Pharmaceutical composition for preventing and treating acute radiation sickness and application thereof

Abstract

The invention discloses a pharmaceutical composition for preventing and treating acute radiation sickness and application thereof. Animal experiments prove that the combined use of the estrogen anti-release drug and the thrombopoiesis promoting drug can obviously improve the survival rate of animals irradiated by lethal and even super lethal dose, can obviously increase the proportion and absolute quantity of hematopoietic stem cells and progenitor cells in bone marrow of the irradiated animals, and enhances the extramedullary hematopoietic activity of the spleen at the same time; and the recovery of hematopoietic functions of leukocytes, platelets and erythrocytes is promoted. According to the invention, the treatment effect on acute radiation sickness is remarkably improved by combining the estrogen drugs and the thrombopoiesis promoting drugs, and a new treatment scheme is provided for prevention and treatment of acute radiation sickness.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology. Specifically, this invention relates to a pharmaceutical composition for the prevention and treatment of acute radiation sickness and its application. Background Technology

[0002] With the rapid development of nuclear energy and technology, they have been widely applied in various fields such as industrial and agricultural production, medical and health care, scientific research, and national defense. This has led to an increasing exposure to ionizing radiation. While bringing immense convenience and benefits to humanity, these technologies also pose significant radiation hazards. During radiotherapy for cancer patients, while radiation kills tumor cells, it also causes severe damage to normal cells, often limiting the practical application of radiotherapy. Therefore, to combat radiation damage and ensure the safe and efficient application of nuclear energy and technology, it is necessary to vigorously develop anti-radiation damage medical technologies.

[0003] Estrogen drugs are a class of compounds that promote and maintain the development and maturation of female sex organs and secondary sexual characteristics. They are divided into two categories: natural and synthetic. Natural estrogen drugs include estradiol, estriol, and estrone, while synthetic estrogen drugs include ethinylestradiol, ethinylestradiol ether, norethinylestradiol, ethinylestradiol methyl ether, estradiol benzoate, and estradiol valerate. Thrombopoietin-stimulating drugs include recombinant human thrombopoietin (rhTPO) and thrombopoietin receptor agonists (TPO-RA). Currently, five thrombopoietin-stimulating drugs have been approved for clinical use worldwide, including rhTPO, romiplostim, eltrombopag, hetrombopag, avatrombopag, and lusutrombopag.

[0004] In the clinical treatment of patients with radiation accidents, multi-drug combination therapy can effectively promote functional recovery after severe bone marrow suppression, but the effects of combining estrogen drugs and thrombopoietin-stimulating drugs on radiation damage have not yet been disclosed. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the purpose of this invention is to provide a combination of drugs for the prevention and treatment of acute radiation sickness, which includes estrogen drugs and platelet-promoting drugs.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: The first aspect of the present invention provides a pharmaceutical composition for preventing and treating acute radiation sickness.

[0007] Furthermore, the pharmaceutical composition includes an estrogen and a thrombopoietin.

[0008] Furthermore, the estrogen drugs include any one or a combination of at least two of the following: estriol, solenoid estradiol, ethinylestradiol, ethinylestradiol, ethinylestradiol ether, ethinylestradiol methyl ether, estradiol, estradiol benzoate, "523" tablets and / or "500" injections.

[0009] Furthermore, the platelet-promoting drugs include any one or a combination of at least two of roprostine, eltrombopag, hetrombopag, avatrombopag, rutrombopag, TPO, and rhTPO.

[0010] Furthermore, the estrogen-like drug is selected from estriol and / or norestradiol.

[0011] Furthermore, the platelet-promoting drug is selected from roprostine and / or rhTPO.

[0012] Drug combination therapy can produce multi-level interactions, ultimately manifesting as synergistic, additive, or antagonistic effects. Synergistic effects refer to the phenomenon where the combined action of two or more drugs results in a consistent direction and enhanced efficacy, categorized as additive or enhancing effects. Antagonistic effects, on the other hand, refer to a weakening or disappearance of the combined effect. Additive or synergistic effects mean that the combined effect of two drugs acting on disease-related targets is equal to or greater than the sum of their individual effects.

[0013] In this invention, it was discovered for the first time that the combined use of estrogen-like drugs and thrombopoietin-promoting drugs can significantly improve the survival rate of lethal and even super-lethal irradiated mice, showing a synergistic effect. Moreover, the greater the irradiation dose, the more prominent the synergistic effect.

[0014] In some embodiments, the pharmaceutical composition further includes an estrogen or a pharmaceutically acceptable salt thereof and a thrombopoietin or a pharmaceutically acceptable salt thereof. The term "pharmaceutical acceptable salt" refers to salts of compounds of interest that are safe and effective for topical application in mammals and possess the desired biological activity. Pharmaceutically acceptable salts include salts containing acidic or basic groups present in the specified compounds. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochlorides, hydrobromides, hydroiodates, nitrates, sulfates, hydrogen sulfates, phosphates, acid phosphates, isonicotinate, acetates, lactates, salicylates, citrates, tartrates, pantothenates, hydrogen tartrate, ascorbic acid salts, succinates, maleates, gentisinates, fumarates, glucurons, glucuronides, sucrose salts, formates, benzoates, glutamates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, and papoate (i.e., 1,1'-methylene-di-(2-hydroxy-3-naphthylcarbamate)). Certain compounds used in this invention can form pharmaceutically acceptable salts with various amino acids. Suitable base salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. For a review of medicinal salts, see the following literature: BERGE et al., 66 J. PHARM. SCI. 1-19 (1977).

[0015] Furthermore, the pharmaceutical composition also comprises a pharmaceutically acceptable carrier and / or excipients.

[0016] Furthermore, the pharmaceutically acceptable carrier and / or excipients include any one or a combination of at least two of the following: diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

[0017] As used herein, the term "carrier and / or excipient" refers to a pharmaceutically acceptable material, composition, or vehicle, such as liquid or solid fillers, diluents, excipients, manufacturing excipients (e.g., lubricants, magnesium talc, calcium or zinc stearates, or stearic acid), or solvent encapsulation materials, that participates in the transport or translocation of a target compound from one part of an organ or organism to another part of an organism. Each carrier must be "acceptable" in the sense of compatibility with other components in the formulation and harmlessness to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, methyl cellulose, ethyl cellulose, microcrystalline cellulose, and cellulose acetate ester; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricants, such as magnesium stearate, sodium dodecyl sulfate, and talc; and (8) excipients, such as cocoa butter and suppository waxes. (9) Oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) Diols, such as propylene glycol; (11) Polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol (PEG); (12) Esters, such as ethyl oleate and ethyl laurate; (13) Agar; (14) Buffers, such as magnesium hydroxide and aluminum hydroxide; (15) Alginate; (16) Pyrogen-free water; (17) Isotonic saline; (18) Ringer's solution; (19) Ethanol; (20) pH buffer solution; (21) Polyesters, polycarbonates, and / or polyanhydrides; (22) Bulk fillers. Agents, such as polypeptides and amino acids; (23) serum components, such as serum albumin, HDL and LDL; (24) C2-C12 alcohols, such as ethanol; and (25) other non-toxic and compatible substances used in pharmaceutical preparations. Wetting agents, coloring agents, releasing agents, coating agents, sweeteners, flavoring agents, aroma agents, preservatives and antioxidants may also be present in the preparations.

[0018] Furthermore, the dosage ratio of the estrogen-like drug to the thrombopoietin-stimulating drug is 50:1.

[0019] In some embodiments, the dose ratio of the estrogen-like drug to the thrombopoietin-stimulating drug ranges from 0.1 to 500:1; in a specific embodiment of the present invention, the dose ratio of the estrogen-like drug to the thrombopoietin-stimulating drug is 50:1.

[0020] In some embodiments, the human dosage of the estrogen-like drug is 0.01 mg / kg to 1 mg / kg; the human dosage of the thrombopoietin-stimulating drug is 1 µg / kg to 2000 µg / kg; in a specific embodiment of the present invention, the human dosage of the estrogen-like drug is 0.5 mg / kg, and the human dosage of the thrombopoietin-stimulating drug is 10 µg / kg.

[0021] In some embodiments, the appropriate dosage of the pharmaceutical composition of the present invention can be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, condition, diet, administration time, route of administration, excretion rate and responsiveness. Skilled physicians can usually easily determine the prescription and the desired dosage that is effective for treatment.

[0022] Furthermore, the estrogen-like drugs and thrombopoietin-stimulating drugs are administered simultaneously or sequentially.

[0023] In some embodiments, the estrogen and thrombopoietin in the pharmaceutical composition of the present invention can be administered simultaneously or sequentially. When administered sequentially, the administration method includes: administering the estrogen first, followed by the thrombopoietin; or administering the thrombopoietin first, followed by the estrogen. In a specific embodiment of the present invention, the estrogen in the pharmaceutical composition is administered before irradiation, and the thrombopoietin is administered after irradiation.

[0024] Furthermore, the dosage forms of the pharmaceutical composition include non-gastrointestinal dosage forms and gastrointestinal dosage forms.

[0025] Furthermore, the non-gastrointestinal dosage forms include injectable dosage forms, respiratory dosage forms, cavity dosage forms, mucosal dosage forms, and skin dosage forms.

[0026] Furthermore, the gastrointestinal dosage forms include tablets, granules, capsules, solutions, dry suspensions, powders, sustained-release preparations, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0027] In some embodiments, the injectable dosage forms include, but are not limited to, various injectables such as intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, and intracavitary injections; the respiratory dosage forms include, but are not limited to, sprays, aerosols, and powder inhalers; the cavity dosage forms include, but are not limited to, suppositories, aerosols, effervescent tablets, drops, and pills, for use in the rectum, vagina, urethra, nasal cavity, and ear canal; the mucosal dosage forms include, but are not limited to, eye drops, nasal drops, ointments, mouthwashes, sublingual tablets, adhesive tablets, and patches; and the skin dosage forms include, but are not limited to, topical solutions, lotions, liniments, ointments, plasters, pastes, and patches.

[0028] A second aspect of the present invention provides a medicine box.

[0029] Furthermore, the medicine box includes: (I) The first preparation containing estrogen-like drugs; (II) A second formulation containing a platelet-producing drug.

[0030] Furthermore, the first formulation is selected from estriol, solenoid estradiol, ethinylestradiol, ethinylestradiol, ethinylestradiol ether, ethinylestradiol methyl ether, estradiol, estradiol benzoate and / or estradiol valerate, "523" tablets and / or "500" injections. Furthermore, the first formulation is selected from estriol and / or norestradiol.

[0031] Furthermore, the second formulation is selected from roprostine, eltrombopag, haitrombopag, avatrombopag, rutrombopag, TPO and / or rhTPO; Furthermore, the first formulation is selected from roprolastine and / or rhTPO; Preferably, the medicine box also includes an instruction manual; Preferably, the instruction manual specifies that the first and second preparations are used in combination to prevent and / or treat acute radiation sickness.

[0032] In this invention, the terms "first active ingredient," "estrogenous drug," or "first formulation" are used interchangeably and refer to an estrogenous drug. The active ingredient of this invention can be any crystalline form, amorphous form, dehydrated form, solvate, hydrate, or enantiomer of a pharmaceutically acceptable estrogenous drug. In this invention, the estrogenous drug refers to the first formulation of this invention.

[0033] In this invention, the terms "second active ingredient," "second formulation," or "thrombopoietin drug" are used interchangeably, referring to thrombopoietin drugs, specifically including roprostine, eltrombopag, hetrombopag, avatrombopag, rutrombopag, TPO, and rhTPO. The active ingredient of this invention can be any crystalline form, amorphous form, dehydrated form, solvate, hydrate, or enantiomer of a pharmaceutically acceptable thrombopoietin drug. In this invention, the thrombopoietin drug refers specifically to the second formulation of this invention.

[0034] When used in a kit, the term "instructions for use" as used herein includes: publications, records, icons, or any other medium of expression that may be used to convey the effectiveness of the kit for its intended purpose. For example, instructions for use may be affixed to or included in the container of the kit.

[0035] The third aspect of the present invention provides any of the following applications: (1) The application of combined estrogen drugs and thrombopoietin drugs in the preparation of drugs for treating acute radiation sickness; (2) Application of estrogen drugs in the preparation of drugs that enhance the therapeutic effect of thrombopoietin-stimulating drugs on acute radiation sickness; (3) Application of platelet-producing drugs in the preparation of drugs that enhance the therapeutic effect of estrogen drugs on acute radiation sickness.

[0036] (4) The use of combined estrogen drugs and thrombopoietin drugs in the preparation of drugs for the prevention and / or treatment of bone marrow suppression caused by radiation exposure; (5) The use of a combination of estrogen-like drugs and thrombopoietin-promoting drugs in the preparation of drugs for the prevention and / or treatment of death in animals or humans caused by radiation exposure; (6) The use of estrogen drugs and thrombopoietin drugs in combination in the preparation of drugs for the prevention and / or treatment of peripheral blood pancytopenia, leukopenia, neutropenia, lymphopenia, thrombocytopenia and / or anemia caused by radiation exposure; (7) Application of combined estrogen drugs and platelet-producing drugs in the preparation of drugs for the prevention and / or treatment of damage to hematopoietic stem and / or progenitor cells caused by radiation.

[0037] Furthermore, the estrogen drugs include any one or a combination of at least two of the following: estriol, norethinylestradiol, ethinylestradiol, ethinylestradiol ether, ethinylestradiol methyl ether, estradiol, estradiol benzoate and / or estradiol valerate, "523" tablets and / or "500" injections.

[0038] Furthermore, the platelet-promoting drugs include any one or a combination of at least two of roprostine, eltrombopag, hetrombopag, avatrombopag, rutrombopag, TPO, and rhTPO.

[0039] Furthermore, the estrogen-like drug is selected from estriol and / or norestradiol.

[0040] Furthermore, the platelet-promoting drug is selected from roprostine and / or rhTPO.

[0041] Furthermore, the acute radiation sickness includes bone marrow type acute radiation sickness, intestinal type acute radiation sickness, and cerebral type acute radiation sickness.

[0042] In this invention, the treatment refers to the act of preventing and reducing the occurrence or development of a disease, thereby inhibiting, suppressing, alleviating, improving, slowing down, stopping, delaying, or reversing the progression or aggravation of the disease. The various indicators of maintaining and / or treating the disease, disorder, or pathological state at the time of medication include alleviating or reducing the symptoms or complications of a specific disease, or curing or eliminating the disease, disorder, or condition. Therefore, the treatment of acute radiation sickness described in this invention includes the prevention, relief, and / or treatment of acute radiation sickness.

[0043] Furthermore, the present invention also provides a method for preventing and treating acute radiation sickness, the method comprising the steps of administering an effective amount of the pharmaceutical composition of the first aspect of the present invention to a subject in need.

[0044] In this invention, the effective amount refers to the amount of compound that effectively produces the desired preventive, alleviating, or therapeutic effect. The amount of the pharmaceutical composition of this invention required to achieve an effective amount may vary depending on factors such as the compound, the symptoms and their severity, and the age of the mammal being treated. However, the specific dosage can be conventionally determined by those skilled in the art based on their knowledge in the field and in conjunction with the disclosure of this invention.

[0045] In some embodiments, the drug composition can be administered to a patient via oral administration, injection, or topical administration. For example, the method may include administering the drug composition to the subject three times a day, once a day, or every two days. In some embodiments, injection administration may include subcutaneous injection, intramuscular injection, intravenous injection, etc. In some embodiments, injection administration may include injecting the drug composition directly into or near a lesion. In some embodiments, topical administration may include rectal administration, nasal administration, ear administration, intramedullary administration, intra-articular administration, intrapleural administration, etc., or any combination thereof. In some embodiments, the drug composition may be administered to the subject via a combination of different routes of administration.

[0046] In some embodiments, the subject of the pharmaceutical composition is an animal, preferably a mammal (human and non-human animals), including but not limited to: humans, non-human primates (especially higher primates, such as macaques, cynomolgus monkeys, stump-tailed macaques, rhesus monkeys, shrews, golden snub-nosed monkeys, and tree shrews), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, cattle, any livestock or pets, etc. In a preferred embodiment, the subject is preferably a human.

[0047] Advantages and beneficial effects of the present invention: This invention is the first to discover that the combination of estrogen-based drugs and thrombopoietin-stimulating drugs has a synergistic effect against acute radiation sickness. Animal experiments demonstrate that the combined use of estrogen-based anti-radiation drugs such as norethisterone and estriol with thrombopoietin-stimulating drugs such as roprostine and rhTPO significantly improves the survival rate of animals exposed to lethal and even super-lethal doses of radiation. It also significantly increases the proportion and absolute number of hematopoietic stem and progenitor cells in the bone marrow of irradiated animals, enhances extramedullary hematopoietic activity in the spleen, and promotes the recovery of hematopoietic function of the leukocyte, platelet, and erythrocyte lineages. This invention, by combining estrogen-based drugs and thrombopoietin-stimulating drugs, significantly improves the therapeutic effect on acute radiation sickness, providing a new treatment option for the prevention and treatment of acute radiation sickness. Attached Figure Description

[0048] Figure 1 The effect of combined administration of estriol and roprolastine on the survival and body weight of mice irradiated with different doses was investigated; among which, Figure 1 A shows the survival and weight of mice irradiated with 10.0 Gy; Figure 1 B shows the survival and weight of mice irradiated with 10.5 Gy; Figure 1 C shows the survival and weight of mice irradiated with 11.0 Gy; Figure 2 The effects of combined administration of estriol and roprolastine on survival and peripheral blood cell recovery in mice subjected to 9.0 Gy lethal irradiation were investigated; among which, Figure 2 A shows the 30-day survival results; Figure 2 BG shows the results of white blood cell, lymphocyte, neutrophil, red blood cell, hemoglobin and platelet levels, respectively; * indicates comparison with the irradiation group, # indicates comparison with the combined drug administration group; Figure 3 The effect of combined administration of estriol and roprolastine on the recovery of peripheral blood counts in mice subjected to 6.5 Gy sublethal irradiation; among which, Figure 3 AF shows the results of white blood cell, neutrophil, platelet, lymphocyte, red blood cell and hemoglobin levels, respectively; * indicates comparison with the irradiation group, # indicates comparison with the combined drug administration group; Figure 4The effect of combined administration of estriol and roprostine on bone marrow hematopoietic recovery in mice 10 days after 6.5 Gy irradiation; among which, Figure 4 The images show the bone marrow H&E staining results of normal unirradiated mice, irradiated mice, mice irradiated with estriol alone, mice irradiated with roprostine alone, and mice irradiated with a combination of roprostine and estriol. Figure 4 F shows the bone marrow nucleated cell count; * indicates comparison with the irradiation group, # indicates comparison with the combined drug administration group; Figure 5 The effect of combined administration of estriol and roprostine on the recovery of bone marrow hematopoietic stem and progenitor cells 10 days after 6.5 Gy irradiation in mice; among which, Figure 5 A displays the door-drawing strategy; Figure 5 BH shows the absolute values ​​of LK, LSK, MMP4, MMP3, MMP2, ST-HSC, and LT-HSC cell populations and their proportions of the total number of nucleated cells in the bone marrow, respectively. Figure 5 I shows the proportion of each cell population in the LSK partition; * indicates comparison with the irradiated group, # indicates comparison with the combined drug administration group; Figure 6 The effect of combined administration of estriol and roprostine on extramedullary hematopoiesis in the spleen of mice 10 days after 6.5 Gy irradiation; among which, Figure 6 A shows the size of the spleen; Figure 6 B shows the spleen coefficient; Figure 6 C shows the number of nucleated cells in the spleen; Figure 6 D shows spleen sections stained with HE; Figure 7 The effect of combined administration of estriol and roprostine on the recovery of spleen hematopoietic stem and progenitor cells 10 days after 6.5 Gy irradiation in mice; among which, Figure 7 A displays the door-drawing strategy; Figure 7 BH displays the absolute values ​​of LK, LSK, MMP4, MMP3, MMP2, ST-HSC, and LT-HSC cell populations and their proportions of nucleated cells in the spleen, respectively; * indicates comparison with the irradiation group, and # indicates comparison with the combined drug administration group; Figure 8 The effects of combined administration of estriol and rhTPO on the survival and body weight of mice subjected to 10.0 Gy superlethal irradiation were investigated. Figure 8 A shows the survival of mice 30 days after irradiation; Figure 8 B shows the change in mouse body weight 30 days after irradiation; Figure 9 The effects of combined administration of estriol and rhTPO on survival and peripheral blood cell recovery in mice subjected to 9.0 Gy lethal irradiation were investigated; among which, Figure 9 A indicates a 30-day survival rate; Figure 9BF shows changes in white blood cells, neutrophils, platelets, red blood cells, and hemoglobin; * indicates comparison with the irradiation group, # indicates comparison with the combined drug administration group; Figure 10 The effects of combined administration of nystatin and roprostine on the survival and body weight of mice subjected to 10.0 Gy superlethal irradiation were investigated. Figure 10 A shows the changes in mouse survival 30 days after irradiation; Figure 10 B shows the change in mouse body weight 30 days after irradiation. Detailed Implementation

[0049] The present invention will now be described in detail with reference to specific embodiments and accompanying drawings. In the following description, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments can be modified in various ways without departing from the spirit or scope of the invention. Therefore, the drawings and description are considered exemplary in nature and not restrictive. Unless otherwise specified, specific conditions in the following embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments used, unless otherwise specified, are all commercially available products.

[0050] The experimental materials and methods used in specific embodiments of the present invention are as follows: 1. Laboratory animals Male C57BL / 6J mice aged 6-8 weeks were purchased from Beijing Jicui Yaokang Biotechnology Co., Ltd. The purchased mice were randomly assigned to cages, with no more than 5 mice per cage, and were housed in an SPF (specific pathogen-free) barrier environment in the animal laboratory of the Experimental Animal Center of the Academy of Military Medical Sciences. During the experiment, the animals had free access to rat and mouse maintenance pellet feed (…). 60 Sterilized (CO2) water and acidified water (pH 2.5-3.0) were used. Pelleted feed was purchased from Beijing Huafukang Biotechnology Co., Ltd. The rearing environment was maintained with a 12-hour light / 12-hour dark cycle, temperature 23±2°C, and relative humidity 50±20%. Mice were acclimatized to the environment for at least 7 days before the experiment. All animal experiments were approved by the Laboratory Animal Welfare and Ethics Committee of the Academy of Military Medical Sciences and strictly followed the institution's laboratory animal operation guidelines. The "Beijing Municipal Regulations on the Management of Laboratory Animals" were strictly followed, and mice were euthanized according to the "Guidelines for Laboratory Animal Welfare and Ethics Review" designated by the General Administration of Quality Supervision, Inspection and Quarantine of China. The mice underwent cervical dislocation after CO2 inhalation.

[0051] 2. The experimental reagents and antibody information used in the following examples are shown in Table 1.

[0052] Table 1. Information on experimental reagents and antibodies

[0053] 3. Experimental Methods (1) Drug preparation and administration Preparation and subcutaneous administration of rhTPO: rhTPO is provided in liquid form at a concentration of 100 µg / mL and stored at 4°C protected from light. See the examples for drug preparation concentrations.

[0054] Preparation and subcutaneous administration of roprostine: Roprostine is provided in solid powder form. Before use, add 0.72 mL of physiological saline to each vial of solid powder, gently shake to completely dissolve into a colorless liquid with a concentration of 500 μg / mL, aspirate with a syringe and place in a 1.5 mL centrifuge tube, aliquot, and store at -80℃ protected from light. When using, thaw and centrifuge before use, avoiding repeated freeze-thaw cycles. Gently shake to mix, and administer subcutaneously at a concentration of 100 ug / kg. The solvent is sterile physiological saline, and the dosage is 0.2 mL per mouse.

[0055] Estriol: White solid, store at -20℃. Dosage: 5 mg / kg, in a solvent of 1 / 33 benzyl alcohol and 32 / 33 tea oil (dissolve completely in benzyl alcohol first, then in tea oil), 0.2 ml intraperitoneally or 0.1 ml intramuscularly. The solvent control group received the same volume of solvent. Nereestrol: White solid, store at 4°C. Dosage: 5 mg / kg, dissolved in polyethylene glycol 400, administered by gavage, 0.2 ml per mouse. The solvent control group received an equal volume of solvent. (2) Animal irradiation All irradiation experiments were conducted between 9:00 AM and 11:00 AM. Before irradiation, unanesthetized mice were placed in a well-ventilated restraint device consisting of 20 independent compartments separated by plexiglass partitions. A cobalt-60 gamma-ray source from the Irradiation Center of the Institute of Radiation Medicine, Academy of Military Medical Sciences was used. 60 Whole-body irradiation with Co gamma rays was performed. The cobalt source dose rate was measured using a PTW UNIDOS dosimeter connected to a finger-shaped ionization chamber (300130.6cc). The irradiation time was determined based on the decay of the cobalt source over time and the attenuation factors of air and tissue during irradiation. The whole-body irradiation doses in this study were 6.5 Gy, 9 Gy, 10 Gy, 10.5 Gy, and 11 Gy, with a dose rate of approximately 40 Gy / min.

[0056] (3) Study on the survival of irradiated animals Before the experiment, mice whose body weight deviated from the population mean by 10% were excluded, and they were randomly assigned to groups. Four to five experimental groups were set up for each dose, with 10 mice in each group, identified by the number of their tail tattoos. See the example for grouping details. 60Mice were observed daily for 30 days following Co-γ irradiation. Mice were weighed on the day of irradiation and on days 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 24, 27, and 30 post-irradiation. Survival data were plotted as Kaplan-Meier curves using GraphPad Prism 10.1.2 software. Mice were euthanized at the end of the experiment following the above procedure.

[0057] (4) Evaluation of drug synergistic effects This patent uses the King's equation to calculate the Q value, which evaluates the synergistic effect of combined drug use. The equation is: Q = E (A + B) / (E A + E B - E A × E B ), where E A and E B E represents the survival rate of a single drug. (A + B) The Q value represents the survival rate of combined drug therapy. The criteria for Q value determination are as follows: when Q < 0.85, it indicates antagonistic effect; when 0.85 ≤ Q < 1.15, it indicates additive effect; when Q ≥ 1.15, it indicates synergistic effect.

[0058] (5) Hematological studies of irradiated animals Mice irradiated with 6.5 Gy were given blood samples (10 μL each) from their tail veins at 7 and 10 days post-irradiation, and mice irradiated with 9 Gy were given blood samples at 7, 10, 14, and 18 days post-irradiation. After blood collection, mice were returned to their original cages for further observation, provided no abnormal bleeding was observed at the collection site. The blood samples were mixed with 2 mL of hematology analyzer diluent (MEK-640) and then used in pre-dilution mode on a Celltac ES automated hematology analyzer (NIHON KOHDEN) for complete blood cell count and white blood cell differential, counting red blood cells, platelets, white blood cells, lymphocytes, monocytes, and neutrophils.

[0059] (6) Flow cytometry analysis of bone marrow and spleen hematopoietic stem and progenitor cells (HSPCs) Mice irradiated with 6.5 Gy were sacrificed on day 10 post-irradiation. The mice were dissected, and the femur and spleen were separated. Bone marrow and spleen were extracted. Hematopoietic cells from the femur were flushed out using RPMI-1640. The spleen was ground to prepare a single-cell suspension. The prepared single cells from the bone marrow and spleen (6 × 10⁶ cells) were then... 6Cells were first treated with an anti-mouse Fc receptor blocker (TruStainFcX™ (anti-mouse CD16 / 32)) to block the non-specific binding of antibody Fc fragments to cells. Antibodies including Lineage cocktailPerCP-Cy5.5, Sca-1 PE-Cy7, C-kit APC-H7, CD150 PE, CD48 FITC, and CD135 APC were added at a 1:100 ratio to DPBS (2% FBS) to prepare an antibody mixture. The antibody sources are as described in the table above. Cells were incubated with the antibody mixture at 4°C for 30 minutes, followed by the addition of cell lysis buffer. After 5 minutes, the cells were washed with DPBS (2% FBS), and finally, a certain amount of DPBS (2% FBS) was added. The mixture was then filtered through a 40 μm sieve into flow cytometry tubes containing 7-AAD. Data were then analyzed using a BD FACS Verse™ flow cytometer and BD FACSuite software. The cell populations analyzed included: lymphoid progenitor cells (LK: Lineage-Sca-1-C-Kit+), hematopoietic stem / progenitor cells (LSK: Lineage-Sca-1+C-Kit+), pluripotent progenitor cells 4 (MPP4: Lineage-Sca-1+C-Kit+CD135+), pluripotent progenitor cells 3 (MPP3: Lineage-Sca-1-C-Kit+CD135-CD48+CD150-), pluripotent progenitor cells 2 (MPP2: Lineage-Sca-1-C-Kit+CD135-CD48+CD150+), long-term hematopoietic stem cells (LT-HSC: Lineage-Sca-1-C-Kit+CD135-CD48-CD150+), and short-term hematopoietic stem cells (ST-HSC: Lineage-Sca-1-C-Kit+CD135-CD48-CD150-).

[0060] (7) Statistical analysis All experimental data were processed using GraphPad Prism 10.0 software (GraphPad Software Inc., USA), and data are expressed as mean ± standard deviation (Mean ± SD). Differences between groups were analyzed using a two-tailed Student's t-test; survival analysis was performed using the Kaplan-Meier method to plot survival curves. Statistical significance was defined as follows: compared with the irradiation group: *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; compared with the combined treatment group: #P<0.05, ##P<0.01, ###P<0.001, and ####P<0.0001.

[0061] Example 1: Combined administration of estriol and roprostine synergistically protects mice from lethal doses of gamma radiation. To investigate whether pre-irradiation prophylaxis with estriol and post-irradiation treatment with a high dose of roprostine produce additive or synergistic protective effects, we first tested this in a mouse model subjected to a 10.0 Gy lethal dose total body irradiation (TBI). The experiment included a solvent control group, an estriol prophylaxis group, a roprostine treatment group, and an estriol + roprostine combination group. The solvent control group received the same experimental procedures and total solvent volume as the estriol + roprostine combination group, administering either estriol or roprostine solvent at the corresponding administration times. Estriol was administered intraperitoneally at 5 mg / kg 48 hours before irradiation; roprostine was administered subcutaneously at 100 µg / kg 30 minutes after irradiation. Figure 1 As shown in Figure A, all mice in the solvent control group died within 13 days after irradiation. The 30-day survival rates of mice in the roprostine group, estriol group, and estriol + roprostine group were 50%, 80%, and 100%, respectively. Based on the King's formula, we found that the Q value was 1.11 when estriol and roprostine were used in combination.

[0062] Subsequently, we further evaluated the synergistic effect of the combined use of estriol and roprostine in a mouse model irradiated with a superlethal dose of 10.5 Gy. Figure 1 As shown in Figure B, mice in both the solvent control group and the roprostine group experienced a sustained decrease in body weight after irradiation. All mice in the solvent control group died within 13 days of irradiation, while the 30-day survival rate in the roprostine group was only 10%. Mice in the estriol group experienced a sustained decrease in body weight starting 13 days after irradiation, reaching its lowest point at 19 days, and then began to recover; the 30-day survival rate was 80%. The estriol + roprostine group showed no significant decrease in body weight within 30 days of irradiation, and the 30-day survival rate was 100%. The Q value was 1.22, demonstrating a synergistic effect.

[0063] When the total body radiation dose increases to 11.0 Gy, such as Figure 1 As shown in Figure C, mice in the solvent control group, roprostine group, and estriol group all experienced a continuous decrease in body weight after irradiation and died within 14, 18, and 13 days after irradiation, respectively. In contrast, mice in the estriol + roprostine group began to lose body weight on day 7 after irradiation, reaching a stable weight by day 21, with a 30-day survival rate of 70%. The Q value was positive infinity, demonstrating a significant synergistic effect.

[0064] The above results indicate that combined administration of estriol and roprostine can significantly improve the survival rate of lethal and even superlethal irradiated mice, demonstrating a synergistic effect. Moreover, the higher the irradiation dose, the more prominent the synergistic effect.

[0065] Example 2: Combined administration of estriol and roprostine significantly promoted the recovery of multilineage hematopoiesis in irradiated mice. To evaluate the effect of combined administration of estriol and roprostine on hematological recovery in irradiated mice, we compared the effects of estriol and roprostine alone or in combination on peripheral blood count recovery on days 7, 10, 14, and 18 post-irradiation in a 9.0 Gy whole-body irradiation mouse model. Figure 2 As shown, the peripheral blood counts of mice in the solvent control group decreased significantly as expected after irradiation, reaching their lowest point at 10 or 14 days post-irradiation. Mortality began 10 days after irradiation, and all mice died within 17 days. In the estriol and roprostatin monotherapy and combination therapy groups, all peripheral blood count indicators reached their lowest points at 7 or 10 days post-irradiation, respectively, and then began to recover. The combination therapy group had essentially returned to pre-irradiation levels by 14 days post-irradiation, while the monotherapy groups had not yet returned to pre-irradiation levels by 18 days post-irradiation. In the estriol group, 2 out of 10 mice died, while all mice in the other two groups survived. On day 7 post-irradiation, only the combined drug group showed a significantly higher peripheral blood leukocyte count than the control group. On day 10 post-irradiation, some peripheral blood indicators in the roprostine group, such as leukocytes, neutrophils, erythrocytes, and hemoglobin, as well as all peripheral blood indicators in the combined drug group, were significantly higher than those in the solvent control group. By day 14 post-irradiation, all peripheral blood indicators in mice treated with estriol and roprostine alone and in combination, except for lymphocytes in the single drug group, were significantly higher than those in the solvent control group. Specifically, the leukocyte, erythrocyte, and platelet counts in the combined drug group were 3.3, 3.1, and 74.4 times, 2.4, 1.6, and 4.7 times, and 7.6, 5.9, and 36.0 times, respectively, compared to the estriol group, roprostine group, and solvent control group. Subsequently, we further evaluated the promoting effect of combined administration of estriol and roprostine on hematological recovery in irradiated mice using a 6.5 Gy sublethal dose irradiation mouse model. Figure 3 As shown, the number of white blood cells, lymphocytes, and platelets in the estriol + roprostine combination group on day 7 after irradiation and the number of platelets on day 10 after irradiation were significantly higher than those in the estriol and roprostine monotherapy group and the solvent control group (P<0.01).

[0066] In conclusion, the combined administration of estriol and roprostine can significantly promote the recovery of hematopoiesis in lethal and sublethal irradiated mice, and its efficacy is significantly better than that of estriol or roprostine alone. Moreover, the higher the irradiation dose, the more significant the therapeutic advantage of the combined administration.

[0067] Example 3: Combined administration of estriol and roprostine significantly promoted the regeneration of hematopoietic stem and progenitor cells in the bone marrow of irradiated mice. Total body irradiation (TBI)-induced bone marrow suppression is a pathological mechanism leading to pancytopenia, primarily manifested as a significant reduction in the number of bone marrow cells. Based on this, we compared the effects of estriol and roprostine, alone or in combination, on the recovery of bone marrow hematopoiesis on day 10 after 6.5 Gy total body irradiation in mice. Figure 4As shown, longitudinal sections of the femur from mice in the solvent control group irradiated with 6.5 Gy revealed a significant decrease in cell density, a reduction in the number of megakaryocytes, and the presence of adipocytes filling the bone marrow cavity, indicating severe impairment of bone marrow hematopoietic function. In contrast, femur sections from mice receiving estriol and roprostine, alone or in combination, showed more hematopoietic cell regeneration microfoci, increased cell density, and a greater number of megakaryocytes, indicating a significant recovery of bone marrow hematopoietic function. The estriol and roprostine combination group showed better bone marrow recovery on day 10 post-irradiation than the single-drug groups. Consistent with histological observations, the number of bone marrow nucleated cells (BMNCs) in the estriol, roprostine, and estriol + roprostine groups was significantly higher than that in the solvent control group, being 1.9 times, 2.0 times, and 2.4 times higher, respectively.

[0068] Hematopoietic stem cells (HSCs) and multipotent progenitor cells (MPPs) together constitute the origin of all types of blood cells and play a central role in maintaining hematopoietic homeostasis and regeneration. Ionizing radiation exposure can lead to a significant reduction in the number of hematopoietic stem / progenitor cells, thereby impairing the regenerative capacity of bone marrow. Based on the synergistic effect of estriol combined with roprostine in improving hematopoietic function and enhancing the survival rate of animals after radiation exposure, this study further explored the regulatory effect of this combined strategy on the regeneration of HSCs and MPPs in the bone marrow of mice after whole-body irradiation.

[0069] Following the methods described in previous literature, we used multiparameter flow cytometry to perform phenotypic analysis on mouse bone marrow cells that had undergone whole-body irradiation with 6.5 Gy and different treatments. In the mouse hematopoietic system, HSCs were mainly enriched in Lin. - Sca-1 + c-Kit + (LSK) cell population, collectively referred to as hematopoietic stem / progenitor cells (HSPCs). For example... Figure 5 As shown, the percentage and absolute number of LK and LSK cells in the bone marrow of mice in each treatment group were significantly higher than those in the solvent control group. The combination of estriol and roprostine showed the most significant effect, with the absolute numbers of LK and LSK cells in the bone marrow being 3.3, 1.9, and 69.8 times, and 2.0, 1.8, and 36.4 times, respectively, compared to the estriol group, roprostine group, and solvent control group.

[0070] Radiation can induce myeloid-biased pluripotent progenitor cells (MPPs) such as MPP3 (phenotype CD135) in the LSK compartment of the bone marrow. - CD48 + CD150 +LSK) and MPP2 (phenotype CD135) - CD48 + CD150 - The proportion of LSK was significantly increased, and lymphoid-biased MPP4 (phenotype CD135) was observed. + CD150 - The ratio of LSK (low-speed leukocyte-weighted stem cells) decreased sharply, and short-term hematopoietic stem cells (HSCST, phenotype CD135) decreased significantly. - CD48 - CD150 - The ratio of LSK (low-lying sclerosing leukocytes) increased significantly, and long-term hematopoietic stem cells (HSCLT, phenotype CD135) were also observed. - CD48 - CD150 + The LSK ratio was reduced. Pre-irradiation administration of estriol or post-irradiation administration of roprost significantly reversed the radiation-induced increase in the HSCST ratio and decrease in the MPP4 ratio, and increased the MPP2 ratio to varying degrees. Estriol had the strongest effect on increasing the MPP2 ratio, while roprost had the best effect on increasing the MPP4 ratio. The combination of estriol and roprost reduced the MPP2 ratio and increased the HSCLT ratio to varying degrees compared to administration alone.

[0071] Consistent with changes in the ratios of MPPs and HSCs within the LSK region of bone marrow, administration of estriol before irradiation or roprostine after irradiation significantly increased the proportion and absolute number of MPP3 and MPP2 nucleated cells in the bone marrow of irradiated mice, and significantly decreased the proportion and number of HSCSTs. Compared to the solvent control group, the proportion and number of MPP4 and HSCLTs were significantly increased in the bone marrow of the roprostine group but not the estriol group. It was found that the combined administration of estriol and roprostine significantly increased the number of HSCLTs, MPP4, and MPP3, as well as the proportion of HSCLTs and MPP4s, compared to administration of either alone. Specifically, the absolute number of MPP4s in the combined group was 4.7 times, 1.8 times, and positive infinity, respectively, compared to the estriol group, roprostine group, and solvent control group; the absolute number of HSCLTs was 9.3 times, 2.8 times, and 71.2 times, respectively, compared to the estriol group, roprostine group, and solvent control group.

[0072] In summary, both pre-treatment administration of estriol and post-treatment administration of roprostine significantly promoted the recovery of bone marrow hematopoietic stem and progenitor cells. Estriol was more favorable for myeloid cell recovery, while roprostine was more favorable for lymphoid cell recovery. It was found that the combined administration of estriol and roprostine, compared with their individual administration, significantly enhanced the long-term reconstitution of bone marrow hematopoietic stem cells and lymphoid cell differentiation, demonstrating a stronger hematopoietic recovery-promoting effect.

[0073] Example 4: Combined administration of estriol and roprostine significantly enhanced extramedullary hematopoietic function of the spleen in irradiated mice. When factors such as radiation suppress bone marrow hematopoiesis, hematopoietic stem cells (HSCs) in the bone marrow can be induced to migrate to the spleen, thereby initiating extramedullary hematopoiesis (EMH). Extramedullary hematopoiesis in the spleen plays a crucial role in the reconstitution of the hematopoietic system after radiation injury. For example... Figure 6 As shown, on day 10 after irradiation, the spleen volume and nucleated cell count of mice treated with 6.5 Gy TBI were significantly reduced. Both prophylactic administration of estriol and post-irradiation administration of roprostine significantly alleviated the radiation-induced decrease in spleen coefficient and nucleated cell count, with estriol showing a slightly better effect than roprostine. Notably, the combined administration of estriol and roprostine significantly improved the hematopoietic status of the spleen, resulting in a significant increase in spleen volume on day 10 after irradiation. The spleen coefficient was significantly higher than that of each single-drug group and the solvent control group, and the number of nucleated cells in the spleen was 1.5 times, 1.9 times, and 6.0 times that of the estriol group, roprostine group, and solvent control group, respectively.

[0074] Histopathological analysis showed that after irradiation, the white and red pulp of the spleen in mice was structurally disordered and poorly defined, splenic corpuscles were atrophied and sparsely arranged, the red pulp splenic sinuses were dilated and congested, and the number of hematopoietic cells was reduced. In mice treated with estriol and roprostine alone or in combination, the red pulp region of the spleen was significantly expanded due to infiltration of a large number of hematopoietic cells, and megakaryocytes were visible, indicating active extramedullary hematopoiesis in the spleen, with the combined administration group showing particularly significant effects.

[0075] To further evaluate the promoting effect of combined administration of estriol and roprostine on the proliferation of splenic hematopoietic stem / progenitor cells (HPSCs), we used flow cytometry to analyze the changes in various HPSC subsets in the mouse spleen on day 10 after irradiation. Figure 7 As shown, the proportion and number of LSK cells in the spleen of mice were significantly reduced after irradiation, and LK cells were almost undetectable. Administration of estriol and roprostine alone significantly increased the proportion and number of LK and LSK cells; compared with individual administration, combined administration significantly increased the proportion or number of LK cells in the spleen of mice, with the absolute number of LK cells being 4.3 times and 2.4 times that of the estriol group and the roprostine group, respectively, while having no significant effect on the proportion or number of LSK cells.

[0076] Compared to the solvent control group, estriol and roprostine, alone or in combination, significantly increased the MPP2 ratio and decreased the MPP3 ratio in the LSK compartment of the spleen, and increased the HSCLT ratio and decreased the HSCST ratio to varying degrees, but there were no statistically significant differences among the treatment groups. Consistent with these changes, the proportion and absolute number of MPP3 in nucleated cells of the spleen of mice in each treatment group, as well as the absolute numbers of HSCLT, HSCST, and MPP3, were all higher than or significantly higher than those in the solvent control group, but there were still no statistically significant differences among the treatment groups. It was found that the proportion and absolute number of MPP4 in the combined treatment group were significantly higher than those in the solvent control group, while there were no statistically significant differences between the individual treatment groups and the solvent control group.

[0077] In summary, both pre-irradiation prophylactic administration of estriol and post-irradiation salvage administration of roprostine significantly enhanced the extramedullary hematopoietic activity of the spleen in irradiated mice. It was found that the combined administration of the two drugs significantly enhanced the hematopoietic activity of splenic progenitor cells and some pluripotent progenitor cells such as MPP4.

[0078] Example 5: Combined administration of estriol and rhTPO synergistically protects mice from lethal doses of gamma radiation. This embodiment further explores whether administering estriol for prophylaxis before irradiation and administering rhTPO for treatment after irradiation will produce additive or synergistic protective effects.

[0079] A mouse model of total bodily irradiation (TBI) at a lethal dose of 10.0 Gy was used. The experiment was divided into a solvent control group, an estriol prevention group, an rhTPO treatment group, and an estriol + rhTPO combination treatment group. The solvent control group received the same experimental procedures and total solvent volume as the estriol + rhTPO combination treatment group, administering either estriol or rhTPO solvent at the corresponding administration times. Estriol 5 mg / kg was administered intraperitoneally 48 hours before irradiation; rhTPO 100 µg / kg was administered subcutaneously 30 minutes after irradiation. Figure 8 As shown, mice in the irradiation control and all drug-treated groups experienced similar moderate weight loss within the first 9 days after irradiation. Subsequently, the solvent control group mice experienced progressive weight loss and all died within 14 days after irradiation; the estriol and rhTPO groups experienced transient weight loss from 15 to 30 days after irradiation, while the combined drug-treated groups did not show significant weight loss. Consistent with the weight loss after irradiation, the 30-day survival rates of mice in the estriol, rhTPO, and estriol + rhTPO groups were 70%, 20%, and 90%, respectively. Based on King's formula, the Q value for the combined use of estriol and rhTPO was 1.18, demonstrating a synergistic effect. The above studies indicate that, similar to roprostine, the combined administration of estriol and the thrombopoietin-promoting drug rhTPO can synergistically improve the survival rate of lethal irradiated mice.

[0080] Example 6: Combined administration of estriol and rhTPO significantly promoted the recovery of multilineage hematopoiesis in irradiated mice. To evaluate the effect of combined administration of estriol and rhTPO on hematological recovery in irradiated mice, we compared the effects of estriol and rhTPO alone or in combination on peripheral blood count recovery on days 7, 10, 14, and 20 post-irradiation in a 9.0 Gy whole-body irradiation mouse model. Following 9.0 Gy whole-body irradiation, mice in the solvent control group began to die 9 days post-irradiation, with all mice dying within 22 days. Two mice in the estriol group died during this period. All mice in both the rhTPO monotherapy group and the estriol + rhTPO combination group survived. Figure 9As shown, the peripheral blood counts of mice in the control group and each drug-treated group progressively decreased after irradiation, with various peripheral blood count indicators reaching their lowest points at 10 or 14 days post-irradiation. Subsequently, the peripheral blood counts of surviving mice in each drug-treated group began to recover. The peripheral blood counts of mice in the combined treatment group had essentially recovered to pre-irradiation levels by 14 days post-irradiation, while some indicators in the single-drug groups had not fully recovered to pre-irradiation levels until 20 days post-irradiation. On day 10 post-irradiation, several peripheral blood count indicators in mice treated with estriol and rhTPO alone did not differ significantly from those in the solvent control group. It was found that after combined administration of estriol and rhTPO for prevention and treatment, the number of neutrophils and platelets in peripheral blood was significantly higher than that in the solvent control group and each single-drug group. The platelet count in the combined treatment group was 3.7 times, 4.1 times, and 15.9 times that of the estriol group, rhTPO group, and solvent control group, respectively. Fourteen days after irradiation, peripheral blood counts in the estriol and rhTPO groups showed significant recovery compared to the control group. However, the peripheral blood indicators in the combined treatment group remained significantly higher than those in the single-drug groups. These results indicate that combined administration of estriol and rhTPO significantly promotes the recovery of hematopoiesis in lethally irradiated mice, and its efficacy is significantly superior to that of estriol or rhTPO alone in promoting hematopoietic recovery.

[0081] Example 7: Combined administration of nystatin and roprostine synergistically protects mice from lethal doses of gamma radiation. This embodiment further explores whether administering nystatin before irradiation for prophylaxis and roprostine after irradiation for treatment of acute radiation sickness will produce an additive or synergistic protective effect.

[0082] A mouse model of total bodily exposure (TBI) at a lethal dose of 10.0 Gy was used. The experiment was divided into a solvent control group (PBS), a nystatin prophylaxis group, a roprostine treatment group, and a nystatin + roprostine combination therapy group. The solvent control group received the same experimental procedures and total solvent volume as the nystatin + roprostine combination therapy group, administering either nystatin or roprostine solvent at the corresponding administration times. Nystatin 5 mg / kg was administered intraperitoneally 48 hours before irradiation; roprostine 100 µg / kg was administered subcutaneously 30 minutes after irradiation. Figure 10As shown, mice in the solvent control group experienced a continuous decrease in body weight after irradiation, and all died within 11 days. Mice in the nystatin group reached their lowest body weight 13 days after irradiation, then recovered, with a 30-day survival rate of 40%. Mice in the roprostine group experienced a continuous decrease in body weight throughout the experiment, only recovering 30 days after irradiation, with a 30-day survival rate of only 10%. In contrast, the nystatin + roprostine group maintained a relatively stable body weight from 5 to 30 days after irradiation, with a 30-day survival rate of 90%. Based on the King's Law, the Q-value for the combined use of nystatin and roprostine was 1.96, demonstrating a significant synergistic effect. This further confirms the conclusion that pre-irradiation administration of estrogen-based anti-radiation drugs for prevention and post-irradiation administration of platelet-promoting drugs for treatment of acute radiation sickness have a synergistic effect.

[0083] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. A pharmaceutical composition for preventing and treating acute radiation sickness, characterized in that, The pharmaceutical composition comprises an estrogenic drug and a thrombopoietic drug.

2. The pharmaceutical composition of claim 1, wherein, The estrogenic drug comprises any one or a combination of at least two of estriol, nylestriol, quinestrol, ethinyl estradiol, quinestrol, mestranol, estradiol, estradiol benzoate, estradiol valerate, "523" tablet and / or "500" injection; Preferably, the thrombopoietic drug comprises any one or a combination of at least two of romiplostim, eltrombopag, heptaplostim, avatrombopag, lusfusopag, TPO and rhTPO; Preferably, the estrogenic drug is selected from estriol and / or nylestriol; Preferably, the thrombopoietic drug is selected from romiplostim and / or rhTPO.

3. The pharmaceutical composition of claim 1, wherein, The pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient; Preferably, the pharmaceutically acceptable carrier and / or excipient comprises any one or a combination of at least two of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an emulsifier, a co-solvent, a solubilizer, an osmotic pressure adjusting agent, a surfactant, a coating material, a coloring agent, a pH adjusting agent, an antioxidant, a bacteriostatic agent or a buffer.

4. The pharmaceutical composition of claim 1, wherein, The dose ratio of the estrogenic drug and the thrombopoietic drug is 0.1-500:1; Preferably, the dose ratio of the estrogenic drug and the thrombopoietic drug is 1-300:1; Preferably, the dose ratio of the estrogenic drug and the thrombopoietic drug is 10-200:1; Preferably, the administration mode of the estrogenic drug and the thrombopoietic drug is simultaneous administration or sequential administration; Preferably, the administration mode of the estrogenic drug and the thrombopoietic drug is sequential administration before and after irradiation.

5. The pharmaceutical composition of claim 1, wherein, The dosage form of the pharmaceutical composition comprises a parenteral administration dosage form and a gastrointestinal administration dosage form; Preferably, the parenteral administration dosage form comprises an injection administration dosage form, a respiratory administration dosage form, a cavity administration dosage form, a mucosal administration dosage form, a dermal administration dosage form; Preferably, the gastrointestinal administration dosage form comprises a tablet, a granule, a capsule, a solution, a dry suspension, a powder, a sustained-release preparation, an effervescent tablet, an emulsion, a suspension, a syrup, a drop, a chewable tablet.

6. A kit characterized in that, The kit comprises: (I) a first preparation containing an estrogenic drug; (II) a second preparation containing a thrombopoietic drug.

7. The kit of claim 6, wherein The first preparation is selected from estriol, nylestriol, quinestrol, ethinyl estradiol, quinestrol, mestranol, estradiol, estradiol benzoate, estradiol valerate, "523" tablet and / or "500" injection; Preferably, the first preparation is selected from estriol and / or nylestriol; Preferably, the second preparation is selected from romiplostim, eltrombopag, heptaplostim, avatrombopag, lusfusopag, TPO and / or rhTPO; Preferably, the second preparation is selected from romiplostim and / or rhTPO; Preferably, the kit further comprises an instruction; Preferably, the instruction indicates that the first preparation and the second preparation are used in combination to prevent or / and treat acute radiation sickness.

8. Any of the following uses: (1) The estrogen drug and the thrombopoietic drug in combination for use in preparation of a drug for treating acute radiation sickness; (2) The estrogen drug for use in preparation of a drug for improving the therapeutic effect of the thrombopoietic drug on acute radiation sickness; (3) The thrombopoietic drug for use in preparation of a drug for improving the therapeutic effect of the estrogen drug on acute radiation sickness; (4) The estrogen drug and the thrombopoietic drug in combination for use in preparation of a drug for preventing and / or treating bone marrow suppression caused by radiation exposure; (5) The estrogen drug and the thrombopoietic drug in combination for use in preparation of a drug for preventing and / or treating death of animals or humans caused by radiation exposure; (6) The estrogen drug and the thrombopoietic drug in combination for use in preparation of a drug for preventing and / or treating peripheral blood pancytopenia, leukopenia, neutropenia, lymphopenia, thrombocytopenia, and / or anemia caused by radiation exposure; (7) The estrogen drug and the thrombopoietic drug in combination for use in preparation of a drug for preventing and / or treating damage to hematopoietic stem and / or progenitor cells caused by radiation exposure.

9. Use according to claim 8, characterized in that, The estrogen drug includes any one or a combination of at least two of estriol, nylestriol, quinestrol, ethinyl estradiol, quinestradiol, mestranol, estradiol, estradiol benzoate, "523" tablets, and / or "500" injections; Preferably, the thrombopoietic drug includes any one or a combination of at least two of romiplostim, eltrombopag, heptapag, avatrombopag, lusapag, TPO, and rhTPO; Preferably, the estrogen drug is selected from estriol and / or nylestriol; Preferably, the thrombopoietic drug is selected from romiplostim and / or rhTPO.

10. Use according to claim 8, characterized in that, The acute radiation sickness includes bone marrow type acute radiation sickness, intestinal type acute radiation sickness, and brain type acute radiation sickness; Preferably, the radiation exposure includes alpha radiation exposure, beta radiation exposure, gamma radiation exposure, neutron exposure, proton exposure, heavy ion exposure, electron exposure, and / or X-ray exposure.

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