Use of di'aoxinxuankang in the preparation of a drug for preventing adriamycin-induced cardiac endothelial injury toxicity

Abstract

The application provides a use of Di'ao Xin-xue-kang in preparation of a medicine for preventing doxorubicin-induced cardiac endothelial injury toxicity, and belongs to the field of medicines. The application uses Di'ao Xin-xue-kang for preventive administration, protects early cardiac endothelial injury induced by doxorubicin, and further can rescue or avoid occurrence of subsequent irreversible cardiotoxicity, especially uses Di'ao Xin-xue-kang for prevention or / and intervention of asymptomatic cardiac dysfunction related to cancer treatment, and opens a new use and administration method for clinical use of Di'ao Xin-xue-kang.

Description

Technical Field

[0001] This invention relates to the use of Di'ao Xinxuekang in the preparation of a drug for preventing doxorubicin-induced cardiac endothelial injury toxicity, and belongs to the pharmaceutical field. Background Technology

[0002] Doxorubicin (DOX) is an anthracycline chemotherapy drug effective in treating various malignant tumors, but its clinical application is limited by its dose-dependent cardiotoxicity. The *Guidelines for Cardio-Oncology* list the cardiotoxicity caused by this drug as the leading cause of treatment-related cardiovascular toxicity (CTR-CVT) among more than ten anti-tumor therapies, highlighting the significant clinical importance of early monitoring and proactive prevention of its cardiotoxicity. Doxorubicin cardiotoxicity is a complex pathological process involving multiple molecules and factors. Traditional views attribute it to oxidative stress and lipid, DNA, and protein damage caused by topoisomerase IIβ. Extensive research has been conducted on mechanisms such as mitochondrial damage, autophagy, apoptosis, and ferroptosis in cardiomyocytes, but these have not yet translated into effective clinical prevention and treatment methods. Previous studies have primarily focused on its direct effects on cardiomyocytes, which cannot fully explain the complex mechanisms of DOX cardiotoxicity. This is because the heart is a multicellular organ composed of interacting cardiomyocytes and non-cardiomyocytes, such as endothelial cells and fibroblasts. Although cardiomyocytes constitute the majority of the heart, making up 75% of its total volume, they only account for 30% of the total number of cells. The remaining 70% are non-cardiomyocytes, which are closely related to cardiomyocytes in both structure and function. Increasing research is focusing on non-cardiomyocytes, revealing that crosstalk between cardiomyocytes and non-cardiomyocytes is crucial in DOX-induced cardiotoxicity, with cardiac vascular endothelial cells being a particular focus of study. Recent studies suggest that DOX induces excessive ROS production that attacks cardiomyocytes while also directly affecting cardiac vascular endothelial cells and disrupting their crosstalk with cardiomyocytes.

[0003] Di'ao Xinxuekang Capsules (abbreviated as Xinxuekang or DXXK) are a new traditional Chinese medicine drug approved in 1988, consisting of the effective part of the plant Dioscorea opposita (Dioscorea opposita). Dioscorea nipponicaComposed of rhizome extract of *Makino*, mainly containing steroidal saponins such as pseudoprodiosgenin, it can promote blood circulation, remove blood stasis, regulate qi, and relieve pain, and is used for coronary heart disease with blood stasis syndrome. Recent clinical and pharmacological studies have shown that it has good efficacy in treating atherosclerosis and myocardial ischemia-reperfusion injury. For example, the literature includes: Chen Hong, et al., Study on the protective effect and mechanism of Di'ao Xinxuekang on myocardial ischemia-reperfusion injury, Chinese Traditional and Herbal Drugs, Vol. 41, No. 12, December 2010, which shows that the protective effect of Di'ao Xinxuekang on myocardial ischemia-reperfusion injury in rats is related to its antioxidant damage, inhibition of cardiomyocyte apoptosis and improvement of vascular endothelial function; Hu Yuqian, et al., Protective effect of Di'ao Xinxuekang-containing serum on H2O2-induced apoptosis of human umbilical vein endothelial cells, Chinese Traditional and Herbal Drugs Pharmacology and Clinical Practice, 2013;29(4), which shows that Di'ao Xinxuekang can protect vascular endothelial cells by increasing the survival rate of human umbilical vein endothelial cells treated with H2O2, reducing Caspase-3 activity and inhibiting the cell apoptosis pathway; Deng Liangyan, et al., Study on the protective effect of Di'ao Xinxuekang on doxorubicin-induced cardiotoxicity in zebrafish, Chinese Traditional and Herbal Drugs and Clinical Practice, 2025;16(2), which shows that Di'ao Xinxuekang has a certain protective effect on DOX-induced cardiotoxicity in zebrafish.

[0004] Among them, Deng Liangyan's published literature directly addresses the doxorubicin cardiotoxicity model in CTR-CVT (tumor therapy-related cardiovascular toxicity). CTR-CVT is a general term referring to all cardiovascular system damage related to cancer treatment. According to the 2022 ESC Cardio-Oncology Guidelines and domestic expert consensus, it covers 10 major categories of cardiovascular complications: cancer treatment-related cardiac dysfunction (CTRCD), coronary artery disease (CAD), valvular heart disease, arrhythmia, hypertension, thrombosis and thromboembolic diseases, peripheral artery disease and stroke, bleeding complications, pulmonary hypertension, and pericardial disease. CTR-CVT emphasizes the overall damage to the cardiovascular system. CTRCD is a component of CTR-CVT, specifically referring to cardiac dysfunction caused by cancer treatment, including cardiac injury, cardiomyopathy, and heart failure. The ESC Cardio-Oncology Guidelines define CTRCD as the adverse effects of anti-tumor drugs on the cardiac structure and function of cancer patients, manifesting as asymptomatic cardiac dysfunction or symptomatic heart failure, clinically divided into symptomatic CTRCD and asymptomatic CTRCD. For diagnosing asymptomatic CTRCD, in addition to relying on the presence of heart failure-related symptoms (such as worsening shortness of breath after exertion, chest tightness at rest, and paroxysmal nocturnal dyspnea), the internationally recognized main diagnostic criteria currently include a ≥10% decrease in left ventricular ejection fraction (LVEF) from baseline, with the LVEF remaining ≥50% after the decrease, and a mild elevation in serum cardiac function markers (such as troponin and BNP). Because asymptomatic CTRCD, symptomatic CTRCD, and CTR-CVT represent different stages and severity of doxorubicin cardiotoxicity, drugs that can treat or prevent CTR-CVT may not be effective in treating a confirmed CTRCD.

[0005] Doxorubicin cardiotoxicity is a broad term encompassing all cardiac damage that ultimately leads to left ventricular systolic dysfunction and heart failure. Its core target cells are cardiomyocytes, and the main mechanisms include oxidative stress and mitochondrial dysfunction, cardiomyocyte apoptosis and necrosis, calcium homeostasis dysregulation, and energy metabolism crisis, leading to cardiomyocyte death and abnormal systolic and diastolic function, ultimately resulting in cardiomyopathy and heart failure. Doxorubicin Induced Endotheliotoxicity is a subtype of doxorubicin cardiotoxicity (He H, Wang L, Qiao Y, Zhou Q, Li H, Chen S, Yin D, Huang Q, He M. Doxorubicin Induces Endotheliotoxicity and MitochondrialDysfunction via ROS / eNOS / NO Pathway. Front Pharmacol. 2020 Jan 10;10:1531.doi: 10.3389 / fphar.2019.01531, Bosman M, Krüger D, Van Assche C, Boen H, Neutel C, Favere K, Franssen C, Martinet W, Roth L, De Meyer GRY, Cillero-Pastor B, Delrue L, Heggermont W, Van Craenenbroeck EM, Guns PJ. Doxorubicin-induced cardiovascular toxicity: a longitudinal Evaluation of functional and molecular markers. Cardiovasc Res. 2023 Nov 25;119(15):2579-2590. doi:10.1093 / cvr / cvad136.), clinically manifested as asymptomatic CTRCD; its core target cells are cardiac vascular endothelial cells, which can lead to insufficient blood and oxygen supply to the myocardium, indirectly causing myocardial ischemia, hypoxia and dysfunction. There are differences between the two. For example, dexrazoxane, as a cardioprotective agent against anthracycline chemotherapy drugs, mainly protects myocardial cells by chelating iron ions and inhibiting oxygen free radicals. There is insufficient evidence of its direct protective effect on the endothelium. Since the target cells and mechanisms of the two are different, the selectivity of the drugs is different. Therefore, drugs that can treat doxorubicin cardiotoxicity may not be used to treat doxorubicin cardiac endothelial toxicity.

[0006] Currently, there is no literature reporting the effect of Di'ao Xinxuekang on doxorubicin-induced cardiac endothelial damage toxicity. Cardiac endothelial toxicity leads to increased vascular permeability, increasing the contact dose between doxorubicin and cardiomyocytes. At the same time, endothelial toxicity further weakens the nutritional and supportive effects of endothelial cells on cardiomyocytes, affecting cardiomyocyte survival and systolic and diastolic functions. In the early stage, it may manifest as asymptomatic cardiac dysfunction. Currently, there is also no literature reporting Di'ao Xinxuekang in the treatment of tumor treatment-related asymptomatic cardiac dysfunction (CTRCD). Summary of the Invention

[0007] This invention provides a new use for Di'ao Xinxuekang.

[0008] This invention provides the use of Di'ao Xinxuekang in the preparation of a medicament for preventing doxorubicin-induced cardiac endothelial injury toxicity.

[0009] The aforementioned drug is a drug for the prevention and / or intervention of asymptomatic cardiac dysfunction related to cancer treatment.

[0010] The drug is prepared into a commonly used pharmaceutical formulation by adding pharmaceutically acceptable excipients to Di'ao Xinxuekang as the active ingredient.

[0011] The preparation in question is an oral preparation.

[0012] Compared with the literature: Chen Hong, et al., Study on the protective effect and mechanism of Di'ao Xinxuekang on myocardial ischemia-reperfusion injury, Chinese Traditional and Herbal Drugs, Vol. 41, No. 12, December 2010, the literature reported that Di'ao Xinxuekang can improve vascular endothelial function. The literature disclosed an H2O2 injury model in which HUVECs originated from the fetal umbilical vein. Their phenotype, secretion of vasoactive substances, and injury response pathways are fundamentally different from those of adult cardiac vascular endothelial cells, such as coronary artery endothelial cells and cardiac microvascular endothelial cells. In addition, hydrogen peroxide only simulates simple oxidative stress injury, while doxorubicin-induced cardiac vascular endothelial injury involves multiple mechanisms such as DNA damage, non-oxidative apoptosis, and vasodilatory dysfunction. The types of injury are completely different between the two.

[0013] The beneficial effects of this invention are:

[0014] This invention is an unexpected discovery made by the inventors during experimental research on the intervention of doxorubicin-induced cardiotoxicity with Di'ao Xinxuekang. Di'ao Xinxuekang is only effective when administered prophylactically, and its therapeutic effect is not significant. This invention utilizes prophylactic administration of Di'ao Xinxuekang to protect against early cardiac endothelial damage induced by doxorubicin, thereby rescuing or preventing subsequent irreversible cardiotoxicity. In particular, it opens up new applications and methods of administration for the clinical use of Di'ao Xinxuekang by using it to prevent and / or intervene in asymptomatic cardiac dysfunction related to cancer treatment. Attached Figure Description

[0015] Figure 1 Effects of DXXK on cardiac ultrastructure (A), representative CD31 images (B), and percentage of positive area (C) in mice after a single high-dose DOX chemotherapy (Note: Compared with the Control group, *, p<0.05; **, p<0.01. Compared with the DOX group, # p<0.05; ## (p<0.01);

[0016] Figure 2 Effects of DXXK on cardiac ultrastructure (A), representative CD31 images (B), percentage of positive area (C), echocardiography at W4 and W11, and serum marker NT-proBNP results (D, E) in mice subjected to multiple low-dose DOX chemotherapy (Note: Compared with the Control group, *, p<0.05; **, p<0.01. Compared with the DOX group, #, p<0.05; ##, p<0.01).

[0017] Figure 3 Effects of DXXK on cardiac ultrastructure (A), CD31 representative images and percentage of positive area (B), DHE staining representative images and ROS levels (C) in mice receiving multiple doses of conventional DOX chemotherapy (Note: Compared with the Control group, *, p<0.05; **, p<0.01. Compared with the DOX group, # p<0.05; ## (p<0.01);

[0018] Figure 4 Effects of DXXK on cardiac ultrastructure (A), representative CD31 images (B), representative echocardiographic images (C), percentage of CD31-positive area (D), cardiac function (E, F), and myocardial enzymes (G, H) in mice treated with multiple doses of conventional DOX chemotherapy (Note: Compared with the Control group, *, p<0.05; **, p<0.01. Compared with the DOX group, # p<0.05; # (p<0.01). Detailed Implementation

[0019] Experimental Example 1: Di'ao Xinxuekang can improve doxorubicin-induced cardiac endothelial injury.

[0020] To evaluate the protective effect of DXXK (Di'ao Xinxuekang) against DOX-induced cardiac endothelial injury, we established three DOX-induced cardiac endothelial toxicity models to simulate clinical single high-dose DOX chemotherapy, multiple low-dose DOX chemotherapy sessions, and multiple conventional-dose DOX chemotherapy sessions inducing cardiac endothelial injury in mice. Furthermore, to more comprehensively clarify the progression characteristics of the models and the effects of DXXK, this experiment extended the follow-up observation period of the model mice after chemotherapy. The model mice experienced cardiac endothelial injury and asymptomatic cardiac dysfunction, as well as eventual symptomatic cardiac dysfunction. This experiment evaluates the effect of DXXK on DOX-induced early endothelial toxicity and asymptomatic cardiac dysfunction.

[0021] like Figure 1 As shown, in a mouse model of doxorubicin endothelial toxicity induced by a single intraperitoneal injection of DOX 10 mg / kg, on day 3 (D3) after modeling, the pores of endothelial cell junctions enlarged, tight junction proteins became blurred and decreased in number, cell membranes shrank and bulged into the lumen, accompanied by local tissue edema. Similar damage was still observed on days 5 and 20. Compared with the model group, the DXXK group had a more intact endothelial junction structure, a greater number of tight junctions, and well-preserved cell membrane morphology. Figure 1 A). Immunohistochemistry ( Figure 1 B and C show that CD31 expression was significantly decreased at D3, D5, and D20 in the model group (p<0.01), while CD31 expression was significantly increased at the corresponding time points in the DXXK group (p<0.01), suggesting that DXXK can significantly improve endothelial toxicity induced by a single high-dose doxorubicin chemotherapy.

[0022] like Figure 2 As shown, in cumulative intraperitoneal injections of DOX 9 mg / kg (3 mg / kg / week, for 3 consecutive weeks, simulating a clinical low-dose doxorubicin chemotherapy regimen), one week after completing the DOX chemotherapy course (W4), the tight junction proteins in the endothelial cells of the model group animals were blurred and reduced in number, the cell membranes were wrinkled and papillary or triangular protrusions appeared, and the cell nuclei were irregular in shape. Figure 2 A), the expression of the cardiac endothelial marker CD31 was significantly decreased (p<0.01, Figure 2 (B, C), and at the same time, the serum NT-proBNP level in the model animals was significantly increased (p<0.05, Figure 2 D), but ultrasound results in small animals showed no significant changes in left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (p>0.05). Figure 2 By W11, the model mice showed a significant decrease in LVEF and LVFS (p<0.05). Figure 2E) suggests that in the early stage after chemotherapy (W1), the model mice developed clinically asymptomatic cardiac dysfunction. With the extension of follow-up time (W11), the model animals further developed symptomatic cardiac dysfunction. After DXXK intervention, in W4, the cardiac endothelial tight junctions were more abundant, the membrane structure was intact, the protrusions were reduced, and CD31 expression and serum NT-proBNP levels were reversed (p<0.01 or p<0.05). In W11, it could also reverse the decrease in LVEF in the model animals. It suggests that DXXK can significantly protect against the endothelial toxicity of low-dose doxorubicin chemotherapy-induced cardiac dysfunction in mice, improve the early damage of asymptomatic cardiac dysfunction, and avoid the subsequent late-onset cardiac dysfunction.

[0023] like Figure 3 As shown, in the cumulative intraperitoneal injection of DOX 18 mg / kg (3 mg / kg / week, for 6 consecutive weeks, simulating the clinical standard dose of doxorubicin chemotherapy), when the cumulative DOX dose reached 6 mg / kg, the endothelial junction pores enlarged, the tight junction proteins became blurred, the number of papillary or triangular protrusions increased, and edema and exudation appeared in the surrounding tissues. The DXXK group had a more intact endothelial structure, clear tight junctions, and fewer protrusions. Figure 3 A), the expression of the cardiac endothelial marker CD31 was significantly decreased (p<0.05, Figure 3 B), cardiac ROS levels were significantly elevated (p<0.01, Figure 3 (C) With the accumulation of DOX dose, the above-mentioned damage was further aggravated; the above changes were reversed after concurrent DXXK intervention (p<0.01). In summary, the results of the three models consistently indicate that DXXK can effectively protect against DOX-induced cardiac endothelial toxicity.

[0024] Trial Example 2: Prophylactic administration of Di'ao Xinxuekang was effective, while therapeutic administration was ineffective.

[0025] This study compares asymptomatic and symptomatic cardiac dysfunction related to cancer treatment with that treated with Di'ao Xinxuekang.

[0026] Male C57BL / 6 mice were acclimatized for one week and then randomly assigned by body weight to the control group (Control), model group (DOX), DXXK pre-administered group (pDXXK), and DXXK treatment group (tDXXK).

[0027] Except for the blank control group, mice in all other groups were intraperitoneally injected with 3 mg / kg DOX solution on the day of grouping, once a week for 6 weeks. Mice in the blank control group were intraperitoneally injected with the corresponding volume of physiological saline during the same period, with an administration volume of 0.2 ml / 10g.

[0028] The pDXXK and tDXXK groups were administered 320 mg / kg of DXXK solution by gavage daily, starting one week before DOX injection and again seven weeks after injection, until week 12. Symptomatic cardiac dysfunction was induced by week 7 after injection. The control and model groups were administered 320 mg / kg of normal saline by gavage daily, with a gavage volume of 0.1 mL / kg.

[0029] The results showed that when DOX accumulated to 6 mg / kg, mice exhibited elevated serum myocardial enzymes, cardiac vascular endothelial damage, and decreased expression of the endothelial marker CD31 (p<0.01). At a cumulative DOX level of 12 mg / kg, some cardiomyocytes showed mitochondrial swelling and deformation, and the myocardial fiber structure became blurred. At 18 mg / kg, cardiomyocytes showed widespread mitochondrial edema, and the myocardial fiber structure dissolved and disappeared. Figure 4 A, B, and D). Simultaneously, the model mice showed significant abnormalities in various cardiac function indicators (p<0.01). Figure 4 C, E, F). After DOX accumulation reached 18 mg / kg, modeling was stopped and mice were followed up for 6 weeks. Serum myocardial enzymes gradually recovered (p>0.05). Figure 4 Although the damage to the cardiac ultrastructure continued to worsen (G, H), cardiac function remained significantly lower than that of the control group (p<0.01). Figure 4 A), suggesting the irreversibility of chronic myocardial injury. Prophylactic administration of DXXK (320 mg / kg) significantly counteracted changes in the ultrastructure of the cardiac vascular endothelium and decreased CD31 expression in the model animals (p<0.05). Figure 4 A, B, and D), reduce myocardial enzyme levels (p<0.05 or p<0.01, Figure 4 G, H), and reversed late-stage myocardial damage and cardiac dysfunction (p<0.01, Figure 4 A, C, E, F); while therapeutic administration of DXXK had no significant effect, and even with extended treatment for 6 weeks, cardiac function did not show significant improvement (p>0.05). Figure 4 E, F). After 6 weeks of observation following the withdrawal of DOX, the serum myocardial enzymes and body weight of the mice gradually recovered, but the cardiac dysfunction could not be reversed, suggesting that the symptomatic cardiac dysfunction induced by doxorubicin is irreversible. Compared with the reported literature "Deng Liangyan, et al. Study on the protective effect of Di'ao Xinxuekang on doxorubicin-induced cardiotoxicity in zebrafish, Chinese Traditional and Herbal Medicine, 2025;16(2)", this invention observed that after mice developed symptomatic cardiac dysfunction, therapeutic DXXK administration intervention was performed, and it was found that the symptomatic cardiac dysfunction could not be reversed; at the same time, the experiment found that prophylactic administration of DXXK could improve the early cardiac endothelial damage induced by doxorubicin in mice and avoid the subsequent irreversible cardiotoxicity in the model animals, suggesting the application value of prophylactic administration of DXXK for early endothelial toxicity and asymptomatic cardiac dysfunction caused by doxorubicin.

Claims

1. The use of Di'aoxinxuekang in the preparation of a drug for preventing the cardiotoxicity of adriamycin, characterized in that, The doxorubicin cardiotoxicity is tumor treatment related asymptomatic cardiac dysfunction.

2. Use according to claim 1, characterized in that: The medicine is prepared by taking diaoxinexuekang as active ingredient and adding pharmaceutically acceptable adjuvant to prepare the commonly used preparation in pharmacy.

3. Use according to claim 2, characterized in that: The preparation is oral preparation.

Images