Application of Apilimod in preparation of medicine for preventing or treating myocardial infarction

Abstract

The invention provides an application of Apilimod in preparation of a medicine for preventing or treating myocardial infarction. The Apilimod is a compound shown in a formula (I) or a pharmaceutically acceptable form of the compound. Experiments prove that the Apilimod can obviously relieve the cardiac function after myocardial infarction. The invention provides a new treatment approach for myocardial infarction. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to the use of Apilimod in medicine for preventing or treating myocardial infarction. Background Art

[0002] Myocardial infarction (MI) is a cardiovascular disease caused by an imbalance between blood and oxygen supply and demand in the coronary arteries. Its pathological hallmark is myocardial hypoxia and damage due to restricted blood flow to the heart caused by coronary artery disease. Currently, revascularization procedures such as coronary intervention or coronary artery bypass grafting (CABG) are considered the optimal treatment for patients with MI. However, factors such as postoperative restenosis, reperfusion injury, extensive lesions, and low surgical tolerance limit their efficacy in MI. Therefore, exploring drug treatments has potential clinical value. Currently, commonly used Western medications include statins, angiotensin-converting enzyme inhibitors (ACEIs), beta-blockers (BBs), angiotensin II receptor blockers (ARBs), nitrates, and antiplatelet drugs. However, these drugs are often associated with adverse clinical reactions.

[0003] Apilimod inhibits the synthesis of IL-12 and IL-23 in patients with Crohn's disease (CD), rheumatoid arthritis (RA), and psoriasis. In B-cell non-Hodgkin's lymphoma, Apilimod inhibits tumor cell proliferation through autophagy. There are no reports of studies using this compound in the prevention or treatment of myocardial infarction. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the present invention aims to provide a new drug for preventing or treating myocardial infarction. Based on this purpose, the present invention adopts the following technical solutions:

[0005] In one aspect, the present invention provides the use of a compound represented by formula (I) or a pharmaceutically acceptable form thereof in the preparation of a medicament for preventing or treating myocardial infarction:

[0006]

[0007] As an embodiment, the pharmaceutically acceptable form includes salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs.

[0008] As an embodiment, the drug further includes a pharmaceutically acceptable carrier and / or excipient.

[0009] In the present invention, the pharmaceutically acceptable carriers and / or excipients include but are not limited to solvents, diluents, dispersing aids, suspending aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, or lubricants.

[0010] As an embodiment, the dosage form of the drug includes tablets, capsules, granules, pills, drop pills, syrups, powders, granules, suppositories, drops, emulsions, injections, solutions or suspensions.

[0011] In the present invention, the administration of the drug includes but is not limited to intravenous injection, subcutaneous injection, intramuscular injection, transdermal administration, local administration, implantation, or sustained-release administration.

[0012] As a preferred embodiment, the drug is administered by intravenous injection.

[0013] As an embodiment, the dosage of the drug is 2 mg / kg.

[0014] As an embodiment, the drug is administered once a day.

[0015] According to a specific embodiment of the present invention, the prevention or treatment of myocardial infarction includes alleviating the effects of damage on myocardial cells and promoting the recovery of cardiac function.

[0016] According to a specific embodiment of the present invention, the prevention or treatment of myocardial infarction includes improving the contractile function of the left ventricle, or improving the ejection fraction after myocardial infarction.

[0017] According to a specific embodiment of the present invention, the compound as shown in formula (I) or a pharmaceutically acceptable form thereof can enhance the homeostasis of myocardial cells under injury conditions, reduce myocardial necrosis, and enhance the homeostasis of the endocardium / vascular-myocardial tissue extracellular matrix (HSPG).

[0018] On the other hand, the present invention also provides a method for preventing or treating myocardial infarction, which comprises administering a compound represented by formula (I) or a pharmaceutically acceptable form thereof to a subject.

[0019] In the present invention, the subject includes mammals or humans.

[0020] As an embodiment, the compound or a pharmaceutically acceptable form thereof is administered at a dosage of 2 mg / kg.

[0021] As an embodiment, the administration of the compound or its pharmaceutically acceptable form includes intravenous injection, subcutaneous injection, intramuscular injection, transdermal administration, topical administration, implantation, or sustained-release administration.

[0022] In one embodiment, the compound or a pharmaceutically acceptable form thereof is administered in a single dose.

[0023] On the other hand, the present invention also provides a method for protecting endocardial / vascular-cardiomyocytes and extracellular matrix under injury conditions, comprising administering a compound of formula (I) or a pharmaceutically acceptable form thereof to the cells.

[0024] The present invention has the following beneficial effects:

[0025] 1. Experiments at the molecular and cellular levels in vitro have shown that the Apilimod of the present invention can enhance the endocardial / vascular endothelial-myocardial extracellular matrix HSPG and myocardial cell cTnT under injury conditions, and alleviate the reduction in the number of myocardial cells caused by injury (infarct area, such as Figure 1 The red frame portion shows the effect of protecting myocardial cells. In the present invention, the Apilimod is a compound as shown in formula (I) or a pharmaceutically acceptable form thereof.

[0026] 2. In vivo animal experiments have shown that the administration of the Apilimod of the present invention improves the ejection fraction and left ventricular contractile function of the heart after myocardial infarction, thereby promoting the recovery of cardiac function. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This figure shows the results of detecting the effect of the compound Apilimod on the expression levels of HSPG and cardiomyocyte cTnT in the endocardial / vascular endothelial cell-cardiomyocyte co-culture area exposed to injury conditions;

[0028] Figure 2 These are the results of testing the effect of the compound Apilimod on myocardial infarction, where Figure A is a schematic diagram of medication and testing time points, Figure B is a statistical graph of cardiac ultrasound EF values ​​at different time points, and Figure C is an echocardiogram at different time points. DETAILED DESCRIPTION

[0029] Terms used in this invention:

[0030] The term "prevent" in the context of the present invention refers not only to completely preventing a certain effect, but also to substantially preventing, reducing, lowering, diminishing or eliminating any part of the effect before or in the early stages of the disease.

[0031] The term "treat" in the context of the present invention refers to any effect that has a beneficial effect on the progression of a disease, including attenuating, reducing, diminishing or eliminating the development of pathology after disease onset.

[0032] The term "pharmaceutically acceptable carrier" includes any and all solvents, diluents or other liquid vehicles, dispersion or suspension aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc., suitable for preparing the particular dosage form desired. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol and phosphate buffers, and other nontoxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives, and antioxidants may also be present in the composition, according to the judgment of the formulator.

[0033] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is substantially non-toxic to living organisms. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by reacting a compound of the present invention with a pharmaceutically acceptable inorganic / organic acid or inorganic / organic base. Such salts are also referred to as acid addition salts or base addition salts. Common inorganic acids include (but are not limited to) hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.; common organic acids include (but are not limited to) trifluoroacetic acid, citric acid, maleic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, oxalic acid, formic acid, acetic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.; common inorganic bases include (but are not limited to) sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide, etc.; common organic bases include (but are not limited to) diethylamine, triethylamine, ethambutol, etc.

[0034] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has at least one chiral factor (including chiral center, chiral axis, chiral plane, etc.) resulting in a perpendicular asymmetric plane and can rotate plane polarized light.

[0035] The term "tautomer" (or "tautomeric form") refers to structural isomers with different energies that can be interconverted via a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (or prototropic tautomers) include, but are not limited to, interconversions via proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-iminoalcohol isomerization, and the like. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

[0036] The term "solvate" refers to a substance formed by the combination of a compound of the present invention or a pharmaceutically acceptable salt thereof with at least one solvent molecule through non-covalent intermolecular forces. Common solvates include (but are not limited to) hydrates, ethanolates, acetonides, etc.

[0037] The term "chelate" refers to a complex having a cyclic structure, which is obtained by the chelation of two or more ligands with the same metal ion to form a chelate ring.

[0038] The term "non-covalent complex" is formed by the interaction of a compound with another molecule, wherein no covalent bond is formed between the compound and the molecule. For example, complexation can occur through van der Waals interactions, hydrogen bonding, and electrostatic interactions (also known as ionic bonding).

[0039] The term "prodrug" refers to a derivative compound that is capable of providing, directly or indirectly, a compound of the invention upon administration to a patient. Particularly preferred derivative compounds or prodrugs are those that increase the bioavailability of a compound of the invention when administered to a patient (e.g., are more readily absorbed into the bloodstream) or that enhance the delivery of the parent compound to the site of action.

[0040] Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are well known in the art.

[0041] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing specific embodiments rather than for limiting the scope of protection of the present invention.

[0042] When the embodiments provide numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the understanding of the prior art by those skilled in the art and the description of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention may also be used to implement the present invention.

[0043] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in this technical field.

[0044] Example 1 Protective effect of compound Apilimod on cardiomyocytes at the in vitro cellular level

[0045] Cell culture and drug administration

[0046] In this study, hiCMs (human-induced cardiomyocytes, cardiomyocytes derived from human induced pluripotent stem cells) (greater than 30 days) and hiEnLCs (human-induced endocardial-like cells, endocardial-vascular endothelial-like cells derived from human induced pluripotent stem cells) (passage number less than 4) were co-cultured on a Culture-Insert 2Well microslide (ibidi) with a 0.5mm intercellular gap. Each cell type was coated with Matrigel before inoculation. First, hiCMs were plated at 5.0E+04 / cm 2 Three days later, hiEnLCs treated with 200 nM Apilimod for 30 minutes were seeded onto Matrigel. Twenty-four hours after seeding, all cells were cultured in EGM-2 medium containing Matrigel diluted 1:100 for 1-2 days until both cell types reached 100% confluence. They were then exposed to injury-conditioned medium and fixed and immunofluorescence stained three days later.

[0047] Immunofluorescence staining

[0048] Cells were fixed in 4% paraformaldehyde (PFA) for 15 minutes, washed three times in PBS, washed in PBS for 15 minutes, and cleared in 0.3% Tween-20 / PBS or 0.1% Triton X-100 / PBS (PBST) for 15 minutes, followed by blocking in 3% BSA for 30 minutes. Primary antibodies cTnT (Invitrogen MA5MA5-12960, 1:400) and HSPG (USBiological H1890H1890, 1:200) were incubated overnight at 4°C. Following incubation, cells were washed three times with PBS and incubated with secondary antibodies goat anti-mouse IgG2b-AF647 (Invitrogen A21242, 1:1000) and goat anti-rabbit IgG-AF555 (Invitrogen A32732, 1:1000) for 60 minutes at room temperature. Images were acquired using a PE high-content cell imager (PerkinElmer).

[0049] The results are as follows Figure 1 As shown, the results indicate that the compound Apilimod increases the expression of HSPG and cTnT at the in vitro cellular level, alleviating the effects of damage on cardiomyocytes.

[0050] Example 2 Apilimod's promoting effect on the increase of EF value after myocardial infarction in vivo

[0051] Test methods

[0052] mice

[0053] C57BL / 6J mice were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd., Sibeifu (Beijing) Biotechnology Co., Ltd., or Beijing Huafukang Biotechnology Co., Ltd. Mice were housed in a specific pathogen-free (SPF) room with a 12-h light / 12-h dark cycle and had free access to food and water.

[0054] Ethics Statement

[0055] All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) of the Fuwai Hospital, Chinese Academy of Medical Sciences. All animal experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

[0056] Establishment of a mouse myocardial infarction model

[0057] Adult mice with myocardial infarction were anesthetized with isoflurane and immobilized on a thermostatic plate. A 1.5 cm longitudinal incision was made approximately 1–2 mm lateral to the left sternal margin, and the chest wall muscles were bluntly dissected layer by layer. Rapid access to the thoracic cavity was performed through the third or fourth intercostal space. The intercostal space was opened with hemostats, and the heart was gently squeezed with the left hand in sync with the beating heart to eject it from the intercostal space. The left anterior descending coronary artery (LAD) was ligated with 7-0 Dacron suture 1–2 mm inferior to the left atrial appendage and 0.5 mm lateral to the pulmonary artery cone. Immediately, pallor of the ventricular myocardium below the ligature was observed. After ligation, the heart was gently returned to the chest cavity, the chest cavity was squeezed to expel air, and the skin incision was sutured. Quality control of the LAD occlusion was assessed by echocardiography one day after myocardial infarction.

[0058] Drug configuration

[0059] Apilimod (Sigma SML2974) was dissolved in PBS at a concentration of 0.5 mg / mL and injected into the tail vein.

[0060] Animal drug administration: The drug was injected into the tail vein before the establishment of the myocardial infarction model.

[0061] Detection index: EF (Ejection Fraction) value of the left ventricle of the mouse heart.

[0062] Detection method: Echo-cardiography (ultrasound detection)

[0063] Cardiac function in mice was assessed using a Vevo 3100 (FUJIFILM, Visualsonic, MS550D, or MS700) ultrasound system at baseline and 1, 7, and 14 days after myocardial infarction. M-mode echocardiographic images were obtained from each mouse in the parasternal short-axis position at the level of the mastoid muscle. At least five representative contraction cycles were selected, and the ejection fraction (EF) was calculated. All echocardiographic imaging and analysis were performed in a double-blind, randomized manner.

[0064] The model was considered successful when the EF value of the mouse cardiac ultrasound was between 10% and 30% at 1 dpi and was used to evaluate the efficacy of the drug. Figure 2 As shown in the results, the cardiac ultrasound EF (ejection fraction) values ​​of mice treated with Apilimod were significantly higher than those of the model control group at 7 and 14 dpi (*p<0.05, **p<0.01). The results indicate that Apilimod can significantly promote the recovery of cardiac function within 14 days after myocardial infarction.

[0065] The above description of the embodiments is intended to facilitate understanding and use of the invention by those skilled in the art. It will be apparent that those skilled in the art can readily make various modifications to these embodiments and apply the general principles described herein to other embodiments without requiring inventive effort. Therefore, the present invention is not limited to the above-described embodiments. Improvements and modifications made by those skilled in the art based on the disclosure of the present invention, without departing from the scope of the present invention, should be within the scope of protection of the present invention.

Claims

1. Use of a compound represented by formula (I) or a pharmaceutically acceptable form thereof in the preparation of a medicament for preventing or treating myocardial infarction:

2. The use according to claim 1, wherein the pharmaceutically acceptable form includes salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs.

3. The use according to claim 1, wherein the medicine further comprises a pharmaceutically acceptable carrier and / or excipient.

4. The use according to claim 3, wherein the pharmaceutically acceptable carrier and / or excipient comprises a solvent, a diluent, a dispersing aid, a suspending aid, a surfactant, an isotonic agent, a thickener or an emulsifier, a preservative, a solid binder, or a lubricant.

5. The use according to claim 1, wherein the dosage form of the drug includes tablets, capsules, granules, pills, drop pills, syrups, powders, suppositories, drops, emulsions, injections, solutions or suspensions.

6. The use according to claim 1, wherein the drug is administered in a single dose.

7. The use according to claim 1, wherein the drug is administered by intravenous injection.

8. The use according to claim 1, wherein the dosage of the drug is 2 mg / kg.

9. The use according to any one of claims 1 to 8, wherein the prevention or treatment of myocardial infarction comprises alleviating the effects of damage on myocardial cells and promoting recovery of cardiac function.

10. The use according to any one of claims 1 to 8, wherein the prevention or treatment of myocardial infarction comprises improving the systolic function of the left ventricle or improving the ejection fraction after myocardial infarction.

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