Application of SGLT2 inhibitor in prevention and treatment of thoracic aortic aneurysm / dissection

Abstract

The invention relates to application of an SGLT2 inhibitor in prevention and treatment of thoracic aortic aneurysm / dissection. Preferably, the SLGT2 inhibitor is empagliflozin, and the SLGT2 inhibitor is SLGT2. The medicine is an oral administration type medicine.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to an application of an SGLT2 inhibitor in preventing and treating thoracic aortic aneurysm / dissection. Background Art

[0002] Aortic aneurysm and aortic dissection are common critical cardiovascular diseases. With the aging of the population and the continuous improvement of imaging diagnostic technology, the incidence rate and the number of patients detected have been on the rise year after year, seriously threatening human life and health. Although thoracic aortic aneurysm / dissection (TAAD) and abdominal aortic aneurysm (AAA) are both diseases characterized by localized dilatation of the aorta, there are significant differences between the two. In addition to the differences in anatomical locations, TAAD and AAA also have their own characteristics in terms of pathology and hemodynamics, and their risk factors and pathogenesis are also different. The causes of TAAD include genetic diseases, congenital diseases, multifactorial degenerative diseases, inflammatory diseases, and infectious diseases. The genetic correlation of TAAD was significantly higher than AAA , about 20% of patients with thoracic aortic aneurysm / dissection have a family history. Atherosclerosis is considered the most common cause of abdominal aortic aneurysm, but the incidence of aortic calcification or atherosclerosis is low in aortic root aneurysms and ascending thoracic aortic aneurysms. In addition, among the more mature animal models of the disease, the classic mouse model of AAA requires the use of apolipoprotein E (ApoE) or low-density lipoprotein (LDL) knockout mice in combination with angiotensin II. These two types of mice are also commonly used mice to construct atherosclerosis models. Angiotensin II has the effects of increasing blood pressure and inducing inflammation. The use of elastase perfusion or calcium chloride application to the abdominal aorta below the renal artery is also a method of establishing abdominal aortic aneurysm. The construction of the TAAD mouse model is significantly different from that of AAA; currently, one of the commonly used models is a gene mutant mouse, such as the Marfan syndrome mouse with a mutation in the Fbn1 gene or the Leoys-Dietz syndrome mouse with a mutation in TGFβ or its receptor, and the other is to give 3-week-old mice the lysyl oxidase inhibitor BAPN for 3-4 weeks. The aortic aneurysm or dissection formed mainly occurs in the ascending aorta, aortic arch and descending aorta, and rarely occurs in the abdominal aorta.

[0003] Among the treatment options for thoracic aortic aneurysm / dissection, surgery and endovascular repair are the main treatment methods, but they require a high level of surgeons and medical institutions. Current drug treatments are mainly for pain relief, lowering blood pressure and heart rate, and reducing the chance of aortic rupture. For aneurysms that have not yet reached surgical indications or non-complex Stanford B clips There is a lack of relatively specific drugs for prevention of leukemia and risk groups. .

[0004] Sodium-glucose cotransporter 2 (SGLT-2) inhibitors are a new type of hypoglycemic drug. SGLT-2 is mainly distributed in the S1 segment of the proximal tubules of the kidneys, and 90% of the glucose reabsorption in the glomerular filtrate is completed by SGLT-2. Inhibition of SGLT-2 can lower blood sugar by reducing glucose reabsorption. In addition, SGLT-2 inhibitors can also inhibit the reabsorption of sodium ions by the proximal tubules, reduce blood volume, thereby reducing cardiac preload and lowering blood pressure. Currently, the therapeutic and / or preventive effects of SGLT-2 in cardiovascular diseases such as atherosclerosis, acute myocardial infarction, type 1 / type 2 diabetes, acute and chronic heart failure have been reported, but there is still no research to prove it in thoracic aortic aneurysm / dissection.

[0005] Based on this, the present invention is proposed. Summary of the invention

[0006] The present invention firstly relates to the use of a SLGT2 inhibitor in preparing a medicine for preventing and / or treating a thoracic aortic aneurysm / dissection.

[0007] Preferably, the SLGT2 inhibitor is empagliflozin;

[0008] The medicine is an oral administration type medicine.

[0009] The beneficial effects of the present invention are:

[0010] The inventors verified for the first time the therapeutic and preventive effects of SLGT2 inhibitors (empagliflozin) in thoracic aortic aneurysm / dissection (TAAD) using animal models and found that oral administration of SGLT2 inhibitors has a good preventive and / or therapeutic effect on thoracic aortic aneurysm / dissection. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 , the effect of preventive administration of SGLT-2 inhibitor (empagliflozin, EMPA) on the survival rate of the mouse thoracic aortic aneurysm / dissection model.

[0012] Figure 2 , the effect of preventive administration of SGLT-2 inhibitor (empagliflozin, EMPA) on the formation and rupture rate of thoracic aortic aneurysm / dissection model in mice. One part of the animals in the drug-treated group did not develop TAAD.

[0013] Figure 3 , The gross image of the aorta shows the preventive effect of SGLT-2 inhibitor (empagliflozin, EMPA) on thoracic aortic aneurysm / dissection in mice: the dissection formation and rupture rates of animals in the preventive drug administration group were reduced.

[0014] Figure 4, aortic EVG staining showed the preventive effect of SGLT-2 inhibitor (empagliflozin, EMPA) on thoracic aortic aneurysm / dissection in mice: the elastic fiber disorder and breakage of animals in the drug-treated group were reduced.

[0015] Figure 5 , and the diameter of the ascending aorta of mice was measured by ultrasound after being fed with BAPN for one week.

[0016] Figure 6 , the inhibitory effect of SGLT-2 inhibitor (empagliflozin, EMPA) on the dilation of the ascending aorta in mice.

[0017] Figure 7 , the effect of SGLT-2 inhibitor (empagliflozin, EMPA) treatment on the rupture rate of thoracic aortic aneurysm / dissection in mice.

[0018] Figure 8 , Aorta gross diagram showing the therapeutic effect of SGLT-2 inhibitor (empagliflozin, EMPA) on thoracic aortic aneurysm / dissection in mice: The occurrence and rupture of dissection were reduced in EMPA-treated mice.

[0019] Fig. 9 , aortic EVG staining showed the therapeutic effect of SGLT-2 inhibitor (empagliflozin, EMPA) on thoracic aortic aneurysm / dissection in mice: the elastic fiber disorder and breakage were reduced in the EMPA therapeutic administration group. DETAILED DESCRIPTION

[0020] Example 1. Preventive effect of SGLT-2 inhibitors on aortic dissection / aortic aneurysm

[0021] The SGLT-2 inhibitor (Empagliflozin) was purchased from Boehringer, specifically named Empagliflozin Tablets, and was mixed into ordinary mouse diet (mouse diet was prepared by Beijing Huafukang Biotechnology Co., Ltd.) at a dose of 300 mg / kg.

[0022] Establishment of TAAD mouse model: 3-week-old male C57BL / 6J mice were taken, 3-aminopropionitrile (BAPN, Sigma-Aldrich, catalog number: A3134) was dissolved in drinking water at a dose of 3 g / L, and the mice were allowed to drink freely for 3 weeks.

[0023] Mouse grouping: TAAD-induced mice (31 mice) were randomly divided into two groups.

[0024] One group was a simple BAPN model (15 mice): the mice were given drinking water containing BAPN and fed with ordinary mouse chow without empagliflozin;

[0025] The other group was the SGLT-2 inhibitor prevention group (16 mice): from the first day of giving drinking water containing BAPN, they were fed with rat diet containing empagliflozin for 3 weeks.

[0026] Observation indicators include:

[0027] 1. Natural death of mice during modeling: record the time of death and cause of death (whether death was caused by dissection / aneurysm rupture);

[0028] 2. At the end of the experiment after 3 weeks of feeding, the surviving mice were killed by an overdose of sodium pentobarbital anesthesia (200 mg / kg body weight), and the blood vessels of all mice were collected for gross photography, and the incidence of thoracic aortic aneurysm / dissection was calculated; the aortic sections were stained for elastic fibers to observe the rupture of elastic plates and the formation of dissections.

[0029] Tissue paraffin section preparation steps:

[0030] (1) At the end of the experiment, mice were anesthetized with pentobarbital (200 mg / kg) and perfused with normal saline to remove residual blood in the heart and aorta.

[0031] (2) The mouse aorta was isolated and cut under a stereomicroscope, and a macroscopic picture was taken, and then the aorta was fixed in 4% paraformaldehyde solution.

[0032] (3) After 24 hours, the ascending aorta was removed and dehydrated, transparentized and paraffin-embedded.

[0033] (4) Embed the tissue and slice it.

[0034] (5) Histopathological examination:

[0035] I. Observe the elastic plate through elastic fiber staining (EVG):

[0036] (1) Paraffin sections are routinely dewaxed with xylene, and then dewaxed to water using different concentrations of ethanol: xylene I (20 min) → xylene II (20 min) → xylene III (20 min) → 100% ethanol (5 min) → 95% ethanol (5 min) → 80% ethanol (5 min) → tap water to wash away the alcohol;

[0037] (2) Prepare elastic fiber staining solution, mix alcohol hematoxylin: ferric chloride: iodine solution in a ratio of 5:2:2, place the slide in the elastic fiber staining solution and leave for 30 minutes;

[0038] (3) Rinse the slide with tap water until there is no floating color;

[0039] (4) Repeated differentiation with differentiation solution, then rinse with tap water and observe the degree of differentiation under a microscope. The elastic fibers appear purple-black and the background appears grayish white and almost colorless.

[0040] (5) Prepare VanGieson dye solution, saturated picric acid and acid fuchsin in a ratio of 9:1, dye for 1-3 minutes, quickly wash with water, remove water, and place in anhydrous ethanol;

[0041] (6) Dehydration with anhydrous alcohol for 5 min;

[0042] (7) Xylene transparent treatment for 5 min and sealing;

[0043] Observed under a Nikon ECLIPSE 90i microscope, elastic fibers appeared black, collagen fibers appeared red, and the background appeared yellow.

[0044] The results show that

[0045] (1) The number of deaths in the EMPA (empagliflozin)-fed group was significantly lower than that in the TAAD model control group during the observation period, and the survival curves showed statistical differences ( Figure 1 ), the rate of interlayer formation and rupture is reduced ( Figure 2 , Figure 3 );

[0046] (2) EVG staining of the ascending aorta showed that the elastic fibers in the TAAD control group were disordered and broken, with dissection and intramural thrombosis. The elastic fibers in the EMPA group were less disordered and broken ( Figure 4 ). The results showed that the SGLT2 inhibitor empagliflozin can inhibit the formation and rupture of thoracic aortic aneurysm / dissection.

[0047] Example 2: Therapeutic effect of SGLT-2 inhibitors on aortic dissection / aortic aneurysm

[0048] 1. Mouse model testing

[0049] The mice were given BAPN and water according to the method described in Example 1. C57BL / 6J mice (16 mice) were randomly divided into two groups.

[0050] One group was the blank control group (6 mice): given drinking water without BAPN;

[0051] One group was the BAPN drinking water group (10 rats): the rats were given BAPN drinking water, and the ascending aorta diameter was measured by aortic ultrasound after 1 week.

[0052] The results showed that after 1 week of BAPN administration, the diameter of the ascending aorta of mice increased compared with the control group, and the difference was statistically significant ( Figure 5 ), indicating that aortic dilatation had occurred in BAPN mice after drinking water for 1 week, which could be used as the starting point for subsequent treatment.

[0053] 2. The therapeutic effect of SGLT-2 inhibitor (empagliflozin) on aortic dissection / aortic aneurysm

[0054] The TAAD mouse model was established using the aforementioned BAPN water feeding method, and mouse diet containing empagliflozin was prepared at the same dose as in Example 1.

[0055] The TAAD-induced mice (36 mice) were randomly divided into two groups.

[0056] One group was the simple BAPN model group (21 mice): BAPN was given drinking water and fed with ordinary mouse chow without empagliflozin;

[0057] The other group was the SGLT-2 inhibitor treatment group (15 mice): starting from one week after being given drinking water containing BAPN, they were fed with rat diet containing empagliflozin.

[0058] Observation indicators:

[0059] 1. At the end of the three-week BAPN experiment, aortic ultrasound was performed to measure the ascending aorta diameter.

[0060] 2. At the end of the experiment, the surviving mice were killed by anesthesia with an overdose of sodium pentobarbital (200 mg / kg body weight), and the blood vessels of all mice were collected for gross photography; the aorta sections were stained for elastic fibers to observe the rupture and dissection of the elastic plates.

[0061] The results show that

[0062] (1) The occurrence and rupture of TAAD model mice in the EMPA treatment group were reduced compared with the TAAD model control group ( Figure 7 , Figure 8 );

[0063] (2) EVG staining of the ascending aorta showed that the elastic fibers of the mice in the TAAD model control group were disordered and broken, with dissection and intramural thrombosis. The elastic fiber disorder and breakage were reduced in the EMPA treatment group ( Fig. 9 ).

[0064] It shows that administering SGLT2 inhibitors (empagliflozin) after thoracic aortic dilatation can inhibit the progression of thoracic aortic aneurysm / dissection, reduce rupture, and stabilize the aortic structure.

Claims

1. Use of SLGT2 inhibitors in the preparation of drugs for preventing and / or treating thoracic aortic aneurysm / dissection.

2. The use according to claim 1, characterized in that: The SLGT2 inhibitor is empagliflozin.

3. The use according to claim 1 or 2, characterized in that: The medicine is an oral administration type medicine.

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