Application of VO-OHpic in treatment of myocardial infarction

Abstract

The invention discloses an application of a VO-OHpic trihydrate in preparation of a medicine for preventing and / or treating myocardial infarction. The research finds that the VO-OHpic trihydrate can promote angiogenesis of a myocardial infarction region of a mouse, so that the cardiac fibrosis degree is reduced, and the cardiac function after myocardial infarction is further improved. The research result shows that the VO-OHpic trihydrate can be used for preparing the medicine for preventing and / or treating myocardial infarction, especially the medicine for promoting angiogenesis after myocardial infarction.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to the application of VO-OHpic in the treatment of myocardial infarction, particularly the application of VO-OHpic trihydrate in the preparation of drugs for the prevention and / or treatment of myocardial infarction, such as the preparation of drugs that promote cardiac angiogenesis. Background Technology

[0002] Myocardial infarction leads to cardiac fibrosis and the death and loss of a large number of cardiomyocytes and vascular endothelial cells, resulting in impaired cardiac contractility, left ventricular dilation, scar formation, and thinning of the ventricular wall, ultimately leading to heart failure (Martin, TP et al., RUNX1: an emerging therapeutic target for cardiovascular disease. Cardiovasc Res 116, 1410-1423, doi:10.1093 / cvr / cvaa034(2020)). Meanwhile, due to the significant loss of blood vessels in the infarcted area, there is a lack of blood perfusion, which prevents the effective delivery of blood and oxygen to the damaged cardiac area, ultimately leading to further aggravation of the disease (Huang, M. et al., Endothelial plasticity drives aberrant vascularization and impedes cardiac repair after myocardial infarction. Nat Cardiovasc Res 1,372-388, doi:10.1038 / s44161-022-00047-3(2022)). However, the angiogenesis capacity after myocardial infarction is limited, and the infarcted area cannot generate enough new blood vessels to effectively deliver blood and oxygen to the damaged cardiac area (Kocijan, T. et al., Genetic lineage tracing reveals poor angiogenic potential of cardiac endothelial cells. Cardiovasc Res 117,256-270, doi:10.1093 / cvr / cvaa012(2021)). Currently, there is still a lack of effective treatments for promoting angiogenesis after myocardial infarction to deliver blood and oxygen to the infarcted area and prevent further deterioration of myocardial infarction.

[0003] VO-OHpic trihydrate (CAS No.: 476310-60-8, referred to as VO-OHpic in this article) is a potent and specific PTEN (phosphatase and tensin homolog) inhibitor. Currently, there are no reports on the effects of VO-OHpic on angiogenesis in the infarcted area of ​​myocardial infarction and its further impact on cardiac function. Summary of the Invention

[0004] The technical problem this invention aims to solve is to overcome the deficiency in the prior art of lacking compounds that can effectively promote angiogenesis in the infarct area to prevent and / or treat myocardial infarction, and to provide an application of VO-OHpic in the treatment of myocardial infarction. This invention demonstrates through experiments that VO-OHpic trihydrate can effectively promote angiogenesis after myocardial infarction, thereby reducing cardiac fibrosis and further improving cardiac function after myocardial infarction.

[0005] The present invention solves the above-mentioned technical problems through the following technical solutions:

[0006] A first aspect of the present invention provides the use of a compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a composition comprising the thereof in the preparation of a medicament for the prevention and / or treatment of myocardial infarction.

[0007] The compound is composed of A complex formed with ligand L, wherein ligand L is Ring A is a 5-6 membered heteroaromatic ring with one, two, or three heteroatoms selected from N, O, and S. In the ligand L, -COOH and -OH are located on two adjacent carbon atoms, respectively.

[0008] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier and / or excipients.

[0009] In some embodiments, in the ligand L, the heteroatom of the ring A is N, and the number of heteroatoms is one or two in a six-membered heteroaromatic ring, such as a pyridine ring.

[0010] In some implementations, the ligand L is

[0011] In some embodiments, the compound is composed of A complex consisting of two ligands L.

[0012] In some specific embodiments, the compound is

[0013] In some embodiments, the solvate is a hydrate, such as a trihydrate.

[0014] In some embodiments, the solvate is VO-OHpic trihydrate.

[0015] In some embodiments, the VO-OHpic trihydrate is the VO-OHpic trihydrate shown in CAS No.:476310-60-8.

[0016] In some embodiments, the prevention and / or treatment of myocardial infarction is to promote angiogenesis in the infarcted area by promoting the proliferation of cardiac wall cells and vascular endothelial cells in the infarcted area, thereby preventing and / or treating myocardial infarction.

[0017] In some implementations, the myocardial infarction occurs in mammals.

[0018] In some implementations, the mammal is preferably a human.

[0019] A second aspect of the invention provides the use of a compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a composition comprising the thereof in the preparation of a medicament for promoting angiogenesis.

[0020] The compound, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, or composition comprising the compound as described in the first aspect.

[0021] In some implementations, the promotion of angiogenesis can improve cardiac contractile function and / or reduce the area of ​​cardiac fibrosis and / or increase vascular density in the infarct area.

[0022] In some implementations, the angiogenesis refers to angiogenesis following myocardial infarction.

[0023] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier and / or excipients.

[0024] A third aspect of the invention provides the use of a compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a composition comprising the thereof in the preparation of a medicament for the prevention and / or treatment of heart failure and / or myocardial ischemia.

[0025] The compound, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, or composition comprising the compound as described in the first aspect.

[0026] In some implementations, the heart failure and / or myocardial ischemia are caused by the death and loss of a large number of cardiomyocytes, vascular endothelial cells, and blood vessels.

[0027] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier and / or excipients.

[0028] In some implementations, the dosage form of the drug includes any pharmaceutically acceptable dosage form, e.g. Such as injections, tablets, powders, granules, capsules, oral liquids, ointments, and creams.

[0029] In some implementations, the drug is an injectable form.

[0030] The positive and progressive effects of this invention are as follows:

[0031] This invention demonstrates through experiments that VO-OHpic can effectively promote angiogenesis after myocardial infarction, thereby reducing cardiac fibrosis and further improving cardiac function after myocardial infarction. This invention provides a novel method for treating myocardial infarction and promoting angiogenesis, broadening the application field of VO-OHpic. Therefore, VO-OHpic can be used to prepare drugs for the prevention and / or treatment of myocardial infarction, providing a new drug and treatment approach for the prevention and / or treatment of myocardial infarction. Attached Figure Description

[0032] Figure 1 The effect of VO-OHpic treatment on the cell density of cardiac parietal cells and vascular endothelial cells. Following myocardial infarction, mouse hearts treated with or without VO-OHpic were co-stained with GFP and CD31 antibodies. Scale bar: 100 μm. The relative densities of GFP+ parietal cells and CD31+ endothelial cells in the heart were also statistically analyzed. n = 3 in each group. *** indicates p < 0.001.

[0033] Figure 2 This figure illustrates the effect of VO-OHpic treatment on the cell proliferation of cardiac parietal cells and vascular endothelial cells. Mouse hearts with myocardial infarction treated or untreated with VO-OHpic were co-stained with pHH3 and GFP antibody, or co-stained with pHH3 and CD31 antibody. Arrows indicate proliferating cells. Scale bar is 100 μm. The figure shows the proportion of proliferating GFP+ parietal cells and CD31+ endothelial cells in the infarcted area of ​​mouse hearts with myocardial infarction treated or untreated with VO-OHpic. n = 3 in each group. ** indicates p < 0.01; *** indicates p < 0.001.

[0034] Figure 3 The effect of VO-OHpic treatment on cardiac function is shown in the figure. Echocardiography was used to evaluate cardiac function in mice treated with or without VO-OHpic after myocardial infarction. NS (non-significant), not significant. *p<0.05.

[0035] Figure 4 This figure illustrates the effect of VO-OHpic treatment on cardiac fibrosis. Sirius red staining shows the area of ​​cardiac fibrosis in mouse hearts treated with or without VO-OHpic after myocardial infarction. Scale bar: 2 mm. The figure shows the proportion of cardiac fibrosis area in mouse hearts treated with or without VO-OHpic after myocardial infarction. n = 3 in each group. * indicates p < 0.05. Detailed Implementation

[0036] The embodiments of the present invention will now be described in detail with reference to examples. It should be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention.

[0037] The terms used in this invention are explained as follows:

[0038] The term "solvate" refers to a substance formed by the combination of a compound and a solvent (including but not limited to water, methanol, ethanol, etc.). Solvates are classified into stoichiometric solvates and non-stoichiometric solvates.

[0039] The term "solvate of a pharmaceutically acceptable salt" refers to a substance formed by the combination of a compound with a pharmaceutically acceptable acid or base and a solvent. The amount of solvent can be stoichiometric or non-stoichiometric.

[0040] The term "heteroaromatic ring" refers to a cyclic aromatic ring having a specified number of ring atoms (e.g., 5-6), a specified number of heteroatoms (e.g., 1, 2 or 3), and a specified type of heteroatoms (1, 2 or 3 of N, O and S).

[0041] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0042] The experimental reagents and materials used in the specific embodiments of this invention are as follows:

[0043] Eight to ten-week-old female C57BL / 6 mice and NG2-CreER and Ai47 (NG2, neural / glial antigen 2) genotype mice were used. NG2-CreER mice were purchased from GemPharmatech, and Ai47 mice were obtained from the laboratory of Qiu Zilong at the Songjiang Research Institute of Shanghai Jiao Tong University School of Medicine. (The Ai47 mice were constructed in Qiu Zilong's laboratory; the specific construction method can be found in the literature: Li et al., Generation of a whole-brain atlas for the cholinergic system and mesoscopic projectome analysis of basal forebrain cholinergic neurons. Proceedings of the National Academy of Sciences of Sciences,2018,115(2):415-420), The two mice were crossed to obtain the double-positive NG2. -CreER; Ai47 mice were used for the experiment.

[0044] VO-OHpic (CAS: 476310-60-8, Selleck, catalog number: S8174). Primary antibodies: anti-CD31 antibody (BDBiosciences, catalog number: 553370), anti-pHH3 antibody (Millipore, catalog number: 06-570), and anti-GFP antibody (Abcam, catalog number: ab6662); Secondary antibodies: Donkey anti-goat488 (Invitrogen, catalog number: A-11055), HRP-donkey-anti-rat IgG (Jackson ImmunoResearch, catalog number: 712-035-153), and Donkey anti-rabbit 555 (Invitrogen, catalog number: A-31572).

[0045] Preparation of VO-OHpic solution: Dissolve VO-OHpic powder in DMSO to prepare a stock solution with a final concentration of 50 μg / μL, and then dilute it with physiological saline to prepare a working solution with a concentration of 1 μg / μL.

[0046] Example 1

[0047] (1) Each NG2-CreER;Ai47 mouse was induced to label parietal cells in the heart with 6 mg of Tamoxifen, and myocardial infarction surgery was performed 7 days later. The specific steps of the mouse myocardial infarction surgery are as follows:

[0048] First, the mice were anesthetized with isoflurane in an anesthesia machine. After complete anesthesia, an endotracheal tube was inserted, and the mice were ventilated at a rate of approximately 100 breaths per minute with the assistance of a ventilator to maintain anesthesia at 1-2% isoflurane concentration. Next, the thorax was opened in the intercostal space between the 3rd and 4th ribs on the left side of the chest to expose the mouse's heart. The left anterior descending coronary artery was permanently ligated with 8-0 suture. The pleural cavity and skin were then sutured together with 5-0 suture, and the wound was disinfected with povidone-iodine. Finally, the mice were placed on a 37°C heating plate to keep them warm until they recovered.

[0049] One day before myocardial infarction surgery, each mouse was injected intraperitoneally with either VO-OHpic (at a dose of 10 μg of VO-OHpic per gram of body weight, treatment group) or 200 μL of physiological saline (control group).

[0050] (2) After myocardial infarction surgery, mice were injected intraperitoneally daily with VO-OHpic (at a dose of 10 μg of VO-OHpic per gram of body weight, treatment group) or 200 μL of physiological saline (control group) until 7 days after myocardial infarction surgery.

[0051] (3) Seven days after myocardial infarction surgery, mouse hearts were collected, fixed with 4% paraformaldehyde, dehydrated with 30% sucrose solution, embedded in OCT and then frozen sectioned.

[0052] (4) Soak the slides with tissue sections in PBS for 5 min to remove OCT, then incubate with 3% H2O2 for 15 min each time, repeating 3 times. After that, block with PBST containing 5% normal donkey serum at room temperature for 30 min.

[0053] (5) The primary antibodies were diluted with PBST containing 2.5% normal donkey serum at the corresponding ratios (dilution ratio for anti-GFP antibody: 1:200, for anti-CD31 antibody: 1:500, for anti-pHH3 antibody: 1:1000); the diluted primary antibodies were added to a glass slide with tissue sections attached, and then incubated for 8-12 hours. Then the secondary antibodies were diluted with PBST containing 2.5% normal donkey serum at the corresponding ratios (dilution ratio for Donkey anti-goat 488: 1:1000, for HRP-donkey-anti-rat IgG: 1:50, and for Donkey anti-rabbit 555: 1:1000); the diluted secondary antibodies were added to a glass slide with tissue sections attached, and then incubated at room temperature in a humidified chamber for 60 minutes.

[0054] (6) Prepare TSA colorimetric diluent (30% H2O2:0.1M Borate = 1:10000). Dilute the TSA colorimetric solution with the TSA colorimetric diluent at a volume ratio of 1:1000 and develop the color at room temperature in the dark for 5-10 minutes.

[0055] (7) DAPI (5 mg / mL) was diluted with PBST at a ratio of 1:1000, added to a glass slide, and incubated at room temperature in a humidified chamber for 10 min.

[0056] (8) Add an appropriate amount of fluorescent mounting medium to the slide and seal it. Finally, take a picture using a Zeiss LSM800 laser confocal microscope.

[0057] Example results are as follows Figure 1 and Figure 2 As shown, after myocardial infarction, compared with the control group, treatment with VO-OHpic effectively promoted the proliferation of cardiac parietal cells and vascular endothelial cells, and increased the cell density of parietal cells and vascular endothelial cells, thereby improving the density of coronary vessels.

[0058] Example 2

[0059] (1) One day before myocardial infarction surgery, C57BL / 6 mice were injected intraperitoneally with VO-OHpic (at a dose of 10 μg of VO-OHpic per gram of body weight, treatment group) or 200 μL of physiological saline (control group).

[0060] (2) After myocardial infarction surgery, mice were injected intraperitoneally daily with VO-OHpic (at a dose per gram of body weight). 10μg VO- OHpic dosage) or 200 μL of normal saline, until 2 days after myocardial infarction surgery. 8 days.

[0061] (3) Twenty-eight days after myocardial infarction surgery, echocardiograms were acquired using an ultrasound imaging system (Vevo 2100 Ultrasound, FujiFilm Visual Sonics) to analyze cardiac function in mice. Hearts were then collected for Sirius red staining.

[0062] Example results are as follows Figure 3 As shown, Figure 3 The data presented demonstrated the effects of VO-OHpic treatment on cardiac function. Echocardiography evaluated cardiac function in mice treated with or without VO-OHpic after myocardial infarction (post-MI). NS (non-significant) indicates no statistical difference. * indicates p < 0.05. The results showed that, compared with the control group, VO-OHpic treatment significantly improved cardiac function after myocardial infarction, specifically in terms of ejection fraction and fractional shortening.

[0063] Example 3

[0064] (1) Twenty-eight days after myocardial infarction surgery, mouse hearts were collected, fixed with 4% paraformaldehyde, dehydrated with 30% sucrose solution, embedded with OCT, and then frozen sectioned.

[0065] (2) Soak the slides with the tissue attached in PBS to remove OCT, fix them with 4% paraformaldehyde solution at room temperature for 15 min, and then soak them in PBS for 15 min.

[0066] (3) Following the Sirius red staining procedure, the tissue sections were sequentially passed through Bouin's solution, 0.1% Fast green solution, 1% acetic acid solution, and 0.1% Sirius red solution, and then the slides were immersed in tap water for cleaning.

[0067] (4) Next, the tissue sections were dehydrated by passing them through 95% ethanol, anhydrous ethanol, and xylene in sequence; finally, they were mounted with neutral resin and photographed using an Olympus stereomicroscope (Olympus MVX10).

[0068] Example results are as follows Figure 4 As shown, after myocardial infarction, treatment with VO-OHpic effectively reduced the degree of cardiac fibrosis compared with the control group.

[0069] The above examples demonstrate that treatment with VO-OHpic after myocardial infarction can effectively promote cardiac angiogenesis, reduce the degree of cardiac fibrosis, and further improve cardiac function after myocardial infarction.

Claims

1. The use of a compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a composition comprising the thereof in the preparation of a medicament for the prevention and / or treatment of myocardial infarction; The compound is composed of A complex formed with ligand L, wherein ligand L is Ring A is a 5-6 membered heteroaromatic ring with one, two, or three heteroatoms selected from N, O, and S. In the ligand L, -COOH and -OH are located on two adjacent carbon atoms, respectively.

2. The application as described in claim 1, characterized in that, The composition further comprises a pharmaceutically acceptable carrier and / or excipient; and / or, in the ligand L, the heteroatom of ring A is N, and the number of heteroatoms is one or two, forming a six-membered heteroaromatic ring, such as a pyridine ring; Preferably, the ligand L is 3. The application as described in claim 1 or 2, characterized in that, The compound is composed of A complex consisting of two ligands L; Preferably, the compound is 4. The application as described in any one of claims 1-3, characterized in that, The solvate is a hydrate, such as a trihydrate; Preferably, the solvate is VO-OHpic trihydrate; the VO-OHpic trihydrate is, for example, CAS No.:476310-60-8.

5. The application as described in any one of claims 1-4, characterized in that, The prevention and / or treatment of myocardial infarction is achieved by promoting the proliferation of cardiac wall cells and vascular endothelial cells in the infarct area, thereby promoting angiogenesis in the infarct area, in order to prevent and / or treat myocardial infarction. Preferably, the myocardial infarction occurs in mammals; the mammal is preferably a human.

6. The use of a compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a composition comprising the thereof in the preparation of a medicament for promoting angiogenesis; The compound, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, or a composition comprising the compound as described in any one of claims 1-5.

7. The application according to claim 6, characterized in that, The angiogenesis mentioned refers to angiogenesis following myocardial infarction; And / or, the composition may further comprise a pharmaceutically acceptable carrier and / or excipients.

8. The use of a compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a composition comprising the thereof in the preparation of a medicament for the prevention and / or treatment of heart failure and / or myocardial ischemia; The compound, its pharmaceutically acceptable salt, its solvate, its pharmaceutically acceptable salt solvate, or a composition comprising the compound as described in any one of claims 1-5.

9. The application according to claim 8, characterized in that, The heart failure and / or myocardial ischemia are caused by the death and loss of a large number of cardiomyocytes, vascular endothelial cells and blood vessels; And / or, the composition may further comprise a pharmaceutically acceptable carrier and / or excipients.

10. The application according to any one of claims 1-9, characterized in that, The dosage form of the drug includes any pharmaceutically acceptable dosage form, such as injections, tablets, powders, granules, capsules, oral liquids, ointments, and creams; preferably, the drug is an injection.

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