Novel her2 inhibitors and methods of making and using the same

Abstract

The application provides a novel HER2 inhibitor and a preparation method and application thereof. The novel HER2 inhibitor is a small molecule compound, and a structural formula of the small molecule compound is shown as formula 1. The application selects NCI-N87 cells as a HER2 positive gastric cancer cell model to verify the targeting effect of the compound shown as formula 1 on a HER2 signal pathway and the antitumor activity. The experimental results show that the compound shown as formula 1 has the targeting effect on HER2, can inhibit HER2 positive tumors, and is expected to become a new generation of antitumor drugs.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to novel HER2 inhibitors, their preparation methods, and applications. Background Technology

[0002] Human epidermal growth factor receptor 2 (HER2) is associated with the development and progression of various cancers, including gastric cancer and breast cancer, and is one of the important targets for targeted cancer therapy.

[0003] The mechanisms of action of existing anti-HER-2 drugs include: targeting the HER-2 receptor: HER-2 (human epidermal growth factor receptor 2) is a transmembrane protein that promotes cell proliferation, and overexpression can lead to malignant tumor progression; blocking signaling pathways: by binding to the HER-2 receptor or inhibiting its activity, they interfere with downstream pro-cancer signaling (such as the PI3K / AKT and MAPK pathways); activating immune killing: some drugs enhance the immune system's clearance of tumor cells through antibody-dependent cytotoxicity (ADCC).

[0004] The main types of drugs and representative drugs include monoclonal antibodies (such as trastuzumab and pertuzumab), small molecule tyrosine kinase inhibitors (such as small molecule tyrosine kinase inhibitors), antibody-drug conjugates, and bispecific antibodies (such as maggotuximab, which targets both HER-2 and CD3, activating T cells to kill tumors). While monoclonal antibodies have the advantages of high specificity and potency, precisely targeting the HER2 receptor, their large molecular weight leads to low oral bioavailability, requiring injection administration. Furthermore, monoclonal antibodies are expensive to produce and can easily cause immune and allergic reactions during treatment. Currently approved HER2 small molecule inhibitors still face challenges in clinical application, such as drug resistance, insufficient targeting specificity, and side effects. Therefore, developing new HER2 small molecule inhibitors has become a current research focus. Summary of the Invention

[0005] One of the objectives of this invention is to provide a compound.

[0006] The compound provided by this invention has the structural formula shown in Formula 1:

[0007]

[0008] The compound shown in Formula 1 above, according to Figure 1 The flowchart shown is prepared by a method including the following steps:

[0009] 1) Compound 1 and compound 2 react under alkaline conditions in an organic solvent to give compound 3;

[0010]

[0011] 2) Compound 3 undergoes a reduction reaction in the presence of a reducing agent to give compound 4;

[0012]

[0013] 3) Compound 4 reacts with bromoacetonitrile (BrCH2CN) to give compound 6;

[0014]

[0015] 4) Compound 6 and compound 7 react in the presence of a palladium catalyst and a cesium salt to give the target compound.

[0016] .

[0017] In step 1) of the above method, the alkali is potassium carbonate;

[0018] The molar ratio of compound 1 to compound 2 can be 1:1 to 1:2;

[0019] The reaction temperature can be 60-100°C and the time can be 12-20 h, specifically 16 h at 80°C.

[0020] In step 2) of the above method, the reducing agent may specifically be sodium borohydride.

[0021] The molar ratio of compound 3 to sodium borohydride can be 1:2-3, specifically 1:2;

[0022] The reduction reaction can be carried out at room temperature for 2-5 hours, specifically 3 hours.

[0023] In step 3) of the above method, the ratio of compound 4 to bromoacetonitrile is 1:4-6, specifically 1:5;

[0024] The reaction is carried out under alkaline conditions, and the alkaline substance may be NaH.

[0025] The reaction can be carried out at room temperature for 10-16 hours, specifically 12 hours.

[0026] In step 4) of the above method, the palladium catalyst can specifically be Pd(dppf)Cl2;

[0027] The cesium salt may specifically be cesium carbonate;

[0028] The molar ratios of compound 6 to compound 7, palladium catalyst, and cesium salt can be 1:2-3:0.1-0.2:2-3, respectively.

[0029] The reaction is carried out in an inert atmosphere;

[0030] The reaction temperature can be 80-120 °C, specifically 100 °C, and the time can be 8-16 h, specifically 12 h.

[0031] Another object of the present invention is to provide the use of the compound shown in Formula 1 in the preparation of HER2-positive tumor diagnostic reagents.

[0032] The specific application may be the use of the compound shown in Formula 1 in the preparation of drugs for the prevention and / or treatment of HER2-positive tumors.

[0033] The tumor is cancer, specifically breast cancer, lung cancer, stomach cancer, or colorectal cancer.

[0034] In the aforementioned application, the drug has at least one of the following effects:

[0035] 1) Inhibits the proliferation of tumor cells;

[0036] 2) Inhibits tumor cell survival and clone formation;

[0037] 3) Blocking of the tumor cell cycle.

[0038] The present invention also provides a medicament for the prevention and / or treatment of HER2-positive tumors, the medicament comprising a compound shown in Formula 1.

[0039] The tumor is cancer, specifically breast cancer, lung cancer, stomach cancer, or colorectal cancer.

[0040] This invention selected NCI-N87 cells as a HER2-positive gastric cancer cell model to verify the targeting effect and anti-tumor activity of the compound shown in Formula 1 on the HER2 signaling pathway. Experimental results show that the compound shown in Formula 1 has a targeting effect on HER2 and can inhibit HER2-positive tumors, potentially becoming a next-generation anti-tumor drug. Attached Figure Description

[0041] Figure 1 This is a flowchart illustrating the preparation process of the compound represented by Formula 1 in this invention.

[0042] Figure 2 The image shows the hydrogen NMR spectrum of the compound of Formula 1 prepared in Example 1 of this invention.

[0043] Figure 3 The mass spectrum of the compound of Formula 1 prepared in Example 1 of this invention is shown.

[0044] Figure 4 The HPLC purity results are for the compound of Formula 1 prepared in Example 1 of this invention.

[0045] Figure 5 In Example 2 of this invention, the effect of different concentrations of the compound shown in Formula 1 on the viability of NCI-N87 cells was determined by the CCK-8 experiment.

[0046] Figure 6 This illustrates the relationship between HER2 cell viability and the concentration of the compound shown in Formula 1 in Example 2 of this invention.

[0047] Figure 7 Flow cytometry analysis of the apoptosis induced by the compound shown in Formula 1 in Example 2 of this invention in NCI-N87 cells.

[0048] Figure 8 This is a statistical analysis of the apoptosis rate induced by the compound shown in Formula 1 in Example 2 of the present invention in NCI-N87 cells. n =3, compared with the control group, * p <0.05,** p <0.01, *** p <0.001, **** p <0.0001.

[0049] Figure 9 This is a flow cytometry analysis of the cell cycle arrest induced by the compound shown in Formula 1 in Example 2 of the present invention in NCI-N87 cells.

[0050] Figure 10 This is a statistical analysis of the proportion of NCI-N87 cells induced by the compound shown in Formula 1 in Example 2 of the present invention, n =3, compared with the control group, * p <0.05,** p <0.01, *** p <0.001, **** p <0.0001.

[0051] Figure 11 The results of the observation on the inhibition of NCI-N87 cell clone formation by the compound shown in Formula 1 in Example 2 of the present invention.

[0052] Figure 12 The statistical analysis of the inhibition of NCI-N87 cell colony formation rate by the compound shown in Formula 1 in Example 2 of this invention is presented. n =4, compared with the control group, * p <0.05,** p <0.01, *** p <0.001, **** p <0.0001.

[0053] Figure 13This illustrates the growth of subcutaneous xenograft tumor volume after in vivo administration of different dosages of the compound shown in Formula 1 in Example 2 of the present invention. n =5.

[0054] Figure 14 This is a comparison of subcutaneous xenograft volumes at the treatment endpoint after in vivo administration of different doses of the compound shown in Formula 1 in Example 2 of the present invention. n =5.

[0055] Figure 15 This describes the effect of different dosages of the compound of Formula 1 on the survival rate of tumor-bearing nude mice in Example 2 of the present invention. n =5.

[0056] Figure 16 This describes the effect of different dosages of the compound of Formula 1 on the body weight of tumor-bearing nude mice in Example 2 of the present invention. n =5. Detailed Implementation

[0057] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0058] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0059] Example 1: Preparation of the compound shown in Formula 1

[0060] Step 1: Synthesis of Compound 3

[0061] Compound 1 (5.0 g, 41 mmol), compound 2 (20.5 g, 81.9 mmol) and K2CO3 (17 g, 123 mmol) were dissolved in 50 mL of DMF and stirred continuously at 80°C for 16 h. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by flash chromatography to obtain compound 3 as a gray solid (7 g, 58.7%).

[0062] Step 2: Synthesis of Compound 4

[0063] Compound 3 (7.0 g, 24 mmol) was dissolved in 50 ml MeOH, and NaBH4 (1.8 g, 48 mmol) was added. The mixture was stirred continuously at room temperature for 3 h, concentrated under reduced pressure, and purified by flash chromatography to obtain compound 4 as a gray solid (7 g, 99%).

[0064] Step 3: Synthesis of Compound 6

[0065] Compound 4 (5.0 g, 17 mmol) and NaH (3.4 g, 85 mmol) were dissolved in 80 mL THF and stirred at 0 °C for 30 min. Then, compound 5 (10.23 g, 85.28 mmol) was added and stirred at room temperature for 12 h. The reaction was quenched with saturated NH4Cl (50 mL), and extracted with EtOAc (3 × 50 mL). The organic layers were combined, washed with 30 mL of brine, dried over MgSO4, and the solvent was concentrated under reduced pressure. The residue was purified by column chromatography to give compound 6 (1.5 g, 26%) as a gray solid.

[0066] Step 4: Synthesis of the target compound

[0067] A mixture of compound 6 (1.5 g, 4.5 mmol), compound 7 (1.8 g, 9 mmol), Pd(dppf)Cl2 (300 mg, 0.45 mmol), Cs2CO3 (2.94 g, 9 mmol), and dioxane / H2O (20 mL / 5 mL) was added to a three-necked flask, degassed, and purged with nitrogen. The reaction mixture was stirred at 100 °C for 12 h. The solvent was evaporated under reduced pressure, and the residue was further purified by column chromatography to give the target compound (68 mg, 5%) as a white solid.

[0068] Figure 2 The image shows the 1H NMR spectrum of the obtained compound.

[0069] The structural formula of the target compound is as follows Figure 2 The 1H NMR spectrum results, with the chemical formula C 23 H 21 NO 2。 The molecular weight detection results of the target compound are as follows: Figure 3 As shown, the main mass spectrum signal is concentrated at m / z = 366.2, exhibiting C 23 H 21 NNaO2 is a single-charged positive ion [M + Na] + Peak. HPLC detection results of the target compound are as follows: Figure 4 As shown, the purity is greater than 99%.

[0070] Example 2: Biological Activity Study

[0071] This study systematically evaluated the biological effects of a compound on the proliferation, apoptosis, and cell cycle regulation of HER2-positive gastric cancer cells. Based on HER2 expression profiling analysis of gastric cancer cell lines from The Human Protein Atlas database (https: / / www.proteinatlas.org / ), the NCI-N87 cell line (human gastric adenocarcinoma cells) exhibited significant HER2 protein overexpression. Therefore, NCI-N87 cells were selected as a HER2-positive gastric cancer cell model to validate the compound's targeting effect on the HER2 signaling pathway and its antitumor activity.

[0072] The experimental design employed a multi-level functional validation strategy: first, a colony formation assay was used to assess the inhibitory effect of the compound on the long-term proliferation of tumor cells; second, Annexin V-FITC / PI double staining was used to detect changes in cell apoptosis rate; finally, PI single staining cell cycle analysis was combined to elucidate the compound's cell cycle arrest effect on tumor cells, ultimately clarifying the compound's in vivo therapeutic effect on transplanted tumors. Through this systematic study, the aim was to reveal the molecular mechanism by which the candidate compound inhibits the growth of HER2-positive gastric cancer at both the cellular and animal levels, laying an experimental foundation for the further application of the compound.

[0073] Table 1. Experimental Reagents and Consumables

[0074]

[0075] I. Experimental Methods

[0076] (1) Cell clone formation experiment

[0077] 1) Select NCI-N87 cells in logarithmic growth phase, treat the cells with 0.25% trypsin containing EDTA, and then suspend the cells at 6×10⁻⁶. 5 Cells were seeded at a density of 100 μM / dish into Φ35 mm culture dishes. When the cell density approached 70% of the dish, the cells were treated with 100 μM, 300 μM, and 500 μM of the compound shown in Formula 1; the compound shown in Formula 1 was dissolved in DMSO with a stock solution concentration of 10 mg / mL. After culturing the cells for 48 h, the cells were collected and single-cell suspensions were prepared. The cell density was measured using an automated cell counting system. The same volume of DMSO as the 500 μM group was added to the cells as a control group.

[0078] 2) After gently shaking to ensure even distribution of cells, transfer them to a constant temperature incubator for adherent culture and overnight.

[0079] 3) Observe the cell status and clonal growth every 48 hours, and control the total culture period to 18 days.

[0080] 4) After terminating the culture, discard the supernatant, wash the cells twice with PBS, add 1 mL of cell fixation solution, fix at room temperature for 30 min, discard the solution, and air dry for 15 min.

[0081] 5) Add 1 mL of 0.1% crystal violet solution to each dish and immerse at room temperature for 20 min. Then rinse gently with tap water until the background is clear. Allow the culture dishes to air dry at room temperature and count the number of cell clones formed in each group: Cloning rate = number of clones / number of seeded cells × number of cell types.

[0082] (2) Flow cytometry detection of apoptosis

[0083] 1) Select cells in good growth condition, discard the culture medium, wash twice with PBS, and after trypsin digestion, digestion termination, centrifugation, and resuspending, seed the cells into 6-well plates. When the cell density is close to 70% of the culture dish, add drugs (100 μM, 300 μM and 500 μM of the compound shown in Formula 1); grouping is the same as in the cloning experiment.

[0084] 2) Place the 6-well plate in a cell culture incubator and culture for 48 h. Collect the culture medium from each well and slowly rinse the well wall twice with PBS pre-cooled at 4°C. Add 500 μL of 0.25% trypsin without EDTA to digest the cells. Then, stop the digestion with the collected original culture medium to maintain the cell membrane integrity. Finally, transfer the cell suspension to a centrifuge tube.

[0085] 3) Collect cells by centrifugation at 1200 rpm for 3 min, and wash twice with PBS.

[0086] 4) Add 200 μL of pre-prepared 1×binding buffer working solution, resuspend the cells, and then transfer them into a 1.5 mL labeled EP tube.

[0087] 5) Take the experimental group samples and set up 10 μL Annexin V-FITC single staining tubes and 5 μL PI single staining tubes respectively. Add 10 μL Annexin V-FITC and 5 μL PI reagent to each sample of the remaining control group and experimental group respectively.

[0088] 6) After incubating at room temperature in the dark for 15 min, gently mix to form a single-cell suspension, detect by flow cytometry and analyze the results.

[0089] (3) Cell cycle detection

[0090] 1) Preparation of single-cell suspension

[0091] a. Select well-grown cells, discard the culture medium, wash twice with PBS, digest with trypsin, terminate digestion, centrifuge, and resuspend the cells. Seed the cells in Φ35 mm culture dishes. When the cell density is close to 70% of the culture dish, add drugs (100 μM, 300 μM and 500 μM of the compound shown in Formula 1). Group the cells as in the cloning experiment, and collect the cells after 48 h.

[0092] b. Collect the supernatant into a 15 mL centrifuge tube.

[0093] c. Wash once with 1×PBS and collect the cells again into a 15 mL centrifuge tube.

[0094] d. Add 1 mL of EDTA-free trypsin, let stand for 4 min, and observe the cell digestion under a microscope. When the cells become round, add trypsin to a 15 mL centrifuge tube;

[0095] e. Gently tap the bottom of the dish to help cells detach from the surface and form a single-cell suspension;

[0096] f. Add 1×PBS and collect the cells into a 15 mL centrifuge tube;

[0097] g. Centrifuge at 1500 rpm for 5 min and discard the supernatant.

[0098] 2) Periodic staining

[0099] a. Add 500 μL of 1×PBS solution to make a 1×10⁻⁶ solution. 6 The cells were suspended in a 15 mL centrifuge tube.

[0100] b. Slowly add ice-cold anhydrous ethanol to 2 mL, achieving a final concentration of 75%, and store at 4°C overnight.

[0101] c. Centrifuge at 1500 rpm for 5 minutes and discard the supernatant.

[0102] d. Add 1 mL of 1×PBS, suspend the cells, and centrifuge and wash once more.

[0103] e. Discard the supernatant, add 300 μL of PI / RNase staining solution, mix well, and stain at room temperature for 15 min in the dark. Filter the single-cell suspension through a 400-mesh sieve.

[0104] f. On-machine testing.

[0105] (4) Construction of tumor-bearing mouse model and drug treatment

[0106] 1) Construction of tumor-bearing mouse model

[0107] Four-week-old female immunodeficient BALB / c nude mice, after 7 days of acclimatization, were selected and injected subcutaneously into the right axilla using a syringe under aseptic conditions. The cell suspension was adjusted to 5 million cells / 100 μL, with the same volume injected into each mouse. The mice were then housed in an IVC system for approximately 14 days. When the tumor volume reached 100 mm³, they were randomly assigned to groups for subsequent drug treatment.

[0108] 2) Drug treatment

[0109] The experiment included a control group (DMSO diluted with PBS to the same volume as the 5 mg / kg group) and different dose treatment groups (0.25 mg / kg, 2.5 mg / kg, and 5 mg / kg). The drug was administered via tail vein, once every 3 days for 15 days.

[0110] 3) Tumor volume monitoring

[0111] Measure the major diameter (L) and minor diameter (W) of the tumor every 3 days using vernier calipers, and calculate the tumor volume using the following formula:

[0112]

[0113] Where V is the tumor volume (mm³), L is the maximum diameter of the tumor, and W is the minimum diameter of the tumor.

[0114] 4) Data recording and analysis

[0115] During the experiment, changes in the weight and tumor volume of nude mice were recorded. The treatment endpoint was defined as a tumor volume reaching 1500 mm³, at which point the tumor tissue was isolated and photographed. After the experiment, the tumor inhibition effects of each treatment group and the control group were compared using statistical methods, and tumor growth curves were plotted.

[0116] II. Analysis of Experimental Results

[0117] Figure 5 The study showed the inhibitory effect of different drug concentrations on cell proliferation at 24 h and 48 h, with cell viability negatively correlated with drug concentration, exhibiting a dose-dependent effect. At the same concentration, cell viability in the 48 h group was significantly lower than that in the 24 h group, indicating that the inhibitory effect of the drug on cell proliferation increases with prolonged drug treatment time. This time-dependent effect is usually associated with the cumulative effect of the drug or cell cycle arrest. In the low concentration range (0–100 μM), cell viability decreased rapidly, indicating that the drug can significantly inhibit cell proliferation at low doses. In the high concentration range (>100 μM), cell viability tended to stabilize, and cells had essentially stopped proliferating or were approaching a state of death.

[0118] Figure 6The effects of different drug concentrations on cell proliferation after 24 h were demonstrated, and the IC50 value after 24 h of treatment was calculated using nonlinear regression fitting curves. 50 The value was 321 μM. The fitted curve exhibited a classic sigmoid dose-response curve, indicating that the drug's effect conformed to the Hill equation model. From the curve shape, at low concentrations (log value < 4), cell viability remained around 100%, indicating that the drug did not significantly affect cell proliferation at this point. However, as the drug concentration increased, cell viability decreased sharply between Log(Conc.) and cell proliferation, eventually approaching zero, indicating that this range represents the main range of drug action. The experimental data showed a small error margin (standard deviation), indicating good experimental repeatability and data reliability. Furthermore, under high drug concentration conditions (Log(Conc.) > 6), cell viability decreased to extremely low levels, indicating that the drug has strong toxicity to cells at high concentrations.

[0119] Flow cytometry results Figure 7 As shown, the statistical results of apoptosis are as follows: Figure 8 As shown, compared with the control group, the apoptosis rate increased significantly with increasing drug concentration (100 μM, ...). p <0.001; 300 μM and 500 μM, p <0.0001).

[0120] Cell cycle analysis such as Figure 9 , Figure 10 As shown, compared with the control group, 100 μM ( p <0.0001) and 300 μM ( p Treatment with <0.01 significantly increased G0 / G1 phase arrest, while treatment with 500 μM slowed down G0 / G1 phase arrest, but significantly increased G2 / M phase arrest. p <0.001), indicating that the cell cycle arrest effect caused by different treatment concentrations of the compound shown in Formula 1 is different. Low concentration will cause G0 / G1 phase arrest, and as the treatment concentration increases, G2 / M phase arrest gradually increases.

[0121] Cell clonal formation, such as Figure 11 As shown, the number of clonal plaques gradually decreased with increasing drug concentration, and the clonal plaques in the 500 μM group almost completely disappeared. The colony formation rate is as follows: Figure 12 As shown, the colony formation rates of the 100 μM, 300 μM, and 500 μM groups decreased to approximately 35%, 30%, and 10%, respectively, indicating that the drug significantly inhibited cell survival and colony formation. Therefore, the drug can significantly inhibit the survival and colony formation of NCI-N87 cells in a dose-dependent manner.

[0122] like Figure 13 , 14 As shown, compared with the control group, the growth of xenografts in the 0.25 mg / kg, 2.5 mg / kg and 5 mg / kg groups was inhibited, indicating that the compound shown in Formula 1 inhibits the growth of xenografts in a dose-dependent manner in tumor-bearing mice, suggesting that the compound shown in Formula 1 has certain therapeutic potential for HER2-positive gastric cancer.

[0123] like Figure 15 As shown, the survival time of tumor-bearing mice in the 0.25 mg / kg, 2.5 mg / kg and 5 mg / kg groups reached 39 days, 42 days and 57 days, respectively, while the survival rate of tumor-bearing mice in the control group was only 27 days, indicating that the compound shown in Formula 1 can improve the survival rate of tumor-bearing mice.

[0124] like Figure 16 As shown, compared with the initial body weight, the weight of tumor-bearing mice in the 0.25 mg / kg, 2.5 mg / kg and 5 mg / kg groups did not decrease, indicating that the compound shown in Formula 1 has low toxicity to tumor-bearing mice.

[0125] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. The compound shown in Formula 1, 。 2. A method for preparing the compound shown in Formula 1 of claim 1, comprising the following steps: 1) reacting compound 1 and compound 2 under alkaline conditions in an organic solvent to obtain compound 3; 2) Compound 3 undergoes a reduction reaction in the presence of a reducing agent to give compound 4; 3) Compound 4 reacts with bromoacetonitrile (BrCH2CN) to give compound 6; 4) Compound 6 and compound 7 react in the presence of a palladium catalyst and a cesium salt to give the target compound. 。 3. The use of the compound of Formula 1 in claim 1 in the preparation of HER2-positive tumor diagnostic reagents.

4. The application according to claim 3, characterized in that, The application refers to the use of the compound shown in Formula 1 in the preparation of a drug for the prevention and / or treatment of HER2-positive tumors.

5. The application according to claim 4, characterized in that, The tumor is cancer, specifically breast cancer, lung cancer, stomach cancer, or intestinal cancer.

6. The application according to claim 4, characterized in that, In the aforementioned application, the drug has at least one of the following effects: 1) Inhibits the proliferation of tumor cells; 2) Inhibits tumor cell survival and clone formation; 3) Blocking of the tumor cell cycle.

7. A medicament for the prevention and / or treatment of HER2-positive tumors, said medicament comprising the compound shown in Formula 1 of claim 1.

8. The medicament according to claim 7, characterized in that, The tumor is cancer, specifically breast cancer, lung cancer, stomach cancer, or intestinal cancer.

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