Application of combination of glibenclamide and halofuginone in preparation of preparation for preventing and / or treating radiation-induced lung injury
By combining glibenclamide and styraxone, a multi-target intervention network is formed to synergistically block the pathological process of radiation-induced lung injury, solving the problems of large side effects and poor efficacy of existing drugs, and achieving highly efficient treatment and prevention of radiation-induced lung injury.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-25
- Publication Date
- 2026-03-10
AI Technical Summary
Existing drugs for treating radiation-induced lung injury have significant side effects and poor therapeutic efficacy, making it difficult to effectively block the progression of pulmonary fibrosis.
Glibenclamide and styraxone are used together in a mass ratio of 1:5 to 15 to form a drug composition. By regulating the energy metabolism of lung tissue cells, inhibiting fibroblast senescence and directly inhibiting the TGF-β signaling pathway, they synergistically exert anti-fibrotic, anti-inflammatory and antioxidant effects, thus blocking the pathological process of radiation-induced lung injury.
It is significantly superior to single-drug intervention, reduces the degree of pathological damage, improves disease prognosis, and is suitable for the early prevention of acute radiation pneumonitis and the long-term treatment of radiation pulmonary fibrosis, while maintaining good medication safety.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological medicine, and particularly relates to application of glibenclamide combined with halofuginone in preparation of preparations for preventing and / or treating radiation-induced lung injury. BACKGROUND
[0002] Radiation-Induced Lung Injury (RILI) is one of the most common and serious complications in the process of radiotherapy for chest tumors (such as lung cancer, esophageal cancer, breast cancer, etc.), and its mechanism is complex, mainly involving direct damage of radiation to lung tissue cells, oxidative stress response, inflammatory response and subsequent lung fibrosis process. Clinically, radiation-induced lung injury is usually divided into two stages: acute radiation pneumonia and chronic radiation pulmonary fibrosis. The acute stage usually occurs several weeks to several months after radiotherapy, and is characterized by symptoms such as cough, shortness of breath and fever. The chronic stage occurs several months to several years after radiotherapy, and is mainly characterized by lung tissue fibrosis and progressive decline in lung function, which seriously affects the quality of life and survival of patients.
[0003] At present, the treatment means for radiation-induced lung injury is limited, and the commonly used drugs in clinic include glucocorticoids, antioxidants (such as ambroxol, N-acetylcysteine) and anti-inflammatory drugs. However, long-term use of glucocorticoids will bring a series of side effects such as immune suppression, increased risk of infection and elevated blood sugar; the therapeutic effect of existing antioxidants and anti-inflammatory drugs is often not ideal, and it is difficult to effectively block the progression of pulmonary fibrosis. Therefore, it is of great clinical significance and application value to find safe and efficient drugs or drug combinations for preventing and treating radiation-induced lung injury. SUMMARY
[0004] The present application aims to provide application of glibenclamide combined with halofuginone in preparation of preparations for preventing and / or treating radiation-induced lung injury.
[0005] In order to achieve the above-mentioned application purposes, the present application provides the following technical solutions: The present application provides application of glibenclamide combined with halofuginone in preparation of preparations for preventing and / or treating radiation-induced lung injury.
[0006] Preferably, the mass ratio of glibenclamide to halofuginone is 1:5-15.
[0007] Preferably, the radiation-induced lung injury is lung injury caused by ionizing radiation.
[0008] Preferably, the ionizing radiation includes X-ray radiation, gamma-ray radiation or heavy ion radiation.
[0009] Preferably, the radiation-induced lung injury includes acute radiation pneumonia or radiation-induced pulmonary fibrosis.
[0010] The present application also provides a pharmaceutical composition for preventing and / or treating radiation-induced lung injury, which comprises glibenclamide, halofuginone and a pharmaceutically acceptable carrier thereof.
[0011] Preferably, the mass ratio of glibenclamide to halofuginone is 1:5-15.
[0012] The present application also provides the use of the pharmaceutical composition in the preparation of a preparation for preventing and / or treating radiation-induced lung injury.
[0013] The present application also provides the use of the pharmaceutical composition in the preparation of a preparation for preventing and / or treating acute radiation pneumonitis.
[0014] The present application also provides the use of the pharmaceutical composition in the preparation of a preparation for preventing and / or treating radiation-induced lung fibrosis.
[0015] The present application has the following advantages: The present application provides a pharmaceutical composition formed by combining glibenclamide and halofuginone at a mass ratio of 1:5-15, which exhibits significant synergistic technical advantages in preventing and treating radiation-induced lung injury caused by ionizing radiation, and provides an efficient new strategy for clinical intervention of the disease. The core technical effect is derived from the complementation and synergistic effect of the action mechanisms of the two drugs, and through the construction of a multi-target and all-round intervention network, the precise blockage of the pathological process of radiation-induced lung injury is achieved. Glibenclamide can regulate lung tissue cell energy metabolism, inhibit fibroblast aging, and indirectly regulate the intensity of inflammatory response. Halofuginone can directly inhibit the TGF-β signaling pathway, block fibroblast activation and collagen deposition, and at the same time, remove oxidative stress products and reduce the release of pro-inflammatory factors, thereby playing a clear anti-fibrosis, anti-inflammatory and anti-oxidation role. When the two drugs are used in combination, a synergistic effect of "inflammation inhibition-fibrosis block-cell protection" is formed, which not only strengthens the control of early acute inflammation, but also effectively delays the progression of later lung fibrosis. It plays a protective role from the source of pathological damage to the whole process of progression, and is significantly superior to the intervention effect of a single drug. Moreover, the pharmaceutical composition has a wide range of applications, which can not only meet the early prevention and emergency intervention needs of acute radiation pneumonitis, but also adapt to the long-term treatment scenario of radiation-induced lung fibrosis. Based on the existing clinical application of the two drugs, the combination still maintains good drug safety and tolerance.
[0016] Therefore, the pharmaceutical composition of glibenclamide and halofuginone breaks through the limitations of single drug intervention through mechanism synergy, effectively reduces the incidence risk of radiation-induced lung injury, reduces the degree of pathological damage, and improves the prognosis of the disease, thereby providing a new technical path for the prevention and treatment of radiation therapy-related lung injury, and having important clinical translation value and application prospect. BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 Glibenclamide combined with halofuginone on RLE-6TN cell line cytotoxicity; Figure 2 is the lung tissue HE staining results of each group of mice on the 20th day after ionizing radiation; Figure 3 is the survival curve of the mouse model of radiation-induced lung injury. DETAILED DESCRIPTION
[0018] The application provides application of glibenclamide combined with halofuginone in preparation of preparations for preventing and / or treating radiation-induced lung injury.
[0019] In the application, the glibenclamide is an organic compound with a chemical formula of C 23 H 28 CIN3O5S, which is a sulfonylurea hypoglycemic drug, is suitable for mild and moderate type Ⅱ diabetes who is not satisfied with diet control alone, has a certain insulin secretion function of pancreatic beta cells, and has no serious complications; the halofuginone is also known as halofuginone hydrobromide, with a chemical name of 7-bromo-6-chloro-3-[3-(3-hydroxy-2-piperidyl)-2-oxopropyl (propionyl)]-4(3H)-quinazolinone hydrobromide, which is a quinazolinone drug with anti-coccidiosis extracted from traditional Chinese medicine halofuginone; The mass ratio of the glibenclamide to the halofuginone is 1:5-15, preferably 1:8-12, and more preferably 1:10.
[0020] In the application, the radiation-induced lung injury is lung injury caused by ionizing radiation.
[0021] In the application, the ionizing radiation includes X-ray radiation, γ-ray radiation or heavy ion radiation.
[0022] In the application, the radiation-induced lung injury includes acute radiation pneumonia or radiation-induced pulmonary fibrosis.
[0023] The application further provides a pharmaceutical composition for preventing and / or treating radiation-induced lung injury, which comprises glibenclamide, halofuginone and a pharmaceutically acceptable carrier thereof.
[0024] In the application, the mass ratio of the glibenclamide to the halofuginone is 1:5-15, preferably 1:8-12, and more preferably 1:10.
[0025] The application further provides application of the pharmaceutical composition in preparation of preparations for preventing and / or treating radiation-induced lung injury.
[0026] The application further provides application of the pharmaceutical composition in preparation of preparations for preventing and / or treating acute radiation pneumonia.
[0027] The present invention also provides the use of the pharmaceutical composition described herein in the preparation of agents for the prevention and / or treatment of radiation-induced pulmonary fibrosis.
[0028] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0029] In the embodiments of the present invention, the glibenclamide was purchased from Shenzhen Feisi Biotechnology Co., Ltd., fentanyl was purchased from Xi'an Sinote Biotechnology Co., Ltd., amifostine was purchased from Hubei Jianyuan Biochemical Co., Ltd., CCK-8 reagent was purchased from Dojin Chemical Research Institute of Japan, and TUNEL assay kit was purchased from Yisheng Biotechnology (Shanghai) Co., Ltd. In the test materials of this invention, the test mice were 6-8 week old C57BL / 6 strain mice, weighing 18-22g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.; the test cell line was rat type II alveolar epithelial cell RLE-6TN cell line (catalog number: Delf-16735), purchased from Hefei Wanwu Biotechnology Co., Ltd.
[0030] Example 1: Cytotoxicity of glibenclamide and styraxone
[0031] RLE-6TN cell lines were seeded in DMFM / F12 medium containing 10% fetal bovine serum and cultured at 37°C and 5% CO2 until the logarithmic growth phase. The cells were then seeded into 96-well plates and divided into glibenclamide, styraxone, and control groups. After 24 hours of further culture, glibenclamide was added to the wells of the glibenclamide groups to final concentrations of 6.25 μmol / L, 12.5 μmol / L, 25 μmol / L, 50 μmol / L, and 100 μmol / L, respectively. The concentrations of styraxone were increased to 6.25 μmol / L, 12.5 μmol / L, 25 μmol / L, 50 μmol / L, 100 μmol / L, and 200 μmol / L, respectively, in the culture wells of the styraxone group. No reagent was added to the control group. After culturing for 48 h, CCK-8 reagent was added to each well according to the instructions and culturing for 2 h. The OD values of each well were then measured using a microplate reader. 450 The nm value was calculated and cell viability was determined. The results are as follows: Figure 1 As shown, the formula for calculating cell viability is: Cell viability (%) = .
[0032] The results showed that glibenclamide at concentrations ≤100 μmol / L and styraxone at concentrations ≤50 μmol / L had no significant toxicity to type II alveolar epithelial cells of RLE-6TN rats (survival rate ≥90%), and this concentration range provided a safe basis for subsequent combination therapy.
[0033] Example 2: The preventive and therapeutic effects of glibenclamide combined with styraxone on radiation-induced lung injury.
[0034] 180 C57BL / 6 mice were divided into 9 groups: blank control group, irradiation group, glibenclamide group, styraxone group, low-dose combination group, medium-dose combination group, high-dose combination group, negative control group, and positive control group, with 20 mice in each group. One hour before ionizing radiation, mice in the glibenclamide group were injected intraperitoneally with 0.2 mg / kg glibenclamide, mice in the styraxone group were injected intraperitoneally with 1.0 mg / kg styraxone, mice in the low-dose combination group were injected intraperitoneally with 0.1 mg / kg glibenclamide + 0.5 mg / kg styraxone, mice in the medium-dose combination group were injected intraperitoneally with 0.2 mg / kg glibenclamide + 1.0 mg / kg styraxone, mice in the high-dose combination group were injected intraperitoneally with 0.2 mg / kg glibenclamide + 2.0 mg / kg styraxone, mice in the blank control group were injected intraperitoneally with 0.1 mL of physiological saline, mice in the irradiation-only group and the negative control group were injected intraperitoneally with 0.1 mL of dimethyl sulfoxide, and mice in the positive control group were injected intraperitoneally with 20 mg / kg amifostine. Mice in the simple irradiation group, glibenclamide group, styraxone group, low-dose combination group, medium-dose combination group, high-dose combination group, and positive control group were anesthetized by intraperitoneal injection of 50 mg / kg 1% sodium pentobarbital solution and fixed in a supine position. The lungs of each mouse were subjected to γ-ray ionizing radiation using 60Co as the radiation source at a dose of 30 Gy and a radiation rate of 1 Gy / min. Lead plates were used to shield other parts of the mice to prevent ionizing radiation damage. The blank control group and negative control group mice were not subjected to ionizing radiation. After ionizing radiation, bilateral lungs were harvested on days 7, 14, and 20, fixed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE). Figure 2 The lungs of each mouse were washed with pre-cooled physiological saline, and the fresh wet weight and the dry weight after drying were weighed. The wet / dry ratio and lung coefficient were calculated according to the formula. The results are shown in Table 1. The apoptosis rate of vascular endothelial cells in the lungs of each mouse was determined by the Tunel assay kit. The results are shown in Table 2. For specific steps, please refer to the instructions for use of the Tunel assay kit. Wet / dry density = Fresh wet weight of left lung / Dry weight of left lung after drying; Lung coefficient (%) = Lung wet weight (g) / Body weight (g) × 100%; Table 1. Wet / dry weight and lung coefficient of mice in each group.
[0035] Table 2 Apoptosis rate of vascular endothelial cells in each group of mice
[0036] As shown in Table 1, mice in the irradiation-only group exhibited severe pulmonary edema on days 7, 14, and 20, with significantly higher lung wet / dry ratios and lung coefficients compared to the control group, indicating successful model establishment. Meanwhile, the TUNEL assay results in Table 2 showed a sharp increase in the apoptosis rate of pulmonary vascular endothelial cells in the irradiation-only group to 88.35%, suggesting that apoptosis is a key mechanism of radiation damage. Pretreatment with the positive control drug amifostine (20 mg / kg) significantly reduced pulmonary edema and inhibited apoptosis, confirming the model's effectiveness and evaluation criteria. While monotherapy with glibenclamide (0.2 mg / kg) or styraxone (1.0 mg / kg) could alleviate pulmonary edema and apoptosis to some extent, their effects were weaker than those of amifostine. Amifostine; however, the combination of the two drugs showed a significant synergistic effect. The low-dose combination group (0.1 mg / kg glibenclamide + 0.5 mg / kg fentanyl) was more effective than either drug but less effective than the positive control. More importantly, the medium-dose combination group (0.2 mg / kg glibenclamide + 1 mg / kg fentanyl) showed the strongest protective effect among all treatment groups: its lung wet / dry ratio (1.90) and lung coefficient (0.88%) on day 20 were lower than those of the positive control group (2.40 and 1.02%), and its vascular endothelial cell apoptosis rate (16.85%) was also significantly lower than that of the positive control group (35.87%). The high-dose combination group showed similar effects to the medium-dose group, without further enhancement. Therefore, the combination of glibenclamide and fentanyl, especially the medium-dose regimen, showed superior effects to the classic protective agent amifostine in both reducing radiation-induced pulmonary edema and inhibiting pulmonary vascular endothelial cell apoptosis. Its protective effect may be due to the synergistic inhibition of key radiation-induced cell death pathways by the two drugs. from Figure 2As can be seen, the lung tissue structure of the blank control group mice was normal, the alveolar walls were thin and elastic, the alveolar cavities were round and evenly distributed, and there were no pathological features such as inflammatory reactions and hemorrhage, consistent with the negative control group. After 20 days of ionizing radiation, the lung tissue capillaries of the mice in the simple irradiation group were significantly dilated and congested, and red blood cells leaked into the alveolar cavities due to damage to the blood vessel walls. Local alveolar cavities collapsed, atelectasised or merged into bullae, and various inflammatory cell infiltrations appeared in the interstitium. The alveolar walls were significantly widened, edematous, compressed, and adhered, and partial alveolar cavities were also observed. The presence of large-scale fusion, severe intracavitary hemorrhage with fibrinous exudate, and brownish-red refractory granules formed after macrophages engulfed excessive red blood cells, fully demonstrates the successful modeling of radiation-induced lung injury. The lung tissue lesions in the positive control group (amifostine), glibenclamide group, and thiazoline group were comparable, all exhibiting some degree of interstitial edema, alveolar wall widening and fusion, and alveolar congestion. However, compared to the irradiation-only group, the overall inflammation and damage were milder, and some alveolar structures and outlines were still discernible, indicating... All three intervention methods, used individually, could prevent and alleviate radiation-induced lung injury to a certain extent, and there was no significant difference in the preventive and therapeutic effects of glibenclamide and styraxone alone. Although mice in the low-dose combination group (glibenclamide + styraxone) still exhibited interstitial pulmonary edema and varying degrees of alveolar wall fusion, the overall degree of inflammation was lower than that in the irradiation-only group, the positive control group, the glibenclamide group, and the styraxone group. The degree of lesions in alveolar contour and structure was also significantly reduced, initially demonstrating the synergistic anti-radiation and anti-inflammatory effects of glibenclamide and styraxone. The medium-dose combination... In the lung tissue of the mice, most alveolar cavities had clear structures and outlines, alveolar wall thickening was not obvious, pulmonary interstitial edema was mild, and there was no obvious fusion or adhesion between most alveolar walls. Although they did not fully reach the healthy levels of the blank control group and the negative control group, the degree of alveolar inflammation and alveolar wall thickness were significantly better than those of the single-drug group and the positive control group, and the staining results were close to those of the high-dose combination group. This indicates that when glibenclamide and styraxone are used in combination at medium and high doses, they can show significant radiation-induced lung injury prevention and treatment effects without dose dependence.
[0037] Example 3: Effects of glibenclamide combined with styraxone on TNF-α and TGF-β1 levels
[0038] Many studies have found that transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) are involved in the early inflammatory response of radiation-induced lung injury and can also affect the later fibrosis process by regulating the proliferation, differentiation and collagen fiber deposition of fibroblasts. Blood samples from mice in each group described in Example 2 were taken, and the contents of TNF-α (ng / L) and TGF-β1 (ng / L) in plasma were detected by ELISA method according to the literature (Zhou Yanping, Qiu Mingyi, Hu Zuowei, et al. Effects of Sha Shen Mai Dong Tang on plasma IL-6, TNF-α and TGF-β1 in radiation-induced pneumonia rats [J]. Chinese Journal of Experimental Traditional Medical Formulae, 2014.). The results are shown in Table 3. Table 3. Expression levels of TNF-α and TGF-β1 in mice of each group
[0039] Table 3 shows that ionizing radiation significantly upregulated the levels of key inflammatory cytokines TNF-α and the pro-fibrotic factor TGF-β1 in mouse plasma. The plasma TNF-α and TGF-β1 levels in the irradiation-only group were significantly higher than those in the blank control and negative control groups at all time points. TNF-α levels peaked on day 14 after irradiation, while TGF-β1 levels continued to increase over time, consistent with its role in driving chronic fibrosis. Pretreatment with the positive control drug amifostine effectively inhibited the release of these cytokines. Compared with the irradiation-only group, the plasma TNF-α and TGF-β1 levels in the amifostine group were significantly lower at all time points, confirming the effectiveness of the model. Glibenclamide or... Xanthine monotherapy showed limited inhibitory effects; the inhibitory effects of glibenclamide and xanthine on the two cytokines were weaker than those in the positive control group; the combination of glibenclamide and xanthine showed a significant synergistic inhibitory effect, with the low-dose combination group showing an effect comparable to the positive control group; more importantly, the medium-dose combination group (0.2 mg / kg glibenclamide + 1.0 mg / kg xanthine) and the high-dose combination group showed the most significant inhibitory effects on TNF-α and TGF-β1, with plasma levels lower than those in the positive control group at all time points; therefore, the combination of glibenclamide and xanthine can synergistically inhibit key early inflammatory and pro-fibrotic signals in radiation-induced lung injury, and its effect is superior to that of the classic radioprotective agent amifostine.
[0040] Example 4: Establishment of a mouse model of radiation-induced lung injury
[0041] C57BL / 6 mice were anesthetized with 50 mg / kg 1% sodium pentobarbital solution via intraperitoneal injection and fixed in a supine position. Local gamma-ray ionizing radiation was applied to the chest of the C57BL / 6 mice using 60Co as the radiation source at a dose of 30 Gy and a radiation rate of 200 cGy / min. Lead plates were used to shield other parts of the C57BL / 6 mice to prevent ionizing radiation damage. Following ionizing radiation, the C57BL / 6 mice exhibited acute radiation-induced pneumonia within 1–2 weeks, and bilateral lung fibrosis symptoms appeared within 3–5 weeks, largely consistent with the course of radiation-induced lung injury in humans, indicating successful establishment of a mouse model of radiation-induced lung injury.
[0042] Example 5: Effect of glibenclamide combined with styraxone on survival rate of mice with radiation-induced lung injury.
[0043] One hundred mice with radiation-induced lung injury, constructed in Example 4, were randomly divided into five groups: irradiation group, glibenclamide group, styraxone group, combination therapy group, and positive control group, with 20 mice in each group. Six hours after ionizing radiation, mice in the glibenclamide group were intraperitoneally injected with 0.2 mg / kg glibenclamide, mice in the styraxone group were intraperitoneally injected with 1.0 mg / kg ferulic acid, mice in the combination therapy group were intraperitoneally injected with 0.2 mg / kg glibenclamide + 1.0 mg / kg styraxone, and mice in the positive control group were intraperitoneally injected with 20 mg / kg amifostine. The corresponding medications were administered every two days thereafter. Mice in the irradiation group received no medication. The survival rate of mice in each group was recorded over 20 days. The results are as follows: Figure 3 As shown; from Figure 3 As can be seen, during the 20-day observation period, the survival rate of mice in the irradiated group who did not receive any treatment dropped sharply, with a final survival rate of only about 10%, indicating that the model was successfully established and the damage was severe. Each drug treatment group showed different degrees of protective effect. Among them, the glibenclamide group and the styraxone monotherapy group could delay the death process and improve the final survival rate, but their effects were limited. The positive control drug amifostine group showed a clear protective effect, and the survival rate curve was significantly higher than that of the irradiated group. However, the survival curve of the glibenclamide and styraxone combination drug group was always at the top, and its survival rate remained the highest and the decline trend was the most gradual throughout the observation period, which was significantly better than all other groups at the experimental endpoint. Therefore, the combination of glibenclamide and styraxone has a significant synergistic effect and is better than single drug treatment and classic positive control drugs in improving the survival prognosis of animal models of radiation lung injury.
[0044] As can be seen from the above embodiments, the present invention provides the application of glibenclamide combined with styraxone in the preparation of formulations for the prevention and / or treatment of radiation-induced lung injury.
[0045] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. Use of glibenclamide combined with halofuginone in the preparation of a preparation for preventing and / or treating radiation-induced lung injury.
2. Use according to claim 1, characterized in that, The mass ratio of the glibenclamide to the halofuginone is 1:5-15.
3. Use according to claim 1, characterized in that, The radiation-induced lung injury is ionizing radiation-induced lung injury.
4. Use according to claim 3, characterized in that, The ionizing radiation includes X-ray radiation, γ-ray radiation or heavy ion radiation.
5. The use according to claim 1, characterized in that, The radiation-induced lung injury includes acute radiation pneumonitis or radiation-induced pulmonary fibrosis.
6. A pharmaceutical composition for use in the prevention and / or treatment of radiation-induced lung injury, characterized in that, The pharmaceutical composition comprises glibenclamide, halofuginone and a pharmaceutically acceptable carrier thereof.
7. The pharmaceutical composition of claim 6, wherein, The mass ratio of the glibenclamide to the halofuginone is 1:5-15.
8. Use of the pharmaceutical composition of claim 6 or 7 in the preparation of a preparation for preventing and / or treating radiation-induced lung injury.
9. Use of the pharmaceutical composition of claim 6 or 7 in the preparation of a preparation for preventing and / or treating acute radiation pneumonitis.
10. Use of the pharmaceutical composition of claim 6 or 7 in the preparation of a preparation for preventing and / or treating radiation-induced pulmonary fibrosis.