JP5926895B2 - Tie2 activator and lymphatic vessel stabilizer - Google Patents

Description

  The present invention includes a novel Tie2 activator comprising a plant of Mango Ginger (Curcuma amada ROXB.) Or an extract thereof and / or Panax Ginseng or an extract thereof, and the activator A lymphatic vessel stabilizer comprising:

  Blood is fed from the heart and returns to the heart via capillaries and veins. A lymphatic vessel forms a drainage path for tissue fluid separately from the vascular system. The lymphatic vessels maintain a closed circulatory system by keeping the blood volume constant by circulating interstitial fluid, proteins, fats, cells, and the like leaked from the blood vessels in the peripheral tissues to the vascular system. In the capillaries existing in the skin, the basement membrane surrounds the outside of the endothelial cells, and pericytes are further attached. On the other hand, in capillary lymphatic vessels, there is almost no basement membrane outside the endothelial cells, and pericytes are not attached. This structure is useful for efficiently taking in body fluids and cells from the interstitium (Non-Patent Document 1). So far, tyrosinase-type receptor VEGFR (vascular endothelial growth factor receptor) -3 has been shown to be specifically expressed in lymphatic endothelial cells, and its ligands VEGF-C and VEGF-D have been shown to form new lymph vessels. It was shown to induce Further, it has been clarified that VEGF-A induces lymphangiogenesis through VEGFR2 expressed in lymphatic endothelial cells (Non-patent Document 2). Furthermore, regarding the function of lymphatic vessels, there are the following reports. In mouse ears infected with adenovirus expressing VEGF-A, remarkable lymphangiogenesis was observed, but along with the structural abnormalities, lymphoid recovery function was also remarkable from experiments in which colloidal carbon was injected into the ear (Non-patent Document 3). That is, it is considered that lymphatic endothelial cells must be appropriately arranged and lined for the function of lymphatic vessels. We define this as “lymphatic vessel stabilization”.

  Physical or chemical stimulation to the skin induces angiogenesis and increased vascular permeability due to VEGF-A, etc., resulting in tissue fluid retention and edema. On the other hand, these stimuli are also known to directly induce lymphangiogenesis and expansion. So far, lymphatic dilation has been observed due to ultraviolet inflammation, and experiments with dye injection have revealed that lymphatic function is impaired. It is considered that lymphatic vessels are expanding and collecting interstitial fluid as water leaks into the dermis due to vasodilation. However, it was thought that excessive lymphatic vessel dilatation reduced the recovery function and delayed edema (Non-patent Document 4). That is, it is considered that “stabilization of lymphatic vessels” that does not induce excessive expansion of lymphatic vessels is necessary for rapid recovery of interstitial fluid.

  Up to now, secondary lymphedema associated with filaria, surgery, malignant tumor, and inflammation is known as a pathological condition involving dysfunction of lymphatic vessels as well as congenital lymphedema. Congenital lymphedema includes Milroy disease, Meige disease, and lymphedema-distichiasis syndrome. Milroy's disease has been reported to be aplastic and hypoplastic, while lymphedema-distichiasis syndrome has been reported to be hyperplastic. Also from these, it is considered necessary to maintain the recovery function not only by lymphatic neovascularization but also by stabilizing the lymphatic vessels (Non-patent Document 5).

Experimental Medicine Vol. 24, No. 18 (2006), pp. 133-138 Jussila L and Alitalo K, (2006) Vascular growth factors and lymphangiogenesis. Phisiol Rev 82: 673-700 Nagy et al., (2002) Vascular permeability factor / vascular endothelial growth factor induces lymphangiogenesis as well as angiogenesis.J Exp Med 196: 1497-1506 Kajiya K., Hirakawa S., and Detmar M., (2006) VEGF-A mediates UVB-induced impairment of lymphatic vessel function. Am J Pathol 169: 1496-1503 Experimental Medicine Vol. 24, No. 18 (2006), pp. 139-143 Kajiya K., et al., (2005) Hepatocyte growth factor promotes lymphatic vessel formation and function.EMBO J 24: 2885-95 Experimental Medicine Vol. 20, No.8 (2002), pp. 52-57

  An object of the present invention is to provide a novel Tie2 activator and, in turn, to provide a drug effective for stabilizing and improving the recovery function of lymphatic vessels by stabilizing the lymphatic vessels.

  Beginning with molecular cloning of VEGF, molecules of the VEGF family and angiopoietin (Angopoietin; Ang) family have been identified one after another as factors that act specifically on angiogenesis. VEGF and its receptors are involved in a very wide range of angiogenesis, from the initial development of blood vessels called angiogenesis to subsequent angiogenesis. On the other hand, Ang functions in luminal formation accompanied by cellular phenomena such as germination, branching, insertion, and retraction by vascular endothelial cells after angiogenesis. Ang is a receptor-type tyrosine kinase expressed in vascular endothelial cells, Tie (tyrosine kinase with Ig and EGF homology domain) -2, and vascular endothelial cells and vascular walls such as pericytes and vascular smooth muscle cells. It has been reported that it controls adhesion to cells and functions for structural stabilization of blood vessels (Non-patent Document 7).

In a previous study, the present inventor examined the function of Ang-1 by focusing on the relationship between Ang-1 and Tie2 expressed in lymphatic endothelial cells. As a result, Ang-1 was mediated through activation of Tie2. It has been clarified that the recovery function of lymphatic vessels is promoted (Japanese Patent Application No. 2010-517947). Furthermore, when the present inventors screened for a compound having Tie2 activation ability, a plant body of Mango Ginger (Curcuma amada ROXB.) Or an extract thereof, and / or Panax Ginseng or an extract thereof was found to be The present inventors have found that it has Tie2 activity like Ang-1, and has completed the following invention:
(1) A Tie2 activator comprising a plant body of mango ginger (Curcuma amada ROXB.) Or an extract thereof, and / or Panax Ginseng or an extract thereof.
(2) A lymphatic vessel stabilizer comprising the Tie2 activator.
(3) A cosmetic method for improving or preventing swelling, comprising applying the lymphatic vessel stabilizer of (2).

  By using the lymphatic vessel stabilizer according to the present invention, swelling and the like can be improved / prevented.

FIG. 1 is a graph showing changes in the amount of phosphorylated Tie2 in normal human umbilical vein endothelial cells (HUVEC) when mango ginger extract (Curcuma amada ROXB.) Extract or Panax Ginseng extract is added. is there.

Tie2 Activator In one aspect, the present invention provides a Tie2 activator comprising a plant of Mango Ginger (Curcuma amada ROXB.) Or an extract thereof, and / or Panax Ginseng or an extract thereof. provide.

  Activation of Tie2 refers to the ability to convert Tie2 into its active form (phosphorylated Tie2) by phosphorylation. As an activator of Tie2, it is well known that it has an activity to activate Tie2, such as Angiopoietin 1, or diphenhydramine or diphenhydramine hydrochloride in which the present inventors have found Tie2 activity, derived from plants belonging to the genus Cinnamomum There are extracts, or syringaresinol and 2-methoxycinnaldehyde.

The mango ginger (Curcuma amada ROXB.) Used in the present invention is a plant belonging to the genus Turmeric belonging to the ginger family and is an annual herb distributed or cultivated in the wild in India. Mango ginger (C. amada) can be used raw or dried, but it is preferably used as a dry powder or a solvent extract from the viewpoints of usability and formulation. Mango ginger can be used at any site such as whole grass, leaves, tuberous roots, flowers, seeds and the like. Of these, the tuberous root is preferably used.

  The mango ginger extract can be obtained by a conventional method. For example, after drying the mango ginger if necessary, it is immersed in an extraction solvent for a certain period of time or brought into contact with an extraction solvent that is heated to reflux, and then filtered. It can be obtained by concentrating. As the extraction solvent, any solvent that is usually used for extraction can be used. For example, water, methanol, ethanol, propylene glycol, 1,3-butylene glycol, glycerol and other alcohols, chloroform, dichloroethane, four Organic solvents such as carbon chloride, acetone and ethyl acetate can be used alone or in combination. The extract obtained by extraction with the above solvent is used as it is, or concentrated extracts are used, or these extracts are removed by using an adsorption method, for example, ion exchange resin, or a porous polymer (for example, Amberlite XAD- After adsorbing on the column of 2), elution with methanol or ethanol and concentration can also be used. Further, a partitioning method, for example, an extract extracted with water / ethyl acetate can be used.

  The mango ginger extract thus obtained is highly safe and has an excellent Tie2 activation action. With regard to mango ginger, JP 2010-222313 and the like have been known to promote hyaluronic acid production, but it has not been known at all that mango ginger and its extract have Tie2 activation activity. It has been confirmed for the first time by the inventor that these effects are obtained. The mango ginger extract is commercially available, for example, as “Mango Ginger Dry Extract F” (Maruzen Pharmaceutical Co., Ltd.), and these commercially available products may be used.

  Ginseng used in the present invention is widely known as (scientific name: Panax Ginseng), and its medicinal properties are known as a herbal medicine. Although it can be used raw or dried, it is preferably used as a dry powder or a solvent extract from the viewpoints of usability and formulation. As the extract, the root extract of ginseng (scientific name: Panax Ginseng) is most preferable, but the ginseng leaves, stems, branches, flowers, fruits, etc. also contain active ingredients. Any one or two or more extracts can also be used. Moreover, since the active ingredient similar to an extract is contained also in the pressing liquid obtained by pressing ginseng, the pressing liquid of ginseng can also be used instead of an extract.

  The method for extracting the active ingredient from ginseng is not particularly limited and is the same as in the case of mango ginger, and therefore detailed description thereof is omitted. According to one embodiment, ginseng root is soaked in, for example, water or an aqueous organic solvent and extracted at room temperature or from 80 ° C to 100 ° C. After filtering the extract obtained by the extraction treatment, it can be used as it is or after being concentrated or dried as necessary. In this extraction process, the ginseng root may be chopped or crushed. Further, raw roots or dry roots may be used, and roasted roots may be used. The roasting method is not particularly limited, and examples thereof include a method of roasting at 100 to 150 ° C. for 0.5 to 2 hours.

  The ginseng extract thus obtained is highly safe and has an excellent Tie2 activation action. For ginseng extract, for example, JP 2003-238365 has reported use as a hair-growth agent, but it has never been known that Ginseng and its extract have Tie2 activation action, It has been confirmed for the first time by the present inventor that these effects are obtained. The ginseng extract is marketed, for example as "Ginseng extract powder MF" (Maruzen Pharmaceutical Co., Ltd.) etc., and these commercial products may be used.

  In the present invention, the mango ginger plant or the extract thereof and the ginseng extract may be used alone or in combination. The amount of the mango ginger extract and / or ginseng extract in the Tie2 activator is 0.001 to 10% by mass, preferably 0.01 to 1.0% by mass, more preferably in the total amount of the agent. It is 0.1-1.0 mass% (dry mass).

Lymphatic vessel stabilizer In another aspect, the present invention provides a lymphatic vessel stabilizer comprising the Tie2 activator.

  The lymphatic vessel stabilizer according to the present invention can be used as a pharmaceutical or cosmetic effective for the treatment / prevention of various skin diseases such as edema (swelling) caused by leakage of lymph due to instability of the structure of lymphatic vessels. Examples of edema include secondary lymphedema associated with ultraviolet irradiation, filaria, surgery, malignancy, inflammation, and congenital lymphedema, such as Milroy disease, Meige disease, and lymphedema-distichiasis syndrome.

  The dose, usage, and dosage form of the lymphatic vessel stabilizer according to the present invention can be appropriately determined according to the purpose of use. For example, the dosage form of the lymphatic vessel stabilizer of the present invention is not particularly limited, and may be oral, parenteral, external use, etc., but is preferably an external preparation. Examples of the dosage form include external preparations such as ointments, creams, emulsions, lotions, packs, bath preparations, parenteral preparations such as injections, drops, or suppositories, or tablets, powders, capsules, granules, Oral administration agents such as extract and syrup can be mentioned.

  The blending amount of the Tie2 activator in the lymphatic vessel stabilizer of the present invention can be appropriately determined according to the use, but generally 0.0001-20.0 mol% can be blended in the total amount of the agent, preferably 0.0001-10.0 mol%, More preferably, it is 0.1-1 mol%.

  In addition to the Tie2 activator, the lymphatic vessel stabilizer of the present invention includes, for example, excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, and the like used in ordinary foods and pharmaceuticals. Antioxidants, sweeteners, acidulants, seasonings, colorants, fragrances, whitening agents, moisturizers, oily ingredients, UV absorbers, surfactants, thickeners, alcohols, powders commonly used in cosmetics, etc. Components, colorants, aqueous components, water, various skin nutrients, and the like can be appropriately blended as necessary.

  Further, when the lymphatic vessel stabilizer of the present invention is used as an external preparation for skin, auxiliary agents commonly used for external preparations for skin, such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate Sequestering agents such as gluconic acid, caffeine, tannin, verapamil, tranexamic acid and its derivatives, licorice extract, grabrizine, hot water extract of karin fruit, various herbal medicines, tocopherol acetate, glycyrrhizic acid and its derivatives or their derivatives Drugs such as salts, whitening agents such as vitamin C, magnesium ascorbate phosphate, glucoside ascorbate, arbutin, kojic acid, sugars such as glucose, fructose, mannose, sucrose, trehalose, retinoic acid, retinol, retinol acetate, palmiticin Acid retinol It can also be of vitamin A as appropriate formulation.

Cosmetic Method In yet another aspect, the present invention provides a cosmetic method for improving or preventing swelling, comprising applying a lymphatic vessel stabilizer.

  The cosmetic method according to the present invention is intended to reduce or prevent swelling and eye bags. This cosmetic method is applied to, for example, a site with swelling of the lymphatic vessel stabilizer according to the present invention and left as it is, or subjected to, for example, massage according to the direction of the flow of the lymphatic vessel. For example, the liquid flow can be promoted. This part can be applied to all parts of the body such as the face, neck, limbs, and the like.

  Next, the present invention will be described in more detail with reference to examples. In addition, this invention is not limited by this.

Western blotting of vascular endothelial cells
Normal human umbilical vein endothelial cells (HUVEC) expressing Tie2 were purchased from Sanko Junyaku. After culturing HUVEC in EBM-2 (Cambrex; Verviers, Belgium) to which additional factors such as growth factors were added, each mango ginger extract with a concentration mass of 0.1% (mango ginger dry extract F, Maruzen Pharmaceutical Co., Ltd.) )) And ginseng extract (Ginseng extract powder MF, manufactured by Maruzen Pharmaceutical Co., Ltd.), proteins in HUVEC were extracted with Phosphosafe Extraction Reagent (Novagen, Madison, WI). As a control, HUVEC with DMSO added was also prepared.

  The total protein amount was quantified with RC DC Protein Assay Kit (BIO-RAD, Hercules, CA) and detected by Western blotting as follows. SDS-PAGE was performed on 7.5% acrylamide gel (NPU-7.5L, ATTO, Japan) with an equal amount of total protein, and the expression of Tie2 and phosphorylated Tie2 protein was confirmed by antibody (Santa Cruz Biotechnology, Santa Cruz, The color was developed with ECL Kit.

  From the results shown in FIG. 1, it can be seen that each mango ginger extract and ginseng extract show a remarkable Tie2 activation action as compared with control DMSO. Since Ang-1 having the ability to activate Tie2 promotes the recovery function of lymphatic vessels, it is considered that these drugs also have the effect of stabilizing lymphatic vessels in the same manner as Ang-1.

Claims (1)

Tie2 (tyrosine kinase with Ig and EGF homology domain-2) activator comprising a plant body of mango ginger (Curcuma amada ROXB.) Or its extract and / or Panax Ginseng or its extract ( Except when not used as a Tie2 activator but for the purpose of improving swelling or prevention) .

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