Anti-tumor and / or Anti-ferroptosis small molecule composition and use thereof

Abstract

A small molecule composition, comprising licochalcone A, licochalcone C, licochalcone D, licochalcone E, echinatin, isoliquiritigenin, formononetin, liquiritigenin, licoflavone C, glabrol, licoisoflavone A and licoisoflavone B, and the use thereof in preparing anticancer or antitumor drugs, anti-ferroptosis drugs, or drugs for sensitization of anticancer or antitumor drugs. The small molecule composition can also be used in combination with other antitumor drugs.

Description

Small molecule compositions with antitumor and / or antiferroptosis and their applications Technical Field

[0001] This invention belongs to the field of pharmaceutical technology. Specifically, this invention provides a small molecule composition that can be used to prevent or treat diseases such as tumors, especially in combination with other anti-tumor drugs to prevent or treat diseases such as tumors, and can also be used to inhibit ferroptosis of cells. Background Technology

[0002] Cancer threatens human life and health. Although some progress has been made after more than half a century of efforts, the etiology and pathology of tumors are extremely complex. Factors such as the heterogeneity, clonal nature, and adaptive variability of tumor cells make it difficult for various anticancer drugs to maintain their effectiveness for a long time. Ultimately, they all fail due to tumor drug resistance.

[0003] Ferroptosis is a programmed cell death process caused by the accumulation of iron-dependent lipid peroxidation products. When the activity of glutathione peroxidase 4 (GPX4), a key regulator of ferroptosis, is inhibited, the cell's antioxidant capacity decreases, leading to a large accumulation of reactive oxygen species (ROS), which in turn triggers oxidative damage and causes ferroptosis. Ferroptosis is widely present in the process of organ and tissue decay.

[0004] Licorice is a common Chinese herbal medicine. In traditional Chinese medicine prescriptions, licorice is usually used in decoctions. From a pharmacological perspective, most of its components are water-soluble molecules. Licorice contains numerous components, and the interactions between these components are quite complex.

[0005] The inventors have long been committed to the modernization research of traditional Chinese medicine. In the early stages, they developed Z018A from licorice extract, which is disclosed in Chinese Patent ZL201610169121.8 (the full text of which is included in this article for reference). Z018A has three characteristic peaks on the HPLC detection chromatogram (expected to contain only three components), and can be used to prevent or treat fatty liver damage and obesity, reduce blood lipid levels, and reduce inflammatory factor levels.

[0006] Based on this, the inventors further developed a crystalline product of Z018A, disclosed in Chinese Patent ZL201910410704.9 (the full text of which is incorporated herein by reference). It also exhibits three characteristic peaks on its HPLC chromatogram (expected to contain only three components), and is a SIRT1 agonist that can significantly increase SIRT1 expression levels in vivo, potentially useful for anti-aging.

[0007] Research on Z018A subsequently ceased. However, the inventors did not give up. In a less-than-successful experiment while studying the extraction of Z018A, they unexpectedly discovered a licorice extract component, Z018C, and modernized and improved it, developing a small molecule drug composition with a clear composition and stable quality. This composition can inhibit tumors (including drug-resistant tumors) alone or in combination with existing antitumor drugs. In particular, it exhibits a very significant synergistic effect when used in combination with commonly used anticancer drugs, and it also has a protective effect against ferroptosis. Summary of the Invention

[0008] The technical problem to be solved by this invention is to provide novel small molecule compositions that can be used for the prevention or treatment of diseases such as tumors, especially in combination with other anti-tumor drugs for the prevention or treatment of diseases such as tumors, and can also be used to inhibit ferroptosis of cells. In addition, this invention also provides pharmaceutical formulations comprising this small molecule composition and its applications.

[0009] Specifically, in a first aspect, the present invention provides a small molecule composition comprising glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, monigrin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A, and glycyrrhizin isoflavone B. The composition of the first aspect of the present invention has clearly defined components. In specific embodiments of the present invention, it is preferred that the composition of the first aspect of the present invention comprises glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, monigrin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A, and glycyrrhizin isoflavone B.

[0010] The small molecule composition of the first aspect of the present invention can be used for anti-cancer or anti-tumor purposes; that is, the small molecule composition provided by the present invention can be an anti-cancer or anti-tumor composition.

[0011] The small molecule composition of the first aspect of this invention can also be used to combat ferroptosis; that is, the small molecule composition provided by this invention can also be an antiferroptosis composition. Antiferroptosis compositions can be used to combat symptoms such as organ damage.

[0012] The small molecule composition of the first aspect of this invention can also be used for sensitizing anticancer or tumor drugs, that is, increasing the sensitivity of cancer or tumor to anticancer or tumor drugs. Specifically, the small molecule composition provided by this invention can also be a sensitizing composition for anticancer or tumor drugs. Numerous anticancer or tumor drugs are currently available. Preferably, in this invention, the anticancer or tumor drug is an EGFR-targeting anticancer or tumor drug (e.g., osimertinib) and / or a cytotoxic anticancer or tumor drug (preferably 5-fluorouracil or platinum compounds, such as oxaliplatin or cisplatin). In a specific embodiment of this invention, the anticancer or tumor drug targeted for sensitization is osimertinib and / or 5-fluorouracil.

[0013] In this invention, glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, gentiopicrin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A and / or glycyrrhizin isoflavone B can be extracted from plants or synthesized artificially. Preferably, the plant is a plant of the genus Glycyrrhiza, such as licorice (Glycyrrhiza uralensis Fisch.), squamulosa (Glycyrrhiza squamulosa Franch.), inflata (Glycyrrhiza inflata Batalin), pallidiflora (Glycyrrhiza pallidiflora Maxim.), glabra (Glycyrrhiza glabra L.), or aspera (Glycyrrhiza aspera Pall.), etc.

[0014] Preferably, in the composition of the first aspect of the present invention, the weight ratio of glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, gentianin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A and glycyrrhizin B is 6.2~6.8:0.62~0.68:0.4~0.44:0.83~0.92:0.1~0.13:0.12~0.18:0.4~0.5:0.18~0.25:0.1~0.13:0.15~0.2:0.16~0.21:0.1~0.15.

[0015] More preferably, in the composition of the first aspect of the present invention, the weight ratio of glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, gentianin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A and glycyrrhizin B is 6.4~6.6: 0.64~0.66: 0.41~0.43: 0.87~0.89: 0.11~0.13: 0.14~0.16: 0.44~0.46: 0.2~0.22: 0.11~0.13: 0.17~0.19: 0.18~0.2: 0.12~0.14.

[0016] In a specific embodiment of the present invention, the weight ratio of glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, gentianin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A and glycyrrhizin B is 6.5:0.65:0.42:0.88:0.12:0.15:0.45:0.21:0.12:0.18:0.19:0.13.

[0017] In a second aspect, the present invention provides an anticancer or tumor composition comprising the composition of the first aspect of the present invention and an anticancer or tumor drug.

[0018] There are numerous types of cancer or tumors. Preferably, in this invention, the cancer or tumor can be a lung tumor, liver tumor, stomach tumor, colon cancer, and / or pancreatic cancer.

[0019] There are numerous existing anticancer or tumor drugs. Preferably, in this invention, the anticancer or tumor drug is an EGFR-targeting anticancer or tumor drug (e.g., osimertinib) and / or a cytotoxic anticancer or tumor drug (preferably 5-fluorouracil or platinum compounds, such as oxaliplatin or cisplatin).

[0020] Preferably, in this invention, the cancer or tumor is a drug-resistant cancer or tumor, for example, it may be a cancer or tumor resistant to EGFR-targeted anticancer or tumor drugs (such as osimertinib).

[0021] In a third aspect, the present invention provides a formulation comprising the composition of the first or second aspect of the present invention and pharmaceutically acceptable excipients. The formulation may be a pharmaceutical preparation.

[0022] In this document, the term "pharmaceuticalally acceptable excipient" includes pharmaceutically acceptable carriers, excipients, diluents, etc., which are compatible with the active pharmaceutical ingredient. The use of pharmaceutically acceptable excipients in the preparation of pharmaceutical formulations is well known to those skilled in the art. The pharmaceutical formulations of this invention comprise the compositional components of the first or second aspect of this invention as the active ingredient, combined with pharmaceutically acceptable excipients (such as carriers, excipients, diluents, etc., well known to those skilled in the art), to formulate various formulations, preferably solid and liquid formulations, such as tablets, pills, capsules (including sustained-release or delayed-release forms), powders, suspensions, granules, syrups, emulsions, and suspensions, as well as various sustained-release formulations, preferably for oral administration. In one specific embodiment of this invention, the composition of the first or second aspect of this invention is diluted with a solution to form a liquid formulation. The effective amount for prevention or treatment based on the composition of the first or second aspect of this invention can be estimated from the dosage in experimental animals.

[0023] In a fourth aspect, the present invention provides the use of the composition of the first aspect of the present invention in the preparation of a medicament for treating cancer or tumors. Accordingly, in a fifth aspect, the present invention provides a method for treating cancer or tumors, comprising administering the composition of the first aspect of the present invention.

[0024] The application of the fourth aspect or the method of the fifth aspect of the present invention can be the sole application or method of the composition of the first aspect of the present invention, or it can be the application or method in combination with other anticancer or tumor drugs. Preferably, the application of the fourth aspect or the method of the fifth aspect of the present invention is the application or method of the composition of the first aspect of the present invention in combination with anticancer or tumor drugs.

[0025] In a sixth aspect, the present invention provides the use of the composition of the first aspect of the present invention in the preparation of a medicament for inhibiting ferroptosis. Accordingly, in a seventh aspect, the present invention provides a method for inhibiting ferroptosis, comprising administering the composition of the first and second aspects of the present invention. In specific embodiments of the present invention, the application of the sixth aspect or the method of the seventh aspect is either the application or method of the composition of the first aspect of the present invention alone.

[0026] In an eighth aspect, the present invention provides the use of the composition of the first aspect of the invention in the preparation of a medicament for sensitizing anticancer or tumor drugs. Accordingly, in a ninth aspect, the present invention provides a method for sensitizing anticancer or tumor drugs, comprising administering the composition of the first aspect of the invention. Anticancer or tumor drugs often have significant side effects; therefore, sensitization can reduce the dosage of anticancer or tumor drugs used and alleviate side effects. The composition of the first aspect of the present invention can individually increase the sensitivity of cancer or tumors to anticancer or tumor drugs. In a specific embodiment of the present invention, the anticancer or tumor drug is osimertinib and / or 5-fluorouracil.

[0027] In this invention, the application subject can be a human or an experimental animal, preferably a human.

[0028] The beneficial effects of this invention are that it provides a novel small molecule composition that can be used as an anticancer or tumor-fighting composition, as an antiferroptosis composition, or as an anticancer or tumor drug sensitizer composition. In particular, the small molecule composition of this invention exhibits a very significant synergistic anticancer effect when used in combination with commonly used anticancer drugs, especially increasing the sensitivity of drug-resistant tumors to already resistant drugs and enhancing the antitumor effect.

[0029] For ease of understanding, the present invention will be described in detail below with reference to specific embodiments and accompanying drawings. It should be particularly noted that these descriptions are merely exemplary and do not constitute a limitation on the scope of the invention. Many variations and modifications of the invention will be apparent to those skilled in the art based on the discussion in this specification. Attached Figure Description

[0030] Figure 1 shows the characteristic spectrum of the Z018C extract.

[0031] Figure 2 shows the chemical structure of the main components in the Z018C extract.

[0032] Figure 3 shows the tumor-inhibiting effects of different dose groups of Z018C and the positive control group after 31 days of treatment.

[0033] Figure 4 shows the effect of Z018C dosage groups and the positive control group on animal body weight after 31 days of treatment.

[0034] Figure 5 shows the antitumor effect of Z018C combined with osimertinib in HCC827 cells.

[0035] Figure 6 shows the antitumor effect of Z018C combined with osimertinib in Osimertinib-R-HCC827 cells.

[0036] Figure 7 shows the antitumor effect of Z018C combined with osimertinib in A549 cells.

[0037] Figure 8 shows the antitumor effect of the combination of Z018C and 5-Fu in HCT116 cells.

[0038] Figure 9 shows the protective effect of Z018C against ferroptosis. Detailed Implementation

[0039] The following examples further illustrate the content of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art and commonly used commercially available instruments and reagents. For reference, please refer to the manufacturer's instructions for the corresponding instruments and reagents.

[0040] Example 1: Preparation of Z018C Extract

[0041] The roots and rhizomes of plants in the genus *Glycyrrhiza* were pulverized, extracted by reflux with ethanol, and the extracts were combined. The ethanol was recovered and concentrated to a suitable volume, then added to a pre-treated macroporous adsorption resin column. The column was eluted sequentially with ethanol of different concentrations, and the eluents were collected. The ethanol was recovered and concentrated to a suitable volume, then added to a pre-treated polyamide resin column. The eluents were eluted sequentially with ethanol of different concentrations, and the eluents were collected. The ethanol was recovered, and the column was concentrated into a thick paste. The paste was then dried under reduced pressure to obtain the final product. Among these, component Z018C (the extract) exhibited properties of interest in the preliminary experiments, and therefore warranted further investigation.

[0042] Example 2 Identification of Z018C extract

[0043] The HPLC characteristic chromatogram of the Z018C extract is shown in Figure 1. The main components are shown in Figure 2 after identification and structural analysis.

[0044] Example 3 Preparation of Z018C composition

[0045] To ensure product stability, the following ingredients were prepared according to the proportions of the main components in the Z018C extract: 6.5g of glycyrrhizin A, 0.65g of glycyrrhizin C, 0.42g of glycyrrhizin D, 0.88g of glycyrrhizin E, 0.12g of prickly licorice chalcone, 0.15g of isoglycyrrhizin, 0.45g of gentianin, 0.21g of glycyrrhizin, 0.12g of glycyrrhizin C, 0.18g of glycyrrhizinol, 0.19g of glycyrrhizin A, and 0.13g of glycyrrhizin B. After thorough mixing, this constituted the Z018C composition, which was used for further testing.

[0046] Example 4: Z018C can inhibit the viability of various human tumor cell lines in a time- and dose-dependent manner.

[0047]

Experimental Materials

[0048] 1. Cell lines and culture methods are shown in Table 1.

[0049] Table 1. Cell lines and culture methods

[0050]

[0051] 2. Culture media and reagents, see Table 2.

[0052] Table 2. Culture media and reagents

[0053]

[0054] 3. Experimental apparatus, see Table 3.

[0055] Table 3. Experimental Apparatus

[0056]

[0057] 4. Drugs to be tested and those that test positive

[0058] Test drug: Z018C composition.

[0059] Positive drug: Staurosporine.

[0060] 5. Reagents and instruments used in cell viability assays

[0061] Greiner CELLSTAR 96 - Perforated plate, flat black plate (with cover and transparent bottom), # 655090.

[0062] Promega CellTiter-Glo luminescent cell viability assay kit (Promega-G7573).

[0063] 2104 EnVision board reader.

[0064]

Experimental Methods

[0065] 1. Cell Culture

[0066] The tumor cell lines were cultured in an incubator at 37ºC and 5% CO2 according to the culture conditions shown in Table 1. Cells were passaged periodically, and cells in the logarithmic growth phase were used for plating.

[0067] 2. Cell plating

[0068] (1) Use trypan blue to stain cells and count live cells.

[0069] (2) Adjust the cell concentration to a suitable level, see Table 4.

[0070] Table 4. Cell plating density

[0071]

[0072] (3) Add 90 μL of cell suspension to each well of the culture plate. Control: solvent control, only cells, culture medium, and solvent (DMSO) are added, no drug is added; Blank: blank control, only cell culture medium is added, no cells; Drug experimental group: serially diluted (final concentration from 50 to 0.390625 μg / mL) of the test drug (dissolved in DMSO) is added; Positive drug group: positive drug is added to 1 μM. Incubate the culture plates at 37°C, 5% CO2, and 100% relative humidity for 48 hours or 72 hours, respectively.

[0073] 3. CellTiter-Glo luminescence assay for cell viability detection

[0074] Follow the instructions for the Promega CellTiter-Glo luminescence assay kit (Promega-G7573).

[0075]

Experimental Results

[0076] The anti-cell proliferation parameters obtained from the cell proliferation inhibition experiment are shown in Tables 5 and 6, including minimum inhibition rate, maximum inhibition rate, and IC50. 50 .

[0077] Table 5. Anti-cell proliferation parameters after 48 h of drug treatment

[0078]

[0079] Table 6. Anti-cell proliferation parameters after 72 h of drug treatment

[0080]

[0081]

Experimental Conclusion

[0082] This study investigated the inhibitory effect of Z018C on the proliferation of 16 human tumor cell lines using in vitro anti-proliferation assays. The results showed that Z018C could inhibit the viability of various human tumor cell lines in a time- and dose-dependent manner, significantly inhibiting the cell viability of human lung tumor cell lines A549, NCI-H1299, HCC827, human liver tumor cell lines Huh7, HepG2, SK-HEP-1, human gastric tumor cell lines AGS, MKN45, SUN-1, human colon cancer cell lines WiDr, C2BBe1, HCT116, and human pancreatic cancer cell lines Panc-1, MiaPaCa-2, and BxPC-3.

[0083] Example 5: Mouse xenograft model of lung cancer HCC827 cells

[0084]

Experimental Methods

[0085] 1. Cell Culture: Human lung cancer HCC827 cells (ATCC-CRL-2868) were cultured in vitro in a monolayer under the following conditions: RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution, incubated at 37 ºC in a 5% CO2 incubator. Cells were passaged twice a week using trypsin-EDTA digestion. When cell saturation reached 80%-90% and the desired number was achieved, cells were harvested, counted, and seeded.

[0086] 2. Animals: Female BALB / c nude mice, 6-8 weeks old, weighing 18-20 grams (animal weight subject to actual conditions). A total of 45 mice are required (30 mice + 50% surplus). Supplied by a qualified supplier.

[0087] 3. Tumor inoculation: 10x10 6 One HCC827 cell (0.2 mL, PBS:Matrigel = 1:1) was subcutaneously inoculated into the right back near the upper limb of each mouse, and the average tumor volume reached approximately 150 mm. 3 The drug efficacy experiment will begin with group dosing. The experimental groups and dosing regimens are shown in Table 7 below.

[0088] Table 7 Animal experimental grouping and dosing regimen

[0089]

[0090] Note: 1. N: Number of mice per group 2. Dosage volume: 10 µL / g based on mouse body weight.

[0091] 4. Experimental indicators: Tumor diameter was measured twice a week using vernier calipers. The formula for calculating tumor volume is: V = 0.5 x axb 2 , where a and b represent the long and short diameters of the tumor, respectively. The antitumor efficacy of the test substance was evaluated using the tumor growth inhibition rate (TGI) (%), the relative tumor proliferation rate (T / C) (%), or the tumor regression rate (TR) (%). The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the beginning of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the beginning of treatment in the solvent control group)] × 100 (%).

[0092] Calculation of T / C (%): T / C (%) = T RTV / C RTV × 100( %) (T RTV Treatment group RTV; C RTV (Soluble control group RTV). Relative tumor volume (RTV) is calculated based on tumor volume measurements using the formula: RTV = V t / T0, where T0 is the average tumor volume measured at the time of grouped drug administration (i.e., D0), Vt is the average tumor volume at a certain measurement, and T RTV With C RTV Take data from the same day.

[0093] 5. Data Analysis: The t-test is used for comparisons between two groups. One-way ANOVA is used for comparisons between three or more groups. If the F-values ​​are statistically significant, multiple comparisons should be performed after ANOVA analysis. All data analyses were performed using GraphPad Prism 9. A p-value < 0.05 was considered statistically significant.

[0094]

Experimental Results

[0095] Experimental results showed that Z018C inhibited tumor growth in mice in a time- and dose-dependent manner (see Figure 3). Although the effect was worse than that of the positive control drug osimertinib, Z018C was better tolerated in animals, and no significant weight loss was observed (see Figure 4).

[0096] Example 6: Anticancer Experiment with Z018C Combination

[0097]

Experimental Materials

[0098] 1. Cell lines and culture methods are shown in Table 8.

[0099] Table 8. Cell lines and culture methods

[0100]

[0101] 2. Culture media and reagents are shown in Table 9.

[0102] Table 9. Culture media and reagents

[0103]

[0104] 3. Test and positive drugs

[0105] Drug to be tested 1: Z018C

[0106] Drug to be tested 2: Osimertinib

[0107] Test drug 3: 5-Fu (5-fluorouracil)

[0108] 4. Instruments

[0109] Greiner CELLSTAR 96-perforated board, flat black board (with cover and transparent bottom), # 655090.

[0110] 5. Reagents and instruments used in cell viability experiments

[0111] Promega CellTiter-Glo chemiluminescence immunoassay kit (Promega-G7573).

[0112] 2104 EnVision board reader, PerkinElmer.

[0113] Tecan D300e micro-volume pipette, Hewlett-Packard.

[0114]

Experimental Methods and Procedures

[0115] 1. Cell Culture

[0116] The tumor cell lines were cultured in incubators at 37ºC and 5% CO2. Cells were passaged periodically, and those in the logarithmic growth phase were used for plating.

[0117] 2. Cell plating

[0118] (1) Use trypan blue to stain cells and count live cells.

[0119] (2) Adjust the cell concentration to a suitable level.

[0120]

[0121] (3) Add 100 μL of cell suspension to each well of the culture plate and add cell-free culture medium to the blank control well.

[0122] (4) Incubate the culture plate overnight in an incubator at 37°C, 5% CO2 and 100% relative humidity.

[0123] 3. Add each test drug and its mixture to the wells of the plate at serially diluted concentrations. Control: solvent control, only cells, culture medium, and solvent are added, no drug is added; Blank: blank control, only cell culture medium is added, no cells are added; For HCC827, the drug experimental group: serially diluted test drug 1 (final concentrations of 25 to 0.1953 μg / mL and 0, and test drug 2 (final concentrations of 0.1000 to 0.00005 μM and 0, respectively) is added; For Osimer For tinib-R-HCC827 and A549, serially diluted test drugs (final concentrations of 25 to 0.1953 μg / mL and 0 for test drug 1, and final concentrations of 10.0000 to 0.00457 μM and 0 for test drug 2) were added. For HCT116, in the drug experimental group, serially diluted test drugs (final concentrations of 25 to 0.1953 μg / mL and 0 for test drug 1, and final concentrations of 100.0000 to 0.7813 μM and 0 for test drug 3) were added. The 96-well cell plates were then incubated for 72 h. Cell viability was assessed using CellTiter-Glo chemiluminescence assay.

[0124] 4. Data Analysis

[0125] The inhibition rate (IR) of the detected compound is calculated using the following formula: IR (%) = [1 – (RLU compound – RLU blank control) / (RLU solvent control – RLU blank control)]*100%. The inhibition rates of different compound concentrations are calculated in Excel, and then inhibition curves are plotted and relevant parameters, including minimum inhibition rate, maximum inhibition rate, and IC50, are calculated using GraphPad Prism software. 50 .

[0126]

Experimental Results

[0127] 1. Z018C in combination with osimertinib:

[0128] (1) In EGFR mutation-positive HCC827 cells, low dose (3.125 μg / mL) Z018C significantly improved the sensitivity of osimertinib, and the inhibitory effect on tumor growth was about 160% higher than that of osimertinib alone, suggesting that Z018C and osimertinib are an effective combination for the treatment of EGFR mutation-positive non-small cell lung cancer. The results are shown in Figure 5.

[0129] (2) In the Osimertinib-R-HCC827 (Osimertinib-resistant HCC827) cell model, Z018C can reduce osimertinib resistance in a dose-dependent manner, increase the sensitivity of resistant cells to osimertinib, and increase the inhibitory effect on tumor growth by about 300% compared with osimertinib alone. This suggests that the combination of Z018C and osimertinib shows a significant sensitizing effect in the treatment of osimertinib-resistant non-small cell lung cancer cell lines. The results are shown in Figure 6.

[0130] (3) In the A549 cell model, Z018C dose-dependently increased the sensitivity of osimertinib, and the inhibitory effect on tumor growth was up to 300% higher than that of osimertinib alone. This suggests that Z018C has good anti-tumor activity against non-small cell lung cancer with non-driver gene mutations that are resistant to osimertinib, and significantly enhances the anti-tumor activity of osimertinib. The results are shown in Figure 7.

[0131] In summary, Z018C is effective against both non-driver gene mutation and driver gene mutation non-small cell lung cancer. When used in combination with osimertinib, it can enhance efficacy, reduce toxicity, delay drug resistance, and improve quality of life.

[0132] 2. Z018C in combination with 5-fluorouracil (5-Fu):

[0133] In the HCT116 cell model, Z018C significantly enhanced the tumor-suppressing effect of 5-fluorouracil compared to 5-fluorouracil alone, suggesting that Z018C and 5-fluorouracil are an effective combination for treating colorectal cancer (see Figure 8).

[0134] Example 7: Ferraphobia Sensitivity Analysis Based on Cell Viability

[0135]

Experimental Materials

[0136] 1. Cell line: HT1080.

[0137] 2. Drug to be tested: Z018C.

[0138]

Experimental Methods and Procedures

[0139] 1. Cell Culture

[0140] The HT1080 cell line was cultured in an incubator at 37ºC with 5% CO2. Cells were passaged periodically, and those in the logarithmic growth phase were used for plating.

[0141] 2. Cell plating

[0142] Add 100 μL of cell suspension (10) to each well of the culture plate. 5(1 cell / well), and add cell-free culture medium to the blank control wells. Incubate the culture plate in an incubator at 37°C, 5% CO2, and 100% relative humidity for 24 hours.

[0143] 3. The test drug was added to five wells at concentration gradients of 0, 1, 3, 9, and 27 μg / ml. The blank control group received only DMSO solvent, without the drug. The tolerance test group received serially diluted test drug. The experimental group received serially diluted test drug and a final concentration of 0.05 μM of the GPX4 (glutathione peroxidase 4) inhibitor RSL3. The culture plates were then incubated for 12 hours. Cell viability was assessed using a CCK-8 assay.

[0144]

Experimental Results

[0145] As shown in Figure 9, for HT1080 cells, the concentration tolerance range of Z018C is ≤9 μg / ml. Within the concentration tolerance range, Z018C can reduce the sensitivity of cells to ferroptosis and has a protective effect against ferroptosis. Moreover, the protective effect gradually increases with the increase of Z018C concentration.

Claims

1. A small molecule composition comprising glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, gentiopicrin, glycyrrhizin, glycyrrhizin C, glycyrrhizin, glycyrrhizin isoflavone A and glycyrrhizin B.

2. The composition of claim 1, wherein, The weight ratio of glycyrrhizin A, glycyrrhizin C, glycyrrhizin D, glycyrrhizin E, prickly glycyrrhizin, isoglycyrrhizin, gentianin, glycyrrhizin, glycyrrhizin C, glycyrrhizinol, glycyrrhizin isoflavone A, and glycyrrhizin isoflavone B is 6.2~6.8:0.62~0.68:0.4~0.44:0.83~0.92:0.1~0.13:0.12~0.18:0.4~0.5:0.18~0.25:0.1~0.13:0.15~0.2:0.16~0.21:0.1~0 0.15, preferably 6.4~6.6:0.64~0.66:0.41~0.43:0.87~0.89:0.11~0.13:0.14~0.16:0.44~0.46:0.2~0.22:0.11~0.13:0.17~0.19:0.18~0.2:0.12~0.14, most preferably 6.5:0.65:0.42:0.88:0.12:0.15:0.45:0.21:0.12:0.18:0.19:0.

13.

3. The composition of claim 1, used for anticancer or tumor treatment, for anti-ferroptosis, or for sensitizing anticancer or tumor drugs.

4. An anticancer or tumor composition comprising the composition of claim 1 or 2 and an anticancer or tumor drug.

5. The composition of claim 4, wherein, The anticancer or tumor drugs are EGFR-targeted anticancer or tumor drugs (e.g., osimertinib) and / or cytotoxic anticancer or tumor drugs (preferably 5-fluorouracil or platinum compounds such as oxaliplatin or cisplatin).

6. The composition of claim 4, wherein, The cancer or tumor is a drug-resistant cancer or tumor, preferably a cancer or tumor resistant to EGFR-targeted anticancer or tumor drugs (e.g., osimertinib).

7. A formulation comprising the composition according to any one of claims 1 to 6 and pharmaceutically acceptable excipients.

8. Use of the composition according to any one of claims 1 to 6 in the preparation of a medicament for anticancer or tumor treatment, for anti-ferroptosis treatment, or for sensitizing anticancer or tumor drugs.

9. The use of claim 8, wherein, The cancer or tumor is a lung tumor, liver tumor, stomach tumor, colon cancer and / or pancreatic cancer, or the cancer or tumor is a drug-resistant cancer or tumor, preferably a cancer or tumor resistant to EGFR-targeted anticancer or tumor drugs (e.g., osimertinib).

10. The application according to claim 8, wherein the composition according to any one of claims 1 to 6 is used in combination with an anticancer or tumor drug.

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