JP2018008924A - Tie2 active composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide means for activating Tie2 conveniently to improve a blood flow and improves a lymph flow, and also improves various symptoms of the body.SOLUTION: According to the present invention, there is provided a Tie2 activating composition comprising a persimmon leaf extract.SELECTED DRAWING: None

Description

  The present invention relates to a composition for Tie2 activity.

  Tie2 (endothelium-specific receptor tyrosine kinase 2) is a receptor tyrosine kinase expressed in vascular endothelial cells and has recently been shown to be deeply involved in aging. Furthermore, it has been found that it acts on skin epidermis cells, chondrocytes, etc. in relation to angiogenesis, and is also involved in lipid, sugar, and energy metabolism. Attention has been paid.

  Tie2 is known to strengthen the connection between endothelial cells / mural cells and endothelial cells and contribute to the stabilization of the structure of blood vessels and lymphatic vessels. Since Tie2 decreases (inactivates) due to aging and disease, nutrients leak from blood vessels, especially in middle-aged and elderly people, and waste and water are accumulated in the body without being collected from leaking from lymphatic vessels. Adverse effects such as swelling and stiffness of the body and coldness of limbs are likely to occur.

  For this reason, methods and materials for simply activating Tie2 are required, and materials that suggest various Tie2 activations have been studied (for example, Patent Documents 1 to 6).

JP 2011-102273 A JP 2011-102274 A JP 2011-102275 A JP 2013-241356 A JP 2014-097977 A Japanese Patent Laying-Open No. 2015-168656

  An object of the present invention is to provide a means for easily activating Tie2. Another object of the present invention is to improve blood flow and lymph flow, thereby improving various symptoms of the body.

  The present inventors have found that the koji leaf extract has the ability to activate Tie2, and have made further improvements to complete the present invention.

The invention encompasses, for example, the subject matter described in the following sections.
Item 1.
The composition for Tie2 activity containing a persimmon leaf extract.
Item 2.
Item 2. The composition according to Item 1, comprising a koji leaf extract and a lemon myrtle extract.
Item 3.
Item 3. The composition according to Item 1 or 2, which is an oral composition.
Item 4.
Item 4. The composition according to any one of Items 1 to 3, which is a pharmaceutical composition or a food composition.
Item 5.
A method for producing a composition for Tie2 activity, comprising a step of subjecting persimmon leaves or persimmon leaves and lemon myrtle to extraction with water, ethanol, or a water-ethanol mixture.
Item 6.
A dried tea leaf or tea bag comprising dried bamboo leaves and dried lemon myrtle.
Item 7.
Dry tea leaves or tea bags for Tie2 activity, including persimmon leaves, persimmon leaves and lemon myrtle.
Item 8.
Item 5. The composition according to any one of Items 1 to 4, which is for swelling improvement, blood flow improvement, lymph flow improvement, body stiffness improvement, cold improvement, or skin wrinkle, sagging or roughness improvement.
Item 9.
Including leaf extract, or leaf extract and lemon myrtle extract for swelling improvement, blood flow improvement, lymph flow improvement, body stiffness improvement, cold improvement, or wrinkle, sagging or roughening improvement ,Composition.
Item 10.
Dried tea leaves, including dried koji leaves or dried koji leaves and dried lemon myrtle, for swelling improvement, blood flow improvement, lymph flow improvement, body stiffness improvement, cold improvement, or wrinkle, sagging or roughening improvement Or tea bags.

  According to the koji leaf extract-containing composition (preferably the koji leaf extract and the lemon myrtle extract-containing composition) according to the present invention, Tie2 can be easily activated. In addition, blood flow improvement and lymph flow can be improved easily, and thus various symptoms of the body can be improved.

The Tie2 activation examination result when using a combination of a herb extract and a herb extract that gives lemon flavor is shown. * Indicates p <0.05 (vs. leaf extract). The cell viability examination result when using a combination of a herb extract and a herb extract that gives lemon flavor is shown. * Indicates p <0.05 (vs. leaf extract).

  The composition according to the present invention includes a koji leaf extract. More preferred is a composition comprising a koji leaf extract and a lemon myrtle extract.

  The koji leaf extract is preferably an extract obtained from koji leaf by extraction with water, ethanol or a water-ethanol mixture.

  柿 is a deciduous tree of the genus Oysteraceae. The leaves that are preferably used for extraction are preferably leaves (especially young leaves) collected from trees before falling. The variety is not particularly limited as long as it is a persimmon, but specifically, the sweet varieties include Tomiyu, Jiro, Amami Hundred Sun, Gosho, Hana Gosho, Yogo Gosho, Tenjin Gosho, Fujiwara Gosho, Tokuda Gosho , Mitagaya Imperial Palace, Zenjimaru, Fujihachi, Mizushima, New Year and so on. As a variety of fruit and vegetables, Yokono, Hirakawa Mu, Fuji, Saijo, Dogami Beeya, Aizu Michichi, Kinmon, Gionbo, Yotsumizo, Dai Yotsumizo, Atago, Hagakushi, Kawabata, Takura, Sakushu Mischief, etc. Is mentioned.

The bamboo leaves may be used for extraction as they are after being collected from the tree, but may be subjected to extraction after pretreatment according to a known method. In particular, the koji leaf used in the present invention is preferably a dried koji leaf, more preferably a koji leaf that has been dried without undergoing a heating step (for example, a koji leaf that has been dried by sun drying or natural drying). For example, after sampling, it is washed with water and is of an appropriate size (usually 3 mm
After being cut into widths), it can be dried by steaming with steam. You may cut | disconnect immediately after extract | collecting and dry it according to a conventional method. Or like normal green tea, what passed through each process, such as steamed leaves, twisting, and roasting, may be used.

  Lemon Myrtle (Backhouse citriodora) is used as a tea leaf for herbal tea, as well as being used for cooking and medicinal herbs by Aboriginal Australian natives. As a site | part of the lemon myrtle used for extraction, a leaf (a leaf axis and a petiole are included) and a branch are preferable, for example, and a leaf is more preferable. Moreover, it is preferable that each site | part used for extraction is dried. For example, dry leaves and dry branches are preferable, and dry leaves are more preferable.

  Next, extraction of bamboo leaves and lemon myrtle will be described. In addition, a bamboo leaf and a lemon myrtle may be used for extraction operation separately, and the mixture of both leaves may be used for extraction operation.

  As described above, the extraction solvent used for extraction is preferably water, ethyl alcohol, or a water-ethyl alcohol mixture, and more preferably water or a water-ethyl alcohol mixture. When a water-ethyl alcohol mixture is used, the mixing ratio of the two is not particularly limited. For example, the mass ratio of water: ethyl alcohol is 10:90 to 90:10, or 20:80 to 80:20.

The extraction temperature is not particularly limited, and can be appropriately set from 0 ° C. to the boiling point of the solvent depending on the extraction solvent. More specifically, when the solvent is a water-ethyl alcohol mixture, the extraction temperature is preferably 30 ° C. to boiling point, more preferably about 60 to 80 ° C. When the solvent is water, the extraction temperature can be carried out at a temperature where the water is liquid (0 to 100 ° C.), preferably about 60 to 100 ° C., more preferably about 70 to 100 ° C. The extraction time is not particularly limited, and can be set as appropriate. For example, when the solvent is a water-ethyl alcohol mixture, it can be appropriately set according to the mixing ratio of ethyl alcohol and the extraction temperature. Preferably there is. When the solvent is water, the extraction time is about several tens of seconds to 24 hours, and preferably about 1 minute to 4 hours. The ratio of the extracted plant part (each part of the bamboo leaves and lemon myrtle used for extraction) and the extraction solvent is not particularly limited and can be set as appropriate. For example, the solvent is preferably 2 to 1000 parts by weight, more preferably 2 to 50 parts by weight, and still more preferably about 3 to 30 parts by weight with respect to 1 part by weight of the extracted plant part. The aqueous extract (aqueous extract) obtained by the extraction method as described above can be separated from the leaves using conventional methods such as filtration and centrifugation. The aqueous extract may be used as it is or after being concentrated (for example, concentrated to about Brix 50 to 70). Concentration is
For example, the reaction can be performed at about 30 to 50 ° C. under a reduced pressure of about 30 to 40 mmHg. Moreover, after concentration, solid content (for example, extract powder) can also be obtained by means such as freeze drying, spray drying, granulation drying, and the like. At that time, an excipient such as dextrin may be added. In this case, the powder may be used as it is as a leaf extract, or may be mixed and granulated with an optional component as necessary after producing the extract powder and used in the form of granules or fine granules.

  The contents of the koji leaf extract and the lemon myrtle extract in the composition according to the present invention are not particularly limited as long as the effects of the present invention are achieved. For example, it is preferable that 0.1-100 mass% is contained, and, it is more preferable that 1-99 mass% is contained in a camellia leaf extract or a persimmon leaf extract and a lemon myrtle extract in dry mass conversion. In addition, in the case where the koji leaf extract and the lemon myrtle extract are included, the content mass ratio (in terms of dry mass) of the koji leaf extract and the lemon myrtle extract is not particularly limited, but is preferably 1: 4 to 4: 1. 2: 3 to 3: 2 are more preferable.

In addition, the composition according to the present invention can be taken in one or a plurality of times (preferably 2 to 3 times). The subject of application is preferably a human, but may be a non-human mammal other than a human. Even when the subject of application is a non-human mammal (eg, a pet or livestock, more specifically, a dog, cat, cow, pig, chicken, sheep, etc.), the administration or intake amount of the human should be set as appropriate. Can do. Moreover, it is preferable that the application method to the living body of the composition according to the present invention is applied orally (that is, an oral composition).

  As described above, the composition according to the present invention can activate Tie2. When Tie2 is activated, the connection between endothelial cells / wall cells and endothelial cells is strengthened, and the structures of blood vessels and lymph vessels are stabilized. Thereby, various symptoms (problems) of the body can be improved. In particular, Tie2 decreases with aging and diseases, and is suitable for middle-aged and elderly people (for example, those over 30 years old, over 40 years old, over 50 years old, or over 60 years old) to improve various symptoms of the body. Such symptoms are not particularly limited, and include symptoms caused by unstable structures of blood vessels and lymph vessels, especially nutrients leaking from blood vessels, or waste products and water from lymph vessels. Symptoms caused by accumulation in the body without being collected for leakage are included. More specifically, for example, swelling of the body (especially hands, feet, face, etc.), cooling of the body (especially feet, hands, etc.), stiffness of the body (especially shoulders, waist, neck), skin (especially facial skin). ), Various symptoms such as wrinkles, sagging, and roughness are effective and effective in improving these symptoms. Therefore, the composition according to the present invention is intended for not only subjects who exhibit such symptoms, but also subjects who are expected to exhibit such symptoms in the future, and such symptoms. It is effective for a wide range of subjects such as those who want to avoid as much as possible. In particular, it is particularly suitable for middle-aged and older people (for example, 30 years old or older, 40 years old or older, 50 years old or older, or 60 years old or older), especially middle-aged women.

  The composition according to the present invention contains a koji leaf extract, or a kashiwa leaf extract and a lemon myrtle extract, and may further contain other components.

  Examples of other components include rosehip extract, hibiscus extract, dokudami extract, ginger extract, pearl barley extract, mugwort extract, chamomile (chamomile) extract, onion extract and the like. . As these extracts, those extracted from appropriate parts among the parts (leaves, stems, roots, fruits, etc.) of each plant can be used.

  The rosehip extract is preferably an extract from fruits, and more preferably an extract from dried fruits. The hibiscus extract is preferably an extract from straw and more preferably an extract from dry straw. The dokudami extract is preferably an extract from whole plants (eg, flowers, leaves, stems, roots, etc.), and more preferably an extract from a dried product. The ginger extract is preferably an extract from the root (tuberous root). The pearl barley extract is preferably an extract from fruit (seed), and more preferably an extract from dried fruit. The artemisia extract is preferably an extract from leaves and / or stems, and more preferably an extract from dry leaves and / or dry stems. The chamomile extract is preferably an extract from a flower, and more preferably an extract from a dried flower. The onion extract is preferably an extract from the edible leaf (the thick part of the leaf), and is an extract from the epidermis of the part (called the “onion skin”). It is more preferable. In addition, extraction can be performed on the same conditions as the extraction conditions of the above-mentioned bamboo leaf and lemon myrtle.

  As the other components, known components can be appropriately selected and used according to the field in which the composition is used. For example, a pharmaceutically or food hygienically acceptable carrier can be used.

  The composition according to the present invention can be preferably used as, for example, a pharmaceutical composition or a food composition (including a beverage composition and a food additive composition).

  When used as a pharmaceutical composition, other components include pharmaceutically acceptable bases, carriers, and / or additives (eg, solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders). , Disintegrants, lubricants, etc.). The form of the pharmaceutical composition is not particularly limited, and examples thereof include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, creams, and poultices. The pharmaceutical preparation of these forms can be prepared by a conventional method by combining the other ingredients with the koji leaf extract and / or the lemon myrtle extract as necessary.

  When used as a food composition, examples of other components include bases, carriers, additives, and other components and materials that can be used as food. Also, the form of the food composition is not particularly limited. For example, processed foods, health foods (dietary supplements, nutritional functional foods, foods for the sick, foods for specified health, functional indication products, etc.), supplements, sick people Foods for hospitals (hospital foods, sick foods, nursing foods, etc.) can be exemplified. These can be prepared by conventional methods. In particular, when preparing a food composition as a health food (dietary supplement, nutritional functional food, food for the sick, food for specified health use, functional labeling product, etc.) or supplement, continuous intake is easy to perform. Thus, for example, it is preferable to prepare in the form of granules, capsules, tablets (including chewable agents), beverages (beverage powders, drinks, etc.), among others, capsules, tablets, tablets, beverage powders, drinks, etc. Although the form is preferable from the viewpoint of easy intake, it is not particularly limited thereto. When used as a food additive composition among food compositions, examples of the form include liquid, powder, flake, granule, and paste. More specifically, seasonings (soy sauce, sauce, ketchup, dressing, etc.), flakes (sprinkles), grilled meat sauce, spices, roux paste (carrere paste, etc.) and the like can be exemplified.

  In addition, one preferable form of the composition according to the present invention may be tea.

  Moreover, this invention also includes the raw material for obtaining the said extract. For example, it also includes dried tea leaves or tea bags, including dried cocoon leaves and dried lemon myrtle. Furthermore, the present invention also includes the dried tea leaves or tea bags for improving the above-mentioned symptoms, including dried dried leaves or dried dried leaves.

  These dry tea leaves or tea bags may further contain other components. For example, dried rosehip berries, dried hibiscus pods, dried dokudami leaves, ginger root (preferably dried ginger roots), dried pearl barley (seed), dried mugwort leaves, dried chamomile flowers, onion skin (preferably dried onion Skin) etc. may be contained alone or in combination of two or more.

  These dry tea leaves or tea bags are preferable because water (including hot water) can be used to easily obtain tea included in the composition according to the present invention.

In addition, the content of each dry plant part in these dry tea leaves or tea bags is not particularly limited, but, for example, the dry bamboo leaves are preferably 20 to 40 parts by mass with respect to 100 parts by mass of the total dry plant part, 25-35 mass parts is more preferable. Moreover, 20-40 mass parts is preferable and, as for a dry lemon myrtle, 25-35 mass parts is more preferable. When dry rosehip fruit is included, 15 to 25 parts by mass is preferable, and 15 to 20 parts by mass is more preferable. When dry hibiscus koji is included, about 4 to 8 parts by mass is preferable. When dried dokudami leaves are included, about 2 to 6 parts by mass is preferable. When ginger root (preferably dry ginger root) is included, about 1 to 5 parts by mass is preferable. When dried pearl barley (seed) is included, about 1 to 5 parts by mass is preferable. When dry mugwort leaves are included, about 0.5 to 3 parts by mass is preferable. When a dry chamomile flower is included, about 0.5-3 mass parts is preferable. When onion skin is included, about 0.5 to 3 parts by mass is preferable.

In this specification, “including” includes “consisting essentially of” and “consisting of” (The term “comprising” includes “consisting essentially of” and “consisting of.”).

  Hereinafter, the present invention will be specifically described, but the present invention is not limited to the following examples. Hereinafter, “%” represents “% by mass” unless otherwise specified.

Examination of Tie2 activation Using HUVEC (human umbilical cord vascular endothelial cells), it was examined whether the Kashiwa leaf extract and the combination of Kashiwa leaf extract and lemon herb extract activate Tie2.

HUVECs were cultured in the growth medium described in Table 1 below (37 ° C., 5% CO ₂ ), and Tie2 activation was examined in a low serum medium.

  Each extract was prepared using machine-dried dried plants as follows. 4 g of each dried plant (mainly dried leaves) was weighed in a container, and 100 ml of boiling distilled water was added to the vessel and immersed for 5 minutes. The tea leaves were filtered using a tea bag filter, cooled to room temperature, and then filtered with a 0.22 μm pore size PES filter (Mirex GP, Millipore). The amount of soluble solid content was measured using a digital refractometer (RX-5000α, Atago Co., Ltd.). The solution was diluted with sterilized distilled water so that the soluble solid content was 0.5%. And the sample mother liquor was prepared with the combination of following (i)-(v) so that soluble solid content might be 0.5%. Then, 120 μL of the sample mother liquid and 1080 μL of the low serum medium were mixed, and 100 μL of the obtained mixed liquid was used as a sample-containing medium, and examination was performed using this.

  In addition, in the said examination, the herb extract which gives a lemon flavor was combined with the persimmon leaf extract.

<Combination>
Soluble solid content is (i) Kashiwa leaf extract 0.5%
(Ii) Persimmon leaf extract and lemon myrtle extract (LM), 0.375% + 0.125%, or 0.25% + 0.25%
(Iii) Persimmon leaf extract and lemongrass extract (LG), 0.375% + 0.125%, or 0.25% + 0.25%
(Iv) Persimmon leaf extract and lemon verbenaner extract (LV), 0.375% + 0.125%
Or 0.25% + 0.25%
(V) Persimmon leaf extract and lemon balm extract (LB), 0.375% + 0.125%, or 0.25% + 0.25%

In the examination, culture for 60 minutes in a sample-containing medium (37 ° C., 5% CO ₂ ), collect cells (HUVEC), and use a measurement kit Tie2 activation assay (Phospho / Total Tie-2 ELISA assay, R & D systems). Used to measure the activity of Tie2. As a positive control, angiopoietin-1 (Ang-1), which is a ligand of Tie2, was used instead of the measurement sample (concentration in medium: 1 μg / ml). PBS (phosphate buffered saline) was used as a negative control.

  The measurement results are shown in FIG. In FIG. 1, the Tie2 activation effect of each sample is shown as a relative value when the Tie2 activity in the negative control is 1. By the koji leaf extract alone, a Tie2 activation effect equivalent to that of Angiopoietin-1 was obtained. Moreover, when only lemon myrtle was combined with the koji leaf extract among each herb extract which gives lemon flavor, the outstanding Tie2 activation effect was shown.

  Furthermore, cell viability was also measured. Specifically, examination was performed using 100 μL of a low serum medium containing MTT under the above conditions. The MTT-containing low serum medium was prepared by adding DTT (3- (4,5-dimethyl-thiazol-2-yl) -2,5-diphenyltetrazolium bromide) powder to 5 mg / ml in D-PBS (Dulbeccolate buffered physiological medium). The solution is dissolved in a saline solution, filtered through a 0.22 μm pore size PES filter, and further diluted 10-fold with a low serum medium.

Cells (HUVEC) were cultured for 60 minutes in sample-containing medium (37 ° C., 5% CO ₂ ), the medium was removed, and further cultured overnight in MTT-containing low serum medium (37 ° C., 5% CO ₂ ). The medium was removed, 100 μL of DMSO was added, mixed for 5 minutes, and the absorbance at 570 nm wavelength was measured with a microplate reader. The results are shown in FIG. FIG. 2 shows the cell viability when each sample is used as a relative value when the cell viability (absorbance) in the negative control is 1. Compared with the koji leaf extract alone, the cell viability was higher when the herb extracts giving lemon flavor were used in combination, and the cell viability was highest when combined with lemon myrtle.

Preparation of Test Sample Various ingredients were mixed at the blending ratio shown in Table 2 below to prepare dry tea leaves. Each raw material is a dried product of each plant part mechanically dried. The tea leaves were put in a tea bag and used as a test sample (tea bag).

Test methods and results Questions from the “worker's fatigue accumulation self-diagnosis checklist” prepared by the Ministry of Health, Labor and Welfare for women in their 30s and 60s who have a habit of drinking tea. The subject was checked for subjective symptoms according to the items described. The subjective symptoms were answered about the physical condition for several days before the check. Then, the total score obtained was totaled, and 100 women with a total score of 9 or more were selected. The 100 people took tea prepared from a test sample (tea bag) once a day for 6 weeks. At this time, at the time of intake 0 weeks (immediately before intake) and 2 weeks, the following items were self-evaluated. In addition, at the time of 0 weeks of intake, check the “subject symptoms of the most recent 1 week” at that time, and at the time of 2 weeks of intake, check the “subject symptoms of the most recent 2-3 days” at that time. received.
・ Shoulders are stiff ・ Hands and feet are cold ・ Neck is stiff ・ Foot is easy to swell ・ Skin is dry ・ Skin is easy to be rough ・ It feels like there is no firmness ・ The face tends to be dull ・The face tends to sag and the face tends to swell Note that the self-evaluation was specifically performed in the following seven stages.
7). Very applicable 6. Applicable 5. Somewhat applicable 4. 2. I can't say either. Not so much 2. Not applicable 1. Does not apply at all

The results of the above self-assessment are shown in Table 3. In Table 3, “Previous” indicates the time of 0 week,
“After” indicates time points of 2 weeks, respectively. In any self-assessment item, it was confirmed that the symptoms were significantly improved.

  From the above, it was found that tea obtained from tea leaves mainly composed of persimmon leaves and lemon myrtle can activate Tie2, improve blood flow and lymph flow, and thus improve body symptoms. Moreover, from this, it was found that the koji leaf extract and the lemon myrtle extract have an excellent Tie2 activation effect.

Claims (10)

The composition for Tie2 activity containing a persimmon leaf extract. The composition according to claim 1, comprising a koji leaf extract and a lemon myrtle extract. The composition according to claim 1 or 2, which is an oral composition. The composition according to any one of claims 1 to 3, which is a pharmaceutical composition or a food composition. A method for producing a composition for Tie2 activity, comprising a step of subjecting persimmon leaves or persimmon leaves and lemon myrtle to extraction with water, ethanol, or a water-ethanol mixture. A dried tea leaf or tea bag comprising dried bamboo leaves and dried lemon myrtle. Dry tea leaves or tea bags for Tie2 activity, including persimmon leaves, persimmon leaves and lemon myrtle. The composition according to any one of claims 1 to 4, which is used for improving swelling, improving blood flow, improving lymph flow, improving body stiffness, improving cold, or improving wrinkles, sagging or roughness of skin. Including leaf extract, or leaf extract and lemon myrtle extract for swelling improvement, blood flow improvement, lymph flow improvement, body stiffness improvement, cold improvement, or wrinkle, sagging or roughening improvement ,Composition. Dried tea leaves, including dried koji leaves or dried koji leaves and dried lemon myrtle, for swelling improvement, blood flow improvement, lymph flow improvement, body stiffness improvement, cold improvement, or wrinkle, sagging or roughening improvement Or tea bags.