Application of flavonoid compound 5-demethylated sweet orange flavone in preparation of medicine for treating hemogenic acute radiation syndrome

Abstract

The invention discloses an application of a flavonoid compound 5-demethylated sweet orange flavone in preparation of a medicine for treating hemogenic acute radiation syndrome, and relates to the technical field of biological medicines. The medicine 5-demethylated sweet orange flavone (DMSS) is obtained through high-throughput screening of a zebra fish irradiation granulocyte reduction (granulocyte reduction) model, and the new application of the medicine for effective radiation resistance is verified in cell and mouse irradiation models. The invention discloses the protective effect of DMSS in irradiation-induced hematopoietic injury for the first time, irradiation protection is realized by relieving irradiation-induced neutrophil apoptosis, and as a novel protective agent, the anti-oxidation, anti-inflammatory, anti-tumor and other activities of DMSS are proved in various disease models. Therefore, compared with a synthetic radiation protective agent, DMSS has lower toxic and side effects and better clinical transformation potential, and the unique action mechanism of DMSS provides a new candidate molecule for drug treatment of acute radiation injury of a hematopoietic system.

Description

TECHNICAL FIELD

[0001] The application relates to the field of biological medicine, and in particular to application of flavonoid compound 5-demethyllauchingerin in preparation of a drug for treating hematopoietic acute radiation syndrome. BACKGROUND

[0002] Acute radiation syndrome (ARS) is caused by the exposure of the body to penetrating radiation, which can damage bone marrow cells and further cause hematopoietic acute radiation syndrome (H-ARS), the clinical features of which include neutropenia, thrombocytopenia and anemia, and which is potentially fatal. Given the high sensitivity of hematopoietic tissues to ionizing radiation, accurate diagnosis and effective treatment of H-ARS are crucial for the prognosis of individuals exposed to radiation.

[0003] However, the US Food and Drug Administration (FDA) has only approved seven drugs for the treatment of H-ARS, including five drugs with human granulocyte colony-stimulating factor (G-CSF) as the core scaffold: Neupogen, Neulasta, Releuko, Fylnetra, Zarxio, and two other drugs: Leukine (recombinant human granulocyte-macrophage colony-stimulating factor, GM-CSF) and Nplate (thrombopoietin receptor agonist). As can be seen, promoting the production and survival of neutrophils is an important treatment strategy for H-ARS. However, it is worth noting that the existing drugs for H-ARS are limited in type, and all show certain adverse reactions in clinical trials, such as G-CSF, which often causes mild to moderate bone pain, diarrhea, fever and skin rash, etc. while improving neutropenia. Therefore, the development of new radiation protection drugs has important clinical urgency. In addition, compared with protein drugs, small molecule drugs have the advantages of oral availability, high stability, strong tissue penetration, targeting of intracellular molecules, low synthesis and storage cost, and low risk of immunogenicity. Therefore, the development of anti-radiation small molecule drugs may have important scientific and application value. SUMMARY

[0004] The purpose of the present application is to solve the above problems, and to provide application of flavonoid compound 5-demethyllauchingerin in preparation of a drug for treating hematopoietic acute radiation syndrome. The drug 5-demethyllauchingerin (DMSS) obtained through high-throughput screening of a zebrafish irradiation granulocytopenia (granulocytopenia) model, and its new use as an effective anti-radiation drug by reducing radiation-induced neutrophil apoptosis is verified in cell and mouse irradiation models.

[0005] The application discloses application of a flavonoid compound, 5-demethylloniflavone, in preparation of a medicine for treating hematopoietic acute radiation syndrome.

[0006] Preferably, in the application technical scheme, the 5-demethylloniflavone realizes the irradiation protection effect by affecting cell apoptosis.

[0007] Preferably, in the application technical scheme, the 5-demethylloniflavone realizes the irradiation protection effect by reducing irradiation-induced neutrophil apoptosis.

[0008] Preferably, in the zebrafish irradiation granulocyte reduction model, the 5-demethylloniflavone realizes the irradiation protection effect by alleviating irradiation-induced reduction of the number of neutrophils, and the ability of the 5-demethylloniflavone to restore the number of neutrophils is concentration-dependent.

[0009] Preferably, in the cell irradiation model, the 5-demethylloniflavone realizes the protection effect on neutrophils by increasing the expression of anti-apoptotic protein BCL-2 and reducing the expression of pro-apoptotic protein BAX.

[0010] Preferably, in the mouse irradiation model, the 5-demethylloniflavone has at least one of the following effects:

[0011] (1) reducing or alleviating the clinical symptoms of hematopoietic acute radiation syndrome;

[0012] (2) improving the survival rate;

[0013] (3) reducing irradiation-induced apoptosis of the number of peripheral blood neutrophils;

[0014] (4) reducing irradiation-induced apoptosis of the number of splenic myeloid cells.

[0015] Preferably, in the application technical scheme, the medicine is a pre-irradiation exposure medicine.

[0016] Preferably, in the application technical scheme, the dosage form of the medicine is a hard capsule, a soft capsule, a powder, a granule, a tablet, a pill, a honeyed paste, an oral liquid, a suppository, a liquor, or an injection.

[0017] Preferably, in the application technical scheme, the medicine further comprises a pharmaceutically acceptable carrier.

[0018] To sum up, the present application includes at least one of the following beneficial technical effects:

[0019] 5-Desmethylsinensetin (DMSS) belongs to flavonoids of natural origin, which has excellent biocompatibility and safety basis. The present application first discloses the protective effect of DMSS on hematopoietic injury caused by irradiation. Through zebrafish irradiation model, cell irradiation model and mouse irradiation model, it is determined that DMSS realizes irradiation protection by relieving neutrophil apoptosis caused by irradiation. As a new protective agent, the antioxidant, anti-inflammatory and antitumor activities of DMSS have been confirmed in various disease models. Therefore, compared with synthetic radiation protectants, DMSS has lower toxic side effects and better clinical transformation potential. Its unique mechanism provides a new candidate molecule for drug treatment of acute radiation damage of hematopoietic system. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is a zebrafish model screening result graph, wherein Figure 1 A is a drug screening process schematic diagram, Figure 1 B is a representative image of 96hpf whole body irradiated zebrafish during screening, Figure 1 C is a drug screening result schematic diagram, Figure 1 D is a result schematic diagram of DMSS restoring the number of neutrophils in the zebrafish irradiated granulocyte reduction model;

[0021] Figure 2 is a cell test result graph, wherein Figure 2 A is a result schematic diagram of the effect of irradiation dose on cell viability, Figure 2 B is a result schematic diagram of the effect of different concentrations of DMSS administration on cell viability, Figure 2 C and Figure 2 D is a flow cytometry result schematic diagram of cell apoptosis detection, Figure 2 E is a protein band diagram of WB experiment, Figure 2 F is a diagram of the relationship between BAX protein expression level and DMSS concentration, Figure 2 G is a diagram of the relationship between BCL-2 protein expression level and DMSS concentration;

[0022] Figure 3 is a mouse test result graph, wherein Figure 3 A is a mouse irradiation test flow chart, Figure 3 B is a mouse survival result schematic diagram, Figure 3 C is a mouse weight analysis schematic diagram, Figure 3 D is a mouse blood routine test result schematic diagram, Figure 3 E and Figure 3 F is a mouse spleen myeloid cell detection result schematic diagram;

[0023] Figure 4 is a statistical diagram of the effect of DMSS in zebrafish model, cell model and mouse model, whereinFigure 4 A and Figure 4 B is the effect diagram of DMSS in the zebrafish irradiation granulocytopenia model, Figure 4 C and Figure 4 D is the effect diagram of DMSS in the cell test, Figure 4 E and Figure 4 F is the effect diagram of DMSS in the mouse test. DETAILED DESCRIPTION

[0024] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below in combination with embodiments and drawings. It should be understood that the specific embodiments described herein are only used to explain the present application and should not be used to limit the present application. Any modification or equivalent replacement made by those skilled in the art on the basis of understanding the technical solutions of the present application without departing from the spirit and scope of the technical solutions of the present application should be covered within the protection scope of the present application.

[0025] The information of test animals, reagents, instruments and equipment used in the following embodiments is as follows: all reagents and chemicals are used as they are without further treatment, and the specific conditions not mentioned are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments not mentioned by the manufacturer are conventional products that can be purchased on the market.

[0026] The research method of the present application: through high-throughput screening of the zebrafish irradiation-induced neutropenia model, flavonoid small molecule compound 5-demethylhesperetin (DMSS) is obtained, and its radiation protection effect is verified in cell and mouse tests.

[0027] 1. Test animals

[0028] All test operations follow the Chongqing Medical University Test Animal Management Standard and the Chongqing Medical University Animal Test Welfare and Ethics Committee Management Charter. All test operations follow the "3R" principle to maximize the welfare of test animals.

[0029] (1) The transgenic zebrafish line Tg(mpx:GFP) is bred, preserved and propagated under unified standard conditions, and the temperature, pH and conductivity of the circulating water are 28±2℃, 7.2-7.4 and 500-550 μs / cm, respectively; the artificial light cycle of the zebrafish room is controlled in cycles of 14 hours of light (8:00-22:00) / 10 hours of darkness (22:00-8:00).

[0030] (2) Female C57BL / 6J mice (20±2 g; 6-8 weeks) are purchased from Hunan Slaik Jingda Test Animal Co., Ltd.; the animals are bred under standard conditions (26±1°C, 50-60% humidity, 12h light-dark cycle) and have free access to food and water.

[0031] 2. Test cells

[0032] HL-60 human myeloid leukemia cells were purchased from National Center for Cell Science (Shanghai, China); the cell line was cultured in RPMI-1640 medium (Solebo, China), with the addition of 10% fetal bovine serum FBS and 1% penicillin / streptomycin; all cells were placed in a constant temperature incubator at 37°C, 5% CO2; the cell lines used for the test were completed within 10 passages.

[0033] 3. Main reagents

[0034] Pronase (10165921001, Roche, Switzerland); Batilol (T3004, TaoShu, China); Dimethyl sulfoxide (DMSO) (D8371, Solebo, China); Tricaine (A5040, Sigma-Aldrich, USA); Drug library (HY-L068, MedChemExpress, USA); 5-demethyl tangeretin (HY-N7632, MedChemExpress, USA); CCK8 reagent (CA1210, Solebo, China); other abbreviations not specified are based on industry-wide common understanding; the source of reagents not specified is purchased through conventional channels.

[0035] I. Examples

[0036] Example 1

[0037] This example establishes a zebrafish model of radiation-induced neutropenia and demonstrates that the animal model can be used for large-scale initial screening of anti-radiation drugs. Based on this, drugs that can effectively restore the number of neutrophils in zebrafish are screened and their protective ability is detected.

[0038] 1.1 Zebrafish model drug screening

[0039] Reference Figure 1 A. Zebrafish embryos 24 hours post fertilization (24hpf) were treated with 50 mg / mL pronase for 10-15 minutes to remove the embryonic chorion. Embryos were arranged in a 96-well plate with 200 μL / well of E3 medium (5 embryos per well), and 100 μL of liquid was removed from each well. Different drugs at 10 μM were prepared and added to the wells; at the same time, Batilol was used as a positive drug to verify the feasibility of the model, i.e., 0.1% DMSO and 1 μM Batilol were used for treatment.

[0040] After 1h treatment, the 96-well plate containing embryos was put into the irradiator (RS 2000pro-225 X-ray irradiator) for 15 Gy X-ray (dose rate 2.046 Gy / min) irradiation; then the embryos were incubated at 28.5℃ to 96 hpf, and then GFP-positive neutrophil fluorescence imaging was performed using a stereomicroscope, cell counting was performed, finally, the drug for further study was determined, (statistical method: one-way ANOVA, compared with the DMSO group, *P<0.05, ****P<0.0001).

[0041] The test results are shown in Figure 1 B, Figure 1 C, in which TBI refers to 96hpf total body irradiation at the time of screening. As can be seen from the figure, by high-throughput screening of the flavonoid small molecule compound library, a drug that can effectively restore the number of neutrophils in zebrafish is successfully screened: 5-demethyl sweet orange flavone (DMSS), and 5-demethyl sweet orange flavone (DMSS) can significantly alleviate the decrease in the number of neutrophils caused by irradiation.

[0042] 1.2 Detection of DMSS irradiation protection ability in zebrafish model

[0043] The difference from 1.1 is that only DMSS is used as the drug, and 0 μM, 1 μM, 2 μM, 5 μM, and 10 μM concentrations are added, respectively, and batilol is used as a positive drug to verify the feasibility of the model, that is, 0.1% dimethyl sulfoxide (DMSO) and 1 μM batilol are used for treatment, respectively.

[0044] After 1h treatment, the 96-well plate containing embryos was put into the irradiator (RS 2000pro-225 X-ray irradiator) for 15 Gy X-ray (dose rate 2.046 Gy / min) irradiation; then the embryos were incubated at 28.5℃ to 96 hpf, and then GFP-positive neutrophil fluorescence imaging was performed using a stereomicroscope, cell counting was performed, finally, the drug for further study was determined, (statistical method: one-way ANOVA, compared with the DMSO group, *P<0.05, ****P<0.0001).

[0045] The results are shown in Figure 1 D, as can be seen from the figure, in the zebrafish irradiation granulocyte reduction model, the protective effect of 5-demethyl sweet orange flavone (DMSS) on neutrophils has a certain concentration dependence.

[0046] Example 2

[0047] The present embodiment carries out cell test on DMSS, and respectively carries out cell irradiation and activity detection and cell apoptosis detection: first, the CCK8 method is used to detect the number change of the irradiated neutrophils after administration; second, the 7-AAD / Annexin V flow detection technology is used to detect the apoptosis change of the irradiated neutrophils after administration, and the Western blot experiment method is used to detect the apoptosis related protein change in the irradiated neutrophils after administration.

[0048] 2.1 Cell irradiation and activity detection

[0049] HL-60 cells were inoculated in 60mm culture dishes at a density of 2x10 6 / mL, and were irradiated with 0, 1, 2, 5, 10 Gy X-rays (dose rate 2.046 Gy / min). After irradiation, incubate for 24h, then perform cell counting (statistical method: one-way ANOVA, compared with IR group, **P<0.01, ****P<0.0001).

[0050] The results are shown in Figure 2 A, as shown in the figure, with the increase of irradiation dose, the number of HL-60 cells gradually decreases, and finally the irradiation dose of 5 Gy which can effectively reduce the number of neutrophils without completely reducing is selected for subsequent experiments.

[0051] HL-60 cells were inoculated in 60mm culture dishes at a density of 2x10 6 / mL. The drug DMSS was dissolved in 0.1% DMSO, then added to the culture medium and diluted to 0.5, 1, 2, 5, 10 μM. After 1h, irradiate with 5Gy X-rays (dose rate 2.046 Gy / min). After irradiation, incubate for 24h, then use CCK8 method to evaluate the activity. The irradiated cells were inoculated in 96-well plates at a density of 5x10 5 / mL (100 μL / 5 million / well), and CCK8 reagent was added to each well. After incubation in the dark for 3h, the absorbance at 450nm was measured using a microplate reader. Finally, data analysis was performed to determine the therapeutic effect of DMSS on neutrophils (statistical method: one-way ANOVA, compared with IR group, **P<0.01, ****P<0.0001).

[0052] The results are shown in Figure 2 B, after different concentrations of DMSS were used to treat HL-60 cells and irradiation with 5Gy dose, it was found that the number of HL-60 cells increased in a drug concentration dependent manner, indicating that the irradiation protection effect of DMSS was also reflected in the cell model.

[0053] 2.2 Cell apoptosis detection

[0054] The prepared drug DMSS was added to the cells for pre-treatment, and 1 h later, the cells were irradiated with 5 Gy X-rays (dose rate 2.046 Gy / min). After irradiation, the cells were incubated for 24 hours, and then the apoptosis was evaluated by Annexin V-FITC / 7-AAD double staining method. After irradiation, the cells were collected and stained with Annexin V-FITC and 7-AAD, and the apoptosis of the cells was analyzed by flow cytometry. The change of the apoptosis rate of the cells before and after drug treatment was analyzed, and the effect of DMSS on cell apoptosis was evaluated (statistical method: one-way ANOVA, compared with the IR group, **P<0.01, ***P<0.001, ****P<0.0001).

[0055] The results are shown in Figure 2 C and Figure 2 D, and it can be seen from the figure that the apoptosis rate of the cells in the non-drug irradiation group increased significantly, while the apoptosis rate of the cells in the drug irradiation group recovered significantly, indicating that DMSS can realize the irradiation protection effect on the cells by affecting the apoptosis of the cells.

[0056] 2.3 Detection of the expression level of apoptosis protein

[0057] In order to further verify the effect of DMSS on cell apoptosis, the present application uses Western blot (WB) technology to detect the expression changes of apoptosis-related proteins BCL-2 (anti-apoptosis protein) and BAX (pro-apoptosis protein).

[0058] First, the total protein of the cells treated by DMSS / irradiation was extracted; then, equal amounts of protein samples were separated by SDS-PAGE electrophoresis and transferred to a PVDF membrane; after the transfer was completed, 5% skim milk was used to block the membrane to block non-specific binding; then, specific anti-BCL-2 and BAX primary antibodies were added for incubation overnight, and the next day, TBST was used for washing, and HRP-labeled secondary antibodies were added for incubation; finally, ECL chemiluminescence reagent was used for color development, and the protein bands were detected by a gel imaging system, taking β-actin as an internal reference, and the relative expression levels of BCL-2 and BAX proteins were analyzed, so as to evaluate the regulation of DMSS on cell apoptosis (statistical method: one-way ANOVA, compared with the IR group, *P<0.05, **P<0.01, ***P<0.001).

[0059] The results are shown in Figure 2 E, Figure 2 F and Figure 2 G, and it can be seen from the figure that the expression of apoptosis-related protein BCL-2 decreases and the expression of BAX increases after irradiation, while the expression of BCL-2 increases and the expression of BAX decreases after drug administration, which further indicates that DMSS can realize the irradiation protection effect on the cells by affecting the apoptosis of the cells.

[0060] Example 3

[0061] This embodiment verifies the anti-radiation effect of DMSS through mouse experiments.

[0062] 3.1 Mouse grouping

[0063] Reference Figure 3 A, according to the principle of complete random design, mice were randomly divided into 3 test groups (survival test: n = 7; blood routine and flow analysis: n = 5): (1) blank control group (DMSO, no irradiation); (2) irradiation control group (DMSO + X-ray); (3) treatment group (DMSS 1 / 10 / 100 mg / kg + X-ray). Among them, DMSS is dissolved in 0.1% DMSO, and injected into mice intraperitoneally 6 hours before irradiation.

[0064] 3.2 Mouse survival analysis

[0065] After 1 hour of intraperitoneal administration, irradiation was performed with a lethal dose of 6 Gy X-rays (dose rate 2.046 Gy / min). After irradiation, the survival and body weight changes of each mouse were observed every day.

[0066] The survival situation is shown in Figure 3 B, as can be seen from the figure, the survival rate of mice in the administration group increased with the increase of the concentration of DMSS, while the mice in the non-administration group all died within 7 days (statistical method: one-way ANOVA, compared with the non-administration group, **P<0.01, ***P<0.001).

[0067] The body weight change results are shown in Figure 3 C, as can be seen from the figure, the body weight of mice in the non-administration group continuously decreased, while the body weight of mice in the administration group began to rise at 5-6 days (statistical method: one-way ANOVA, compared with the non-administration group, ***P<0.001). The results show that the irradiation protection effect of DMSS is also reflected in the mouse irradiation granulocyte reduction model.

[0068] 3.3 Mouse blood routine analysis

[0069] After 1 hour of intraperitoneal administration, irradiation was performed with a lethal dose of 6 Gy X-rays (dose rate 2.046 Gy / min). After irradiation, the survival and body weight changes of each mouse were observed every day.

[0070] Before blood collection, the mice were placed in the test environment for at least 30 minutes to adapt and reduce stress response; use a mouse fixer or manual fixation to ensure that the tail vein remains stable during the operation; use a 75% alcohol cotton ball to wipe the mouse tail, disinfect and wait for the alcohol to evaporate; immerse the mouse tail in warm water at about 42°C for 1-2 minutes to dilate the tail vein; cut the tail and let the blood flow into the blood collection tube.

[0071] After blood collection, the puncture site was gently pressed with a sterile cotton ball until hemostasis; the collected blood was added to a test tube containing an anticoagulant (such as EDTA), and gently mixed; finally, the peripheral blood neutrophil level was determined using a blood routine instrument (BC-5000 Vet, Mindray Animal Health Company, Shenzhen, China) (statistical method: one-way ANOVA, compared with the non-drug group, **P<0.01, ****P<0.0001).

[0072] The test results are shown in FIG. D, from which it can be seen that, compared with the blank group, the number of neutrophils in the non-drug irradiation group decreased significantly after sublethal dose irradiation, while the number of neutrophils in the drug irradiation group increased significantly with the increase of DMSS concentration; further verifying the irradiation protection effect of DMSS on neutrophils in the mouse irradiation model. Figure 3

[0073] 3.4 Flow cytometric analysis of hematopoietic cells in each line of mice

[0074] 3.4.1 Preparation of samples and single-cell suspensions

[0075] The mouse administration and irradiation conditions were the same as those in 3.3 blood routine analysis, and 15 days after irradiation, the mice were sacrificed by cervical dislocation and soaked in 75% ethanol; the skin was cut open, and the bilateral femur and tibia were aseptically removed, the muscle was removed, the epiphysis was cut off, and the bone marrow was washed with 1 mL needle head with IMDM medium (5 mL) from both ends of the bone shaft, repeated 2-3 times until the bone was white; filtered through a 70 μm filter, and the single-cell suspension was collected; at the same time, the complete spleen and thymus were taken out through the chest, and placed in a 6 cm dish containing 2 mL cold PBS, then a small piece of tissue was taken from each mouse, and the blunt end of the 1 mL syringe piston was gently ground, filtered through a 70 μm filter, and the single-cell suspension was collected.

[0076] Red blood cell lysate was directly added to the bone marrow and spleen single-cell suspension to 3 times the original volume, and the red blood cells were lysed on ice for 10 min, then centrifuged at 1000 rpm for 5 min, and the supernatant was discarded; resuspended in 5 mL FACS buffer and mixed well.

[0077] Each single-cell suspension was counted using a cell counter, and the results were recorded to calculate the required suspension volume for each staining tube. According to the calculated volume, each sample was separately dispensed into each staining tube, centrifuged at 5000 rpm for 1 min, the supernatant was discarded, and placed on ice.

[0078] 3.4.2 Antibody staining

[0079] ​Prepare the antibody mixture required for each cell staining scheme, MAC-1-APC labels myeloid cells, Gr1-PE labels neutrophils, and add the prepared antibodies to each staining tube respectively; the staining volume is 100 ul, and after 30 min on ice, add 100 ul FACS buffer to terminate the reaction, and then temporarily place in a 4 degree refrigerator.

[0080] 3.4.3 Flow detection and analysis

[0081] Evaluate the proportion and quantity changes of each line of hematopoietic cells in mice by flow cytometry (statistical method: single factor variance analysis, compared with the non-administration group, **P<0.01), and the results are shown in Figure 3 E and Figure 3 F, and it can be seen from the figure that the number of spleen myeloid cells in the non-administration irradiation group is significantly decreased compared with the blank group, while the number of spleen myeloid cells in the administration irradiation group is significantly increased with the increase of the concentration of DMSS, and reaches a peak at 10 mg / kg, indicating that DMSS has a radiation protection effect on myeloid hematopoietic cells in the mouse model, so it is feasible to extend the drug use of DMSS to the application of preventing HARS caused by radiation.

[0082] Figure 4 The statistical graphs of the effects of DMSS in zebrafish models, cell models and mouse models are shown in the figure, and it can be seen that DMSS has the function of reducing the apoptosis of neutrophils caused by radiation in the three models, so as to achieve the radiation protection effect.

[0083] In summary, the present application establishes a zebrafish model of neutropenia caused by radiation, and proves that the animal model can be used for large-scale primary screening of anti-radiation drugs, and the activity of the compound is verified by further cell and mouse tests, thereby forming a complete radiation protection drug screening and evaluation path, which significantly improves the research and development efficiency and the reliability of the results; secondly, DMSS belongs to flavonoids of natural origin, has excellent biocompatibility and safety basis, and such components exist widely in edible plants such as fruits and vegetables, and their antioxidant, anti-inflammatory and antitumor activities have been confirmed in various disease models, so DMSS may have lower toxic side effects and better clinical transformation potential compared with synthetic radiation protection agents; finally, the present application first discloses that DMSS realizes the protection effect by reducing the apoptosis of neutrophils caused by radiation in hematopoietic damage caused by radiation; at present, the types of radiation protection drugs are limited, and many of them have obvious side effects and other problems, DMSS as a new type of protection agent, its unique mechanism of action provides a new candidate molecule for the treatment of acute radiation damage of the hematopoietic system.

Claims

1. Application of the flavonoid 5-demethyl sweet orange flavonoid in the preparation of drugs for treating hematopoietic acute radiation syndrome.

2. The application according to claim 1, characterized in that: The 5-demethyl sweet orange flavonoids exert their radiation protection effect by influencing cell apoptosis.

3. The application according to claim 2, characterized in that: The 5-demethyl sweet orange flavonoids exert a radiation protection effect by reducing irradiation-induced neutrophil apoptosis.

4. The application according to claim 3, characterized in that: In the zebrafish irradiation neutropenia model, the 5-demethyl sweet orange flavonoids exerted an irradiation protection effect by alleviating the reduction in neutrophil count caused by irradiation, and the ability of the 5-demethyl sweet orange flavonoids to restore neutrophil count was concentration-dependent.

5. The application according to claim 3, characterized in that: In a cell irradiation model, the 5-demethyl sweet orange flavonoids exerted a protective effect on neutrophils by increasing the expression of the anti-apoptotic protein BCL-2 and decreasing the expression of the pro-apoptotic protein BAX.

6. The application according to claim 3, characterized in that: In a mouse irradiation model, the 5-demethyl sweet orange flavonoid has at least one of the following effects: (1) To alleviate or relieve the clinical symptoms of hematopoietic acute radiation syndrome; (2) Improve survival rate; (3) Reduce the number of apoptotic neutrophils in peripheral blood induced by radiation; (4) Reduce the number of irradiated myeloid cells in the spleen to induce apoptosis.

7. The application according to claim 1, characterized in that: The drug is a drug to be used before irradiation exposure.

8. The application according to claim 7, characterized in that: The dosage form of the drug is hard capsule, soft capsule, powder, granule, tablet, pill, honey ointment, oral liquid, suppository, tincture or injection.

9. The application according to claim 1, characterized in that: The drug also includes a pharmaceutically acceptable carrier.

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