Use of stachydrine and its derivatives in the preparation of a medicament for preventing and / or treating pulmonary fibrosis

Abstract

The application belongs to the field of medicine, and particularly relates to application of stachydrine and derivatives thereof to preparation of a medicine for preventing and / or treating pulmonary fibrosis. Researches of the application show that stachydrine can effectively prevent and / or treat pulmonary fibrosis.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, and more specifically to the field of preparing a drug for the prevention and / or treatment of pulmonary fibrosis. Background Technology

[0002] Pulmonary fibrosis is a major category of lung diseases characterized by the end-stage changes of fibroblast proliferation and massive extracellular matrix accumulation, accompanied by inflammatory damage and tissue structure destruction. It is one of the four major respiratory diseases and the most severe pathological condition of the lungs. Its pathological changes mostly manifest as initial lower respiratory tract inflammation, as well as damage to alveolar epithelial cells and vascular endothelial cells, accompanied by fibroblast and type II alveolar cell proliferation, cytokine release, and extracellular matrix protein and collagen deposition, ultimately leading to lung changes. In patients with pulmonary fibrosis, the alveoli are gradually replaced by fibrous material, causing the lung tissue to harden and thicken, gradually losing the lung's gas exchange capacity, leading to varying degrees of hypoxia and dyspnea, and finally death from respiratory failure. The etiology of pulmonary fibrosis is complex, and its pathogenesis is unclear. Currently available drugs and methods for treating pulmonary fibrosis are very limited, with unsatisfactory efficacy and extremely poor prognosis; the 5-year survival rate is only 50%. The causes of pulmonary fibrosis are diverse, including environmental factors (such as long-term exposure to asbestos, silica, coal, and radiation), medications (such as bleomycin, rituximab, propranolol, and nitrofurantoin), and genetic susceptibility (such as genes like TERT, TERC, and MUC5B).

[0003] Stachyose (STC) is a natural compound extracted from Chinese medicinal herbs such as Rehmannia glutinosa and plants of the Stachys genus. Current research has confirmed that stachyose can treat diabetes, polycystic ovary syndrome, castration-resistant prostate cancer, etc., but there are currently no reports on the application of stachyose in the prevention or treatment of pulmonary fibrosis. Summary of the Invention

[0004] The purpose of this invention is to provide the use of stachyose and its derivatives in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis.

[0005] The use of stachyose and its derivatives in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis, wherein the stachyose and its derivatives include at least one of stachyose and its pharmaceutically acceptable salts, esters, glycosides, and amides.

[0006] The application described in this invention involves preparing drugs from stachyose and its derivatives to prevent and / or treat pulmonary fibrosis by regulating the gut microbiota.

[0007] In the application described in this invention, the pulmonary fibrosis is primary pulmonary fibrosis and / or secondary pulmonary fibrosis.

[0008] In the application described in this invention, the secondary pulmonary fibrosis is drug-induced pulmonary fibrosis. The drug inducing drug-induced pulmonary fibrosis is at least one selected from rituximab, propranolol, bleomycin, and nitrofurantoin; preferably bleomycin.

[0009] In a preferred application of the present invention, the stachyose and its derivatives are used in combination with drugs that induce pulmonary fibrosis. This not only effectively exerts the therapeutic effect of the drugs, but also effectively reduces the side effects of pulmonary fibrosis.

[0010] The application described in this invention involves combining it with pharmaceutically acceptable excipients to prepare pharmaceutically acceptable dosage forms. These dosage forms include, for example, subcutaneous injection, intradermal injection, intramuscular injection, intraperitoneal injection, intravenous injection, oral administration, and gavage.

[0011] The present invention also provides a pharmaceutically active composition comprising stachyose and its derivatives and pharmaceutical activity A; wherein pharmaceutical activity A is a drug that can induce pulmonary fibrosis (i.e., a drug that has the side effect of pulmonary fibrosis).

[0012] In this invention, the combined use of stachyose and pharmacologically active ingredient A can enhance the therapeutic function of pharmacologically active ingredient A and effectively reduce the side effects of pulmonary fibrosis. In this invention, pharmacologically active ingredient A includes, but is not limited to, at least one of rituximab, propranolol, bleomycin, and nitrofurantoin, preferably bleomycin.

[0013] In this invention, the components of the active composition exist in a mixture or independently. Alternatively, the components may be used together or sequentially as needed.

[0014] The present invention also provides a medicament comprising a pharmaceutically effective amount of the active composition described herein. In this invention, the medicament may also contain pharmaceutically acceptable excipients and acceptable dosage forms.

[0015] 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.

[0016] Beneficial effects

[0017] This invention develops novel indications for the prevention and / or treatment of stachyose and its derivatives in pulmonary fibrosis. Compared to commonly used symptomatic treatments, stachyose is safe to use as it does not cause any side effects or dependence.

[0018] In this invention, stachyose can be combined with other drugs that can induce fibrosis side effects, which can improve treatment and reduce the pulmonary fibrosis side effects that may occur during treatment. Attached Figure Description

[0019] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings, wherein:

[0020] Figure 1 Images of mice stained with masson dye are shown for each group. A represents the control group (normal mice given saline); B represents the stachyose group (normal mice given stachyose); C represents the model group (mice with pulmonary fibrosis given saline); and D represents the bleomycin + stachyose group (mice with pulmonary fibrosis treated with stachyose).

[0021] Figure 2 Results of hydroxyproline content in lung tissue of bleomycin-treated mice that were given saline or stachyose. Detailed Implementation

[0022] 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.

[0023] 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.

[0024] The reagents and raw materials used in this invention are all commercially available.

[0025] Example 1: Preparation of an animal model of pulmonary fibrosis and its treatment

[0026] 1.1 Main reagents and experimental animals

[0027] Bleomycin used in the experiment was purchased from Nippon Kayaku, batch number X81040.

[0028] Unless otherwise specified, all compounds used in the experiments were purchased from Sigma.

[0029] The SPF-grade C57BL / 6 mice (male, 6-8 weeks old, 16-18g) used in the experiment were purchased from Hunan Slack Jingda Experimental Animal Co., Ltd.

[0030] 1.2 Preparation of animal models of pulmonary fibrosis

[0031] 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 U / kg) for a total of 6 times, with each time intervald by 14 days.

[0032] 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 mice's necks. Using forceps in the right hand, the mouse's tongue was pulled outwards, while using forceps in the left hand, the mouth was opened 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. Approximately 50 μL of bleomycin was then injected into the trachea using a micro-injection needle. The mouse was then rapidly rotated and held upright for 5 minutes to ensure even distribution of the bleomycin into both lung lobes. The entire procedure was performed on an operating table at approximately 60°C. The sham-operated group received an equal volume of injectable saline intratracheally.

[0033] 1.3 Stachyose treatment of bleomycin-induced pulmonary fibrosis in mice

[0034] 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:

[0035] Table 1. Grouping and drug administration in animal models of pulmonary fibrosis after model establishment.

[0036]

[0037] Example 2: Pathological Imaging Analysis of Masson Staining

[0038] 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.

[0039] Right lower lobe lung tissue was collected from the animal, fixed in 4% paraformaldehyde, and embedded in paraffin. Sections of the largest cross-section of the embedded lung tissue were prepared and stained with Masson's stain to observe the fibrosis status. High-resolution pathological images (200x magnification) with Masson's special staining were obtained using the SpotAdvanced 3.0 high-resolution color pathological image analysis system. The results are shown in [Figure number missing]. 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 stachyose significantly reduced the area of ​​blue collagen fibers in the lung tissue of mice.

[0040] Example 3: Determination of hydroxyproline content in pulmonary fibrosis mice

[0041] 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. A 10% tissue homogenate was prepared by ultrasonic homogenization with physiological saline. Approximately 150 μL of the homogenate supernatant was taken, and 500 μL of alkaline hydrolysis solution was added. After vortexing, 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 2 As shown in Table 2, compared with the sham-operated group, the bleomycin group had a significantly higher hydroxyproline content, indicating severe fibrotic pathological changes. Stachyose treatment significantly reduced the hydroxyproline content in the lungs of fibrotic mice.

[0042] Table 2. Stachyose reduces hydroxyproline content in lung tissue of mice with pulmonary fibrosis.

[0043] serial number Group Hydroxyproline content (μg / ml protein) p-value A Sham surgery group 8.10±1.50 B Stachyose group 10.26±2.30 0.843 (AvsB) C Bleomycin group 134.15±5.38 0.000 (AvsC) D Bleomycin + Stachyose group 55.55±13.58 0.000(CvsD)

[0044] In this experiment, analysis of results through pathological examination, pathological imaging, and other methods revealed that stachyose significantly inhibited bleomycin-induced pulmonary fibrosis and significantly reduced the content of hydroxyproline and collagen in the lung tissue of mice with pulmonary fibrosis. These experimental results demonstrate that stachyose has excellent therapeutic potential in the treatment of pulmonary fibrosis.

[0045] The results of the above embodiments show that the stachyose 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.

[0046] 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. Use of stachydrine in the preparation of a medicament for preventing and / or treating pulmonary fibrosis, characterized in that, The pulmonary fibrosis is secondary pulmonary fibrosis; the secondary pulmonary fibrosis is drug-induced pulmonary fibrosis. The drug inducing the drug-induced pulmonary fibrosis is at least one of rituximab, propranolol, bleomycin and furantoin.

2. Use according to claim 1, wherein The stachydrine and the pharmaceutically acceptable excipient are combined to prepare a pharmaceutically acceptable dosage form.

3. A pharmaceutical active composition, characterized in that, The stachydrine and the bleomycin are combined.

4. A medicament, characterized by comprising a compound of the formula (I) or a pharmaceutically acceptable salt thereof. The pharmaceutical active composition of claim 3 is used in a pharmaceutically effective amount.

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