Combined application of G6PD inhibitor and Fatostatin in treatment of brain glioma
Through the pharmaceutical composition of G6PD inhibitor and Fatostatin, the problem of short effective period and obvious side effects in the treatment of brain glioma is solved, and the effect of significantly inhibiting brain glioma cell growth and promoting cell death is achieved. Research on the treatment and metabolic regulation mechanism of glioma provides new ideas and tools.
Patent Information
- Application Number
- CN202411680524.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-20
- Publication Date
- 2025-02-07
AI Technical Summary
The prior art has problems in the treatment of brain gliomas with short effective periods, obvious side effects and inability to improve overall survival, especially the lack of effective metabolic regulatory mechanisms and new molecular targets.
The pharmaceutical composition of G6PD inhibitor and Fatostatin is used to inhibit the growth of brain glioma cells and promote cell death through synergistic effects.
This pharmaceutical composition can significantly enhance the sensitivity of cancer cells to lipid synthesis inhibitors, provide new effective molecular targets, improve the therapeutic effect of brain gliomas, and provide new ideas for the study of metabolic regulatory mechanisms.
Smart Images
Figure CN119386185A_ABST
Abstract
Description
[0001] This application is a divisional application, and the parent case information is as follows: The name of the invention is combined use of G6PD inhibitors and inhibitors of new molecular points; the original application date is 2023.10.20, and the original application number is 2023113640359. Technical Field
[0002] The invention belongs to the field of medicine and biotechnology, and particularly relates to the combined application of a G6PD inhibitor and Fatostatin to treat brain glioma. Background Art
[0003] Glioblastoma Multiform (GBM) is a WHO grade IV glioma with a median survival of only 12-15 months after diagnosis. It is the malignant tumor with the highest mortality rate in the world. In addition, the prognosis of GBM patients has not improved significantly in the past few decades, mainly due to the lack of effective treatment strategies.
[0004] The standard treatment for GBM is to first perform maximal surgical resection, followed by simultaneous radiotherapy and temozolomide (TMZ) for 6 weeks, followed by continued TMZ treatment every 4 weeks for 6 cycles. However, this treatment is only effective for a few months, and almost all GBM tumors inevitably recur after treatment. In addition to TMZ, only bevacizumab is currently approved by the FDA for the clinical treatment of recurrent GBM. However, bevacizumab can only effectively relieve symptoms and improve quality of life in the short term, and bevacizumab has no combined effect with TMZ and radiotherapy; in addition, it reduces the absorption of TMZ, cannot improve overall survival, and has significant side effects such as hypertension, venous thrombosis, and infection.
[0005] Recent studies have shown that metabolic reprogramming is one of the core characteristics of tumors. Exploring the regulatory mechanism of GBM metabolism, analyzing its biological characteristics, and finding effective molecular targets to guide clinical treatment can provide new ideas and efficient targets for cutting off GBM energy supply. Summary of the invention
[0006] In order to make up for the deficiencies of the prior art, the object of the present invention is to provide a pharmaceutical composition of a G6PD inhibitor and Fatostatin and its application in the preparation of a product for treating brain glioma.
[0007] In order to achieve the above objectives, the present invention adopts the following technical scheme: In a first aspect, the present invention provides a pharmaceutical composition for treating brain glioma, the pharmaceutical composition comprising: (i) a therapeutically effective amount of a G6PD inhibitor; (ii) a therapeutically effective amount of a SREBP1 inhibitor.
[0008] Furthermore, the G6PD inhibitor is G6PDi-1 or a pharmaceutically acceptable salt thereof.
[0009] Specifically, the G6PDi-1 is a reversible non-competitive 6-phosphate glucose dehydrogenase inhibitor with an IC50 of 0.07 μM and a structural formula of:
[0010] Furthermore, the SREBP1 inhibitor is Fatostatin or a pharmaceutically acceptable salt thereof.
[0011] In the present invention, the pharmaceutically acceptable salt refers to a salt of an active compound (such as G6PDi-1) and is prepared by reacting the active compound with a suitable organic or inorganic acid or acid derivative. Pharmaceutically acceptable salts include, but are not limited to, hydrochloride, sulfate, phosphate, citrate, hydrobromide, acetate, benzoate, benzenesulfonate, tartrate, carbonate, citrate, gluconate, lactate, malate, methanesulfonate, stearate, valerate, nitrate, sodium salt, calcium salt, potassium salt, zinc salt and meglumine salt.
[0012] Furthermore, the dosage ratio of the G6PD inhibitor to the SREBP1 inhibitor is 1:0.1-2.
[0013] Furthermore, the dosage ratio of the G6PD inhibitor to the SREBP1 inhibitor is selected from 3:1, 2:1, 1:1, 1:2, and 1:3.
[0014] Furthermore, the G6PD inhibitor and the SREBP1 inhibitor are present in the same or different compositions or kits.
[0015] Furthermore, the use of the G6PD inhibitor and the SREBP1 inhibitor includes concurrent administration or sequential administration. Specifically, the administration is to provide therapeutically effective levels of the G6PD inhibitor and the SREBP1 inhibitor in vivo.
[0016] In the present invention, the term "effective amount" or "therapeutically effective level" refers to a dose that can achieve treatment of brain glioma in a subject. It depends on the patient, the severity of the disease, and the route and medium used for administration. The amount of active compound in such a therapeutic composition is a suitable dose.
[0017] Furthermore, the pharmaceutical composition also includes a pharmaceutically acceptable carrier or excipient. Specifically, the pharmaceutically acceptable carrier or excipient is a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or any type of formulation aid.
[0018] In the present invention, the term "pharmaceutically acceptable carrier" refers to any pharmaceutical carrier that does not itself induce the production of antibodies that are harmful to the individual receiving the composition and can be administered without excessive toxicity. Suitable carriers can be large, slowly metabolized macromolecules, such as proteins, polysaccharides, polylactic acid, polyglycolic acid, polymeric amino acids and amino acid copolymers. Such carriers are well known to those of ordinary skill in the art. The pharmaceutically acceptable carrier in the pharmaceutical composition can include fluids, such as water, saline, glycerol and ethanol. Auxiliary substances, such as wetting agents or emulsifiers, pH buffer substances, etc., can also be present in such vehicles.
[0019] Furthermore, the brain glioma includes grade I, grade II, grade III, and grade IV brain glioma.
[0020] Furthermore, the glioma is a grade IV glioma.
[0021] Furthermore, the brain glioma is glioblastoma multiforme.
[0022] Furthermore, the dosage forms of the G6PD inhibitor and the SREBP1 inhibitor may be the same or different.
[0023] Furthermore, the dosage form includes drops, tablets, capsules, granules, films, gels, powders, emulsions, pills, suppositories, aerosols, sprays, powder mists, patches, solutions, ointments or creams.
[0024] In the present invention, the term "pharmaceutical composition" refers to a composition comprising at least one biologically active compound. The pharmaceutical composition of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or by implanted drug storage devices. In some embodiments, oral administration is preferred. The pharmaceutical composition of the present invention may contain any commonly used non-toxic pharmaceutically acceptable carrier, adjuvant or vehicle. In some cases, a pharmaceutical acid, base or buffer may be used to adjust the pH of the formulation to improve the stability of the formulated compound or its dosage form. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques. As long as the target tissue can be reached, the pharmaceutical composition of the present invention may be administered to the receptor by any route.
[0025] The second aspect of the present invention provides use of the pharmaceutical composition described in the first aspect of the present invention in the preparation of a product for treating brain glioma.
[0026] Furthermore, the product includes a test kit, a chip, a test strip, or a device, an equipment, or a computer-readable storage medium.
[0027] Furthermore, the product may also contain other drugs for treating cancer.
[0028] The other drugs for treating cancer are Acivicin, Aclarubicin, Acodazole Hydrochloride, AcrQnine, Adozelesin, Aldesleukin, Asparaginase, Asperlin, Azacitidine, Azetepa, Azotomycin, Batimastat, Benzodepa, Bicalutamide, Bisantrene Hydrochloride, Bisnafide Dimesylate, Bizelesin, Bleomycin Sulfate, Brequinar Sodium Sodium), Bropirimine, Busulfan, Cactinomycin, Calusterone, Caracemide, Carbetimer, Carboplatin, Carmustine, Carubicin Hydrochloride, Carzelesin, Cedefingol, Chlorambucil, Cirolemycin, Cisplatin, Cladribine, etc.
[0029] Furthermore, the product may also contain other drugs for treating brain glioma.
[0030] In the present invention, the term "treatment" refers to the process of intervening or changing a specific health state, including eliminating the cause, symptomatic treatment or supportive treatment, and does not necessarily mean curing or completely eliminating the disease, condition or symptoms of the disease or condition. Treatment may refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in the severity of the symptoms / effects of an established disease.
[0031] Furthermore, the brain glioma includes grade I, grade II, grade III, and grade IV brain glioma.
[0032] Furthermore, the glioma is a grade IV glioma.
[0033] Furthermore, the brain glioma is glioblastoma multiforme.
[0034] The third aspect of the present invention provides the use of a reagent for detecting the expression level of G6PD, the use comprising any one of the following: 1) use in the preparation of a product for diagnosing brain glioma; 2) use in the preparation of a product for grading brain glioma; 3) use in the preparation of a product for predicting the prognosis of brain glioma.
[0035] Specifically, the G6PD expression level in GBM is higher than that in normal brain tissue.
[0036] Furthermore, the detection is performed on samples from patients or subjects, and the samples include tissues, cells, blood, urine, saliva, mucosal samples, feces, intestinal lavage, joint fluid, cerebrospinal fluid, bile samples, respiratory secretions, and bronchoalveolar lavage fluid samples.
[0037] In the present invention, the term subject can be a vertebrate, more specifically a mammal (e.g., a human, a horse, a pig, a rabbit, a dog, a sheep, a goat, a non-human primate, a cow, a cat, a guinea pig or a rodent), a fish, a bird or a reptile or an amphibian. The term does not indicate a specific age or gender. Therefore, the subject covers adult subjects and newborn subjects, regardless of gender. In the context of the present invention, patient or subject are used interchangeably to refer to a subject suffering from / suspected of having a disease. The disease targeted by the present invention is a patient with a glioma, preferably a grade IV glioma, i.e., glioblastoma multiforme (GBM, also known as glioblastoma).
[0038] Furthermore, the sample is tissue or cell.
[0039] Furthermore, the brain glioma includes grade I, grade II, grade III, and grade IV, and the expression level of G6PD increases with the grade.
[0040] Furthermore, the glioma is a grade IV glioma.
[0041] Furthermore, the brain glioma is glioblastoma multiforme.
[0042] Furthermore, the reagent for detecting the expression level of G6PD includes a reagent for detecting the expression level of G6PD protein and / or the expression level of G6PD mRNA.
[0043] Furthermore, the reagents for detecting the expression amount of G6PD protein include reagents used in the following methods: hematoxylin-eosin staining, safranin O-fast green staining, protein blotting, enzyme-linked immunosorbent assay, radioimmunoassay, sandwich assay, immunohistochemical staining method, mass spectrometry, immunoprecipitation analysis, complement fixation analysis, flow cytometry fluorescence sorting technology and protein chip method.
[0044] Furthermore, the reagents for detecting the expression of G6PD mRNA include reagents used in the following methods: PCR-based detection method, Southern hybridization method, Northern hybridization method, dot hybridization method, fluorescence in situ hybridization method, DNA microarray method, ASO method, and high-throughput sequencing platform method.
[0045] Advantages and beneficial effects of the present invention: The pharmaceutical composition provided by the present invention can synergistically enhance efficacy, can be used to prepare products for treating gliomas, and provides new effective molecular targets for gliomas, providing research tools and new ideas for studying the metabolic regulation mechanism of gliomas. BRIEF DESCRIPTION OF THE DRAWINGS
[0046] Figure 1 This is the comparison result of G6PD expression in GBM and normal brain tissue.
[0047] Figure 2 This is the comparison result of the expression level of G6PD in gliomas of different grades.
[0048] Figure 3 This is the survival curve of G6PD in GBM.
[0049] Figure 4 This is the result of using tissue chips to analyze the expression of G6PD in gliomas of different grades.
[0050] Figure 5 It is a survival curve drawn based on the results of tissue chip detection.
[0051] Figure 6 This is the result of the correlation analysis between G6PD and other genes.
[0052] Figure 7 It is the expression result of SREBP-1 and G6PD in tumor tissues of GBM patients.
[0053] Figure 8 This is a graph showing the cell number statistics of U87 cells treated with the SREBP1 inhibitor Fatostatin in combination with G6PDi-1.
[0054] Fig. 9This is a microscopic image of U87 cells treated with the SREBP1 inhibitor Fatostatin in combination with G6PDi-1.
[0055] Fig.10 This is a graph showing the cell number statistics of U251 cells treated with the DGAT1 inhibitor A922500 in combination with G6PDi-1.
[0056] Fig.11 These are microscope images of U251 cells treated with the DGAT1 inhibitor A922500 in combination with G6PDi-1.
[0057] Fig.12 This is a graph showing the colony formation results of DGAT1 inhibitor A922500 and G6PDi-1.
[0058] Fig.13 This is a graph showing the cell number statistics of U251 cells treated with the GPX4 inhibitor RSL-3 in combination with G6PDi-1.
[0059] Fig.14 These are microscope images of U251 cells treated with the GPX4 inhibitor RSL-3 and G6PDi-1. DETAILED DESCRIPTION
[0060] The present invention is further described below in conjunction with the embodiments. The following description is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any technician familiar with the profession may use the above disclosed technical content to change it into an equivalent embodiment with equivalent changes. Any simple modification or equivalent change made to the following embodiments based on the technical essence of the present invention without departing from the content of the present invention falls within the protection scope of the present invention.
[0061] Example 1. Role of G6PD in the diagnosis, grading, and prognosis of GBM By analyzing RNAseq data of 32 solid tumors from The Cancer Genome Atlas (TCGA), it was found that the expression of G6PD in GBM was significantly higher than that in normal brain tissue ( Figure 1 ); the expression in GBM was also significantly higher than that in low-grade glioma (LGG) ( Figure 2 High expression of G6PD was significantly associated with poor survival in the GBM cohort ( Figure 3 ).
[0062] Accordingly, we further detected the G6PD protein level in tumor tissues of patients with grade I-IV gliomas using tissue microarray (TMA) (n=145). IHC staining results showed that the G6PD protein content in grade IV GBM tissues was the highest compared with grade III gliomas, grade II gliomas, and grade I gliomas ( Figure 4 In addition, survival analysis showed that high levels of G6PD protein in these 145 glioma tissues were closely associated with low patient survival rates ( Figure 5 These results suggest that G6PD may be a new molecular marker for GBM and may play an important role in the occurrence and development of GBM.
[0063] Example 2: G6PDi enhances the sensitivity of cancer cells to lipid synthesis inhibitors The RNAseq data of 153 primary tumor patients in the GBM cohort in TCGA were analyzed. The results showed that the expression of G6PD was significantly positively correlated with the expression of lipid synthesis-related genes such as SREBP1, FASN, ACC, etc., and was also significantly positively correlated with the expression of genes related to maintaining intracellular redox homeostasis such as GPX4, TXN, B166, etc. ( Figure 6 ).
[0064] To evaluate the potential clinical significance of our observations, we performed a correlation analysis of the immunohistochemical staining patterns of SREBP-1 and G6PD in tumor tissues of 16 GBM patients on tissue microarray (TMA). The results showed that the expression levels of SREBP-1 and G6PD in tumor tissues of GBM patients were significantly positively correlated ( Figure 7 , some cases), among which G6PD expression was detected in 15 GBM tumor samples.
[0065] Example 3: G6PDi combined with inhibitors of new molecular points for the treatment of GBM GBM cells were treated with G6PDi-1 and a novel molecular point inhibitor to test their synergistic therapeutic effect. The drugs were purchased from MedChemExpress.
[0066] Treatment of GBM cells U87 with the SREBP1 inhibitor Fatostatin (2μM) combined with the G6PD inhibitor G6PDi-1 (5μg / ml, 10μg / ml) for 3 days produced a dose-dependent growth inhibition and promoted U87 cell death. However, treatment of U87 with Fatostatin (2μM) or G6PDi-1 (5μg / ml, 10μg / ml) alone for 3 days did not produce or only produced a weak growth inhibition effect ( Figure 8-9 , Fatostatin).
[0067] Treatment of GBM cells U251 with the DGAT1 inhibitor A922500 (DGATi, 20μg / ml) combined with the G6PD inhibitor G6PDi-1 (5μg / ml, 10μg / ml) for 4 days produced a dose-dependent growth inhibition and promoted U251 cell death. However, treatment of U251 with A922500 (20μg / ml) or G6PDi-1 (5μg / ml, 10μg / ml) alone for 4 days did not produce or only produced a weak growth inhibition effect ( Figure 10-11 A922500). After 2 weeks of treatment with the DGAT1 inhibitor A922500 (DGATi, 2μg / ml, 5μg / ml) combined with the G6PD inhibitor G6PDi-1 (5μg / ml, 10μg / ml), GBM cells U251 showed a dose-dependent growth inhibition and promoted U251 cell death ( Fig.12 , A922500 colony formation assay).
[0068] Treatment of GBM cells U251 with the GPX4 inhibitor RSL-3 (2μM) combined with the G6PD inhibitor G6PDi-1 (5μg / ml, 10μg / ml) for 4 days produced a dose-dependent growth inhibition and promoted U251 cell death. However, treatment of U251 with RSL-3 (2μM) or G6PDi-1 (5μg / ml, 10μg / ml) alone for 4 days only produced a weak growth inhibition effect ( Figure 13-14 , RSL-3).
Claims
1. A pharmaceutical composition for treating brain glioma, characterized in that: The pharmaceutical composition comprises: (i) a therapeutically effective amount of a G6PD inhibitor; (ii) a therapeutically effective amount of a SREBP1 inhibitor; Preferably, the G6PD inhibitor is G6PDi-1 or a pharmaceutically acceptable salt thereof; Preferably, the SREBP1 inhibitor is Fatostatin or a pharmaceutically acceptable salt thereof.
2. The pharmaceutical composition according to claim 1, characterized in that The dosage ratio of the G6PD inhibitor to the SREBP1 inhibitor is 1:0.1-2; Preferably, the dosage ratio of the G6PD inhibitor to the SREBP1 inhibitor is selected from 3:1, 2:1, 1:1, 1:2, 1:3; Preferably, the G6PD inhibitor and the SREBP1 inhibitor are present in the same or different compositions or kits; Preferably, the use of the G6PD inhibitor and the SREBP1 inhibitor includes concurrent administration or sequential administration. Specifically, the administration is to provide therapeutically effective levels of the G6PD inhibitor and the SREBP1 inhibitor in vivo.
3. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient. Specifically, the pharmaceutically acceptable carrier or excipient is a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or any type of formulation auxiliary agent.
4. The pharmaceutical composition according to claim 1, characterized in that The brain glioma includes grade I, grade II, grade III, and grade IV brain glioma; Preferably, the brain glioma is a grade IV brain glioma; Preferably, the brain glioma is glioblastoma multiforme.
5. The pharmaceutical composition according to claim 1, characterized in that The dosage forms of the G6PD inhibitor and the SREBP1 inhibitor may be the same or different; Preferably, the dosage form includes drops, tablets, capsules, granules, films, gels, powders, emulsions, pills, suppositories, aerosols, sprays, powder mists, patches, solutions, ointments or creams.
6. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a product for treating brain glioma.
7. The use according to claim 6, characterized in that: The product includes a kit, a chip, a test strip, or a device, an apparatus, or a computer-readable storage medium; Preferably, the product may also contain other drugs for treating cancer; Preferably, the product may also contain other drugs for treating brain glioma; Preferably, the brain glioma includes grade I, grade II, grade III, and grade IV brain glioma; Preferably, the brain glioma is a grade IV brain glioma; Preferably, the brain glioma is glioblastoma multiforme.
8. Use of a reagent for detecting the expression level of G6PD, characterized in that: The application includes any of the following: 1) Application in the preparation of products for diagnosing brain glioma; 2) Application in the preparation of products for grading brain gliomas; 3) Application in the preparation of products for predicting the prognosis of brain glioma.
9. The use according to claim 8, characterized in that: The test is performed on a sample from a patient or subject, and the sample includes tissue, cells, blood, urine, saliva, mucosal samples, feces, intestinal lavage, joint fluid, cerebrospinal fluid, bile samples, respiratory secretions, and bronchoalveolar lavage fluid samples; Preferably, the sample is tissue or cell.
10. The use according to claim 8, characterized in that: The brain glioma includes grade I, grade II, grade III, and grade IV, and the expression of G6PD increases with the grade; Preferably, the brain glioma is a grade IV brain glioma; Preferably, the brain glioma is glioblastoma multiforme; Preferably, the reagent for detecting the expression amount of G6PD includes a reagent for detecting the expression amount of G6PD protein and / or the expression amount of G6PD mRNA; Preferably, the reagents for detecting the expression of G6PD protein include reagents used in the following methods: hematoxylin-eosin staining, safranin O-fast green staining, protein blotting, enzyme-linked immunosorbent assay, radioimmunoassay, sandwich assay, immunohistochemical staining method, mass spectrometry, immunoprecipitation analysis, complement fixation analysis, flow cytometry fluorescence sorting technology and protein chip method; Preferably, the reagents for detecting the expression of G6PD mRNA include reagents used in the following methods: PCR-based detection method, Southern hybridization method, Northern hybridization method, dot hybridization method, fluorescence in situ hybridization method, DNA microarray method, ASO method, and high-throughput sequencing platform method.