Use of sirt6 activators in the preparation of medicaments for the treatment and prevention of vascular aging-related diseases

Abstract

The application provides application of a SIRT6 activator as an active ingredient in preparation of a medicament for treating and preventing a blood vessel aging related disease. Application of the SIRT6 activator in a blood vessel smooth muscle cell can significantly inhibit aging of the blood vessel smooth muscle cell and increase of an aging related inflammatory factor. In a mouse thoracic aortic aneurysm model constructed by a CaCl2 incubation method, application of the SIRT6 activator can obviously inhibit an aneurysm phenotype.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to the treatment of thoracic aortic aneurysm. BACKGROUND

[0002] Thoracic aortic aneurysm (TAA) is a tumor-like lesion mainly involving the thoracic aortic segment. There are mainly three diseases affecting the normal physiological function of the aorta: atherosclerosis, aneurysm and arterial dissection, the latter two of which are the main culprits causing acute aortic syndrome. Thoracic aortic aneurysm is often called a silent killer because it often has no symptoms, and once the aneurysm ruptures, the mortality rate is very high. At least 22% of people with acute aortic syndrome (including aneurysm or dissection) have died before going to the hospital for treatment, and there is no good treatment measure except surgery after admission. Even these data do not fully reflect the incidence and mortality of thoracic aortic aneurysm, because the onset of thoracic aortic aneurysm is very insidious, and the cause of sudden death is unknown. Therefore, it is particularly important to find possible prevention / treatment measures for thoracic aortic aneurysm.

[0003] Previous studies have shown that familial TAA is mainly caused by genetic factors such as specific gene mutations, such as Marfan syndrome caused by mutation of fibrillin 1 gene, and Loeys-Dietz syndrome caused by mutation of TGFbR1 / TGFbR2 gene. This part of the familial thoracic aortic aneurysm accounts for about 21% of the total thoracic aortic aneurysm. For sporadic thoracic aortic aneurysm, the age of onset of this part of patients is late, and the cause and pathogenesis are not clear. The age of onset of familial thoracic aortic aneurysm is about 56.8 years old, and the average age of patients with sporadic thoracic aortic aneurysm is about 64.3 years old, suggesting that old age may be a risk factor for the onset of sporadic thoracic aortic aneurysm. With the increase of age, the rupture rate of thoracic aortic aneurysm also increases significantly. Studies have shown that for every 10-year increase in age, the rupture rate of thoracic aortic aneurysm increases by 2.6 times. And this risk factor that increases with age is basically unrelated to gender. It is suggested that old age is one of the independent risk factors for the rupture of thoracic aortic aneurysm.

[0004] Cellular senescence is a state of permanent cell cycle arrest that is induced by various stresses, including oncogenic stress. Cellular senescence is a tumor suppressor mechanism that is also involved in other non-neoplastic pathologies. Cells isolated from human atherosclerotic plaques enter senescence more rapidly than normal cells in vitro, and senescent endothelial and smooth muscle cells are present in plaques at the tissue level, as determined by staining for senescence-associated β-galactosidase (SA-βGal). These cells are mainly located in the intima and are associated with the continuous division of cells in pathological conditions. In vitro experiments have shown that senescent vascular smooth muscle cells have a reduced response to NO and to stimulation of β-adrenergic receptors, thus contributing to the reduced vasodilatory activity in elderly individuals. Senescent smooth muscle cells have increased elastase production, in contrast to fibroblasts, and are more prone to apoptosis. Senescent smooth muscle cells can induce chronic inflammation of the vessel wall by secreting inflammation-related factors, thus contributing to the development of atherosclerosis. Recombinant lamin A can cause persistent DNA damage signaling, leading to increased senescence and inflammation secretion in smooth muscle cells. Senescent smooth muscle cells can be involved in the development of age-related vascular diseases. Our published research has shown that vascular smooth muscle cell senescence plays an important role in the development of thoracic aortic aneurysms.

[0005] SIRT6 is a member of the NAD+-dependent class III histone deacetylase Sirtuins (SIRT1-7) family. SIRT6 is closely related to aging and longevity. In 2006, Mostoslavsky et al. first reported that SIRT6 knockout mice exhibited obvious signs of premature aging, such as gray hair, scoliosis, decreased bone density, hypoglycemia, and cachexia. In 2012, it was found that SIRT6 can activate the IGF1 signaling pathway, and SIRT6 transgenic mice have a significantly longer lifespan than wild-type mice.

[0006] The role of SIRT6 in blood vessels has also gradually attracted attention in recent years. A study in 2016 showed that SIRT6 can inhibit the occurrence of mouse atherosclerosis by protecting the function of endothelial cells. And large-scale population sequencing of Sirt6 gene shows that genetic mutations of SIRT6 (rs107251, rs352493, rs3760908) are related to the severity of coronary artery disease. Our research group published a research in July 2023 that proved that SIRT6 knockout can cause vascular smooth muscle cell aging in mice, and aggravate the phenotype of thoracic aortic aneurysm. However, the article only used a mouse thoracic aortic aneurysm model induced by angiotensin AngII to prove that SIRT6 knockout can aggravate thoracic aortic aneurysm, but thoracic aortic aneurysm includes multiple types, and it is unknown whether SIRT6 has an effect on other types of thoracic aortic aneurysm. Moreover, the article does not solve the problem of the application of SIRT6. Can SIRT6 activators inhibit the occurrence of thoracic aortic aneurysm? Can SIRT6 activators be used to treat thoracic aortic aneurysm in other mouse models? These questions are currently unknown.

[0007] In summary, can SIRT6 activators be used as a potential drug for treating thoracic aortic aneurysm? SUMMARY

[0008] To solve the above problems, the present application provides the use of SIRT6 activators as active ingredients in the preparation of a drug for treating and preventing blood vessel aging-related diseases.

[0009] Among them, the blood vessel aging-related disease is thoracic aortic aneurysm, abdominal aortic aneurysm, other aneurysms, and other aging-related vascular diseases.

[0010] Preferably, the blood vessel aging-related disease is thoracic aortic aneurysm. More preferably, the blood vessel aging-related disease is thoracic aortic aneurysm with down-regulated SIRT6 expression.

[0011] Among them, the drug can also include pharmaceutically or immunologically acceptable excipients, carriers and / or diluents.

[0012] Among them, SIRT6 activators include but are not limited to chemical synthetic compounds such as MDL800, MDL811, NAD+ precursor related compounds, etc., natural plant extracts such as icariin, quercetin, etc., and marketed clinical drugs such as SGLT2 inhibitors, GLP-1 receptor agonists, etc.

[0013] Among them, the dosage form of the drug can be oral, suppository, injection or coating, etc.

[0014] The application can significantly inhibit the aging of vascular smooth muscle cells and the increase of aging-related inflammatory factors in human vascular smooth muscle cells by applying SIRT6 activators. In the mouse thoracic aortic aneurysm model constructed by CaCl2 incubation method (in which SIRT6 expression is not down-regulated), the application of SIRT6 activators can obviously inhibit the aneurysm phenotype. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 Shown is the SA-b-gal staining in human vascular smooth muscle cells induced to age by angiotensin AngII, in human SMCs with or without SIRT6 activator MDL-800. Blue indicates SA-b-gal positive cells. The quantification of positive cells is shown on the right. Data are expressed as mean ± SEM.

[0016] Figure 2 Shown is the real-time PCR of aging-related markers and inflammatory factors in human vascular smooth muscle cells induced to age by angiotensin AngIII, with or without SIRT6 activator MDL-800. Data are expressed as mean ± SEM.

[0017] Figure 3 Shown is the SA-b-gal staining in human vascular smooth muscle cells induced to age by multiple passages, in human vascular smooth muscle cells with or without SIRT6 activator MDL-800. Blue indicates SA-b-gal positive cells. The quantification of positive cells is shown on the right. Data are expressed as mean ± SEM. P10 indicates passage to the 10th generation.

[0018] Figure 4 Sirt6 expression in CaCl2 mouse thoracic aortic aneurysm model and control.

[0019] Figure 5 Shown is the CaCl2-induced mouse thoracic aortic aneurysm model incubated with SIRT6 activator MDL-800.

[0020] Figure 6 Shown is the CaCl2-induced mouse thoracic aortic aneurysm model incubated with SIRT6 activator NR. DETAILED DESCRIPTION

[0021] The following examples are used to illustrate the present application, but are not intended to limit the scope of the present application.

[0022] This study was approved by the Medical Ethics Committee of Peking Union Medical College Hospital (PUMCH) and complied with the ethical guidelines, and all participants signed the informed consent form.

[0023] The experimental methods not specifically described in the examples are all conventional experimental methods in the art.

[0024] Example 1 SIRT6 activator can inhibit vascular smooth muscle cell senescence

[0025] In vivo experiments in mice showed that after specific knockout of SIRT6 in smooth muscle, it could significantly cause senescence of vascular medial smooth muscle cells and increase secretion of inflammatory factors of vascular smooth muscle cells. Can SIRT6 activator inhibit smooth muscle senescence to inhibit vascular smooth muscle senescence? We selected SIRT6 specific activator MDL-800 to conduct in vitro experiments.

[0026] We used angiotensin II to induce cell senescence in human primary vascular smooth muscle cells, and at the same time incubated with SIRT6 specific activator MDL-800 (20uM, dissolved in DMSO), MDL-800 is a chemical compound that can bind to the surface allosteric site of SIRT6 to promote SIRT6 deacetylase and demyristoylase activity. SA-βGal staining found that SIRT6 activator can significantly inhibit the senescence of human vascular smooth muscle cells( Figure 1 ). At the same time, fluorescence quantitative PCR showed that SIRT6 activator can significantly inhibit the increase of p53, p21, and the expression of senescence-related inflammatory factors( Figure 2 ).

[0027] In addition to angiotensin II-induced senescence, we also used multiple passages to induce vascular smooth muscle cell senescence. We passaged human vascular smooth muscle cells multiple times to induce senescence (8-9 passages or more), and divided them into two groups during multiple passages, one group was incubated with SIRT6 specific activator MDL-800 (20uM, dissolved in DMSO), and the other group was added with DMSO control. The results showed that SIRT6 specific activator MDL-800 can significantly inhibit multiple passage-induced vascular smooth muscle cell senescence( Figure 3 ).

[0028] These results show that SIRT6 activator can significantly inhibit vascular smooth muscle cell senescence.

[0029] Example 2 SIRT6 activator can inhibit thoracic aortic aneurysm occurrence and disease progression

[0030] We used CaCl2 incubation method to construct thoracic aortic aneurysm model (sponge soaked in 0.5 mol / L CaCl2 solution was applied to the mouse ascending aorta / aortic arch for 15 minutes, and the open chest surgery mouse needed to be assisted with a respirator, and after 4 weeks, thoracic aortic aneurysm formation was observed, and SIRT6 expression was not down-regulated in this model( Figure 4 ). CaCl2 incubation is a direct stimulation of blood vessels, causing calcification of elastic fibers, directly leading to aneurysm, which is a strong and direct stimulation, and SIRT6 cannot react completely in vivo.

[0031] We found that SIRT6 specific activator MDL-800 (50mg / kg, dissolved in DMSO) can significantly inhibit thoracic aortic aneurysm formation in mice (p<0.05) (Fig. 2B). Figure 5

[0032] In addition, we used another SIRT6 activator, Nicotinamide Riboside (NR), in the same experiment. NR is a derivative of vitamin B3, which is the precursor substrate of Nicotinamide Adenine Dinucleotide (NAD+). After intake, it can increase the level of NAD+ to promote the deacetylase activity of SIRT6. We used CaCl2 incubation method to construct thoracic aortic aneurysm model (sponge soaked in 0.5 mol / L CaCl2 solution was applied to the ascending aorta / aortic arch of mice for 15 minutes, and the mice with open chest surgery needed to be assisted with a respirator, and thoracic aortic aneurysm formation was observed after 4 weeks). At the same time, we used subcutaneous slow-release pump method to give SIRT6 activator NR (Sigma Aldrich, 200mg / kg, dissolved in saline) and saline control to mice, and found that SIRT6 activator NR can also significantly inhibit thoracic aortic aneurysm formation in mice (p<0.05) (Fig. 2C). Figure 6

[0033] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the technical principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.​​

Claims

1. The application of SIRT6 activator as an active ingredient in the preparation of drugs for the treatment and prevention of vascular aging-related diseases, wherein the vascular aging-related diseases are thoracic aortic aneurysms with non-SIRT6 downregulated expression, and wherein the SIRT6 activator is MDL-800 or nicotinamide ribose.

2. The application as described in claim 1, characterized in that, The drug also includes pharmaceutically or immunologically acceptable excipients.

3. The application as described in claim 1, characterized in that, The drug also includes pharmaceutically or immunologically acceptable carriers and / or diluents.

4. The application as described in claim 1, characterized in that, The drug dosage forms include oral preparations, suppositories, injections, or topical applications.

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