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

Abstract

The invention discloses application of semiliquidambar cathayensis in preparation of a medicine for preventing and treating heart 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 myocardial fibrosis, drugs for preventing and treating heart diseases by improving myocardial fibrosis and drugs for preventing and treating heart diseases. Tests prove that the semiliquidambar cathayensis disclosed by the invention has a protection effect on the heart, and can be used for reducing fiber tissue hyperplasia and relieving cardiac fibrosis, so that a basis is provided for clinical medication conversion of the semiliquidambar cathayensis.

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 heart diseases, specifically to the application of Semiliquidambar cathayensis H.T.Chang., its extract or its pharmaceutical composition in the preparation of drugs for preventing and treating heart diseases. Background Art

[0002] Myocardial fibrosis is a pathological change of the heart, caused by the excessive accumulation of collagen fibers in the normal tissue structure of the myocardium, and exists in a variety of cardiovascular diseases. Cardiac fibrosis can cause myocardial structural disorders, increased tissue heterogeneity, etc., which is the structural basis for inducing arrhythmias and a potential risk factor for sudden death and chronic heart failure. The research on the treatment methods of cardiac fibrosis is a current research hotspot.

[0003] Semiliquidambar cathayensis H.T.Chang. is a plant of the genus Semiliquidambar in the family Hamamelidaceae, and is currently 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 muscles and activating blood circulation, and can be used to treat rheumatic arthritis, traumatic injuries, 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 to remove 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 myocardial 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 heart 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 heart diseases. The present invention discovers that Semiliquidambar cathayensis H.T.Chang. can relieve the progression of cardiac fibrosis, providing experimental basis and scientific support for the application of Semiliquidambar cathayensis H.T.Chang. in the treatment of heart diseases.

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

[0007] In the first aspect, the present invention provides the application of Semiliquidambar cathayensis H.T.Chang., its extract or its pharmaceutical composition in the preparation of drugs for preventing and treating myocardial 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 heart diseases by improving myocardial 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 heart diseases.

[0010] In some embodiments, the heart diseases include at least one of hypertensive heart disease, coronary heart disease, myocardial infarction, myocardial hypertrophy, myocardial ischemia-reperfusion injury, chronic heart failure, arrhythmia, and diabetic cardiomyopathy.

[0011] 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.

[0012] The Semiliquidambar cathayensis H.T.Chang. extract in the present invention is a water extract and / or an alcohol extract of Semiliquidambar cathayensis H.T.Chang.

[0013] 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, and performing solid-liquid separation to obtain it.

[0014] 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 time is 0.5-3 h.

[0015] Preferably, the volume fraction of the alcohol is 20-80%; more preferably 30-75%; even more preferably 50-75%; most preferably 70%.

[0016] Preferably, the addition amount of the alcohol is 8-20 times the weight of Semiliquidambar cathayensis H.T.Chang.; more preferably 8-15 times.

[0017] Preferably, the number of extractions is 1-5 times, and each time is 0.5-2 hours; more preferably, the number of extractions is 1-3 times, and each time is 1.5 hours.

[0018] In some specific embodiments, the alcohol is ethanol and / or methanol; preferably ethanol.

[0019] In some embodiments, the preparation method of the water extract of Semiliquidambar cathayensis H.T.Chang. is: adding Semiliquidambar cathayensis H.T.Chang. to water for reflux extraction, and performing solid-liquid separation to obtain it.

[0020] In some specific embodiments, the amount of water added is 5 to 30 times the weight of Semiliquidambar cathayensis Hance; the extraction is performed at least once, for 0.5 to 3 hours each time.

[0021] Preferably, the amount of water added is 8 to 20 times the weight of Semiliquidambar cathayensis Hance; more preferably 8 to 15 times.

[0022] Preferably, the number of extractions is 1 to 5 times, for 0.5 to 2 hours each time; more preferably, the number of extractions is 1 to 3 times, for 1.5 hours each time.

[0023] The Semiliquidambar cathayensis Hance extract of the present invention can also be processed by any one of concentration, enrichment, and purification. Preferably, it is concentrated.

[0024] The pharmaceutical composition of the present invention comprises Semiliquidambar cathayensis Hance or its extract and a pharmaceutically acceptable excipient.

[0025] In a third aspect, the present invention provides a pharmaceutical composition prepared from the aforementioned Semiliquidambar cathayensis Hance or its extract and a pharmaceutically acceptable excipient.

[0026] In some specific embodiments, the dosage form of the pharmaceutical composition is a granule, tablet, capsule, powder, pill, spray, or liquid preparation.

[0027] The beneficial effects of the present invention are as follows:

[0028] (1) Experiments have proven that Semiliquidambar cathayensis Hance has a protective effect on the heart, can reduce fibrous tissue hyperplasia, and alleviate cardiac fibrosis, providing a basis for the clinical drug transformation of Semiliquidambar cathayensis Hance.

[0029] (2) The present invention has first discovered that Semiliquidambar cathayensis Hance has a good therapeutic effect on cardiac fibrosis, providing a new approach for the treatment of heart diseases, and at the same time, expanding the application field of Semiliquidambar cathayensis Hance. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a flowchart of the experimental design for Example 2.

[0031] Figure 2 It is the macroscopic morphology of the mouse heart in Example 2.

[0032] Figure 3 It is the mouse heart weight and heart index of the mice in Example 2.

[0033] Figure 4 It is a Masson staining section of the mouse heart tissue pathology in Example 2.

[0034] Figure 5 It is the statistical result of the fibrosis area of the mouse myocardial tissue in Example 2. ***P < 0.001 vs Con group, # P < 0.05 vs Mod group, ### P < 0.001 vs Mod group.

[0035] Figure 6 For the levels of LDH and cTnT in the serum of the mice in Example 2, *** P < 0.001 vs Con group, ## P < 0.01 vs Mod group, ### P < 0.001 vs Mod group. Detailed implementation manners

[0036] The following uses specific specific examples to 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 specific implementation manners described below; it should also be understood that the terms used in the embodiments of the present invention are used to describe specific specific implementation manners, rather than to limit the protection scope of the present invention.

[0038] When the embodiments give numerical ranges, it should be understood that unless otherwise specified in the present invention, either of the two endpoints of each numerical range and any one 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] "Myocardial fibrosis" and "cardiac 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 used, such as filtration, centrifugation, pressure filtration or column chromatography, etc., and filtration is preferred.

[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, "pharmaceutically acceptable" ingredients are substances that are suitable for use in humans and / or animals without undue adverse side effects (such as toxicity, irritation, and allergic response), i.e., substances having a reasonable benefit / risk ratio. The "pharmaceutically acceptable excipients" include inert diluents, dispersants 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), colorants, coating agents, sweetening agents, and flavoring agents may also be present in the pharmaceutical compositions described herein.

[0043] The "pharmaceutical compositions" 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 Hance 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 desirable, 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 compositions" 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 a 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 action as just defined above.

[0047] As used herein, "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 disorder 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, "therapeutically effective amount" refers to an amount sufficient to provide a therapeutic benefit in the treatment of a disorder or sufficient to delay or minimize one or more symptoms associated with the disorder. The therapeutically effective amount of Semiliquidambar cathayensis Hance or its extract means the amount of the therapeutic agent that provides a therapeutic benefit, alone or in combination with other therapies, in the treatment of a disorder. The term "therapeutically effective amount" may encompass an amount that modifies the overall therapy, reduces or avoids the symptoms, signs, or causes of a disorder, 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 disorder described herein.

[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. It should be noted that the medicinal materials and reagents used in the examples of the present invention are shown in Table 1.

[0050] Table 1 Experimental Medicinal Materials and Reagents

[0051] Name Manufacturer Semiliquidambar cathayensis Hance Sichuan Xingzhikang Biotechnology Co., Ltd. 4% Paraformaldehyde Wuhan Sevier Biotechnology Co., Ltd. Isoflurane Shenzhen Rewod Life Science Co., Ltd. Kit Nanjing Jiancheng Bioengineering Institute

[0052] Example 1 Ethanol Extract of Semiliquidambar cathayensis Hance

[0053] The preparation method is as follows: Semiliquidambar cathayensis Hance (origin: Liuzhou, Guangxi (collected in June 2021), supplier: Sichuan Xingzhikang Biotechnology Co., Ltd.) and 70% ethanol by volume fraction were mixed at a material-liquid mass ratio of 1:15, loaded into a reflux device, heated and refluxed at 70 °C for extraction, extracted 2 times, refluxed for 1.5 hours each time, filtered, the filtrates were combined, and concentrated to a medicinal liquid concentration of 1 g of crude drug / mL by a rotary evaporator.

[0054] Example 2 Prevention and Treatment of Myocardial Fibrosis by Semiliquidambar cathayensis Hance

[0055] 1. Experimental Animals

[0056] 50 SPF-grade male KM mice, weighing 20 - 24 g. Provided by Zhuhai Best Testing Biotechnology Co., Ltd. All animal experiments and the feeding of experimental animals were carried out in accordance with the "Animal Experiment Guide of Guangdong Pharmaceutical University".

[0057] 2. Experimental Methods

[0058] 2.1 Experimental animal grouping

[0059] Fifty male KM mice were used. All the experimental animals were raised in an SPF - level barrier animal house at a fixed temperature of (22 ± 2) °C, a stable humidity of (50 ± 10)%, and a 12 - hour light / dark cycle. After 1 day of adaptive feeding, the KM mice were divided into 5 groups (n = 10) with no significant difference in body weight among groups: sham - operation group (Con group), model group (Mod group), low - dose group of Semiliquidambar cathayensis Hance (LSC), medium - dose group of Semiliquidambar cathayensis Hance (MSC), and high - dose group of Semiliquidambar cathayensis Hance (HSC), with 10 mice in each group. The low - dose group was the equivalent dose group after converting the human drug dosage.

[0060] 2.2 Establishment of myocardial fibrosis model

[0061] The left anterior descending branch (LAD) of the coronary artery of mice was ligated to establish a myocardial infarction model (MI model). Briefly, the mice were anesthetized with 2% isoflurane (company, origin) and fixed in the supine position. Then, the mice were ventilated with a ventilator (Harvard Apparatus), and the chest was opened (respiratory rate, 110 / min). The heart was exposed through a horizontal incision at the 3rd - 4th intercostal muscles on the left sternal margin. The LAD coronary artery was ligated with 7 - 0 surgical sutures, and the chest was closed and sutured. The sham - operation group only passed the thread without ligation. After coronary artery surgical ligation, the color of the anterior wall of the left ventricle turned gray or cyanotic, the wall motion decreased, and the electrocardiogram (ECG) showed ST - segment elevation, indicating the successful establishment of the MI model.

[0062] 2.3 Drug treatment

[0063] In the first week of the experiment, gavage administration was carried out once a day, and the administration continued for 7 days after the model was established; the Con group and the Mod group were gavaged with an equal amount of distilled water, and the other groups were given the corresponding doses of drugs according to the group. The recommended daily dosage of Semiliquidambar cathayensis Hance for humans is 15 g / 60 kg. For mice (50 g), the equivalent dose was 9.1 times the human recommended dose, that is, 2.2755 g / kg.

[0064] In this study, starting from the day of model establishment, the LSC group, MSC group, and HSC group were given Semiliquidambar cathayensis Hance at 113.8 mg / 50 g / d, 341.3 mg / 50 g / d (3 times the clinical dose), and 682.5 mg / 50 g / d (6 times the clinical dose) by gavage, respectively, at a dose of 1 g / mL / d. The mice in the sham - operation group and the myocardial infarction group were given normal saline (0.1 mL / 10 g). All mice were treated at the same time every day for one week. After 7 days of operation, all mice were euthanized after anesthesia with isoflurane.

[0065] The specific experimental design process is as Figure 1 shown.

[0066] 2.4 Sample Collection and Processing

[0067] After the last administration, the mice in each group were fasted (water not restricted) for 12 hours. Subsequently, isoflurane was used for inhalation anesthesia, and blood was collected by intraorbital venous puncture. After the blood was allowed to stand for 2 hours, it was centrifuged at 3500 rpm at 4°C for 15 minutes to separate the upper-layer serum or plasma, which was then stored in a -80°C refrigerator. After blood collection, the heart was excised. The heart was photographed, weighed, embedded, and cryopreserved, and the remaining tissues were all subjected to embedding and cryopreservation.

[0068] 2.5 Enzyme-Linked Immunosorbent Assay (ELISA)

[0069] The blood samples were centrifuged at 3000g for 10 minutes to prepare serum samples. Myocardial injury was evaluated by measuring the levels of serum lactate dehydrogenase (LDH) and cardiac troponin T (cTnT) isoenzymes. ELISA detection was performed using the detection kits provided by Beijing Solarbio Science&Technology Co., Ltd. (China).

[0070] 2.6 Morphological Analysis

[0071] The mouse hearts were isolated and fixed in 4% pre-cooled paraformaldehyde for 48 hours. After paraffin sectioning, Masson staining was used to observe myocardial fibrosis, and images were taken under an optical microscope. Blue staining indicates collagen distribution.

[0072] 2.7 Statistical Analysis

[0073] The experimental data were all expressed as mean ± standard deviation and data analysis was performed using GraphPad Prism statistical software. For experimental data with single-factor multiple-group comparisons, one-way analysis of variance (One-way ANOVA) was used. The statistical significance level was set at p < 0.05, and the difference was considered statistically significant.

[0074] 3. Experimental Results

[0075] 3.1 Heart Morphology and Index

[0076] Figure 2 shows the macroscopic morphology of the mouse hearts. It can be observed that the surface of the hearts of the Con group mice was smooth and the color was normal; on the contrary, the surface of the hearts of the Mod group mice was rough, the left ventricular region was sunken, and there was an infarcted area with a dull color. Figure 3 shows the heart weight and heart index of the mice in each group. Compared with the control group and the Mod group, the heart weight and heart index of the group treated with Semiliquidambar cathayensis Hance decreased, but due to the short time, no statistical significance was shown.

[0077] 3.2 Myocardial tissue pathology

[0078] Figure 4 The Masson staining images of the hearts of mice in each group are shown as follows. The results show that the myocardial tissue structure of mice in the Con group is normal, while significant fibrosis appears in the myocardium of mice in the Mod group, with darker staining and obvious hyperplasia of fibrous tissues such as collagen. The degree of fibrosis in mice in each drug administration group (LSC, MSC, and HSC) is significantly reduced, and the structure of cardiomyocytes is improved, indicating the alleviating effect of administering Semiliquidambar cathayensis Hance on cardiac fibrosis. Quantitative analysis of Masson staining was performed using ImageJ software. As Figure 5 shown, it was found that the fibrotic area in each drug administration group (LSC, MSC, and HSC) was significantly smaller than that in the Mod group (p < 0.05 or p < 0.001), further confirming the alleviating effect of Semiliquidambar cathayensis Hance on cardiac fibrosis.

[0079] 3.3 Serological analysis

[0080] Figure 6 The relevant indicators detected after separating the serum are shown as follows. The results show that compared with the mice in the Mod group, the levels of LDH and cTnT in the serum of mice in each drug administration group (LSC, MSC, and HSC) are significantly reduced (p < 0.01 or p < 0.001), revealing that administering Semiliquidambar cathayensis Hance has a good protective effect on the heart and has a certain effect on improving the continuously progressing cardiac fibrosis injury.

[0081] 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 myocardial fibrosis.

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

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

4. The application according to claim 2 or 3, characterized in that, The heart diseases include at least one of hypertensive heart disease, coronary heart disease, myocardial infarction, myocardial hypertrophy, myocardial ischemia-reperfusion injury, chronic heart failure, arrhythmia and diabetic cardiomyopathy.

5. The application according to any one of claims 1-3, characterized in that, The extract of Semiliquidambar cathayensis Hance is an aqueous extract and / or an ethanol extract of Semiliquidambar cathayensis Hance.

6. The application according to claim 5, wherein 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.

7. The application according to claim 6, characterized in that, The volume fraction of the ethanol is 5-95%, 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.

8. The application according to claim 1, wherein The pharmaceutical composition contains Semiliquidambar cathayensis Hance or its extract and a pharmaceutically acceptable excipient.

9. The application according to claim 8, characterized in that 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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