WO2008015950A1 - Acat inhibitor - Google Patents

Abstract

An acyl-CoA cholesterol acyltransferase inhibitor comprising at least one member selected from the group consisting of 3'-acetoxy-4'-tigloyloxy-3'-4'-dihydroseselin, a derivative thereof and a pharmaceutically acceptable salt of the compound or the derivative as an active ingredient. The inhibitor or a pharmaceutical, food or animal feed comprising the inhibitor is effective for hyperlipemia, arterial sclerosis or a disease associated with these diseases. Particularly, the food is useful as a functional food which has an inhibitory activity on acyl-CoA cholesterol acyltransferase and is effective for the maintenance of homeostasis in vivo.

Description

 Specification

 ACAT inhibitor

 Technical field

 [0001] The present invention relates to a component derived from a natural product having an action of inhibiting an acyl-CoA cholesterol acyltransferase and useful for the treatment or prevention of arteriosclerosis, hyperlipidemia and the like. Background art

 [0002] In recent years, arteriosclerosis, hyperlipidemia, hypertension, diabetes mellitus and leprosy have been accompanied by changes in dietary intake of high-calorie and high-cholesterol diets and an increase in the elderly with increased life expectancy. , Tsuta! /, The increase in the number of lifestyle-related diseases is rapidly increasing, and it has become a big social problem! /,

[0003] In early lesions of arteriosclerosis, spotted or linear fat deposits called fat streaks are seen. This change is mainly due to the accumulation of foamed macrophages on the vascular endothelium. Macrophage foaming occurs when macrophages take up denatured LDL to produce free cholesterol, which is esterified by acyl CoA cholesterol acyltransferase, which accumulates. Such early foaming cell lesions progress to complex lesions involving foaming of vascular smooth muscle cells. The fatty streak eventually becomes a fibrous hard spot and protrudes into the blood vessel wall, and as the lesion progresses further, calcification and thrombus attachment accompany the narrowing of the blood vessel cavity, and the hard spot breaks down to cause thrombotic occlusion. Come on. It is also known that tablets are easy to break! /, And hard spots are rich in lipid components such as cholesterol esters! /. Therefore, inhibiting the activity of acyl-CoA cholesterol-acyltransferase leads to stabilization and regression of arteriosclerotic lesions, leading to the onset and recurrence of acute coronary syndromes based on arteriosclerosis and hyperlipidemia. Be expected. Inhibition of the acyl CoA cholesterol facilyltransferase activity reduces the synthesis of cholesteryl ester that constitutes VLDL in the liver, resulting in a decrease in VLDL synthesis, and inhibition of cholesterol ester binding in the small intestine. Is known to decrease, blood cholesterol and blood neutral fat can be expected to decrease. [0004] The button bow is a perennial belonging to the genus Waraboufu, which has the scientific name Peucedanum japonicum. Put a lot of flowers. Its physiological activity is said to be effective in cold, fatigue recovery, and nourishing tonic in its origin, Okisumari, and has been useful for a long time. In recent studies, button bow fu has a cancer suppressive effect (eg, Non-patent Document 1), a disaccharide-degrading enzyme inhibitory action (eg, Patent Document 1), an antioxidant action (eg, Patent Document 2, Non-patent Document 2) It is also known that there are cell activation effects (for example, Patent Document 2) and melanin production suppression effects (for example, Patent Document 2).

 [0005] As a characteristic chemical component contained in button bow fu, a plurality of types of coumarin derivatives are known, and their tumor promoter suppression action and the like have been studied (for example, Non-Patent Document 3).

 Patent Document 1: Japanese Patent Laid-Open No. 2003-26694

 Patent Document 2: JP 2004-26697 A

 Non-patent literature 1: T. Morioka and 8 others, Cancer Letters, 2004, Vol. 205, pl 33-141

 Non-Patent Document 2: Hisamoto et al., J. Agric. Food Chem., 2004, Vol. 5 2, p445 -450

 Non-Patent Document 3: Fan · 5 others, Journal of Japanese Botany, 2000, Vol. 7 5, No. 4, p257-261

 Summary of invention

 [0006] To outline the present invention, the first invention of the present invention consists of 3'-acetoxy 4'-tigroinoloxy 3 ', 4' dinoid roseserine, its derivatives and pharmaceutically acceptable salts thereof. The present invention relates to an acyl CoA cholesterol acyltransferase inhibitor characterized by containing at least one selected from the group as an active ingredient.

 [0007] A second invention of the present invention relates to a pharmaceutical comprising the acyl CoA cholesterol acyltransferase inhibitor of the first invention of the present invention.

[0008] A third invention of the present invention relates to a food or beverage comprising the acyl CoA cholesterol acyltransferase inhibitor of the first invention of the present invention. Detailed Description of the Invention

 [0009] The present invention relates to a pharmaceutical, food or feed that uses a substance having an inhibitory action on an Asinole CoA cholesterol acyltransferase suitable as a food material or a pharmaceutical material that can be safely and easily ingested.

 [0010] According to the present invention, a group consisting of 3'-acetoxy 4'-tigroyloxy 3, 4, 4'-dihydroserine, a derivative thereof, and a pharmaceutically acceptable salt thereof, which are components derived from natural products Provided are an acyl CoA cholesterol transferase inhibitor containing at least one selected from the above as an active ingredient, and a pharmaceutical, food or feed containing the inhibitor. The inhibitor, medicine, food or feed is useful for hyperlipidemia, arteriosclerosis, and diseases caused by them, and among them, the food of the present invention has an inhibitory action on acyl-CoA cholesterol acyltransferase. It is useful as a functional food useful for maintaining homeostasis.

 [0011] Acyl CoA cholesterol acyltransferase (also referred to as Facil CoA: cholesterol O-acyltransferase; hereinafter sometimes abbreviated as ACAT) transfers long chain fatty acids from isacyl CoA to cholesterol, It is an enzyme that catalyzes the synthesis of cholesterol esters. The ACAT inhibitory action can be easily measured using an assay system as described in Example 1 described later. That is, by mixing a test substance and an enzyme source containing ACAT and evaluating the transfer of the Oleoyl group from radiolabeled Oleoyl-CoA to cholesterol, the ACAT inhibitory effect can be easily measured.

 [0012] In the following formula (I), 3'-acetoxy 4'-tigroyloxy 3 ', 4'-dihydroserine

 A structural formula (hereinafter, sometimes referred to as a compound of the present invention) is shown.

 [0013] [Chemical 1]

[0014] 当該化合物はボタンボウフゥゃ前胡（Peucedanum praeruptorum)に含まれる 物質であり、本発明において ACAT阻害作用を有することが初めて明らかにされた。 すなわち、本発明における有効成分としては、 3 '—ァセトキシ— 4 '—チグロイルォキ シー 3 ' , 4 ' ジノヽイドロセセリン、その誘導体及び/又は薬学的に許容されるそれら の塩 (以下、これら全てを本発明の有効成分と称することがある）が使用される。

[0014] This compound is a substance contained in Peucedanum praeruptorum and was first demonstrated to have an ACAT inhibitory action in the present invention. That is, as the active ingredient in the present invention, 3′-acetoxy-4′-tigloyloxy 3 ′, 4 ′ dinoid roseserine, its derivatives and / or pharmaceutically acceptable salts thereof (hereinafter, all of these Which may be referred to as the active ingredients of the invention).

 [0015] The method for producing the compound of the present invention is not particularly limited, but button boo cucumber (Peucedanum praeruptorum) can be used as a raw material. For example, as described in Preparation Example 1, various methods can be used from button boufu ethanol extract. The power that can be obtained through chromatography. In addition, when using button bow as a raw material, there is no particular limitation. The ability to use fruits, seeds, seed coats, flowers, leaves, stems, roots, rhizomes and / or whole plants as they are. / Or stems can be used. In addition, when the compound of the present invention is synthesized, it can also be obtained by combining known methods.

 [0016] In the present specification, 3'-acetoxy 4'-tigroyloxy 3 ', 4'-dihydroserine is any isomer such as any optical isomer, ketoeenol tautomer, geometric isomer, etc. Even so, as long as it has an ACAT inhibitory action, it can be used in the present invention with a force S.

 [0017] In the present specification, as the derivative, for example, various substituents are bonded to the skeleton of 3'-acetoxy-4 'tiglyloyloxy-3', 4'-dino or idrothelin as described above to exhibit a desired effect. There is no particular limitation as long as it is possible. Examples of the substituent include aliphatic groups (straight chain aliphatic groups such as methyl, ethyl, and n-propyl groups, and branched aliphatic groups such as isopropyl, isobutyl, prenyl, and geranyl groups). , Aromatic group (phenyl group, naphthyl group, biphenyl group, pyrrolyl group, indolyl group, etc.), aromatic aliphatic group (benzyl group, phenethyl group, etc.), hydroxyl group, carboxyl group, sulfate group, phosphate group, thiol Groups, amino groups, nitro groups, alkoxy groups (such as methoxy groups), acyloxy groups (such as acetyl groups), and halogens (such as chlorine, bromine and fluorine). In addition, for example, a substance obtained by metabolizing the compound of the present invention to a mammal is also included in the derivative of the present invention.

[0018] As the salt of 3'-acetoxy 4'-tigroyloxy 3 ', 4'-dihydrate mouth ceserine or a derivative thereof used in the present invention, a pharmaceutically acceptable salt is preferable. In the present invention Examples of the salt used include alkali metal salts, alkaline earth metal salts, and salts with organic bases. The pharmaceutically acceptable salt used in the present invention means a salt of a compound that is substantially non-toxic to living organisms and has an ACAT inhibitory action. Examples of the salt include sodium, potassium, calcium, magnesium, ammonium, or protonated benzathine (N, N, benzylene ethylenediamine), choline, ethanolamine, diethanolamine, Examples thereof include salts of ethylenediamine, megramin (N-methyldulamine), venetamine (N-benzylphenethylamine), piperazine or tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol).

 [0019] Derivatives of the compounds of the present invention can be prepared by methods known in the art. In addition, a salt of the compound or derivative of the present invention can be easily prepared by a person skilled in the art by any method known in the art.

 [0020] The active ingredient of the present invention is a compound contained in button bow fowl that has been edible since ancient times, and a synthetic compound having an ACAT inhibitory action known so far (for example, FR179254 described later). As will be described later, there is no particular toxicity, and there is no risk of side effects. Therefore, it is possible to safely and appropriately treat or prevent diseases. Therefore, an ACAT inhibitor comprising the active ingredient (hereinafter sometimes referred to as the ACAT inhibitor of the present invention) is effective in the treatment or prevention of a disease that requires an ACAT inhibitory action, and is described below as a pharmaceutical or food product. Alternatively, it is useful as a feed material. The ACAT inhibitor of the present invention is also used for screening drugs for diseases that require ACAT inhibitory action for treatment or prevention, such as arteriosclerosis and hyperlipidemia, and diseases caused by these factors. Useful. Furthermore, the ACAT inhibitor is useful for studying the mechanism of cholesterol ester production and the mechanism of the development of the above-mentioned diseases such as hyperlipidemia and arteriosclerosis.

[0021] The ACAT inhibitor of the present invention is not particularly limited as long as it contains the above-mentioned active ingredient of the present invention. For example, the active ingredient of the present invention itself, or a composition containing the active ingredient of the present invention. It may be a thing. That is, the content of the active ingredient in the ACAT inhibitor is determined in consideration of the intended use of the ACAT inhibitor. The amount is not particularly limited as long as the expression of the fruit can be obtained. The content of the active ingredient in the ACAT inhibitor of the present invention is usually about !! to 100% by weight. Further, the amount of the ACAT inhibitor used is not particularly limited as long as the desired effect of the present invention can be obtained. What is necessary is just to use it in the quantity which can administer an active ingredient within the dose range of the active ingredient in the pharmaceutical of this invention. The administration method is not particularly limited, and may be appropriately set in the same manner as the medicine described below. Further, the form of use is not particularly limited, and it can be produced in an appropriate form according to the intended use.

 [0022] The present invention also provides a medicament containing the ACAT inhibitor of the present invention (hereinafter referred to as "medicament of the present invention S"). The medicament of the present invention is useful for diseases requiring ACAT inhibitory action for treatment or prevention, and the diseases are not particularly limited. For example, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, Metabolic syndrome, multiple risk factor syndrome, arteriosclerosis, ischemic heart disease, acute myocardial infarction, unstable angina, sudden ischemic death, cerebrovascular disorder, chronic obstructive arteriosclerosis, myocardial infarction, angina , Cerebral infarction, subarachnoid hemorrhage, obesity or diseases caused by these (for example, O 'Rourke et al., J. Biol. Chem., 2002, 277 (45), 42557-42562, (See Ohishi et al., Biol. Pharm. Bull., 2003, 26 (8), 1125-1128, and Ohishi et al., Chem. Pharm. Bull., 2001, 49 (7), 830-839).

 [0023] Examples of the medicament of the present invention include those prepared by combining the above-mentioned effective component used as the ACAT inhibitor according to the present invention with a known pharmaceutical carrier. Moreover, as the medicament of the present invention, the active ingredient of the present invention can be blended with other ingredients that can be used for the same use as the active ingredient as described above.

[0024] The production of the medicament of the present invention is usually carried out by blending the above active ingredient with a pharmaceutically acceptable liquid or solid carrier, and if necessary, a solvent, a dispersant, an emulsifier, a buffer, Agents, excipients, binders, disintegrants, lubricants, etc. are added to form solids such as tablets, granules, powders, powders, capsules, etc., and usually liquids such as liquids, suspensions, and emulsions. be able to. In addition, dry products that can be made liquid by adding an appropriate carrier before use, and other externally used IJs are also included. [0025] The pharmaceutical carrier can be selected depending on the administration form and formulation of the pharmaceutical. In the case of an oral preparation composed of a solid composition, it can be a tablet, pill, capsule, powder, fine granule, condyle granule, etc., for example, starch, lactose, sucrose, mannitol, carboxymethy Pharmaceutical carriers such as cellulose, corn starch and inorganic salts are used. In preparation of oral preparations, binders, disintegrants, surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents, fragrances and the like can be further added. For example, in the case of tablets or pills, it may be coated with a sugar coating such as sucrose, gelatin or hydroxypropylcellulose, or a film of a gastric or enteric substance, if desired. In the case of an oral preparation comprising a liquid composition, it can be a pharmaceutically acceptable emulsion, solution, suspension, syrup, etc. For example, purified water, ethanol or the like is used as a carrier. The Further, if desired, adjuvants such as wetting agents and suspending agents, sweetening agents, flavoring agents, preservatives and the like may be added.

 [0026] On the other hand, in the case of a parenteral preparation, distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil according to a conventional method, using the above active ingredient of the present invention as a diluent. It can be prepared by dissolving or suspending in corn oil, propylene glycol, polyethylene glycol, etc., and adding bactericides, stabilizers, tonicity agents, soothing agents, etc. as necessary. In addition, a solid composition can be produced and used by dissolving in sterile water or a sterilized solvent for injection before use.

 [0027] The external preparation includes solid, semi-solid or liquid preparations for transdermal administration or transmucosal (intraoral or intranasal) administration. Also included are suppositories and the like. For example, emulsions such as emulsions and lotions, liquid preparations such as external tinctures, liquids for transmucosal administration, ointments such as oily ointments and hydrophilic ointments, transdermal such as films, tapes, and poultices It may be a patch for administration or transmucosal administration.

[0028] The medicaments in the various preparation forms as described above can be appropriately produced by conventional methods using known pharmaceutical carriers and the like. In addition, the content of the active ingredient in force and medicinal medicine is particularly limited as long as the active ingredient can be administered within the dosage range described below, taking into consideration its administration form, administration method and the like. It is not something. The content of the active ingredient in the medicament of the present invention is usually about 1 to 100% by weight. [0029] The medicament of the present invention is administered by an appropriate administration method according to the preparation form. The administration method is not particularly limited, and for example, it can be administered by internal use, external use or injection. When the pharmaceutical agent of the present invention is administered by injection, it can be administered, for example, intravenously, intramuscularly, subcutaneously, intradermally, and when administered externally, for example, as an external preparation such as a suppository, its suitable administration It can be administered by any method.

 [0030] The dosage of the medicament of the present invention is appropriately set according to the formulation form, administration method, purpose of use, and age, weight, and symptoms of the patient to whom the medicament is administered and is not constant. In general, the dosage of the active ingredient contained in the preparation is preferably 0.1 μg to 5 g / kg body weight, more preferably 1 μg to 2 g / kg body weight, more preferably 10 per day for an adult. 〃 g ~ lg / kg body weight. Of course, since the dose varies depending on various conditions, an amount smaller than the above dose may be sufficient or may be necessary beyond the range. Administration may be carried out once or within several days within a desired dose range. The administration period is also arbitrary. In addition, the medicament of the present invention can be orally administered as it is orally added to any food.

 [0031] As another aspect of the present invention, a method for treating a disease using the medicament is provided. The method for treating the disease comprises treating the above-mentioned medicament containing the active ingredient of the present invention with hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, metabolic syndrome, multiple risk factor syndrome, arteriosclerosis, Bloody heart disease, acute myocardial infarction, unstable angina, ischemic sudden death, cerebrovascular disorder, chronic obstructive arteriosclerosis, myocardial infarction, angina, cerebral infarction, subarachnoid hemorrhage, obesity or It is characterized by administration to patients with diseases that occur as causative factors. The conditions for administration of the drug can be implemented in accordance with the aforementioned disclosure of drug administration.

 [0032] Further, as another aspect of the present invention, there is provided use of 3'-acetoxy 4 'tigloroxy-3', 4'-dino or idroserine for producing the medicament. 3′-Acetoxy-4′-tigroyloxy-3 ′ and 4′-dihydroserine include the aforementioned derivatives and pharmaceutically acceptable salts thereof.

[0033] The present invention provides an ACAT-inhibiting food or feed comprising the ACAT inhibitor of the present invention (sometimes referred to herein as the food or feed of the present invention). . The food or feed of the present invention has an ACAT inhibitory action of the active ingredient of the present invention. Very useful for prevention. That is, the food of the present invention is extremely useful as a functional food (specific health food, etc.) with the purpose of preventing, ameliorating or treating the above-mentioned diseases, and is concerned about blood cholesterol level. It is a very useful food for those who are concerned about neutral fat and those who are concerned about body fat. In the specification of the present application, food includes drinks.

 [0034] In the food or feed of the present invention, "containing" means containing, adding and / or diluting. Here, “containing” means that the active ingredient used in the present invention is contained in food or feed, and “addition” means that the active ingredient used in the present invention is added to the raw material of food or feed. The “dilution” refers to the addition of food or feed ingredients to the active ingredient used in the present invention.

 [0035] The food of the present invention is not particularly limited. For example, a processed grain product (processed flour product, processed starch product, premix processed product, rice cake) containing the active ingredient according to the present invention is included. ), Macaroni, breads, bean paste, buckwheat, rice cake, rice noodles, harsame, packaging rice cake etc.) Miso, natto, etc.), processed meat products (ham, bacon, press ham, sausage, etc.), marine products (frozen surimi, force, maboko, chikuwa, champagne, fried fish cake)

, Tsumire, Line, Fish ham, Sausage, Bonito, Processed egg products, Canned fish, Boiled Tsukuda, etc.), Dairy products (Ingredient milk, cream, yogurt, butter, cheese, condensed milk, powdered milk, ice cream, etc.), vegetables 'Processed fruit products (pastes, jams, pickles, fruit drinks, vegetable drinks, mixed drinks, etc.), confectionery chocolate, biscuits, confectionery breads, cakes, candy sweets, rice confectionery etc.), alcoholic drinks (Japanese sake) , Chinese liquor, wine, whiskey, shochu, wow, brandy, gin, rum, beer, soft drink, fruit liquor, liqueur, etc., beverages (green tea, tea, oolong tea, coffee, soft drink, lactic acid) Beverages, etc.), seasonings (soy sauce, sauce, vinegar, mirin, etc.), canned food, bottled food, bagged food (beef rice, kettle rice, red rice, curry, other Such as seeds cooked food), semi-dry or concentrated foods (lever paste, other spreads, buckwheat 'udon soup, such as concentrated soups), dry food (instant 麵類 , Instant Curry, Instant Coffee, Powdered Juice, Powdered Soup, Instant Miso Soup, Cooked Food, Cooked Beverage, Cooked Soup, etc., Frozen Food (Sukiyaki, Chawanmushi, Unagi Kabayaki, Hamburg Steak, Syumai, Dumplings, Various Sticks , Fruit vegetables, etc.), solid foods, liquid foods (soups, etc.), spices and other agricultural products such as processed forest products, processed livestock products, processed fishery products.

[0036] In the food of the present invention, if the active ingredient is contained singly or plurally, contained, added and / or diluted, and the content thereof corresponds to a necessary amount for expressing the ACAT inhibitory action, In particular, powders, tablets, granules, capsules and the like that can be taken orally are also included. In addition, the food of the present invention includes foods obtained by mixing the above-mentioned active ingredients of the present invention as they are or appropriately mixed with appropriate emulsifiers and excipients. These foods can be eaten as drinks as they are or mixed with water.

 [0037] In addition, the food of the present invention can be mixed with the active ingredient of the present invention and the juice of various plants (vegetables, fruits, etc.) to make a health drink. For example, by diluting the active ingredient of the present invention with water, button bouffwa, ij ij old moon Dalefruit, Kiwi, Spinach, Radish, Radish, Chinese cabbage, Cabbage, Salad, Lettuce, Yura, Okra, Pepper, Cucumber, Green beans, Edamada, Endo, Corn, Herb, Summer tangerine, Sweet mandarin It can be mixed with squeezed juice in summer, milk, soy milk, etc. to make a health drink having ACAT inhibitory action.

 [0038] Further, as an aspect of the food of the present invention, 3'-acetoxyl-4'-tigroyloxy-3 ', 4'-dihydroserine is used as an active ingredient (participating ingredient), and treatment of plants containing the same Also included is an embodiment in which a product such as an extract or pulverized product of button bow fu, squeeze juice, or the like is used as it is as a food for which the effect of the active ingredient of the present invention is expected, for example, a food for specified health use.

[0039] The content of the active ingredient in the food of the present invention is not particularly limited, and can be appropriately selected from the viewpoint of its sensory and activity expression. For example, the active ingredient of the present invention in food is preferably 0.001. weight _^0/0 or more, more preferably 0.1 0001～; 10 weight _^0/0, more preferably 0.00 06-6 wt%. [0040] The method for producing the food of the present invention is not particularly limited. For example, blending, cooking, processing, etc. can be produced by a production method that follows general foods, and the obtained food contains the active ingredient according to the present invention having ACAT inhibitory action. If it is done.

 [0041] The food of the present invention contains, for example, the active ingredient of the present invention preferably from 0.1 ag to 1 adult per day.

 5 g / kg body weight, more preferably 1 g to 2 g / kg body weight, still more preferably 10 Hg to, and lg / kg body weight.

 [0042] Furthermore, the present invention provides a biological feed having an ACAT inhibitory action, comprising the active ingredient, that is, containing, adding and / or diluting the active ingredient. In addition, the present invention also provides a method for raising an organism, which comprises administering the active ingredient to the organism. Further, as another aspect of the present invention, there is provided a biological breeding agent characterized by containing the active ingredient.

 [0043] In the present specification, the organism is not limited, and examples thereof include farm animals and pet animals. Aquaculture animals include domestic animals such as horses, bushes, pigs, hidges, goats, ratadas, llamas, laboratory animals such as mice, rats, guinea pigs, and magpies, poultry such as birds, ducks, turkeys, and birds, fishes, Examples include crustaceans or shellfish. Examples of pet animals include Inu and cats. Examples of the feed include a feed for maintaining and / or improving physical condition. Examples of the animal breeding agent include an immersion agent, a feed additive, and a beverage additive.

 [0044] According to these inventions, based on the ACAT inhibitory action of the active ingredient used in the present invention in the organisms as exemplified above to which they are applied, the same as that according to the medicament of the present invention. The expression of the effect can be expected. That is, the feed of the present invention exerts a therapeutic or preventive effect on diseases requiring ACAT inhibitory action for treatment or prevention in the organism, such as arteriosclerosis and hyperlipidemia, and diseases caused by these factors. Can do.

 [0045] The active ingredient used in the present invention is usually preferably from 0.11 11 g per day to the target organism.

5 g / kg body weight, more preferably 1 g to 2 g / kg body weight, still more preferably 10 Hg to; and lg / kg body weight. For example, the active ingredient can be added and mixed in the raw material of the artificial feed for the target organism or mixed with the powdered raw material of the artificial feed and then further added to and mixed with other raw materials. It can be carried out. In addition, the effective The content of the minute in the feed yo Rere force ί column e (or ,, preferably be appropriately set according to the particular limited as those in the nag purposes <(or 0.00001 weight _^0/0 or more, more preferably <(Between 0.0001 and 10% by weight, more preferably 0.0006 to 6% by weight. The content of the active ingredient of the present invention in the biological breeding agent may be of the same level.

 [0046] The method for producing the feed of the present invention is not particularly limited, and if the formulation is similar to that of a general feed, the active ingredient according to the present invention having an ACAT inhibitory action is included in the prepared feed. It only has to be done. A biological rearing agent can be prepared in the same manner.

 [0047] In the present invention, for example, a feed containing the active ingredient used in the present invention having an ACAT inhibitory action is consumed, or a liquid containing the active ingredient used in the present invention (for example, Or maintaining the physical condition of livestock, laboratory animals, poultry, pet animals, etc., by immersing the target organism in water in which the soaking agent is dissolved in water) Power S can be. These aspects are one aspect of the method for raising organisms in the present invention.

 [0048] The active ingredient used in the present invention is not toxic even if the body is administered in an effective amount for the expression of its action. For example, in the case of oral administration, no deaths are observed even if a single dose of 3'-acetoxy-4 'tigloroxy-3', 4'-dihydroserine is administered to a mouse at lg / kg body weight. In addition, when the active ingredient is orally administered to rats, no mortality is observed even if lg / kg body weight is orally administered once.

 Example

 [0049] Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these descriptions. In the examples, “%” means “volume%” unless otherwise specified.

 [0050] Preparation Example 1 Preparation of 3'-acetoxy-4'-tigroyloxy-3 ', 4'-dihydroserine

(1) Add 100% ethanol of 2L 60% ethanol to 100g of freeze-dried button bow fu leaf stem and extract for 30 minutes with occasional stirring at room temperature. Divided into. Subsequently, extraction operation with 2 L of the same solvent was repeated on the residue. The resulting extract was collected and concentrated on a rotary evaporator. Ethanol extraction was performed several times, and the obtained extracted fraction was concentrated again. Finally, it was dissolved in 50 mL of 20% ethanol to obtain a button bow 60% ethanol extract.

 [0051] (2) Preparation Example 1 The button-bow 60% ethanol extract obtained in (1) was fractionated using reverse-phase chromatography. As the resin, Cosmo Seal 75 C18-OPN (200 mL, manufactured by Nacalai Testa) equilibrated with 20% ethanol was used. After adding button bow fu 60% ethanol oil to the moon, elution was performed in the order of 800mL 20% ethanol, 3.4L 40% ethanol, 1.6L 60% ethanol, 1L 80% ethanol. .

 [0052] (3) From the 40% ethanol eluate obtained in Preparation Example 1 1 (2), the 1.6 L to 2 L fraction and the 2 L to 2.4 L fraction were concentrated using a rotary evaporator. Each was finally dissolved in 5 mL of 40% ethanol to obtain Button Bow Fucosmosil fraction fraction 1 and Button Bow Fu Cosmosino fraction fraction 2.

 [0053] (4) Preparative Example 1 The button bow cosmosyl fraction fraction 1 and the button bow cosmoseal fraction fraction 2 obtained in (3) were fractionated using reverse phase chromatography. The force ram was TSK gel ODS-80Ts (20 mm X 25 cm, manufactured by Tosoh Corporation). The solvent was distilled water: acetonitrile = 30: 70 (volume ratio), the elution rate was 5 mL / min, and detection was performed at UV: 215 nm. The eluate was fractionated using the ultraviolet absorption of the eluate as an index.

 [0054] (5) Fractions containing a peak with a retention time of 33. 3 minutes obtained in Preparation Example 1 1 (4) were concentrated using a rotary evaporator. Finally, it was dissolved in 10 mL of dimethyl sulfoxide to obtain a button-bofu reverse phase column fraction.

 [0055] (6) Preparation Example 1 The button-bow-fu reversed-phase column fraction obtained in (1) (5) was subjected to various measurements using a nuclear magnetic resonance (N MR) spectrum apparatus (AVANCE600, Bruker Biospin). NMR spectra were measured. The mass spectrum was measured by an ion spray method using a mass spectrometer (API3000, manufactured by Applied Biosystems). As a result of NMR spectrum and mass spectral analysis, the fraction of button-boofed reversed-phase column was 3'-cetoxy-4'-tigroxy- 3 ', 4'-dihydrocereal as shown in the above formula (I). (3, 1 Acetoxy-4, — tigloyloxy— 3 ', 4, 1 dihydroseselin).

 [0056] Example 1

(1) Preparation of rat microsomes Sprague—4 Dawley rats were raised for 1 week and sacrificed. The liver was immediately removed, washed gently with cold saline, and then sucrose buffer (0.3 M sucrose, 50 mM Tris HC1, 1 mM EDTA, 50 mM NaCl, pH 7.4). ) (25 mL / l animals). The obtained liver was homogenized and centrifuged at 10,000 xg for 30 minutes to remove the precipitate twice. The obtained supernatant was centrifuged at 105,000 xg for 70 minutes to collect the precipitate, and suspended in an appropriate amount of lOOmM phosphate buffer (ρΗ7.4). The protein was suspended again in phosphate buffer so that the protein concentration became 10 mg / mL, and after adding EDTA to a final concentration of ImM, it was stored frozen at 80 ° C until use.

 [0057] (2) Measurement of ACAT inhibitory activity

 The enzyme activity of ACAT was measured with some modifications according to the method of Lee et al. (Planta Med., 2004, 70, ρ678-679).

[0058] 1. Reaction aqueous solution containing rat liver microsomes 1 · 5 L prepared in Example 1 1 (1) in a 5 mL Eppendorf tube (0 · 05 M potassium phosphate, ImM dithiothreitol, 9 mg / mL Fatty acid free serum serum albumin, 200 g / mL cholesterol, pH 7.4) 3'-acetoxy 4'-tiglooxy 3 ', 4 prepared in Preparation Example 1 to a final concentration of 14 g / mL in 92 L 'L-dihydroserine dimethyl sulfoxide solution (1 L) was added and incubated at 37 ° C for 30 minutes. In addition, as a control category, a compound was added without adding a dimethyl sulfoxide solution, and as a positive control, a compound containing FR179254 (final concentration 10 M, Merck, 344235) known to have ACAT inhibitory activity was added. Set the minute. Thereafter, 0. 05mCi / mL of Oleoyl Coenzyme A, [oleoyl- 9, 1 0- 3 H] ( Moravec Biochemicals Inc., MT1649) 2 μ L and 0 · 2 mg / mL of Ole oyl Coenzyme A (ICN 591-20521) 5 L of the mixture was added to each tube, mixed, and then incubated at 37 ° C for 5 minutes. Thereafter, the reaction was stopped by adding 0.5 mL of a solution of isopropanol: heptane = 4: 1 (volume ratio) to each tube. Add 0.2 mL of 0.1 M potassium phosphate solution (ρΗ4 · 4) and 0.3 mL of heptane to each tube, stir for 15 seconds, centrifuge for 15 seconds, and use the resulting upper layer. Radiation activity with a liquid scintillation counter LS6500 (manufactured by Beckman Co., Ltd.), using Ultimago ildo (Perkin Elma Life Science Co., Ltd., 6013329) as a scintillation basket. Sex was measured. Each reaction was performed in duplicate, and the mean value of radioactivity was calculated. The ACAT inhibitory activity (%) of the test sample was calculated by setting the inhibition rate to 100% when 10 M FR179254, which is a positive control, was added according to the following formula.

 [0059] Inhibitory activity (%) = {([Radioactivity in the control category (cpm)] [Radioactivity in the test sample addition category (cpm)]) / ([Radioactivity in the control category (cpm)] — [10 M of FR179254 calories with category Radioactivity (cpm)])} X 100

 As a result, the inhibitory activity of 3, -acetooxy-4, -tigroyloxy-3, 4, -dihydroserine was 164%, and a remarkable ACAT inhibitory activity was observed.

 [0061] According to the present invention, an ACAT inhibitor containing 3'-acetoxy-4'-tigloyloxy-3 ', 4, -dihydric mouth celine, derivatives thereof and / or pharmaceutically acceptable salts thereof, A medicament, food or feed containing it is provided. The medicament is useful as a therapeutic or prophylactic agent for arteriosclerosis, hyperlipidemia, and diseases caused by these factors. In addition, when the food is taken as a daily food, the symptoms of the above diseases can be improved or prevented. Therefore, the food containing the active ingredient of the present invention is a functional food useful for maintaining homeostasis due to its ACAT inhibitory action.

Claims

The scope of the claims  [1] One or more selected from the group consisting of 3′-acetoxy-4′-tigloyloxy-3 ′, 4′-dihydroserine, its derivatives and pharmaceutically acceptable salts thereof as an active ingredient And an acyl-CoA cholesterol acyltransferase inhibitor.  [2] A medicament comprising the inhibitor according to claim 1.  [3] A food or feed comprising the inhibitor according to claim 1.