A pharmaceutical composition for alleviating stress load-induced heart disease

Abstract

This invention belongs to the field of biomedical technology and relates to a pharmaceutical composition for alleviating stress-induced cardiac disease. The invention provides a pharmaceutical composition for alleviating stress-induced cardiac disease, comprising oxalic acid, coenzyme Q, and an NLRP3 inhibitor. This invention verifies that the pharmaceutical composition can significantly inhibit human valvular interstitial cell calcification and exhibits excellent synergistic therapeutic effects compared to a single NLRP3 inhibitor.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a pharmaceutical composition for relieving stress-induced heart disease. Background Technology

[0002] Aortic valve calcification is a common type of heart disease and a significant cause of cardiovascular disease and death in an aging society. Prolonged abnormal pressure load on the heart triggers abnormal osteogenic differentiation and calcium deposition in valvular interstitial cells. This is the core pathological mechanism in the development of calcific valvular heart disease. During this process, the NLRP3 inflammasome is significantly activated, further mediating a vicious cycle between inflammation and calcification, ultimately leading to valvular sclerosis and stenosis, resulting in myocardial hypertrophy, heart failure, and even sudden death.

[0003] Current clinical treatments for pressure-overload-induced calcified valvular heart disease, especially aortic valve calcification, have significant limitations: traditional drugs mainly focus on symptomatic control of cardiac function and relief of hemodynamic abnormalities, lacking specific drugs that target and inhibit valvular interstitial cell calcification; although studies have shown that NLRP3 inhibitors (such as MCC950) can delay calcification by inhibiting inflammasome activation, their onset of action is slow and their calcification inhibition effect is limited when used alone, making it difficult to meet clinical treatment needs; oxalic acid has antioxidant, anti-inflammatory, and cell differentiation-inhibiting effects, while coenzyme Q can improve myocardial energy metabolism and assist in anti-inflammatory treatment. Although studies on these two alone or in combination have been reported, no studies have combined oxalic acid, coenzyme Q, and NLRP3 inhibitors, nor have they confirmed the therapeutic effect of this combination in pressure-overload-induced valvular calcification, and the synergistic calcification inhibition ratio has not been clarified.

[0004] In summary, the existing technology lacks a drug composition that can efficiently and rapidly target and inhibit human valvular interstitial cell calcification and has a synergistic therapeutic effect on pressure-overload-induced calcified valvular heart disease. Developing a compound drug with a clear target, scientific compatibility, and significant effect has become a key need to solve the clinical treatment challenges of this type of heart disease. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, this invention provides a pharmaceutical composition for alleviating stress-induced cardiac disease. This invention verifies that this pharmaceutical composition can significantly inhibit human valvular interstitial cell calcification, exhibiting excellent synergistic therapeutic effects compared to single NLRP3 inhibitors.

[0006] On one hand, the present invention relates to a pharmaceutical composition for relieving stress-induced heart disease, which consists of oxalic acid, coenzyme Q and NLRP3 inhibitor.

[0007] Optionally, in the pharmaceutical composition for relieving stress-induced heart disease provided by the present invention, the NLRP3 inhibitor is MCC950.

[0008] Furthermore, in the pharmaceutical composition for relieving stress-induced heart disease provided by the present invention, the molar ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 1~10:10~30:1~10.

[0009] Furthermore, in the pharmaceutical composition for relieving stress-induced heart disease provided by the present invention, the molar ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 5~10:20~30:5~10.

[0010] Furthermore, in the pharmaceutical composition for relieving stress-induced heart disease provided by the present invention, the molar ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 5:20:5.

[0011] On the other hand, the present invention relates to the use of the pharmaceutical composition for relieving stress-induced heart disease in the preparation of a medicament for the prevention and treatment of heart disease.

[0012] Furthermore, in the application provided by this invention, the heart disease is calcific valvular heart disease.

[0013] Furthermore, in the application provided by this invention, the heart disease is aortic valve calcification.

[0014] Furthermore, in the application provided by the present invention, the concentration ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor, in μM, is 1~10:10~30:1~10.

[0015] Furthermore, in the application provided by the present invention, the concentration ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor, in μM, is 5~10:20~30:5~10.

[0016] Furthermore, in the application provided by the present invention, the concentration ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 5:20:5 in μM.

[0017] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0018] The pharmaceutical composition of this invention overcomes the therapeutic limitations of any combination of a single NLRP3 inhibitor, oxalic acid, and coenzyme Q / NLRP3 inhibitor / coenzyme Q. It can more rapidly and efficiently inhibit abnormal osteogenic differentiation of human valvular interstitial cells, significantly reduce the expression of osteogenic-related genes ALP and RUNX2, and effectively reverse the process of extrinsic-induced calcium deposition. In contrast, the combination of individual components only shows a slight gene expression inhibition effect after 7 days of action, and oxalic acid has no significant therapeutic effect when replaced by oxalic acid. This demonstrates that the combination of oxalic acid, coenzyme Q, and NLRP3 inhibitor is the core strategy for achieving calcification inhibition, and its synergistic effect is significantly better than existing single or combination methods. The composition contains oxalic acid, a natural bioactive substance; coenzyme Q, an energy metabolism substance naturally present in the human body and a commonly used excipient in clinical practice; and NLRP3 inhibitors (such as MCC950), which are mature targeted anti-inflammatory inhibitors. The combination of the three components has no antagonistic effect and the safety is controllable. Furthermore, all components are commercially available raw materials, making production costs controllable and facilitating subsequent preparation into various dosage forms such as oral and injectable formulations. It has good prospects for industrial development and clinical application. Attached Figure Description

[0019] To more clearly illustrate the technical solution of the present invention, the accompanying drawings involved in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the description are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a graph showing the improvement results of the drug combination in Example 1.

[0021] Figure 2 This is a graph showing the improvement results of the drug combination in Example 2. Detailed Implementation

[0022] The technical solution of the present invention will be described below with reference to embodiments. However, the present invention is not limited to the following embodiments. Unless otherwise specified, the experimental and detection methods described in each embodiment are conventional methods; the reagents and materials described are commercially available unless otherwise specified. Unless otherwise specified, all percentages in the following embodiments refer to mass percentages. Unless otherwise specified, all proportions in the following embodiments refer to mass ratios.

[0023] Example 1 This embodiment provides experimental results on the use of a pharmaceutical composition to inhibit calcification of human valvular interstitial cells.

[0024] (1) Constructing a human aortic valve interstitial cell calcification model Human aortic valve interstitial cells were induced to calcify using a calcification medium containing 50 µg / mL ascorbic acid and 2.5 mM inorganic phosphate. This medium was prepared from NaH₂PO₄ / Na₂HPO₄ (pH=7.4). The control group used an equal volume of phosphate buffer.

[0025] (2) Experimental grouping and experimental methods Based on the human aortic valve interstitial cell calcification model constructed in (1), it was divided into 5 groups: blank control group (normal culture medium), model group (induced calcification), low-dose group (induced calcification + 1 μM oxalate + 10 μM coenzyme Q + 1 μM MCC950), medium-dose group (induced calcification + 5 μM oxalate + 20 μM coenzyme Q + 5 μM MCC950), and high-dose group (induced calcification + 10 μM oxalate + 30 μM coenzyme Q + 10 μM MCC950). The relative expression changes of osteogenic related genes ALP and RUNX2 were examined by RT-PCR after 3 days and 7 days of treatment, respectively, with GAPDH gene as internal reference gene. Each group was repeated 5 times (n=5). The concentrations in each group are expressed as the final concentration in the culture medium.

[0026] (3) Experimental results Depend on Figure 1 The experimental results showed that, compared with the blank control group, the relative expression levels of ALP and RUNX2 genes in the model group were significantly increased at 3 and 7 days, and continued to increase over time. The low-dose, medium-dose, and high-dose groups all restored ALP and RUNX2 gene expression levels to near those of the blank control group, with the medium-dose and high-dose groups being even closer to the blank control group. This indicates that the combination of oxalic acid, coenzyme Q, and NLRP3 inhibitor provided in this invention can improve exogenous induced calcification, reverse abnormal bone differentiation and calcium deposition, and is suitable for the prevention and treatment of aortic valve calcification.

[0027] Example 2 This example provides the effects of other drug combinations.

[0028] The experimental procedure was the same as in Example 1, with control group 1# (calcification induced + 30 μM coenzyme Q + 10 μM MCC950), control group 2# (calcification induced + 10 μM sarcosinate + 10 μM MCC950), control group 3# (calcification induced + 10 μM sarcosinate + 30 μM coenzyme Q), and control group 4# (calcification induced + 10 μM sarcosinate + 30 μM coenzyme Q + 10 μM MCC950).

[0029] Depend on Figure 2The experimental results showed that, although control groups 1#~4# reduced the expression of ALP and RUNX2 genes at 3 days, the reduction was not significant. At 7 days, control groups 1#~4# significantly reduced the expression of ALP and RUNX2 genes, but the effect was still lower than that of the combination of oxalic acid, coenzyme Q, and NLRP3 inhibitor. There was no significant difference between control groups 1# and control group 4#.

[0030] The above experimental results show that, compared with the use of coenzyme Q and MCC950 alone, oxalic acid and MCC950 alone, or oxalic acid and coenzyme Q alone, the combination of oxalic acid, coenzyme Q, and NLRP3 inhibitor provided by this invention can more rapidly and effectively inhibit osteogenic differentiation of human valvular interstitial cells and reduce the expression of ALP and RUNX2 genes, and the effect is significantly better than the combination of two at the same concentration. Furthermore, the effect of oxalic acid replaced by oxalool is comparable to that without oxalic acid, indicating that the combination of the three is the optimal approach. Therefore, the pharmaceutical composition of this invention has a synergistic effect in the prevention and treatment of aortic valve calcification.

[0031] The embodiments described above are only some, not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate preferred embodiments. All other embodiments obtained by those skilled in the art through related deductions and substitutions based on the inventive concept, without inventive effort, are within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for relieving stress-induced heart disease, characterized in that, It consists of oxalic acid, coenzyme Q and NLRP3 inhibitor.

2. The pharmaceutical composition for alleviating stress load-induced heart disease according to claim 1, wherein The molar ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 1~10:10~30:1~10.

3. The pharmaceutical composition for alleviating stress load-induced heart disease according to claim 2, wherein The molar ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 5~10:20~30:5~10.

4. The pharmaceutical composition for alleviating stress load-induced heart disease according to claim 3, wherein The molar ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor is 5:20:

5.

5. The use of the pharmaceutical composition for relieving stress-induced heart disease according to any one of claims 1 to 4 in the preparation of a medicament for the prevention and treatment of heart disease.

6. Use according to claim 5, characterized in that, The heart disease in question is calcified valvular heart disease.

7. The application according to claim 6, characterized in that, The aforementioned heart disease is aortic valve calcification.

8. The application according to claim 7, characterized in that, The concentration ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor in μM was 1~10:10~30:1~10.

9. The application according to claim 8, characterized in that, The concentration ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor in μM was 5~10:20~30:5~10.

10. The application according to claim 9, characterized in that, The concentration ratio of oxalic acid, coenzyme Q and NLRP3 inhibitor in μM was 5:20:5.

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