Application of semiliquidambar cathayensis in preparation of medicine for preventing and treating lung diseases

Abstract

The invention discloses application of semiliquidambar cathayensis in preparation of a medicine for preventing and treating lung diseases, and belongs to the technical field of traditional Chinese medicines. The application refers to application of the semiliquidambar cathayensis, the extract thereof or the pharmaceutical composition thereof in preparation of drugs for preventing and treating pulmonary fibrosis, drugs for preventing and treating lung diseases by improving pulmonary fibrosis and drugs for preventing and treating lung diseases. Tests prove that the semiliquidambar cathayensis can improve the pathological process of pulmonary fibrosis, a new way is provided for treatment of lung diseases, and meanwhile, the application field of the semiliquidambar cathayensis is expanded.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and relates to the application of Semiliquidambar cathayensis H.T.Chang. in the preparation of drugs for preventing and treating lung diseases. Specifically, it relates to the application of Semiliquidambar cathayensis H.T.Chang., its extract or its pharmaceutical composition in the preparation of drugs for preventing and treating lung diseases. Background Art

[0002] Pulmonary fibrosis (PF) is a chronic, progressive, and irreversible interstitial lung disease, which is the common outcome of various types of diffuse interstitial lung diseases. The etiology of PF is complex and its pathogenesis is not yet clear. It is considered to be related to factors such as environmental exposure, smoking, infection, genetic susceptibility, and aging. At present, there is a lack of effective therapeutic drugs for PF. Although pirfenidone and nintedanib can improve clinical symptoms to a certain extent and slow down the decline of lung function, there are certain side effects and drug tolerance problems. Lung transplantation is the only cure for PF and can only be used as a treatment option for a small number of patients. Therefore, strengthening the research and development of new drugs for PF is an urgent problem to be solved in the medical field at present.

[0003] Semiliquidambar cathayensis H.T.Chang. is a plant of the genus Semiliquidambar in the family Hamamelidaceae, and is mainly distributed in the southern and southeastern mountainous areas of China, such as Guangxi, Guangdong, Guizhou and other places. It has the effects of dispelling wind and dampness, relaxing tendons and activating blood circulation, and can be used to treat rheumatic arthritis, traumatic injury, postpartum wind paralysis and other diseases. Modern pharmacological studies have shown that Semiliquidambar cathayensis H.T.Chang. has analgesic, anti-inflammatory, antioxidant, and promoting blood circulation and removing blood stasis effects.

[0004] At present, there are many studies on the efficacy of Semiliquidambar cathayensis H.T.Chang.. CN118021857A discloses the application of Semiliquidambar cathayensis H.T.Chang. in the preparation of drugs for preventing and / or treating liver injury, and CN118436695A discloses the application of Semiliquidambar cathayensis H.T.Chang. in the preparation of anti-pancreatic cancer drugs. Sun Jing et al. (Pharmacological activity study of Semiliquidambar cathayensis H.T.Chang. against hepatitis B virus) disclosed that Semiliquidambar cathayensis H.T.Chang. has the activity of anti-hepatitis virus. After retrieval, no research on the prevention and treatment of pulmonary fibrosis by Semiliquidambar cathayensis H.T.Chang. has been found yet. Summary of the Invention

[0005] The purpose of the present invention is to provide the application of Semiliquidambar cathayensis H.T.Chang. in the preparation of drugs for preventing and treating lung diseases; in particular, the application of Semiliquidambar cathayensis H.T.Chang., its extract or its pharmaceutical composition in the preparation of drugs for preventing and treating lung diseases. The present invention discovers that Semiliquidambar cathayensis H.T.Chang. can relieve the progression of pulmonary fibrosis, providing experimental basis and scientific support for the application of Semiliquidambar cathayensis H.T.Chang. in the treatment of lung diseases.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides the use of Semiliquidambar cathayensis H.T.Chang., its extract, or its pharmaceutical composition in the preparation of a medicament for preventing and treating pulmonary fibrosis.

[0008] In a second aspect, the present invention provides the use of Semiliquidambar cathayensis H.T.Chang., its extract, or its pharmaceutical composition in the preparation of a medicament for preventing and treating lung diseases by improving pulmonary fibrosis.

[0009] In a third aspect, the present invention provides the use of Semiliquidambar cathayensis H.T.Chang., its extract, or its pharmaceutical composition in the preparation of a medicament for preventing and treating lung diseases.

[0010] In some embodiments, the lung diseases include at least one of chronic obstructive pulmonary disease, chronic interstitial lung disease, pneumoconiosis, lung injury associated with autoimmune diseases, and other lung diseases.

[0011] Preferably, the chronic obstructive pulmonary disease includes chronic bronchitis and / or emphysema.

[0012] Preferably, the chronic interstitial lung disease includes at least one of idiopathic pulmonary fibrosis and secondary interstitial pneumonia; more preferably, the secondary interstitial pneumonia includes viral pneumonia and / or allergic pneumonia.

[0013] Preferably, the lung injury associated with autoimmune diseases includes connective tissue disease and / or vasculitis-related pulmonary fibrosis.

[0014] Preferably, the other lung diseases include at least one of sarcoidosis, organizing pneumonia, and lymphangitis carcinomatosa of the lung.

[0015] The Semiliquidambar cathayensis H.T.Chang. of the present invention is sourced from Semiliquidambar cathayensis H.T.Chang. (Hamamelidaceae). Preferably, the Semiliquidambar cathayensis H.T.Chang. is the above-ground part and / or underground part of Semiliquidambar cathayensis H.T.Chang., the above-ground part includes the stem and / or branches, and the underground part includes the roots and / or rhizomes.

[0016] The Semiliquidambar cathayensis H.T.Chang. extract in the present invention is a water extract and / or alcohol extract of Semiliquidambar cathayensis H.T.Chang.

[0017] In some embodiments, the preparation method of the alcohol extract of Semiliquidambar cathayensis H.T.Chang. is: adding Semiliquidambar cathayensis H.T.Chang. to alcohol for reflux extraction, followed by solid-liquid separation to obtain the extract.

[0018] In some specific embodiments, the volume fraction of the alcohol is 5-95%, the addition amount of the alcohol is 5-30 times the weight of Semiliquidambar cathayensis H.T.Chang.; the extraction is performed at least once, and each extraction is carried out for 0.5-3 h.

[0019] Preferably, the volume fraction of the alcohol is 20 - 80%; more preferably 30 - 75%; still more preferably 50 - 75%; most preferably 70%.

[0020] Preferably, the addition amount of the alcohol is 8 - 20 times the weight of Semiliquidambar cathayensis; more preferably 8 - 15 times.

[0021] Preferably, the number of extraction times is 1 - 5 times, and each time is 0.5 - 2 hours; more preferably, the number of extraction times is 1 - 3 times, and each time is 1.5 hours.

[0022] In some specific embodiments, the alcohol is ethanol and / or methanol; preferably ethanol.

[0023] In some embodiments, the preparation method of the water extract of Semiliquidambar cathayensis is: adding Semiliquidambar cathayensis to water for reflux extraction, and performing solid - liquid separation to obtain it.

[0024] In some specific embodiments, the addition amount of the water is 5 - 30 times the weight of Semiliquidambar cathayensis; the extraction is performed at least 1 time, and each time is 0.5 - 3 h.

[0025] Preferably, the addition amount of the water is 8 - 20 times the weight of Semiliquidambar cathayensis; more preferably 8 - 15 times.

[0026] Preferably, the number of extraction times is 1 - 5 times, and each time is 0.5 - 2 hours; more preferably, the number of extraction times is 1 - 3 times, and each time is 1.5 hours.

[0027] The Semiliquidambar cathayensis extract of the present invention can also be processed by any one of concentration, enrichment, and purification. Preferably, it is concentration.

[0028] The pharmaceutical composition of the present invention comprises Semiliquidambar cathayensis or its extract and a pharmaceutically acceptable excipient.

[0029] In the third aspect, the present invention provides a pharmaceutical composition prepared from the aforementioned Semiliquidambar cathayensis or its extract and a pharmaceutically acceptable excipient.

[0030] In some specific embodiments, the dosage form of the pharmaceutical composition is a granule, tablet, capsule, powder, pill, spray or liquid preparation.

[0031] The beneficial effects of the present invention are as follows:

[0032] Experiments have proved that Semiliquidambar cathayensis can slow down the pathological process of pulmonary fibrosis, providing a new way for the treatment of lung diseases. At the same time, it expands the application field of Semiliquidambar cathayensis. Description of the Drawings

[0033] Figure 1It is a Masson-stained section of mouse lung tissue pathology.

[0034] Figure 2 It is the statistical result of the fibrotic area of mouse lung tissue

[0035] Figure 3 It is the expression result of the mRNA levels of collagen-related proteins α-SMA and Collagen I genes in mouse lung tissue. Detailed implementation manners

[0036] The following specific examples illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0037] Before further describing the specific implementation manners of the present invention, it should be understood that the protection scope of the present invention is not limited to the specific implementation manners described below; it should also be understood that the terms used in the embodiments of the present invention are for describing specific implementation manners, rather than for limiting the protection scope of the present invention.

[0038] When the embodiments give a numerical range, it should be understood that unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0039] "Pulmonary fibrosis" and "pulmonary interstitial fibrosis" in the present invention have the same meaning and can be replaced with each other.

[0040] The purpose of the "solid-liquid separation" described in the present invention is to achieve the separation of the solid phase and the liquid phase, and conventional technical means can be adopted, such as filtration, centrifugation, pressure filtration or purification separation, preferably filtration.

[0041] The "pharmaceutical composition" described in the present invention contains Semiliquidambar cathayensis Hance or its extract and pharmaceutically acceptable excipients. In specific implementation manners, the Semiliquidambar cathayensis Hance or its extract described in the present invention is provided in the pharmaceutical composition in an effective amount (such as a therapeutically effective amount).

[0042] As used herein, a "pharmaceutically acceptable" ingredient is a substance that is suitable for use in humans and / or animals without undue adverse side effects, such as toxicity, irritation, and allergic response, i.e., a reasonable benefit / risk ratio. The "pharmaceutically acceptable excipients" include inert diluents, dispersing agents and / or granulating agents, surfactants and / or emulsifying agents, disintegrants, binders, preservatives, buffering agents, lubricants and / or oils. Excipients (such as cocoa butter and suppository wax), coloring agents, coating agents, sweetening agents, and flavoring agents may also be present in the pharmaceutical compositions described herein.

[0043] The "pharmaceutical composition" described in the present invention can be prepared by any method known in the art of pharmacy. Generally, these preparation methods include associating the Semiliquidambar cathayensis or its extract (i.e., the sole active ingredient) with a carrier or excipient and / or one or more other auxiliary ingredients, and then, if necessary and / or desired, shaping and / or packaging the product into the desired single-dose or multi-dose unit.

[0044] The pharmaceutical compositions of the present invention can be prepared according to known methods, such as those described in the general rules for preparation in the Chinese Pharmacopoeia 2020 Edition, the 16th Edition of the Japanese Pharmacopoeia, the United States Pharmacopoeia, and the 9th Edition of the European Pharmacopoeia. Depending on the dosage form, the pharmaceutical compositions of the present invention can be appropriately administered to a patient.

[0045] The sole active ingredient and pharmaceutically acceptable excipients in the "pharmaceutical composition" described herein will vary depending on the identity, body size, and / or condition of the subject being treated and further depending on the route of administration of the composition. The pharmaceutical composition may contain from 0.1% to 100% (w / w) of the first active ingredient.

[0046] As used in the present invention, "treatment", unless otherwise indicated, means reversing, alleviating the disease or disorder to which the term applies or one or more symptoms of such disease or disorder, inhibiting the progression of the disease or disorder or one or more symptoms thereof, or preventing the disease or disorder or one or more symptoms thereof. The term "treatment" as used in the invention refers to a treatment act as just defined above.

[0047] As used herein, the "effective amount" refers to an amount sufficient to elicit the desired biological response. The effective amount of the active ingredient of the present invention may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health status of the subject. In certain embodiments, the effective amount is a therapeutically effective amount. The effective amount is the amount of the sole active ingredient described in the present invention in a single dose. In certain embodiments, the effective amount is the combined amount of the sole active ingredient described in the present invention in multiple doses.

[0048] As used herein, the "therapeutically effective amount" is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or sufficient to delay or minimize one or more symptoms associated with the condition. The therapeutically effective amount of Semiliquidambar cathayensis Hance and its extract means an amount of the therapeutic agent that provides a therapeutic benefit, alone or in combination with other therapies, in the treatment of a condition. The term "therapeutically effective amount" may encompass an amount that modifies the overall therapy, reduces or avoids the symptoms, signs or causes of a condition, and / or enhances the therapeutic efficacy of another therapeutic agent. In certain embodiments, the therapeutically effective amount is an amount sufficient to treat any disease or condition described.

[0049] The present invention does not limit the source of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are all ordinary commercially available products in the technical field.

[0050] It should be noted that bleomycin sulfate (1.5 - 2.0 units / mg) was purchased from Shanghai Macklin Biochemical Co., Ltd.

[0051] The preparation method of the ethanol extract of Semiliquidambar cathayensis Hance is as follows: Semiliquidambar cathayensis Hance (Lingnan Chinese Herbal Pieces Co., Ltd., batch number: 2209001) and 70% ethanol by volume were mixed at a material-liquid mass ratio of 1:15, loaded into a reflux device, heated and refluxed at 70 °C for extraction twice, with each reflux for 1.5 hours, filtered, the filtrates were combined, and concentrated to a medicinal liquid concentration of 1 g of crude drug / mL by a rotary evaporator.

[0052] The preventive and therapeutic effects of Semiliquidambar cathayensis Hance on pulmonary fibrosis

[0053] 1. Experimental animals

[0054] Forty SPF-grade male KM mice, weighing 38 ± 5 g, were provided by Zhuhai Bestone Biotechnology Co., Ltd. All animal experiments and animal feeding were carried out in accordance with the "Animal Experiment Guide of Guangdong Pharmaceutical University".

[0055] 2. Experimental methods

[0056] The experimental animals were raised in an SPF-grade barrier animal room environment with a fixed temperature (22±2℃), stable humidity (50±10%) and a 12-hour light / dark cycle. After 3 days of adaptive feeding, KM mice were randomly divided into 4 groups with 10 mice in each group according to the absence of significant differences in body weight between groups: blank control group (Con), model group (Mod), low-dose Jinlu Banfenghe group (LSC), and high-dose Jinlu Banfenghe group (HSC). At the beginning of the experiment, except for the Con group, the other groups were given 3 mg / kg of bleomycin sulfate by endotracheal intubation to construct a mouse model of pulmonary fibrosis. The LSC group and the HSC group were given Jinlu Banfenghe liquid 2.275 g / kg / d and 6.825 g / kg / d by gavage every other day, and the Con group and the Mod group were given the same volume of water by gavage, and the operation was continued for 14 days.

[0057] 24 hours after the end of the administration on the 14th day, the mice were weighed and all mice were anesthetized with isoflurane. Blood samples were collected by orbital blood sampling, and the collected blood samples were placed in an ultra-speed low-temperature centrifuge and centrifuged at 4°C, 3500r / min for 15min. The supernatant was collected and stored at -80°C for subsequent testing. After the blood was collected, the lungs, intestines and their contents were quickly peeled off. After the last administration, mice in each group were fasted (but not water) for 12 hours. Subsequently, isoflurane was used for inhalation anesthesia, and blood was collected through the intraorbital vein. After the blood was allowed to stand for 2 hours, it was centrifuged at 3500rpm and 4°C for 15 minutes to separate the upper serum or plasma and store it in a -80°C refrigerator. After blood collection, lung tissue was removed. The lungs were photographed, weighed, embedded and frozen, and the rest of the tissues were embedded and frozen.

[0058] 3. Detection indicators

[0059] 3.1 General indicators

[0060] During the experiment, the animals were weighed every 3 days and the changes in general indicators were observed.

[0061] 3.2 Detection of Collagen I and α-SMA gene mRNA levels by real-time quantitative PCR (RT-qPCR)

[0062] RNA from lung tissues was extracted using the FastPure Complex Tissue / Cell Total RNA Isolation Kit (Novagen, Nanjing). After RNA concentration was determined, the mRNA expression of genes was evaluated using the ABI7500 Real-Time System and One-Step RT-qPCR Kit (Novagen, Nanjing).

[0063] 3.3 Lung tissue morphology observation

[0064] After the fresh lung tissue was removed from the body, it was quickly rinsed several times with PBS buffer and then placed in a test tube containing 4% paraformaldehyde solution for fixation at room temperature for 24 hours. After that, the samples were paraffin-embedded, sectioned, and stained with Masson, and photographed to observe the histopathological changes.

[0065] 3.4 Statistical methods

[0066] The experimental data were processed using Prism Graphpad. The measurement data were expressed as X±s. The independent sample t-test was used for comparison between two groups, and ANOVA was used for comparison among multiple groups. P<0.05 was considered statistically significant.

[0067] 4. Experimental results

[0068] 4.1 Effects of Semiliquidambar cathayensis on the lung tissue morphology of pulmonary fibrosis mice

[0069] The effects of Semiliquidambar cathayensis on the deposition of collagen fibers in the lungs of fibrotic mice were observed and analyzed by Masson staining (as Figure 1 shown). The results under the microscope showed that in the Con group, the alveolar structure was intact, the interalveolar septum was normal, and there was no abnormal inflammatory exudation or fibrotic hyperplasia. Masson staining indicated that there was no abnormal blue staining in the lung interstitium, no inflammatory cell infiltration, and no obvious collagen fiber deposition. In the Mod group, a large number of alveoli collapsed and ruptured, broke and fused, and inflammatory exudation and hyperplasia filled the damaged alveoli to form pulmonary consolidation, and the damaged area >50%; Masson staining showed that the lung interstitium was blue-stained in a large area centered on the bronchi, with disordered structure, thickened septum, and inflammatory cell infiltration, indicating a large amount of collagen fiber hyperplasia and diffuse pulmonary fibrosis.

[0070] Compared with the Mod group, the pathological changes of the lung tissue in each administration group (LSC and HSC groups) improved. The alveolar structure was clearly visible in each administration group, and the degree of alveolar wall thickening and the degree of inflammatory cell infiltration were inversely proportional to the administration dose. Although there was still blue staining centered on the bronchi in the Masson staining of each administration group, the lesion area was reduced compared with the Mod group. The blue-stained areas in the LSC and HSC groups were mainly distributed and limited around the bronchi. There were still scattered alveolar fusions and fibrotic-like lesions in the lung interstitium, but more alveolar structures were retained compared with the Mod group, and the blue fiber deposition in the alveolar septum decreased (as Figure 2 shown).

[0071] 4.2 Semiliquidambar cathayensis reduces the expression of collagen genes related to pulmonary fibrosis

[0072] The mRNA levels of collagen I and α-smooth muscle actin genes were evaluated by RT-qPCR analysis (as Figure 3As shown in [figure], compared with the Con group, the mRNA levels of fibrosis markers (Collagen I and α-SMA) in the lung tissues of mice in the Mod group were significantly increased (P < 0.001). Compared with the Mod group, the mRNA levels of fibrosis markers (Collagen I and α-SMA) in the lung tissues of mice in each drug administration group (LSC and HSC groups) were significantly decreased (P < 0.001). The results showed that after intervention with Semiliquidambar cathayensis, the expression levels of fibrosis markers Collagen I and α-SMA were significantly downregulated in bleomycin-induced pulmonary fibrosis in mice.

[0073] In summary, from the results of lung tissue pathology and molecular biology examinations, it can be seen that Semiliquidambar cathayensis can significantly improve the pathological process of pulmonary interstitial fibrosis.

[0074] The above is a further description of the present invention in combination with specific embodiments, but these embodiments are merely exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solutions of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention.

Claims

1. Use of Semiliquidambar cathayensis Hance, its extract or its pharmaceutical composition in the preparation of a medicament for preventing and treating pulmonary fibrosis.

2. Use of Semiliquidambar cathayensis Hance, its extract or its pharmaceutical composition in the preparation of a medicament for preventing and treating pulmonary diseases by improving pulmonary fibrosis.

3. Use of Semiliquidambar cathayensis Hance, its extract or its pharmaceutical composition in the preparation of a medicament for preventing and treating pulmonary diseases.

4. The application according to claim 2 or 3, characterized in that, The pulmonary diseases include at least one of chronic obstructive pulmonary disease, chronic interstitial lung disease, pneumoconiosis, pulmonary injury associated with autoimmune diseases, and other pulmonary diseases; The chronic obstructive pulmonary disease includes chronic bronchitis and / or emphysema; The chronic interstitial lung disease includes at least one of idiopathic pulmonary fibrosis and secondary interstitial pneumonia; The pulmonary injury associated with autoimmune diseases includes connective tissue disease and / or pulmonary fibrosis associated with vasculitis; The other pulmonary diseases include at least one of sarcoidosis, organizing pneumonia, and lymphangitis carcinomatosa.

5. The application according to claim 4, wherein The secondary interstitial pneumonia includes viral pneumonia and / or allergic pneumonia.

6. The application according to any one of claims 1-4, characterized in that, The extract of Semiliquidambar cathayensis Hance is an aqueous extract and / or an ethanol extract of Semiliquidambar cathayensis Hance.

7. The application according to claim 5, characterized in that, The preparation method of the aqueous extract of Semiliquidambar cathayensis Hance is: adding Semiliquidambar cathayensis Hance to water for reflux extraction, and performing solid-liquid separation to obtain it; The preparation method of the ethanol extract of Semiliquidambar cathayensis Hance is: adding Semiliquidambar cathayensis Hance to ethanol for reflux extraction, and performing solid-liquid separation to obtain it.

8. The application according to claim 6, wherein The volume fraction of the ethanol is 5-95%, and the addition amount of the ethanol is 5-30 times the weight of Semiliquidambar cathayensis Hance; the extraction is performed at least once, and each time is 0.5-3 h.

9. The application according to claim 1, characterized in that, The pharmaceutical composition contains Semiliquidambar cathayensis Hance or its extract and a pharmaceutically acceptable excipient; preferably, the pharmaceutical composition is a granule, tablet, capsule, powder, pill, spray or liquid preparation.

10. A pharmaceutical composition, characterized in that, It is made of Semiliquidambar cathayensis Hance or its extract and a pharmaceutically acceptable excipient.

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