JP2001231500A - Lipase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a lipase inhibitor having high safety and more superior lipase-inhibiting activities. SOLUTION: This lipase inhibitor contains a plant body of Myrcia sphaerocarpa, Myrcia multiflora or an extract thereof. The pharmaceutical composition for therapy of a disease for which a suppressor of lipid level in the blood, and the inhibition of the lipase activities are effective for the cure, (e.g. obesity, diabetic, hypertension, hyperlipemia, arteriosclerosis and acne) is also provided.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to Myrcia sphaerocarpa.
lipase inhibitor containing carpa , Myrcia multiflora (generic name: Pedra ume carr) or an extract thereof as an active ingredient, and more specifically, a food and medicine useful for lipid-lowering or anti-obesity containing the lipase inhibitor It is about.

2. Description of the Related Art In recent years, in Japan, the intake of nutrient-rich meals, for example, animal protein, animal fat, instant food, fast food, and the like has been remarkably increased with the westernization of dietary habits. As a result, calorie intake has increased, and obesity has significantly increased due to excessive nutrition due to overeating and lack of exercise. Obesity is a condition in which fat is excessively stored and accumulated in the body, including subcutaneous adipose tissue, and a BMI value of 25 or more is generally regarded as obesity.

[0003] Obesity is considered to be one of the risk factors for diabetes, hypertension, arteriosclerosis and hyperlipidemia, since the underlying pathology of obesity is hyperinsulinemia and the tendency to accumulate fat associated therewith. In addition, excessive fat accumulation in the body limits physical activity in the obese person and places extra burden on the internal organs, which may cause complications such as hyperuricemia, cholelithiasis, and gout. Many are serious problems.

[0004] Treatment of obesity generally includes dietary therapy with calorie restriction or exercise therapy with energy expenditure and combinations thereof. However, these methods need to be continued for a long period of time, which requires patience and is mentally burdensome, and results in a temporary loss of weight, which increases the possibility of weight rebound. is there. Also, in surgery and pharmacotherapy,
Surgery is not common for patients with severe obesity. As for pharmacotherapy, several drugs have been developed, for example, an appetite suppressant such as an adrenergic agonist such as mazindol or phentermine or a serotonin agonist such as dezfenfluramine or fluoxetine. In some cases, overeating due to stress causes obesity. In this case, an autonomic nervous regulator such as tofisepam may be used. However, these drugs are used only for highly obese patients who have difficulty in dieting, and there are problems in that the indications are limited in some cases, and in terms of the effects and side effects of the drugs.

On the other hand, lipase is a digestive enzyme for lipids,
In humans, it is secreted from the pancreas and Candida cyli
It is produced by microorganisms resident on the skin surface represented by ndraccae. By this lipase, ester bonds of triacylglycerol are sequentially hydrolyzed, and glycerol and fatty acids are generated via diacylglycerol and monoacylglycerol. These are absorbed from the small intestine, but the absorption of glycerol and fatty acids due to excessive fat intake leads to obesity. As described above, lipase is closely related to obesity, hyperlipidemia, arteriosclerosis, etc., and is also related to acne, etc., and thus, lipase inhibitors are used to treat these diseases and complications. It is considered effective for prevention.

Until now, as lipase inhibitors, extracts of 14 kinds of crude drugs such as peonies, oren, psyllium, buttonpig, genoshoko, tea and the like (Japanese Patent Laid-Open No. 64-901)
No. 31) or a polar or non-polar solvent of pepper, pumpkin, shimeji, maitake, hijiki, green tea, black tea, water extract of oolong tea (Japanese Unexamined Patent Publication No. 3-219872), dokkatsu, ryokyo, areca, biloba, sampens and beetroot Extract (JP-A-5-255)
No. 100), a lipase inhibitor containing hinokitiol as an active ingredient (Japanese Unexamined Patent Publication No. 8-268882), grape seeds, oyster leaves, puer tea, hypericum, apples, cod, urajigashi, banaba leaves, akamegashiwa, sanshuyu, mako, eucommia Leaf extract (JP-A-9-22739)
No. 8), a tamarind seed coat extract containing procyanidin as an active ingredient (JP-A-9-291039) and the like have been reported. However, their efficacy was still unsatisfactory. Therefore, it can be said that a drug useful for hyperlipidemia and obesity and having few side effects has been desired.

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to provide a lipase inhibitor which is highly safe and has better lipase inhibitory activity.

The present inventors have found that Milkia cephaerocarpa, a plant distributed in South America such as Brazil, has lipase inhibitory activity, and have completed the present invention. Milkia cephaerocarpa was not known to inhibit lipase activity at all.

According to the present invention, there is provided a lipase inhibitor comprising a plant of Milkia cephaelocarpa or an extract thereof.

According to the present invention, a blood lipid concentration inhibitor comprising a plant of Milkya cephaelocarpa or an extract thereof, and a disease in which inhibition of lipase activity is therapeutically effective (eg, obesity) , Diabetes, hypertension, hyperlipidemia, arteriosclerosis, comedones).

According to the present invention, there is provided a health food comprising a plant of Milkya cephaelocarpa or an extract thereof.
In particular, a food is provided for the purpose of suppressing blood lipid concentration or anti-obesity.

The plant of Milkia cephaerocarpa and its extract have excellent lipase inhibitory activity. Therefore, by using a plant of Milkya cephaerocarpa or an extract thereof as a lipase inhibitor, an anti-obesity agent or a health food or the like, obesity, diabetes, hypertension, hyperlipidemia, arteriosclerosis or the like or Complications can be treated.

[0013]

DETAILED DESCRIPTION OF THE INVENTION The lipase inhibitor according to the present invention comprises
It is characterized by comprising a plant of Milkia cephaerocarpa or an extract thereof as an active ingredient.

Milkia cephaerocarpa is a plant distributed in the South American region such as Brazil, and its plant body, preferably its leaves, can be used. The plant of Milkia cephaerocarpa may also be processed according to a conventional method such as pulverization, repair, roasting, drying, and fermentation. In the present specification, the term “plant” is used in a meaning including a processed one such as pulverization, repair, roasting, drying, and fermentation.

"Extract" of Milkia Cephaerocarpa
Is an extract obtained by extracting Milkia Cephaerocarpa with a polar solvent or edible oil, its diluted solution, concentrated solution,
It means an extract and a dried product obtained by drying it. Here, the "polar solvent" includes water; alcohols such as methanol, ethanol, 1-propanol, 2-propanol and 1-butanol; ethers such as 1,4-dioxane; ketones such as acetone; Nitriles; mixtures of these;
Water; lower alcohols such as methanol and ethanol; and mixtures thereof, particularly preferably water, ethanol, and mixtures thereof.

The extract of Milkia cephaerocarpa is:
A polar solvent is added to a plant of Milkia cephaelocarpa or a dried product thereof, preferably a leaf or a dried product of leaves, obtained by pulverizing, crushing, or cutting the same, and then added at 0 ° C to 100 ° C, preferably at room temperature to 70 ° C. ~ 24 hours, preferably 5-
It can be obtained by leaving it for 10 hours, removing the insolubles by filtering, centrifuging or the like as necessary, removing the solvent, and concentrating. Further, if necessary, it can be purified by washing with a non-polar solvent such as ethyl acetate, diethyl ether, and hexane, and extracting with water.

As used herein, "lipase" refers to an enzyme that hydrolyzes glycerol esters and releases fatty acids, and includes, for example, pancreatic lipase and bacterial lipase produced by microorganisms.

The plant of Milkia cephaerocarpa or an extract thereof suppresses an increase in plasma triglyceride level in rats in a dose-dependent manner (Example 3). Therefore, according to the present invention, a blood lipid concentration inhibitor is provided.

According to the present invention, there is also provided a pharmaceutical composition for treating a disease in which inhibition of lipase activity is therapeutically effective.

Diseases in which inhibition of lipase activity is therapeutically effective include obesity, diabetes, hypertension, hyperlipidemia, and arteriosclerosis (Journal of American Association., 281,
235 (1999)) and comedones.

The administration form of the pharmaceutical composition according to the present invention is not particularly limited, and examples thereof include general administration routes such as oral administration, rectal administration, transdermal administration, and administration by injection.

The plant of Milkya cephaerocarpa or an extract thereof can be formulated into a suitable solid preparation or liquid preparation using a pharmaceutically acceptable carrier depending on the administration route.

The solid preparation for oral administration includes capsules, tablets, pills, troches, powders, granules and the like. Solid preparations can generally be prepared by mixing the plant of Milkya cephaerocarpa or an extract thereof with at least one additive (eg, crystalline cellulose, lactose and starch). This formulation may be prepared with a lubricant such as magnesium stearate in addition to the additives. In the case of capsules, tablets and pills, a buffer may further be used. Tablets and pills can also be provided with an enteric coating.

Liquid preparations for oral administration include inert diluents usually used for preparing liquid preparations, such as water,
And pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. Liquid preparations are prepared by adding auxiliary substances such as wetting agents, emulsifying and suspending agents, and seasonings and flavoring agents in addition to the plant of Milkya cephaerocarpa or its extract and additives. Can be prepared.

Examples of the form of the external preparation include ointments, creams, lotions, cataplasms and the like. An external preparation can be prepared generally by mixing a plant of Milkya cephaerocarpa or an extract thereof with at least one additive (eg, gelled hydrocarbon, higher fatty acid ester, white petrolatum, etc.).

The form of the injection may be a pharmaceutically acceptable sterile or non-aqueous system, suspension or emulsion. Examples of suitable non-aqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Such compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents.
These compositions may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporation of a sterilizing agent immediately before use, in the form of a sterile solid or a sterile solid composition that can be dissolved in some other sterile injectable medium. Can be.

[0027] Formulations for rectal administration include Milkia®
It can be prepared by adding an excipient (for example, cacao butter or suppository wax) to a plant of Cephaerocarpa or an extract thereof.

The dosage of the plant of Milkya cephaerocarpa or its extract varies depending on the method of administration and the purpose of use, but it is usually 0.5 to 10 g at a time, 0.15 to 30 g as a daily dose, Preferably, it is 1 to 6 g at a time, and 3 to 18 g, more preferably 3 to 10 g as a daily dose. When these solutions are used, 0.1 to 2 wt%
The solution may be 50 to 200 ml in three doses.
As an injection, 10 to 500 ml, preferably 100 to 300 ml may be administered at a time as a 0.5 to 2 wt% solution.

In the present specification, “treatment” is used to mean “prevention”. The plant of Milkia cephaerocarpa or an extract thereof can be the food itself. The plant of Milkia cephaerocarpa or an extract thereof may also be added to foods, especially foods containing lipids (eg meat products and edible oils). Foods comprising a plant of Milkya cephaelocarpa or an extract thereof, and foods to which a plant of Milkia cephaelocarpa or an extract thereof are added, for example, health foods for the purpose of preventing obesity, prevention of diabetes, It can be a health food for the purpose, a health food for the purpose of preventing hyperlipidemia, a health food for the purpose of preventing hypertension, and a health food for the purpose of preventing arteriosclerosis. When used as foods, solid foods, creamy or jam-like semi-liquid foods, gel foods, beverages, drinks, tea leaves and any food form, for example, powder, capsules,
Examples include granules, tablets, drinks, tea bags and the like. Such foods can be processed according to conventional methods.

[0030]

EXAMPLES The present invention will be described below by way of examples, which do not limit the present invention. The following experiment was performed to evaluate the pancreatic lipase inhibitory effect of the Milkia cephaerocarpa extract.

Example 1: Preparation of lipase inhibitor Milkia cephaerocarpa leaves (HIROBRA'S IMPORT, EXP
44 g (obtained from ORT & REP. LTD.) Was crushed in a blender together with 1 liter of 30% (v / v) ethanol and left at room temperature for about 160 hours to extract a lipase inhibitor. After filtration, 1 liter of 30% (v / v) ethanol was added to the residue, and the mixture was allowed to stand at 68 ° C. for 8 hours to extract a lipase inhibitor. After the filtrates were combined and the solvent was distilled off, lyophilization was performed to obtain a dry extract. The yield was about 23%.

Example 2 Lipase Inhibition Test As a lipase substrate solution, 2% soybean oil emulsified with a 0.5% gum arabic aqueous solution was used. Substrate solution 0.1 ml, Sigma pancreatic lipase 42 unit, test sample 0.05
ml, 1 M Tris-HCl buffer (pH 7.7) 0.05 m
1 and water were added to make a total volume of 0.5 ml. Pancreatic lipase was added and allowed to react at 37 ° C. for 2 minutes, and a substrate solution was added and reacted for another 10 minutes. The reaction was stopped by adding a copper reagent (0.45 M triethanolamine, 0.05 M acetic acid, 3.4% copper sulfate pentahydrate, 20% sodium chloride),
Heptane: chloroform mixture (3: 2, v / v) 1.5
ml was added for extraction. After centrifugation, 1 ml of the organic solvent layer was taken, mixed with 1 ml of a 1% butanol solution of 0.1% diethyldithiocarbamate, and the absorbance at 440 nm was measured. Instead of pancreatic lipase, a known amount of oleic acid (0-
(1.0 μmol) was treated in the same manner to prepare a calibration curve. The same reaction was carried out using water instead of the test sample as a control, the ratio (%) to the control in the presence of the test sample was calculated, and the inhibitory effect was evaluated. As a result, 30 of Milkia Cephaerocarpa leaves
% Ethanol extract showed an inhibitory effect. The extract concentration (IC ₅₀ ) that inhibited the activity of the control by 50% was 2.9 μg / ml. The results were as shown in FIG.

Example 3 Soybean Oil Tolerance Test Male ICR mice (6 weeks old) were orally administered with water or a test sample, and immediately with 5 ml / kg of soybean oil. Blood was collected from the fundus vein before administration and 1.5, 3, and 4.5 hours after administration, and plasma triglyceride was quantified using a “Triglyceride G-Test Wako” (manufactured by Wako Pure Chemical Industries) kit. As a result, Milkia Cefaerocarpa leaves 3
In the group to which the 0% ethanol extract was administered, the increase in plasma triglyceride level was suppressed in a dose-dependent manner. The results were as shown in FIG.

Example 4: Acute toxicity test Male ddY mice (6 weeks old) were fasted overnight and 0.5%
The test substance suspended in carboxymethylcellulose was orally administered (5 animals). Immediately after and 2 hours after administration, the general condition was observed, and once a day from the next day, the general condition was observed for 7 days. Surviving animals were necropsied on the day of observation. as a result,
At the dose of 8.9 g / kg, no abnormalities were observed in all cases.

[Brief description of the drawings]

FIG. 1 is a dose inhibition curve of lipase inhibition by a 30% ethanol extract of Milkia cephaerocarpa leaves.

Fig. 2. 30 leaves of soybean oil and Milkia Cephaerocarpa leaves
FIG. 3 is a graph showing the time course of plasma triglyceride level of mice to which a% ethanol extract was orally administered.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 43/00 111 A61P 43/00 111 C12N 9/99 C12N 9/99 // (C12N 9/99 (C12N 9/99 C12R 1:91) C12R 1:91) (72) Inventor: Toshinobu Morita 1624 Shimodakodate, Koda-cho, Takada-gun, Hiroshima Prefecture F-term (in reference) 4B018 LB08 LE01 LE02 LE03 MD14 MD61 ME03 ME04 MF01 MF06 4C088 AB57 AC01 BA10 MA52 ZA42 ZA45 ZA70 ZC20 ZC33 ZC35

Claims (6)

[Claims] (1) Milkia cephaerocarpa ( Myrcia sph)
aerocarpa , Myrcia multiflora ) or a lipase inhibitor comprising the extract thereof. 2. Milkia cephaerocarpa ( Myrcia sph)
aerocarpa , Myrcia multiflora ) or an extract thereof. 3. Milkia cephaerocarpa ( Myrcia sph)
A pharmaceutical composition comprising a plant of aerocarpa , Myrcia multiflora ) or an extract thereof, and a pharmaceutically acceptable carrier, which is used for treating a disease in which inhibition of lipase activity is therapeutically effective. 4. The composition according to claim 3, wherein the disease in which inhibition of lipase activity is therapeutically effective is selected from obesity, diabetes, hypertension, hyperlipidemia, arteriosclerosis, and comedone. . (5) Milkia cephaerocarpa ( Myrcia sph)
aerocarpa , Myrcia multiflora ) or a health food comprising an extract thereof, which is intended to prevent obesity, diabetes, hyperlipidemia, hypertension, arteriosclerosis, or slimming. 6. A milky cephaerocarpa ( Myrcia sph)
A health food comprising a plant of aerocarpa , Myrcia multiflora ) or an extract thereof, and lipid or a food containing lipid.