Use of amoxapine in the preparation of a medicament for treating tumors

Abstract

This invention provides the application of amoxapine in the preparation of drugs for treating tumors, belonging to the field of clinical medical technology. Specifically, the tumor refers to a solid tumor that highly expresses MELK. Amoxapine can form hydrogen bonds and CH-π interactions with key amino acid residues at the MELK binding site, thereby achieving a targeted effect on MELK and further inhibiting the proliferation of cancer cells. Simultaneously, by combining virtual screening with multidimensional data analysis, candidate molecules with good binding activity and conformational stability can be rapidly screened, providing a new technical means for the development of MELK inhibitors.

Description

Technical Field

[0001] This invention relates to the field of clinical medical technology, and in particular to the use of amoxapine in the preparation of drugs for treating tumors. Background Technology

[0002] Maternal embryonic leucine zipper kinase (MELK) belongs to the AMPK serine / threonine kinase family and is abnormally highly expressed in various malignant tumors, participating in the regulation of cell cycle, proliferation, apoptosis, and the maintenance of tumor stemness. Studies have shown that MELK expression levels are significantly elevated in various solid tumors such as liver cancer, melanoma, and colon cancer, and are closely associated with poor patient prognosis. Therefore, MELK has become one of the important targets for anti-tumor drug development.

[0003] Currently, some progress has been made in the development of small molecule inhibitors targeting MELK, but most candidate drugs suffer from problems such as low selectivity, significant toxic side effects, or unfavorable pharmacokinetic properties, and no MELK-targeting drugs have yet been approved for marketing. There remains an urgent clinical need for highly effective and low-toxicity MELK inhibitors.

[0004] Amoxapine (molecular formula C) 17 H 16 ClN3O is a traditional antidepressant, belonging to the tetracyclic class of antidepressants, primarily used to treat depression and anxiety disorders. In recent years, drug repositioning strategies have provided new insights into anti-tumor drug development. Studies have explored new uses for traditional drugs in cancer treatment. Furthermore, immune checkpoint inhibitors (such as PD-1 monoclonal antibodies) and anti-microtubule drugs are important current cancer treatments; however, the former has limited response rates, while the latter has significant toxic side effects and is prone to drug resistance. Clinically, there is an urgent need for combination therapy strategies that can sensitize immunotherapy and reduce chemotherapy toxicity. Whether combining MELK inhibitors with the aforementioned drugs has a synergistic effect is currently unknown. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides the application of amoxapine in the preparation of drugs for treating tumors. Through a combination of virtual screening and biological experiments, new uses for known drugs like amoxapine are developed, providing new candidate drugs and combination therapy strategies for tumor treatment. Simultaneously, the drug combination mechanism of amoxapine is explored, offering new drug combination regimens for tumor treatment.

[0006] In order to achieve the objective of this invention, the following technical solution is adopted: This invention provides amoxapine (molecular formula C) 17 H 16Application of ClN3O in the preparation of drugs for treating tumors.

[0007] Furthermore, the tumor is a solid tumor.

[0008] Furthermore, the tumor is a tumor that highly expresses MELK.

[0009] Furthermore, the tumor is any one of liver cancer, colon cancer, or melanoma.

[0010] Furthermore, the amoxapine screening method includes the following steps: S1. Using molecular docking, the small molecules in the compound library are docked and scored at least one binding site of the MELK protein. S2. Select compounds that meet the following criteria: docking score ≤ -7, binding free energy (MM-GBSA) calculated by molecular mechanics / generalized Born surface area method ≤ -35 ​​kcal / mol, and ligand strain energy ≤ 10 kcal / mol. S3. Perform protein-ligand interaction fingerprint analysis on the compounds screened in S2; S4. Perform cluster analysis on the analyzed compounds, set a similarity threshold of ≥85%, and select the compound with the best standard precision pattern in each class as candidate molecules.

[0011] Furthermore, in the protein-ligand interaction fingerprint analysis of S3, the amino acid residues that interact with the ligand at high frequency include at least one of ILE17, CYS89, or GLU93.

[0012] The present invention also provides a pharmaceutical composition comprising: amoxapine, a PD-1 monoclonal antibody, and an antimicrotubule drug.

[0013] Furthermore, the PD1 monoclonal antibody is selected from either nivolumab or pembrolizumab. The antimicrotubule drugs are selected from any one of eribulin, paclitaxel, docetaxel, or vincristine.

[0014] Furthermore, the microtubule-type drug is eribulin.

[0015] Furthermore, the pharmaceutical composition comprises, by weight parts: 1-20 parts amoxapine, 5-15 parts PD1 monoclonal antibody, and 0.5-3 parts antimicrotubule drug.

[0016] Furthermore, the amoxapine is 5-20 parts, the PD1 monoclonal antibody is 10 parts, and the antimicrotubule drug is 1.4 parts.

[0017] The present invention also provides the use of the pharmaceutical composition in the preparation of a medicament for treating tumors.

[0018] Furthermore, the tumor is selected from any one of liver cancer, colon cancer, or melanoma.

[0019] Furthermore, the drug composition is administered in a 21-day dosing cycle; The time interval between the administration of amoxapine and antimicrotubule drugs should be at least 8 hours.

[0020] Furthermore, amoxapine is administered every three days; The PD1 monoclonal antibody is administered on the first day of the dosing cycle; Antimicrotubule drugs are administered on the first and eighth days of the dosing cycle.

[0021] Furthermore, the intraperitoneal injection dose of amoxapine is 5 mg / kg / day to 10 mg / kg / day; the oral dose is 1 mg / kg / day to 5 mg / kg / day. The dosage of PD1 monoclonal antibody is 10 mg / kg; The dosage of antimicrotubule drugs is 1.4 mg / kg.

[0022] The present invention has the following technical effects: (1) Molecular docking results showed that amoxapine forms hydrogen bonds and CH-π interactions with key amino acid residues at the MELK binding site, indicating that amoxapine has good MELK targeting ability.

[0023] (2) Amoxapine can significantly inhibit the proliferation of liver cancer cells such as HepG2 and Hepa1-6, and its inhibitory effect on MELK knockout cells is significantly weakened, which confirms that its anti-tumor effect depends on the MELK target.

[0024] (3) By combining virtual screening with multidimensional data analysis, candidate molecules with good binding activity and conformational stability can be quickly screened out, providing a new technical means for the development of MELK inhibitors.

[0025] (4) By combining drugs, the neurotoxicity and bone marrow suppression risks of antimicrotubule drugs are reduced, while synergistic inhibition of tumor proliferation is achieved, thus improving the antitumor efficacy while ensuring safety. Attached Figure Description

[0026] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0027] Figure 1 Amoxapine molecular structure formula C 17 H 16 ClN3O.

[0028] Figure 2 The results of a virtual screening of 1533 molecules in the compound library L1000; Figure 3 Cell viability was detected using a CCK-8 assay kit. Figure 4 Results of molecular docking analysis; Figure 5 The result of the amoxapine dissociation constant calculated using surface plasmon resonance (SPR) technology; Figure 6 The effect of amoxapine on liver cancer was detected at the animal level. A shows a photograph of the volume change of Hepa1-6 liver cancer; B shows the growth curve of Hepa1-6 liver cancer; and C shows the weight change of the tumor. Figure 7 The effect of amoxapine on colorectal cancer was detected at the animal level. A shows the changes in the volume of MC38 in colorectal cancer; B shows the growth curve of the volume of MC38 in colorectal cancer; and C shows the changes in the weight of the tumor. Figure 8 The effect of amoxapine on melanoma was detected at the animal level; where A is a photograph of the volume change of melanoma B16; B is the growth curve of melanoma B16 volume; C is the weight change of the tumor. Figure 9 The sensitizing effect of amoxapine on PD1 monoclonal antibodies was detected at the animal level. A is a photograph of the volume change of Hepa1-6 liver cancer; B is the growth curve of liver cancer volume; and C is the weight change of the tumor. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0030] In a first aspect, the present invention provides the use of amoxapine in the preparation of medicaments for treating tumors.

[0031] In some embodiments, the tumor is a solid tumor.

[0032] In some embodiments, the tumor is a tumor that highly expresses MELK.

[0033] In some embodiments, the tumor is any one of liver cancer, colon cancer, or melanoma.

[0034] In some embodiments, the amoxapine screening method includes the following steps: S1. Using molecular docking, the small molecules in the compound library are docked and scored at least one binding site of the MELK protein. S2. Select compounds that meet the following criteria: docking score ≤ -7, binding free energy (MM-GBSA) ≤ -35 ​​kcal / mol calculated by molecular mechanics / generalized Born surface area method, and ligand strain energy ≤ 10 kcal / mol. S3. Perform protein-ligand interaction fingerprint analysis on the compounds screened in S2; S4. Perform cluster analysis on the analyzed compounds, set a similarity threshold of ≥85%, and select the compound with the optimal binding free energy obtained by molecular mechanics calculation in each class as the candidate molecule.

[0035] In some embodiments, the amino acid residues that interact frequently with the ligand in the protein-ligand interaction fingerprint analysis in S3 include at least one of ILE17, CYS89, or GLU93.

[0036] In a second aspect, the present invention also provides a pharmaceutical composition comprising: amoxapine, a PD1 monoclonal antibody, and an antimicrotubule drug.

[0037] In some embodiments, the PD1 monoclonal antibody is selected from either nivolumab or pembrolizumab. The antimicrotubule drugs are selected from any one of eribulin, paclitaxel, docetaxel, or vincristine.

[0038] In some embodiments, the microtubule-like drug is eribulin.

[0039] In some embodiments, the pharmaceutical composition comprises, by weight, 1-20 parts amoxapine, 5-15 parts PD1 monoclonal antibody, and 0.5-3 parts antimicrotubule drug.

[0040] In some embodiments, the amoxapine is 5 to 20 parts, the PD1 monoclonal antibody is 10 parts, and the antimicrotubule drug is 1.4 parts.

[0041] Thirdly, the present invention also provides the use of the above-mentioned pharmaceutical composition in the preparation of a medicament for treating tumors.

[0042] In some embodiments, the tumor is selected from any one of liver cancer, colon cancer, or melanoma.

[0043] In some embodiments, the pharmaceutical composition is administered in a 21-day dosing cycle; The time interval between the administration of amoxapine and antimicrotubule drugs should be at least 8 hours.

[0044] In some embodiments, amoxapine is administered every three days; The PD1 monoclonal antibody is administered on the first day of the dosing cycle; Antimicrotubule drugs are administered on the first and eighth days of the dosing cycle.

[0045] In some embodiments, the intraperitoneal injection dose of amoxapine is 5 mg / kg / day to 10 mg / kg / day; the oral dose is 1 mg / kg / day to 5 mg / kg / day. The dosage of PD1 monoclonal antibody is 10 mg / kg; The dosage of antimicrotubule drugs is 1.4 mg / kg.

[0046] The following is a detailed explanation using specific embodiments: Example 1: Screening method for amoxapine 1.1 Virtual Filtering 1.1.1 Preparation of the compound library The compound library selected for this virtual screening was L1000. During the screening process, the compound library was first processed using the Lig Prep module in the Schrödinger software, with the force field set to OPLS4. The Epik method was used to protonate and desalt the compound library at pH 7.0±2.0, generating tautomers while maintaining the original atomic chirality.

[0047] 1.1.2 Protein Preparation The Protein Preparation Wizard module in Schrödinger software was used to perform bond-level optimization, hydrogenation, disulfide bond allocation, hydrogen bond allocation, and energy minimization on the protein. The binding sites for inhibitors were selected in this screening, and the properties of the docking pocket were calculated using the Receptor Grid Generation module in Schrödinger software.

[0048] 1.1.3 Virtual Filtering Import the receptor and ligand files prepared in steps 1.1.1 and 1.1.2, and adopt a step-wise strategy (i.e., SP [Standard Precision Mode] → XP [Ultra-High Precision Mode] → MM-GBSA) to progressively increase the accuracy of the screening. Each step uses a small molecule flexible docking method, and energy optimization is performed after docking. The top 80% of small molecules in the compound library are retained for the next round of screening.

[0049] 1.2 Results Analysis 1.2.1 Selection based on affinity The docking scores of the 1533 compound molecules obtained after screening and deduplication from the L1000 compound library ranged from -14.815 to -4.598, while the MM-GBSA scores ranged from -84.79 kcal / mol to 60.55 kcal / mol. The top 209 molecules with the highest MM-GBSA binding free energy scores were selected, and then the top 30 molecules were chosen from these 209 molecules to examine their inhibitory effect on HepG2 cell proliferation using a CCK-8 assay kit.

[0050] The results showed that drugs 16 (clomipramine hydrochloride) and 23 (amoxaprine, the molecular formula of which is as follows) were effective. Figure 1 (As shown) It showed good inhibitory effect on HepG2 liver cancer cells, Student's t-test. P<0.05, P<0.01, P<0.001.

[0051] The mass, docking score, and MM-GBSA distribution of all screened compound molecules are shown in the table below. Figure 2 Based on the distribution characteristics, the screening criteria were set as docking score ≤ -7 and MM-GBSA value ≤ -35 ​​kcal / mol. After screening, 532 compounds met the criteria for further study.

[0052] 1.2.2 Protein-ligand interaction analysis (PLIF) The results were then imported into MOE to perform receptor-ligand interaction fingerprinting. Protein-ligand Interaction Fingerprint (PLIF) analysis was used to examine the amino acid residues that interact with small molecule compounds. PLIF analysis results showed that 512 compounds from the previous screening step could interact with proteins, and the amino acid residues interacting with small molecules mainly included ILE17, CYS89, and GLU93. Among these, small molecules interacting with CYS89 accounted for the highest proportion of the total number of compounds screened.

[0053] 1.2.3 Ligand Analysis The energy change during the conformational change of a molecule before and after binding to a protein is called ligand strain energy, which is closely related to the conformational stability of the ligand molecule. To exclude compounds with unstable conformations during binding, molecules with ligand strain energy ≤ 10 kcal / mol and ligand efficiency (docking score / number of heavy atoms) ≤ -0.3 were screened for further study. A total of 269 molecules were obtained after screening.

[0054] 1.2.4 Cluster Analysis Cluster analysis was performed using the Cluster module of the MOE software, and the MACCS molecular fingerprint was selected to calculate the similarity between molecules. When the similarity threshold was set to 85%, the results were clustered into 209 categories. For each cluster, the molecule with the best score calculated using MM-GBSA combined with free energy was selected.

[0055] Example 2: Investigation of the efficacy of clomipramine hydrochloride and amoxapine against wild-type Hepa1-6 liver cancer cells and MELK knockout liver cancer cells in mice. Cancer cells were digested and collected, and prepared into a single-cell suspension. Cell counting was performed, and the cells were diluted to the desired density with complete culture medium, 5000 cells per well (100 μL).

[0056] Following the instructions of the CCK-8 assay kit (purchased from Yisheng Biotechnology), cell viability was assessed to investigate the efficacy of clomipramine hydrochloride and amoxapine against mouse wild-type Hepa1-6 hepatocellular carcinoma cells and MELK knockout hepatocellular carcinoma cells. Results are as follows: Figure 3 As shown, amoxapine significantly inhibited the growth of MELK knockout cells compared to clomipramine hydrochloride, according to Student's t-test. P<0.05, P<0.01, P<0.001, P<0.0001.

[0057] Example 3: Combination of MELK and amoxapine First, molecular docking analysis was performed using Discovery Studio software. The analysis showed a docking fraction of -8.406 and a MM-GBSA binding energy of -60.08 kacl / mol (e.g., ...). Figure 4 As shown in the figure, amoxapine forms hydrogen bonds with E93 and E136 of the MELK protein binding site and forms a CH-π interaction with F151.

[0058] Surface plasmon resonance (SPR) technology was applied, with human MELK protein as the stationary phase and amoxapine at concentrations of 25, 12.5, 6.25, 3.12, and 1.56 μM as flow samples. The dissociation time was 60 s, the flow rate was 30 μl / min, and the temperature was 25 ℃. The binding and dissociation signals were monitored in real time, and the in vitro binding affinity of human MELK to amoxapine was calculated. The experimental results are as follows: Figure 5 As shown, the dissociation constant of amoxapine is 4.86 × 10⁻⁶. -5 M indicates that amoxapine has a strong binding affinity to the MELK protein.

[0059] Example 4: Detection of the inhibitory effect of amoxapine on the proliferation of Hepa1-6 liver cancer cells in C57BL / 6 mice tumor-bearing experiments. Hepa1-6 cell line xenograft experiments were conducted on C57BL / 6 mice. Four to six-week-old C57BL / 6 mice were subcutaneously inoculated with Hepa1-6 cells to establish a subcutaneous tumor-bearing model. One week later, amoxapine was administered intraperitoneally at a concentration of 20 mg / kg (purchased from Shanghai Taoshu Technology Co., Ltd.), every three days. Monitoring its effect on the tumor revealed the following experimental results: Figure 6 As shown, amoxapine has a significant inhibitory effect on liver cancer. Figure 6 The tumor volume has decreased as seen in AB. Figure 6 As can be seen in C, the tumor weight decreased. (Student's t-test) P<0.05, P<0.01, P<0.001.

[0060] Amoxapine can significantly inhibit the growth of liver cancer in mice.

[0061] Experiment Example 5: Detection of the inhibitory effect of amoxapine on the proliferation of colon cancer MC38 cells in C57BL / 6 mice tumor-bearing experiments. MC38 cell line xenograft experiments were conducted on C57BL / 6 mice. MC38 cells were subcutaneously inoculated into 4-6 week old C57BL / 6 mice to establish a subcutaneous tumor-bearing model. One week later, amoxapine was administered intraperitoneally at a concentration of 20 mg / kg (purchased from Shanghai Taoshu Technology Co., Ltd.), every 3 days. Monitoring its effect on the tumor revealed the following experimental results: Figure 7 As shown, amoxapine has a significant inhibitory effect on colon cancer. Figure 7 The tumor volume has decreased as seen in AB. Figure 7 As can be seen in C, the tumor weight decreased. (Student's t-test) P<0.05, P<0.01, P<0.001.

[0062] Experiment Example 6: Detection of the inhibitory effect of amoxapine on the proliferation of melanoma B16 cells in C57BL / 6 mice tumor-bearing experiments B16 cell line xenograft experiments were conducted on C57BL / 6 mice. Four to six-week-old C57BL / 6 mice were subcutaneously inoculated with B16 cells to establish a subcutaneous tumor-bearing model. One week later, amoxapine was administered intraperitoneally at a concentration of 20 mg / kg (purchased from Shanghai Taoshu Technology Co., Ltd.), every three days. Monitoring its effect on the tumor revealed the following experimental results: Figure 8 As shown, amoxapine has a significant inhibitory effect on melanoma. Figure 8 The tumor volume has decreased as seen in AB. Figure 8 As can be seen in C, the tumor weight decreased. (Student's t-test) P<0.05, P<0.01, P<0.001.

[0063] Example 7: The antitumor effect of amoxapine in inhibiting tumor growth and sensitizing PD1 therapeutic antibodies. C57BL / 6 mice aged 4-6 weeks were subcutaneously inoculated with Hepa1-6 cells to induce tumor formation, at a dose of 5×10⁶ cells. 6 One week later, mice were administered the drug and divided into four groups. The treatment groups received intraperitoneal injections of amoxapine (20 mg / kg) (purchased from Shanghai Taoshu Technology Co., Ltd.), PD-1 monoclonal antibody (10 mg / kg) (purchased from Celec Chemical Reagent Co., Ltd.), and amoxapine plus PD-1 monoclonal antibody, respectively. Administered the drugs every three days for a total of two doses. Tumor volume and body weight were measured simultaneously. Experimental results are as follows: Figure 9 As shown, from Figure 9 As can be seen from AB, the tumor volume in mice is significantly reduced. Figure 9 As can be seen from C, the tumor mass is reduced; this shows that amoxapine can significantly inhibit the growth of Hepa1-6 cells, and its efficacy is even more significant when combined with PD1.

[0064] Experiment Example 8: Evaluation of the efficacy and safety of three-drug combination therapy for liver cancer 8.1 Laboratory Animals and Models: Animals: C57BL / 6 mice, 4-6 weeks old, female, 8-10 per group; Model: Hepa1-6 cells were subcutaneously seeded at 5 × 10⁶ cells per cell line. 6 / Only; Tumor formation: 1 week after inoculation (tumor volume approximately 50-100 mm) 3 Begin administering the medication.

[0065] Table 1: Experimental Groups Following the experimental groupings described above, the drug was administered in a 21-day cycle, with the dosing regimen as shown in Table 2.

[0066] Table 2: Dosing Regimen Note: All intraperitoneal injections are administered uniformly at 5:00 PM, and tail vein injections are administered at 9:00 AM, ensuring that the dosing interval between amoxapine (afternoon) and antimicrotubule drugs (morning) is ≥8 hours.

[0067] 8.2 Efficacy Evaluation Measure tumor volume (V = major diameter × minor diameter) every 3 days. 2 / 2), at the experimental endpoint (day 21), the tumor was removed, weighed, and the tumor inhibition rate was calculated. The experimental results are shown in Table 3.

[0068] Table 3: Efficacy evaluation results of the three-drug combination therapy for liver cancer The tumor volume and weight in the three-drug combination group were significantly smaller than those in the two-drug combination group; the efficacy of the reduced-dose amoxapine group (10 mg / kg) and the high-dose group (20 mg / kg) was comparable, with no statistical difference.

[0069] Safety performance evaluation 8.3. Safety Performance Evaluation Electrocardiograms were recorded before administration and at 2 hours, 6 hours, and 24 hours after administration. Measure the QT interval and calculate the corrected QTc using the Bazett formula (QTc = QT / ...). ), compare the changes in QTc in each group.

[0070] Table 4: Safety Performance Evaluation By administering amoxapine and eribulin at an interval of ≥8 hours, the risk of QTc prolongation can be significantly reduced; when amoxapine is further reduced to 10 mg / kg, the QTc prolongation completely returns to the level of single-drug safety.

[0071] Neurobehavioral scores were assessed weekly, and the results are shown in Table 5.

[0072] Table 5: Neurobehavioral Scores The addition of amoxapine can antagonize the nerve damage caused by eribulin to a certain extent (high-dose group). In particular, the reduced-dose amoxapine group achieved excellent anti-tumor efficacy while the grip strength and hot plate reaction time of mice were close to normal levels and the incidence of neurotoxicity was significantly reduced.

[0073] Experimental Example 9: The efficacy of three-drug combination therapy against colon cancer and melanoma Animal models: C57BL / 6 mice, inoculated with MC38 colon cancer cells and B16 melanoma cells, respectively; The animal models of colon cancer and the groupings of melanoma are shown below: Blank control, the dosing regimen is the same as in Example 8; Amoxapine monotherapy, with the dosing regimen as described in Example 8; Nivolumab monotherapy, dosage regimen as described in Example 8; For eribulin monotherapy, the dosage regimen is as described in Example 8; Amoxapine + nivolumab + eribulin (high-dose amoxapine), dosing regimen as described in Example 8; Amoxapine + nivolumab + eribulin (reduced dose of amoxapine), the dosing regimen is as described in Example 8.

[0074] The testing indicators and testing content are as described in Example 8.

[0075] Table 6: Efficacy evaluation of the drug in the MC38 colon cancer model (day 21) Table 7: Efficacy evaluation of the drug in the melanoma B16 model (day 21) In MC38 colon cancer and B16 melanoma models, the combination of amoxapine, nivolumab, and eribulin also showed significant synergistic antitumor effects (tumor inhibition rates exceeding 85%). Furthermore, the reduced-dose amoxapine (10 mg / kg) group achieved tumor inhibition effects comparable to the high-dose (20 mg / kg) group in both models, indicating that this reduced-dose combination strategy has universal potential for synergistic enhancement and toxicity reduction in different solid tumor types, and is widely applicable to the treatment of liver cancer, colon cancer, and melanoma.

[0076] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the technical solutions of the embodiments of the present invention.

Claims

1. The application of amoxapine in the preparation of drugs for treating tumors, characterized in that, The tumor is any one of liver cancer, colon cancer, or melanoma.

2. The use of amoxapine according to claim 1 in the preparation of a medicament for treating tumors, characterized in that, The screening method for amoxapine includes the following steps: S1. Using molecular docking, the small molecules in the compound library are docked and scored at least one binding site of the MELK protein. S2. Select compounds that meet the following criteria: docking score ≤ -7, binding free energy ≤ -35 ​​kcal / mol obtained by molecular mechanics calculation, and ligand strain energy ≤ 10 kcal / mol. S3. Perform protein-ligand interaction fingerprint analysis on the compounds screened in S2; S4. Perform cluster analysis on the analyzed compounds, set a similarity threshold of ≥85%, and select the compound with the optimal binding free energy obtained by molecular mechanics calculation in each class as the candidate molecule.

3. The use of amoxapine according to claim 2 in the preparation of a medicament for treating tumors, characterized in that, The amino acid residues that interact frequently with the ligand in the protein-ligand interaction fingerprint analysis in S3 include at least one of ILE17, CYS89, or GLU93.

4. A pharmaceutical composition, characterized in that, The pharmaceutical composition includes amoxapine, a PD-1 monoclonal antibody, and an antimicrotubule drug.

5. The pharmaceutical composition according to claim 4, characterized in that, The PD1 monoclonal antibody is selected from either nivolumab or pembrolizumab. The antimicrotubule drugs are selected from any one of eribulin, paclitaxel, docetaxel, or vincristine.

6. The pharmaceutical composition according to claim 4, characterized in that, The contents, by weight, include: 1-20 parts amoxapine, 5-15 parts PD1 monoclonal antibody, and 0.5-3 parts antimicrotubule drugs.

7. The use of a pharmaceutical composition in the preparation of a medicament for treating tumors, characterized in that, The pharmaceutical composition is the pharmaceutical composition according to any one of claims 4 to 5; The tumor is selected from any one of liver cancer, colon cancer, or melanoma.

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