Application of simetinib in preparation of medicine for promoting myocardial infarction lateral branch artery generation

Abstract

The invention relates to an application of simetinib in preparation of a medicine for promoting myocardial infarction lateral branch artery generation. The invention finds that the application of the simetinib in the early stage of myocardial infarction can improve the cardiac function of a subject after myocardial infarction and promote the formation of the lateral branch artery of the subject after myocardial infarction.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to the use of selumetinib in the preparation of a drug for promoting the formation of collateral arteries after myocardial infarction. Background Art

[0002] At present, the treatment of myocardial infarction is mainly carried out through heart stents and other means, but many elderly or weak people cannot undergo stent surgery. After a myocardial infarction, the coronary arteries of a person's heart can compensate for ischemia by forming collateral arteries, but the ability to form collaterals varies among individuals, and not everyone can form collaterals.

[0003] Currently, there is a lack of drugs that can promote the formation of coronary artery collaterals after myocardial infarction.

[0004] Selumetinib is an anticancer drug, and there has never been any report that selumetinib can promote the formation of collateral arteries. Summary of the invention

[0005] Based on this, one embodiment of the present invention provides the use of selumetinib in the preparation of a drug for promoting the formation of collateral arteries after myocardial infarction.

[0006] The technical solutions include the following:

[0007] In a first aspect, the present application provides the use of selumetinib in the preparation of a drug for promoting the formation of collateral arteries after myocardial infarction.

[0008] In some embodiments, the promoting of collateral artery formation after myocardial infarction includes increasing the density of arteries in the infarct area after myocardial infarction and / or increasing the density of collateral arteries in the infarct junction area after myocardial infarction.

[0009] In a second aspect, the present application provides the use of selumetinib in the preparation of a drug for improving cardiac function after myocardial infarction.

[0010] In a third aspect, the present application provides the use of selumetinib in the preparation of a drug for alleviating myocardial fibrosis after myocardial infarction.

[0011] In a fourth aspect, the present application provides the use of selumetinib in the preparation of a drug for promoting the transformation of the macrophage M1-M2 phenotype.

[0012] In a fifth aspect, the present application provides the use of selumetinib in the preparation of a drug for preventing or treating myocardial infarction.

[0013] In some embodiments, the selumetinib has the efficacy of promoting the increase of the expression of M2 cell markers and inhibiting the decrease of the expression of M1 cell markers.

[0014] In some embodiments, the active ingredient of the drug is selumetinib, and the drug further includes pharmaceutically acceptable excipients.

[0015] In some embodiments, the excipients include, but are not limited to, at least one of a sustained-release agent, a solubilizer, a stabilizer, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, a surfactant, and a lubricant.

[0016] In some embodiments, the excipient includes a vegetable oil, including but not limited to corn oil. The vegetable oil, such as corn oil, can be used as a solubilizer (corn oil, as a hydrophobic solvent, can dissolve those drugs that are insoluble in water, ensuring that the drugs can be evenly distributed and properly dissolved in the injection), a sustained-release agent (due to the high viscosity of corn oil, it can slow down the release rate of the drug in the body, allowing the drug to be gradually absorbed, thereby prolonging the drug's action time. This allows corn oil to act as a sustained-release agent in some cases), an excipient (corn oil is used as a carrier of the drug in intraperitoneal injection to ensure that the drug can be stably present in the body and effectively transported to the site of action), and a stabilizer (corn oil can protect the drug from oxidation or degradation, especially for those drugs that are sensitive to moisture, corn oil can play a certain stabilizing role).

[0017] In some embodiments, the auxiliary material includes DMSO (dimethyl sulfoxide), which can be used as a solubilizer, absorption enhancer, excipient, and stabilizer.

[0018] In some embodiments, the auxiliary material includes PEG300, and PEG300 can be used as a solubilizer, excipient, wetting agent, stabilizer, and absorption enhancer.

[0019] In some embodiments, the auxiliary material includes SBE-β-CD, and SBE-β-CD can be used as a solubilizer, stabilizer, sustained-release agent, and absorption enhancer.

[0020] In some embodiments, the auxiliary material includes Tween-80, which can be used as a surfactant, solubilizer, wetting agent, stabilizer, lubricant, and absorption enhancer.

[0021] In some embodiments, the solvent formula for dissolving selumetinib drug mainly includes the following three:

[0022] (1)10% DMSO+40% PEG300+5% Tween-80+45% Saline;

[0023] (2) 10% DMSO+90% (20% SBE-β-CD in Saline);

[0024] (3) 10% DMSO + 90% Corn Oil.

[0025] In some embodiments, the dosage form of the drug includes injection, tablet, capsule, granule, oral solution, pill or sustained-release agent.

[0026] In some embodiments, the injection is a liquid injection, a powder for injection, or a tablet for injection.

[0027] In some embodiments, the administration route of the injection includes one of intravenous injection, intraperitoneal injection, intramuscular injection or subcutaneous injection.

[0028] In some embodiments, the effective dose of selumetinib treatment is not higher than 5 mg / kg. Optionally, the effective dose of selumetinib treatment is 1 mg / kg to 5 mg / kg, and the optional values ​​include but are not limited to 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, and the range composed of any two values ​​of the above values. Optionally, the effective dose of selumetinib treatment is 2.5 mg / kg to 5 mg / kg. In some other embodiments, the effective dose that can achieve the purpose of treatment by those skilled in the art according to the disclosure of this application and conventional treatment methods is within the scope of protection of this application.

[0029] In some embodiments, the medication method: short-term high-frequency medication can be used in the early stage of myocardial infarction, and low-frequency medication can be used in the late stage of myocardial infarction; in some embodiments, the medication frequency in the early stage of myocardial infarction is once a day, and the continuous medication does not exceed 5 days. Optionally, the medication frequency in the late stage of myocardial infarction is once every 3 to 4 days to reduce the toxic and side effects of selumetinib. In some other embodiments, the medication methods that can achieve the purpose of treatment according to the disclosure of this application and conventional treatment methods by those skilled in the art are all within the scope of protection of this application.

[0030] In some embodiments, the early / pre-myocardial infarction period may refer to the 0th to 3rd day after the occurrence of myocardial infarction, and short-term high frequency refers to once-daily medication for 0-3 days, and the subsequent frequency is reduced to once every 3-4 days (low-frequency medication).

[0031] Optionally, the drug is administered once at a fixed time point, for example, promptly at 9 am every day.

[0032] In some other embodiments, those skilled in the art may adopt other modes of administration. Any modes of administration that can achieve the purpose of treatment by those skilled in the art based on the disclosure of this application and conventional treatment methods are within the protection scope of this application.

[0033] In a sixth aspect, the present application provides a drug for promoting the formation of collateral arteries after myocardial infarction, the active ingredient of which is an effective dose of selumetinib.

[0034] Compared with the traditional technology, this application has the following beneficial effects:

[0035] The present invention finds that the application of selumetinib in the early stage of myocardial infarction can improve the cardiac function of the subject after myocardial infarction and promote the formation of collateral arteries after myocardial infarction. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The results of the ejection fraction (EF) of mice were detected by color Doppler ultrasound of the heart on the 28th day after myocardial infarction after drug administration;

[0037] Figure 2 The results of immunofluorescence staining of frozen sections of mouse hearts on day 28 after myocardial infarction were used to detect a-SMA / CD31 for marking arterial smooth muscle and endothelial cells.

[0038] Figure 3 The results of microCT examination of mouse hearts on the 28th day after myocardial infarction after drug administration;

[0039] Figure 4 The results of Masson staining were obtained from paraffin sections of mouse hearts on the 28th day after myocardial infarction after drug administration;

[0040] Figure 5 Immunofluorescence staining of frozen sections of mouse hearts was performed on the third day after myocardial infarction to detect the results of a-SMA / CD68 / CD206 for marking arterial smooth muscle, macrophages and M2 macrophages respectively;

[0041] Figure 6 Immunofluorescence staining of frozen sections of mouse hearts was performed on day 3 after myocardial infarction to detect the results of a-SMA / CD86 / CD206 for marking arterial smooth muscle, M1 macrophages, and M2 macrophages, respectively;

[0042] Figure 7 The results are shown in Figure 2. The mouse hearts were sampled with liquid nitrogen on the third day after myocardial infarction, and proteins were extracted and then tested by Western Blot. DETAILED DESCRIPTION

[0043] In order to facilitate understanding of the present invention, the present invention will be described more fully below, and the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thorough and comprehensive.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0045] The terms "and / or", "or / and", and "and / or" used in this article have a selection scope that includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, said any and all combinations including any two related listed items, any more related listed items, or a combination of all related listed items.

[0046] Herein, "plurality", "multiple", "multiple times", "plurality", etc., unless otherwise specified, refer to a number greater than or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0047] Herein, “preferred”, “better”, “more preferred” and “suitable” are merely used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute limitations on the scope of protection of the present invention.

[0048] Herein, “further”, “furthermore”, “particularly”, etc. are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present invention.

[0049] In this article, "optionally", "optional", and "optional" mean optional, that is, any one of the two parallel options of "yes" or "no". If there are multiple "options" in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "option" is independent.

[0050] In this article, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0051] Herein, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when a numerical interval only refers to integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all sub-ranges included therein.

[0052] Herein, "effective dose" will vary according to the specific state, disease, disorder or condition being treated and its severity, as well as the age, weight, physical condition and responsiveness of the individual being treated. Therefore, one or more of these parameters can be used to select and adjust the therapeutically effective amount of selumetinib. A person of ordinary skill in the art can routinely adjust and determine the therapeutically effective amount based on their knowledge and the technical solutions disclosed in the present invention.

[0053] As used herein, the term "treatment" may refer to any alleviation, alleviation or elimination of disease symptoms in an individual, alleviation of any direct or indirect pathological consequences of the disease, reduction of the rate of disease progression, alleviation or improvement of the disease state, and alleviation or improvement of prognosis due to the administration of a drug.

[0054] Herein, in the present invention, "subject" refers to a patient taking a drug.

[0055] Herein, "subject" is an animal, preferably a mammal, more preferably a human, and the subject includes but is not limited to a patient with a disease, illness and / or symptom. The subject in the present invention is preferably a mammal. The term "mammal" mainly refers to warm-blooded vertebrate mammals, including but not limited to: such as cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice (such as rats, mice), pigs, cattle, sheep, horses, humans, etc., preferably primates, more preferably humans.

[0056] Selumetinib is mainly used for anti-cancer treatment, and its application dosage is usually 25 mg / ㎡ orally taken twice a day (approximately every 12 hours), and the application cycle is usually one month. Selumetinib has drug toxicity, and long-term use may cause cardiotoxicity. The present invention avoids the toxic effects of high-dose selumetinib by using the drug at a high frequency in the early stage of myocardial infarction (0th, 1st, 2nd, and 3rd days) and at a low frequency in the middle and late stages of myocardial infarction (7th, 10th, and 14th days). The present invention found that short-term high-frequency application of selumetinib in the early stage of myocardial infarction can improve the cardiac function of the subject after myocardial infarction and promote the formation of collateral arteries after myocardial infarction in the subject.

[0057] In addition, selumetinib can inhibit the inflammatory response of the heart in the early stage of myocardial infarction, promote the M1-M2 transformation of macrophages, and thus promote the formation of coronary collaterals.

[0058] The following is a detailed description in conjunction with specific examples. The following examples do not include other components except for inevitable impurities unless otherwise specified. The reagents and instruments used in the examples are conventionally selected in the art unless otherwise specified. The experimental methods for which specific conditions are not specified in the examples are implemented according to conventional conditions, such as the conditions described in the literature, books, or methods recommended by the manufacturer.

[0059] The main experimental reagents involved in the following examples are:

[0060] Selumetinib MCE HY-50706;

[0061] 4% PFA Servicebio G1101-500ML;

[0062] OCT embedding medium Scigen 4586;

[0063] Bovine serum albumin Sigma-Aldrich A4737;

[0064] Anti-Actin, α-smooth muscle-Cy3 TM Antibodies, mouse monoclonal Sigma-Aldrich C6198;

[0065] Human / Mouse / Rat CD31 / PECAM-1Antibody R&D system AF3628;

[0066] CD68Antibody(E-11)santacruz sc-17832;

[0067] CD86(B7-2)Monoclonal Antibody(GL1),eBioscience invitrogen 14-0862-82;

[0068] CD206 / MRC1(E6T5J) Rabbit mAb Cell signaling 24595S;

[0069] Donkey anti-Goat IgG(H+L)Cross-Adsorbed Secondary Antibody,AlexaFluor TM 647Invitrogen TM A-21447;

[0070] Donkey anti-Rabbit IgG(H+L)Highly Cross-Adsorbed Secondary Antibody,Alexa Fluor TM 647Invitrogen TM A-31573;

[0071] Donkey anti-Rat IgG(H+L)Highly Cross-Adsorbed Secondary Antibody,Alexa Fluor TM 488Invitrogen TMA-21208;

[0072] DAPI Fluoromount-G TM Anti-fluorescence quenching mounting medium (containing DAPI, water-soluble) yeasen36308ES11;

[0073] Microfil FlowTech MV-122;

[0074] RIPA lysis buffer Biyuntian P0013B;

[0075] 5x SDS loading buffer Biyuntian P0015;

[0076] Anti-beta Tubulin antibody-Loading Control Abcam ab6046;

[0077] Anti-TNF alpha antibody[EPR19147]Abcam ab183218;

[0078] Anti-IL-10antibody[EPR1114]Abcam ab133575;

[0079] VEGFA Polyclonal antibody proteintech 19003-1-AP;

[0080] Goat Anti-Rabbit IgG H&L(HRP)Abcam ab6721;

[0081] Enhanced ECL Chemiluminescent Substrate Kit yeasen 36222ES60;

[0082] The following embodiments involve the following main methods:

[0083] 1. Ejection fraction detection by cardiac ultrasound

[0084] On the 28th day after myocardial infarction modeling, the mice were subjected to cardiac ultrasound imaging using a high-frequency echocardiography system (VisualSonics Vevo 2100). First, the mice were anesthetized (1.5%-2% isoflurane) and placed on a heating pad to maintain body temperature (37°C). Depilatory cream was used to remove chest hair, and ultrasound gel was applied to the chest to avoid air bubble interference. Left ventricular long-axis section images were obtained using M mode, and the end-diastolic volume (LVEDV) and end-systolic volume (LVESV) were measured. The ejection fraction was calculated according to the formula: EF = [(LVEDV-LVESV) / LVEDV] × 100%. Each mouse was measured at least three times, and the average value was taken for analysis.

[0085] 2. Immunofluorescence staining of cardiac cryosections

[0086] Mouse hearts were collected on the 3rd and 28th days after myocardial infarction modeling. The mice were anesthetized and fixed by cardiac perfusion, and 4% paraformaldehyde (PFA) was perfused through the left ventricle. After the heart tissue was fixed, it was removed and post-fixed in 4% PFA for 12 hours, and then transferred to 30% sucrose solution for dehydration for more than 48 hours. The heart was embedded in OCT (optimal cutting temperature) compound and frozen at -80°C. Heart sections were prepared as continuous sections with a thickness of 10 μm using a freezing microtome. The sections were dried at room temperature for 30 minutes and then washed 3 times with PBS for 5 minutes each time. Blocked with 5% BSA for 1 hour, and then primary antibodies (SMA 1:200, CD31 1:100, CD68 1:100, CD86 1:100, CD206 1:100) were added and incubated overnight at 4°C. The next day, the sections were washed three times with PBS for 5 minutes each time, and then the corresponding fluorescent-labeled secondary antibodies (Donkey antigoat AF647 1:500, Donkey anti rabbit AF647 1:500, Doonkey anti rat AF488 1:500, Doonkey anti mouse AF488 1:500) were added and incubated at room temperature for 1 hour. Finally, the sections were stained with DAPI for nuclei and imaged using a fluorescence microscope (Zeiss LSM 900).

[0087] 3. Mouse Heart MicroCT Detection

[0088] On the 28th day after myocardial infarction modeling, mice were anesthetized (1.5%-2% isoflurane) and perfused with 4% paraformaldehyde (PFA) through the thoracic aorta. After sufficient perfusion, microfil contrast agent was perfused through the thoracic aorta to fully fill the coronary artery with contrast agent. After the contrast agent solidified, the heart was separated and fixed in 4% paraformaldehyde (PFA) for 24 hours. After fixation, the heart was scanned in vitro by microCT (Bruker SkyScan 1276). The sample was placed on the microCT scanning bed, and the scanning parameters were set. The typical settings were voltage 50kV, current 200μA, and scanning resolution 10μm. The 3D image was reconstructed by software (NRecon, CTAn) and the diameter and number of the coronary artery collateral lumen of the heart were measured to evaluate the generation of coronary collateral circulation in the heart.

[0089] 4.Masson staining

[0090] The heart tissue was fixed in 4% paraformaldehyde (PFA), then routinely paraffin-embedded and sliced ​​(4 μm thick). The slices were baked at 58°C for 1 hour to be fixed on glass slides. Using the Masson trichrome staining method, staining pretreatment was first performed, including dewaxing, rehydration and Bougival solution staining of the slices. Collagen fibers were then treated and stained with phosphotungstic acid. Finally, the muscle fibers were stained with acid fuchsin and decolorized with 1% acetic acid. After staining, the slices were dehydrated with ethanol and transparentized with xylene, and then observed and photographed under an optical microscope after sealing.

[0091] 5. Western Blot Detection

[0092] The heart tissue was frozen in liquid nitrogen and ground into powder, and then lysed with RIPA lysis buffer (containing protease and phosphatase inhibitors). After ultrasonic treatment, the samples were centrifuged at 4°C for 15 minutes, and the supernatant was taken to determine the protein concentration. An equal amount of SDS loading buffer was added to each sample, and SDS-PAGE electrophoresis (10% or 12% gel) was performed after boiling for 5 minutes. After electrophoresis, the protein was transferred to a PVDF membrane, and the membrane was blocked in 5% skim milk powder for 1 hour. Primary antibodies (β-Tublin 1:2000, TNF-α1:1000, IL-10 1:1000, VEGF 1:1000) were added and incubated at 4°C overnight. The next day, HRP-labeled secondary antibodies (goat anti rabbit HRP 1:5000) were used for incubation at room temperature for 1 hour, and the expression of the target protein was detected by ECL chemiluminescence. Images were collected by an imaging system (Bio-Rad), and grayscale analysis was performed using ImageJ software.

[0093] The chemical structure of Selumetinib (AZD6244) involved in the following examples is shown below:

[0094]

[0095] Treatment plan: Giving 2.5 mg / kg of selumetinib once at fixed time points (such as 9 am every day) on days 0, 1, 2, 3, 7, 10, and 14 after myocardial infarction can promote the formation of collateral circulation after myocardial infarction.

[0096] Formula: Selumetinib is dissolved in 10% DMSO + 90% Corn Oil to a concentration of 1.25 mg / ml. Among them, each component of the solvent only plays at least one role among a sustained-release agent, a solubilizer, a stabilizer, an excipient, a filler, an adhesive, a wetting agent, a disintegrant, an absorption promoter, a surfactant and a lubricant, and it is selumetinib that exerts the therapeutic effect. The following examples use 10% DMSO + 90% Corn Oil (abbreviated as OC) as a solvent to dissolve selumetinib.

[0097] Example 1 Intraperitoneal injection of selumetinib can promote the formation of collateral circulation after myocardial infarction in mice

[0098] 1. Intraperitoneal injection of selumetinib can improve the cardiac function of mice 28 days after myocardial infarction

[0099] Myocardial infarction modeling was performed in 10-week-old male C57BL / 6 mice. The brief method was as follows: After the mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital, endotracheal intubation was performed and the ventilator was connected. The mice were opened in the chest, the pericardium was separated, the heart was exposed, and the anterior descending coronary artery was ligated with a 10-0 needle suture. After ligation, the local myocardium turned from red to white. The chest cavity of the mice was sutured with 6-0 sutures, and the endotracheal intubation was removed. The EF value of the modeled mice was measured by cardiac color Doppler ultrasound the next day. The range of 20%-40% was considered a successful modeling, and subsequent experiments could be performed.

[0100] On days 0, 1, 2, 3, 7, 10, and 14 after myocardial infarction, mice with myocardial infarction were intraperitoneally injected with 0 (OC), 2.5 mg / kg, and 5 mg / kg of selumetinib. On day 28 after myocardial infarction, the ejection fraction of the mice was detected by color Doppler ultrasound of the heart. The test results are as follows: Figure 1 As shown, it was found that the ejection fraction of mice injected with selumetinib was significantly higher than that of mice injected with OC, among which the ejection fraction of mice injected with 2.5 mg / kg was more significantly increased.

[0101] 2. Intraperitoneal injection of selumetinib can increase the arterial density in the infarct area of ​​mice 28 days after myocardial infarction

[0102] Myocardial infarction mice were intraperitoneally injected with 0 (OC), 2.5 mg / kg, and 5 mg / kg of selumetinib on days 0, 1, 2, 3, 7, 10, and 14 after myocardial infarction. Immunofluorescence staining of frozen sections of mice hearts was performed on day 28 after myocardial infarction to detect a-SMA / CD31 for marking arterial smooth muscle and endothelial cells. The test results are shown in Figure 2 As shown, it was found that the density of a-SMA-positive arterioles per unit field of view in mice injected with selumetinib was significantly higher than that in mice injected with OC, among which the increase in mice injected with 2.5 mg / kg dose of selumetinib was more obvious.

[0103] 3. Intraperitoneal injection of selumetinib can increase the density of collateral arteries in the infarct junction area of ​​mice 28 days after infarction

[0104] The mice with myocardial infarction were intraperitoneally injected with 0 (OC) and 2.5 mg / kg of selumetinib on days 0, 1, 2, 3, 7, 10, and 14 after myocardial infarction, and microCT of the mouse heart was performed on day 28 after myocardial infarction. Figure 3 As shown, the density of arteries in the myocardial infarction junction zone (watershed zone) of mice injected with selumetinib was significantly higher than that of mice injected with OC.

[0105] 4. Intraperitoneal injection of selumetinib can reduce the degree of fibrosis in the infarct area of ​​mice 28 days after myocardial infarction

[0106] Myocardial infarction mice were intraperitoneally injected with 0 (OC) and 2.5 mg / kg of selumetinib on days 0, 1, 2, 3, 7, 10, and 14 after myocardial infarction. Paraffin sections of the mice hearts were collected on day 28 after myocardial infarction and tested with Masson staining. Figure 4 As shown, the extent of myocardial fibrosis was found to be significantly lower in mice injected with selumetinib than in mice injected with OC.

[0107] Example 2 Intraperitoneal injection of selumetinib promotes collateral formation after myocardial infarction by promoting M1-M2 transformation. Current studies have shown that M2 macrophages can promote the formation of collateral arteries after myocardial infarction. Selumetinib injection can promote the formation of collateral arteries after myocardial infarction by promoting the transformation of macrophage M1-M2 phenotypes.

[0108] 1. Intraperitoneal injection of selumetinib can increase the density of M2 macrophages around the arteries in the infarct area of ​​mice 3 days after myocardial infarction

[0109] Myocardial infarction mice were intraperitoneally injected with 0 (OC) and 2.5 mg / kg of selumetinib on days 0, 1, and 2 after myocardial infarction, respectively. Immunofluorescence staining of frozen heart sections was performed on day 3 after myocardial infarction to detect a-SMA / CD68 / CD206, which were used to mark arterial smooth muscle, macrophages, and M2 macrophages, respectively.

[0110] Test results such as Figure 5 As shown, the number of a-SMA-positive periarterial CD68+ / CD206+ macrophages per unit field of view in mice injected with selumetinib was significantly greater than that in mice injected with OC.

[0111] 2. Intraperitoneal injection of selumetinib can reduce the density of M1 macrophages around the arteries in the infarct area of ​​mice 3 days after myocardial infarction

[0112] Myocardial infarction mice were intraperitoneally injected with 0 (OC) and 2.5 mg / kg of selumetinib on days 0, 1, and 2 after myocardial infarction, respectively. Immunofluorescence staining of frozen heart sections was performed on day 3 after myocardial infarction to detect a-SMA / CD86 / CD206, which were used to mark arterial smooth muscle, M1 macrophages, and M2 macrophages, respectively.

[0113] Test results such as Figure 6 As shown, the number of a-SMA-positive periarterial CD86+ / CD206- macrophages per unit field of view in mice injected with selumetinib was significantly lower than that in mice injected with OC.

[0114] 3. Intraperitoneal injection of selumetinib can respectively reduce and increase the expression levels of M1 and M2 markers in mouse heart tissue 3 days after myocardial infarction

[0115] On days 0, 1, and 2 after myocardial infarction, mice with myocardial infarction were intraperitoneally injected with 0 (OC) and 2.5 mg / kg of selumetinib. On day 3 after myocardial infarction, the mouse hearts were sampled with liquid nitrogen, and proteins were extracted and then tested by Western Blot. Figure 7 As shown, the M2 cell markers (IL-10, VEGF) of mice injected with selumetinib were significantly more than those of mice injected with OC, and the M1 cell markers (TNF-a) were significantly less than those of mice injected with OC.

[0116] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0117] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Use of selumetinib in the preparation of a drug, wherein the drug is used for at least one of the following (a) to (d): (a) Promote the formation of collateral arteries after myocardial infarction; (b) Improve cardiac function after myocardial infarction; (c) reduce myocardial fibrosis after myocardial infarction; (d) Promote the transformation of macrophage M1-M2 phenotype.

2. The use according to claim 1, characterized in that: The promoting of collateral artery formation after myocardial infarction includes increasing the density of arteries in the infarcted area after myocardial infarction and / or increasing the density of collateral arteries in the infarcted junction area after myocardial infarction.

3. The use according to claim 1, characterized in that: The selumetinib promotes the transformation of macrophage M1-M2 phenotype, thereby promoting the formation of collateral arteries after myocardial infarction.

4. The use according to any one of claims 1 to 3, characterized in that The active ingredient of the drug is selumetinib, and the drug also includes pharmaceutically acceptable excipients.

5. The use according to claim 4, characterized in that: The auxiliary materials include one or more of sustained-release agents, excipients, solubilizers, stabilizers, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants and lubricants.

6. The use according to any one of claims 1 to 3, characterized in that The dosage form of the drug includes injection, tablet, capsule, granule, oral solution, pill or sustained-release preparation.

7. The use according to claim 6, characterized in that: The administration route of the injection includes one of intravenous injection, intraperitoneal injection, intramuscular injection or subcutaneous injection.

8. The use according to any one of claims 1 to 3, characterized in that The effective dose of selumetinib for treatment is no higher than 5 mg / kg.

9. A drug for promoting the formation of collateral arteries after myocardial infarction, characterized in that: The active ingredient is an effective dose of selumetinib.

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