Application of remegapam in preparation of medicine for treating tumors

Abstract

The invention discloses an application of remegapam in preparation of a medicine for treating tumors. It is found for the first time that remegapam has the effect of remarkably inhibiting tumor growth, the new application of remegapam in the anti-tumor field is disclosed, the indication limitation that the medicine is only used for treating migraine originally is broken through, and the clinical application range of remegapam is expanded.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to the use of Remigipam in preparing a medicine for treating tumors. Background Art

[0002] Calcitonin gene-related peptide (CGRP) is a bioactive neuropeptide composed of 37 amino acids. It is widely distributed in C and Aδ sensory nerve fibers of the central and peripheral nervous systems, and is particularly closely associated with the peripheral vascular network. As an important member of the neuropeptide family, CGRP can be detected in various body fluids and has multiple biological functions, including regulating vascular tone and neuroinflammation. In humans, CGRP exists primarily as two isoforms, α-CGRP and β-CGRP, encoded by the CALCA and CALCB genes, respectively. α-CGRP is primarily expressed in the central and peripheral nervous systems, while β-CGRP is primarily distributed in the brain, intestine, and thyroid tissues. The amino acid sequences of the two proteins share over 90% similarity. The biological effects of CGRP depend on its binding to specific receptors. The functional CGRP receptor is a complex composed of the calcitonin receptor-like receptor (CALCRL), receptor activity-modifying protein 1 (Ramp1), the intracellular protein RCP, and the α-subunit of the Gs protein. Ramp1 is a key accessory protein that determines CGRP binding selectivity and is widely expressed on the membranes of immune cells and various non-immune cells. The complete assembly of this receptor complex is crucial for the transmission of CGRP signals.

[0003] At present, cancer is still one of the major diseases that threaten human health worldwide. Although existing treatment methods include surgery, chemotherapy, radiotherapy, targeted therapy and immunotherapy, these treatment methods all have limitations to varying degrees and cannot meet the demand for high-efficiency, low-toxicity and broad-spectrum treatment strategies under the background of complex tumor pathology. Although immunotherapy drugs represented by monoclonal antibodies have achieved initial success in some tumor types, they also have the following major defects: (1) Complex preparation process and high production cost: their preparation involves multiple technical links such as cell culture, hybridoma construction, and purification, which have extremely high requirements for the production environment and are not conducive to large-scale industrial promotion; (2) The efficacy varies among different populations: the treatment response rate is low in some patients, which is significantly affected by the individual immune background and tumor heterogeneity; (3) The toxic and side effects are more prominent: some monoclonal antibodies lack target specificity and may cause nonspecific attacks on normal cells, inducing immune-related adverse reactions, including rash, endocrine disorders, liver damage, etc., which significantly affect patients' compliance and quality of life.

[0004] Traditional treatments also have numerous shortcomings. For example, surgery has limited effectiveness for advanced or metastatic tumors, resulting in a high rate of residual tumors and long postoperative complications and recovery periods. Chemotherapy lacks targeting, damaging normal tissue while killing tumor cells, leading to serious side effects such as hair loss, bone marrow suppression, and immune deficiency, and can easily induce drug resistance with long-term use. Radiotherapy is limited in its scope by anatomical location and the tolerance threshold of normal tissues, and common local side effects such as radiation-induced skin damage and pneumonia severely impact its efficacy and indication expansion. Therefore, there is an urgent need to develop a small molecule drug with high safety and proven efficacy to overcome the limitations of existing treatment strategies and meet the growing clinical needs of cancer treatment.

[0005] Rimegepant sulfate is a CGRP receptor antagonist marketed for the acute treatment of migraine, with a clear target mechanism, good oral bioavailability, and safety data. In migraine treatment, the drug selectively binds to the CGRP receptor, thereby blocking its binding to endogenous CGRP and inhibiting related signaling pathways. Rimegepant's formulation is mature and well-tolerated, with no severe toxicity observed in long-term use, providing a strong foundation for its translational application in other disease indications. Summary of the Invention

[0006] In view of the defects in the prior art, the present invention proposes the use of Remigipam in the preparation of drugs for treating tumors.

[0007] The present invention provides the use of Remigipam or a pharmaceutical preparation containing Remigipam in preparing a medicine for treating tumors.

[0008] In some embodiments, the pharmaceutical preparation containing remigipam is remigipam sulfate orally disintegrating tablets, remigipam sulfate capsules, or remigipam sulfate oral solution.

[0009] In some embodiments, the tumor is any one of lung cancer, breast cancer, colon cancer, pancreatic cancer, gastric cancer, laryngeal cancer and melanoma.

[0010] In some embodiments, the drug for treating tumors is administered orally.

[0011] In some embodiments, the drug for treating tumors includes a pharmaceutically acceptable carrier and / or a pharmaceutically acceptable carrier.

[0012] In some embodiments, the pharmaceutically acceptable carrier is any one or more of a buffer, an emulsifier, a suspending agent, a stabilizer, a preservative, an excipient, a filler, a coagulant, a conditioning agent, a surfactant, a dispersant, and a defoaming agent.

[0013] In some embodiments, the pharmaceutical carrier is any one or more of a virus, a microcapsule, a liposome, a nanoparticle, and a polymer.

[0014] In some embodiments, the dosage form of the drug includes any one of granules, tablets, pills, and capsules.

[0015] In summary, compared with the prior art, the present invention achieves the following technical effects:

[0016] 1. The present invention discovered for the first time that remegipam has a significant effect in inhibiting tumor growth, revealing its new use in the anti-tumor field; breaking through the original indication limitation of the drug to only treat migraine, and expanding its scope of clinical application.

[0017] 2. Remedipam sulfate orally disintegrating tablets offer advantages such as convenient oral administration, well-defined pharmacokinetic properties, a mature manufacturing process, and high clinical safety, providing a promising translational foundation and industrialization prospects for new use development. Compared to currently used clinical treatments such as monoclonal antibodies and chemotherapy drugs, Remedipam has mild adverse reactions, good tolerability, and is more suitable for long-term maintenance therapy. It can meet the urgent need for highly effective, low-toxic, and highly compliant cancer treatments.

[0018] 3. The new use provided by the present invention further broadens the scope of indications of CGRP receptor antagonists, provides a new small molecule targeted therapy strategy for cancer treatment, and has important medical value and market potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0020] Figure 1 This is the bioinformatics analysis result of Example 1 of the present invention; Figure 1-1 is the chemical structural formula of remegipam sulfate orally disintegrating tablets; Figure 1-2The differential expression of CALCA in 15 tumors and non-tumor normal tissues was analyzed by t-test. The study found that CALCA was highly expressed in 10 tumor tissues, including bladder urothelial carcinoma (BLCA) (P<0.05), colon cancer (COAD) (P<0.0001), renal chromophobe cell carcinoma (KICH) (P<0.01), renal clear cell carcinoma (KIRC) (P<0.0001), renal papillary cell carcinoma (KIRP) (P<0.0001), liver cancer (LIHC) (P<0.05), lung adenocarcinoma (LUAD) (P<0.05), lung squamous cell carcinoma (LUSC) (P<0.01), thyroid cancer (THCA) (P<0.01), and endometrial cancer (UCEC) (P<0.01). Figure 1-3 The study analyzed the differential expression of RAMP1 in 33 tumors and non-tumor normal tissues. The t-test analysis found that RAMP1 was highly expressed in 15 tumor tissues, including bladder urothelial carcinoma (BLCA) (P<0.01), breast cancer (BRCA) (P<0.001), cervical squamous cell carcinoma and adenocarcinoma (CESC) (P<0.01), glioblastoma multiforme (GBM) (P<0.01), head and neck cancer (HNSC) (P<0.05), renal chromophobe cell carcinoma (KICH) (P<0.01), renal clear cell carcinoma (KIRC) (P<0.0001), renal papillary cell carcinoma (KIRP) (P<0.0001), liver cancer (LIHC) (P<0.05), lung adenocarcinoma (LUAD) (P<0.0001), pheochromocytoma and paraganglioma (PCPG) (P<0.01), prostate cancer (PRAD), thyroid cancer (THCA) (P<0.01), and endometrial cancer (UCEC) (P<0.01). Figure 1-4 Figure 3: Effects of high and low expression of CALCA in tumor tissues of lung cancer patients on progression-free survival (PFS), overall survival (OS), and disease-free survival (DFS); (a) and (b) show that low-calca significantly prolonged progression-free survival and overall survival compared with high-calca in tumor tissues of patients with squamous cell lung cancer (P<0.05, P<0.01); (c) shows that low-calca significantly prolonged disease-free survival compared with high-calca in tumor tissues of patients with adenocarcinoma (P<0.05).

[0021] Figure 2 This is the clinical study result of Example 2 of the present invention; Figure 2-1 Elisa was used to measure the protein levels of CGRP and Ramp1 in plasma; Figure 2-2To analyze the difference in RAMP1 protein expression on peripheral blood CD4+ T cells between healthy subjects and lung cancer patients by immunofluorescence; Figure 2-3 To analyze the difference in RAMP1 protein expression on peripheral blood CD4+ T cells between healthy subjects and lung cancer patients by flow cytometry.

[0022] Figure 3 This is a schematic diagram of the changes in tumor volume and weight and the administration method of Lewis lung cancer mice under the intervention of Rimegepant in Experiment 1 of Example 3 of the present invention; Figure 3-1 Schematic diagram of Rimegepant treatment of Lewis lung cancer mice; Figure 3-2 Tumor size of Lewis lung cancer mice after 14 days of treatment; Figure 3-3 This is the growth curve of tumor volume during the treatment of Lewis lung cancer mice; Figure 3-4 Figure 5. Tumor weights of Lewis lung cancer mice after 14 days of treatment.

[0023] Figure 4 This is a schematic diagram of the changes in tumor volume and weight and the administration method of Lewis lung cancer mice under the intervention of Rimegepant in Experiment 2 of Example 3 of the present invention; Figure 4-1 Schematic diagram of Rimegepant treatment of Lewis lung cancer mice; Figure 4-2 Tumor size of Lewis lung cancer mice after 14 days of treatment; Figure 4-3 This is the growth curve of tumor volume during the treatment of Lewis lung cancer mice; Figure 4-4 Figure 5. Tumor weights of Lewis lung cancer mice after 14 days of treatment.

[0024] Figure 5 This is a schematic diagram of the tumor volume, weight, and CGRP protein expression in Lewis lung cancer mice in Experiment 3 of Example 3 of the present invention; Figure 5-1 Schematic diagram of WT, Calca-ko, and Ramp1-ko Lewis lung cancer model mice; Figure 5-2 The tumor size is 10 days after the Lewis lung cancer was established in mice; Figure 5-3 This is the growth curve of tumor volume after Lewis lung cancer mice were formed; Figure 5-4 The tumor weight of Lewis lung cancer mice 10 days after tumor formation; Figure 5-5 The effects of Calca and Ramp1 on CGRP protein in Lewis lung cancer model mice.

[0025] Figure 6 This is a schematic diagram of the changes in tumor volume and weight and the administration method of 4T1 breast cancer mice under the intervention of Rimegepant in Experiment 4 of Example 3 of the present invention; Figure 6-1 Schematic diagram of Rimegepant treatment of 4T1 breast cancer mice; Figure 6-2Tumor size of 4T1 breast cancer mice after 14 days of treatment; Figure 6-3 This is the growth curve of tumor volume during the treatment of 4T1 breast cancer mice; Figure 6-4 Figure 2 shows the tumor weight of 4T1 breast cancer mice after 14 days of treatment.

[0026] Figure 7 This is a schematic diagram of the changes in tumor volume and weight and the administration method of MC38 colon cancer mice under the intervention of Rimegepant in Experiment 5 of Example 3 of the present invention; Figure 7-1 Schematic diagram of Rimegepant treatment of MC38 colon cancer mice; Figure 7-2 Tumor size of MC38 colon cancer mice after 7 days of treatment; Figure 7-3 This is the growth curve of tumor volume during the treatment of MC38 colon cancer mice; Figure 7-4 The weight of the tumor was 7 days after the formation of MC38 colon cancer in mice. DETAILED DESCRIPTION

[0027] To help those skilled in the art better understand the present invention, the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work should fall within the scope of protection of the present invention.

[0028] Rimegepant sulfate orally disintegrating tablets (Rimegepant) is a chemical drug, and its chemical name is (5S, 6S, 9R)-5-amino-6-(2,3-difluorophenyl)-6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl-4-(2-oxo-2,3-dihydro-1H-imidazo[4,5-b]pyridin-1-yl)-1-piperidine-1-carboxylic acid hemisulfate sesquihydrate, and its structural formula is as follows: Figure 1 As shown in 1-1. This application, through in-depth analysis of the Cancer Genome Atlas (TCGA) database, revealed that calcitonin gene-related peptide (CGRP) and its receptor component, receptor activity-modifying protein 1 (Ramp1), are highly expressed in various tumor tissues. Further statistical analysis showed that this high expression is significantly negatively correlated with poor prognosis and lower survival rate in cancer patients.

[0029] To further verify the above findings, this application collected clinical samples for testing and analysis. The results showed that compared with healthy people, the levels of CGRP and Ramp1 proteins in the peripheral blood of lung cancer patients were significantly increased; at the same time, the expression level of Ramp1 in the immune cells of lung cancer patients was also significantly upregulated. Based on the results of clinical studies, this study further conducted animal experiments. In the Lewis lung cancer mouse model, the results showed that the expression of CGRP and Ramp1 genes can significantly promote tumor growth. In-depth research found that this phenomenon is closely related to the role of CGRP-Ramp1 in the immunosuppressive process. In addition, the results of pharmacodynamic experiments showed that after treatment with the CGRP receptor antagonist remdesivir sulfate, tumor growth in the Lewis lung cancer mouse model was significantly inhibited, and tumor volume and weight were significantly reduced. This result indicates that the CGRP-Ramp1 signaling pathway is a potential target for tumor treatment. This study innovatively proposed the application value of remdesivir sulfate in tumor treatment.

[0030] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. The materials and reagents used are all commercially available unless otherwise specified.

[0031] Example 1 Bioinformatics Analysis

[0032] To further explore the immunological characteristics and prognostic evaluation potential of calcitonin gene-related peptide (CGRP) in human tumors, this study conducted a comprehensive pan-cancer analysis. The research data mainly came from the Cancer Genome Atlas (TCGA) database. After obtaining the data, it was strictly standardized to ensure the accuracy and consistency of the data. Subsequently, the R language and professional statistical software were used to analyze the expression levels of CALCA and RAMP1 RNA in different tumor tissues. Figure 1 As shown in 1-2 and 1-3.

[0033] The analysis results showed that there were significant differences in the expression of CALCA and RAMP1 in tumor tissues and non-tumor normal tissues. In order to further clarify its association with tumor prognosis, this example obtained relevant data from the TCGA database and the Gene Expression Omnibus (GEO) database, and conducted survival association analysis on the data of overall survival (OS), progression-free survival (PFS) and disease-free survival (DFS) for tumor types such as lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). The Cox proportional hazard model analysis found that the expression of CALCA was negatively correlated with the progression-free survival, overall survival, and disease-free survival of lung cancer patients, indicating that CALCA and RAMP1 are high-risk genes for lung cancer, such as Figure 1 shown in 1-4.

[0034] Example 2 Clinical Study

[0035] Experimental samples: Fresh blood samples were obtained from lung cancer patients and healthcare workers in the Oncology Department of the Shenzhen Hospital, Cancer Hospital of the Chinese Academy of Medical Sciences.

[0036] Experimental groups: healthy group (Health), non-small cell lung cancer group (NSCLC), small cell lung cancer group (SCLC).

[0037] Experimental steps: Take fresh blood and add it to a tube containing anticoagulant, invert and mix thoroughly, and centrifuge at 1000-2000×g for 10 minutes. The upper layer of light yellow transparent liquid is plasma, which is placed on ice for testing or frozen at -80℃ for later use.

[0038] Evaluation: Plasma CGRP and Ramp1 protein levels were measured using ELISA. Human calcitonin gene-related peptide (CGRP) enzyme-linked immunosorbent assay kit (Human CGRP ELISA kit, Cusabio, Cat#: CSB-E08210h); human receptor activity-modifying protein 1 (Human Ramp1 ELISA Kit, Shanghai Ai Meng You Ning Biotechnology Co., Ltd., Cat#: LV11554). Ramp1 protein expression in peripheral blood was determined by immunofluorescence.

[0039] The results are as follows Figure 2 As shown, ELISA results showed that the CGRP level in the plasma of patients with small cell lung cancer was significantly higher than that in the healthy control group and the non-small cell lung cancer group (p < 0.001, p < 0.05), with no statistical difference between the healthy control group and the non-small cell lung cancer group (p > 0.05). The Ramp1 level in the plasma of patients with small cell lung cancer was significantly higher than that in the healthy control group and the non-small cell lung cancer group (p < 0.0001, p < 0.001), with no statistical difference between the small cell lung cancer group and the non-small cell lung cancer group (p > 0.05). Immunofluorescence assay results showed that the expression of Ramp1 on PBMCs in 1 mL of peripheral blood from patients with non-small cell lung cancer was significantly higher than that in healthy controls.

[0040] Example 3: Pharmacological Efficacy Verification of Remigipam Sulfate Orally Disintegrating Tablets

[0041] Rimegepant sulfate orally disintegrating tablets (Rimegepant) are administered orally. Rigorous experimental research and analysis have determined that the effective dose range for rimegepant in treating tumors is 0.2-0.5 mg / g. Within this dose range, the expected pharmacological effect is achieved. Furthermore, the safe dose range is less than 2.5 mg / g. This dose limit effectively ensures drug safety and minimizes the risk of adverse reactions. This provides an important dosage reference for subsequent tumor-related research and potential clinical applications based on rimegepant.

[0042] (1) Experiment 1

[0043] The effects of different doses of Rimegepant (0.2 mg / g, 0.4 mg / g) on ​​tumor growth in lung cancer-bearing mice were studied. The effect of Rimegepant in inhibiting tumor growth and the dose-effect relationship were analyzed by measuring the mouse body weight and tumor volume daily and measuring the tumor weight after tumor removal.

[0044] Experimental animals: Nine C57BL / 6N females, aged 8 to 12 weeks, weighing 25 g ± 5 g, SPF grade, were selected and housed at the Experimental Animal Center of Southern University of Science and Technology in a constant humidity and temperature barrier environment.

[0045] Experimental groups: PBS group, Rimegepant 0.2 mg / g group, and Rimegepant 0.4 mg / g group, with 3 rats in each group.

[0046] Modeling: Lewis lung cancer cells were thawed and cultured to the logarithmic growth phase to prepare single cell suspension (5×10 5 Lung cancer tumor-bearing mouse models were established by subcutaneously injecting a cell suspension (25 μL PBS + 25 μL Matrigel) into the right groin of model group mice under sterile conditions. The PBS group received an injection of the same dose of PBS solution at the same site. Successful modeling was considered successful when a subcutaneous volume >100 mm³ was observed 5 days after inoculation.

[0047] Administration: After successful modeling, the dosage was calculated based on the standard of 25 g body weight per mouse. Rimegepant drug suspension was prepared with normal saline and administered once daily by gavage, 0.2 mL each time, for 14 consecutive days.

[0048] Evaluation: The body condition, activity, fur, body shape, and mental state of the mice were observed daily. The body weight and tumor volume (mm 3 ), the tumor weight (Tumor weight, g) was measured after the tumor was removed, and a curve was drawn for statistical analysis. The results are shown in Figure 3 shown.

[0049] Results: Comparison of tumor weights after tumor removal revealed that the PBS group > the Rimegepant 0.2 mg / g group > the Rimegepant 0.4 mg / g group (p < 0.01, p < 0.001). The tumor volume growth curve showed that starting from day 5, the tumor volume in the PBS group increased rapidly, while the tumor volume in the Rimegepant 0.2 mg / g and Rimegepant 0.4 mg / g groups increased more slowly than that in the PBS group, namely, the PBS group > the Rimegepant 0.2 mg / g group > the Rimegepant 0.4 mg / g group.

[0050] (2) Experiment 2

[0051] The effects of different doses of Rimegepant (0.25 mg / g group, 0.5 mg / g group) on tumor growth in lung cancer-bearing mice were investigated. By monitoring the changes in mouse body weight, tumor volume and weight, the differences in the inhibitory effects of Rimegepant on tumor growth at different doses were clarified, providing an experimental basis for the optimization of drug dosage.

[0052] Experimental animals: Animal experiments were purchased from Beijing Weitonglihua, C57 / 6N, female, 8-12 weeks old, 9 animals, 3 animals in each group, weighing 25g±5g, SPF grade, and housed in a constant humidity and temperature barrier environment at the Experimental Animal Center of Southern University of Science and Technology.

[0053] Experimental groups: PBS group, Rimegepant 0.25 mg / g group, and Rimegepant 0.5 mg / g group, with 3 rats in each group.

[0054] Modeling: Lewis lung cancer cells were thawed and cultured to the logarithmic growth phase to prepare single cell suspension (5×10 5 Lung cancer tumor-bearing mouse models were established by subcutaneously injecting a cell suspension (25 μL PBS + 25 μL Matrigel) into the right groin of the model group mice under sterile conditions. The PBS group was injected with the same dose of PBS solution at the same site. Five days after inoculation, subcutaneous tumors with a volume of >100 mm 3 The modeling can be considered successful.

[0055] Administration: After successful modeling, the dosage was calculated based on the standard of 25 g body weight per mouse. Rimegepant drug suspension was prepared with normal saline and administered once daily by gavage, 0.2 mL each time, for 14 consecutive days.

[0056] Evaluation: The body condition, activity, fur, body shape, mental state, etc. of the mice were observed daily. At the same time, the body weight and tumor volume (mm 3 ), the tumor weight (Tumor weight, g) was measured after the tumor was removed, and a curve was drawn for statistical analysis. The results are shown in Figure 4 shown.

[0057] Results: Tumor weight comparison after tumor removal revealed that the PBS group > the Rimegepant 0.25 mg / g group > the Rimegepant 0.5 mg / g group (p < 0.01, p < 0.0001). Tumor volume growth curves revealed that tumor volume in the PBS group increased rapidly starting on day 4. Tumor volume in the Rimegepant 0.25 mg / g and Rimegepant 0.5 mg / g groups increased more slowly than that in the WT group, with the PBS group > the Rimegepant 0.25 mg / g group > the Rimegepant 0.5 mg / g group.

[0058] (3) Experiment 3

[0059] Experimental Animals: Nine female C57BL / 6N wild-type (WT) mice, CALCA knockout (KO) mice, and Ramp1 knockout (KO) mice, aged 8-12 weeks and weighing 180-200 g, were selected. They were housed in a SPF-grade laboratory animal center at the Southern University of Science and Technology in a constant humidity and temperature barrier environment.

[0060] Experimental groups: WT group, CALCA-KO group, and Ramp1-KO group.

[0061] Modeling: Lewis lung cancer cells were thawed and cultured to the logarithmic growth phase to prepare single cell suspension (5×10 5 Under sterile conditions, the cell suspension (25 μL PBS + 25 μL Matrigel) was subcutaneously injected into the right groin of the model group mice to establish a lung cancer tumor-bearing mouse model. The PBS group was injected with the same dose of PBS solution at the same site. Five days after inoculation, if a subcutaneous tumor with a volume greater than 100 mm 3 If the tumor is not found, the model is considered to be successful.

[0062] Evaluation: After successful modeling, the mice were observed daily for 10 consecutive days for physical condition, activity, fur, body shape, mental state, etc. The weight and tumor size of the mice were measured, and the expression of CGRP protein in tumor tissue was analyzed by western-blot, and statistical analysis was performed, such as Figure 5 shown.

[0063] Results: Comparison of tumor size revealed that the WT group > CALCA-KO group > Ramp1-KO group (p < 0.001, p < 0.0001). Tumor volume growth curves revealed that starting on day 4, tumor volume in the WT group increased rapidly, while that in the CALCA-KO and Ramp1-KO groups increased more slowly than in the WT group. Western blot analysis revealed that CGRP protein expression was higher in the WT group than in the CALCA-KO group and Ramp1-KO group (p < 0.001, p < 0.001). These results suggest that ablation of CALCA and Ramp1 gene expression can inhibit tumor growth in Lewis lung cancer mice.

[0064] (4) Experiment 4

[0065] Objective: To study the effects of different doses of rimegepant (0.4 mg / g) on ​​tumor growth in breast cancer-bearing mice. The inhibitory effect of rimegepant on tumor growth and the dose-effect relationship were analyzed by measuring the body weight and tumor volume of mice daily and measuring the tumor weight after tumor removal.

[0066] Experimental animals: 9 female BALB / c animals, 8-12 weeks old, weighing 25 g ± 5 g, SPF grade, were selected and housed in a constant humidity and temperature barrier environment at the Experimental Animal Center of Southern University of Science and Technology.

[0067] Experimental groups: PBS group and Rimegepant 0.4 mg / g group, 3 rats in each group.

[0068] Modeling: 4T1 breast cancer cell line was thawed and cultured to the logarithmic growth phase to prepare single cell suspension (5×10 5 Lung cancer tumor-bearing mouse models were established by subcutaneously injecting a cell suspension (25 μL PBS + 25 μL Matrigel) into the right groin of the model group mice under sterile conditions. The PBS group was injected with the same dose of PBS solution at the same site. Five days after inoculation, subcutaneous tumors with a volume of >100 mm 3 The modeling can be considered successful.

[0069] Administration: After successful modeling, the dosage was calculated based on the standard of 25 g body weight per mouse. Rimegepant drug suspension was prepared with normal saline and administered once daily, 0.2 mL each time by gavage, for 7 consecutive days.

[0070] Assessment: Gross observation: Observe the body condition, activity, fur, body shape, mental state, etc. of mice daily. Measure the weight and tumor volume (mm 3), and the tumor weight (g) was measured after the tumor was removed, and a curve was drawn for statistical analysis (eg Figure 6 ).

[0071] Results: After tumor removal, tumor weight was compared, showing that the PBS group was greater than the Rimegepant 0.4 mg / g group (p < 0.05). The tumor volume growth curve showed that starting from day 2, the tumor volume in the PBS group increased rapidly, while the tumor volume in the Rimegepant 0.4 mg / g group increased more slowly than that in the PBS group, i.e., the PBS group was greater than the Rimegepant 0.4 mg / g group (p < 0.001).

[0072] (5) Experiment 5

[0073] Objective: To study the effects of different doses of rimegepant (0.4 mg / g) on ​​tumor growth in MC38 colon cancer-bearing mice. The inhibitory effect of rimegepant on tumor growth and the dose-effect relationship were analyzed by measuring the mouse body weight and tumor volume daily and measuring the tumor weight after tumor removal.

[0074] Experimental Animals: Nine female C57BL / 6N mice, aged 8–12 weeks and weighing 25 g ± 5 g, were selected. They were maintained at the Experimental Animal Center of the Southern University of Science and Technology in a constant humidity and temperature barrier environment.

[0075] Experimental groups: PBS group and Rimegepant 0.4 mg / g group, 3 rats in each group.

[0076] Modeling: MC38 colon cancer cell lines were thawed and cultured to the logarithmic growth phase to prepare single cell suspension (5×10 5 Under sterile conditions, the cell suspension (25 μL PBS + 25 μL Matrigel) was subcutaneously injected into the right groin area of ​​the model group mice to establish a lung cancer tumor-bearing mouse model. The PBS group was injected with the same dose of PBS solution at the same site. Five days after inoculation, if a subcutaneous tumor with a volume greater than 100 mm 3 The modeling can be considered successful.

[0077] Administration: After successful modeling, the dosage was calculated based on the standard of 25 g body weight per mouse. Rimegepant drug suspension was prepared with normal saline and administered once daily by gavage, 0.2 mL each time, for 14 consecutive days.

[0078] Evaluation: Daily observation of the mice's physical condition, activity, fur, body shape, mental state, etc. Daily measurement of mouse body weight, tumor volume (mm 3), and the tumor weight (g) was measured after the tumor was removed, and a curve was drawn for statistical analysis (eg Figure 7 ).

[0079] Results: Tumor weight after tumor removal was compared, showing that the PBS group was greater than the Rimegepant 0.4 mg / g group (p < 0.05). Tumor volume growth curves revealed that starting on day 5, tumor volume in the PBS group rapidly increased, while that in the Rimegepant 0.4 mg / g group grew more slowly than in the WT group, with the PBS group being greater than the Rimegepant 0.4 mg / g group (p < 0.001).

[0080] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Use of Remigipam or a pharmaceutical preparation containing Remigipam in the preparation of a drug for treating tumors.

2. The use according to claim 1, characterized in that The pharmaceutical preparation containing remigipam is remigipam sulfate orally disintegrating tablets, remigipam sulfate capsules or remigipam sulfate oral solution.

3. The use according to claim 1, characterized in that The tumor is any one of lung cancer, breast cancer, colon cancer, pancreatic cancer, gastric cancer, laryngeal cancer and melanoma.

4. The use according to claim 1, characterized in that The drug for treating tumors is administered orally.

5. The use according to claim 1, characterized in that The drug for treating tumors includes a pharmaceutically acceptable carrier and / or a pharmaceutically acceptable carrier.

6. The use according to claim 5, characterized in that The pharmaceutically acceptable carrier is any one or more of a buffer, an emulsifier, a suspending agent, a stabilizer, a preservative, an excipient, a filler, a coagulant, a conditioning agent, a surfactant, a dispersant and a defoaming agent.

7. The use according to claim 5, characterized in that The pharmaceutical carrier is any one or more of viruses, microcapsules, liposomes, nanoparticles and polymers.

8. The use according to claim 1, characterized in that The dosage form of the drug includes any one of granules, tablets, pills and capsules.

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