Use of dual receptor agonists in treatment of colorectal cancer
By using a combined treatment method of GIP/GLP-1 dual receptor agonists combined with anti-tumor agents, the problem of poor prognosis in some patients with existing colorectal cancer treatments has been solved, and an effective treatment against colorectal cancer cells has been achieved. The improvement of inhibition and metabolic status provides an innovative treatment option for patients with metabolic diseases combined with colorectal cancer.
Patent Information
- Application Number
- CN202411486851.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-23
- Publication Date
- 2025-01-24
AI Technical Summary
Existing treatments for colorectal cancer are poor in some patients and are accompanied by significant side effects, and the biological link between type 2 diabetes and colorectal cancer is unclear, and there is a lack of effective novel therapeutic approaches.
The GIP/GLP-1 dual-receptor agonist is not only used to regulate metabolism, but also has the potential to inhibit the proliferation, migration and invasion of colorectal cancer cells. By combined with the use of antitumor agents such as chemotherapy drugs, immunotherapy agents or anti-virgin agents, To enhance the treatment effect.
Significantly inhibit the proliferation and invasion of colorectal cancer cells, delay or reverse the progress of tumors, improve the metabolic state of patients, and provide an innovative and effective treatment method, especially suitable for colorectal cancer patients with metabolic diseases.
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Figure CN119345327A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the fields of medicine and medicinal chemistry, and in particular to a novel pharmaceutical composition and application thereof in the treatment of colorectal cancer (CRC). Background Art
[0002] Colorectal cancer (CRC) is one of the malignant tumors with the highest morbidity and mortality in the world. Existing treatments include surgery, chemotherapy, and targeted therapy. However, many patients have a poor prognosis with existing therapies, which are accompanied by significant side effects. Therefore, there is an urgent need to explore new and more effective treatment approaches.
[0003] Type 2 diabetes mellitus (T2DM) has been confirmed as an independent risk factor for CRC, and studies have shown that patients with T2DM have a 27% increased risk of CRC. T2DM and CRC share multiple common risk factors, such as age, obesity, smoking, and drinking, but the biological link between the two is still unclear.
[0004] GIP / GLP-1 dual receptor agonists, as a dual receptor agonist of GIP and GLP-1, were initially used to treat T2DM, with significant blood sugar control and weight loss effects, and no significant risk of hypoglycemia. Currently, new therapeutic uses of GIP / GLP-1 dual receptor agonists still need to be developed. Summary of the invention
[0005] The present invention relates to the use of a GIP / GLP-1 dual receptor agonist in the preparation of a drug for treating colorectal cancer, especially in colorectal cancer patients suffering from metabolic diseases (such as type 2 diabetes). The invention reveals that the GIP / GLP-1 dual receptor agonist can not only be used to regulate metabolism, but also has the potential to inhibit the proliferation, migration and invasion of colorectal cancer cells, thereby delaying or reversing the progression of tumors.
[0006] According to the present invention, the GIP / GLP-1 dual receptor agonist has multiple effects, not only inhibiting the malignant progression of tumors, but also improving the blood sugar and blood lipid levels of subjects, and helping to control body weight. This dual receptor agonist is particularly suitable for patients with metabolic diseases (such as type 2 diabetes) and colorectal cancer at the same time, and has significant clinical significance.
[0007] The present invention also provides a method for the combined use of a GIP / GLP-1 dual receptor agonist and an anti-tumor agent, wherein the anti-tumor agent includes but is not limited to a chemotherapeutic drug, an immunotherapy drug, and an anti-angiogenic agent. Through this combination, the therapeutic effect can be further improved, especially when a chemotherapeutic drug such as oxaliplatin, 5-fluorouracil or irinotecan is used in combination with a GIP / GLP-1 dual receptor agonist, the anti-cancer activity can be synergistically enhanced.
[0008] The composition comprises a GIP / GLP-1 dual receptor agonist, a GIP and GLP-1 dual receptor agonist originally used to treat type 2 diabetes (T2DM), which can not only effectively reduce blood sugar and body weight, but also show significant anti-cancer effects. The present invention mainly focuses on the new application of GIP / GLP-1 dual receptor agonist in tumor metabolism and cancer treatment, especially the treatment of colorectal cancer.
[0009] In addition, the present invention also covers a pharmaceutical composition comprising a GIP / GLP-1 dual receptor agonist and at least one chemotherapeutic drug, an immunotherapeutic agent or an anti-angiogenic agent. The composition provides a diverse treatment plan for colorectal cancer, and is particularly suitable for cancer patients who also suffer from metabolic diseases.
[0010] The experimental results show that the GIP / GLP-1 dual receptor agonist can significantly inhibit the proliferation and invasion of colorectal cancer cells, delay tumor progression, and improve the metabolic state of patients. Therefore, the present invention provides an innovative and effective treatment method for colorectal cancer patients, especially those suffering from metabolic diseases.
[0011] The present invention provides use of a GIP / GLP-1 dual receptor agonist in preparing a medicine for treating colorectal cancer.
[0012] In some embodiments, the GIP / GLP-1 dual receptor agonist has a structure as shown in the following Formula I, or a pharmaceutically acceptable salt thereof.
[0013]
[0014] In some embodiments, the subject suffering from the colorectal cancer also suffers from a metabolic disease; preferably, the metabolic disease is type 2 diabetes.
[0015] In some embodiments, the GIP / GLP-1 dual receptor agonist has at least one of the following effects:
[0016] (1) Inhibit the proliferation, migration and / or invasion of colorectal cancer cells,
[0017] (2) Delay and / or reverse tumor progression,
[0018] (3) Improve the blood sugar, blood lipids and / or body weight of the subjects.
[0019] In some embodiments, the GIP / GLP-1 dual receptor agonist is further combined with an anti-tumor agent; preferably, the anti-tumor agent is selected from a chemotherapeutic agent, an immunotherapeutic agent, and / or an anti-angiogenic agent.
[0020] In some embodiments, the chemotherapeutic agent comprises at least one of: the chemotherapeutic agent comprises at least one of the following: oxaliplatin, 5-fluorouracil, irinotecan.
[0021] A pharmaceutical composition comprising a GIP / GLP-1 dual receptor agonist and an anti-tumor agent; preferably, the anti-tumor agent is selected from a chemotherapeutic agent, an immunotherapeutic agent, and / or an anti-angiogenic agent.
[0022] In some embodiments, it includes a GIP / GLP-1 dual receptor agonist, and a chemotherapeutic agent; the chemotherapeutic agent includes at least one of the following: oxaliplatin, 5-fluorouracil, and irinotecan.
[0023] In some embodiments, the GIP / GLP-1 dual receptor agonist is a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt thereof:
[0024] A step comprising contacting colorectal cancer cells with a GIP / GLP-1 dual receptor agonist; preferably, the GIP / GLP-1 dual receptor agonist is a compound with a structure shown in the following formula I or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Figure 1 Schematic diagram of the molecular structure of compound 1.
[0026] Figure 2 To verify the effects of different concentrations of compound 1 on various types of cells. Among them, A is the MC38 cell viability during 4 days of compound 1 treatment, B is the MC38 cell viability after 48 hours of compound 1 treatment, C is the BNL cell viability after 48 hours of compound 1 treatment, D is the U87 cell viability after 48 hours of compound 1 treatment (10 and 50 μM), E is the Pan02 cell viability after 48 hours of compound 1 treatment (10 and 50 μM), F is the HGC cell viability after 48 hours of compound 1 treatment (10 and 50 μM), G is the MKN cell viability after 48 hours of compound 1 treatment (10 and 50 μM).
[0027] Figure 3 Different in vitro experimental methods are used to verify the effect of compound 1 on colon cancer cell MC38. Among them, A is the tumor cell field of view in the transwell chamber, B is the transwell statistical diagram, C is the scratch field of view taken at different times by an inverted microscope, and D is the scratch statistical diagram.
[0028] Figure 4To study the anti-colorectal cancer (CRC) effect of compound 1 in different mouse tumor models. A is the colon cancer bioluminescence imaging of each group of mice, B is the change of bioluminescence radiation efficiency of each group of animals over time, C is the picture of the orthotopic tumor peeled off from each group of animals after killing the mice, D is a representative picture of the subcutaneous tumor peeled off after 21 days of treatment with compound 1 or PBS, E is the statistical graph of the tumor weight of the mice, and F is the statistical graph of the tumor volume of the mice during the experiment.
[0029] Figure 5 The synergistic effect of compound 1 and anticancer drugs in different cells. A is a statistical chart of cell viability detected by CCK-8 method using compound 1 combined with 5-FU, SN-38 or OXA on MC38, and B is a statistical chart of cell viability detected by CCK-8 method using compound 1 combined with 5-FU, SN-38 or OXA on CT26. DETAILED DESCRIPTION
[0030] The technical solution of the present invention is described in detail below with reference to the accompanying drawings. The embodiments of the present invention are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.
[0031] The mouse colon cancer cell lines (MC38 and CT26), human pancreatic adenocarcinoma cell lines (Pan02), human gastric cancer cell lines (HGC27 and MKN45) and mouse embryonic liver cell lines (BNL CL.2) used in this example were purchased from the American Standard Bacteria Library (ATCC, Manassas, Virginia, USA). All cell lines were cultured in DMEM culture medium containing 10% fetal bovine serum (FBS) and antibiotic antifungal agents (Gibco, USA) at 37°C and 5% CO2. All cell lines were regularly tested to ensure that there was no mycoplasma contamination. The culture medium was replaced every other day during the cell culture process until the cell density reached 80% to 90% for cell passage.
[0032] Compound 1, OXA, SN38, and 5-FU were purchased from Selleck (Houston, USA). The above reagents were used for all experiments in this example. The molecule contains two non-coding amino acid residues at the 2nd and 13th amino acids, namely α-aminoisobutyric acid (AIB), and the 20th lysine (K20) is acylated and connected to a γ-glutamic acid (yGlu)-2xOEG linker, which is further connected to a C18 fatty diacid group. This structure enables compound 1 to stimulate GIP and GLP-1 receptors at the same time, and has potential hypoglycemic and weight loss effects.
[0033] Cell Counting Kit-8 (CCK8, Dojindo, Japan) was used to detect the inhibitory effect of compound 1 on the proliferation of various cell lines. MC38, BNL, U87, Pan02, HGC27 and MKN45 cells were seeded in 96-well plates and treated with different doses of compound 1. Cell proliferation was detected at 12, 24, 48, 72 and 96 hours. The absorbance was detected at a wavelength of 450 nm using a microplate reader (BioRad, Berkeley, USA).
[0034] In the combined administration experiment of compound 1 and 5-fluorouracil (5-FU), oxaliplatin (OXA) or SN-38, the concentration of compound 1 was selected as 50 μM, the action concentration of 5-fluorouracil was 5 μM, the action concentration of oxaliplatin was 50 μM, and the action concentration of irinotecan (SN-38) was 100 nM. The CCK8 kit was used to detect the different effects of 5-fluorouracil, oxaliplatin or irinotecan alone and compound 1 with 5-fluorouracil, oxaliplatin or irinotecan on cell proliferation inhibition.
[0035] Cell migration was detected by using Transwell chamber (Corning, 3412). The cells were suspended in serum-free DMEM and added to the upper layer of the chamber, and DMEM containing 10% FBS was added to the lower layer. After 48 hours of migration, the migrated cells were stained with crystal violet, and the images were processed and counted by Adobe Photoshop CC 2018 software.
[0036] MC38 cells were plated in 12-well plates and cultured until the monolayer cells were confluent. Then, scratches were made on the cell monolayer using a 200 μL pipette tip. 10 μM and 50 μM of compound 1 were added to the treatment groups, respectively. The scratches were imaged and analyzed using a DMIL microscope system (Leica, Germany) after 0 and 48 hours, and the scratch healing was used to evaluate the migration ability of the cells. To exclude the effect of cell proliferation on scratch healing, 1 μM mitomycin (Selleck) was added during the experiment.
[0037] The mice used in the present example were C57BL / 6J male mice aged 6 to 8 weeks (Jiangsu Jicui Pharmaceutical Co., Ltd.) and were raised under standard conditions.
[0038] The tumor-bearing mouse model used in this embodiment adopts subcutaneous tumor formation and colon in situ tumor formation. For subcutaneous tumor formation, the specific implementation method is: after the mouse is anesthetized with isoflurane and shaved and cleaned, a 50 μL volume of a cell suspension containing 500,000 MC38 cells (the solvent is PBS) is subcutaneously injected into the right back of the mouse with a microsyringe, and the weight and (colon cancer) tumor volume of the mouse are recorded after 7 days. On the 8th day, each group of mice was given PBS or compound 1 (50nmol / kg) by intraperitoneal injection using a 1mL syringe. The treatment lasted for 21 days, and the tumor volume and mouse weight were measured every 2 days. The tumor volume was measured using a vernier caliper, and the calculation formula was the long diameter of the tumor × the short diameter × the short diameter / 2. The data were plotted and analyzed using GraphPad Prism software, and the statistical method used a two-way analysis of variance. When the mouse tumor volume reached 1000mm 3 The experiment was terminated at the right time. After euthanizing the mice, the subcutaneous tumors of the mice were removed using scissors and forceps, and photos were taken for record.
[0039] For colon orthotopic tumor formation, the specific implementation method is as follows: after the mice were anesthetized with isoflurane, the abdominal hair of the mice was shaved, and the surgical area was disinfected with 75% alcohol. Under sterile conditions, a 50 μL volume of MC38 cell suspension (containing 5×10^5 cells, the solvent was a 1:1 mixture of Matrigel matrix gel and PBS) was injected into the serosal side of the cecal wall using a microsyringe. After injection, the peritoneum and skin of the mice were sutured respectively with sutures. Treatment began on the 35th day after injection. The mice in the experimental group were injected intraperitoneally with 50 nmol / kg of compound 1 every day for 21 days. The mice in the control group were injected with an equal volume of PBS. The growth of the tumor was monitored using the Xenogen IVIS200 bioluminescence imaging system, and the images were analyzed using the LivingImage 2.5 software. At the end of the experiment, the mice were euthanized with isoflurane, and then the cecal orthotopic tumors of the mice were peeled off with scissors and forceps, and photographed and weighed. The tumor samples were used for subsequent omics analysis and molecular detection. Tumor growth data were analyzed using GraphPad Prism software. Statistical analysis methods included two-way analysis of variance (ANOVA), and P values less than 0.05 were considered statistically significant.
[0040] The diabetic model was induced by streptozotocin (STZ) in 6- to 8-week-old C57BL / 6J male mice. STZ was intraperitoneally injected at a dose of 100 mg / kg body weight for 2 days, and blood glucose was measured by a blood glucose meter to verify the hyperglycemic state of diabetic mice.
[0041] Example 1 Compound 1 significantly inhibited the proliferation of colorectal cancer cells in in vitro experiments and was specific to colorectal cancer cells.
[0042] Figure 2 A indicates that compound 1 exhibits a dose-dependent inhibitory effect on the proliferation of colon cancer MC38 cells in the range of 5 μM to 50 μM. Figure 2 B indicates the concentration-dependent inhibitory effect of compound 1 on MC38 at an action time of 48 h. Figure 2 C showed that there was no obvious toxicity to the mouse embryonic liver cell line (BNL CL.2) at this concentration. In order to evaluate the specificity of its anti-tumor effect, the cell proliferation assay was used to detect its effect on four other cancer cell lines. Figure 2 DG showed that compound 1 did not significantly inhibit the proliferation of glioma cells (U87MG), human gastric cancer cells (HGC27 and MKN45), and pancreatic cancer cells (Pan02), indicating that compound 1 is specific for colorectal cancer cells.
[0043] Example 2 Compound 1 significantly inhibited the proliferation of colorectal cancer cells in in vitro experiments, regardless of whether they were in a high-sugar environment.
[0044] An in vitro model of MC38 cells under high glucose (HG) conditions was established in a Transwell system. Figure 3 AB results showed that compound 1 inhibited the migration of cancer cells across the membrane in a dose-dependent manner, and high glucose stimulation had no significant effect on this effect. Figure 3 The CD results showed that in the scratch healing experiment, the cell migration ability of the compound 1-treated group was significantly lower than that of the control group at both concentrations, especially at 50 μM.
[0045] Example 3 In vivo tumor inhibition experiment of compound 1
[0046] In situ tumors and subcutaneous tumors were used to verify the inhibition of tumor growth in tumor-bearing mice by compound 1. In the in situ tumor model, the experiment set up different groups of diabetic and non-diabetic groups to be treated with compound 1 to verify the different therapeutic effects of compound 1, with 10 mice in each group. On the 7th day after the mice formed tumors, equal volumes of PBS and compound 1 (50nmol / kg) were given respectively. In the in situ carcinoma experiment, after the tumor formed on the 7th day, bioluminescence photography was performed every 3 days to record the changes in tumor growth trend. In the subcutaneous tumor experiment, the weight and tumor volume of the mice were recorded after the tumor formed on the 7th day.
[0047] Figure 4 AC showed that compound 1 inhibited orthotopic colorectal cancer growth in both diabetic and non-diabetic mice as evidenced by in vivo imaging and tumor volume measurement. Figure 4 D is a picture of the subcutaneous tumor model mice after tumor removal. Compared with the control group, the tumor volume of the experimental group mice is smaller. Figure 4 E is the mouse tumor mass map, Figure 4 F is the continuous detection result of tumor volume, and the results all show that the volume growth and mass of the tumor are inhibited after the injection of compound 1. This indicates that compound 1 can effectively inhibit the growth of tumors in vivo.
[0048] Example 4 Synergistic effect of compound 1 and anticancer drugs
[0049] Figure 5 A The results showed that the combination of compound 1 and oxaliplatin can enhance the anti-tumor effect of MC38. Figure 5 B. Results show that compound 1 combined with 5-FU or SN-38 can also synergistically delay the growth of CT26. These results indicate that the combination with chemotherapeutic drugs has a synergistic anticancer effect.
Claims
1. Use of a GIP / GLP-1 dual receptor agonist in the preparation of a drug for treating colorectal cancer.
2. The use according to claim 1, characterized in that The agonist is an active ingredient having a structure as shown in the following formula, or a pharmaceutically acceptable salt thereof.
3. The use according to claim 1 or 2, characterized in that The colorectal cancer patients also suffer from metabolic diseases.
4. The use according to claim 3, characterized in that The metabolic disease is type 2 diabetes.
5. The use according to any one of claims 1 to 4, characterized in that: The GIP / GLP-1 dual receptor agonist is further combined with an anti-tumor agent selected from one or more of a chemotherapeutic agent, an immunotherapeutic agent, and an anti-angiogenic agent.
6. The use according to claim 5, characterized in that The chemotherapeutic agent is selected from one or more of oxaliplatin, 5-fluorouracil and irinotecan.
7. A pharmaceutical composition, characterized in that The invention comprises a GIP / GLP-1 dual receptor agonist and an anti-tumor agent, wherein the anti-tumor agent is selected from one or more of a chemotherapeutic agent, an immunotherapeutic agent, and an anti-angiogenic agent.
8. The pharmaceutical composition according to claim 7, characterized in that The chemotherapeutic agent is selected from one or more of oxaliplatin, 5-fluorouracil and irinotecan.
9. The pharmaceutical composition according to claim 7 or 8, wherein The GIP / GLP-1 dual receptor agonist is a compound of the following formula or a pharmaceutically acceptable salt thereof.
10. A method comprising contacting colorectal cancer cells with a GIP / GLP-1 dual receptor agonist, wherein the GIP / GLP-1 dual receptor agonist has a structure as shown in the following formula or a pharmaceutically acceptable salt thereof.