Application of compound in preparation of medicine for inhibiting G3BPs

Abstract

The invention provides application of a compound in preparation of a medicine for inhibiting G3BPs, and belongs to the technical field of biological medicine. The medicine is used for treating tumors, neurodegenerative diseases or virus infection; the tumors comprise any one or a combination of lung cancer, breast cancer, colorectal cancer, prostate cancer, stomach cancer, liver cancer, pancreatic cancer, cervical cancer, skin cancer, osteosarcoma, liposarcoma, smooth muscle sarcoma, leukemia, lymphoma, multiple myeloma, glioma, meningioma, testicular cancer, endometrial cancer, cervical cancer, ovarian cancer, melanoma, thyroid cancer, kidney cancer or bladder cancer. When the compound shown in the formula (I) is combined with different anti-tumor drugs in various tumor cells, the sensitivity of the cells to the anti-tumor drugs can be improved, so that the drug resistance of the tumor cells is reduced.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the use of a compound in the preparation of a drug for inhibiting G3BPs. Background Technology

[0002] G3BPs (Ras-GTPase-activating protein-binding proteins) are RNA-binding proteins that play a crucial role in RNA processing, degradation, and the formation of stress granules (SGs). There are two homologous proteins of G3BPs: G3BP1 and G3BP2. Due to the high degree of redundancy in their structure and function, the function of "G3BP1" in the following text will encompass that of G3BP2.

[0003] G3BP1 is overexpressed in various human tumors, promoting cancer cells to enter the DNA synthesis phase (S phase) and accelerating cell growth. It plays a crucial role in cancer cell proliferation, differentiation, and apoptosis, and is closely related to cancer cell drug resistance. Furthermore, G3BP1 participates in multiple carcinogenesis-related signaling pathways, including NF-κB, ERK, p53, and Ras. G3BP1 is involved in the infection process of various viruses; some viruses target G3BP1 to inhibit cellular responses, and interactions between viral proteins or RNA and host G3BP1 proteins can disrupt the cell's innate immune response. In addition, G3BP1 dysfunction in neurodegenerative diseases leads to abnormal solidification of SGs and co-aggregation with disease proteins, thereby driving neurotoxicity. In conclusion, G3BP1 is a promising new target for the treatment of cancer, viral infections, and neurodegenerative diseases, and designing a substance that can inhibit G3BP1 has broad application prospects.

[0004] The invention patent application CN120324399A discloses the application of compound UA-0007373 (referred to as compound (I) in this application) in the preparation of tumor therapeutic drugs. This small molecule inhibitor can effectively inhibit the formation of cellular stress granules induced by G3BP1 or anticancer drugs, thereby reducing the drug resistance of tumor cells. However, this document only verifies the therapeutic effect of compound UA-0007373 in human breast cancer cells when used in combination with the chemotherapy drug sorafenib. Regarding the therapeutic effects of this compound on other tumors, and its effects when used in combination with other anticancer drugs, this document does not provide any experimental results. Summary of the Invention

[0005] In view of this, in order to at least partially solve the aforementioned technical problems, the present invention provides the use of a compound in the preparation of a drug for inhibiting G3BPs.

[0006] According to one aspect of the present invention, there is provided the use of a compound of formula (I) in the preparation of a medicament for inhibiting G3BPs, the medicament being used to treat tumors, neurodegenerative diseases, or viral infections; wherein the tumors include any one or a combination of lung cancer, breast cancer, colorectal cancer, prostate cancer, gastric cancer, liver cancer, pancreatic cancer, cervical cancer, skin cancer, osteosarcoma, liposarcoma, leiomyosarcoma, leukemia, lymphoma, multiple myeloma, glioma, meningioma, testicular cancer, endometrial cancer, cervical cancer, ovarian cancer, melanoma, thyroid cancer, kidney cancer, or bladder cancer; wherein the neurodegenerative diseases include acute neurodegenerative diseases and chronic neurodegenerative diseases, wherein chronic neurodegenerative diseases include any one of Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, or frontotemporal dementia, and acute neurodegenerative diseases include any one of cerebral ischemia, brain injury, or epilepsy;

[0007] (I).

[0008] The compound of formula (I) of this invention, when used to treat tumors, can increase the sensitivity of cells to anti-tumor drugs, thereby reducing the drug resistance of tumor cells. Therefore,

[0009] According to one embodiment of the present invention, the above-mentioned drug is used to reduce the drug resistance of tumor cells to anti-tumor drugs.

[0010] According to one embodiment of the present invention, the above-mentioned antitumor drug includes any one or a combination of 5-fluorouracil, paclitaxel, doxorubicin, platinum-based drugs, oxaliplatin, bortezomib, docetaxel or cabazitaxel.

[0011] According to another aspect of the present invention, a pharmaceutical combination is provided, the pharmaceutical combination comprising a compound of formula (I) above and an antitumor drug, said antitumor drug comprising any one or a combination of 5-fluorouracil, paclitaxel, doxorubicin, platinum-based drugs, oxaliplatin, bortezomib, docetaxel or cabazitaxel.

[0012] The compound of formula (I) of this invention can bind to the NTF2L domain of G3BP1, thereby affecting the normal physiological function of G3BP1. When used in combination with antitumor drugs, the compound of formula (I) can enhance the sensitivity of cells to antitumor drugs, thereby reducing the drug resistance of tumor cells. Attached Figure Description

[0013] Figure 1 The half-maximal inhibitory concentration (IC50) of compound (Ⅰ) in combination with sorafenib in hepatocellular carcinoma cells (Huh7) is given. 50 ) Test result image;

[0014] Figure 2The IC50 of compound (I) in combination with paclitaxel in breast cancer cells (MDA-MB-231) is given. 50 The test result image shows that 231 represents MDA-MB-231.

[0015] Figure 3 The IC50 of compound (I) in combination with doxorubicin in MDA-MB-231 50 The test result image shows that 231 represents MDA-MB-231.

[0016] Figure 4 The IC50 of compound (I) in combination with cisplatin in lung cancer cells (H1299) 50 Test result image;

[0017] Figure 5 The IC50 of compound (I) in combination with paclitaxel in gastric cancer cells (HGC27) is given. 50 Test result image;

[0018] Figure 6 The IC50 of compound (I) in combination with paclitaxel in osteosarcoma cells (U2OS) 50 Detection results image. Detailed Implementation

[0019] In realizing the concept of this invention, it was discovered that multiple proteins, including Caprin-1, USP10, SARS-CoV-2 N protein, and SFVnsP3, can collectively recognize and bind to the G3BP1 NTF2L domain. Occupying the G3BP1 NTF2L domain can affect the normal physiological function of G3BP1 and may affect its function in diseases such as cancer, viral infections, and neurodegenerative diseases. Therefore, compounds targeting the G3BP1 NTF2L domain were screened for binding. The screened compounds, by specifically occupying the NTF2L domain of G3BP1, directly interfered with the core molecular function of G3BP1 as an RNA-binding protein. Its mechanism of action mainly involves: competitively inhibiting the binding of G3BP1 to downstream target mRNAs (such as oncogene transcripts, viral genomes, or abnormal transcripts), leading to decreased stability and translational dysregulation of these mRNAs; and blocking the interaction of G3BP1 with key signaling proteins (such as Ras GTPase activator proteins or innate immune aptamers), thereby dismantling its scaffold function in disease-related pathways. These effects collectively achieve broad inhibition of abnormal G3BP1 function in various pathological environments such as cancer, viral infection, and neurodegenerative diseases, providing a new targeted strategy for intervening in these diseases.

[0020] Because G3BP1 is involved in the infection process of various viruses and can drive neurotoxicity in neurodegenerative diseases, it is a potential new target for viral infections and neurodegenerative diseases. The aforementioned neurodegenerative diseases cover both acute and chronic categories. Chronic types, with a longer course and slower progression, mainly include Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), and frontotemporal dementia (FTD). Acute types, with a rapid onset and rapid progression, include cerebral ischemia (CI), brain injury (BI), and epilepsy.

[0021] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. However, it should be understood that these descriptions are merely exemplary and not intended to limit the scope of the invention.

[0022] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0023] Example (I) shows how compounds of formula (I) can be used in combination with different antitumor drugs to enhance the sensitivity of various tumor cells to the drugs.

[0024] (1) Cell culture: Huh7, MDA-MB-231, H1299, HGC27 and U2OS were cultured in 9 cm culture dishes using DMEM medium containing 10% (v / v) fetal bovine serum and cultured in a constant temperature incubator at 37℃ and 5% CO2.

[0025] (2) Cell washing: When the cell confluence reaches 80%, the culture medium is discarded and phosphate-buffered saline (PBS) is added to gently wash the cells.

[0026] (3) Pancreatic enzyme digestion: After discarding PBS, add 2 mL of pancreatic enzyme and digest at 37°C for 3 min.

[0027] (4) Cell collection: Transfer the cell suspension to an EP tube, centrifuge at 1200×g for 5 min, and discard the supernatant.

[0028] (5) Cell seeding: Resuspend the cells in 1 mL of DMEM and dilute to a density of 4000 cells / mL. Seed the cells in 96-well plates and culture for 24 h to allow the cells to adhere completely. At this time, the cell confluence in each well reaches 50%-70%.

[0029] (6) Treatment with compound (I): An experimental group (combined drug group) and a control group (anti-tumor drug monotherapy group) were set up. Compound (I) solution was added to the wells treated with compound (I) (experimental group) to make the final concentration of compound (I) in the wells 25 μM or 50 μM; no compound (I) treatment was performed in the control wells (control group).

[0030] (7) After the cells treated with the compound of formula (Ⅰ) are incubated in an incubator for 1 hour, anti-tumor drugs are added.

[0031] (8) Treatment with antitumor drugs:

[0032] a. Sorafenib administration in Huh7: 10 μL of serially diluted sorafenib was administered to the corresponding positions in a 96-well plate containing Huh7 (both experimental and control groups were administered the drug), so that the final drug concentrations were 200, 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, and 0.78125 μM, with 6 replicates per group;

[0033] b. Administration of paclitaxel or doxorubicin in MDA-MB-231: Take 10 μL of serially diluted paclitaxel or doxorubicin into the corresponding positions of a 96-well plate coated with MDA-MB-231 (both experimental and control groups need to be administered), so that the final drug concentrations are 2000, 1000, 400, 200, 100, 50, 25, 12.5, 6.25, and 3.125 nM, with 6 replicates per group;

[0034] c. Cisplatin administration in H1299: 10 μL of serially diluted cisplatin was administered to the corresponding positions in a 96-well plate containing H1299 (both the experimental and control groups were administered the drug), so that the final drug concentrations were 200, 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, and 0.78125 μM, with 6 replicates per group;

[0035] d. Paclitaxel administration in HGC27: 10 μL of serially diluted paclitaxel was administered to the corresponding positions in a 96-well plate coated with H1299 (both experimental and control groups were administered the drug), so that the final drug concentrations were 2000, 1000, 400, 200, 100, 50, 25, 12.5, 6.25, and 3.125 nM, with 6 replicates per group;

[0036] f. Paclitaxel administration in U2OS: 10 μL of serially diluted paclitaxel was administered to the corresponding positions in a 96-well plate coated with U2OS (both experimental and control groups were administered the drug), so that the final drug concentrations were 2000, 1000, 400, 200, 100, 50, 25, 12.5, 6.25, and 3.125 nM, with 6 replicates per group.

[0037] (9) Drug incubation: The cells of the control group and the experimental group were placed in a 37℃, 5% CO2 incubator and cultured for 72h.

[0038] (10) CCK8 detection: Discard the original culture medium, add 100 μL of a mixture of CCK8 reagent (Beyotime C0038) and DMEM culture medium (1:10) to each well, and incubate in a 37℃ incubator in the dark for 1 h.

[0039] (11) Data analysis: The absorbance of cells at 450 nm was detected using an ELISA reader, and the IC50 of the antitumor drug combined with the compound of formula (I) was calculated using data analysis software. 50 and the effect of antitumor drugs on the IC50 of cells 50 .

[0040] a. The inhibitory effect of compound (I) in combination with sorafenib on Huh7

[0041] The inhibitory effect of compound (I) in combination with sorafenib on Huh7 is as follows: Figure 1 As shown. Sorafenib alone for Huh7 IC 50 The concentration was 24.78 μM, while the IC50 of sorafenib was significantly higher after combining it with 25 μM of compound (I). 50 It decreased to 12.87 μM (a 48% reduction).

[0042] The above results indicate that compound (I) targets the binding of G3BP1 and significantly enhances the sensitivity of Huh7 to the anti-tumor drug sorafenib. Specifically, compound (I) increases the killing efficacy of sorafenib against Huh7 by 1.9 times (IC50). 50 (From 24.78 to 12.87 μM).

[0043] b. Inhibitory effect of compound (I) in combination with paclitaxel or doxorubicin on MDA-MB-231

[0044] The inhibitory effect of compound (I) in combination with paclitaxel on MDA-MB-231 cells is as follows: Figure 2 As shown. The IC50 of paclitaxel alone on MDA-MB-231 cells. 50 The IC50 of paclitaxel was 29.25 nM, while the IC50 of paclitaxel was higher after combining it with 50 μM of compound (Ⅰ). 50 It decreased to 12.63 nM (a 57% reduction).

[0045] The inhibitory effect of compound (I) in combination with doxorubicin on MDA-MB-231 cells is as follows: Figure 3 As shown. The IC50 of doxorubicin alone on MDA-MB-231 cells. 50The IC50 of doxorubicin was 182.3 nM, while the IC50 of doxorubicin after combining with 50 μM of compound (I) was... 50 It decreased to 92.75 nM (a 49% reduction).

[0046] The above results indicate that compound (I) can target G3BP1 and significantly enhance the sensitivity of MDA-MB-231 to anti-tumor drugs. Specifically, compound (I) increases the killing efficacy of paclitaxel against U2OS by 2.3 times (IC50). 50 From 29.25 → 12.63 nM); Compound (I) increased the killing efficacy of doxorubicin against U2OS by 2 times (IC50). 50 From 182.3 → 92.75 nM).

[0047] c. Inhibitory effect of compound (I) in combination with cisplatin on lung cancer cells H1299

[0048] The inhibitory effect of compound (I) in combination with cisplatin on H1299 is as follows: Figure 4 As shown. Cisplatin alone is used for H1299 IC. 50 The concentration of cisplatin was 15.6 μM, while the concentration of cisplatin in H1299 cells was significantly higher when combined with 50 μM of the compound of formula (I). 50 It decreased to 4.635 μM (a reduction of 70.2%).

[0049] The above results indicate that compound (I) targets the binding of G3BP1 and significantly enhances the sensitivity of H1299 cells to the anti-tumor drug cisplatin. Specifically, compound (I) increases the killing efficacy of cisplatin against H1299 cells by 3.4 times (IC50). 50 From 15.6 to 4.635 μM.

[0050] d. Inhibitory effect of compound (Ⅰ) in combination with paclitaxel on HGC27

[0051] The inhibitory effect of compound (I) in combination with paclitaxel on HGC27 is as follows: Figure 5 As shown. The effect of paclitaxel alone on the IC50 of HGC27 cells. 50 The IC50 of paclitaxel on HGC27 cells was 59.62 nM, while the IC50 of paclitaxel on HGC27 cells was significantly lower after combining with 50 μM of compound (Ⅰ). 50 It decreased to 3.21 nM (a reduction of 94.6%), which is a very significant effect.

[0052] The above results indicate that compound (I) can target G3BP1 and significantly enhance the sensitivity of HGC27 to the anti-tumor drug paclitaxel. Specifically, compound (I) increases the killing efficacy of paclitaxel against HGC27 by 18.6 times (IC50). 50 From 59.62 to 3.21 nM).

[0053] e. The inhibitory effect of compound (I) in combination with paclitaxel on U2OS

[0054] The inhibitory effect of compound (I) in combination with paclitaxel on U2OS is as follows: Figure 6 As shown. The IC of U2OS using paclitaxel alone. 50 The IC50 of paclitaxel against U2OS was 78.19 nM, while the IC50 of paclitaxel against U2OS was 78.19 nM after combining with 50 μM of compound (Ⅰ). 50 It decreased to 21.85 nM (a 72% reduction).

[0055] The above results indicate that compound (I) can target G3BP1 and significantly enhance the sensitivity of U2OS to the anti-tumor drug paclitaxel. Specifically, compound (I) increases the killing efficacy of paclitaxel against U2OS by 3.6 times (IC50). 50 From 78.19 → 21.85 nM).

[0056] In summary, compounds of formula (I) targeting G3BP1, when combined with different antitumor drugs (sorafenib, paclitaxel, doxorubicin, and cisplatin), can effectively enhance the drug sensitivity of various tumor cells (Huh7, MDA-MB-231, H1299, HGC27, and U2OS), demonstrating the potential to overcome drug resistance in various tumors. Therefore, compounds of formula (I) can serve as novel chemosensitizers, overcoming drug resistance in cancers such as liver cancer, breast cancer, lung cancer, gastric cancer, and osteosarcoma through G3BP1 targeting. Similarly, compounds of formula (I) can also serve as promising small molecule drugs for treating other G3BP1-related diseases, such as neurodegenerative diseases and viral infections.

[0057] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are 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 within the protection scope of the present invention.

Claims

1. The use of the compound of formula (I) in the preparation of a drug for inhibiting G3BPs, characterized in that, The drug is used to treat tumors, neurodegenerative diseases, or viral infections; The tumors include any one or a combination of lung cancer, breast cancer, colorectal cancer, prostate cancer, stomach cancer, liver cancer, pancreatic cancer, cervical cancer, skin cancer, osteosarcoma, liposarcoma, leiomyosarcoma, leukemia, lymphoma, multiple myeloma, glioma, meningioma, testicular cancer, endometrial cancer, cervical cancer, ovarian cancer, melanoma, thyroid cancer, kidney cancer, or bladder cancer. The neurodegenerative diseases mentioned include acute neurodegenerative diseases and chronic neurodegenerative diseases; The chronic neurodegenerative diseases include any one of Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, or frontotemporal dementia; the acute neurodegenerative diseases include any one of cerebral ischemia, brain injury, or epilepsy. （Ⅰ）。 2. The application as described in claim 1, wherein the drug is used to reduce the resistance of tumor cells to anti-tumor drugs.

3. The application as described in claim 2, wherein the antitumor drug comprises any one or a combination of 5-fluorouracil, paclitaxel, doxorubicin, platinum-based drugs, oxaliplatin, bortezomib, docetaxel, or cabazitaxel.

4. A drug combination, characterized in that, The drug combination comprises a compound of formula (I) as described in claim 1 and an antitumor drug, wherein the antitumor drug comprises any one or a combination of 5-fluorouracil, paclitaxel, doxorubicin, platinum-based drugs, oxaliplatin, bortezomib, docetaxel or cabazitaxel.

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