Use of tamoxifen in preparation of drug for treating heart failure

Abstract

A use of tamoxifen or a pharmaceutically acceptable salt thereof in preparation of a drug for treating heart failure. Pharmacodynamic studies have shown that tamoxifen can significantly reduce the mRNA levels of myocardial hypertrophy indicators and inflammatory factors, improve cardiac functions of mice with heart failure induced by left anterior descending artery ligation, enhance EF and FS values in the mice with heart failure, and can thus be used for treating heart failure, providing a new option for developing a novel anti-heart failure drug.

Description

Application of tamoxifen in preparing medicine for treating heart failure Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to an application of tamoxifen in the preparation of a medicine for treating heart failure. Background Art

[0002] Heart failure is generally considered a process in which cardiomyocytes undergo progressive necrosis, replaced by scar tissue or diffuse interstitial fibrosis, leading to a continuous decrease in functional cardiomyocytes. Symptoms and signs are caused by impairment of any structure or function responsible for ventricular filling or pumping. Currently, drugs used in the clinical treatment of heart failure can be divided into four major categories: renin-angiotensin-aldosterone system inhibitors, beta-blockers, mineralocorticoid receptor antagonists, and sodium-glucose cotransporter-2 inhibitors. In addition, a variety of other medications, such as digoxin and diuretics, are available for patients with heart failure with varying etiologies and symptoms. Heart failure patients are characterized by complex etiologies and numerous comorbidities. Many patients have limited options for heart failure treatment due to medication effects on blood pressure, potassium levels, renal function, and other physical conditions, or due to concurrent medication use. As a growing global public health problem, heart failure urgently requires the exploration of new targets and new drugs.

[0003] Tamoxifen (ICI 46474), molecular formula C 26 H 29NO is a selective estrogen receptor modulator, mainly used to treat estrogen receptor-positive breast cancer, and is also used to treat non-invasive ductal carcinoma. In addition to estrogen receptors, tamoxifen targets also include protein kinase C, metalloproteinase-1, vascular endothelial growth factor, transforming growth factor-β, IL-6, etc., showing multiple activities such as inhibiting the growth, differentiation and metastasis of cancer cells, antagonizing oxidative stress, and inhibiting fibrosis (Ahmed NS, Samec M, Liskova A, Kubatka P, Saso L. Tamoxifen and oxidative stress: an overlooked connection. Discov Oncol. 2021 May 27; 12(1): 17.) (Grainger DJ, Witchell CM, Metcalfe JC. Tamoxifen elevates transforming growth factor-beta and suppresses diet-induced formation of lipid lesions in mouse aorta. Nat Med. 1995 Oct; 1(10): 1067-73.) ( D, Flegar D, M, P, P, A, N, D, Kelava T. Tamoxifen Ameliorates Cholestatic Liver Fibrosis in Mice: Upregulation of TGFβand IL6 Is a Potential Protective Mechanism. Biomedicines. 2022 May 23; 10(5): 1209.). Tamoxifen can reduce the levels of C-reactive protein and fibrinogen (Weitzman J. Tamoxifen is good for the heart. Trends Mol Med. 2001 Apr; 7(4): 150.); and tamoxifen can also improve lipid oxidative damage by regulating the metabolism of low-density lipoprotein (Williams JK, Wagner JD, Li Z, Golden DL, Adams MR. Tamoxifen inhibits arterial accumulation of LDL degradation products and progression of coronary artery atherosclerosis in monkeys. Arterioscler Thromb Vasc Biol. 1997 Feb; 17(2): 403-8.).In several clinical statistics of breast cancer treatment, tamoxifen has a lower risk of cardiovascular disease than other breast cancer treatment drugs such as aromatase inhibitors and gonadotropin-releasing hormone agonists (Yu Q, Xu Y, Yu E, Zheng Z. Risk of cardiovascular disease in breast cancer patients receiving aromatase inhibitors vs. tamoxifen: A systematic review and meta-analysis. J Clin Pharm Ther. 2022 May; 47(5): 575-587.)(Reis SE, Costantino JP, Wickerham DL, Tan-Chiu E, Wang J, Kavanah M. Cardiovascular effects of tamoxifen in women with and without heart disease: breast cancer prevention trial. National Surgical Adjuvant Breast and Bowel Project Breast Cancer Prevention Trial Investigators. J Natl Cancer Inst. 2001 Jan 3; 93(1): 16-21.). In addition, tamoxifen also has the effects of preventing arrhythmias after ischemia-reperfusion in rats (Pavón N, Hernández-Esquivel L, Buelna-Chontal M, Chávez E. Antiarrhythmic effect of tamoxifen on the vulnerability induced by hyperthyroidism to heart ischemia / reperfusion damage. J Steroid Biochem Mol Biol. 2014 Sep; 143: 416-23.) and preventing and treating cardiotoxicity caused by the anesthetic bupivacaine (patent application with publication number CN105726526A, title: Use of tamoxifen in the preparation of a drug for preventing and treating cardiotoxic side effects caused by bupivacaine).

[0004] Patent application publication number KR1020220022300A discloses a dilated cardiomyopathy model and its preparation method, relating to an animal model of dilated cardiomyopathy with a cardiac-specific BIS gene deletion and its preparation method. By controlling the timing of dilated cardiomyopathy induction, cardiomyopathy can be induced within 3 minutes after taking tamoxifen for several weeks. Compared with existing methods, this method shortens the time required to induce dilated cardiomyopathy and is economical. Furthermore, the animal model of the present invention is expected to be used for screening and validating dilated cardiomyopathy drugs. Tamoxifen is used to induce knockout of exon 3 of the BIS gene in mice to construct a heart failure model. To date, there have been no reports on the use of tamoxifen in the treatment of heart failure.

[0005] Summary of the Invention

[0006] The purpose of the present invention is to provide an application of tamoxifen in preparing a medicine for treating heart failure.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] The first aspect of the present invention provides a use of tamoxifen or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating heart failure.

[0009] The chemical structure of tamoxifen (National Center for Biotechnology Information. PubChem Compound Summary for CID 2733526, Tamoxifen. https: / / pubchem.ncbi.nlm.nih.gov / compound / Tamoxifen. Accessed Oct. 13, 2023.) is shown below:

[0010] The pharmaceutically acceptable salt is an acid addition salt formed by tamoxifen and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

[0011] The medicine for treating heart failure also includes pharmaceutically acceptable excipients.

[0012] Common clinical manifestations of heart failure include dyspnea, fatigue, fluid retention, pulmonary edema, cardiogenic shock, hypertension, etc. Clinically, patients with past or current symptoms and / or signs of heart failure caused by abnormal cardiac structure and / or function, and who also meet one of the following criteria: elevated natriuretic peptide levels or objective evidence of pulmonary or systemic congestion obtained through imaging or hemodynamic measurements, are defined as patients with heart failure (Bozkurt B, Coats AJS, Tsutsui H, et al. Universal definition and classification of heart failure: a report of the Heart Failure Society of America, Heart Failure Association of the European Society of Cardiology, Japanese Heart Failure Society and Writing Committee of the Universal Definition of Heart Failure: Endorsed by the Canadian Heart Failure Society, Heart Failure Association of India, Cardiac Society of Australia and New Zealand, and Chinese Heart Failure Association. Eur J Heart Fail. 2021; 23(3): 352-380.). Its pathogenesis involves increased cardiac preload and postload, neuroendocrine system abnormalities, energy metabolism remodeling, cardiac structural remodeling, and myocardial electrical remodeling. The "new quadruple" of drugs currently used clinically to treat heart failure includes renin-angiotensin-aldosterone system inhibitors, beta-blockers, mineralocorticoid receptor antagonists, and sodium-glucose cotransporter-2 inhibitors. In addition, other drugs include guanylate cyclase agonists, digoxin, and diuretics.

[0013] The present invention uses a mouse heart failure model induced by ligating the left anterior descending artery for four weeks (Riehle C, Bauersachs J. Small animal models of heart failure. Cardiovasc Res. 2019 Nov 1; 115(13): 1838-1849.) to observe the anti-heart failure effect of tamoxifen administered orally. This animal model is a widely recognized anti-heart failure model. The pathophysiological evolution process of the established heart failure model is similar to that of humans, and the model effect is good (Huang Ming, Xiong Ke, Li Xiao, et al. Research Progress of Animal Models of Heart Failure [J]. Journal of Tianjin University of Traditional Chinese Medicine, 2019, 38(06): 534-540; Wang Jie, Duan Kunkun, Wang Yafeng. Research Status of Methods for Establishing Animal Models of Heart Failure [J]. Journal of Northwest Minzu University (Natural Science Edition), 2022, 43(01): 52-57.).The cell hypertrophy model used a combination of isoproterenol and phenylephrine to stimulate HL-1 cardiomyocyte hypertrophy (Zhao Shuai, Li Qingling, Han Xu, et al. Research progress of isolated cardiomyocyte injury models [J]. Journal of Changchun University of Chinese Medicine, 2015, 31(03): 650-653; Viereck J, Kumarswamy R, Foinquinos A, Xiao K, Avramopoulos P, Kunz M, Dittrich M, Maetzig T, Zimmer K, Remke J, Just A, Fendrich J, ScherfK, Bolesani E, Schambach A, Weidemann F, Zweigerdt R, de Windt LJ, Engelhardt S, Dandekar T, Batkai S, Thum T. Long noncoding RNA Chast promotes cardiac remodeling. Sci Transl Med. 2016 Feb 17;8(326):326ra22.); the cellular inflammation model used lipopolysaccharide to induce HL-1 cardiomyocyte inflammation (Yang Z, Pan X, Wu X, Lin Q, ChenY, Cai S, Zhang Y, Mai Z, Ahmad N, Ma D, Deng L. TREM-1 induces pyroptosis in cardiomyocytes by activating NLRP3 inflammasome through the SMC4 / NEMO pathway. FEBS J. 2023 Mar; 290(6):1549-1562.), both of which are widely recognized in vitro cell models used to observe the effects of tamoxifen on cardiomyocyte hypertrophy and inflammation in vitro.

[0014] Experimental results showed that after continuous oral administration of 3 mg / kg of tamoxifen for 28 days, the effect of improving ejection fraction (EF%) and short-axis shortening rate (FS%) was basically equivalent to that of 2 mg / kg of Enalapril, a commonly used ACEI drug in clinical practice. The heart volume was significantly reduced and the morphology was significantly improved. Echocardiography showed that the ventricular inner diameter was reduced and the ventricular wall pulsation was enhanced. In HL-1 cells in vitro, 3 μM tamoxifen can improve the mRNA level of the hypertrophic factor brain natriuretic peptide BNP after ISO / PE stimulation and the mRNA levels of the inflammatory factors TNF-α and IL-1b after LPS stimulation, and has the effect of inhibiting myocardial cell hypertrophy and inflammation.

[0015] The second aspect of the present invention provides a use of tamoxifen or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating myocardial hypertrophy.

[0016] The third aspect of the present invention provides a use of tamoxifen or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating myocardial cell inflammation.

[0017] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0018] The present invention provides a new medical use for tamoxifen. Pharmacodynamic studies have shown that tamoxifen can significantly reduce myocardial hypertrophy markers and mRNA levels of inflammatory factors. It also improves cardiac morphology and function in mice with left anterior descending artery ligation-induced heart failure, as well as the EF and FS values ​​of these mice. Tamoxifen can be used to treat heart failure. This invention provides a new option for the search for new anti-heart failure drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] FIG1 is an M-mode echocardiogram of mice after administration of different doses of tamoxifen.

[0020] FIG2 is a schematic diagram showing the effects of different doses of tamoxifen on mouse cardiac morphology.

[0021] FIG3 is a schematic diagram showing the effects of different doses of tamoxifen on the ejection fraction (EF%) of mice with heart failure (means±SEM, n=7-10; *P<0.05, **P<0.01, ***P<0.001).

[0022] FIG4 is a schematic diagram showing the effects of different doses of tamoxifen on fractional shortening (FS%) in mice with heart failure (means±SEM, n=7-10; *P<0.05, **P<0.01, ***P<0.001).

[0023] FIG5 is an M-mode echocardiogram of mice in a 3 mg / kg tamoxifen time-effect experiment.

[0024] FIG6 is a schematic diagram showing the effect of 3 mg / kg tamoxifen on mouse heart morphology in a time-effect experiment.

[0025] Figure 7 is a schematic diagram showing the effects of 3 mg / kg tamoxifen on ejection fraction (EF%) in heart failure mice at 3, 7, 14, and 28 days (means±SEM, n=7-10; *P<0.05, **P<0.01, ***P<0.001).

[0026] Figure 8 is a graph showing the effects of 3 mg / kg tamoxifen on fractional shortening (FS%) in heart failure mice at 3, 7, 14, and 28 days (means±SEM, n=7-10; *P<0.05, **P<0.01, ***P<0.001).

[0027] FIG9 is a schematic diagram showing the effect of 3 μM tamoxifen on the level of BNP mRNA, a hypertrophy indicator in HL-1 cardiomyocytes (means±SEM, n=3; *P<0.05, **P<0.01, ***P<0.001).

[0028] Figure 10 is a schematic diagram showing the effects of 3 μM tamoxifen on the mRNA levels of inflammatory factors TNF-α, IL-6 and IL-1b in HL-1 cardiomyocytes (means±SEM, n=3; *P<0.05, **P<0.01, ***P<0.001). DETAILED DESCRIPTION

[0029] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. Those skilled in the art should understand that the following specific description is illustrative rather than restrictive and should not be used to limit the scope of protection of the present invention.

[0030] Example 1

[0031] In vivo pharmacodynamics study of tamoxifen in mice

[0032] 1.1 Tamoxifen in vivo pharmacodynamics in mice Materials: C57 mice, small animal gas anesthesia machine, MyLab high-frequency ultrasound equipment, SL3116 transducer, depilatory cream, isoflurane, sutures, corn oil, and normal saline.

[0033] 1.2 Left Anterior Descending Artery Ligation Heart Failure Model: Hair on the chest and abdomen of mice was removed using depilatory cream. The thoracotomy was performed at the fourth and fifth intercostal spaces on the left thorax, exposing the heart. A vein running toward the apex of the heart was visible between the left atrial appendage and the conus arteriosus. Deep within this vein was the left anterior descending coronary artery, which was ligated with suture approximately 1 cm below the inferior margin of the left atrial appendage. The chest cavity was then closed and sutured, and the mice were maintained as normal after awakening. Sham-operated (Sham) mice were treated identically to the experimental group except for the lack of ligation. Four weeks after the ligation procedure, echocardiography was performed, with mice anesthetized with isoflurane. Hair on the chest and abdomen of the mice was removed using depilatory cream. Echocardiographic data were recorded using a MyLab high-resolution in vivo microimaging system and an SL3116 transducer in two-dimensional long-axis views. Indicators such as left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) were measured.

[0034] 1.3 Tamoxifen dose-effect study

[0035] 1.3.1 Animal grouping and drug administration

[0036] A successful heart failure model was established, with 20% < EF% < 50%. Mice were randomly divided into eight groups, including a sham operation group (Sham), a heart failure model group (HF), an enalapril group (HF+E, 2 mg / kg), and 0.3, 1, 3, 10, and 30 mg / kg tamoxifen groups (HF+TAM). Tamoxifen was prepared in corn oil and administered orally at a dose of 0.1 mL per 10 g of body weight. Both the Sham and HF groups received the same dose of corn oil once daily for 28 days. Echocardiography was then performed to compare changes in cardiac function before and after tamoxifen administration.

[0037] The results are shown in Figures 1 to 4. Figure 1 shows M-mode echocardiograms of mice after administration of different doses of tamoxifen. As can be seen, ventricular internal diameters decreased and ventricular wall pulsations increased after administration. Figure 2 shows the effects of different doses of tamoxifen on mouse cardiac morphology. As can be seen, the heart volume of the model heart failure mice significantly increased, while enalapril and tamoxifen administration reduced heart volume, resulting in intact heart morphology and a smoother surface. The 3 mg / kg tamoxifen group showed a more pronounced effect than the other dose groups.

[0038] Figure 3 shows the effects of different tamoxifen doses on ejection fraction (EF) in mice with heart failure (means ± SEM, n = 7-10; *P < 0.05, **P < 0.01, ***P < 0.001). Ejection fraction increased in all tamoxifen dose groups compared to pre-dose, with the 3 mg / kg and 10 mg / kg groups achieving improvements equivalent to those achieved with the active drug enalapril.

[0039] Figure 4 shows the effects of different tamoxifen doses on fractional shortening (FS%) in mice with heart failure (means ± SEM, n = 7-10; *P < 0.05, **P < 0.01, ***P < 0.001). Tamoxifen doses increased FS% compared to pre-dose, with the 3 mg / kg group achieving an improvement equivalent to that of the active drug enalapril.

[0040] The experimental results showed that after continuous oral administration of different doses of tamoxifen for 28 days, the effect of 3 mg / kg tamoxifen in improving ejection fraction (EF%) and short axis shortening rate (FS%) was basically equivalent to 2 mg / kg of Enalapril, a commonly used ACEI drug in clinical practice. It reversed heart failure in mice with left anterior descending artery ligation and improved the cardiac function and morphology of mice with heart failure.

[0041] 1.4 3mg / kg Tamoxifen Time-Effect Study

[0042] 1.4.1 Animal grouping and drug administration

[0043] The 3mg / kg tamoxifen group showed an effect equivalent to enalapril in improving heart failure in a dose-effect experiment, and a time-effect experiment was conducted on it. The screened mice were randomly divided into 4 groups, including a sham operation group (Sham), a model group (HF), an enalapril group (HF+E, 2mg / kg), and a tamoxifen group (HF+TAM, 3mg / kg). Tamoxifen was prepared using corn oil and gavage was performed according to the standard of 0.1mL per 10g body weight of the mouse. The Sham group and the HF group were given the same dose of corn oil once a day. Cardiac ultrasound examinations were performed on days 3, 7, 14, and 28 of administration to compare changes in cardiac function before and after tamoxifen administration.

[0044] The results are shown in Figures 5 to 8. Figure 5 shows M-mode echocardiograms of mice in a 3 mg / kg tamoxifen time-effect study. Tamoxifen administration reduced ventricular internal diameter and enhanced ventricular wall pulsation. Figure 6 shows the effects of 3 mg / kg tamoxifen on mouse cardiac morphology in a 3 mg / kg tamoxifen time-effect study. After 3 mg / kg tamoxifen administration, mouse heart volume was significantly reduced, and cardiac morphology remained intact. Figure 7 shows the effects of 3 mg / kg tamoxifen on ejection fraction (EF%) in mice with heart failure at 3, 7, 14, and 28 days (means ± SEM, n = 7-10; *P < 0.05, **P < 0.01, ***P < 0.001). Figure 8 shows the effects of 3 mg / kg tamoxifen on fractional shortening (FS%) in mice with heart failure at 3, 7, 14, and 28 days (means ± SEM, n = 7-10; *P < 0.05, **P < 0.01, ***P < 0.001). The experimental results showed that after continuous oral administration of tamoxifen 3 mg / kg for 0, 7, 14 and 28 days, the effect of improving ejection fraction (EF%) and short axis shortening rate (FS%) was basically equivalent to 2 mg / kg of Enalapril, a commonly used ACEI drug in clinical practice. It reversed heart failure in mice with left anterior descending artery ligation and improved the cardiac function and morphology of mice with heart failure.

[0045] Example 2

[0046] Effect of tamoxifen on ISO / PE combined-induced HL-1 cardiomyocyte hypertrophy model

[0047] Mouse HL-1 cardiomyocytes were stimulated with isoproterenol (ISO, 100 μM) and phenylephrine (PE, 100 μM) for 24 hours to induce hypertrophy. Control, ISO / PE, and tamoxifen (3 μM) treatment groups were established. RT-qPCR was used to detect the mRNA expression of ANP and BNP in this primary cardiomyocyte hypertrophy model.

[0048] Figure 9 shows the effect of 3 μM tamoxifen on BNP mRNA levels, a marker of cardiomyocyte hypertrophy, in HL-1 cells (means ± SEM, n = 3; *P < 0.05, **P < 0.01, ***P < 0.001). BNP is a sensitive marker of impaired cardiac function. The results show that 3 μM tamoxifen can ameliorate the elevated BNP mRNA levels following ISO / PE stimulation, demonstrating an inhibitory effect on cardiomyocyte hypertrophy.

[0049] Example 3

[0050] Effects of tamoxifen in LPS-induced HL-1 cardiomyocyte inflammation model

[0051] Mouse HL-1 cardiomyocytes were stimulated with 10 μg / mL LPS for 24 hours to induce inflammation. Control, LPS-stimulated, and tamoxifen (3 μM) treatment groups were established. RT-qPCR was used to detect the mRNA expression of TNF-α, IL-6, and IL-1b in the primary cardiomyocyte inflammation model.

[0052] Figure 10 shows the effect of 3 μM tamoxifen on mRNA levels of the inflammatory factors TNF-α, IL-6, and IL-1b in HL-1 cardiomyocytes (means ± SEM, n = 3; *P < 0.05, **P < 0.01, ***P < 0.001). The experimental results show that tamoxifen significantly ameliorated the LPS-induced increase in mRNA levels of the pro-inflammatory factors TNF-α and IL-1b, inhibiting cardiomyocyte inflammation.

[0053] In view of the above efficacy test results, the tamoxifen of the present invention has significant anti-heart failure activity and can therefore be used to prepare anti-heart failure drugs or foods.

[0054] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as above in terms of a preferred embodiment, it is not intended to limit the present invention. Any technician familiar with this patent can make slight changes or modifications to equivalent embodiments using the above technical content without departing from the scope of the technical solution of the present invention. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the solution of the present invention.

Claims

1. Use of tamoxifen or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating heart failure.

2. Use of tamoxifen or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a medicament for treating heart failure, characterized in that: The chemical structure of tamoxifen is shown below:

3. Use of tamoxifen or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a medicament for treating heart failure, characterized in that: The pharmaceutically acceptable salt is an acid addition salt formed by tamoxifen and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, lactic acid, citric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, tartaric acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

4. Use of tamoxifen or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating myocardial cell hypertrophy.

5. Use of tamoxifen or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating myocardial cell inflammation.

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