JP4888856B2 - Anti-angiogenic drug for ischemic disease - Google Patents

Description

  The present invention relates to a therapeutic agent for promoting angiogenesis for ischemic diseases.

  Specific indications according to the present invention include cerebral infarction, angina pectoris, myocardial infarction, and chronic obstructive arteriosclerosis.

  In the case of angiogenesis therapy using the therapeutic agent for promoting angiogenesis according to the present invention, oral administration, transvascular administration, etc. are possible in addition to direct administration to the affected area.

  The ischemic disease progresses chronically with arteriosclerosis as a basic pathological condition, leading to ischemia in the dominant region of the artery, and ultimately cell death and necrosis. The brain, heart, and extremities are known as arteries that progress in such a disease state and are clinically problematic.

  In the case of the brain, cerebral infarction, in the case of the heart, angina pectoris and myocardial infarction, and in the case of the extremities, chronic obstructive arteriosclerosis is the main disease name.

  Angiogenesis therapy is currently being tried for these diseases. A method is used in which bone marrow stem cells, peripheral blood stem cells, and hepatocyte growth factor are directly injected into the affected area. Bone marrow stem cells, peripheral blood stem cells, and hepatocyte growth factors are considered to have an effect of promoting angiogenesis.

1-Benzyl-4-[(5,6-dimethoxy-1-indanone) -2-yl] methylpiperidine is known as a therapeutic / preventive agent for various senile dementias such as Alzheimer-type senile dementia (described below). Patent Document 1).
Japanese Patent No. 2578475

  In the case of angiogenesis therapy using bone marrow stem cells and peripheral blood stem cells, in order to collect stem cells from bone marrow and peripheral blood vessels, special procedures, instruments, facilities, and time are required, and it is necessary to invade patients.

  In the case of angiogenesis therapy using bone marrow stem cells and peripheral blood stem cells, special procedures, instruments, facilities, and time are required for the work of separating and extracting stem cells from collected bone marrow fluid and peripheral blood.

  In the case of angiogenesis therapy using bone marrow stem cells, peripheral blood stem cells, and hepatocyte growth factor, special procedures, instruments, facilities, and time are required to inject these directly into the affected area.

  An object of the present invention is to provide a novel angiogenesis-promoting therapeutic agent for ischemic diseases.

  The present inventors have heretofore known donepezil (1-benzyl-4-[(5,6-dimethoxy-1-indanone) -2-yl] methylpiperidine), which is known as a therapeutic / preventive agent for various senile dementias. Was found to significantly promote angiogenesis and the present invention was completed.

That is, the therapeutic agent for promoting angiogenesis for an ischemic disease comprising angina pectoris, myocardial infarction, or chronic obstructive arteriosclerosis according to the present invention is 1-benzyl-4-[(5,6-dimethoxy-1-indanone). -2-yl] methylpiperidine and / or a salt thereof.

1-benzyl-4-[(5,6-dimethoxy-1-indanone) -2-yl] methylpiperidine: donepezil is an acetylcholinesterase inhibitor;
(1) Long plasma elimination half-life,
(2) Less peripheral side effects,
(3) High bioavailability and good brain migration,
These results were discovered as a result of an exploratory study as a compound that satisfies the above three items and has the effect of suppressing the progression of dementia symptoms in mild and moderate Alzheimer-type dementia.

  The mechanism of action of donepezil as a pro-angiogenic therapeutic agent is not necessarily clear, but donepezil acts on vascular endothelial cells, cardiomyocytes, and skeletal muscle cells to induce the expression of vascular endothelial growth factor and angiogenesis. It can be promoted.

  That is, donepezil acts as an inhibitor of acetylcholinesterase, and relieves dementia symptoms by indirectly increasing the acetylcholine concentration in the brain.

  The donepezil according to the present invention is considered to promote angiogenesis by activating the acetylcholine receptor on the endothelial cell membrane by increasing the acetylcholine concentration of the inhibitor.

  Hereinafter, embodiments of the present invention will be described in detail.

  The angiogenesis-promoting therapeutic agent for ischemic disease of the present invention comprises 1-benzyl-4-[(5,6-dimethoxy-1-indanone) -2-yl] methylpiperidine represented by the following structural formula (I) and / Or a pharmacologically acceptable salt thereof as an active ingredient.

  In the present invention, pharmacologically acceptable salts include, for example, inorganic acid salts such as hydrochloride, sulfate, hydrobromide, phosphate, formate, acetate, trifluoroacetate, maleate, Mention may be made of organic acid salts such as tartrate, methanesulfonate, benzenesulfonate and toluenesulfonate.

  Further, for example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt, N, N′-di In some cases, organic amine salts such as benzylethylenediamine salts, ammonium salts, and the like are formed.

  Donepezil may include geometric isomers, optical isomers, diastereomers, and the like, and all are included in the scope of the present invention.

  The method for producing donepezil is described in Patent Document 1 above.

  The angiogenesis-promoting therapeutic agent of the present invention can be orally administered to patients as tablets, pills, powders, powders, granules, syrups, solutions, suspensions, emulsions, capsules and the like. Further, it can be administered as an injection intravenously, intramuscularly, intradermally, subcutaneously, intraperitoneally, intraarterially, intrathecally, or the like. Furthermore, it may be administered into the rectum as a suppository, or can be implanted with a pellet. It can be administered as a spray, an inhalant, etc., or can be administered transdermally as an external preparation such as an ointment, cream, powder or liquid coating agent, patch.

  Among the above-mentioned, a preferable formulation form, administration form, and the like are appropriately selected by a doctor according to the patient's age, sex, constitution, symptoms, treatment timing, and the like.

Hereinafter, the present invention will be described more specifically with reference to examples.
[Example 1: Confirmation of angiogenesis-promoting effect in heart in rat myocardial ischemia model by coronary artery ligation]
<Method>
The left coronary artery of an 8 week old rat was ligated to obtain a chronic myocardial ischemia model. Two weeks after the operation, the rats were randomly divided into two groups, a group administered with donepezil and a group not administered with donepezil. The donepezil was dissolved in the drinking water of the group administered with donepezil. 4.8 mg.

  After 6 weeks of administration, we focused on von Willebrand Factor (vWF) and nicotinic acetylcholine receptor a7 subunit (a7-nAChR) molecules, which are thought to be related to angiogenesis, and applied immunohistochemistry to myocardial tissue. . A slice sample of 1 mm thickness was prepared from the extracted heart, and immediately immersed in 4% paraformaldehyde, the sample was macro-photographed. After fixing at 4 ° C. for 12 hours, it was embedded in paraffin to prepare 4 μm-thick continuous sections, which were subjected to the indirect fluorescent antibody method. After deparaffinization, the cells were treated in 0.01 M citrate buffer at 90 ° C. for 10 minutes and blocked with 10% goat normal serum for 1 hour. After treatment with an anti-vWF antibody (rabbit polyclonal) and an anti-a7-nAChR antibody (mouse monoclonal) at 4 ° C. for 12 hours, each secondary antibody, Alexa546-labeled goat anti-rabbit IgG and Alexa488-labeled goat anti-mouse IgG, was incubated at room temperature. Incubated for 4 hours and observed with a confocal laser microscope.

  FIG. 1 shows a heart specimen of a chronic myocardial ischemia model administered with donepezil. In FIG. 1, A and B are the results of macro imaging, and a marked neovascularization is observed in the ischemic region. C, D, and E are a transmission image, a vWF immune tissue image, and an a7nAChR immune tissue image of the same visual field, respectively, and show that the luminal structure in which the blood cell component is present is a blood vessel.

<result>
No vascular structure was confirmed in the chronic myocardial ischemia region of the left ventricle in the group not administered with donepezil. On the other hand, in the donepezil administration group, as shown in FIG. 1, a neovascular network was observed in the chronic myocardial ischemia region of the left ventricle. When the same specimen was observed by immunohistochemical technique, positive signals of vWF and a7-nAChR were observed along the vascular structure, and it was confirmed that blood cell components were clogged on the luminal side from the transmission image of the same visual field. It was.

<Conclusion>
From the above, it was found that donepezil has an angiogenic effect.

[Example 2: Confirmation of angiogenesis-promoting effect in the lower limbs in the chronic limb ischemia model by mouse femoral artery ligation]
<Method>
The left femoral artery of an 8-week-old mouse was ligated at both the groin and knee joint to form a chronic limb ischemia model. The right femoral artery was not ligated and used as a comparative control. After the operation, the mice are randomly divided into two groups, a group administered with donepezil and a group not administered with donepezil. I did it. The extent of angiogenesis was evaluated after 4 weeks of administration.

  FIG. 2A shows the macroscopic findings of the lower limb ischemia model in the donepezil-administered group and the donepezil non-administered group, FIG. 2B shows the histological findings by the HE staining, and FIG. 2C shows the VEGF immunohistochemical images.

<result>
As shown in FIG. 2, in the group not administered with donepezil, significant atrophy of skeletal muscles including the quadriceps muscle of the ischemic limb in which the femoral artery was ligated was observed.

  On the other hand, in the donepezil administration group, atrophy of skeletal muscles including the quadriceps femoris of the blood limbs was not observed, and many new blood vessels were observed in the histology. In the VEGF immunohistochemical image, more VEGF protein was stained in both the ischemic and non-ischemic side in the donepezil-administered group than in the non-administered group.

<Conclusion>
From the above, it was found that donepezil has an angiogenic effect.

[Example 3: Confirmation of angiogenic effect by human umbilical vein endothelial cells]
<Method>
Matrigel (registered trademark) (manufactured by BD Biosciences) and Dulbecco Modified Eagle's Medium were mixed at a volume ratio of 1: 1, and human umbilical vein endothelial cells (HUVEC) were cultured in the gelled medium to form luminal structures and The effect of donepezil on HUVEC VEGF production was evaluated.

  3A and 3B show the results 4 hours after the start of culture in the donepezil-treated and donepezil-treated, respectively.

<result>
From FIG. 3, remarkable growth of the luminal structure was recognized by donepezil treatment (0.1 to 1 micromolar concentration). Further, donepezil treatment promoted VEGF production in HUVEC.

<Conclusion>
From the above, it was found that donepezil has an angiogenic effect.

1 shows a heart specimen of a chronic myocardial ischemia model administered with donepezil in Example 1, FIG. 1A is a macrophotograph thereof, and FIG. 1B is a partially enlarged view of FIG. C is a transmission image, D is a vWF immune tissue image, and E is an a7nAChR immune tissue image. A, B and C in FIG. 2 are photographs showing the part where donepezil was administered and the lower limb gross findings when not administered, the tissue findings by HE staining, and the VEGF immunohistochemical images, respectively. FIG. 3 is a photograph showing the HUVEC lumen treated with donepezil and not treated with donepezil in Example 3.

Claims (1)

Angina pectoris, myocardial infarction, or chronic obstructive arteriosclerosis characterized by comprising 1-benzyl-4-[(5,6-dimethoxy-1-indanone) -2-yl] methylpiperidine and / or a salt thereof Angiogenesis-promoting therapeutic agent for ischemic disease consisting of symptoms .

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