Use of eleutheroside E in preparation of a medicine for preventing and / or treating pulmonary fibrosis and the medicine

Abstract

The application belongs to the field of pulmonary fibrosis treatment, and particularly relates to application of eleutheroside E in preparation of a drug for preventing and / or treating pulmonary fibrosis and the drug, wherein the pulmonary fibrosis is drug-induced pulmonary fibrosis; and the drug-induced pulmonary fibrosis is induced by at least one of rituximab, propranolol, bleomycin and nitrofurantoin. Researches show that the eleutheroside E can be used as an active ingredient, and can be effectively used for detoxification of the inducing drugs such as bleomycin, and for preventing and / or treating the pulmonary fibrosis induced by the drugs.

Description

Technical Field

[0001] This invention relates to the field of pulmonary fibrosis treatment, specifically to the application of Acanthopanax senticosin E in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis and the pharmaceutical field. Background Technology

[0002] Pulmonary fibrosis is a group of chronic, progressive, and irreversible lung diseases characterized by the progressive destruction of the lung interstitium and alveolar structure, abnormal activation and proliferation of fibroblasts, and excessive deposition of extracellular matrix replacing normal lung tissue. Its pathophysiological process mainly manifests as follows: various pathogenic factors (such as environmental stimuli, infection, autoimmune abnormalities, drug damage, etc.) induce persistent damage to alveolar epithelial cells, thereby triggering abnormal repair responses, inducing resting fibroblasts to transdifferentiate into myofibroblasts, and synthesizing and secreting large amounts of extracellular matrix components such as type I collagen, type III collagen, and fibronectin, ultimately forming diffuse interstitial pulmonary fibrosis, impairing the lung's gas exchange function, causing progressive dyspnea, and in severe cases, leading to respiratory failure and even death. As a chronic lung disease that seriously threatens human health, the 5-year survival rate of patients with idiopathic pulmonary fibrosis is only 20%–30%, lower than that of many common malignant tumors. Currently, clinical treatment options for pulmonary fibrosis are extremely limited. Existing drugs can only slow the progression of the disease to a certain extent and cannot reverse existing pulmonary fibrosis lesions. Clinical treatment needs are far from being met.

[0003] Currently, the main drugs for treating pulmonary fibrosis are glucocorticoids, immunosuppressants, nintedanib, and pirfenidone, but these can only slow down the disease progression and cannot stop or reverse pulmonary fibrosis. In addition, they have the side effect of long-term drug dependence.

[0004] With technological advancements, existing technologies have also developed various biological and chemical treatment methods. For example, Chinese patent document CN110139657A discloses a pulmonary fibrosis treatment agent containing mesenchymal stem cells. Chinese patent document CN107773559A discloses the application of indolinone compounds in the preparation of drugs for the prevention and treatment of pulmonary fibrosis. Chinese patent document CN116173118A discloses the application of a Chinese ginseng extract in the preparation of an anti-pulmonary fibrosis drug.

[0005] In summary, existing technologies disclose a variety of treatment methods, but these methods are difficult to selectively target and inhibit the activation of lung fibroblasts, and are also difficult to effectively achieve precise treatment of pulmonary fibrosis. Existing treatment methods need to be further expanded. Summary of the Invention

[0006] In view of the problems existing in the prior art, the first objective of the present invention is to provide an application of Acanthopanax senticosin E in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis.

[0007] A second objective of this invention is to provide a drug for the prevention and / or treatment of pulmonary fibrosis comprising the eleutheroside E.

[0008] Fibrosis in different locations (different types of fibrosis) exhibits different disease phenotypes, and fibrosis in the same location caused by different etiologies also shows different disease phenotypes and characteristics. Treatment methods for fibrosis with different characteristics are difficult to simply switch between, and the effectiveness of such switching is unpredictable. Addressing the problems in the prevention and treatment of pulmonary fibrosis, especially drug-induced fibrosis, this invention, after in-depth research, provides the following improved solutions:

[0009] The use of Acanthopanax senticosin E in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis, wherein the pulmonary fibrosis is drug-induced pulmonary fibrosis; wherein the drug causing drug-induced pulmonary fibrosis includes at least one of rituximab, propranolol, bleomycin and nitrofurantoin.

[0010] The present invention demonstrates that Acanthopanax senticosin E can be used as an active ingredient to reduce the toxicity of the inducing drug and to prevent and / or treat the incidence and progression of drug-induced pulmonary fibrosis caused by the inducing drug.

[0011] This invention demonstrates that, targeting the tissue characteristics and pathological features of drug-induced pulmonary fibrosis, the aforementioned Acanthopanax senticosus glycoside E can selectively target and inhibit the activation and proliferation of fibroblasts in lung tissue, reducing excessive extracellular matrix deposition and lung tissue remodeling. Simultaneously, it can improve inflammatory infiltration in lung tissue, inhibit the excessive activation of pro-fibrotic cytokines (TGF-β1, IL-6, etc.), block abnormal activation of pro-fibrotic signaling pathways, thereby alleviating alveolar structural damage, interstitial thickening, and collagen deposition, effectively delaying or even reversing the progression of drug-induced pulmonary fibrosis, and improving lung function and survival status.

[0012] The structural formula of Acanthopanax senticosin E is Formula 1A:

[0013] Formula 1A.

[0014] The application described in this invention uses Acanthopanax senticosin E as a toxicity reducer for inducing drugs, and is used to prepare drugs for preventing the incidence of drug-induced pulmonary fibrosis caused by inducing drugs, or for treating the progression of pulmonary fibrosis caused by inducing drugs.

[0015] The application described in this invention combines Acanthopanax senticosin E with pharmaceutically acceptable excipients to prepare a pharmaceutically acceptable dosage form; preferably, the dosage form is an oral preparation or an nebulizer.

[0016] The present invention also provides a medicament for the prevention and / or treatment of pulmonary fibrosis (also referred to as a combination drug), comprising pharmaceutically effective amounts of active ingredient A and active ingredient B, wherein active ingredient A is eleutheroside E; and active ingredient B comprises at least one of rituximab, propranolol, bleomycin, and nitrofurantoin.

[0017] This invention provides a combination drug that, in addition to possessing the efficacy of active ingredient B, can also reduce drug-induced fibrosis caused by active ingredient B, thus achieving dual pharmacological activity of both the efficacy of active ingredient B and low drug-induced pulmonary fibrosis.

[0018] For example, the active ingredient B is bleomycin. Combining it with eleutheroside E can yield a combination drug that has both anti-cancer properties and low drug-induced pulmonary fibrosis toxicity.

[0019] The medicament for the prevention and / or treatment of pulmonary fibrosis according to the present invention further comprises pharmaceutically acceptable excipients; preferably, it has a pharmaceutically acceptable dosage form, preferably an oral formulation or a nebulizer.

[0020] Beneficial effects

[0021] The present invention demonstrates that the eleutheroside E can be used as an active ingredient to effectively reduce the therapeutic toxicity and side effects of active ingredients such as bleomycin that induce pulmonary fibrosis. Attached Figure Description

[0022] Figure 1 The images are pathological images of each group stained with masson in Example 2, where (a) is the sham-operated group, (b) is the bleomycin group, and (c) is the bleomycin + Formula 1A group.

[0023] Figure 2 The graph shows the results of hydroxyproline content in the lung tissue of bleomycin-treated mice in Example 3, which were treated with saline or Formula 1A. Detailed Implementation

[0024] 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.

[0025] The "pulmonary fibrosis" described in this invention refers to conventional pulmonary fibrosis in the art. Pulmonary fibrosis is characterized by pathological changes typical of idiopathic pulmonary fibrosis, caused by various factors. Specifically, it refers to pulmonary fibrosis in humans or animals, and its symptoms include: lung inflammation and decreased lung function caused by pulmonary fibrosis. The causes of pulmonary fibrosis include: pulmonary fibrosis caused by lung injury, pulmonary fibrosis caused by dust, or pulmonary fibrosis caused by drugs, wherein the drug is bleomycin.

[0026] The pulmonary fibrosis mentioned here refers to either primary (specific) pulmonary fibrosis, i.e., pulmonary fibrosis of unknown cause, or secondary pulmonary fibrosis, i.e., pulmonary fibrosis secondary to a pre-existing disease. Pulmonary fibrosis includes decreased lung function, lung inflammation, and lung injury. The diseases mentioned for pulmonary fibrosis include chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis, or interstitial pneumonia.

[0027] 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.

[0028] To make the above-mentioned objects, features and advantages of the present invention more readily understood, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0029] The reagents and raw materials used in this invention are all commercially available.

[0030] Example 1: Preparation of an animal model of pulmonary fibrosis and its treatment

[0031] 1.1: Main reagents and experimental animals:

[0032] 1.1.1: Main reagents:

[0033] Bleomycin was purchased from Nippon Kayaku Co., Ltd., product batch number: X81040; unless otherwise specified, all compounds used in the experiment were purchased from Sigma-Aldrich; sodium pentobarbital, DMSO, PEG300, Tween-80, physiological saline and other oral solvents and excipients were all commercially available analytical grade reagents.

[0034] 1.1.2: Laboratory animals:

[0035] The experimental animals were SPF-grade male C57BL / 6 mice, 6-8 weeks old, weighing 16-18g; the experimental animals were procured by Hunan Silek Jingda Experimental Animal Co., Ltd. All animal experimental procedures were conducted in accordance with laboratory animal ethics guidelines.

[0036] 1.2: Methods for preparing animal models of pulmonary fibrosis:

[0037] 1.2.1: Modeling reagents and dosage:

[0038] The model groups (groups B and C) received a single intratracheal injection of bleomycin at a dose of 1 U / kg and a volume of 50 μL per mouse; the sham-operated group (group A) received an intratracheal injection of an equal volume of sterile saline without bleomycin stimulation.

[0039] 1.2.2: Mold making operation steps:

[0040] Pretreatment before modeling: Male C57BL / 6 mice were fasted overnight (with normal water supply);

[0041] Anesthesia: Anesthesia was administered via intraperitoneal injection of sodium pentobarbital at a dose of 45 mg / kg via intraperitoneal injection (ip).

[0042] Airway exposure and drug administration: Mice were anesthetized and fixed in a prone position, and their necks were irradiated with a cold light source; the mice's tongues were pulled outward with forceps to fully open the mouth and expose the glottis; a 20G cannula was inserted into the mouse's trachea with the help of a guidewire, and 50μL of bleomycin solution (model group) / an equal volume of physiological saline (sham operation group) was injected into the trachea through the cannula with a micro-injection needle.

[0043] Uniform drug distribution: Immediately after administration, rotate the mouse and keep it upright for 5 minutes to ensure that the drug solution evenly penetrates the left and right lung lobes;

[0044] Operating environment adjustment: The operating table temperature is controlled at 37℃ throughout the entire process;

[0045] 1.3: Dosing regimen of Formula 1A in mice with bleomycin-induced pulmonary fibrosis:

[0046] 1.3.1: Waiting period after model making:

[0047] After all mice underwent tracheal instillation to establish the model, they were fed routinely for 7 days to determine that the model was stable. Once the model was confirmed to be successful, drug gavage intervention was initiated.

[0048] 1.3.2: Grouping, number of animals, and dosing details (corresponding to Table 1):

[0049] Table 1 Grouping, dosage, and solvent system of mice with bleomycin-induced pulmonary fibrosis

[0050] 1.3.3: Unified Operating Procedures:

[0051] Frequency of gavage administration: once daily at a fixed time;

[0052] All three groups used the same mixed solvent, except that the active compound 1A was dissolved in the solvent of group C.

[0053] The total volume of liquid administered to each mouse in groups A, B, and C was exactly the same, eliminating the interference of solvent volume differences on the experimental results.

[0054] Operational control instructions: Group A received only saline intratracheal infusion without bleomycin; Groups B and C both received 1 U / kg bleomycin intratracheal infusion for modeling, the only difference being whether or not they received Type 1A drug intervention afterward.

[0055] Example 2: Pathological Analysis of Masson Staining

[0056] 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.

[0057] Right lower lobe lung tissue was collected from each group of animals in Example 1, fixed in 4% paraformaldehyde, and embedded in paraffin. Sections were taken from the largest cross-section of the paraffin-embedded lung tissue and stained with masson's solution to observe the fibrosis status. High-resolution color pathological image analysis system Spot Advanced 3.0 was used to obtain high-resolution pathological images (40x magnification) with masson's special staining. The results are shown in […]. Figure 1 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 significantly reduced the area of ​​blue collagen fibers in the lung tissue of mice.

[0058] Example 3: Determination of hydroxyproline content in pulmonary fibrosis mice

[0059] 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 in the lungs of each group of animals in Example 1 was detected to evaluate the status of pulmonary fibrosis. The specific method is as follows: A portion of the lung lobe of the model animal prepared in Example 1 was taken, the wet weight was recorded, 1 mL of alkaline hydrolysis solution was added, and the mixture was treated with alkaline hydrolysis at 95℃ and 0.1 kPa for 30 min (method 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 then determined according to the instructions. The results are shown in Table 2 and... Figure 2 As shown in Table 2, compared with the sham-operated group, the hydroxyproline content in the bleomycin group was significantly increased, indicating severe pathological changes in fibrosis. Formula 1A treatment significantly reduced the hydroxyproline content in the lungs of fibrotic mice.

[0060] Table 2 Formula 1A reduces hydroxyproline content in lung tissue of mice with pulmonary fibrosis

[0061] Note: P < 0.05 indicates that the difference is statistically significant, and P < 0.01 indicates that the difference is extremely significant.

[0062] In this experiment, pathological examination and other methodological analyses revealed that Formula 1A significantly inhibited bleomycin-induced pulmonary fibrosis and significantly reduced the hydroxyproline and collagen content in the lung tissue of mice with pulmonary fibrosis. These experimental results demonstrate that Formula 1A shows promise for the treatment of drug-induced pulmonary fibrosis.

[0063] The results of the above embodiments show that Formula 1A of the present invention has a significant anti-pulmonary fibrosis effect and can be used as an active ingredient in the preparation of drugs for anti-pulmonary fibrosis.

[0064] 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

1. The use of eleutheroside E in the preparation of drugs for the prevention and / or treatment of drug-induced pulmonary fibrosis, characterized in that, Drugs for the purpose of preventing the incidence of drug-induced pulmonary fibrosis caused by precipitating drugs, or for treating the progression of pulmonary fibrosis caused by precipitating drugs. Bleomycin is a drug that can induce drug-induced pulmonary fibrosis.

2. The application as described in claim 1, characterized in that, Acanthopanax senticosin E, combined with pharmaceutically acceptable excipients, is used to prepare pharmaceutically acceptable dosage forms.

3. The application as described in claim 2, characterized in that, The dosage form is an oral preparation or an nebulizer.

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