JP6288625B2 - Heart failure inhibitor - Google Patents

Description

  The present invention relates to a medicament for suppressing heart fibrosis, ventricular wall thickening, cardiac hypertrophy, heart failure and aneurysm.

  If cardiac hypertrophy is prolonged, the risk of heart failure, sudden death, etc. increases. Therefore, in order to prevent or suppress heart failure, it is important to suppress cardiac hypertrophy. This cardiac hypertrophy is classified into a case where it occurs under physiological conditions due to exercise load and a case where it occurs under non-physiological conditions due to pathological stress reaction of the myocardium.

  On the other hand, reactive oxygen species (ROS) are known to be involved in signal transduction of growth in many cells including the cardiovascular system. In cardiomyocytes, it was proved that active oxygen mediates cardiomyocyte hypertrophy by angiotensin II and TNF-α, and active oxygen was shown to be involved in cardiomyocyte hypertrophy signaling as a second messenger (non-) Patent Document 1). The relationship between active oxygen and cardiac hypertrophy has also been reported at the cellular and animal levels, and active oxygen is rapidly produced after stimulation with the hypertrophic stimulants TNF-α, angiotensin II, endothelin-1 and phenylephrine. It is known that In addition, continuous subcutaneous administration of angiotensin II is widely known as a cardiac hypertrophy model, but it is also known that active oxygen is generated in the myocardium in this model.

  Cardiac hypertrophy, especially cardiac hypertrophy that occurs under non-physiological conditions due to the pathological stress response of the myocardium, is usually characterized by myocardial fibrosis and thickening of the ventricular wall. Sustained excessive stress on the myocardium due to long-term hypertension and abnormal stress on the myocardium after myocardial infarction often cause cardiac hypertrophy. It is known that when the cardiac hypertrophy persists, the heart fails to function as a pump, leading to heart failure.

  As drugs for such cardiac hypertrophy, statins, edaravone, β-blockers and the like are expected, but their effectiveness has not been confirmed yet.

  Aneurysms are the largest cause of subarachnoid hemorrhage, but the cause of the onset has not yet been clearly clarified. Although hypertension and arteriosclerosis are cited as risk factors for aneurysms, it is unclear whether they are the direct cause. As a means of treating an aneurysm, other than surgery, the progress of the aneurysm is only prevented with an antihypertensive drug.

Circulation 98: 794-799, 1998

  The subject of this invention is providing the pharmaceutical and foodstuff which suppress cardiac hypertrophy, heart failure, and aneurysm.

Therefore, as a result of various investigations to find a highly safe component that suppresses cardiac hypertrophy and aneurysm, the present inventors have found that luteolin or a derivative thereof, which is a kind of yellow pigment flavonoid contained in edible plants, is angiotensin in rats. In the II continuous administration model, it is found that it is useful as an inhibitor of heart failure prevention and aneurysm, significantly suppressing cardiac fibrosis, ventricular wall thickening, cardiac hypertrophy and aneurysm without affecting blood pressure. The present invention has been completed.
Furthermore, it has also been found that, among luteolin or its derivatives, luteolin-7-O-glucoside has high oral absorption and is useful as a medicine or food for oral consumption.

That is, the present invention provides a cardiac fibrosis inhibitor, a ventricular wall thickening inhibitor, a cardiac hypertrophy inhibitor, and a heart failure inhibitor containing luteolin or a derivative thereof as an active ingredient.
The present invention also provides an aneurysm inhibitor comprising luteolin or a derivative thereof as an active ingredient.
Furthermore, the present invention provides a composition for oral consumption containing luteolin-7-O-glucoside.

  Luteolin or a derivative thereof significantly suppresses cardiac fibrosis, ventricular wall thickening and cardiac hypertrophy, which are the causative symptoms of heart failure, and thus is useful as a pharmaceutical or food for preventing and / or suppressing heart failure. In addition, luteolin or a derivative thereof is useful as a medicine or food for preventing and / or suppressing an aneurysm because it significantly suppresses the aneurysm. Note that luteolin or a derivative thereof does not affect blood pressure in the angiotensin II continuous infusion model at doses exhibiting the above-described action, indicating that the above-described action is not an effect caused by lowering blood pressure. In addition, luteolin or a derivative thereof is particularly useful as a preventive agent for cardiac hypertrophy, heart failure, and aneurysm because it is safe and can be taken for a long time.

1 shows the effect of luteolin glycosides on blood pressure. It shows the effect of luteolin glycosides on ventricular wall thickening. It shows the effect of luteolin glycosides on heart and ventricular weight. It shows the effect of luteolin glycoside on the generation of active oxygen in the ventricular wall. The effect | action which acts on the fibrosis of the ventricular wall of a luteolin glycoside is shown. The effect | action which acts on the expression of the fibrosis marker gene of the ventricular tissue of a luteolin glycoside is shown. The effect of luteolin glycosides on cardiac hypertrophy and heart failure marker gene expression in ventricular tissues is shown. The effect | action which has on the blood pressure and a body weight with respect to the acute aneurysm model of a luteolin glycoside is shown. The effect | action (appearance) which acts on aneurysm formation of a luteolin glycoside is shown. The effect | action (diameter of aneurysm) which has on the aneurysm formation of a luteolin glycoside is shown. The effect | action (arterial wall elastic fiber tear suppression effect) which has an effect on the aneurysm formation of a luteolin glycoside is shown. The plasma concentrations of luteolin and its glycosides are shown.

  The active ingredient of the cardiac fibrosis inhibitor, ventricular wall thickening inhibitor, cardiac hypertrophy inhibitor, heart failure inhibitor, aneurysm inhibitor (hereinafter sometimes collectively referred to as a heart failure inhibitor) of the present invention is luteolin or Its derivatives. Luteolin or a derivative thereof has the following formula (1)

(Wherein R ^{1 to} R ⁴ each independently represents a hydrogen atom, an alkyl group, a sulfo group or a sugar residue)

It is a kind of yellow pigment flavonoid represented by. Of these, luteolin or a glycoside thereof is particularly preferable.

Examples of the alkyl group represented by R ^{1 to} R ⁴ include an alkyl group having 1 to 4 carbon atoms, among which a methyl group, an ethyl group, an n-propyl group, and an isopropyl group are preferable, and a methyl group is more preferable. A sulfo group is a group represented by —SO ₃ H. Examples of sugars constituting the sugar residues represented by R ^{1 to} R ⁴ include monosaccharides such as glucose, galactose, xylose, mannose, and glucuronic acid; disaccharides such as apiosylglucoside, maltose, cellobiose, and gentibiose; A mono- or disaccharide acetylated form, malonylated form, etc. are mentioned. Of these sugar residues, residues derived from monosaccharides or disaccharides are more preferred.

Of these luteolins or glycosides thereof, luteolin (R ^{1 to} R ⁴ = H) and luteolin-7-O-glycoside (R ¹ = monosaccharide or disaccharide, R ^{2 to} R ⁴ = H) are preferred. , Luteolin and luteolin-7-O-glucoside (R ¹ = glucose residue, R ^{2 to} R ⁴ = H) are more preferred, and luteolin-7-O-glucoside is particularly preferred from the viewpoint of oral absorption.

  These luteolins or their derivatives may be contained in foods such as eggplant, sesame, perilla, asteraceae, seriaceae, cruciferous, gramineous, legumes, rose, honeysuckle, camellia, etc. Are known. Examples of solanaceous plants include plants belonging to the genus Capsicum, and among them, they are contained in large amounts in capsicum, peppers, paprika and the like. Sesame plants include sesame plants, which are abundant in sesame. In addition, as a plant of the family Lamiaceae, there is a genus of perilla such as egoma and perilla, mint genus such as mint and peppermint, rosmarinus genus such as rosemary, genus Rosaliaus such as thyme, genus genus such as oregano, and genus Akigiri such as sage. Plant. Examples of the Asteraceae plants include plants belonging to the genus Shungyoku such as Shungyiku, the genus Achinonokeshi such as lettuce, the deer genus such as chamomile, and the genus Dandelion such as dandelion. Examples of the plants of the celery family include plants of the genus Dutch honey, such as celery, the genus Dutch genus, such as parsley, the genus Sisido such as Ashitaba, and the carrot genus of carrot sugar. Examples of the cruciferous plants include plants of the genus Brassica such as broccoli and cabbage. Examples of the grass family plant include sugarcane. Examples of leguminous plants include peanuts and high bosses. Examples of rose family plants include apple apples, examples of honeysuckle family include honeysuckle, and examples of camellia family include tea tree. The extraction part is preferably an edible part of these plants.

To extract luteolin or its derivatives from these edible plants, for example, hot water, alcohols such as ethanol, esters such as ethyl acetate, ethers such as diethyl ether and dioxane, halogenated hydrocarbons such as chloroform and dichloromethane, It is preferable to use water or an organic solvent such as ketones such as acetone. Of these, extraction with water or ethanol is particularly preferred. The extraction is preferably performed at a temperature of 0 ° C. to the boiling point of the solvent for 1 hour to 72 hours. The extract can be further purified by column chromatography or the like. Moreover, a commercial item may be used for luteolin or its derivative (s).
Also, luteolin-7-O-glucoside (7-O-β-D-glucosyl luteolin) can be obtained by acid hydrolysis of luteolin-7-O-apiosyl (1-2) -glucoside (JP 2008-2008). -20175).

  In the heart failure inhibitor of the present invention, luteolin or a derivative thereof may be blended, or a plant extract containing luteolin or a derivative thereof may be blended.

  Luteolin or a derivative thereof has excellent cardiac fibrosis inhibitory effect and ventricular wall thickening inhibition without affecting blood pressure in hypertrophy, heart failure model, and aneurysm model by continuous infusion of angiotensin II, as shown in Examples below. Action, cardiac hypertrophy inhibiting action, heart failure inhibiting action and aneurysm inhibiting action. Therefore, luteolin or a derivative thereof is particularly useful as a cardiac hypertrophy inhibitor, heart failure inhibitor, or aneurysm inhibitor.

  The heart failure inhibitor of the present invention can be used not only as a pharmaceutical product but also as a quasi-drug, a food for specified health use, a functional food (oral supplement, health food, nutritional supplement, hospital food, medical food, etc.). .

  The medicament of the present invention can be made into various administration forms by blending luteolin, derivatives thereof or plant extracts containing these with a pharmaceutical carrier as necessary, and includes, for example, oral preparations, Examples include injections, suppositories, ointments, patches, and the like, but oral agents are preferred. Examples of oral preparations include tablets, granules, fine granules, powders, capsules, syrups and the like.

  As the pharmaceutical carrier, various organic or inorganic carrier substances commonly used as pharmaceutical materials are used. Excipients, binders, disintegrants, lubricants, colorants in solid preparations; solvents, dissolution aids, suspensions in liquid preparations. It is blended as a turbidity agent, tonicity agent, buffering agent, soothing agent and the like. Moreover, formulation additives such as preservatives, antioxidants, colorants, sweeteners, stabilizers and the like can be used as necessary.

  When preparing an oral solid preparation, add luteolin, its derivatives or plant extracts containing them with excipients, if necessary, binders, disintegrants, lubricants, coloring agents, flavoring and flavoring agents, etc. After the addition, tablets, coated tablets, granules, powders, capsules and the like can be produced by conventional methods.

  When preparing a liquid preparation for oral use, add a flavoring agent, buffering agent, stabilizer, flavoring agent, etc. to luteolin, its derivatives or plant extracts containing them, etc., and oral liquids, syrups, elixirs, etc. Can be manufactured.

  When preparing injections, add pH adjusters, buffers, stabilizers, isotonic agents, local anesthetics, etc. to luteolin, its derivatives or plant extracts containing them, and then apply subcutaneously, Internal and intravenous injections can be produced.

  When preparing a suppository, luteolin, a derivative thereof or a plant extract containing them is added to a pharmaceutical carrier known in the art, for example, polyethylene glycol, lanolin, cocoa butter, fatty acid triglyceride and the like, if necessary, Tween 80. After adding a surfactant such as (registered trademark), it can be produced by a conventional method.

  When preparing an ointment, bases, stabilizers, wetting agents, preservatives and the like that are usually used in luteolin, its derivatives or plant extracts containing them are blended as necessary, and mixed and formulated by conventional methods. It becomes.

  When preparing a patch, the ointment, cream, gel, paste or the like may be applied to a normal support by a conventional method.

The amount of luteolin or a derivative thereof to be incorporated in the above-mentioned pharmaceutical or functional food is not constant depending on the symptoms of the patient to which this is to be applied, or its dosage form, but generally an oral preparation per dosage unit form Is about 0.05 to 1000 mg, about 0.01 to 500 mg for injections, and about 1 to 1000 mg for suppositories.
In addition, the daily dose of the pharmaceutical having the above dosage form varies depending on the patient's symptoms, body weight, age, sex, etc., and cannot be determined unconditionally. It is about 5000 mg, preferably 0.1 to 1000 mg, and is preferably administered once a day or divided into 2 to 3 times a day.

  Among luteolin and its glycosides, luteolin-7-O-glucoside is significantly faster and more excellent in oral absorption than luteolin and luteolin-7-O-apiosylglucoside. Therefore, the composition for oral consumption containing luteolin-7-O-glucoside is particularly useful as a composition for medicines and foods.

  EXAMPLES Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

Example 1
Effects of luteolin glycosides (using luteolin-7-O-glucoside) on cardiac fibrosis / hypertrophy in an oxidative stress-enhanced state by continuously injecting angiotensin II into 8-week-old male SD rats investigated. In the control group, rats were normally bred. In angiotensin II group (Ang II group), angiotensin II continuous infusion pump was implanted subcutaneously into rats and infused continuously for 7 days. Luteolin glycoside coadministration group (Ang II + LMG group) was loaded with luteolin glycoside mixed diet (containing 3.6 g luteolin monoglycoside (LMG) per 10 kg of feed) for 3 weeks and then angiotensin II continuous infusion pump subcutaneously Implanted and infused continuously for 7 days.
On the seventh day, blood pressure was measured, and after evaluating the ventricular wall thickness by echocardiography, the heart was removed and the weight of the heart and ventricle was measured. Reactive oxygen was evaluated by staining with hydroethidine, and the degree of fibrosis was evaluated by Masson trichrome staining. Gene expression of fibrosis markers and heart failure markers in ventricular tissues was evaluated by real-time PCR.

  As a result, as shown in FIG. 1, the luteolin glycoside did not decrease the blood pressure increased by continuous administration of angiotensin II. In addition, as shown in FIG. 2, ventricular wall thickening due to angiotensin II administration (Ang II) was significantly suppressed by administration of luteolin glycoside. As shown in FIG. 3, the same effect was observed in the evaluation of heart weight / ventricular weight. As a result, it can be seen that luteolin glycosides do not affect blood pressure, effectively act on the heart, and suppress cardiac hypertrophy.

  Moreover, as shown in FIG. 4, the luteolin glycoside significantly suppressed the generation of active oxygen in the ventricular wall by angiotensin II. Furthermore, as shown in FIG. 5, the luteolin glycoside markedly suppressed ventricular wall fibrosis by angiotensin II.

  RNA was extracted from ventricular tissue, and various gene expression levels were evaluated by real-time PCR. As shown in FIG. 6, TGF-β1 (transforming growth factor) and CTGF (connective tissue growth factor) gene expression reflecting fibrosis is an internal standard GAPDH expression, and after expression is activated by angiotensin II and luteolin distribution. Suppression of expression due to the combined use of saccharides was observed, and the results were consistent with the degree of fibrosis (FIG. 5). As shown in FIG. 7, gene expression of ANP (atrial natriuretic peptide) and BNP (brain natriuretic peptide) reflecting cardiac hypertrophy / heart failure is an internal standard GAPDH expression, and after expression is activated by angiotensin II, luteolin glycoside Suppression of expression due to combined use with the body was observed, and the results were consistent with the degree of cardiac hypertrophy (FIGS. 2 and 3).

Example 2
A 12-week-old ApoE gene-deficient female mouse (ApoE − / −) (weight approximately 25 g) is continuously infused with angiotensin II (from 16 to 20 weeks) to prepare an acute aneurysm model, and luteolin glycoside (luteolin- 7-O-glucoside was used). In the control group (n = 5), rats were normally bred (continuous infusion of physiological saline). In angiotensin II group (Ang II tn-rmol group), angiotensin II continuous infusion pump was implanted subcutaneously into rats and infused continuously for 4 weeks. Luteolin glycoside coadministration group (Ang II + luteolin group) is an angiotensin II continuous infusion pump subcutaneously after loading with a luteolin glycoside compounded diet (containing 0.055% luteolin-7-O-glucoside per bait) for 3 weeks Were implanted for 4 weeks.

  At 20 weeks of age, blood pressure and body weight were measured, and then the heart, kidney and aorta were removed, and the occurrence of aneurysm was observed.

  As a result, as shown in FIG. 8, there was no significant change in blood pressure and body weight at this dose of luteolin glycoside. As shown in FIG. 9, the luteolin glycoside administration group clearly suppressed the formation of an aneurysm. As shown in FIG. 10, in the luteolin glycoside administration group, the diameter of the aneurysm was small. In addition, as shown in FIG. 11, the luteolin glycoside administration group suppressed arterial wall elastic fiber rupture.

Example 3
Luteolin (51 mg / kg, PO), luteolin-7-O-glucoside (93 mg / kg, PO) and luteolin-7-O-apiosylglucoside (175 mg / kg, PO) were administered to rats (8 weeks old, male SD system). Rats were orally administered, and blood was collected from the tail vein immediately before administration, 0.5 hours, 1 hour, 2 hours and 8 hours later, and the concentration of luteolin in plasma was measured. The dose of each luteolin and its glycoside was the same as the amount of luteolin that is an aglycone.
The result is shown in FIG. As is clear from the results of FIG. 12, luteolin-7-O-glucoside showed significantly higher oral absorption than luteolin and luteolin-7-O-apiosylglucoside.

Claims (2)

Cardiac fibrosis-suppressing agent comprising as an active ingredient Le Teorin -7-O-glucoside. Le Teorin -7-O-glucoside cardiac fibrosis-suppressing food composition comprising, as an active ingredient.