WO2003086436A1

WO2003086436A1 - Enteric fat absorption inhibitors contaiing plant extract and foods containing the same

Info

Publication number: WO2003086436A1
Application number: PCT/JP2003/004595
Authority: WO
Inventors: Hiroshi Shimoda; Chie Yoshino; Naoki Kasajima
Original assignee: Morishita Jintan Co., Ltd.
Priority date: 2002-04-12
Filing date: 2003-04-11
Publication date: 2003-10-23
Also published as: JP3980952B2; AU2003236084A1; JP2004002254A; TW200401645A

Abstract

Enteric fat absorption inhibitors which contain a plant-origin and safe extract obtained by extracting sage and/or rosemary. These enteric fat absorption inhibitors can regulate the action of pancreatic lipase and thus inhibit the absorption of fat into the body even in a small dose. Therefore, obesity, hyperlipemia, etc. can be efficaciously treated or prevented by adding these enteric fat absorption inhibitors to various foods which can be easily taken in our everyday life.

Description

Description Enteric fat absorption inhibitor containing plant extract, and food containing it

The present invention is intended for the treatment of obesity and hyperlipidemia, which exhibits excellent knee lipase inhibitory action and fat absorption inhibitory action in the duodenum, which is the site of response, without having to suppress gastric secretion of the subject in advance. Or, it relates to an enteric fat absorption inhibitor effective for prevention.

Background art

Obesity is thought to be caused by lifestyle disorders such as lack of exercise and overeating, a genetic predisposition, and a decrease in basal metabolism.In recent years, leptin secreted by fat cells has an appetite controlling function and is closely related to obesity. It is clear that they are involved in Excessive obesity needs to be treated promptly and promptly because it can cause lifestyle-related diseases such as diabetes and cardiovascular disease (arteriosclerosis, stroke).

Obesity can be eliminated or treated by: 1. suppressing appetite, 2. suppressing the absorption of nutrients (especially lunar fat), 3. increasing thermogenesis, 4. improving the metabolism of lipids and proteins. There are extensive studies, each of which includes improving and 5. regulating central weight control mechanisms. Of these, only obesity treatments classified as 1. and 2. have been clinically applied, but because of their many side effects and their safety has not yet been established, few drugs can be used for a long time worldwide. In Japan, currently only the appetite suppressant mazindol, which is classified as 1., has been approved.

On the other hand, in addition to therapeutic agents, those that exhibit the above function 2 have been developed. For example, foods containing the ingredient having the function 2 described above in foodstuffs have attracted attention due to the recent increase in obese people and the increase in diet-consciousness. Although this does not require a prescription unlike a therapeutic drug, it is required to have as few side effects as possible, but it has the advantage that it is easily available and can be taken in everyday life.

Examples of the component having the function of the above 2 include chitin and chitosan derived from the power shell of radish and chondroitin sulfate derived from cartilage of fish, which are frequently used in health foods and the like recently. All of these were ingested due to their ability to inhibit lipase activity. Lunar fat is not enzymatically hydrolyzed, resulting in suppression of fat absorption from the intestinal tract.

Disclosure of the invention

(Technical problems to be solved by the invention)

However, since these components have a weak ability to suppress fat absorption, they had to be ingested in large amounts in a daily diet.

(How to solve it)

The present inventors have conducted research on a plant-derived safe fat absorption inhibitor capable of suppressing the action of lipase with a small amount of ingestion and exhibiting a fat absorption inhibitory effect. It was found that the extract thus obtained exhibited a superior knee lipase inhibitory effect in vitro.

Based on this finding, as a result of testing the fat absorption inhibitory effect of the extract in vivo on mice administered with olive oil, almost no fat absorption inhibitory effect was observed in the case of oral administration. It was found that when administered directly into the duodenum, or given after the secretion of gastric juice from mice was previously suppressed using a gastric secretion inhibitor, it exhibited a fat absorption inhibitory effect.

However, direct administration to the duodenum is not easy to carry out in everyday life.On the other hand, combined use with gastric secretion inhibitors, antacids, and acid neutralizers makes them less likely to digest nutrients other than fat. It is not recommended to use it regularly in your daily diet because it may affect it and may cause side effects.

On the other hand, the present inventors have proposed that the fat-absorption inhibitor containing the extract from sage or rosemary is enterically solubilized so that the extract can reach the duodenum without changing the gastric juice of the subject. It was confirmed.

Accordingly, the present invention dictates an enteric fat absorption inhibitor comprising an extract of sage and / or rosemary. The enteric fat absorption inhibitor of the present invention may be in the form of tablets, granules, powders or capsules.

Further, the present invention also provides a food containing the enteric fat absorption inhibitor.

(Effective effect than conventional technology)

According to the present invention, by applying enteric technology to a preparation containing an extract derived from a plant (sage pope rosemary), it has a lipase inhibitory activity with a small amount of ingestion. Can be effectively expressed.

The enteric fat absorption inhibitor of the present invention, when taken before or during a meal, makes the ingested fat less susceptible to enzymatic hydrolysis by knee lipase, thereby reducing the amount of fat absorbed from the intestinal tract. I can make it. Therefore, even if a high-fat diet is consumed, absorption of fat into the body can be suppressed, which is useful for the prevention and treatment of lifestyle-related diseases such as obesity, arteriosclerosis, high blood pressure, myocardial infarction, cerebral infarction, and diabetes. is there.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph showing the concentration of 50% inhibitory lipase activity inhibition (IC ₅ ) of a methanol extract from Sage tested in Example 2 of the present invention.

FIG. 2 is a graph showing the lipase inhibitory effect of each isolated component from the methanol extract of sage tested in Example 4 of the present invention.

FIG. 3 is a graph showing the inhibitory effect of carnosic'acid (2) administration on blood triglyceride elevation in mice loaded with olive oil pre-administered with a gastric secretion inhibitor tested in Example 6 of the present invention.

FIG. 4 shows the inhibitory effect on blood triglyceride elevation by administration of the methanol extract extract of sage obtained in Example 1 in olive oil-loaded mice pre-administered with the gastric secretion inhibitor tested in Example 6 of the present invention. It is a graph.

BEST MODE FOR CARRYING OUT THE INVENTION

Extractor ^ "

The enteric fat absorption inhibitor of the present invention contains an extract from Sage and Z or Masu-Zumari.

The extract from sage used in the present invention is sage, ie, a Labiatae plant

It is obtained by extracting the fresh or dried leaves of Salvia officinalis with water, a suitable organic solvent, or a solution combining them, and then concentrating. Sage is widely known as a spice in meat dishes, and has been used folk in Europe to treat diarrhea, stomach cats and intestinal catarrhs. Even now, it is considered to be highly safe for the digestive system because it is formulated in gastrointestinal drugs (EnterosanoF).

Extraction solvents used for extraction include lower alcohols such as ethanol and methanol. And lower esters such as ethyl acetate, and mixtures of these with water. Among them, it is preferable to use ethanol alone or a mixture with water (so-called hydrous ethanol) because it is consumed by humans. In particular, it is most effective to use a mixture of ethanol and water with a ratio of ethanol to water of 30% or more or 100% ethanol.

Therefore, in the examples described below, 100% methanol is used for extraction from sage and / or rosemary leaves.However, an ethanol mixture of 30% or more or 100% ethanol may be used instead. Of course it is possible. Indeed, in vitro and in vivo tests with extracts from ethanol / water mixtures containing 30% or more ethanol and active ingredients isolated therefrom have similar results to 100% methanol extracts. It has been confirmed that it exhibits an excellent fat absorption inhibitory effect and knee lipase inhibitory activity.

When sage is extracted by immersion at room temperature (15 ° C to 30 ° C), a large amount of volatile oil (digjong) and essential oils (binene, cineole, pozoleneol) contained in the sage are mixed into the extract. However, they may irritate the intestinal mucosa and adversely affect the living body. Therefore, it is preferable to extract by heating at 70 ° C or more: 100 ° C.

Regarding the amount of the solvent and the extraction time, since the plant is a leaf, it is preferable to add 5 to 20 mL of the solvent to 1 g of the plant, and to extract at the above temperature: for 3 hours.

Further, by employing supercritical carbon dioxide extraction as another extraction method, it is possible to produce an extract having a higher active ingredient content and higher safety than the solvent extraction. The extraction method may be in accordance with the method described in the literature (for example, Bauman D. et al., Acta. Mmentaria, Vol. 28, No. 1, pp. 15-28, 1999). More specifically, first, sage leaves are put into an extraction tank, and a pressure of 10 to 25 MPa (preferably 25 MPa) and a temperature of 40 to 100 ° C. (preferably 100 ° C.) are used. ~: Extraction is performed for 100 minutes (preferably 20 to 60 minutes). Under these conditions, the essential oil fraction containing harmful components is almost extracted and removed. Next, the residue is subjected to a pressure of 35 MPa or more (preferably 47.5 MPa) and a temperature of 40 to 100 ° C (preferably 100 ° C) for 20 to 100 minutes (preferably 20 to 60 minutes). Extraction is carried out to obtain an extract having a high content of diterpene fraction containing carnosic acid as an active ingredient. At this time, increase the extraction yield Therefore, an entrainer such as ethanol can be used if necessary. In the present invention, Rosemarii, ie, Lamiaceae plant Rosmarinus officinalis

Extracts obtained from fresh or dried leaves of Linne (Labiatae) may also be used. Rosemary is used not only as a spice for cooking like sage but also as a carminative and stomachic because it contains rosmarin oil, a type of essential oil. Furthermore, their use as preservatives or antioxidants is also known. Extraction can be performed in the same manner as described above for sage. That is, 5 to 20 mL of solvent is added to 1 g of fresh or dried rosemary leaf at the above temperature:! It is preferred to extract for ~ 3 hours.

The extract extracted from sage or rosemary used in the present invention may be a water or solvent extract and a supercritical diacid carbon extract obtained by the above procedure, and the extract obtained by drying the extract under reduced pressure known in the art. Includes solids obtained by removing the extraction solvent by a method such as solid or spray drying, and fractions obtained by further isolating and purifying these.

Preferred isolation and purification procedures and conditions in the present invention are described in detail in Examples 1 and 3 below, for example, a combination of silica gel column chromatography, reversed-phase ODS column chromatography, and preparative HPLC. Under suitable conditions, the reaction may be carried out using an eluent (for example, water or various organic solvents such as hexane, ethyl diester, chlorophonolem, methanol and n-butanol, or a mixture thereof). However, the present invention is not particularly limited to these. The fraction separated by the isolation and purification from the extract from Sage or Pomasum contains several types of diterpenes and triterpenes. In particular, among these, for the first time, it has been found that the carnosic acid diterpene represented by the following chemical structural formula is the most effective ingredient for suppressing fat absorption, which is the purpose of the present invention. did.

Carnosic acid, for example, in the case of sage, prepares a methanol extract from sage as described in Examples 1 and 3 below, and fractionates the ethyl acetate fraction from the sage by silica gel chromatography. Then, it is obtained through isolation and purification by ODS column chromatography and preparative HPLC.

(Fat absorption inhibitor)

The present invention relates to a fat absorption inhibitor containing an extract from sage or rosemary, which exhibits an effect of suppressing fat absorption by suppressing the action of knee lipase, particularly an extract containing carnosic'acid. Enteric coated capsules, granules or tablets (including Repetab).

As a method for producing a capsule, a conventionally known method for producing a soft capsule may be used except that the above-mentioned extract is used for the content. Examples of such a production method include a method of encapsulating the contents with a rotary filling machine using a capsule film sheet and molding a capsule preparation, or a method of producing a seamless capsule by a dropping method. .

For granules, various known granulation methods such as wet and dry methods can be applied, and the granules are molded together with appropriate binders and excipients. For tablets, a suitable binder, excipient, disintegrant and, if necessary, a lubricant are added to granules containing sage extract or sage extract itself prepared by the above-mentioned method, and the tablet is manufactured by a known tableting method. can do.

In order to impart enteric properties to the fat absorption inhibitor of the present invention, the lunar fat absorption inhibitor may be coated with an enteric 14 coating on its surface. Or turn off the fat absorption inhibitor In the case of a cell or tablet, the composition of the fat absorption inhibitor may be mixed with an enteric agent, or the composition of the coating material such as gelatin or starch may be mixed with the enteric agent to form the composition. Force capsule filling is also possible.

Enteric coating agents include celluloses such as methacrylic acid copolymers, hydroxypropynolemethylsenorellose phthalate, hydroxypropylmethylsenorellose acetate, succinate, cellulose acetate phthalate and cellulose acetate, and shellac. But are not limited to these. Such an enteric coating agent can be coated on the surface of the fat absorption inhibitor by a known method.

On the other hand, examples of the enteric agent include pectin, alginic acid, celluloses (for example, canolepox methinoresenorelose, cenorellose acetate phthalate, etc.), methacrylic acid copolymer and the like.

It is preferable to take the fat absorption inhibitor of the present invention before or during a meal. When water and Z or a solvent extract from sage or rosemary is used as the extract contained in the fat absorption inhibitor of the present invention, it is usually 10 to 500 mg, preferably 20 to 500 mg per adult. Take 200mg three times daily at meals.

In the present invention, it is also possible to use an isolated and purified fraction from the water and Z or the solvent extract as an extract. In this case, the usual dosage is 1 to 100 mg, preferably 5 to 20 mg per adult, three times a day at meals.

Of course, the intake may be appropriately increased or decreased depending on the age, the intake of fat, and the like. The intake of the fat absorption inhibitor of the present invention is 20 minutes less than that of a conventionally known natural source substance, for example, chitin 'chitosan. It can be used in amounts as small as ~ 1/2.

(Food containing enteric fat absorption inhibitor)

The enteric fat absorption inhibitor of the present invention can be added to ordinary foods and drinks to be taken on a daily basis. Particularly, it can be added to foods containing fats such as processed meats and fats and oils. preferable. Therefore, the present invention also provides a food containing such an enteric fat absorption inhibitor.

Since the food of the present invention can be ingested constantly, it can be expected to have an effect of suppressing fat absorption and is useful as a food for health use. The amount of the extract to be added in such foods and drinks may be added within a range that does not impair the original taste of the food, depending on the type of the target food, and is usually 0.1 to 10% by weight based on the target food. Just add! / ,.

Example

Next, the present invention will be further described with reference to examples, but the present invention is not limited to these examples.

Example 1 Preparation of Sedimethanol Extract and Various Solvent Fractions

Leaves (420 g) of dried Albania sage were cut into small pieces, and 10 L of methanol was added thereto, followed by extraction at 70 ° C for 3 hours. After filtration, 10 L of methanol was again added to the residue, and the mixture was extracted for another 3 hours. The filtrate obtained by two extractions was collected, concentrated under reduced pressure, and dried to obtain a methanol extract. The yield of the obtained methanol extract was 30.4%.

A portion (91 g) of the methanol extract was dispersed in 1.8 L of water, and partitioned three times with ethyl acetate (1.8 L). The obtained ethyl acetate fraction was concentrated under reduced pressure and dried to obtain an ethyl acetate fraction (48.4 g, yield: 15.8%).

Next, n-butanol (1.8 L) was added to the separated aqueous layer, and the same operation as when the ethyl acetate layer was obtained was performed to obtain the n-butanol fraction (10.3 g, yield: 3.4%). Obtained. The remaining aqueous layer was concentrated under reduced pressure and freeze-dried to obtain a water fraction (33.4 g, 11.0%). Example 2 Assay for Inhibiting Lipase of Methanol Extract of Sage and Various Solvent Fractions The lipase inhibition test was carried out using lipase derived from pig knee (Sigma) and Lipase Kit S (Dainippon Pharmaceutical). First, with respect to the methanol extract of sage and various solvent fractions (collectively, hereinafter referred to as “extracted extract”) obtained in Example 1, a dilution series of 20 times the final concentration of dimethyl sulfoxide was prepared. It was prepared. For each extract, add a coloring solution (390 wL), the dilution series (25 tL each), and a lipase solution (25 μL) prepared to 40 μg / mL using the buffer solution provided with the kit. An esterase inhibitor (lO ^ L) was prepared, and each was reacted at 30 ° C for 30 minutes. After adding a reaction stop solution (1 mL), the absorbance at 405 nm was measured, and the enzyme inhibition rate was calculated from the following equation.

P_g, (%, =] ■ (Q-P sample-Q-P -sample blank)>

(〇 ■ D ■ control ^— 0 ■ ^ ■ control blank) In the above formula, the OD _{samp le} is absorbance after enzyme treatment extract at various concentrations, the OD _{sampl e bl ank} various concentrations enzyme was not added when the absorbance of the extract _of, OD _c ontr. The absorbance at the extract was not added in and enzyme treatment, further OD _c. _{tro l} _b ! _Ank represents the absorbance when neither the extract nor the enzyme is added.

The 50% knee lipase activity inhibitory concentration ( _IC∞ ) of the extract was determined by plotting the extract concentration on the horizontal axis of the graph and plotting the inhibition rate on the vertical axis. Figure 1 shows the experimental results.

From the results in FIG. 1, it was found that the methanol extract of sage had lipase inhibitory activity (IC ₅₀ : 94 μg / mL). Further, component effective lipase inhibitory activity of the methanol extract of sage is then separated acetate Echiru fraction: was presumed to have been mainly contained in _{(IC ∞ 64 / zg / mL} ).

Example 3 Isolation and purification of sage from methanol extract

The active ingredient was isolated and purified from the ethyl acetate fraction, which was shown to have concentrated lipase inhibitory activity in Example 2.

The above ethyl acetate fraction (20.1 g) was subjected to silica gel column chromatography (600 g), and hexane: ethyl acetate (10: 1) → (3: 1) → (1: 1) → chloroform: Elution was performed in the order of methanol (10: 1) → methanol to obtain a fraction 1 (4.22 g), a fraction 2 (4.25 g), a fraction 3 (3.3 g), and a fraction 4 (7.86 g). For these fractions, the knee lipase inhibition rate at 100 g / mL was measured using the method of Example 2 and found to be 43.4, 96.0, 92.6 and 67.7%, respectively.

Fractions 2 and 3, which had a high knee lipase inhibition rate, were separated by reversed-phase ODS column chromatography [220 g, water: methanol (4: 6) → (3: 7) → (2: 8) → (1: 9) ) → Cross-form: methanol: water (6: 4: 1)] and preparative HP LC [ODS, solvent 75% methanol].

The isolated component was analyzed by ¹³ C NMR analysis, mass spectroscopy, infrared spectroscopy, and ultraviolet-visible spectroscopy. Carnosol (in the structural formula shown below, (1) Represented, Yield from plant: 0.24%), Carnosic acid (Id. (2), 0.29%), 7-Methoxylozmanol (Id., (3), 0.081%), Loureanoitsk-acid (same, (4), 0.011%), Rosmanol (same, (5), 0.006%), Isolozmanol (same, (6), 0.027%) and oleanolic acid (same as above) , (7), 0.47%).

(5) (6) (7)

Example 4 Lipase inhibitory activity of each isolated component from sage methanol extract Extract The lipase inhibitory activity of each isolated component obtained in Example 3 was examined using the method of Example 2. The result is shown in figure 2.

As shown in FIG. 2, Jitenorepen of carnosol (1) force the strongest lipase inhibitory activity _{(IC 50: 4 / ig /} mL) indicates, carnosic 'acid (2), 7- Metokishiro Zumanoru (3), Roireanoitsuku - Acid (4) also showed inhibitory activity (IC ₅₀ : 14, 33 and 50 / zg / mL), respectively. In addition, it was found that it has a lipase inhibitory activity even though it is weak against the triterpene oranolic acid (7).

For rosmanol (5) and isolozmanol (6), lipase-inhibitory activity could not be measured because of the marked degradation due to oxidation.

Example 5 Methanol Extract of Sage and Fat Absorption Inhibition of Each Isolated Component The methanol extract of sage obtained in Example 1 and the components obtained in Example 3 (here, carnosol (1), carnosic ■ Acid (2), 7-methoxy rosmanol (3), and oleanolic acid (7)) inhibit the absorption of fat by triglyceride in the blood by oral administration or duodenal administration to mice loaded with olive oil. It was evaluated by measuring the change in concentration. (Oral administration) The methanol extract or each isolated component was suspended in water together with 5% arabia gum to prepare an aqueous suspension. The ddY male mice fasted for 20 to 24 hours were orally administered with the water suspension at a rate of 5 mL / kg body weight. Thirty minutes later, oral oil was orally administered at a rate of 5 mL / kg body weight, and two hours later, blood was collected from the orbital vein to measure the blood triglyceride concentration.

(Duplicate administration) Using a different mouse from the above-mentioned oral administration test system, the effect of suppressing fat absorption when administered directly into the duodenum was examined. The mice were laparotomized under ether anesthesia, and the aqueous suspension was directly administered into the duodenum at a ratio of 5 mL / kg body weight. After suturing the incision, 30 minutes after intraduodenal administration, olive oil was orally administered at a rate of 5 mL / kg body weight. Two hours later, the blood triglyceride concentration was measured.

Table 1 summarizes the results of the oral and intraduodenal experimental systems for the methanol extract of sage (dose: 250 or 500 mg / kg), and the four components obtained in Example 3 (here, The results for carnosol (1), carnosic acid (2), 7-methoxy rosmanol (3) and oleanolic acid (7) are shown in Tables 2 and 3, respectively.

¾ L: Effect of administration of methanol extract from sage on blood triglyceride elevation in olive oil-loaded mice

Blood triglyceride

(mg / dL)

Oral administration Intraduodenal administration Untreated group 150 ± 17 * * 117 ± 23

Olive oil administration group 409 ± 75 204 ± 25

Sage 383 ± 25 134 ± 15

Methanore extract (250mg / kg)

+ Olive oil administration group

Sejimethanol extract (500mg / kg) 395 ± 35 118 ± 15

+ Olive oil administration group 2: Effect of administration of each isolated component from sage methanole extract on blood triglyceride elevation in olive oil-loaded mice

Blood triglyceride

(mg / dL)

Oral administration Duodenal administration Untreated group 181 ± 18 128 ± 17

Olive oil administration group 587 ± 43 193 ± 25

Carnosol (100 mg / kg) 473 ± 58 169 ± 23

+ Olive oil administration group

Carnosol (200mg kg) 514 ± 36 175dz23

+ Olive oil administration group

Canolenosic '' Acid (50mg / kg) 292 ± 41 143 ± 14

+ Olive oil administration group

Canolenosic 'acid (lOOmg / kg) 265 ± 30 * * 97 ± 21 **

+ Olive oil administration group

Power Glenodic Acid (200mg / kg) 234 ± 25 ** 20 ± 3 **

+ Olive oil administration group

Table 3: Effect of administration of each isolated component from sage methanole extract on blood triglyceride elevation in mice loaded with ori

Blood triglyceride

(mg / dL)

Oral administration Intraduodenal administration Untreated group 184 ± 28 131 ± 11

Olive oil administration group ±± 177 177 ± 17

7-Methoxyrosmanol (200mg / kg) 438 ± 51 245 ± 38

+ Olive oil administration group

Oleanolic acid (200mg kg) 528 ± 76 178 ± 27

+ Olive oil administration group

Each value was shown as the average value of seven animals and the standard error. ** indicates a significant difference from the olive oil administration group: p <0.01.

From the results in Table 1, it was found that when the methanol extract of sage was orally administered, no inhibitory effect on blood triglyceride elevation was observed at all, but that an intraduodenal administration exhibited an inhibitory effect on blood triglyceride increase. In other words, it was revealed that the methanol extract of Sage abolished knee lipase activity in the stomach.

The results in Table 2 show that carnosol showed the strongest lipase-inhibiting activity in Example 4. (1) Potassium tridaricella in both oral and intraduodenal administration. No inhibitory effect on id elevation (that is, fat absorption inhibitory effect) and canolenogic ■ acid (2) 1 Shows a dose-dependent inhibitory effect on triglyceride increase by oral administration, and the effect is further enhanced by intraduodenal administration It is shown to be done. From Table 3, it was found that neither the oral administration nor the duodenal administration of 7-methoxyrosmanol (3) dioleanoleic acid (7) exhibited the triglyceride elevation inhibitory effect.

From the above results, the lipase inhibitory effect of the methanol extract of sage confirmed in Example 2 is strongly related to the fat absorption inhibitory effect of carnosic'acid (2), which is inactive in the stomach. It turned out to be.

Example 6 Inhibition of blood triglyceride elevation by administration of carnogic acid in olive oil-loaded mice pre-administered with gastric secretion inhibitor

According to the results of Example 5, the fat of carnosic acid (which is represented by (2) in the formula of Example 3; hereinafter, referred to only as (2)), which is an isolated component effective for the lipase inhibitory action, was obtained. The effect of gastric juice on the ability to suppress fat absorption was examined in more detail regarding the fact that the absorption suppressing ability was inactivated in the stomach.

The procedure is as follows.

(ϋ) After 30 minutes, an aqueous suspension of 5% gum arabic and carnosic 'acid ((2), 100 mg / kg) suspended in water is orally administered at a rate of 5 mL / kg body weight. And

(iii) After further 30 minutes, the olive oil was orally administered at a rate of 5 mL per kg of body weight.

(iv) Two hours after (iii), blood was collected from the orbital vein and the blood triglyceride concentration was measured.

The results are shown in Figure 3.

In Figure 3,

Column 1 shows the untreated group (system in which only step (iv) was performed).

Column 2 shows the group receiving olive oil (that is, the system in which only steps (iii) to (iv) were performed), and column 3 shows the group receiving carnosic'acid ((2), 100 mg kg) + olive oil (group Other than the process すなわち, that is, the systems (ii) to (iv)],

Column 4 shows the groups administered with cimetidine and olive oil [systems that performed (1) and (iii) to (iv) except for step (ii)], and

Column 5: Cimetidine + Carnosic 'Acid ((2), 100 mg / kg) + Olive oil administration group [to (iv) all steps]

Are respectively shown. Each column is shown by the average value and standard error of 7 mice.

In FIG. 3, * represents a significant difference from the olive oil administration group (column 2): p く 0.05, and ** represents a significant difference: p く 0.01.

The blood triglyceride concentration of mice (column 5) to which carnosic acid (2) was administered after gastric secretion was previously suppressed using cimetidine was similar to that of carnosic 'acid (2) without addition of gastric juice secretion inhibitor. Mice administered alone

(Column 3). From this, it was found that carnosic acid ( ₂₎ , which had a fat absorption inhibitory effect, lost its activity by gastric juice.

For confirmation, the same evaluation was performed for the methanol extract of sage obtained in Example 1. Fig. 4 shows the results.

In Figure 4,

Column 2 shows the olive oil administration group (that is, the system where only steps (iii;) to (ίν) were performed), and column 3 shows the sage methanol extract (500 mg / kg) + olive oil administration group (except for step I). That is, (ii) to (iv))

Toram 4 is a group administered with cimetidine and olive oil [systems (i) and (iii) to iv) other than step (ϋ)], and

Column 5: Cimetidine + sage methanol extract (500 mg / kg) + olive oil administration group [① to (iv) all steps]

In FIG. 4, ** indicates a significant difference from the olive oil administration group (column 2): p <0.01.

Comparison of columns 3 and 5 in Fig. 4 indicates that cimetidine inhibits gastric juice secretion in the sage methanole extract containing carnosic acid (2). This indicates that the effect of suppressing fat absorption is enhanced.

Example 7 Preparation of Sage Supercritical Carbon Dioxide Extract

100 g of dried sage leaves were charged into the extraction tank, and extraction was performed for 60 minutes by supplying critical carbon dioxide at a pressure of 25 MPa and 100 ° C. Subsequently, the residue was extracted under conditions of a pressure of 47.5 MPa and 100 ° C. for 60 minutes to obtain a sage supercritical carbon dioxide extract (7 g). Example 8 Fat Absorption Inhibitory Effect of Sage Supercritical Carbon Dioxide Extract

The fat absorption inhibitory effect of the sage supercritical carbon dioxide extract obtained in Example 7 was examined by the same method as in Example 5. Table 4 shows the results.

¾4: Administration of sage supercritical carbon dioxide extract suppresses blood triglyceride elevation in olive oil-loaded mice

Blood triglyceride

(mg / dL)

Oral administration Duodenal injection

Giving

Untreated group 144.5 ± 88 131.2 ± 10.9

Olive oil administration group 619.8 ± 74.1 176.9 ± 17.2

Sage supercritical carbon dioxide extract 403.8 ± 73.4 ** 140.9 ± 20.1

(50mg kg

+ Olive oil administration group

Sage supercritical carbon dioxide extract 230.2 ± 38.0 * * 125.5 ± 10.4

(100mg / kg) + olive oil administration group

Each value was shown as the average value of seven animals and the standard error. ** indicates a significant difference from the olive oil administration group: p-0.01.

From this result, it was clarified that the supercritical carbon dioxide extract of sage had a very strong fat absorption inhibitory action.

Formulation Example 1 (Preparation of enteric capsule formulation containing methanol extract of sage)

Using the compositions and weight ratios shown in Table 5 below, enteric capsule preparations containing sage methanol extract (powder) were produced according to the following procedure. The sage methanol extract (powder) is obtained by pulverizing the dry solid obtained in Example 1 using a mill or a blender.

First, a methanol extract of sage (powder) was suspended in hardened fat (hardened fat) at 40 ° C to prepare capsule contents. Separately, gelatin, glycerin and vectin were mixed to prepare a capsule outer coating solution. Next, in the capsule manufacturing machine, cooling is performed by flowing the capsule contents from the innermost nozzle of the concentric triple nozzle, hardened oil and fat from the intermediate nozzle as the substance forming the inner capsule, and the outer capsule liquid from the outermost nozzle. The triple-layer seamless force capsules were continuously manufactured by simultaneously discharging them into the liquid oil thus obtained. The particle size of the obtained seamless force capsule could be freely adjusted from 1 mm to 8 mm.

The enteric solubility of the obtained seamless capsule was tested by the collapse test described in the Japanese Pharmacopoeia. As a result, the seamless capsule did not collapse within 2 hours in the first liquid (pH 1.2) and then collapsed within 60 minutes in the second liquid (pH 6.8). It was confirmed to be enteric.

Formulation Example 2 (Preparation of enteric-coated forceps containing methanolic extract of rosemary) A methanol extract of mouth somary was prepared according to the procedure of Example 1 except that rosemary was used instead of sage. The dried solid of the obtained methanol extract was pulverized using a mill or a blender, and the same amount was used in place of the sage methanol extract powder in Table 5 above, and the same procedure as in Preparation Example 1 was used. An enteric capsule formulation containing a methanol extract of rosemary (powder) was produced. The obtained capsule was tested in the same manner as in Formulation Example 1 in accordance with the collapse test of the Japanese Pharmacopoeia, and it was confirmed that it was enteric.

Claims

The scope of the claims

1. An enteric fat absorption inhibitor containing an extract from sage and / or rosemary.

2. The enteric fat absorption inhibitor according to claim 1, which is in the form of a fixed agent, a granule or a capsule.

3. The enteric fat absorption inhibitor according to claim 1, wherein the extract contains carnosic acid represented by the following formula.

4. A food containing the enteric fat absorption inhibitor according to any one of claims 1 to 3.