Use of AICAR in the preparation of drugs for preventing and / or treating uremic cardiomyopathy

Abstract

The application discloses application of AICAR in preparation of medicines for preventing and / or treating uremic cardiomyopathy, and belongs to the technical field of biological medicines. Through a 5 / 6 nephrectomy induced uremic cardiomyopathy mouse model and an indoxyl sulfate induced myocardial cell damage model, it is first confirmed that AICAR can significantly improve the heart function of uremic cardiomyopathy, improve the left ventricular ejection fraction and the short axis shortening rate, reduce the left ventricular posterior wall thickness, and reduce myocardial fibrosis and myocardial cell hypertrophy. Meanwhile, AICAR can repair the ultrastructure damage of mitochondria, restore the expression of oxidative phosphorylation related proteins, improve the mitochondrial respiration function, and systematically reverse the abnormal metabolism of six categories of lipids such as glycerolipids, glycerophospholipids and sphingolipids. The application provides a brand new drug intervention strategy for uremic cardiomyopathy, and has clear clinical conversion potential.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and cardiovascular disease prevention and treatment technology, specifically involving the application of AICAR (5-aminoimidazole-4-carboxamide-1-β-D-furanoside) in the preparation of drugs for the prevention and / or treatment of uremic cardiomyopathy, and is a pharmaceutical invention. Background Technology

[0002] Uremic cardiomyopathy is one of the most common and serious cardiovascular complications in patients with chronic kidney disease, especially end-stage renal disease. Epidemiological data show that more than 50% of patients with end-stage renal disease have varying degrees of myocardial structural and functional abnormalities, and cardiovascular events are the leading cause of death in dialysis patients, accounting for 40% to 50% of all deaths. Typical pathological manifestations of uremic cardiomyopathy include left ventricular hypertrophy, myocardial fibrosis, cardiomyocyte hypertrophy, decreased myocardial compliance, and systolic and / or diastolic dysfunction. Clinical outcomes include progressive heart failure and a significantly increased risk of sudden death.

[0003] Currently, clinical intervention strategies for uremic cardiomyopathy mainly revolve around controlling blood pressure, correcting volume overload, regulating mineral metabolism disorders, and renal replacement therapy (hemodialysis or kidney transplantation). While these measures can partially delay disease progression, they are all supportive treatments targeting upstream causes or systemic metabolic disorders, rather than directly targeting the pathological changes occurring in cardiomyocytes themselves in the uremic state. In other words, existing treatments lack specific reversal or repair effects on existing myocardial hypertrophy, fibrosis, and mitochondrial damage. To date, no drugs specifically targeting the core aspects of the pathogenesis of uremic cardiomyopathy—cardiomyocyte metabolic reprogramming and mitochondrial dysfunction—have been approved for marketing, indicating a clear unmet clinical need.

[0004] In recent years, an increasing number of studies have suggested that uremic cardiomyopathy is not simply caused by hemodynamic abnormalities or increased volume overload, but is closely related to intrinsic metabolic disorders in cardiomyocytes. In the microenvironment of long-term accumulation of uremic toxins (such as indophenol sulfate and p-cresol sulfate, which are protein-bound toxins), cardiomyocytes undergo significant metabolic reprogramming, manifested as impaired mitochondrial oxidative phosphorylation, inhibition of fatty acid β-oxidation pathways, abnormal lipid accumulation, and energy metabolism disorders. As the core organelle of cardiomyocyte energy metabolism, mitochondria exhibit ultrastructural damage under uremic stress, including swelling, cristae breakage, vacuolar degeneration, and disordered arrangement, further exacerbating insufficient myocardial energy supply and oxidative stress damage, forming a vicious cycle of "mitochondrial dysfunction—energy metabolism disorder—myocardial remodeling." Therefore, finding drugs that can directly target the restoration of mitochondrial function and correction of metabolic reprogramming in cardiomyocytes is of great significance for the specific treatment of uremic cardiomyopathy.

[0005] AICAR (5-aminoimidazole-4-carboxamide-1-β-D-furanoside, also known as acaldicin) is an adenosine analog that generates ZMP through intracellular phosphorylation, thereby activating AMP-activated protein kinase (AMPK) and participating in the regulation of cellular energy metabolism homeostasis. Previous studies have shown that AICAR exhibits certain cardioprotective effects in models of ischemic heart injury, diabetic cardiomyopathy, and heart failure induced by chemotherapy drugs (such as doxorubicin). However, the pathological mechanisms involved in these models are mainly related to acute ischemia-reperfusion injury, hyperglycemic toxicity, or direct cardiotoxicity of chemotherapy drugs, which are fundamentally different from chronic metabolic myocardial damage caused by long-term accumulation of uremic toxins. Currently, there are no literature reports or technological insights regarding whether AICAR can improve cardiac function impairment, myocardial remodeling, and mitochondrial dysfunction in uremic cardiomyopathy.

[0006] In view of this, this invention proposes for the first time that AICAR can be used to prepare drugs for the prevention and / or treatment of uremic cardiomyopathy, and verifies this through an animal model of uremic cardiomyopathy induced by 5 / 6 nephrectomy and a model of cardiomyocyte damage induced by uremic toxin, providing a new strategy and experimental basis for drug intervention in uremic cardiomyopathy. Summary of the Invention

[0007] In view of the shortcomings of existing interventions in addressing the lack of effective targeting of mitochondrial dysfunction and lipid metabolism reprogramming in uremic cardiomyopathy, and the fact that existing treatments are mostly limited to supportive measures such as controlling blood pressure and volume overload without directly repairing intrinsic damage to cardiomyocytes, this invention provides the application of AICAR in the preparation of drugs for the prevention and / or treatment of uremic cardiomyopathy.

[0008] To achieve the above objectives, the present invention provides the following technical solution: In a first aspect, the present invention provides the use of AICAR in the preparation of drugs for the prevention and / or treatment of uremic cardiomyopathy; wherein uremic cardiomyopathy is a myocardial structural and functional disorder secondary to chronic kidney disease, particularly end-stage renal disease.

[0009] Further, the AICAR is 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (CAS: 2627-69-2), also known as acadesine, or a pharmaceutically acceptable salt, solvate, prodrug, or pharmaceutically acceptable combination thereof. As an adenosine analog, AICAR can generate ZMP through intracellular phosphorylation, activating AMP-activated protein kinase, thereby participating in the regulation of cellular energy metabolism homeostasis.

[0010] Furthermore, the uremic cardiomyopathy is characterized by one or more of the following pathological changes: decreased cardiac function, left ventricular wall thickening, myocardial hypertrophy, myocardial fibrosis, mitochondrial dysfunction, and lipid metabolism disorders. Specifically, the decreased cardiac function is manifested as a decrease in left ventricular ejection fraction and / or a decrease in left ventricular shortening rate; the left ventricular wall thickening is manifested as an increase in the end-diastolic thickness of the left ventricular posterior wall and / or an increase in the end-systolic thickness of the left ventricular posterior wall; the mitochondrial dysfunction is manifested as mitochondrial swelling, vacuolar degeneration, cristae breakage, and / or disordered arrangement; and the lipid metabolism disorders are manifested as abnormal metabolism of glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl lipids, and / or isoprene lipids.

[0011] Furthermore, the AICAR is used to improve cardiac function and myocardial remodeling. Preferably, the improvement of cardiac function and myocardial remodeling includes increasing left ventricular ejection fraction and / or left ventricular shortening rate, reducing left ventricular posterior wall end-diastolic thickness and / or left ventricular posterior wall end-systolic thickness, and alleviating myocardial fibrosis and / or myocardial hypertrophy. In a 5 / 6 nephrectomy mouse model, AICAR (500 mg / kg, 3 times a week for 3 months) intervention significantly increased the left ventricular ejection fraction from 69.7±9.9% to 78.7±2.9% (P<0.05), the left ventricular fractional shortening from 38.9±7.5% to 46.4±2.7% (P<0.05), the myocardial fibrosis area from 12.2±4.8% to 7.7±3.2% (P<0.05), and the cardiomyocyte cross-sectional area from 384.0±57.5% to 308.6±39.8% (P<0.05).

[0012] Furthermore, the AICAR is used to improve mitochondrial structure and energy metabolism disorders. Preferably, the improvement of mitochondrial structure and energy metabolism disorders includes improving one or more mitochondrial ultrastructural abnormalities such as mitochondrial swelling, vacuolar degeneration, cristae breakage, or disordered arrangement, and increasing the expression levels of oxidative phosphorylation-related proteins. Transmission electron microscopy results showed that after AICAR intervention, the ultrastructural damage such as mitochondrial swelling, cristae breakage, and disordered arrangement in the myocardium of 5 / 6 nephrectomy model mice was significantly alleviated. Western blot analysis showed that AICAR can increase the protein expression levels of one or more oxidative phosphorylation complex subunits in ATP5A1, UQCRC, SDHB, and MTCO2.

[0013] Furthermore, the AICAR is used to improve myocardial lipid metabolism disorders. Preferably, improving myocardial lipid metabolism disorders includes reversing or partially correcting abnormal changes in one or more lipids among glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl lipids, and isoprene lipids. Lipomics results showed that 2874 metabolites belonging to 85 categories were detected in the myocardial tissue of 5 / 6 nephrectomy model mice, among which several lipid categories, including glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl lipids, and isoprene lipids, showed significant abnormalities. After AICAR intervention, the abnormal molecules in the above lipid categories showed varying degrees of reversal or regression, and principal component analysis (PCA) showed that the overall metabolite profile of the AICAR intervention group regressed towards that of the sham-operated group.

[0014] Furthermore, the AICAR is used to increase the expression levels of proteins related to fatty acid transport, the tricarboxylic acid cycle, and mitochondrial β-oxidation. Preferably, the AICAR increases the expression levels of one or more proteins selected from the following: CPT1, OGDH, PDK4, ACOX1, and ACOT1. In a 5 / 6 nephrectomy model, the above-mentioned proteins related to fatty acid uptake, transport, and β-oxidation were downregulated in the model group, but their expression levels increased after AICAR intervention.

[0015] Furthermore, the AICAR is administered via intraperitoneal injection at a dose of 500 mg / kg body weight, three times a week for a period of three months. In a 5 / 6 nephrectomy mouse model, AICAR was administered starting one week post-surgery and continued until 12 weeks post-surgery. Significant improvements were observed in all the aforementioned indicators of cardiac function, pathology, mitochondrial structure, and lipid metabolism. This dosing regimen has been validated in vivo in demonstrating that AICAR effectively improves uremic cardiomyopathy, and the animals tolerated it well with no obvious toxic reactions observed.

[0016] Furthermore, the AICAR is used to improve uremic toxin-induced mitochondrial respiratory dysfunction in cardiomyocytes. Specifically, the uremic toxin includes indophenol sulfate. In a neonatal rat ventricular myocyte (NRVM) model induced by indophenol sulfate (1 mM), after 24 hours of AICAR (1 mM) intervention, the Seahorse mitochondrial stress test showed that basal respiration, ATP-related respiration, and maximal respiration were significantly reduced in the indophenol sulfate group, while the above mitochondrial respiratory function indicators were significantly restored in the AICAR combined treatment group, suggesting that AICAR can directly improve the damage to cardiomyocyte mitochondrial respiratory function caused by uremic toxins.

[0017] Furthermore, the drug uses AICAR as its main active ingredient, and also contains one or more of a pharmaceutically acceptable carrier, diluent, buffer, stabilizer, or excipient. The drug is available in the form of an injection, a lyophilized powder for injection, or an oral formulation.

[0018] In a second aspect, the present invention also provides a pharmaceutical composition for the prevention and / or treatment of uremic cardiomyopathy, comprising an effective dose of AICAR and a pharmaceutically acceptable carrier, said pharmaceutical composition being an injection, a lyophilized powder injection, or an oral formulation.

[0019] Compared with the prior art, the present invention has the following outstanding advantages: (1) This invention first proposes and verifies a novel use of AICAR in the preparation of drugs for the prevention and / or treatment of uremic cardiomyopathy. While existing technologies have reported cardioprotective effects of AICAR in models of ischemic heart injury, diabetic cardiomyopathy, and doxorubicin-induced heart failure, the pathological mechanisms of these models are mainly related to acute ischemia-reperfusion injury, hyperglycemic toxicity, or direct myocardial toxicity from chemotherapy drugs, which are fundamentally different from chronic metabolic myocardial injury caused by long-term accumulation of uremic toxins. Currently, there are no reports of AICAR being used for uremic cardiomyopathy; this invention fills this technological gap.

[0020] (2) This invention has verified the protective effect of AICAR in both in vivo animal models and in vitro cell models. In a mouse model of uremic cardiomyopathy induced by 5 / 6 nephrectomy, AICAR significantly improved cardiac function, reduced myocardial hypertrophy and fibrosis, and repaired mitochondrial ultrastructural damage; in a cardiomyocyte injury model induced by indophenol sulfate, AICAR significantly improved mitochondrial respiratory dysfunction. These results provide sufficient experimental evidence for the treatment of uremic cardiomyopathy with AICAR.

[0021] (3) This invention reveals for the first time the systemic corrective effect of AICAR on lipid metabolism disorders in uremic cardiomyopathy. Through lipidomics technology, this invention found that AICAR can restore the overall lipid metabolism profile of uremic cardiomyopathy to a normal state, encompassing a broad reversal of six major lipid classes, including glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl lipids, and isoprene lipids. This systemic remodeling of the entire lipid metabolism network in myocardial tissue provides a novel mechanistic insight for AICAR treatment of uremic cardiomyopathy.

[0022] (4) This invention further reveals the mechanism of action of AICAR: In the uremic environment, AICAR repairs the function of the mitochondrial electron transport chain by restoring the expression of oxidative phosphorylation complex (ATP5A1, UQCRC, SDHB, MTCO2) proteins, while promoting lipid utilization by upregulating the expression of fatty acid transport and β-oxidation-related proteins (CPT1, OGDH, PDK4, ACOX1, ACOT1), thus achieving a dual improvement in mitochondrial function and lipid metabolism. This multi-target action characteristic makes it superior to single-target intervention strategies.

[0023] (5) The dosing regimen provided by this invention (500 mg / kg, 3 times a week for 3 months) has been experimentally verified to be safe and effective, with good animal tolerance and clear clinical translation potential. As a drug that has entered Phase III clinical trials, AICAR has sufficient human data to support its pharmacokinetics and safety, which can significantly shorten the research and development cycle and reduce the risk of translation.

[0024] In summary, this invention provides a drug intervention program for uremic cardiomyopathy centered on AICAR, which has a clear mechanism of action, multi-level experimental data support, and good prospects for clinical translation, providing a novel drug strategy and experimental basis for the treatment of uremic cardiomyopathy. Attached Figure Description

[0025] Figure 1 M-mode echocardiography and statistical graphs of cardiac function and structural parameters are shown for each group of mice. A represents representative M-mode echocardiograms of each group; B shows the measurement results of left ventricular ejection fraction (EF%); C shows the measurement results of left ventricular fractional shortening (FS%); D shows the measurement results of left ventricular posterior wall end-diastolic thickness (LVPW d); and E shows the measurement results of left ventricular posterior wall end-systolic thickness (LVPW s). The horizontal axis represents the experimental group, and the vertical axis represents the corresponding indicator values, marked with statistical significance markers. Figure 2 The images show the histopathological staining and quantitative analysis of myocardial tissue from each group of mice. A represents the representative histological staining and ultrastructural images of myocardial tissue from each group of mice, from top to bottom: HE staining of myocardial cross-section (scale bar 1.25 mm), HE high-power field (scale bar 100 μm), Masson's trichrome staining (scale bar 50 μm), WGA fluorescence staining (scale bar 50 μm), and transmission electron microscopy (TEM, scale bar 500 nm); B is a quantitative statistical chart of the percentage of myocardial fibrosis area based on Masson's staining; C is a quantitative statistical chart of the cross-sectional area (CSV) of cardiomyocytes based on WGA staining. The horizontal axis represents the experimental group, and the vertical axis represents the quantitative value of the corresponding indicator. Figure 3Images show the ultrastructure of myocardial mitochondria and the expression of oxidative phosphorylation-related proteins in each group of mice. A shows the ultrastructural images of myocardial mitochondria in each group of mice under transmission electron microscopy (TEM); B shows representative bands of oxidative phosphorylation (OXPHOS) complex-related proteins (ATP5A1, UQCRC, SDHB, MTCO2, NDUFB8) and the internal control GAPDH in Western blotting; C shows the quantitative statistical graph of the relative expression levels of the above oxidative phosphorylation-related proteins. The horizontal axis represents the experimental group, and the vertical axis represents the relative protein expression level. Figure 4 This figure shows the mitochondrial respiratory function of neonatal rat ventricular myocytes (NRVMs) under different treatment conditions. A shows the dynamic change curve of real-time oxygen consumption rate (OCR) during the Seahorse mitochondrial stress test, with the addition time points of oligomycin, FCCP, and rotenone & antimycin A indicated. B is the OCR quantitative statistical graph of basal respiration. C is the OCR quantitative statistical graph of ATP production-related respiration. D is the OCR quantitative statistical graph of maximum respiration. The horizontal axis represents the experimental group, and the vertical axis represents the oxygen consumption rate. The legend indicates the control group, AICAR group, indophenol sulfate (IS) group, and AICAR+IS combined treatment group. Figure 5 Figures show the results of lipidomics analysis of myocardial tissue from each group of mice. A is the principal component analysis (PCA) score of myocardial lipidomics; B is the circular distribution statistical diagram of the 2874 detected metabolites and their corresponding 85 lipid superclasses; C is the heatmap of differential metabolites in six major lipid classes (glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl lipids, and isoprene lipids) and the Log2FC fold change plot comparing the AICAR intervention group and the model group. The horizontal axis in the figures indicates different groups and different lipid molecules. Figure 6 The figures show the expression results of lipid metabolism-related proteins and genes in the myocardial tissue of mice in each group. A represents the representative bands of fatty acid transport-related protein CPT1, tricarboxylic acid cycle rate-limiting enzyme OGDH, and internal control GAPDH in Western blotting; B is a quantitative statistical graph of the relative expression levels of CPT1 and OGDH proteins; C is a quantitative statistical graph of the relative expression levels of mRNAs related to fatty acid uptake and transport (CPT1, PDK4) and mitochondrial β-oxidation (ACOX1, ACOT1). The horizontal axis represents experimental groups, and the vertical axis represents the relative expression levels of proteins or mRNAs. Detailed Implementation

[0026] The present invention will be further described below with reference to specific embodiments and accompanying drawings, but the scope of protection of the present invention is not limited to the following embodiments. Without departing from the concept of the present invention, those skilled in the art can make conventional substitutions or adjustments to specific experimental conditions, routes of administration, and detection methods, all of which should fall within the scope of protection of the present invention.

[0027] Animal experiments were approved by the Experimental Animal Management and Use Committee of Nanjing Medical University (ethics approval number: IACUC-2302033).

[0028] Example 1 5 / 6 nephrectomy model and AICAR administration.

[0029] A 5 / 6 nephrectomy (5 / 6Nx) animal model was established, with four subgroups: Vehicle + Sham, AICAR + Sham, Vehicle + 5 / 6Nx, and AICAR + 5 / 6Nx. Male wild-type C57BL / 6J mice were purchased from Jicui Pharmaceutical Co., Ltd. (Nanjing, China), acclimatized for one week before the experiment, and were 7 weeks old at the time of the experiment. AICAR (Sigma-Aldrich, A9978) was dissolved in DMSO to prepare a 20 mM stock solution and stored at −80 °C. In the 5 / 6Nx model, mice were intraperitoneally injected three times a week with AICAR at a dose of 500 mg / kg, starting one week after nephrectomy, for three months.

[0030] After the drug administration cycle, echocardiography, histological staining, mitochondrial ultrastructural observation, protein expression detection, and lipidomics analysis were performed on animals in each group. The results showed that compared with the 5 / 6Nx group, the AICAR intervention group showed an improvement trend in cardiac function, myocardial remodeling, mitochondrial damage, and lipid metabolism.

[0031] Example 2 AICAR improves cardiac function and myocardial remodeling.

[0032] Echocardiography was used to evaluate cardiac function and left ventricular structural changes in each group of animals.

[0033] In the 5 / 6Nx mouse model, AICAR significantly reversed myocardial injury. Compared with the Vehicle+Sham group, echocardiography showed that the left ventricular ejection fraction (EF) was significantly reduced in the Vehicle+5 / 6Nx group (69.7 ± 9.9% vs 80.9 ± 2.4%, P<0.01), and the left ventricular fractional shortening (FS) was also significantly reduced (38.9 ± 7.5% vs 48.8 ± 2.5%, P<0.01). Compared with the Vehicle+5 / 6Nx group, echocardiography showed that the left ventricular ejection fraction (EF) was significantly increased to 78.7 ± 2.9% (P<0.05), and the left ventricular fractional shortening (FS) was also significantly increased to 46.4 ± 2.7% (P<0.05). The results showed that the 5 / 6Nx group exhibited decreased EF and FS, and increased LVPWd and LVPWs; after AICAR intervention, these abnormal indicators were significantly improved (e.g., Figure 1 (As shown in AE).

[0034] Further myocardial tissue staining was used to evaluate the degree of myocardial hypertrophy and fibrosis. Compared with the Vehicle+Sham group, the Vehicle+5 / 6Nx group showed disordered arrangement of cardiomyocytes, increased fibrosis area (12.2 ± 4.8% vs 3.0 ± 1.8%, P<0.05), and increased cardiomyocyte cross-sectional area (384.0 ± 57.5% vs 208.3 ± 37.9%, P<0.05). In contrast, compared with the Vehicle+5 / 6Nx group, the AICAR+5 / 6Nx group showed a significant decrease in fibrosis area to 7.7 ± 3.2% (P<0.05) and a significant decrease in cardiomyocyte cross-sectional area to 308.6 ± 39.8% (P<0.05). The results indicate that AICAR intervention significantly reduced the degree of myocardial hypertrophy and fibrosis (e.g., ...). Figure 2 (As shown in AC). The above results support the use of AICAR in improving cardiac function and myocardial remodeling.

[0035] Example 3 AICAR improves mitochondrial structure and energy metabolism abnormalities.

[0036] Transmission electron microscopy was used to observe the ultrastructure of myocardial mitochondria in each group of animals. The results showed that in the 5 / 6Nx group, mitochondria exhibited significant swelling, vacuolar degeneration, fragmentation, and disordered arrangement. After AICAR intervention, these abnormalities were significantly alleviated, suggesting that AICAR can effectively improve mitochondrial structural damage associated with uremic cardiomyopathy (e.g., mitochondrial structure). Figure 3 (As shown in A).

[0037] Further analysis was conducted to examine the expression levels of oxidative phosphorylation and metabolism-related proteins. The results showed that AICAR significantly increased the expression levels of one or more proteins among ATP5A1, UQCRC, SDHB, and MTCO2 (e.g., ...). Figure 3 As shown in B and 3C), it can also effectively upregulate the expression of one or more proteins among CPT1, OGDH, PDK4, ACOX1, and ACOT1 (e.g., Figure 6 As shown in A and 6B, AICAR can effectively improve mitochondrial energy metabolism disorders and promote fatty acid transport and oxidative metabolism.

[0038] Example 4 AICAR improves uremic toxin-induced energy metabolism disorders or mitochondrial respiratory dysfunction in neonatal rat cardiomyocytes.

[0039] Neonatal rat ventricular myocytes (NRVMs) were isolated from the ventricles of 1- to 3-day-old Sprague-Dawley rats using a sequential enzymatic digestion method involving trypsin and collagenase II. The discretized cells were pre-coated for 1-2 hours to reduce fibroblast contamination through differential adhesion. Subsequently, non-adherent cardiomyocytes were seeded onto coated culture plates or coverslips and maintained in a humid environment at 37°C with 5% CO2 in DMEM / F12 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. After 24 hours, the medium was replaced with maintenance medium, and the cells were treated according to the experimental design.

[0040] The oxygen consumption rate (OCR) of neonatal rat ventricular myocytes (NRVMs) under different treatment conditions was measured using the Seahorse mitochondrial stress test to evaluate mitochondrial respiration and energy metabolism. The experiment was divided into a control group, an AICAR group, an indophenol sulfate group (IS group), and an AICAR + IS group. Patients were treated with DMSO, 1 mM AICAR (Sigma-Aldrich, A9978), 1 mM IS (Sigma-Aldrich, I3875), or a combination of both for 24 h. Mitochondrial stress was measured using a Seahorse cellular energy metabolism analyzer. Oligomycin, FCCP, and rotenone / antimycin A were added sequentially, and the OCR of each group was measured. Basal respiration, ATP-related respiration, and maximal respiration (e.g., ) were calculated. Figure 4 (As shown in AD). The results showed that IS significantly reduced basal respiration, ATP-related respiration, and maximal respiration in NRVMs, suggesting that IS can induce mitochondrial respiration and energy metabolism disorders in cardiomyocytes; combined treatment with AICAR and IS significantly improved all OCR indicators compared to the IS group. The results indicate that AICAR can effectively improve IS-induced mitochondrial respiratory dysfunction in cardiomyocytes.

[0041] Example 5 AICAR improves myocardial lipid metabolism reprogramming.

[0042] Lipomics analysis was performed on myocardial tissues from the Sham group, the 5 / 6Nx group, and the 5 / 6Nx+AICAR group (e.g., Figure 5 (As shown in AC). Principal component analysis showed a significant separation between the 5 / 6Nx group and the Sham group, while the overall metabolomic profile of the AICAR intervention group regressed towards the Sham group ( Figure 5 A) suggests that AICAR can effectively partially correct myocardial lipid metabolism disorders caused by 5 / 6 nephrectomy.

[0043] Further classification and statistical results showed that a total of 2,874 metabolites were detected, belonging to 85 superclasses ( Figure 5 B). Differential analysis showed that the 5 / 6Nx group mainly involved abnormal changes in multiple lipid categories, including glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl groups, and isoprene lipids; after AICAR intervention, the abnormal molecules in the above lipid categories showed varying degrees of reversal or regression. Figure 5 C). These results support the use of AICAR in significantly improving lipid metabolism reprogramming.

[0044] Example 6 AICAR enhances fatty acid transport and the expression of β-oxidation-related proteins.

[0045] Western blotting and quantitative analysis were used to evaluate the expression of proteins and genes related to fatty acid transport, the tricarboxylic acid cycle, and mitochondrial β-oxidation (e.g., Figure 6 (As shown in AC). The results are as follows: Figure 6 As shown, the expression of CPT1 and OGDH proteins decreased in the 5 / 6Nx group, and significantly recovered after AICAR intervention. Figure 6 A, 6B). Meanwhile, proteins and genes related to fatty acid uptake, transport, and β-oxidation, such as CPT1, PDK4, ACOX1, and ACOT1, were downregulated in the 5 / 6Nx group, but their expression levels significantly increased after AICAR intervention. Figure 6 C). The above results further demonstrate that AICAR can significantly improve energy metabolism disorders and lipid metabolism disturbances associated with uremic cardiomyopathy.

[0046] In summary, AICAR can simultaneously improve cardiac function, alleviate myocardial remodeling, repair mitochondrial structural damage, enhance energy metabolism, and correct lipid metabolism disorders in uremic cardiomyopathy.

[0047] Therefore, AICARs can be used to prepare drugs for the prevention and / or treatment of uremic cardiomyopathy, with clear experimental support and promising application prospects.

[0048] 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 above embodiments do not limit the present invention in any way, and all technical solutions obtained by equivalent substitution or equivalent transformation fall within the protection scope of the present invention.

Claims

1. Application of AICAR in the preparation of drugs for the prevention and / or treatment of uremic cardiomyopathy.

2. The application according to claim 1, characterized in that, The AICAR is 5-aminoimidazol-4-carboxamide-1-β-D-furanoside, or a pharmaceutically acceptable salt, solvate, prodrug, or pharmaceutically acceptable composition thereof.

3. The application according to claim 1, characterized in that, Uremic cardiomyopathy is characterized by one or more of the following pathological changes: decreased cardiac function, thickening of the left ventricular wall, myocardial hypertrophy, myocardial fibrosis, mitochondrial dysfunction, and lipid metabolism disorder.

4. The application according to claim 1, characterized in that, The drug is used to improve cardiac function and myocardial remodeling, including increasing left ventricular ejection fraction and / or left ventricular shortening rate, reducing left ventricular posterior wall end-diastolic thickness and / or left ventricular posterior wall end-systolic thickness, and alleviating myocardial fibrosis and / or myocardial hypertrophy.

5. The application according to claim 1, characterized in that, The drug is used to improve mitochondrial structural and energy metabolism disorders, including improving one or more mitochondrial ultrastructural abnormalities such as mitochondrial swelling, vacuolar degeneration, cristae breakage, or disordered arrangement.

6. The application according to claim 1, characterized in that, The drug is used to improve myocardial lipid metabolism disorders, including reversing or partially correcting abnormal changes in one or more lipids, such as glycerides, glycerophospholipids, sphingolipids, sterol esters, fatty acyl esters, and isoprene esters.

7. The application according to claim 1, characterized in that, The drug increases the expression level of one or more proteins selected from the following: ATP5A1, UQCRC, SDHB, MTCO2, CPT1, OGDH, PDK4, ACOX1, and ACOT1.

8. The application according to claim 1, characterized in that, The dosage of AICAR is 500 mg / kg based on body weight, and the frequency of administration is 3 times per week.

9. A pharmaceutical composition for the prevention and / or treatment of uremic cardiomyopathy, characterized in that, It includes AICAR and pharmaceutically acceptable carriers.

10. The pharmaceutical composition according to claim 9, characterized in that, The dosage form of the pharmaceutical composition is an injection, a lyophilized powder for injection, or an oral preparation; the pharmaceutically acceptable carrier includes one or more of a diluent, a buffer, a stabilizer, or an excipient.

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