Application of peucedanin in prevention or treatment of hypertrophic cardiomyopathy

Abstract

The invention discloses an application of Nodakenin (NK) in preparation of a medicine for preventing or treating hypertrophic cardiomyopathy and diseases caused by the hypertrophic cardiomyopathy. According to the application disclosed by the invention, the situation that the Nodakenin is used for preventing or treating hypertrophic cardiomyopathy is proposed for the first time, and experiments find that the phenotype of myocardial cell hypertrophy can be obviously inhibited through in-vitro treatment of the Nodakenin; when the Nodakenin-containing feed is used for feeding mice with hereditary hypertrophic cardiomyopathy caused by gene mutation, the Nodakenin is found to obviously relieve pathological cardiac hypertrophy of the mice and improve the cardiac function. Nodakenin can be used for preparing the medicine for resisting the hereditary hypertrophic cardiomyopathy, and a new way and means are provided for treating the hypertrophic cardiomyopathy; in addition, Nodakenin is the most important medicinal component in the traditional Chinese medicinal material umbelliferae plant peucedanum praeruptorum, has high biological safety to organisms, and has a good clinical application prospect.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of nodakenin in the prevention or treatment of hypertrophic cardiomyopathy. Background Technology

[0002] Hypertrophic cardiomyopathy (HCM) is a hereditary myocardial disease characterized by cardiomyocyte hypertrophy, ventricular wall thickening, and myocardial fibrosis. Its occurrence and development are closely related to inflammatory response, oxidative stress, and abnormal deposition of extracellular matrix. There is currently no original and effective drug for hypertrophic cardiomyopathy in China.

[0003] Nodakenin (NK) is a coumarin glycoside compound mainly derived from the roots of plants in the genus *Peucedanum* of the Apiaceae family. It possesses various pharmacological activities, including anti-inflammatory, antioxidant, anti-fibrotic, and immunomodulatory effects. Its chemical formula is: C 20 H 24 O9. Studies have shown that Nodakenin can promote the regeneration of damaged liver by activating farnesol receptors; Nodakenin can inhibit IgE / Ag-mediated activation of basophilic leukemia cells and allergic reactions, exhibiting good in vitro and in vivo anti-inflammatory effects. Currently, there are no publicly available reports on the use of purslane in the treatment of hypertrophic cardiomyopathy. Summary of the Invention

[0004] Purpose of the invention: To address the problems existing in the prior art, this invention provides the application of Nodakenin in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy; this invention is the first to propose that Nodakenin can be used to prepare drugs for the treatment of hypertrophic cardiomyopathy, providing a new approach and means for the treatment of hypertrophic cardiomyopathy.

[0005] The present invention also provides a pharmaceutical composition for the prevention or treatment of hypertrophic cardiomyopathy and the symptoms thereof.

[0006] Technical solution: The application of imperatorin described in this invention in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy, including the application of imperatorin in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy and its related symptoms.

[0007] The hypertrophic cardiomyopathy includes hereditary hypertrophic cardiomyopathy and the symptoms caused by hereditary hypertrophic cardiomyopathy.

[0008] The hypertrophic cardiomyopathy includes pathological myocardial hypertrophy, increased heart weight or left ventricular weight.

[0009] The hypertrophic cardiomyopathy includes an increase in cardiomyocyte area and / or an elevated level of myocardial fibrosis caused by hereditary hypertrophic cardiomyopathy.

[0010] Among them, the application of the purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by inhibiting the increase of cardiomyocyte area.

[0011] The application of the purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy is achieved by reducing the expression of myocardial hypertrophy markers or by inhibiting the hypertrophic phenotype of cardiomyocytes.

[0012] Among them, the use of purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by reducing the weight of the heart or ventricles.

[0013] Among them, the application of the purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by reducing the thickness of the ventricular wall.

[0014] Furthermore, the purslane can be used in the preparation of drugs that inhibit the AngII-induced increase in cardiomyocyte area.

[0015] The present invention relates to a pharmaceutical composition for the prevention or treatment of hypertrophic cardiomyopathy and its associated symptoms, wherein the pharmaceutical composition contains nodakenin as an active ingredient and a pharmaceutically acceptable carrier.

[0016] The dosage form of the pharmaceutical composition is capsule, powder, tablet, granule, pill, injection, syrup, oral liquid, inhaler, ointment, suppository or patch.

[0017] This invention describes the application of Nodakenin to mice with hereditary hypertrophic cardiomyopathy caused by gene mutation. Nodakenin was administered by gavage, and it was found that Nodakenin significantly alleviated pathological myocardial hypertrophy and improved cardiac function in the mice.

[0018] The present invention relates to the use of the pharmaceutical composition described herein in the preparation of remedies for the prevention or treatment of hypertrophic cardiomyopathy and its associated symptoms. This invention discovers a novel medicinal use for nodakenin, which can combat hypertrophic cardiomyopathy and its associated symptoms. Furthermore, nodakenin is the most important medicinal component of the genus *Peucedanum* (Apiaceae family) in traditional Chinese medicine, and is safer for organisms, showing promising clinical application prospects.

[0019] In a hereditary HCM mouse model, this invention demonstrated that Nodakenin significantly improved hypertrophy-related indicators. Compared to the positive control drug metoprolol, Nodakenin showed a similar and significant improvement trend in reducing HW / TL, left ventricular mass, and ventricular wall thickness. The high-dose group showed significantly better improvement in cardiac macroscopicity than the positive control drug. Nodakenin is a major active ingredient in traditional Chinese medicine with good biological sources, and cytotoxicity results also indicate that it has good biocompatibility.

[0020] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0021] 1. This invention utilizes a mouse model of hypertrophic cardiomyopathy with a Myh6 gene R404Q mutation (Myh6R404Q), which is characterized by hereditary myocardial hypertrophy, enlarged heart area, and increased weight of the heart and left ventricle. Nodakenin gavage significantly improved the symptoms of hereditary pathological myocardial hypertrophy in Myh6R404Q mice.

[0022] 2. This invention uses Nodakenin to directly treat hypertrophic cardiomyocytes in vitro, significantly reducing the cardiomyocyte area and decreasing the increase in the expression of deangiotensin II-induced markers of myocardial hypertrophy. In Myh6R404Q mice, Nodakenin was administered by gavage, reducing the weight of the left ventricle and heart, decreasing heart volume, and decreasing diastolic and systolic myocardial wall thickness.

[0023] 3. This invention, through in vitro and in vivo studies, has revealed that Nodakenin possesses significant potential for treating hypertrophic cardiomyopathy and can be developed as a novel anti-hypertrophic cardiomyopathy drug, providing a new approach and method for its treatment. Furthermore, this invention offers entirely new options and ideas for current anti-hypertrophic cardiomyopathy drugs, broadening the selection scope and contributing to the development of this technological field. Attached Figure Description

[0024] Figure 1 The figure shows the area of ​​cardiomyocytes in H9C2 rats after direct in vitro treatment with Nodakenin; **** indicates p<0.0001.

[0025] Figure 2 Figure 1 shows the expression levels of myocardial hypertrophy markers (left: ANP, right: BNP) in H9C2 rat cardiomyocytes after direct in vitro treatment with Nodakenin; * indicates p<0.05; ** indicates p<0.01; **** indicates p<0.0001.

[0026] Figure 3The ratio of heart weight to tibia length and left ventricular weight of Myh6R404Q mice after gavage administration of Nodakenin (left); in the figure, *** indicates p<0.001; **** indicates p<0.0001.

[0027] Figure 4 Images of mouse hearts after Nodakenin gavage administration to Myh6R404Q mice. (Left 1) Heart image of wild-type mouse; (Left 2) Heart image of Myh6R404Q mouse; (Left 3) Heart image of mouse after low-dose (5 mg / kg) Nodakenin gavage administration; (Left 4) Heart image of mouse after high-dose (20 mg / kg) Nodakenin gavage administration; (Left 5) Heart image of mouse after metoprolol (14 mg / kg) gavage administration for hypertrophic cardiomyopathy.

[0028] Figure 5 The images show the results of echocardiography in mice. (Left 1) shows the thickness of the anterior wall of the left ventricle during systole, (Left 2) shows the thickness of the anterior wall of the left ventricle during diastole, (Left 3) shows the thickness of the posterior wall of the left ventricle during systole, and (Left 4) shows the thickness of the posterior wall of the left ventricle during diastole. In the figures, ** indicates p < 0.01; *** indicates p < 0.001, and **** indicates p < 0.0001. Detailed Implementation

[0029] The present invention will be further described below with reference to specific embodiments and accompanying drawings.

[0030] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were purchased from conventional biochemical reagent companies.

[0031] AngII was purchased from MedChemExpress, #HY-13948, CAS No.:4474-91-3.

[0032] Nodakenin was purchased from Bidex Pharmaceuticals (BD636855, CAS 495-31-8).

[0033] Rat cardiomyocytes H9C2 were purchased from Wuhan Pronosei Life Science Technology Co., Ltd., CL-0089.

[0034] Four-week-old wild-type mice (WT mice) and Myh6 gene R404Q point mutant mice (Myh6 R404Q All were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd., including Myh6 gene R404Q point mutant mice (Myh6). R404Q (C57BL / 6JGpt-Myh6) em1Cin(R404Q) / Gpt, Myh6-p.R404Q|StrainNO.T051403. Normal mouse diet was purchased from Jiangsu Xiehe Biotechnology Co., Ltd.

[0035] Example 1

[0036] The effect of Nodakenin on cell area during direct in vitro treatment of cardiomyocytes

[0037] Rat cardiomyocytes H9C2 were evenly seeded into 12-well plates containing cell spreaders, with approximately 3.5 × 10⁻⁶ cells per well. 5 Cells adhered and grew for 24 h before being treated with PBS (pH=7.4), AngII (10 μM, 24 h), AngII (10 μM, pretreated for 2 h before adding Nodakenin) + Nodakenin (10 μM, 24 h) to obtain PBS treatment group, AngII treatment group, and AngII+NK treatment group.

[0038] Immunofluorescence staining experiments were performed on cardiac troponin (cTNT) separately. The results are as follows: Figure 1 As shown, from Figure 1 It can be seen that treatment of cardiomyocytes with 10 μM AngII significantly increased cell area, while the Nodakenin treatment group effectively reduced the AngII-induced increase in cell area. Therefore, direct in vitro treatment of cardiomyocytes with Nodakenin can significantly inhibit the AngII-induced increase in cardiomyocyte area, with no significant difference compared to the blank control group.

[0039] Example 2

[0040] Effects of Nodakenin direct in vitro treatment of cardiomyocytes on the expression of markers of myocardial hypertrophy in cells

[0041] Rat cardiomyocytes H9C2 were evenly seeded into 6-well plates, with approximately 1 × 10⁻⁶ cells per well. 6 After cells adhered and grew for 24 hours, they were treated with the drug as described in Example 1. After 24 hours of treatment in each group, total RNA was extracted from the cells, and the expression of mRNA markers of myocardial hypertrophy in cardiomyocytes was detected using quantitative real-time polymerase chain reaction (qRT-PCR).

[0042] from Figure 2 As can be seen, AngII treatment significantly increased the expression of myocardial hypertrophy markers, while direct treatment with 10 μM Nodakenin in vitro significantly and effectively reduced the expression of myocardial hypertrophy markers, indicating that Nodakenin has a significant anti-myocardial hypertrophy effect.

[0043] Example 3

[0044] Effects of Nodakenin gavage on heart and left ventricular weight in mice

[0045] In the progression of hypertrophic cardiomyopathy, pathological thickening of the heart and increased ventricular wall thickness lead to abnormal increases in both heart weight and left ventricular weight. This embodiment measures heart weight (WT) and Myh6 levels after oral administration of Nodakenin. R404Q The effects of Nodakenin on the progression of myocardial hypertrophy in vivo were examined by measuring the weight of the mouse heart and left ventricle.

[0046] Four-week-old wild-type mice (WT) and Myh6 gene R404Q point mutation-induced hypertrophic cardiomyopathy mice (Myh6) were used. R404Q A hypertrophic cardiomyopathy model was established using male R404Q mutant mice. After successful modeling, mice were randomly divided into a control group, a model group, a low-dose group (5 mg / kg) of imperatorin (20 mg / kg), a high-dose group (20 mg / kg), and a positive control group (metoprolol, 20 mg / kg). Mice were administered the drug once daily by gavage for 4 weeks, while maintaining a normal diet. The general condition of the mice was monitored during the administration process. Echocardiographic parameters and cardiac mass index (heart weight to tibia length ratio, HW / TL) were measured 24 hours after the last administration, and gross cardiac images were taken.

[0047] Figure 3 The results showed that, compared with the model group, the HW / TL and left ventricular weight were significantly reduced in the imperatorin treatment group, and from Figure 4 The heart images showed that the heart was significantly smaller after administration of Nodakenin, suggesting that it can significantly improve myocardial hypertrophy and alleviate dysfunction. Moreover, the high-dose purslane treatment group showed better gross improvement in the heart than the positive control group.

[0048] Example 4

[0049] Effects of Nodakenin gavage on heart and left ventricular thickness in mice

[0050] After feeding mice for one month using the method described in Example 3, echocardiography was performed on the mice, and the thickness of the left ventricular anterior wall during systole, diastole, systole, and diastole was calculated using the Vevo3100 system.

[0051] Test results as follows Figure 5 As shown (from left to right), from Figure 5The results showed that the left ventricular wall thickness in Myh6R404Q mice was significantly higher than that in WT mice during both diastole and systole, while Nodakenin administration significantly reduced the left ventricular wall thickness in Myh6R404Q mice. This indicates that Nodakenin has a beneficial effect in combating the progression of pathological myocardial hypertrophy in hereditary hypertrophic cardiomyopathy.

[0052] In summary, this invention, through in vivo and in vitro experiments, reveals for the first time that Nodakenin can significantly inhibit the AngII-induced cardiomyocyte hypertrophy phenotype, and in Myh6... R404Q Nodakenin significantly reduced heart weight-to-total ratio (HW / TL), left ventricular mass, and ventricular wall thickness in mice with hereditary hypertrophic cardiomyopathy, thereby alleviating pathological myocardial hypertrophy and improving cardiac structure and function. This invention is the first to reveal nodakenin as a candidate active ingredient for hereditary hypertrophic cardiomyopathy, demonstrating clear development value in terms of efficacy and safety of origin, and providing a novel drug option and technological pathway for the prevention and treatment of hypertrophic cardiomyopathy.

Claims

1. Application of purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy.

2. The application according to claim 1, characterized in that, The hypertrophic cardiomyopathy includes hereditary hypertrophic cardiomyopathy and the symptoms caused by hereditary hypertrophic cardiomyopathy.

3. The application according to claim 1, characterized in that, The application of the aforementioned purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by inhibiting the increase in cardiomyocyte area.

4. The application according to claim 1, characterized in that, The aforementioned purslane is used in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by reducing the expression of myocardial hypertrophy markers or by inhibiting the hypertrophic phenotype of cardiomyocytes.

5. The application according to claim 1, characterized in that, The application of the purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by reducing the weight of the heart or ventricles.

6. The application according to claim 1, characterized in that, The application of the purslane in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy by reducing ventricular wall thickness.

7. A pharmaceutical composition for treating hypertrophic cardiomyopathy, characterized in that, It includes purslane as the sole drug component for treating hypertrophic cardiomyopathy or in combination with other drug components for treating hypertrophic cardiomyopathy, as well as the raw materials, excipients or carriers required for the formulation.

8. The pharmaceutical composition for treating hypertrophic cardiomyopathy according to claim 7, characterized in that, The dosage form of the composition preferably includes emulsions, microemulsions, gels, solutions, tinctures, films, ointments, creams, or patches.

9. Use of the pharmaceutical composition of claim 7 in the preparation of a reagent or medicament for treating hypertrophic cardiomyopathy.

10. The use of the purslane of claim 1 or the composition of claim 7 in the preparation of a medicament for the prevention or treatment of symptoms caused by hypertrophic cardiomyopathy.

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