Application of 6-acylamino-alpha-aminocaproic acid in preparation of medicine for preventing and / or treating pulmonary fibrosis

Abstract

The invention belongs to the field of medicines, and particularly relates to application of 6-amido-alpha-aminocaproic acid in preparation of medicines for preventing and / or treating pulmonary fibrosis, and the 6-amido-alpha-aminocaproic acid is a compound with a structure as shown in a formula 1 and pharmaceutically acceptable salt or ester thereof; formula 1; r is alkyl with 1 to 4 carbon atoms. The innovative research shows that the structure of the formula 1 can effectively prevent and treat pulmonary fibrosis.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to the technical field of drugs for preventing and / or treating pulmonary fibrosis. BACKGROUND

[0002] Pulmonary fibrosis is a terminal change of a large class of pulmonary diseases characterized by proliferation of fibroblasts and accumulation of a large amount of extracellular matrix, accompanied by inflammatory damage and destruction of tissue structure, and is one of the four major diseases of respiratory diseases and the most serious pathological state of the lung. Its pathological changes are mostly manifested as initial inflammation of the lower respiratory tract, and damage to alveolar epithelial cells and vascular endothelial cells, accompanied by proliferation of fibroblasts and type II alveolar cells, release of cytokines, deposition of extracellular matrix proteins and collagen, and finally causing lung changes. The alveoli of the lungs of patients with pulmonary fibrosis are gradually replaced by fibrous material, resulting in hardening and thickening of the lung tissue, and gradual loss of the gas exchange capacity of the lungs, leading to dyspnea in patients due to varying degrees of hypoxia, and finally death due to respiratory failure. The causes of pulmonary fibrosis are complex, and the pathogenesis is unknown. The currently available drugs and methods for treating pulmonary fibrosis are very limited, and the efficacy is not satisfactory, and the prognosis is very poor, with a 5-year survival rate of only 50%. The causes of pulmonary fibrosis are diverse, and the known causes include external environment (such as long-term exposure to asbestos, silicon dioxide, coal and radiation, etc.), drugs (such as bleomycin, rituximab, propranolol and furantoin, etc.), genetic susceptibility (such as TERT, TERC and MUC5B genes). In summary, there is an urgent need in the industry for high-efficiency and low-toxicity drugs that can effectively prevent and treat pulmonary fibrosis.

[0003] The symptoms and characteristics of pulmonary fibrosis caused by different pathogenic causes are different, and the treatment methods and principles are different. For drug-induced pulmonary fibrosis, the main reason is the side effects of pulmonary fibrosis caused by drug treatment. There are some related treatment methods for drug-induced pulmonary fibrosis in the prior art. For example, the patent document with publication number WO2025095249A1 discloses the use of CXCL11 to treat pulmonary fibrosis by inducing the depolarization of M2 macrophages. The patent document with publication number WO2024193322A1 discloses a method for significantly improving bleomycin-induced pulmonary fibrosis by inhibiting collagen synthesis through inhibition of the glycine transporter GLYT1. The patent document with publication number WO2024186175A1 discloses a new flavonoid derivative and a method for treating pulmonary fibrosis using the flavonoid derivative. For another example, the patent document with publication number WO2023128680A1 discloses a method for treating bleomycin-induced pulmonary fibrosis using guaiacol and 4', 6, 7-trimethoxyisoflavone.

[0004] In summary, the prior art has some solutions for improving drug-induced pulmonary fibrosis, but the prior art still needs to be further expanded, and the prevention and treatment effect and biological safety still need to be further improved. SUMMARY

[0005] The first object of the present application is to provide an application of 6-amido-alpha-aminocaproic acid in the preparation of a drug for preventing and / or treating pulmonary fibrosis.

[0006] The second object of the present application is to provide a drug for preventing and / or treating pulmonary fibrosis comprising the 6-amido-alpha-aminocaproic acid.

[0007] The application of 6-amido-alpha-aminocaproic acid in the preparation of a drug for preventing and / or treating pulmonary fibrosis, wherein the 6-amido-alpha-aminocaproic acid is a compound having the structure of formula 1 and pharmaceutically acceptable salts or esters thereof.

[0008] Formula 1

[0009] The R is a C1-C4 alkyl group.

[0010] The innovative research of the present application shows that the structure of formula 1 can effectively prevent and treat pulmonary fibrosis.

[0011] In the present application, the formula 1 is a compound of formula 1A.

[0012] Formula 1A.

[0013] In the present application, the pulmonary fibrosis can be at least one of primary pulmonary fibrosis, secondary pulmonary fibrosis, idiopathic pulmonary fibrosis, pulmonary interstitial fibrosis, and drug-induced pulmonary fibrosis.

[0014] Preferably, the drug for preventing and / or treating pulmonary fibrosis is a drug for preventing and / or treating drug-induced pulmonary fibrosis.

[0015] In the present application, the drug-induced pulmonary fibrosis is drug-induced pulmonary fibrosis caused by administration of at least one of bleomycin, rituximab, propranolol, and furantoin.

[0016] The present application also provides a pharmaceutical active composition capable of preventing and / or treating pulmonary fibrosis, comprising an active ingredient A and the formula 1; the active ingredient A is a drug component that induces side effects of drug-induced pulmonary fibrosis.

[0017] This invention demonstrates that, thanks to the excellent preventive and therapeutic effects of Formula 1 on pulmonary fibrosis, it can be combined with certain active ingredient A, which has been reported to cause pulmonary fibrosis side effects. This combination achieves the therapeutic goal of active ingredient A while also reducing its pulmonary fibrosis side effects.

[0018] In this invention, the active ingredient A includes at least one of bleomycin, rituximab, propranolol, and nitrofurantoin.

[0019] In this invention, the active ingredients A and Formula 1 exist in the form of a mixture or exist independently of each other before administration.

[0020] The present invention also provides a medicament for the prevention and / or treatment of pulmonary fibrosis, comprising a pharmaceutically effective amount of 6-amido-α-aminocaproic acid.

[0021] The aforementioned medicament for the prevention and / or treatment of pulmonary fibrosis comprises the pharmaceutically active composition described in this invention.

[0022] The aforementioned drug for the prevention and / or treatment of pulmonary fibrosis includes pharmaceutically acceptable excipients and has a pharmaceutically acceptable dosage form.

[0023] In this invention, conventional pharmaceutical methods can be used to produce clinically acceptable, pharmaceutically acceptable dosage forms, such as tablets, capsules, powders, and injections.

[0024] Beneficial effects

[0025] This invention is the first to apply Formula 1 to research related to pulmonary fibrosis and demonstrates for the first time that Formula 1 can significantly improve pulmonary fibrosis caused by bleomycin. Compared to commonly used symptomatic treatments, Formula 1 does not cause any side effects or dependence in humans and is safe to use.

[0026] Furthermore, this invention has also found that Formula 1 can be used to prevent and / or treat drug-induced pulmonary fibrosis caused by drugs such as bleomycin. Attached Figure Description

[0027] Figure 1 The graphs show the changes in body weight of mice after bleomycin modeling in each group, as well as the results of mouse body weight, lung weight, and lung weight-to-body weight ratio at the time of tissue collection. In the graphs, A represents the body weight of each group; B represents the lung weight of each group; and C represents the lung weight / body weight ratio of each group.

[0028] Figure 2 Pathological images of each group stained with marison.

[0029] Figure 3 Images and quantitative results of SMA and COL1 immunohistochemical staining in the lung tissue of mice in each group.

[0030] Figure 4 The results show the hydroxyproline content in the lung tissue of mice in each group.

[0031] Figure 5 The results show the expression of lung fibrosis-related genes in the lung tissues of mice in each group. Detailed Implementation

[0032] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0034] The term "treatment" in this invention refers to reducing the degree of pulmonary fibrosis, curing pulmonary fibrosis to normalize it, or slowing down the progression of pulmonary fibrosis.

[0035] The reagents and raw materials used in this invention are all commercially available.

[0036] Example 1: Preparation of an animal model of pulmonary fibrosis

[0037] 1.1 Main reagents and experimental animals

[0038] Bleomycin used in the experiment was purchased from Nippon Kayaku, batch number X81040.

[0039] Unless otherwise specified, all compounds used in the experiments were purchased from Sigma.

[0040] The SPF-grade C57BL / 6 mice (male, 6-8 weeks old, 16-18g) used in the experiment were purchased from Henan Skbes Experimental Animal Co., Ltd.

[0041] 1.2 Preparation of animal models of pulmonary fibrosis

[0042] Male C57BL / 6 mice (6-8 weeks old) were fasted overnight, anesthetized with sodium pentobarbital (45 mg / kg, ip), and injected intratracheally with bleomycin (1.5 U / kg) once.

[0043] The specific procedure was as follows: After anesthetizing the mice, they were fixed in a prone position. A cold light source was used to illuminate the neck area of ​​the mice. The right hand used forceps to pull the mouse's tongue outward, while the left hand used forceps to open the mouth as much as possible until the glottis was exposed. Then, under the guidance of a guidewire, a 20G cannula was inserted into the mouse's trachea. Afterward, approximately 50 μL of bleomycin was injected into the trachea using a micro-injection needle. The mouse was then rapidly rotated and held upright for 5 minutes to ensure that the bleomycin was evenly distributed into both lobes of the lungs. The entire procedure was performed on an operating table at approximately 60°C.

[0044] Example 2: Treatment and efficacy analysis of pulmonary fibrosis in mice

[0045] 2.1 Formula 1A treatment of bleomycin-induced pulmonary fibrosis in mice

[0046] The specific plan is as follows: After successful modeling, the animal models prepared in Example 1 are administered drugs in groups, and the grouping and drug administration details are shown in Table 1:

[0047]

[0048] 2.2 Analysis of body weight, lung weight, and lung weight-to-body weight ratio of mice in each group

[0049] Seven days after bleomycin induction, mice were administered gavage starting on day 8 and continued for 21 days. During this period, changes in body weight were routinely recorded for each group. After 21 days of gavage, the mice were euthanized, and samples were collected, along with body weight and lung weight data. The lung-to-body weight ratio (LVR) is the ratio of a mouse's lung weight to its total body weight, and is generally used to study the physiological and pathological state of the mouse lungs. The LVR is commonly used as an indicator to assess the health and functional status of the mouse lungs. Figure 1 The results showed that the lung-to-body ratio of mice decreased significantly after treatment with Formula 1A or nintedanib by gavage.

[0050] 2.3 Pathological imaging analysis of masson staining in mice with pulmonary fibrosis

[0051] Masson staining is a classic and authoritative method for staining collagen fibers. After staining, muscle fibers appear red, while collagen fibers appear blue, which is the main way to distinguish between muscle fibers and collagen fibers.

[0052] Right lower lobe lung tissue was collected from the animal, fixed in 4% paraformaldehyde, and embedded in paraffin. Sections were prepared from the largest surface of the paraffin-embedded lung tissue block, and musson staining was performed to observe the fibrosis status. High-resolution pathological images (400x magnification) stained with musson stain were obtained using the Spot Advanced 3.0 high-resolution color pathological image analysis system. The results are shown below. Figure 2 The results showed that bleomycin administration significantly increased the area of ​​blue collagen fibers in the lung tissue of mice, while treatment with Formula 1A or nintedanib significantly reduced the area of ​​blue collagen fibers in the lung tissue of mice.

[0053] 2.4 Immunohistochemical staining and pathological imaging analysis of mice with pulmonary fibrosis

[0054] Immunohistochemical staining utilizes the principle of antigen-antibody reaction. It is a method of determining the location, qualitative and relative quantification of fibrosis-related proteins in tissues by using a chemical reaction to make a chromogenic agent containing labeled antibodies develop color.

[0055] Mouse lung tissue was obtained, fixed in 4% paraformaldehyde, and embedded in paraffin. Sections were prepared from the largest cross-section of the paraffin-embedded liver tissue block. The sections were dewaxed and hydrated, followed by antigen retrieval, endogenous peroxidase inhibition, membrane perforation, and blocking. Primary antibodies (SMA, COL1) were incubated overnight at °C. The next day, secondary antibody incubation, DAB staining, hematoxylin staining, and running water blue staining were performed. After soaking in xylene, the sections were mounted, and the staining of different groups was observed under a microscope. Results are shown in [Figure number missing]. Figure 3 The results showed that bleomycin administration significantly increased the expression of SMA and COL1 in mouse lung tissue, while treatment with Formula 1A or nintedanib significantly reduced the expression of SMA and COL1 in mouse lung tissue.

[0056] 2.5 Determination of hydroxyproline content in pulmonary fibrosis mice

[0057] Hydroxyproline accounts for 13.4% of collagen, a very small amount in elastin, and is absent in other proteins. Therefore, hydroxyproline is used to detect collagen content. The hydroxyproline content of the entire left lung lobe of an animal was detected to evaluate the status of pulmonary fibrosis. The specific method is as follows: The entire left lung lobe of the model animal prepared in Example 1 was taken, and the wet weight was recorded. The homogenate was prepared into a 10% tissue homogenate using physiological saline and ultrasonication. Approximately 150 μL of the homogenate supernatant was taken, and 500 μL of alkaline hydrolysis solution was added. After vortexing and mixing, the mixture was treated with alkaline hydrolysis at 120°C and 0.1 kPa for 40 min (the method was slightly modified according to the kit instructions of Nanjing Jiancheng Bioengineering Technology Co., Ltd.). After adjusting the pH and bringing the volume to a final volume, the supernatant was collected after activated carbon treatment. Hydroxyproline was determined according to the instructions (chloramine T method). The results are shown in Table 2 and... Figure 4 As shown in Table 2, the hydroxyproline content in the model group was significantly higher, indicating severe pathological changes in fibrosis. The hydroxyproline content in the lungs of fibrotic mice treated with Formula 1A or nintedanib was significantly reduced.

[0058]

[0059] 2.6 Determination of expression of pulmonary fibrosis-related genes in pulmonary fibrosis mice

[0060] Acta2, FN1, Col1a1, and Col3a1 genes are commonly used genes for assessing pulmonary fibrosis in mice. Increased expression levels of these genes in lung tissue indicate the severity of pulmonary fibrosis.

[0061] Real-time PCR was used to detect the expression of Acta2, FN1, Col1a1, and Col3a1 genes. A suitable amount of lung tissue was collected, ground, and total RNA was extracted using the Trizol method. Chloroform, isopropanol, and 75% anhydrous ethanol were added sequentially to extract RNA, resulting in a gelatinous RNA precipitate. This precipitate was dissolved in RNase-free water, and a small amount was taken for RNA quality testing. The spectrophotometer was zeroed using RNase-free water, and the absorbance values ​​at 260 nm and 280 nm were read to determine the concentration of the RNA solution. The A260 / A280 ratio of the RNA solution represents the RNA purity, and this ratio should be maintained within the range of 1.8 to 2.1. Following the established laboratory procedure, reverse transcription was performed to obtain cDNA. The PCR reaction mixture was prepared according to the Real-time PCR reagent instructions. A certain amount of cDNA was added to primers, and the mixture was then placed on a 384-well plate and analyzed using a PCR instrument to determine the expression levels of Acta2, FN1, Col1a1, and Col3a1. The results are shown in Table 3 and Figure 5 The model group showed significant expression levels of fibrosis-related genes, indicating severe pathological changes in fibrosis. The Formula 1A treatment group showed significantly reduced expression levels of fibrosis-related genes.

[0062]

[0063] Table 3. Formula 1A reduces the expression levels of fibrosis-related genes in lung tissue of mice with pulmonary fibrosis.

[0064] In this experiment, analysis of results through pathological examination, pathological imaging, and other methods revealed that Formula 1A significantly inhibited bleomycin-induced pulmonary fibrosis; it also significantly reduced the content of hydroxyproline and collagen in the lung tissue of mice with pulmonary fibrosis, as well as the expression levels of related fibrosis genes. These experimental results demonstrate that Formula 1A has excellent therapeutic potential in the treatment of pulmonary fibrosis.

[0065] The results of the above embodiments show that Formula 1A of the present invention has a significant effect against pulmonary fibrosis and can be used as an active ingredient in the preparation of drugs for treating pulmonary fibrosis.

[0066] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

The use of 1,6-amido-α-aminohexanoic acid in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis, characterized in that, The 6-amido-α-aminohexanoic acid is a compound having the structure of Formula 1 and its pharmaceutically acceptable salt or ester; Formula 1 The R mentioned is a C1 to C4 alkyl group.

2. The application as described in claim 1, characterized in that, The aforementioned Formula 1 is a compound of Formula 1A; Formula 1A.

3. The application as described in claim 1 or 2, characterized in that, The drugs mentioned for the prevention and / or treatment of pulmonary fibrosis are drugs for the prevention and / or treatment of drug-induced pulmonary fibrosis.

4. The application as described in claim 3, characterized in that, The drug-induced pulmonary fibrosis mentioned refers to drug-induced pulmonary fibrosis caused by administration of at least one of bleomycin, rituximab, propranolol, and nitrofurantoin.

5. A pharmaceutically active composition capable of preventing and / or treating pulmonary fibrosis, characterized in that, It comprises active ingredient A and formula 1 as described in any one of claims 1 to 4; wherein active ingredient A is a pharmaceutical ingredient that has the side effect of inducing drug-induced pulmonary fibrosis.

6. The pharmaceutically active composition for preventing and / or treating pulmonary fibrosis as described in claim 5, characterized in that, The active ingredient A includes at least one of bleomycin, rituximab, propranolol, and nitrofurantoin.

7. The pharmaceutically active composition for preventing and / or treating pulmonary fibrosis as described in claim 5 or 6, characterized in that, The active ingredients A and Formula 1 exist in the form of a mixture or exist independently of each other before administration.

8. A drug for the prevention and / or treatment of pulmonary fibrosis, characterized in that, It contains a pharmaceutically effective amount of 6-amido-α-aminohexanoic acid.

9. The medicament for the prevention and / or treatment of pulmonary fibrosis as described in claim 8, characterized in that, The pharmaceutically active composition comprises any one of claims 5 to 6.

10. The medicament for the prevention and / or treatment of pulmonary fibrosis as described in claim 8 or 9, characterized in that, It contains pharmaceutically acceptable excipients; Preferably, it has a pharmaceutically acceptable dosage form.

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