A pharmaceutical composition for treating tumors, a medicine and application
By combining the anti-angiogenic drug apatinib with the HIF-1a inhibitor CAY10585, the problem of tumor cell hypoxia tolerance caused by anti-angiogenic therapy was solved, and effective inhibition of tumor growth and metastasis was achieved.
Patent Information
- Application Number
- CN202310923757.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-07-26
- Publication Date
- 2025-10-24
- Estimated Expiration
- 2043-07-26
AI Technical Summary
Existing anti-angiogenic therapies, when used to treat tumors, cause hypoxia within the tumor, enhancing the hypoxia tolerance of tumor cells, leading to treatment resistance, and making it difficult to effectively inhibit tumor growth and metastasis.
Combining the anti-angiogenic drug apatinib with the HIF-1a inhibitor CAY10585 weakens the hypoxia tolerance of tumor cells by inhibiting tumor angiogenesis and blocking the glucose metabolism reprogramming of tumor cells.
It enhanced the anti-tumor effect, effectively inhibited tumor growth and metastasis, overcame resistance to anti-angiogenic therapy, and promoted tumor progression.
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Figure CN116850295B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of combination drug, and particularly relates to a pharmaceutical composition for treating tumors, a drug and application. BACKGROUND
[0002] Apatinib is a new type of small molecule tyrosine kinase inhibitor (TKI) independently developed in China, which targets vascular endothelial growth factor receptor 2 (VEGFR-2) and can effectively inhibit tumor angiogenesis, inhibit tumor progression and effectively prolong the progression-free survival and overall survival of patients with advanced tumors and metastatic tumors. Apatinib is the first anti-angiogenic agent approved by the China Food and Drug Administration for the treatment of metastatic gastric cancer. At present, apatinib has been recommended as the third-line treatment for patients with advanced gastric cancer and metastatic gastric cancer, and effectively improves the survival prognosis of tumor patients by inhibiting tumor angiogenesis. However, anti-angiogenic therapy leads to reduced blood supply in the tumor, exacerbating the hypoxia in the tumor, and a selective subset of tumor cells enhances the hypoxia tolerance of cells through glucose metabolism reprogramming, leading to failure of anti-angiogenic therapy. SUMMARY
[0003] The purpose of the present application is to provide a pharmaceutical composition for treating tumors, a drug and application, which can overcome the problem of resistance to anti-angiogenic therapy in clinical practice, weaken the hypoxia tolerance of tumor cells while inhibiting angiogenesis, and enhance the anti-tumor effect.
[0004] The present application provides a pharmaceutical composition for treating tumors, comprising an anti-angiogenic drug and a HIF-1a inhibitor.
[0005] Preferably, the anti-angiogenic drug comprises at least one of apatinib, bevacizumab and ramucirumab.
[0006] Preferably, the HIF-1a inhibitor comprises at least one of CAY10585 and Lificiguat (YC-1).
[0007] Preferably, the working concentration ratio of apatinib and CAY10585 is 120mg / kg:10mg / kg.
[0008] The present application also provides the use of the above-mentioned pharmaceutical composition in the preparation of a drug for treating tumors.
[0009] Preferably, the treatment comprises at least one of the following:
[0010] (1) inhibiting tumor angiogenesis;
[0011] (2) promote the formation of hypoxic microenvironment;
[0012] (3) inhibit the growth of tumor;
[0013] (4) inhibit the metastasis of tumor.
[0014] The application also provides a medicine for treating tumor, the active ingredient of which comprises the above-mentioned medicine composition and pharmaceutically acceptable adjuvant.
[0015] Preferably, the mass percentage of the active ingredient is 0.1-99%.
[0016] Preferably, the dosage form of the medicine comprises oral dosage, intravenous injection or intraperitoneal injection.
[0017] Preferably, the tumor comprises gastric cancer.
[0018] Beneficial effects: the application provides a medicine composition for treating tumor, which combines anti-angiogenic medicine and HIF-1a inhibitor. The embodiment of the application proves that the hypoxic microenvironment induced by anti-angiogenic therapy promotes the accumulation of HIF-1a, the higher the degree of hypoxia in tumor is, the higher the expression of HIF-1a is, and HIF-1a improves the hypoxic tolerance of cells by enhancing glucose metabolic reprogramming, so that tumor cells maintain viability in the hypoxic microenvironment.
[0019] The embodiment of the application also proves that the anti-angiogenic medicine apatinib and HIF-1a inhibitor CAY10585 can produce synergistic complementary effect through combined therapy: apatinib inhibits the formation of tumor neovascularization, the hypoxic microenvironment induced by apatinib promotes the expression of HIF-a, and enhances the targeting of HIF-a inhibitor CAY10585; CAY10585 blocks the glucose metabolic reprogramming of gastric cancer cells and weakens the hypoxic tolerance of tumor cells. The combined therapy of apatinib and CAY10585 helps to overcome the problem of clinical resistance to anti-angiogenic therapy, and enhances the anti-tumor effect by inhibiting angiogenesis and weakening the hypoxic tolerance of tumor cells. The application also proves through in vivo experiments on mice that apatinib effectively inhibits the formation of tumor neovascularization, reduces tumor blood supply and promotes the formation of hypoxic microenvironment. The combined therapy of apatinib and CAY10585 effectively inhibits the growth and metastasis of tumor, enhances the anti-tumor effect and inhibits the progression of tumor. BRIEF DESCRIPTION OF DRAWINGS
[0020] Figure 1 Flow chart for in vivo treatment experiment of the application;
[0021] Figure 2 Result chart of the correlation between hypoxic microenvironment induced by anti-angiogenic therapy and HIF-1a;
[0022] Figure 3 Figure for the correlation of HIF-1a and the reprogramming of glucose metabolism and the hypoxia tolerance of cells;
[0023] Figure 4 Figure for the correlation of HIF-1a inhibitor CAY10585 and the inhibition of the reprogramming of glucose metabolism and the hypoxia tolerance of gastric cancer cells;
[0024] Figure 5 Figure for the effect of the combination therapy of apatinib and CAY10585 and the anti-tumor effect. DETAILED DESCRIPTION
[0025] The present application provides a pharmaceutical composition for treating tumors, comprising an anti-angiogenic drug and a HIF-1a inhibitor.
[0026] The anti-angiogenic drug of the present application preferably comprises at least one of the following: apatinib, bevacizumab and ramucirumab; and the HIF-1a inhibitor comprises at least one of the following: CAY10585 and Lificiguat (YC-1); in the examples, the combination of apatinib and CAY10585 is used as an example for illustration, and the anti-angiogenic drug apatinib (oral gavage, 120 mg / kg, daily) is administered on the 10th day when the gastric cancer orthotopic tumor model is successfully constructed, and CAY10585 (oral gavage, 10 mg / kg, daily) is administered on the 24th day. The present application does not have special limitations on the source of the apatinib and CAY10585, and commercially available products in the art can be used.
[0027] The present application also provides the use of the above-mentioned pharmaceutical composition in the preparation of a medicament for treating tumors.
[0028] The CAY10585 of the present application is an effective hypoxia-inducible factor HIF-1a inhibitor, which can inhibit the accumulation of HIF-1a in cells under hypoxic conditions. As a key transcription factor for sensing changes in external oxygen, HIF-1a is responsible for activating the transcription of genes related to hypoxia homeostasis. This study found that HIF-1a is highly enriched in the hypoxic microenvironment of gastric cancer, and HIF-1a improves the hypoxia tolerance of gastric cancer cells by regulating the reprogramming of glucose metabolism in gastric cancer cells, so that gastric cancer cells maintain viability in a hypoxic microenvironment, escape apoptosis or death. Further leading to resistance to anti-angiogenic therapy of gastric cancer. Based on this molecular mechanism, the combination of apatinib and CAY10585 is used for combined therapy, which inhibits tumor angiogenesis and blocks the reprogramming of glucose metabolism in tumor cells, overcomes the resistance to anti-angiogenic therapy, and inhibits tumor progression.
[0029] The treatment according to the present application preferably comprises at least one of the following:
[0030] (1) inhibiting the formation of new blood vessels in the tumor;
[0031] (2) promoting the formation of a hypoxic microenvironment;
[0032] (3) inhibiting the growth of the tumor;
[0033] (4) inhibiting the metastasis of the tumor.
[0034] The present application also provides a medicament for treating tumors, the active ingredient of which comprises the above-mentioned pharmaceutical composition, and further comprises a pharmaceutically acceptable excipient.
[0035] The mass percentage of the active ingredient according to the present application is preferably 0.1-99%. The dosage form of the medicament according to the present application preferably comprises oral dosage, intravenous injection or intraperitoneal injection.
[0036] In order to further illustrate the present application, the pharmaceutical composition for treating tumors, medicament and application provided by the present application are described in detail below in conjunction with examples, but they should not be understood as limiting the scope of protection of the present application.
[0037] Example 1
[0038] I. Experimental methods
[0039] 1. In vitro treatment experiment:
[0040] Material source: HIF-1a inhibitor CAY10585 was purchased from Selleck Company (product number S8441) Experimental steps:
[0041] (1) Cell culture: adjust the cells to a good state, when the cells are in the logarithmic growth phase, use 0.25% EDTA trypsin to digest the cells, and inoculate them into 24-well plates, and place them in a 37°C cell incubator containing 5% CO2 for further culture; after the cells adhere, replace the complete culture medium containing cobalt chloride CoCl2 (100 μM) to simulate the in vitro hypoxic environment;
[0042] (2) Drug treatment: add HIF-1a inhibitor CAY10585 (0.7 μM) to the supernatant of the well to stimulate gastric cancer cells
[0043] (3) Use colorimetric method to detect the change of glucose uptake rate and lactic acid production rate of gastric cancer cells, reflecting the change of glucose metabolism reprogramming of cells
[0044] 2. In vivo treatment experiment (gastric cancer orthotopic tumor model treatment experiment):
[0045] Material source: HIF-1a inhibitor CAY10585 was purchased from Selleck Company (product number S8441), apatinib was purchased from Selleck Company (product number S5248)
[0046] Experimental steps:
[0047] (1) Gastric cancer orthotopic implantation tumor model construction:
[0048] (a) Prepare 4-6 week old athymic nude mice, male, body weight 18-20 g, fed in SPF level environment;
[0049] (b) Construction of subcutaneous tumor model in nude mice: well-conditioned gastric cancer cells were prepared into a single cell suspension (5 x 10 7 After 75% alcohol disinfection, the cell suspension was injected into the right groin of the nude mice (subcutaneous injection, 200 μl per mouse), and the tumor growth was observed regularly after inoculation of gastric cancer cells;
[0050] (c) When the subcutaneous tumor grows to 1.0-1.5 cm, the nude mice were euthanized, the tumor mass was peeled off and cut into several pieces (about 1 mm 3 ) in size with a sterile blade, and placed in Hanks buffer for standby;
[0051] (d) Orthotopic implantation: after anesthesia with ether inhalation and alcohol disinfection of the surgical area skin, the abdominal wall of the nude mice was incised along the left midline and the peritoneum and gastric wall were carefully exposed; the tumor mass was sutured to the gastric wall using absorbable suture, and the abdomen was sutured;
[0052] (2) Group design and drug treatment:
[0053] The successfully constructed nude mice were randomly divided into control group (Control), apatinib treatment group, apatinib+CAY10585 treatment group, 6 mice per group; On the 10th day after orthotopic implantation, the nude mice were treated with apatinib (120 mg / kg daily, gavage; apatinib treatment group and apatinib+CAY10585 treatment group), and the control group (Control) of nude mice was treated with an equal volume of DMSO solution; On the 24th day after operation, the nude mice were treated with CAY10585 (10 mg / kg daily, gavage; apatinib+CAY10585 treatment group), and the control group (Control) and apatinib treatment group of nude mice were treated with an equal volume of DMSO solution;
[0054] (3) Observation of the growth and metastasis of in situ implanted tumors: The nude mice were killed by dislocation at 8 weeks after surgery, and the thoracic and abdominal cavities were fully explored. The in situ implanted tumors in the abdominal cavity were removed, the size of the tumors was measured, and the tumors were fixed with formalin and then stained with hematoxylin-eosin and immunohistochemically. At the same time, the metastasis of the tumors in the thoracic and abdominal cavities of the nude mice (such as metastasis to adjacent organs) was observed.
[0055] 2. Experimental Results
[0056] 1. The hypoxic microenvironment induced by anti-angiogenic therapy promotes HIF- accumulation ( Figure 2 ).
[0057] The analysis of the in vitro hypoxia model of cobalt chloride (CoCl2) showed that the higher the degree of cell hypoxia (CoCl2 concentration), the higher the intracellular expression level of HIF-1a ( Figure 2 Middle A). Immunofluorescence detection of human gastric cancer tissues revealed that HIF-1a was significantly enriched in gastric cancer tissues in hypoxic areas ( Figure 2 Apatinib was further used to construct an anti-angiogenic therapy resistance model, inducing a hypoxic microenvironment in subcutaneous tumors in mice ( Figure 2 Middle C), immunofluorescence results showed that the neovascularization in the subcutaneous tumors of mice treated with apatinib was significantly reduced, and there was significant hypoxia in the subcutaneous tumors, and HIF-1a expression was significantly increased ( Figure 2 Middle D).
[0058] 2. Hypoxia-inducible factor HIF-1a improves cell hypoxia tolerance by enhancing cellular glucose metabolism reprogramming ( Figure 3 )
[0059] Glucose metabolic reprogramming is a key intrinsic mechanism of tumor progression and a characteristic metabolic alteration of tumors. Normal cells maintain a balanced glucose metabolism, primarily synthesizing adenosine triphosphate (ATP) through mitochondrial oxidative phosphorylation. However, tumor cells, on the other hand, are in a state of hyperglycemic glucose metabolism. This process, known as glucose metabolic reprogramming, shifts cellular energy supply through the "Warburg effect," shifting glucose metabolism from mitochondrial oxidative phosphorylation to glycolysis.
[0060] Using the CoCl2 in vitro hypoxia model, the intracellular level of HIF-1a was significantly upregulated after cell hypoxia ( Figure 3 Middle A), the glucose uptake rate and lactate production level of gastric cancer cells were significantly upregulated; and the higher the degree of hypoxia, the higher the glycolysis level of gastric cancer cells ( Figure 3 At the same time, the mitochondrial membrane potential of gastric cancer cells decreased significantly under hypoxia, and mitochondrial function was inhibited ( Figure 3Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 3 Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells.
[0061] Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 4 )
[0062] Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 4 Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 4 Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells.
[0063] Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 5 )
[0064] Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 1 Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 1 Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. Figure 5 Figure 6. CAY10585 effectively inhibits glucose metabolic reprogramming and attenuates hypoxia tolerance of gastric cancer cells. (A) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells. (B) CAY10585 effectively attenuates hypoxia tolerance of gastric cancer cells. (C) CAY10585 effectively inhibits mitochondrial function of gastric cancer cells. (D) and (E) CAY10585 effectively inhibits glucose metabolic reprogramming of gastric cancer cells.Figure 5 In summary, the animal experiments confirmed that the combination treatment of apatinib and CAY10585 can effectively inhibit the growth and metastasis of gastric cancer orthotopic xenografts and enhance the anti-tumor effect.
[0065] Although the above embodiments have made a detailed description of the present application, it is only a part of the embodiments of the present application, not all the embodiments, and other embodiments can be obtained according to the present embodiments without creativity, which are within the protection scope of the present application.
Claims
1. A pharmaceutical composition for treating gastric cancer, characterized in that: The active ingredients include apatinib and CAY10585 at a working concentration ratio of 120 mg / kg: 10 mg / kg.
2. Use of the pharmaceutical composition of claim 1 in the preparation of a medicament for treating gastric cancer.
3. A medicament for treating gastric cancer, characterized by comprising the compound of claim 1 or 2. The active ingredients include the pharmaceutical composition of claim 1 and pharmaceutically acceptable adjuvants.
4. The medicament according to claim 3, characterized in that, The active ingredients have a mass percentage of 0.1-99%.
5. The medicament according to claim 3, characterized in that, The dosage form of the medicament includes oral, intravenous or intraperitoneal injection.