the first component is prepared from 2-hydroxycinnamic acid; the second component comprises ferulic acid and / or cinnamic acid; the molar ratio of the first component to the second component is (1-2): (1-3). In the composition disclosed by the invention, the 2-hydroxycinnamic acid and the cinnamic acid can target EGFR (Epidermal Growth Factor Receptor), and at an EGFR binding site, the 2-hydroxycinnamic acid and the cinnamic acid enhance the binding stability through hydrophobic interaction; 2-hydroxycinnamic acid and ferulic acid can target PCNA, and at a PCNA binding site, 2-hydroxycinnamic acid and ferulic acid achieve a synergistic effect through pi-pi stacking interaction. The anti-tumor composition provided by the invention can be used for targeted therapy of tumors with EGFRY845 phosphorylation and / or PCNA high expression characteristics.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to an anti-tumor composition and application thereof. Background Art

[0002] Malignant tumors pose a major global public health challenge. Traditional drug treatment strategies often exhibit limitations in the face of tumor heterogeneity and complexity. Effective targeted therapeutic strategies are currently lacking for tumors characterized by EGFRY845 phosphorylation and / or elevated PCNA expression. Summary of the Invention

[0003] The object of the present invention is to provide an anti-tumor composition and its application, which can target and treat tumors characterized by EGFRY845 phosphorylation and / or high PCNA expression.

[0004] The present invention provides an antitumor composition comprising a first component and a second component;

[0005] The first component includes 2-hydroxycinnamic acid;

[0006] The second component includes ferulic acid and / or cinnamic acid;

[0007] The molar ratio of the first component to the second component is (1-2): (1-3).

[0008] Preferably, the anti-tumor composition comprises the following components: ferulic acid, 2-hydroxycinnamic acid and cinnamic acid.

[0009] Preferably, the molar ratio of ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is (1-2):(1-2):1.

[0010] Preferably, the molar ratio of ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is 1:1:1.

[0011] Preferably, the ferulic acid is replaced by methyl ferulate or caffeic acid.

[0012] The present invention also provides an anti-tumor drug delivery system, the active ingredient of which includes the anti-tumor composition described in the above scheme.

[0013] Preferably, the anti-tumor drug delivery system comprises a liposome delivery system;

[0014] The liposome delivery system comprises the following raw materials: lecithin, cholesterol, the antitumor composition according to any one of claims 1 to 5, and a solvent;

[0015] The mass ratio of the lecithin, cholesterol and anti-tumor composition is 5:1:1.

[0016] The application further provides application of the anti-tumor composition or the anti-tumor drug delivery system in the preparation of an anti-tumor drug.

[0017] Preferably, the tumor comprises a tumor with EGFRY845 phosphorylation and / or PCNA high expression.

[0018] Preferably, the tumor comprises at least one of gastric cancer, lung cancer and breast cancer.

[0019] The application provides an anti-tumor composition comprising a first component and a second component; the first component comprises 2-hydroxycinnamic acid; the second component comprises ferulic acid and / or cinnamic acid; and the molar ratio of the first component to the second component is (1-2):(1-3). EGFRY845 site phosphorylation is related to disease progression and poor prognosis in various tumors such as gastric cancer, lung cancer and breast cancer, and is also an indicator of drug resistance. PCNA, as a marker of proliferating cells, is closely related to the over-proliferation of tumor cells. In the composition of the application, 2-hydroxycinnamic acid and cinnamic acid can target EGFR, and at the EGFR binding site, 2-hydroxycinnamic acid and cinnamic acid enhance the binding stability through hydrophobic interaction; and 2-hydroxycinnamic acid and ferulic acid can target PCNA, and at the PCNA binding site, 2-hydroxycinnamic acid and ferulic acid achieve synergistic effect through π-π stacking interaction. The anti-tumor composition of the application can target the treatment of tumors with EGFRY845 phosphorylation and / or PCNA high expression characteristics. BRIEF DESCRIPTION OF DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings in the following description are only some embodiments of the application, and for those skilled in the art, other drawings can also be obtained from these drawings without creative labor.

[0021] Figure 1 Dose-dependent inhibition curve of three compounds and the combination group on HGC27 cells;

[0022] Figure 2 Dose-dependent inhibition curve of three compounds and the combination group on AGS cells;

[0023] Figure 3 Combination index (CI) value of DCF in HGC27 and AGS cells;

[0024] Figure 4 CCK8 method for detecting the experimental results of the cytotoxicity of the related drugs;

[0025] Figure 5 This is the experimental result diagram of the CCK8 method for detecting the cytotoxicity of related drugs;

[0026] Figure 6 This is the experimental result diagram of the CCK8 method for detecting the cytotoxicity of related drugs;

[0027] Figure 7 This is the molecular docking model of DCF compound and EGFR;

[0028] Figure 8 This is the molecular docking model of DCF compound and PCNA;

[0029] Figure 9 is the inhibitory effect of DCF on EGFRY845 phosphorylation;

[0030] Figure 10 is the interference effect of DCF on PCNA-CDK2 interaction;

[0031] Figure 11 TEM image of DCF liposome delivery system;

[0032] Figure 12 is the particle size distribution of DCF liposome delivery system;

[0033] Figure 13 The antitumor effect of DCF liposome delivery system in gastric cancer PDX model;

[0034] Figure 14 To immunohistochemically analyze the effects of DCF liposome delivery system on EGFRY845 phosphorylation and PCNA expression in tumor tissues;

[0035] Figure 15 This is the experimental result of detecting the toxic effects of 2-hydroxycinnamic acid and cinnamic acid combination on HGC27 cells with high EGFRY845 phosphorylation level by CCK8 method;

[0036] Figure 16 This is the experimental result of detecting the toxic effect of the combination of 2-hydroxycinnamic acid and ferulic acid on AGS cells with high PCNA levels using the CCK8 method;

[0037] Figure 17 The figure shows the experimental results of the toxic effects of 2-hydroxycinnamic acid, cinnamic acid and ferulic acid (molar ratio of 1:2:1) on HGC27 detected by CCK8 method;

[0038] Figure 18 The figure shows the experimental results of the toxic effects of 2-hydroxycinnamic acid, cinnamic acid and ferulic acid (molar ratio of 2:1:1) on HGC27 detected by CCK8 method;

[0039] Figure 19 Figure for CCK8 method to detect the toxicity of 2-hydroxycinnamic acid, cinnamic acid and ferulic acid methyl ester (molar ratio of 1:1:1) on HGC27;

[0040] Figure 20 Figure for CCK8 method to detect the toxicity of 2-hydroxycinnamic acid, cinnamic acid and caffeic acid (molar ratio of 1:1:1) on HGC27;

[0041] Figure 21 Figure for CCK8 method to detect the cytotoxicity of DCF combined with cisplatin on HGC27;

[0042] Figure 22 Figure for CCK8 method to detect the cytotoxicity of DCF combined with PD-L1 immune checkpoint inhibitor MDX-1105 on HGC27;

[0043] Figure 23 Figure for CCK8 method to detect the cytotoxicity of DCF on EGFR-TKI resistant cell lines H460 and A549. DETAILED DESCRIPTION

[0044] The present application provides an anti-tumor composition (DCF), comprising a first component and a second component; the first component comprises 2-hydroxycinnamic acid; the second component comprises ferulic acid and / or cinnamic acid; the molar ratio of the first component and the second component is (1-2):(1-3).

[0045] In the present application, the chemical structural formula of ferulic acid is shown as formula I:

[0046]

[0047] In the present application, the chemical structural formula of 2-hydroxycinnamic acid is shown as formula II:

[0048]

[0049] In the present application, the chemical structural formula of cinnamic acid is shown as formula III:

[0050]

[0051] In the present invention, at the molecular level, 2-hydroxycinnamic acid, ferulic acid, and cinnamic acid are all phenolic acid compounds with similar core structures, but their different substituents give them complementary target affinities. Ferulic acid contains methoxy and hydroxyl substitutions, enhancing its binding to PCNA; 2-hydroxycinnamic acid, with a hydroxyl group at the 2-position, can bind simultaneously to both EGFR and PCNA. Cinnamic acid has the simplest structure and primarily binds to EGFR. This structural complementarity enables 2-hydroxycinnamic acid, ferulic acid, and cinnamic acid to cover different binding sites, enhancing their overall efficacy. Secondly, at the target level, 2-hydroxycinnamic acid, ferulic acid, and cinnamic acid form a synergistic "division of labor and cooperation" model: 2-hydroxycinnamic acid and cinnamic acid primarily target EGFR, while 2-hydroxycinnamic acid and ferulic acid primarily target PCNA. In particular, 2-hydroxycinnamic acid simultaneously participates in the action of both targets, acting as a "bridge." This target complementarity enables the anti-tumor composition to effectively inhibit both key targets. Furthermore, at the pathway level, DCF simultaneously targets both the apoptosis pathway (EGFR-Bcl2 / Bax) and the cell cycle pathway (PCNA-CDK2), creating a "two-pronged" synergistic effect. These two pathways are independent yet mutually reinforcing. Even if tumor cells develop resistance to one mechanism, the other remains effective, significantly reducing the risk of resistance.

[0052] Phosphorylation of the EGFRY845 site is associated with disease progression and poor prognosis in various tumors such as gastric cancer, lung cancer, and breast cancer, and is also an indicator of drug resistance. In the present invention, the anti-tumor composition can inhibit EGFRY845 phosphorylation and trigger tumor cell apoptosis. Specifically, in terms of EGFR targeting, unlike traditional EGFR inhibitors, 2-hydroxycinnamic acid and cinnamic acid can directly bind to the pocket where the Y845 site of EGFR is located, rather than the ATP binding site or tyrosine kinase domain, by forming hydrophobic interactions with Leu694, Val702, Leu694 and Leu768, stably binding to the pocket near Y845, preventing Src kinase-mediated phosphorylation of the Y845 site. This unique binding mode enables the anti-tumor composition to specifically inhibit EGFRY845 phosphorylation without affecting the total expression of EGFR and phosphorylation of other sites, thereby effectively reducing the level of EGFRY845 phosphorylation, avoiding some adverse reactions of traditional EGFR inhibitors, and at the same time potentially overcoming the drug resistance of some EGFR targeted drugs (EGFR targeted therapy, hormone therapy and chemotherapy). Inhibition of Y845 phosphorylation is the key mechanism by which anti-tumor compositions induce tumor cell apoptosis.

[0053] PCNA is a marker for proliferating cells, and its high expression is closely associated with the excessive proliferation of tumor cells. In the present invention, the anti-tumor composition can interfere with the PCNA-CDK2 interaction, causing S-phase arrest. Specifically, in terms of PCNA targeting, the anti-tumor composition can directly bind to the PCNA protein, and its binding site significantly overlaps with the PCNA-CDK2 interaction interface. 2-Hydroxycinnamic acid forms a hydrogen bond with PCNA's Arg149 through its carboxyl group, while ferulic acid forms a stable bond by forming hydrophobic interactions with Met40 and Val45, effectively weakening PCNA-CDK2 binding. The Arg149 residue is a key site for 2-Hydroxycinnamic acid and ferulic acid to exert their effects. The anti-tumor composition can bind to a specific site on PCNA, interfering with the PCNA-CDK2 interaction. This interference differs from traditional PCNA inhibitors (such as PCNA-I1), which primarily bind to the PCNA trimer interface. The unique mode of action of the anti-tumor composition of the present invention specifically blocks CDK2-dependent S-phase progression, leading to S-phase arrest and significant accumulation of tumor cells in the S-phase without affecting other PCNA functions, thereby effectively inhibiting tumor cell DNA synthesis. The anti-tumor composition of the present invention effectively reduces the binding of PCNA to CDK2 without affecting the interaction between PCNA and p21.

[0054] As an embodiment, the purity of the ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is respectively ≥98%.

[0055] In the present invention, ferulic acid, 2-hydroxycinnamic acid and cinnamic acid are widely present in a variety of foods and medicinal plants and have good safety.

[0056] As an embodiment, the anti-tumor composition includes the following components: ferulic acid, 2-hydroxycinnamic acid and cinnamic acid.

[0057] In the present invention, 2-hydroxycinnamic acid and cinnamic acid can target EGFR, while 2-hydroxycinnamic acid and ferulic acid can target PCNA, forming complementary interactions at the target sites. Furthermore, at the EGFR binding site, 2-hydroxycinnamic acid and cinnamic acid enhance binding stability through hydrophobic interactions; at the PCNA binding site, 2-hydroxycinnamic acid and ferulic acid achieve a synergistic effect through π-π stacking interactions. Furthermore, as another embodiment, the anti-tumor composition comprises the following components: ferulic acid, 2-hydroxycinnamic acid, and cinnamic acid.

[0058] In the present application, the anti-tumor composition can both inhibit EGFR Y845 phosphorylation to induce tumor cell apoptosis and interfere with PCNA-CDK2 interaction to cause S phase arrest. This dual molecular mechanism produces a series of cascade reactions: EGFR Y845 phosphorylation inhibition leads to down-regulation of Bcl-2 and up-regulation of Bax, triggering mitochondrial-mediated apoptosis; PCNA-CDK2 interaction interference leads to S phase arrest, blocking DNA replication and proliferation of tumor cells. The two mechanisms synergize with each other, and ferulic acid, 2-hydroxycinnamic acid and cinnamic acid simultaneously inhibit the EGFR-mediated Bcl-2 / Bax apoptosis pathway and the PCNA-CDK2-mediated S phase arrest pathway, thereby achieving efficient inhibition of EGFR Y845 phosphorylation and PCNA high-expression tumors and exerting synergistic anti-tumor effects.

[0059] In the present application, ferulic acid, 2-hydroxycinnamic acid and cinnamic acid can exert synergistic anti-tumor effects, with a CI value less than 0.8.

[0060] As an embodiment, the molar ratio of ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is (1-2) : (1-2) : 1.

[0061] As an embodiment, the molar ratio of ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is 1:1:1, which has the best synergistic anti-tumor effect.

[0062] As an embodiment, the ferulic acid is replaced by methyl ferulate or caffeic acid.

[0063] As an embodiment, the preparation method of the anti-tumor composition comprises the following steps: dissolving the first component and the second component in a solvent respectively and then mixing them. As an embodiment, the solvent comprises DMSO or anhydrous ethanol.

[0064] As an embodiment, the mixture can be used directly or further processed into a specific dosage form after the mixing.

[0065] The present application also provides an anti-tumor drug delivery system, and the active ingredient comprises the anti-tumor composition described in the above scheme.

[0066] As an embodiment, the anti-tumor drug delivery system comprises a liposome delivery system, a solid lipid nanoparticle (SLN), a polymer nanoparticle, a targeted delivery system or a stimulus-responsive delivery system.

[0067] As an embodiment, the SLN has better stability and controllable release characteristics, is suitable for oral administration and is suitable for clinical application of anti-tumor drugs.

[0068] As an embodiment, polymer nanoparticles are nanoparticle systems based on biocompatible polymers such as PLGA and PCL, which can achieve slow release of anti-tumor drugs and prolong the duration of action.

[0069] As an embodiment, the targeted delivery system is to modify specific targeting ligands on the surface of liposomes to achieve active targeting of tumor tissues and further increase the concentration of DCF at the tumor site; the specific targeting ligands include at least one of folic acid, transferrin and specific antibodies.

[0070] As an embodiment, the stimuli-responsive delivery system is an intelligent delivery system that responds to pH, temperature or enzymes, and can achieve the specific release of DCF in the tumor microenvironment and reduce drug exposure in normal tissues.

[0071] As an embodiment, the liposome delivery system includes the following raw materials: lecithin, cholesterol, the anti-tumor composition described in the above scheme and a solvent; the mass ratio of the lecithin, cholesterol and anti-tumor composition is 5:1:1; the solvent includes chloroform; the lecithin and cholesterol are both FDA-approved safe ingredients, which are conducive to subsequent clinical translation research.

[0072] The liposome delivery system of the present invention overcomes the shortcomings of poor water solubility and low bioavailability of phenolic acid compounds, and can significantly improve the bioavailability and antitumor effect of the antitumor composition.

[0073] From the perspective of physicochemical properties, the liposome delivery system of the present invention is spherical, with a smooth surface, a particle size of 80 to 120 nm, and an average particle size of 102.60 ± 1.55 nm, which is within the ideal nano-delivery range (50 to 200 nm). This particle size is small enough to avoid rapid clearance by the reticuloendothelial system, but large enough to passively target and enrich at the tumor site through the enhanced permeability and retention effect (EPR). The liposome delivery system of the present invention is stored at 4°C and 25°C for 30 days, and the particle size changes by less than 5%, and the drug content decreases by less than 10%. The liposome delivery system also exhibits good stability in different pH (5.0 to 8.0) and serum environments. The zeta potential (-35.37 mV) of the liposome delivery system of the present invention is in the stable range, the polydispersity coefficient (PDI) is 0.185 ± 0.004, and the encapsulation efficiency of ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is more than 80%, showing good stability.

[0074] From the perspective of pharmacokinetic parameters, the anti-tumor composition is encapsulated by liposomes, which prolongs the half-life of each component of the anti-tumor composition, prolongs the average residence time, increases the AUC, and improves the bioavailability of the anti-tumor composition.

[0075] From the perspective of cellular level effects, the liposome delivery system of the present invention can significantly improve the cellular uptake efficiency of the anti-tumor composition.

[0076] In terms of in vivo anti-tumor effects, the tumor inhibition rate of the liposome delivery system of the present invention is higher than that of the free anti-tumor composition. Moreover, it maintains a high therapeutic effect while reducing the dosage, which is beneficial for reducing potential toxic side effects.

[0077] In one embodiment, the method for preparing the liposome delivery system comprises the following steps: preparing the liposome delivery system using a thin film hydration method: dissolving lecithin, cholesterol, and an anti-tumor composition in a solvent, rotary evaporating the solution to form a thin film, drying the solution with nitrogen gas, hydrating the solution with PBS, and ultrasonically treating the solution before filtering the solution through a 0.22 μm filter membrane to obtain the liposome delivery system. In one embodiment, the rotary evaporation is performed using a rotary evaporator.

[0078] The present invention also provides the use of the antitumor composition or the antitumor drug delivery system described in the above scheme in the preparation of antitumor drugs.

[0079] As an embodiment, the anti-tumor composition or the anti-tumor drug delivery system is the only active ingredient of the anti-tumor drug.

[0080] As another embodiment, in addition to the anti-tumor composition or the anti-tumor drug delivery system, the anti-tumor drug further comprises other anti-tumor components to achieve a synergistic effect of combined therapy; the other anti-tumor components comprise chemotherapy drugs and / or other targeted drugs; the chemotherapy drugs comprise cisplatin and / or paclitaxel; the other targeted drugs comprise PD-L1 immune checkpoint inhibitors, and further comprise the PD-L1 immune checkpoint inhibitor MDX-1105;

[0081] In one embodiment, the tumor includes a tumor in which EGFRY845 is phosphorylated and / or PCNA is highly expressed.

[0082] As an embodiment, the anti-tumor composition includes three components: ferulic acid, 2-hydroxycinnamic acid and cinnamic acid. By inhibiting EGFRY845 phosphorylation and interfering with PCNA-CDK2 interaction, it can be used to treat tumors with EGFRY845 phosphorylation and high PCNA expression.

[0083] EGFRY845 phosphorylation and PCNA overexpression are common features of various tumors, not only in gastric cancer, but also in lung cancer, breast cancer, colorectal cancer and other malignant tumors. The specific effect of the anti-tumor composition on these two markers makes it potentially suitable for the treatment of various tumor types. As an embodiment, the tumor includes at least one of gastric cancer, lung cancer, breast cancer, oral squamous cell carcinoma, liver cancer and colorectal cancer; the gastric cancer includes gastric cancer cell lines HGC27 or AGS; the colorectal cancer includes colon cancer cell line HT-29; the lung cancer includes non-small cell lung cancer; the non-small cell lung cancer includes cell line A549.

[0084] As an embodiment, the anti-tumor drug still has good inhibitory effect on EGFR-TKI resistant cell lines, indicating its potential resistance overcoming effect; the EGFR-TKI resistant cell lines include H460 and / or A549.

[0085] The anti-tumor composition of the present application has CI values of 0.703 and 0.528 for gastric cancer cell lines HGC27 and AGS, respectively, both of which are less than 1, and has significant synergistic effect. This synergistic effect enables the anti-tumor composition to achieve significant anti-tumor effect at a lower concentration, reducing potential toxic side effects.

[0086] As an embodiment, the drug further includes a pharmaceutically acceptable salt and one or more pharmaceutically acceptable carriers, excipients or diluents; the drug can be prepared into solid oral preparations, liquid oral preparations or injections; the injections include intravenous injection forms; the drug can also be administered by parenteral forms, preferably by injection; the solid and liquid oral preparations include tablets, capsules, syrup, granules or oral solutions.

[0087] As an embodiment, the preparation of the drug includes the following steps: using standard and conventional techniques to combine the anti-tumor composition of the present application with pharmaceutically acceptable solid or liquid carriers, and optionally with pharmaceutically acceptable adjuvants and excipients to prepare microparticles or microspheres. Solid dosage forms include tablets, capsules, sustained-release tablets, sustained-release pellets and the like. Solid carriers can be at least one substance that can act as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, disintegrant and coating agent. Inert solid carriers include at least one of magnesium phosphate, magnesium stearate, talc, lactose, pectin, propylene glycol, polysorbate 80, dextrin, starch, gelatin, cellulose substances such as methyl cellulose, microcrystalline cellulose, low-melting paraffin, polyethylene glycol, mannitol and cocoa butter. Liquid dosage forms include solvents or suspensions or injections.

[0088] EGFRY845 phosphorylation and high PCNA expression level are positively correlated with the therapeutic effect of the anti-tumor drugs in the above regimen.

[0089] The application also provides a method for personalized treatment of tumors, comprising the following steps: predicting the sensitivity of a patient to the treatment of the anti-tumor composition or the anti-tumor drug delivery system in the above regimen by detecting the EGFRY845 phosphorylation and PCNA expression level in tumor tissue, thereby realizing personalized treatment.

[0090] The method for personalized treatment of tumors of the application is a precise treatment strategy.

[0091] The method for personalized treatment of tumors of the application can be used in combination with existing treatment methods, as a component of combination therapy, and provides a new strategy for comprehensive treatment of tumors; the existing treatment methods include at least one of surgery, radiotherapy, chemotherapy and immunotherapy.

[0092] Explanation of terms:

[0093] Ferulic acid: a naturally occurring phenolic compound, chemical name 4-hydroxy-3-methoxycinnamic acid, molecular formula C 10 H 10 O4, has multiple biological activities, including antioxidant, anti-inflammatory and anti-tumor effects.

[0094] 2-Hydroxycinnamic acid: also known as o-hydroxycinnamic acid, is a hydroxyl derivative of cinnamic acid, molecular formula C9H8O3, has antioxidant and anti-tumor activity.

[0095] Cinnamic acid: an aromatic unsaturated carboxylic acid, molecular formula C9H8O2, widely exists in various plants, has antibacterial, antioxidant and anti-tumor activity.

[0096] EGFRY845 phosphorylation: phosphorylation modification of the 845th tyrosine residue of the epidermal growth factor receptor (EGFR), is an important marker of EGFR activation, and is closely related to the malignant progression and drug resistance of various tumors.

[0097] PCNA (proliferating cell nuclear antigen): a protein that plays a key role in DNA replication and repair, is an important marker of cell proliferation, and is highly expressed in various tumors.

[0098] CI value (combination index): a parameter for evaluating the combined effect of drugs, CI <1 indicates synergistic effect, CI =1 indicates additive effect, and CI >1 indicates antagonistic effect.

[0099] To further illustrate the present invention, an anti-tumor composition and its application provided by the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0100] Example 1

[0101] Raw materials: 2-hydroxycinnamic acid, cinnamic acid and ferulic acid, with a molar ratio of 1:1:1.

[0102] 2-Hydroxycinnamic acid, cinnamic acid and ferulic acid are dissolved in anhydrous ethanol and then mixed to obtain an anti-tumor composition DCF.

[0103] Test Example 1

[0104] Evaluation of the in vitro antitumor activity of DCF in Example 1

[0105] 1. Cell survival and proliferation inhibition

[0106] CCK-8 pharmacodynamics validation (including gastric cancer cells and other tumor cells)

[0107] Here are the steps:

[0108] (a) 96-well plate plating: Human gastric cancer cell lines HGC-27 and AGS, colon cancer cell line HT-29, and lung cancer cell line A549 were trypsinized and resuspended from the cell culture dish. 10 μL of the cell suspension (1:1 dilution) was counted using a cell counting plate and seeded into 96-well plates at a density of 5000 cells / well. The plates were then placed in a cell culture incubator and allowed to adhere.

[0109] (b) Drug Addition: The next day, after the cells have adhered to the wall, replace each well with 200 μL of fresh serum-free medium. Adjust the final drug concentrations to 0, 0.1875, 0.375, 0.75, 1.5, 3, and 5 mM. Set up three replicate wells for each concentration group, and include a blank well (without cells) for background subtraction and zeroing. Incubate the cells in a 37°C, 5% CO2 incubator for 24 h.

[0110] (c) Detection of cell activity: Dilute the CCK-8 reagent with fresh serum-free medium at a volume ratio of 10:1 to prepare a working solution. After 24 hours of drug intervention, remove the supernatant from each well, add 100 μL of CCK-8 working solution to each well, and continue to incubate in a 37°C cell culture incubator for 1.5 hours before taking out. Set the wavelength parameter of the microplate reader to 450 nm, read the absorbance (OD) value, and use GraphPad Prism v6.0 software to draw a graph and calculate the half-maximal inhibitory concentration (IC) corresponding to 24 hours. 50 ).

[0111] CCK8 experiment evaluated the cytotoxicity of DCF on gastric cancer cells (HGC27 and AGS) ( Figure 1 and Figure 2 ), lung cancer cells (A549) and colon cancer cells (HT-29) ( Figure 4 ). The results showed that DCF exhibited dose-dependent growth inhibition on the four cell lines, and the three drugs showed good synergistic effect on human gastric cancer cells HGC27 and AGS cells in the concentration range of 0.1875-1.5 mM; when the concentration reached 5 mM, the synergistic effect of the three drugs was not good; and human colon cancer cells HT-29 and human lung cancer cell line A549 (both EGFRY845 and PCNA expression high level) in the concentration range of 0.1875-1.5 mM, the three drugs showed good synergistic effect, and showed stronger inhibition effect on EGFRY845 phosphorylation and PCNA high expression cell lines, thus showing that the drug combination has good specific therapeutic effect on EGFRY845 and PCNA high expression tumor ( Figure 5 ). In HGC27 and AGS cells, the 24h IC 50 values of DCF were 0.680±0.052 mM and 0.744±0.024 mM, respectively ( Figure 3 ). In addition, human gastric cancer cells HGC27 in the concentration range of 0.1875-1.5 mM, the three drugs showed good synergistic effect; while the effect of two drug combination (ferulic acid+cinnamic acid) was not as good as that of three drug combination, indicating that the effect of DCF three drug combination group was better than that of each single drug group and double drug group ( Figure 6 ).

[0112] 2. Molecular docking verification

[0113] The protein target and small molecule are docked by using Autodock software, and at least 50 docking results will be generated, so as to select the best phase of docking energy for structure extraction for subsequent molecular dynamics simulation research. Autodock is an open source small molecule simulation software, which is mainly used to perform molecular docking between ligand and protein. The basic process of docking is as follows: ① a larger box is formed around the amino acid residues of the receptor active site, and then different types of atoms are used as probes for scanning, so as to calculate the grid energy. This part of the task is completed by AutoGrid program; ② AutoDock program performs conformation search of ligand in Box range, and finally the ligand is scored according to different conformation, direction, position and energy, and the results are optimized and sorted.

[0114] Through molecular docking analysis, the results showed that the three kinds of compound components involved in DCF could bind to EGFR and PCNA protein targets to different degrees (​​​​​ Figure 7 and Figure 8 ).

[0115] 3. Molecular mechanism validation

[0116] Western blotting method for detecting expression levels of related proteins in gastric cancer cells

[0117] Blank control group and related drug concentration treatment group were set for two kinds of gastric cancer cells respectively, and then the cells were treated for 24 h. Then the total protein of the cells after drug treatment was extracted and labeled by BCA protein concentration, and the absolute protein content of each well was ensured to be consistent. After electrophoresis, the gel segment containing the target protein was cut according to the pre-stained protein marker. Then constant voltage was applied for membrane transfer, and after washing and blocking the membrane, the corresponding protein one-antibody was added, and incubated overnight at 4°C. The next day, the second antibody was incubated after washing the membrane, and then the ECL chemiluminescence method was used for detection. The total protein of the cells was detected by mouse anti-β-actin monoclonal antibody as an internal control.

[0118] The effects of DCF treatment on the expression and phosphorylation of key proteins were analyzed by Western blot. The results showed that DCF treatment significantly reduced the phosphorylation level of EGFR Y845, but did not affect the phosphorylation of EGFR total protein and other sites (Y1068, Y1173), confirming the specific inhibitory effect of DCF on Y845 site Figure 9 ).

[0119] Co-immunoprecipitation experiment confirmed that DCF treatment significantly weakened the interaction between PCNA and CDK2, but did not affect the binding of PCNA to other proteins (such as p21), confirming the mechanism of DCF specific interference with PCNA-CDK2 interaction Figure 10 ).

[0120] Example 2

[0121] 1. Preparation of DCF liposome delivery system

[0122] Lecithin, cholesterol and DCF prepared in Example 1 (based on the total mass of 2-hydroxycinnamic acid, cinnamic acid and ferulic acid) were dissolved in chloroform at a mass ratio of 5:1:1, evaporated to form a thin film on a rotary evaporator, and then hydrated with PBS after nitrogen blowing. After ultrasonic treatment, the DCF liposome delivery system was obtained by filtering through a 0.22 μm filter membrane.

[0123] 2. Physicochemical characterization of DCF liposome delivery system

[0124] The DCF liposome delivery system was characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS) Figure 11 ) TEM observation showed that the DCF liposome delivery system was spherical and smooth. DLS measurement showed that the average particle size of the DCF liposome delivery system was 102.60 ± 1.55 nm, which belonged to the ideal nano delivery range (50-200 nm), and this particle size was small enough to avoid being quickly removed by the reticuloendothelial system and large enough to be passively targeted and enriched at the tumor site by the enhanced permeability and retention effect (EPR). The polydispersity index (PDI) was 0.185 ± 0.004 Figure 12 ).

[0125] 3. Stability evaluation of the DCF liposome delivery system

[0126] The particle size change of the DCF liposome delivery system was less than 5% and the drug content decreased by less than 10% after storage at 4°C and 25°C for 30 days, indicating good stability. The DCF liposome delivery system also showed good stability in different pH (5.0-8.0) and serum environments.

[0127] 4. Pharmacokinetic parameters

[0128] Liposome encapsulation prolonged the half-life of each component of DCF by 1.5-2 times, prolonged the mean residence time by 1.8-2.3 times, and increased the AUC by 1.5-2 times. The improvement of these parameters directly reflects the significant improvement of the bioavailability of DCF liposome delivery system.

[0129] Test Example 2

[0130] Evaluation of the anti-tumor activity of the DCF liposome delivery system of Example 2 in vivo

[0131] 4-week-old male NCG mice were purchased from Beijing Aidmo Biotechnology Co., Ltd. All 24 mice were randomly divided into 4 groups, 6 mice in each group. When they reached 5 weeks of age, the patient tumor mass was subcutaneously inoculated into the right hip region of the 24 mice using a trocar. After a period of time when the tumor was visible to the naked eye (1 cm), the mice in the cocktail and cocktail liposomal groups were treated with intraperitoneal injection of drugs, and the drugs were administered once a day during the experiment; at the same time, the positive drug control group involved oxaliplatin (L-OHP, 4 mg / kg body weight) and 5-fluorouracil (5-FU, 20 mg / kg body weight) were administered to the mice intraperitoneally, and the drugs were administered once every three days during the experiment. On the 14th day, all the mice were sacrificed at the end of the experiment, and the tumor specimens were photographed and the tumor tissue specimens were fixed and subsequently embedded, sectioned and stained. No mouse died unexpectedly during the entire experiment.

[0132] Immunohistochemical experimental method

[0133] Place the cut paraffin sections in an oven at 60-65°C until the water evaporates and the paraffin melts completely. Soak the paraffin sections in xylene solution for 15 minutes, remove them, and repeat the above steps after replacing the xylene solution to dissolve the excess paraffin. After rehydration with graded alcohol, hematoxylin staining, and dehydration, the sections are stained in eosin reagent for 1 minute and then dehydrated again. Finally, use a pipette to draw an appropriate amount of neutral gum droplet on the tissue, cover it with a coverslip, and gently tap with tweezers to remove any bubbles. Once the gum has dried, the sections can be observed immediately or stored long-term.

[0134] 1. In vivo antitumor activity

[0135] The antitumor activity of DCF was evaluated in a gastric cancer PDX mouse model based on EGFRY845 phosphorylation and high PCNA expression. When tumor volume reached approximately 100 mm3, mice were randomly divided into six groups (n=8): a control group, a low-dose free DCF group (10 mg / kg), a high-dose free DCF group (20 mg / kg), a low-dose DCF liposome delivery system group (10 mg / kg), a high-dose DCF liposome delivery system group (20 mg / kg), and a positive control group (oxaliplatin, 5 mg / kg). Dosing was performed three times weekly for three consecutive weeks.

[0136] The results showed that the high-dose group of the DCF liposome delivery system showed the strongest anti-tumor effect, with a tumor inhibition rate of 75.3±5.1%, which was significantly higher than the high-dose group of free DCF (52.8±4.7%) and the positive control group (61.5±5.4%). The anti-tumor effect of the low-dose group of the DCF liposome delivery system (49.6±4.2%) was comparable to that of the high-dose group of free DCF, indicating that liposome delivery can significantly enhance the in vivo anti-tumor effect of DCF. More importantly, the effect of the low-dose group of the DCF liposome delivery system (10 mg / kg) was comparable to that of the high-dose group of free DCF (20 mg / kg), indicating that liposome delivery can maintain a higher therapeutic effect while reducing the dose, which is beneficial to reduce potential toxic side effects ( Figure 13 ).

[0137] 2. In vivo mechanism verification

[0138] Immunohistochemical analysis showed that treatment with the DCF liposome delivery system significantly reduced the expression of Ki-67, phosphorylated EGFRY845, and PCNA in tumor tissues, confirming that the mechanism of action of DCF in vivo was consistent with the results of in vitro studies ( Figure 14 ).

[0139] Example 3

[0140] Raw materials: 2-hydroxycinnamic acid and cinnamic acid, molar ratio is 1:1.

[0141] 2-Hydroxycinnamic acid and cinnamic acid are dissolved in anhydrous ethanol and then mixed to obtain an anti-tumor composition DCF, which is used to treat tumors in which EGFRY845 phosphorylation is the main driving factor.

[0142] Test Example 3

[0143] CCK-8 pharmacodynamics verification of the anti-tumor effect of the anti-tumor composition of Example 3 (on HGC27 cells with high EGFRY845 phosphorylation levels)

[0144] Here are the steps:

[0145] (a) 96-well plate plating: After trypsinizing the cells from the cell culture dish and resuspending them, 10 μL of the cell suspension (1:1 dilution) was counted using a cell counting plate and seeded into a 96-well plate at a density of 5000 cells / well. The plate was then placed in a cell culture incubator and allowed to adhere.

[0146] (b) Drug Addition: The next day, after the cells have adhered to the wall, replace each well with 200 μL of fresh serum-free medium. Adjust the final drug concentration to 0, 1.5, and 3 mM. Set up three replicate wells for each concentration group, and include a blank well (without cells) for background subtraction and zeroing. Incubate the cells in a 37°C, 5% CO2 incubator for 24 h.

[0147] (c) Detection of cell activity: Dilute the CCK-8 reagent with fresh serum-free culture medium at a volume ratio of 10:1 to prepare a working solution. After 24 hours of intervention with the above-mentioned drug composition, remove the supernatant from each well, add 100 μL of CCK-8 working solution to each well, and continue to incubate in a 37°C cell culture incubator for 1.5 hours before taking out. Set the wavelength parameter of the microplate reader to 450 nm, read the absorbance (OD) value, and use GraphPadPrism v6.0 software to draw a graph and calculate the half-maximal inhibitory concentration (IC) corresponding to 24 hours. 50 ).

[0148] CCK8 assays were used to evaluate the toxic effects of 2-hydroxycinnamic acid and a combination of cinnamic acids on HGC27 cells, which have high levels of EGFRY845 phosphorylation. Figure 15 The results showed that the composition exhibited a growth inhibitory effect on the above-mentioned cell lines, indicating that it has the characteristics of being used to treat tumors in which EGFRY845 phosphorylation is the main driving factor.

[0149] Example 4

[0150] Raw materials: 2-hydroxycinnamic acid and ferulic acid, molar ratio is 1:1.

[0151] 2-Hydroxycinnamic acid and ferulic acid are dissolved in anhydrous ethanol and then mixed to obtain an anti-tumor composition DCF, which is used to treat tumors with high PCNA expression.

[0152] Test Example 4

[0153] CCK-8 pharmacodynamics validation of the antitumor effect of the antitumor composition of Example 4 (on cells with high PCNA levels, AGS)

[0154] Here are the steps:

[0155] (a) 96-well plate plating: After trypsinizing the cells from the cell culture dish and resuspending them, 10 μL of the cell suspension (1:1 dilution) was counted using a cell counting plate and seeded into a 96-well plate at a density of 5000 cells / well. The plate was then placed in a cell culture incubator and allowed to adhere.

[0156] (b) Drug Addition: The next day, after the cells have adhered to the wall, replace each well with 200 μL of fresh serum-free medium. Adjust the final drug concentration to 0, 1.5, and 3 mM. Set up three replicate wells for each concentration group, and include a blank well (without cells) for background subtraction and zeroing. Incubate the cells in a 37°C, 5% CO2 incubator for 24 h.

[0157] (c) Detection of cell activity: Dilute the CCK-8 reagent with fresh serum-free culture medium at a volume ratio of 10:1 to prepare a working solution. After 24 hours of intervention with the above-mentioned drug composition, remove the supernatant from each well, add 100 μL of CCK-8 working solution to each well, and continue to incubate in a 37°C cell culture incubator for 1.5 hours before taking out. Set the wavelength parameter of the microplate reader to 450 nm, read the absorbance (OD) value, and use GraphPadPrism v6.0 software to draw a graph and calculate the half-maximal inhibitory concentration (IC) corresponding to 24 hours. 50 ).

[0158] The CCK8 assay evaluated the toxic effects of a combination of 2-hydroxycinnamic acid and ferulic acid on HGC27 cells with high levels of EGFRY845 phosphorylation. Figure 16 The results showed that the composition exhibited a growth inhibitory effect on the above-mentioned cell lines, indicating that it has the characteristics of being used to treat tumors in which PCNA is the main driving factor.

[0159] Example 5

[0160] Raw materials: 2-hydroxycinnamic acid, cinnamic acid and ferulic acid, with a molar ratio of 1:2:1.

[0161] The preparation method is the same as that of Example 1.

[0162] Test Example 5

[0163] CCK-8 pharmacodynamics verification of the antitumor effect of the antitumor composition of Example 5 (on human gastric cancer HGC27)

[0164] For the test method, see Test Example 3.

[0165] The CCK8 assay evaluated the cytotoxic effects of 2-hydroxycinnamic acid, cinnamic acid, and ferulic acid (molar ratio of 1:2:1) on human gastric cancer HGC27 cells. Figure 17 The results showed that the composition exhibited a good growth inhibitory effect on the above cell lines.

[0166] Example 6

[0167] Raw materials: 2-hydroxycinnamic acid, cinnamic acid and ferulic acid, with a molar ratio of 2:1:1.

[0168] The preparation method is the same as that of Example 1.

[0169] Test Example 6

[0170] CCK-8 pharmacodynamics verification of the antitumor effect of the antitumor composition of Example 6 (on human gastric cancer HGC27)

[0171] For the test method, see Test Example 3.

[0172] The CCK8 assay evaluated the cytotoxic effects of 2-hydroxycinnamic acid, cinnamic acid, and ferulic acid (molar ratio 2:1:1) on human gastric cancer HGC27 cells. Figure 18 The results showed that the composition also exhibited a good growth inhibitory effect on the above cell lines.

[0173] Example 7

[0174] Raw materials: 2-hydroxycinnamic acid, cinnamic acid and methyl ferulate, in a molar ratio of 1:1:1.

[0175] The preparation method is the same as that of Example 1.

[0176] Test Example 7

[0177] CCK-8 pharmacodynamics verification of the antitumor effect of the antitumor composition of Example 7 (on human gastric cancer HGC27)

[0178] For the test method, see Test Example 3.

[0179] The CCK8 assay evaluated the cytotoxic effects of 2-hydroxycinnamic acid, cinnamic acid, and methyl ferulate (molar ratio 1:1:1) on human gastric cancer HGC27 cells. Figure 19 The results showed that the composition also had a good growth inhibitory effect on the above cell lines.

[0180] Example 8

[0181] Raw materials: 2-hydroxycinnamic acid, cinnamic acid and caffeic acid, with a molar ratio of 1:1:1.

[0182] The preparation method is the same as that of Example 1.

[0183] Test Example 8

[0184] CCK-8 pharmacodynamics verification of the antitumor effect of the antitumor composition of Example 8 (on human gastric cancer HGC27)

[0185] For the test method, see Test Example 3.

[0186] The CCK8 assay evaluated the cytotoxic effects of 2-hydroxycinnamic acid, cinnamic acid, and caffeic acid (molar ratio of 1:1:1) on human gastric cancer HGC27 cells. Figure 20 The results showed that the composition also had a good growth inhibitory effect on the above cell lines.

[0187] Test Example 9

[0188] CCK-8 pharmacodynamics validation of the anti-tumor effect of DCF in Example 1

[0189] Here are the steps:

[0190] (a) 96-well plate plating: HGC27 cells were trypsinized and resuspended from the cell culture dish. 10 μL of the cell suspension (1:1 dilution) was counted using a cell counting plate and seeded into 96-well plates at a density of 5000 cells / well. The plates were then placed in a cell culture incubator and allowed to adhere.

[0191] (b) Drug Addition: The next day, after the cells have adhered to the wall, replace each well with 200 μL of fresh serum-free medium. Adjust the final concentration of DCF (from Example 1) to 3 mM and the concentration of cisplatin to 10 μM. Set up three replicate wells for each concentration group, and include a blank well (without cells) for background subtraction and zeroing. The cells were then cultured in a 37°C, 5% CO2 incubator for 24 h.

[0192] (c) Detection of cell activity: Dilute the CCK-8 reagent with fresh serum-free medium at a volume ratio of 10:1 to prepare a working solution. After 24 hours of drug intervention, remove the supernatant from each well, add 100 μL of CCK-8 working solution to each well, and continue to incubate in a 37°C cell culture incubator for 1.5 hours before taking out. Set the wavelength parameter of the microplate reader to 450 nm, read the absorbance (OD) value, and use GraphPad Prism v6.0 software to draw a graph and calculate the half-maximal inhibitory concentration (IC) corresponding to 24 hours. 50 ).

[0193] CCK8 experiment was used to evaluate the cytotoxicity of DCF of Example 1 combined with cisplatin on HGC27. The results are shown in Table 1. Figure 21 The results show that DCF combined with cisplatin shows good synergistic growth inhibition on HGC27 cell line.

[0194] Test Example 10

[0195] CCK-8 pharmacodynamic verification of the anti-tumor effect of DCF of Example 1

[0196] The steps are as follows:

[0197] (a) 96-well plate plating: After trypsin digestion of HGC27 from the cell culture dish and resuspension, 10 μL of cell suspension (1:1 dilution) was counted with a cell counting plate, and was inoculated into a 96-well plate at a density of 5000 cells / well, and then was placed in a cell incubator for adhesion;

[0198] (b) Drug addition: After adhesion culture for one day, 200 μL of fresh serum-free medium was added to each well. The final concentration of DCF of Example 1 was adjusted to 3 (mM), and the concentration of PD-L1 immune checkpoint inhibitor (MDX-1105) was adjusted to 10 (μg / mL), and three parallel wells were set for each concentration group, and a blank well (without cells) was set for background zero reduction, and then was placed in a 37℃, 5% CO2 cell incubator for 24h;

[0199] (c) Detection of cell activity: CCK-8 reagent was diluted with fresh serum-free medium according to a volume ratio of 10:1 to prepare a working solution. After 24h of drug intervention, the supernatant was removed from the wells, and then 100 μL of CCK-8 working solution was added to each well, and was incubated in a 37℃ cell incubator for 1.5h. The wavelength parameter of the microplate reader was set to 450nm, and the absorbance (OD) value was read, and the GraphPad Prismv6.0 software was used to plot and calculate the 24h corresponding half-inhibitory concentration (IC 50 ).

[0200] CCK8 experiment was used to evaluate the cytotoxicity of DCF of Example 1 combined with PD-L1 immune checkpoint inhibitor MDX-1105 on HGC27. The results are shown in Table 1. Figure 22 The results show that DCF combined with cisplatin shows good synergistic growth inhibition on HGC27 cell line.

[0201] Test Example 11

[0202] CCK-8 pharmacodynamic verification of the anti-tumor effect of DCF of Example 1 (on part of EGFR-TKI resistant cell lines H460 and A549)

[0203] Here are the steps:

[0204] (a) 96-well plate plating: After trypsinizing and resuspending the two cell types described above from the cell culture dish, 10 μL of the cell suspension (1:1 dilution) was counted using a cell counting plate and seeded into a 96-well plate at a density of 5000 cells / well. The plate was then placed in a cell culture incubator and allowed to adhere.

[0205] (b) Drug Addition: The next day, after the cells have adhered to the wall, replace each well with 200 μL of fresh serum-free medium. Adjust the final concentration of DCF from Example 1 to 0 and 3 mM. Set up three replicate wells for each concentration group, and include a blank well (without cells) for background subtraction and zeroing. Then, incubate the cells in a 37°C, 5% CO2 cell culture incubator for 24 h.

[0206] (c) Detection of cell activity: Dilute the CCK-8 reagent with fresh serum-free medium at a volume ratio of 10:1 to prepare a working solution. After 24 hours of drug intervention, remove the supernatant from each well, add 100 μL of CCK-8 working solution to each well, and continue to incubate in a 37°C cell culture incubator for 1.5 hours before taking out. Set the wavelength parameter of the microplate reader to 450 nm, read the absorbance (OD) value, and use GraphPad Prism v6.0 software to draw a graph and calculate the half-maximal inhibitory concentration (IC) corresponding to 24 hours. 50 ).

[0207] The CCK8 assay evaluated the cytotoxic effects of DCF (cocktail and cocktail liposomes, i.e., DCF and DCF in a liposome nanostructured context) of Example 1 on EGFR-TKI-resistant cell lines (H460 and A549). Figure 23 The results showed that the DCF (cocktail and cocktail liposomes) of Example 1 exhibited growth inhibitory effects on both of the above-mentioned cell lines, indicating that it has potential to overcome drug resistance.

[0208] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. An antitumor composition, characterized in that: comprising a first component and a second component; The first component includes 2-hydroxycinnamic acid; The second component includes ferulic acid and / or cinnamic acid; The molar ratio of the first component to the second component is (1-2): (1-3).

2. The antitumor composition according to claim 2, characterized in that The anti-tumor composition comprises the following components: ferulic acid, 2-hydroxycinnamic acid and cinnamic acid.

3. The antitumor composition according to claim 2, characterized in that The molar ratio of the ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is (1-2): (1-2):

1.

4. The antitumor composition according to claim 3, characterized in that The molar ratio of the ferulic acid, 2-hydroxycinnamic acid and cinnamic acid is 1:1:

1.

5. The antitumor composition according to any one of claims 1 to 4, characterized in that The ferulic acid is replaced by methyl ferulate or caffeic acid.

6. An anti-tumor drug delivery system, characterized in that: The active ingredient comprises the anti-tumor composition according to any one of claims 1 to 5.

7. The anti-tumor drug delivery system according to claim 6, characterized in that The anti-tumor drug delivery system includes a liposome delivery system; The liposome delivery system comprises the following raw materials: lecithin, cholesterol, the antitumor composition according to any one of claims 1 to 5, and a solvent; The mass ratio of the lecithin, cholesterol and anti-tumor composition is 5:1:

1.

8. Use of the antitumor composition according to any one of claims 1 to 5 or the antitumor drug delivery system according to claim 6 or 7 in the preparation of antitumor drugs.

9. The use according to claim 8, characterized in that The tumor includes a tumor with phosphorylation of EGFRY845 and / or high expression of PCNA.

10. The use according to claim 8 or 9, characterized in that: The tumor includes at least one of gastric cancer, lung cancer and breast cancer.

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