Application of hydroxynidone in preparation of medicine for preventing and treating radiation-induced pulmonary fibrosis

Abstract

The invention relates to application of hydroxynidone to preparation of a medicine for preventing and / or treating radiation-induced pulmonary fibrosis. Pharmacological experiments prove that the hydroxynidone has a definite resisting effect on radioactive rat lung injury including early inflammatory exudation and pulmonary interstitial fibroblast hyperplasia, and can obviously reduce pulmonary alveolar inflammatory cell exudation and reduce the content of TGF beta 1 in inflammatory exudate; the medium and large doses have the effect of delaying the development of interstitial pneumonia towards pulmonary fibrosis; the lesion degrees of the small, medium and large dosage groups are obviously reduced and are in a dose-effect relationship.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to the use of hydroxynidone in the preparation of medicaments for the prevention and / or treatment of radiation-induced pulmonary fibrosis. Background Technology

[0002] Radiotherapy is a common treatment for patients with malignant lung tumors. While providing therapeutic benefits, it often comes with corresponding side effects. Radiation-induced lung injury is a common complication after radiotherapy for thoracic malignant tumors, with an incidence rate of 5% to 15%. In most cases, patients are asymptomatic, with only lesions appearing on imaging. If the irradiated area is large or the radiation dose is high, or for other reasons, the patient develops infiltrative changes in the lung tissue, resulting in corresponding clinical symptoms such as dyspnea, dry cough, and fever. Clinically, this is what is referred to as radiation-induced "pneumonia," which is the symptomatic stage. In severe cases, it can cause extensive pulmonary fibrosis, respiratory dysfunction, and even respiratory failure.

[0003] Hydroxynidone (code name: F351) is an analogue of pirfenidone. Previous experiments have demonstrated its potential in combating liver fibrosis, but its effects on lung, heart, and kidney diseases remain unclear. Therefore, further research and development are necessary. Summary of the Invention

[0004] In order to overcome the shortcomings of the prior art, the object of the present invention is to provide the use of hydroxynidone in the preparation of a medicament for the prevention and / or treatment of radiation-induced pulmonary fibrosis.

[0005] The objective of this invention is achieved through the following technical solution:

[0006] This invention provides the use of the compound of formula (I) (i.e., hydroxynidone), its solvate, hydrate, prodrug, or pharmaceutically acceptable salt thereof, in the preparation of a medicament for the prevention and / or treatment of radiation-induced pulmonary fibrosis.

[0007]

[0008] According to an embodiment of the present invention, the radiation-induced pulmonary fibrosis is caused by radiotherapy.

[0009] According to an embodiment of the present invention, the radiotherapy utilizes radiation, including α, β, and γ rays produced by radioactive isotopes, and X-rays, electron beams, proton beams, or other particle beams produced by various X-ray therapy machines or accelerators, to treat diseases.

[0010] According to embodiments of the present invention, the pharmaceutically acceptable salt comprises a salt formed by the compound of formula (I) with an organic acid selected from propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, and citric acid; a salt formed by the compound of formula (I) with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; a salt formed by the compound of formula (I) with an inorganic base selected from sodium, potassium, calcium, and aluminum salts; or a salt formed with an organic base, including methylamine salts, ethylamine salts, and ethanolamine. Salt; or salt formed by a compound of formula (I) with an acidic amino acid selected from aspartic acid and glutamic acid, and then with an inorganic base, including sodium, potassium, calcium, aluminum and ammonium salts, or with an organic base, including methylamine, ethylamine and ethanolamine salts; or salt formed by a compound of formula (I) with a basic amino acid selected from lysine, arginine and ornithine, and then with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid and phosphoric acid, or with an organic acid selected from formic acid, acetic acid, picric acid, methanesulfonic acid and ethanesulfonic acid.

[0011] According to an embodiment of the present invention, the pharmaceutically acceptable salt is a salt formed by a compound of formula (I) and an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid and phosphoric acid, or a salt formed by a compound of formula (I) and an inorganic base selected from sodium, potassium, calcium and aluminum salts, or a salt formed by a compound of formula (I) and an organic base, including methylamine salts, ethylamine salts and ethanolamine salts.

[0012] According to embodiments of the present invention, the drug further includes one or more pharmaceutically acceptable carriers or excipients.

[0013] According to an embodiment of the present invention, the drug may be an oral preparation or a parenteral preparation. Further, the oral preparation may be a tablet, capsule, pill, dispersible powder, granule, oral liquid, syrup, elixir, etc.; the parenteral preparation may be an injection, powder for injection, etc.

[0014] According to embodiments of the present invention, the excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, or methylcellulose, etc.

[0015] According to embodiments of the present invention, the excipients may further contain: lubricants such as talc, magnesium stearate, and mineral oil; humectants; emulsifiers and suspending agents; preservatives such as methyl benzoate and hydroxypropyl benzoate; sweeteners and flavoring agents. The medicament can be formulated using methods known in the art to provide immediate, sustained, or delayed release of the active ingredient upon administration to a patient.

[0016] According to an embodiment of the present invention, the drug further includes at least another drug for treating radiation-induced pulmonary fibrosis.

[0017] According to an embodiment of the invention, the drug is administered before or after radiation.

[0018] The present invention also provides a method for preventing and / or treating radiation-induced pulmonary fibrosis, the method comprising administering to a subject a therapeutically effective amount of hydroxynidone, its solvates, hydrates, prodrugs or pharmaceutically acceptable salts thereof, or a composition containing hydroxynidone, its solvates, hydrates, prodrugs or pharmaceutically acceptable salts thereof.

[0019] According to embodiments of the present invention, the hydroxynidone, its solvates, hydrates, prodrugs or pharmaceutically acceptable salts thereof, or compositions containing hydroxynidone, its solvates, hydrates, prodrugs or pharmaceutically acceptable salts thereof, are administered alone or in combination with other drugs for treating radiation-induced pulmonary fibrosis.

[0020] According to an embodiment of the present invention, the radiation-induced pulmonary fibrosis is caused by radiotherapy.

[0021] According to an embodiment of the present invention, the radiotherapy utilizes radiation, including α, β, and γ rays produced by radioactive isotopes, and X-rays, electron beams, proton beams, or other particle beams produced by various X-ray therapy machines or accelerators, to treat diseases.

[0022] According to an embodiment of the present invention, the combined administration may be administered simultaneously or sequentially.

[0023] According to embodiments of the present invention, the hydroxynidone, its solvate, hydrate, drug precursor or pharmaceutically acceptable salt thereof, or a composition containing hydroxynidone, its solvate, hydrate, drug precursor or pharmaceutically acceptable salt thereof, is administered before or after radiation.

[0024] According to an embodiment of the present invention, the dosage of hydroxynidone can be 50-1800 mg / day, for example 80-1700 mg / day, 100-1600 mg / day, 120-1000 mg / day, 180-800 mg / day, 200-600 mg / day, or 250-500 mg / day.

[0025] According to an embodiment of the present invention, the drug can be administered 1-5 times / day, for example 3 times / day.

[0026] According to an embodiment of the present invention, the subject is a mammal, preferably a human.

[0027] The beneficial effects of this invention are as follows: Experiments of this invention demonstrate that F351 has a clear antagonistic effect on radiation-induced lung injury in rats, including early inflammatory exudation and interstitial fibroblast proliferation. It can significantly reduce alveolar inflammatory cell exudation and decrease the TGFβ1 content in inflammatory exudate. At medium and high doses, it can delay the progression of interstitial pneumonia to pulmonary fibrosis. The severity of lesions was significantly reduced in the low, medium, and high dose groups, showing a dose-response relationship. In addition, the experimental results show that the protective effect of the positive control group on lung injury is similar to that of the high dose (no statistical difference). However, it was found in the experiment that the positive control group had significant side effects on the animals. Progressive weight loss, weakness, and cachexia began to appear in the animals after 3 days of treatment. Since the treatment of radiation-induced lung injury requires long-term medication, this positive control group is obviously not suitable for the prevention and treatment of radiation-induced lung injury. In summary, the pharmacological experimental results show that F351 can alleviate radiation-induced pulmonary fibrosis. Attached Figure Description

[0028] Figure 1 Inflammatory cell count in bronchoalveolar lavage fluid of F351 rats. In the figure, *: p<0.05 compared with the model group, **: p<0.01 compared with the model group, #: p<0.05 compared with the previous group, ##: p<0.01 compared with the previous group.

[0029] Figure 2 Protein content in bronchoalveolar lavage fluid of rats in each group of F351. In the figure, *: p<0.05 compared with the model group, #: p<0.05 compared with the previous group, ##: p<0.01 compared with the previous group.

[0030] Figure 3 Determination of TGFβ1 content in bronchoalveolar lavage fluid of F351 rats. In the figure, **: p<0.01 compared with the model group, ##: p<0.01 compared with the previous group.

[0031] Figure 4 The preventive and therapeutic effects of F351 on radiation-induced lung injury (image areal density analysis). In the figure, **: p < 0.01 compared with the model group, ##: p < 0.01 compared with the control group.

[0032] Figure 5 Comparison of lung consolidation images between male and female F351 rats. In the figure, **: p<0.01 between F and M groups.

[0033] Figure 6 : Pathological changes in lung tissue of rats in cage 4 (Ctr F2-4) (Ctr: control group, F: female)

[0034] Figure 7 Image of pathological changes in lung tissue of Rad M5-1 (Rad: irradiated model group, M: male).

[0035] Figure 8 Image of pathological changes in lung tissue of Rad M7-4 (Rad: irradiated model group, M: male).

[0036] Figure 9 Image of pathological changes in lung tissue of Micro M9-4 (Micro: micro group, M: male).

[0037] Figure 10 Image of pathological changes in lung tissue of Mini M15-1 (Mini: low-dose group, M: male).

[0038] Figure 11 Image of lung tissue pathological changes in Mid M19-2 (Mid: medium dose group, M: male).

[0039] Terminology Definitions and Explanations

[0040] The term "subject" in this article refers to an animal, such as a mammal. Mammals include, for example, mice, rats, dogs, cats, pigs, sheep, horses, cattle, and humans.

[0041] The term "therapeutic effective amount" or "effective amount" herein refers to the amount of a compound disclosed and / or described herein that, when administered to a subject requiring this treatment, is sufficient to achieve the treatment as defined herein. A therapeutically effective amount of a compound may be an amount sufficient to treat radiation-induced pulmonary fibrosis. Therapeuticly effective amounts will vary depending on factors such as the subject being treated and their disease condition, the subject's weight and age, the severity of the disease condition, the specific compound, the dosing regimen to be followed, the timing of administration, and the method of administration, all of which can be readily determined by one of ordinary skill in the art.

[0042] The term “treatment” in this article includes one or more of the following: suppressing a disease or condition; slowing or halting the development of clinical symptoms of a disease or condition; and / or alleviating a disease or condition (i.e., causing relief or resolution of clinical symptoms), and a complete or partial reduction of clinical symptoms of a disease or condition.

[0043] The term "therapeutic effect" in this article refers to the effect resulting from treatment, which at the animal level manifests as alteration, usually reduction or improvement of symptoms of disease or disease condition, or cure of disease or disease condition. Detailed Implementation

[0044] The present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0045] Example 1

[0046] I. Materials and Methods

[0047] 1. Tested Drugs

[0048] 1.1 F351 (oxyphenisatinone): provided by Shanghai Ruixing Gene Technology Co., Ltd., off-white crystalline powder, batch number: 050901, purity greater than 99%.

[0049] 1.2 Positive drug: oral dexamethasone hydrochloride, 0.75 mg, 100 tablets / bottle, produced by Zhejiang Xianju Pharmaceutical Co., Ltd., product batch number: 060810.

[0050] 1.3 All tested drugs were ground and dissolved with 0.5% sodium carboxymethylcellulose (produced by Beijing Fengli Jingqiu Trading Co., Ltd.).

[0051] 2. Main Instruments and Detection Kits

[0052] For the determination of total protein in rat bronchoalveolar lavage fluid, a Coomassie Brilliant Blue protein assay kit produced by Nanjing Jiancheng Bioengineering Institute was used. The detection steps were carried out according to the product instruction manual, and a BIO-RAD 550 microplate reader produced in the United States was used for detection; for the determination of TGFβ1 protein in rat bronchoalveolar lavage fluid, a rat TGFβ1 ELISA kit produced by Wuhan Boster Biological Engineering Co., Ltd. was used. The detection steps were carried out according to the product instruction manual, and a BIO-RAD 550 microplate reader produced in the United States was used for detection; the pathological sections and HE staining of rat lung tissues were made by Beijing Xuebang Technology Co., Ltd.; the areal density analysis of pathological images was carried out in the image analysis room of this institute using the true-color pathological tissue image analysis software developed by the General Hospital of the Air Force - Beihang University.

[0053] 3. Tested Animals

[0054] Wistar rats, weighing 160 - 180 g, 15 rats in each group, 8 males and 7 females (the number in individual groups was different, see the section of animal grouping), purchased from the Fengtai Experimental Animal Center of the Academy of Military Medical Sciences, certificate number: SCXK-(Jun)2007 - 004. The rats were raised in the secondary animal laboratory of this hospital, 4 - 5 rats in each cage, fed with standard expanded feed for rats, and allowed free access to water and food. Animal experiment facility use certificate: SYXK-(Jun)2002 - 001.

[0055] 4. Test Methods

[0056] It was carried out by referring to the methods specified in the "Compilation of Guidelines for Preclinical Research of New Drugs (Western Medicines)" and the specific experimental methods in "Pharmacological Experimental Methods" (Third Edition) edited by Xu Shuyun, combined with the professional experience of the research group of this project for more than ten years.

[0057] 4.1 Animal grouping: Rats were randomly divided into 7 groups: normal control (n=15), irradiation model (n=16), micro (n=15), small (n=15), medium (n=15), high dose (n=16), and positive drug control (n=20); odd groups consisted of 8 males and 7 females, and even groups consisted of half males and half females.

[0058] 4.2 Treatment factors: Rats were fixed under anesthesia with sodium pentobarbital (0.03 g / kg). 60 Bilateral chest (lung) irradiation with Coγ rays, dose: 20 Gy, dose rate: 273.8 cGy / min; the patient was kept for 30 days after irradiation.

[0059] 4.3 Drug prevention and treatment: F351 micro, small, medium, and large doses and the positive control group were administered drugs starting 3 days before irradiation and continued for 30 days; the normal control group (Ctr) was given physiological saline; the irradiation model group (Mod) was given solvent (5% sodium carboxymethyl cellulose solvent); drugs were administered once a day, 6 days a week, with a 1-day break between administrations;

[0060] 4.4 Dosage: Microdose group (Mic): 12.5 mg / kg / d; Small dose group (Min): 25 mg / kg / d; Medium dose group (Mid): 50 mg / kg / d; Large dose group (Max): 100 mg / kg / d; Positive control group (Pos): Dexamethasone: Initially 2 mg / kg / d. After administration, the animals in this group were found to be generally emaciated, with negative weight gain. Autopsies of some dead animals revealed gastric mucosal ulcers and bleeding. From day 5, the dose was changed to 1 mg / kg / d. Due to continued weight loss, from day 10, the administration was changed to every other day.

[0061] 4.5 Administration method: Gavage;

[0062] 4.6 Gavage volume: 2 ml / 200g body weight.

[0063] 5. Testing of various indicators

[0064] At that time, 10 or more rats (preferably half male and half female) were randomly selected from each group, anesthetized and fixed on the dissection table, blood was expelled from the abdominal aorta, the right bronchus was clamped after thoracotomy, and the lung tissue was irrigated with 2.5 ml of physiological saline for 4 consecutive times. The irrigating fluid was collected and divided into 3 portions for inflammatory cell counting, protein determination and TGFβ1 determination, respectively. The lower lobe of the right lung was fixed in 10% formaldehyde solution for HE staining section preparation.

[0065] 5.1 Dry / wet weight ratio of lung tissue: Weigh the wet weight of the upper lobe of the right lung, then bake it at 70 degrees for 72 hours, weigh the dry weight, and calculate the wet / dry weight ratio;

[0066] 5.2 Bronchoalveolar lavage fluid inflammatory cell count: The bronchoalveolar lavage fluid was diluted 1:1 with 1% glacial acetic acid buffer and the total number of inflammatory cells was counted under an optical microscope;

[0067] 5.3 Determination of protein leakage in bronchoalveolar lavage fluid: Performed according to the instructions;

[0068] 5.4 Determination of TGFβ content in bronchoalveolar lavage fluid: Performed according to the instructions.

[0069] 6. Pathological observation and analysis

[0070] 6.1 Lesion description and grading: Under an optical microscope, observe, analyze and describe the lesions, classify them pathologically according to their characteristics, take photographs, and compile a lesion statistical list;

[0071] 6.2 Semi-quantitative analysis of image scanning areal density: According to the method reported in foreign literature, 10 parts were randomly selected from each slice for two-dimensional areal density analysis, and the density analysis of the area of ​​lung interstitial consolidation area / area of ​​lung tissue field was performed.

[0072] 7. Data Processing (Statistical Analysis of Differences Between Groups)

[0073] All data are expressed as mean ± standard deviation, and differences between groups were statistically analyzed using Student's t-test.

[0074] II. Test Results

[0075] 1. Statistics on rat mortality in each experimental group

[0076] Near the end of the experiment, some groups experienced individual rat deaths, ranging from 1 to 2 rats. This was a common occurrence and was not directly related to whether or not the drug was administered or the dosage (see Table 1).

[0077] Table 1. Statistics on rat mortality in each experimental group.

[0078]

[0079] 2. Inflammatory cell count in bronchoalveolar lavage fluid from the left lung of a rat

[0080] Bronchoalveolar lavage fluid inflammatory cell counts showed that F351 significantly reduced irradiated inflammatory cell infiltration in the alveolar cavity and exhibited a good dose-response relationship; the high-dose group was more effective than the positive control group (Tables 2 and 3). Figure 1 ).

[0081] Table 2. Inflammatory cell count in bronchoalveolar lavage fluid from the left lung of rats (*10) 4 / ml)

[0082]

[0083] Table 3. P-values ​​for Paired Comparison T-Tests

[0084]

[0085] 3. Protein content in bronchoalveolar lavage fluid

[0086] After irradiation of lung tissue, the exudation of plasma protein components in the alveolar cavity increased significantly. Different doses of F351 could counteract the exudation of plasma proteins to varying degrees, especially the effects of micro and small doses. However, due to the unequal standard deviations, no statistically significant differences were found (Tables 4 and 5). Figure 2 ).

[0087] Table 4. Protein content of bronchoalveolar lavage fluid (g / L)

[0088]

[0089]

[0090] Table 5. P-values ​​for paired t-tests

[0091]

[0092] 4. Determination of TGFβ1 content in bronchoalveolar lavage fluid

[0093] After irradiation, the TGFβ1 content in bronchoalveolar lavage fluid increased significantly. High-dose neutralizing F351 specifically inhibited the increase of TGFβ1 content in bronchoalveolar exudate, showing a highly significant difference compared to the model group; it also showed a highly significant difference compared to the low-dose group; however, its inhibitory effect on TGFβ1 content was slightly weaker than that of the positive control drug (Tables 6 and 7). Figure 3 ).

[0094] Table 6. Determination of TGFβ1 content in bronchoalveolar lavage fluid (pg / ml)

[0095]

[0096] Table 7. P-values ​​of paired t-tests

[0097]

[0098]

[0099] 5. Pathological changes in lung tissue

[0100] 5.1 Control Group (Ctr)

[0101] Male (M): Abundant alveoli, fully open alveolar cavities, thin alveolar septa, no edema or inflammatory cell infiltration observed;

[0102] Female (F): The alveoli are abundant, the alveolar cavities are fully open, the alveolar septa are thin, and normal small blood vessels and small bronchi are visible. No edema or inflammatory cell infiltration is observed.

[0103] 5.2 Model Groups (Mod)

[0104] M: (1) Extensive edema of lung tissue, alveolar cavities filled with exudate and plasma protein, alveolar septa with a few inflammatory cells infiltrating, and a small number of alveolar cavities with compensatory expansion in 2 cases; (2) Scattered or focal alveolar septa widening of lung tissue, with a significant increase in the number of fibroblasts in the widened septa, and alveolar cavities becoming smaller or closed in 4 cases.

[0105] F: (1) Extensive edema of lung tissue, alveolar cavities filled with exudate and plasma protein, widening of alveolar septa accompanied by edema, and compensatory expansion of alveolar cavities in 3 cases; (2) Widening of alveolar septa, with a significant increase in the number of fibroblasts in the widened septa, which fused together to form patches, and alveolar dilation in 4 cases.

[0106] 5.3 Microdose group (Mic)

[0107] M: (1) Extensive edema of lung tissue, alveolar cavities filled with exudate and plasma protein, alveolar septa with a few inflammatory cells infiltrating, and a large number of alveolar cavities with compensatory expansion in 1 case; (2) Local or diffuse alveolar septa widening, with a significant increase in the number of fibroblasts in the septa, and alveolar compensatory expansion in 5 cases.

[0108] F: (1) Pulmonary edema, alveolar cavities filled with exudate and plasma protein, alveolar septa with a few inflammatory cells infiltrating, and more alveolar cavities with compensatory dilation in 2 cases; (2) Diffuse alveolar septal widening, with a significant increase in the number of fibroblasts in the widened septa, which fused together to form patches, and alveolar dilation in 3 cases.

[0109] 5.4 Low-dose group (Min)

[0110] M: (1) Widening of alveolar septa near the pleura, edema of lung tissue, alveolar cavities filled with exudate and plasma protein in 1 case; (2) Diffuse widening of alveolar septa in lung tissue, increased number of fibroblasts, and a large number of foam cells visible in alveolar cavities in 3 cases.

[0111] F: (1) In one case, alveolar septa were widened in some lung tissues and plasma protein exudation was observed in the alveolar cavity; (2) In one case, a few alveolar septa were slightly widened in local lung tissues and no significant increase in fibroblasts was observed; (3) In five cases, alveolar septa were widened near the pleura and the number of fibroblasts increased.

[0112] 5.5 Medium-dose group (Mid)

[0113] M: (1) pulmonary edema, with plasma protein exudation visible in the alveolar cavity in 1 case; (2) widening of alveolar septa, increase in the number of fibroblasts, and significant decrease in the number of alveoli in 4 cases;

[0114] F: (1) Local pulmonary edema, with plasma fluid exudation in the alveolar cavity in 4 cases; (2) Diffuse mild thickening of alveolar septa in local pulmonary tissue, with mild increase in fibroblasts in 1 case.

[0115] 5.6 High-dose group (Max)

[0116] M: (1) Local pulmonary edema, with plasma fluid exudation in the alveolar cavity in 4 cases; (2) Widening of alveolar septa, increased number of fibroblasts, and extensive alveolar cavity expansion in 3 cases.

[0117] F: (1) Mild widening of alveolar septa, increased number of fibroblasts, and significantly reduced number of alveoli in 4 cases; (2) Mild thickening of alveolar septa, with no significant reduction in number of alveoli in 2 cases.

[0118] 5.7 Positive drug group (Pos)

[0119] M: (1) Abundant alveoli, fully open alveolar cavities, and thin alveolar septa in 2 cases; (2) Diffuse mild thickening of alveolar septa in local lung tissue, with an increased number of fibroblasts in 1 case; (3) Scattered mild thickening of alveolar septa in lung tissue, with foam cells in some alveolar cavities in 3 cases.

[0120] F: (1) A small number of alveolar septa were slightly thickened in the lung tissue, and were scattered in 1 case; (2) Local alveolar septa were widened, the number of fibroblasts increased, and the number of alveoli was not significantly reduced in 4 cases.

[0121] 6. Summary of Pathological Observation

[0122] Table 8. Statistical table of lung tissue lesion types and number of cases in each group (cases)

[0123]

[0124] Ctr: Control group; Mod: Model group; Mic: Micro-dose group; Min: Low-dose group; Mid: Medium-dose group;

[0125] Max: High-dose group; Pos: Positive control group.

[0126] 6.1 Number of cases with lesions

[0127] No abnormal changes in lung tissue were observed in the Ctr group; in the Mod group, there were 5 cases of exudative inflammation and 8 cases of interstitial proliferative lesions, totaling 13 cases (the lesions showed cross-contamination, i.e., inflammatory exudation and fibroblast proliferation were present simultaneously on the same slide); in the Mic group, there were 3 cases of exudative inflammation and 8 cases of interstitial proliferative lesions, totaling 11 cases; in the Min group, there were 2 cases of exudative inflammation and 8 cases of interstitial proliferative lesions, totaling 10 cases; in the Mid group, there were 5 cases of exudative inflammation and 5 cases of interstitial proliferative lesions, totaling 10 cases; in the Max group, there were 8 cases of exudative inflammation and 3 cases of interstitial proliferative lesions, totaling 11 cases; in the Pos group, there were 0 cases of exudative inflammation and 5 cases of interstitial proliferative lesions.

[0128] 6.2 Characteristics of the lesions

[0129] The Mod, Mic, and Min groups of animals showed the most cases of pulmonary interstitial fibroblastic proliferative lesions; the Mid group had an equal number of cases of exudative inflammation and fibroblastic lesions; the Max group had far more cases of exudative inflammation than fibroblastic lesions, suggesting that drug F351 has the effect of delaying the progression of interstitial pneumonia to pulmonary fibrosis; the Pos group showed only proliferative lesions and no exudative inflammation, consistent with the characteristic of this positive drug (dexamethasone) to have an anti-inflammatory exudative effect.

[0130] 6.3 Differences in lesions between males and females

[0131] Overall, within each group, the number of cases of exudative inflammation was greater in female animals than in male animals, while the number of cases of proliferative lesions was greater in male animals than in female animals.

[0132] 6.4 Comparison of lesion severity among groups (image areal density analysis)

[0133] After irradiation, the areal density of lung tissue consolidation (inflammatory exudation and interstitial fibroblast proliferation) significantly increased. After treatment with F351, except for the microdose group, all other groups showed some therapeutic effect and exhibited varying degrees of antagonism compared to the model group. The moderate to high dose group showed particularly significant effects, with no statistically significant difference compared to the positive control group (Tables 9 and 10). Figure 4 ).

[0134] Table 9. Areal density analysis of lung injury consolidation images (lesion area / test field area)

[0135]

[0136] Table 10. P-values ​​of paired t-tests

[0137]

[0138] 6.5 Sexual Differences in Aspect Ratio Analysis of Lung Tissue Lesion Images

[0139] Image analysis showed that although the areal density of diseased lung tissue was greater in females than in males in the medium-dose group, and greater in males than in females in the high-dose group, both statistically significant, there was no sex-specific areal density of diseased lung tissue across the groups overall (Tables 11, 12). Figure 5 ).

[0140] Table 11 Comparison of surface density in images of lung consolidation in rats between males and females.

[0141]

[0142]

[0143] *Female data; #Male data.

[0144] Table 12. P-values ​​of paired T-tests for areal density in each group of male and female animals.

[0145]

[0146] Conclusion: F351 has a clear antagonistic effect on radiation-induced lung injury in rats, including early inflammatory exudation and interstitial fibroblast proliferation. It significantly reduces irradiation-induced alveolar inflammatory cell exudation and lowers the TGFβ1 content in alveolar inflammatory exudate. At medium to high doses, F351 delays the progression of interstitial pneumonia to pulmonary fibrosis following lung irradiation. There was no significant difference in the degree of radiation-induced lung lesions between male and female animals. Slight differences in lung lesions were observed between male and female animals within individual groups after administration, but systemically, there was no sex-specific tendency in the severity of lesions. The experiment revealed that the positive control drug had significant side effects on the animals. Progressive weight loss, weakness, and cachexia began to appear in the animals after 3 days of administration. Given that the treatment of radiation-induced lung injury requires long-term administration, this positive control drug is clearly unsuitable for the prevention and treatment of radiation-induced lung injury.

[0147] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Use of the compound represented by formula (I) (i.e., hydroxynidone), its solvate, hydrate, prodrug, or pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of radiation-induced pulmonary fibrosis.

2. The use according to claim 1, characterized in that, The radiation-induced pulmonary fibrosis was caused by radiation therapy.

3. The use according to claim 2, characterized in that, Radiotherapy is the treatment of diseases using radiation, including alpha, beta, and gamma rays produced by radioactive isotopes, and X-rays, electron beams, proton beams, or other particle beams produced by various X-ray therapy machines or accelerators.

4. The use according to claims 1-3, characterized in that, The pharmaceutically acceptable salts include salts formed by the compound of formula (I) with an organic acid selected from propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, and citric acid; or salts formed by the compound of formula (I) with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; or salts formed by the compound of formula (I) with an inorganic base selected from sodium, potassium, calcium, and aluminum salts; or salts formed with an organic base, including methylamine salts, ethylamine salts, and ethanolamine. Salts are salts formed by esterification of acidic amino acids selected from aspartic acid and glutamic acid with inorganic bases, including sodium, potassium, calcium, aluminum, and ammonium salts; or salts formed with organic bases, including methylamine, ethylamine, and ethanolamine salts; or salts formed by esterification of basic amino acids selected from lysine, arginine, and ornithine with inorganic acids selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; or salts formed with organic acids selected from formic acid, acetic acid, picric acid, methanesulfonic acid, and ethanesulfonic acid. Preferably, the pharmaceutically acceptable salt is a salt formed by a compound of formula (I) and an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid and phosphoric acid, or a salt formed by a compound of formula (I) and an inorganic base selected from sodium, potassium, calcium and aluminum salts, or a salt formed by a compound of formula (I) and an organic base, including methylamine salts, ethylamine salts and ethanolamine salts.

5. The use according to claims 1-4, characterized in that, The drug further includes one or more pharmaceutically acceptable carriers or excipients, preferably including lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, or methylcellulose, etc. The excipients may also contain: lubricants such as talc, magnesium stearate, and mineral oil; humectants; emulsifiers and suspending agents; preservatives such as methyl benzoate and hydroxypropyl benzoate; sweeteners and flavoring agents. The drug can be formulated using methods known in the art to provide immediate, sustained, or delayed release of the active ingredient after administration to a patient.

6. The use according to claims 1-5, characterized in that, The drug can be an oral preparation or a parenteral preparation. Further, the oral preparation can be a tablet, capsule, pill, dispersible powder, granule, oral liquid, syrup, elixir, etc.; the parenteral preparation can be an injection, powder injection, etc. Preferably, the drug also includes at least another drug for treating radiation-induced pulmonary fibrosis.

7. A method for preventing and / or treating radiation-induced pulmonary fibrosis, characterized in that, The method comprises administering to a subject a therapeutically effective amount of hydroxynidone, its solvate, hydrate, prodrug, or a pharmaceutically acceptable salt thereof, or a composition containing hydroxynidone, its solvate, hydrate, prodrug, or a pharmaceutically acceptable salt thereof.

8. The use according to claim 7, characterized in that, The radiation-induced pulmonary fibrosis is caused by radiotherapy, which uses radiation, including α, β, and γ rays produced by radioactive isotopes and X-rays, electron beams, proton beams, or other particle beams produced by various X-ray therapy machines or accelerators, to treat the disease. Preferably, the pharmaceutically acceptable salt is a salt formed by hydroxynidone and an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; or a salt formed by hydroxynidone and an inorganic base selected from sodium, potassium, calcium, and aluminum salts; or a salt formed by hydroxynidone and an organic base, including methylamine salts, ethylamine salts, and ethanolamine salts.

9. The use according to claims 7-8, characterized in that, The hydroxynidone, its solvates, hydrates, prodrugs, or pharmaceutically acceptable salts thereof, or compositions containing hydroxynidone, its solvates, hydrates, prodrugs, or pharmaceutically acceptable salts thereof, are administered alone or in combination with other drugs for treating radiation-induced pulmonary fibrosis. Preferably, the combination administration can be simultaneous or sequential. Preferably, the hydroxynidone, its solvates, hydrates, prodrugs, or pharmaceutically acceptable salts thereof, or compositions containing hydroxynidone, its solvates, hydrates, prodrugs, or pharmaceutically acceptable salts thereof, are administered before or after radiation. Preferably, the dosage of hydroxynidone can be 50-1800 mg / day, for example 80-1700 mg / day, 100-1600 mg / day, 120-1000 mg / day, 180-800 mg / day, 200-600 mg / day, or 250-500 mg / day; the frequency of administration can be 1-5 times / day, for example 3 times / day.

10. The use according to claims 7-9, characterized in that, The subjects were mammals, preferably humans.

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