Application of forsythin in preparation of medicine for preventing and / or treating obesity complicated with heart failure

Abstract

The invention discloses application of phillyrin in preparation of a medicine for preventing and / or treating obesity complicated with heart failure, relates to the technical field of biological medicines, and compared with normal-weight HFpEF patients, the obesity HFpEF patients are easier to generate hemodynamic disorder, heart function impairment and more obvious clinical symptoms, and the quality of life is obviously reduced. However, the mechanism of obesity-related myocardial injury is not clear, and clinical drug treatment for obesity-related heart failure patients is very deficient. According to the application of the forsythin in preparation of the medicine for preventing and / or treating the obesity combined with the heart failure, experiments prove that the forsythin can effectively reduce the obesity body weight, improve the symptoms of the heart failure, reduce the heart mass and reduce the myocardial hypertrophy condition, and theoretical guidance is provided for preventing and treating the obesity combined with the heart failure.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to the use of forsythoside in the preparation of drugs for the prevention and / or treatment of obesity complicated with heart failure. Background Technology

[0002] Obesity is showing an epidemic trend. The association between obesity and the risk of new-onset heart failure with preserved ejection fraction (HFpEF) is the most significant form of heart failure. Approximately 80% of HFpEF patients are overweight / obese. Compared to normal-weight individuals, the incidence of HFpEF is increased by 38% and 56% in overweight and obese individuals, respectively. Compared to normal-weight HFpEF patients, obese HFpEF patients are more prone to hemodynamic disturbances, impaired cardiac function, and more pronounced clinical symptoms, resulting in a significantly reduced quality of life. However, the mechanisms of obesity-related myocardial injury are not yet clear, and clinical drug treatments for obesity-related heart failure are severely lacking. Therefore, there is an urgent need to develop a drug that can effectively reduce obesity while simultaneously improving the relief of heart failure-related symptoms.

[0003] Forsythia suspensa, a traditional Chinese medicine, has the effects of clearing heat and detoxifying, reducing swelling and dissipating nodules. Forsythoside is the main pharmacologically active component of Forsythia suspensa and is also the quality control indicator component of Forsythia suspensa as specified in the Chinese Pharmacopoeia. Research shows that forsythoside can reduce the body weight of obese mice and improve insulin resistance. However, it is unclear whether forsythoside can improve obesity-related myocardial damage while effectively reducing the body weight of obese mice. Summary of the Invention

[0004] In view of the above-mentioned problems, the present invention aims to overcome the shortcomings of the prior art by providing a drug for the prevention and / or treatment of obesity complicated with heart failure.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The use of forsythosides in the preparation of drugs for the prevention and / or treatment of obesity complicated with heart failure.

[0006] A drug for the prevention and / or treatment of obesity complicated with heart failure, said drug containing forsythoside.

[0007] Furthermore, through the above technical solution, the dosage form of the drug is an oral dosage form.

[0008] Furthermore, through the above technical solutions, the oral dosage form is a tablet, capsule, drop pill, granule, solution, or suspension.

[0009] The beneficial effects of this invention are: experiments have shown that forsythoside can effectively reduce obesity weight, improve heart failure symptoms, reduce heart mass, and reduce myocardial hypertrophy, providing theoretical guidance for the prevention and treatment of obesity complicated with heart failure. Attached Figure Description

[0010] Figure 1 A schematic diagram illustrating the effect of forsythoside on weight changes in obese animals; Figure 2 This is a schematic diagram showing the effect of forsythoside on serum N-terminal pro-brain natriuretic peptide levels in obese animals. Figure 3 A schematic diagram illustrating the effects of forsythoside on cardiac function in obese animals; Figure 4 A schematic diagram illustrating the effect of forsythoside on heart mass in obese animals; Figure 5 A schematic diagram illustrating the effect of forsythoside on cardiac remodeling in obese animals; Figure 6 This is a schematic diagram illustrating the effect of forsythoside on the level of N-terminal pro-brain natriuretic peptide in the serum of TAC mice. Figure 7 This is a schematic diagram illustrating the effect of forsythoside on cardiac function in TAC mice. Figure 8 This is a schematic diagram illustrating the effect of forsythoside on heart weight in TAC mice. Figure 9 This is a schematic diagram illustrating the effect of forsythoside on cardiac remodeling in TAC mice. Detailed Implementation

[0011] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. The components of the embodiments of this application described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0012] The inventors' research found that compared to patients with normal-weight heart failure with heart failure at the post-extension rate (HFpEF), obese HFpEF patients are more prone to hemodynamic disturbances, more significant cardiac function impairment and clinical symptoms, and a marked decline in quality of life. However, the mechanisms of obesity-related myocardial damage are still unclear, and clinical drug treatments for obesity-related heart failure patients are severely lacking. Therefore, there is an urgent need to develop a drug that can effectively reduce obesity while simultaneously improving the relief of heart failure-related symptoms.

[0013] Example 1 See Figures 1-9, This application discloses the use of phillyrin in the preparation of a drug for preventing and / or treating obesity complicated with heart failure. Phillyrin can effectively reduce obesity weight, improve heart failure symptoms, reduce heart mass, and reduce myocardial hypertrophy, providing theoretical guidance for the prevention and treatment of obesity complicated with heart failure.

[0014] A drug for preventing and / or treating obesity complicated with heart failure, which contains phillyrin. The pharmaceutical excipients are selected from any one or several of fillers, binders, suspending agents, wetting agents, solvents or flavoring agents.

[0015] The dosage form of the drug is an oral dosage form. The oral dosage form is tablets, capsules, dripping pills, granules, solutions or suspensions.

[0016] Example 2 1 Materials and Methods 1.1 Research Objects and Experimental Reagents Male C57BL / 6J mice, SPF grade, were purchased from Jiangsu Jicui Yakang Biotechnology Co., Ltd., and the experimental animal production license number was: SCXK (Su) 2023-0009. The experimental animal breeding environment was SPF grade. According to the requirements of the experimental animal breeding environment in China, the breeding environment was kept well ventilated, the room temperature was maintained at 21-23°C, the relative humidity was 45%-55%, the indoor light and dark alternated for 12 hours each, the experimental mice were allowed to eat and drink freely, and they were raised 5 per cage. Phillyrin was purchased from Chengdu Lemeitian Pharmaceutical Technology Co., Ltd., and the N-terminal pro-brain natriuretic peptide (NT-proBNP) ELISA kit was purchased from Shanghai Sangon Biotech Co., Ltd. The high-fat diet with 60% fat content was purchased from Jiangsu Xietong Biology.

[0017] 1.2 Experimental Grouping and Treatments for Each Group The mice were adaptively fed for one week and randomly divided into a normal group, a high-fat group (HFD), and a phillyrin (Phil) group (25 mg / kg), with 8 mice in each group. The mice in the normal group were fed with a normal diet, and the mice in the other groups were fed with a high-fat diet with 60% fat content and allowed to drink water freely. The phillyrin group was suspended with phillyrin and rosiglitazone in a 1% CMC-Na aqueous solution, and each preparation was for 3 days. The mice in each group were administered the corresponding liquid medicine by gavage every day. The mice in the normal group and the high-fat group were administered an equal volume of solvent control, for 17 consecutive weeks.

[0018] 1.3 Detection of Mouse Body Weight Changes The mice in each group were weighed weekly, and the body weight change curve was recorded.

[0019] 1.4 Detection of Serum N-terminal Pro-brain Natriuretic Peptide (NT-proBNP) Level 10 μL of mouse serum was collected and the serum N-terminal pro-brain natriuretic peptide (NT-proBNP) level was detected using an ELISA kit. The procedure was performed according to the kit instructions.

[0020] 1.5 Evaluation of cardiac function in mice using M-mode ultrasound After weighing the mice, administer 2 μL / g of ready-to-use 1.25% aphthol solution intraperitoneally. Once the mice are anesthetized, apply hair removal cream to the chest and upper abdomen of the mice. After two minutes, wipe off the hair removal cream with a tissue, and then thoroughly wipe off the hair removal cream with a tissue dampened with water to prevent the hair removal cream from corroding the mouse probe. Finally, fix the mouse's limbs to the table and apply coupling gel to the sternum of the mouse and the probe.

[0021] Place the probe beside the mouse's sternum, perpendicular to the sternum, with the Mark point pointing towards the mouse's right shoulder at the 10 o'clock position. In B-Mode, fine-tune the probe until the apex of the heart is pointed, at the same level as the outflow tract, and the outflow tract is unobstructed. This indicates that the standard parasternal long axis has been found. Click M-Mode to display an M-mode echocardiogram. Save the video for subsequent data analysis.

[0022] Left ventricular ejection fraction (EF) and fractional shortening (FS) can be measured based on echocardiographic data.

[0023] 1.6 Mouse heart quality testing Seventeen weeks after modeling and drug administration, mice in each group were sacrificed, their hearts were separated, and weighed.

[0024] 1.7 HE staining and WGA staining of mouse hearts The mouse heart was fixed in 4% paraformaldehyde solution and then placed in an embedding cassette. It was then embedded in paraffin and the routine sectioning procedure was completed.

[0025] First, the sections were dewaxed using xylene, followed by hydration with a gradient of ethanol. For hematoxylin and eosin (HE) staining, the sections were stained with hematoxylin for 3 minutes, then washed with water and allowed to regain their blue color, finally rinsing with running water. Next, the sections were immersed in a gradient of 85% and 95% ethanol for 5 minutes each, followed by staining with eosin for 5 minutes. Then, the sections were dehydrated in anhydrous ethanol for 15 minutes each time, repeated twice. Finally, they were cleared in xylene for 15 minutes each time, also repeated twice. Finally, the sections were mounted with neutral resin.

[0026] After completion, the sections were examined under a microscope, and images were acquired and analyzed using ImageJ software. The paraffin sections were then subjected to WGA staining, followed by routine dewaxing to water. The citrate antigen retrieval solution was heated to boiling, and slides were placed in the solution and boiled for 20 minutes. After natural cooling, the sections were removed and rinsed three times with distilled water for 5 minutes each time.

[0027] On a humidified slide, draw circles around the tissue using an immunohistochemical pen, add 100 μL of WGA staining solution (20 μg / mL), and incubate at 37°C in the dark for 1 hour. Observe the staining under a microscope. After incubation, rinse three times with TBST for 5 minutes each time, and mount with anti-fluorescence quenching mounting medium. Take pictures under a laser confocal microscope, acquire images, and analyze them using ImageJ.

[0028] 1.8 Statistical Methods All data are expressed as mean ± SEM. GraphPad Prism 9.0 software was used to analyze the data or perform t-tests, with P < 0.05 considered statistically significant.

[0029] 2. Experimental Results 2.1 Effects of forsythoside on changes in animal body weight like Figure 1 The results showed that, compared with normally fed mice, high-fat diet significantly increased the body weight of mice, while forsythoside significantly reduced the body weight of mice in the HFD group.

[0030] 2.2 Effects of forsythoside on serum N-terminal pro-brain natriuretic peptide levels in animals Serum N-terminal pro-brain natriuretic peptide (NT-proBNP) levels are important biomarkers in the diagnosis and treatment of heart failure. For example... Figure 2 The results showed that, compared with normal mice, high-fat diet significantly increased the serum NT-proBNP level in mice, while forsythoside showed a trend of decreasing the serum NT-proBNP level in HFD mice (P=0.08).

[0031] 2.3 Effects of forsythoside on cardiac function in animals Mouse cardiac M-mode ultrasound results as follows Figure 3 As shown, compared with normal mice, the EF of HFD mice was significantly decreased, while forsythoside significantly increased the EF of HFD mice. Similarly, compared with normal mice, the FS of HFD mice was significantly decreased, while forsythoside significantly increased the FS of HFD mice.

[0032] 2.4 Effects of forsythoside on animal heart quality like Figure 4As shown, compared with normal mice, the heart mass of HFD mice was significantly increased, suggesting that obesity led to cardiac remodeling in mice, while forsythoside could significantly reduce the heart mass of mice in the HFD group.

[0033] 2.5 Effect of forsythoside on cardiac remodeling in animals As Figure 5 Shown by the HE staining results and WGA staining results in a-b, compared with normal mice, the hearts and cardiomyocytes of HFD mice were significantly enlarged, suggesting that obesity led to myocardial hypertrophy, while forsythoside could significantly reduce myocardial hypertrophy in the HFD group of mice.

[0034] Example 3 Therapeutic effect of forsythoside on mice with heart failure induced by transverse aortic constriction (TAC)

[0035] 1 Materials and methods 1.1 Research objects and experimental reagents Male C57BL / 6J mice, SPF grade, were purchased from Jiangsu Jicui Yakang Biotechnology Co., Ltd., and the experimental animal production license number was: SCXK (Su) 2023-0009. The experimental animal breeding environment was SPF grade. According to the requirements of the experimental animal breeding environment in China, the breeding environment was kept well ventilated, the room temperature was maintained at 21-23 °C, the relative humidity was 45%-55%, the indoor light and dark alternated for 12 hours each, the experimental mice were allowed to eat and drink freely, and they were housed at 5 mice per cage. Forsythoside was purchased from Chengdu Lemeitian Pharmaceutical Technology Co., Ltd.

[0036] 1.2 Experimental grouping and treatments for each group This experiment was set up as follows: Sham operation group (Sham) group; heart failure model (Tac) group; low-dose forsythoside treatment (Phil-low) group; high-dose forsythoside treatment (Phil-high) group; captopril positive drug (CAP) group, a total of 5 groups, with 8 mice in each group. The dosage of forsythoside was obtained according to the results of previous drug toxicity tests.

[0037] Sham group: After the mice were anesthetized and the sternum was cut in the same way as the mice in the Tac group, the aortic arch was not ligated and then sutured.

[0038] Tac group: A pressure-overload heart failure model was constructed using minimally invasive transverse aortic constriction (MTAC). The model was established according to the method described in the references (Zaw AM, et al. J Vis Exp. 2017;(121):55293; Wang X, et al. Front Cardiovasc Med. 2023;10:1110032). The specific operation was as follows: anesthesia was performed by intraperitoneal injection of 1.25% 2,2,2-tribromoethanol (0.2 mL / 10g). The skin was cut along the midline of the neck and chest, and the muscles were separated to both sides of the trachea. The sternum was cut about 5 mm to the second rib. The thymus was separated to expose the aortic arch. A 6-0 non-absorbable suture was held with curved ophthalmic forceps and passed through the lower part of the left aortic arch. A self-made 26 G blunt needle was placed above the aortic arch, ligated, and then pulled out to achieve a 0.45 mm diameter stenosis of the aortic arch. The sternum and skin were then sutured in sequence. Analgesia was achieved by intraperitoneal injection of 60 μL meloxicam (20 mL: 40 mg). Studies have confirmed that insulin resistance develops 12 weeks after arterial arch coarctation surgery, indicating successful Tac modeling.

[0039] Phil Low group: The modeling method was the same as that of the Tac group. One week after modeling, forsythoside 10mg / kg was administered by gavage.

[0040] Phil high group: The modeling method was the same as that of Tac group. One week after modeling, forsythoside 20mg / kg was administered by gavage.

[0041] Cap group: The modeling method was the same as that of the Tac group. One week after modeling, the antibody captopril 8 mg / kg was administered by gavage.

[0042] At week 14 of drug administration, surviving mice in each group were sacrificed. Blood samples were collected from the surviving mice in each group for subsequent testing.

[0043] 1.3 Serum N-terminal pro-brain natriuretic peptide (NT-proBNP) levels were detected using the same method as above.

[0044] 1.4 M-mode ultrasound was used to evaluate cardiac function in mice, using the same method as above.

[0045] 1.5 The heart weight of mice was measured using the same method as above.

[0046] 1.6 Perform HE staining and WGA staining of the heart, using the same methods as above.

[0047] 1.7 Statistical Methods All data are expressed as mean ± SEM. GraphPad Prism 9.0 software was used to analyze the data or perform t-tests, with P < 0.05 considered statistically significant.

[0048] 2. Experimental Results 2.1 Effect of forsythoside on serum N-terminal pro-brain natriuretic peptide levels in TAC mice like Figure 6 The results showed that, compared with normal mice, serum N-terminal pro-brain natriuretic peptide (PTP) levels were significantly elevated in TAC mice, while forsythoside (20 mg / kg) and captopril significantly reduced serum N-terminal PTP levels in TAC mice.

[0049] 2.2 Effects of forsythoside on cardiac function in TAC mice like Figure 7 The results showed that, compared with normal mice, TAC mice had significantly decreased EF and FS, while low- and high-dose forsythoside and captopril could significantly improve EF and FS in TAC mice.

[0050] 2.3 Effects of forsythoside on heart weight in TAC mice like Figure 8 The results showed that, compared with normal mice, the hearts of TAC mice were significantly enlarged, and the low- and high-dose groups of forsythoside and captopril could significantly improve the heart quality of TAC mice.

[0051] 2.4 Effects of forsythoside on cardiac remodeling in TAC mice like Figure 9 The HE staining and WGA staining results in ab showed that, compared with normal mice, the heart and cardiomyocytes of TAC mice were significantly enlarged, while the low- and high-dose groups of forsythoside and captopril significantly reduced myocardial hypertrophy in TAC mice.

[0052] The beneficial effects of this invention are: experiments have shown that forsythoside can effectively reduce obesity weight, improve heart failure symptoms, reduce heart mass, and reduce myocardial hypertrophy, providing theoretical guidance for the prevention and treatment of obesity complicated with heart failure.

[0053] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. Application of forsythoside in the preparation of drugs for the prevention and / or treatment of obesity complicated with heart failure.

2. A drug for the prevention and / or treatment of obesity complicated with heart failure, characterized in that, The drug contains forsythoside.

3. The drug according to claim 2, characterized in that, The drug is in the form of an oral dosage form.

4. The drug according to claim 3, characterized in that, The oral dosage form is a tablet, capsule, pellet, granule, solution, or suspension.

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