WO2012101746A1

WO2012101746A1 - Tie-2 activator, agent for maturation, normalization, or stabilization of blood vessels, lymph vessel stabilizing agent, wrinkle prevention/improvement agent, and edema improvement/prevention agent

Info

Publication number: WO2012101746A1
Application number: PCT/JP2011/051245
Authority: WO
Inventors: 大田　正弘
Original assignee: 株式会社資生堂
Priority date: 2011-01-24
Filing date: 2011-01-24
Publication date: 2012-08-02

Abstract

The present invention provides: a Tie-2 activator comprising one or more types of compounds selected from a group consisting of Eleutheroside E, Eleutheroside E1, Sesamin, Eudesmine, Sylvatesmin, Pinoresinol, Yangambin, Forsythinol, and Coumarine; an agent for maturation, normalization, or stabilization of blood vessels and a lymph vessel stabilizing agent that contain the activator; and a wrinkle prevention/improvement agent and edema improvement/prevention agent that are characterized by being a combination of the agents.

Description

Tie2 activator, vascular maturation, normalization or stabilization agent, lymphatic vessel stabilizer, wrinkle prevention / improving agent and swelling improvement / prevention agent

The present invention includes Eleutheroside E, Eleutheroside E1, Sesamin, Eudesmine, Sylvatesmin, Pinoresinol, Yangambin, Forsythinol, and Coumarin. A novel Tie2 (Tyrosine Kinase with Ig and EGF Homology Domain 2) activator (Tie2 phosphorylating agent) consisting of one or more compounds selected from the group consisting of, vascular maturation, normalization or stabilization, Provided are a lymphatic stabilizer, a wrinkle prevention / improvement agent, and a swelling improvement / prevention agent.

In blood vessels, vascular smooth muscle cells and pericytes collectively called vascular wall cells are adhered to vascular endothelial cells via the extracellular matrix or directly from the outer cavity surface of vascular endothelial cells. The ratio of adhesion between endothelial cells and wall cells differs depending on the size of the blood vessel diameter. In the large vascular system, one layer of endothelial cells is lined with multiple layers of wall cells. In many cases, one wall cell is lined with one cell, and in a blood vessel having a smaller diameter, one wall cell adheres to a plurality of endothelial cells. Thus, the lining of wall cells against vascular endothelial cells is important for the structural “maturation” process of blood vessels. In addition, adhesion between vascular endothelial cells can form adhesion spots between endothelial cells so that intravascular environmental factors (cells and humoral factors) do not easily leak out of the blood vessel. It is important for the “maturation” process. Furthermore, depending on the demand for oxygen and nutrients in the tissue, especially when they are deficient, the blood vessels increase blood flow by expanding the lumen so that oxygen and nutrients can be adequately distributed to the tissue. Adjust. In other words, the process of controlling the permeability by controlling adhesion by forming adhesion spots between vascular endothelial cells, structurally stabilizing vascular endothelial cells with wall cell lining, and controlling the vascular cavity to large, medium and small , Defined as “blood vessel maturation”.

Also, in various pathological conditions, blood vessels with disordered structures are constructed, and immature blood vessels in which adhesion between endothelial cells is suppressed or adhesion of wall cells to endothelial cells is formed. These cause disordered increase in vascular permeability, thereby causing abnormal humoral factors and cellular traffic in the tissues and blood vessels. Increased permeability causes tissue edema, causes tissue dysfunction, and causes inflamed inflammatory cell infiltration, resulting in inflammation. Furthermore, since mural cells suppress sprouting of blood vessels from existing blood vessels, sprouting of blood vessels becomes excessive from blood vessels not accompanied by mural cells, and disordered sprouting of blood vessels is induced, resulting in worsening of the disease state. Such a phenomenon is observed in pathological conditions represented by diabetic retinopathy, tumor, and inflammation. In other words, blood vessels that have broken vascular permeability and abnormal blood vessels that cause disordered growth of blood vessels are promoted by lining the endothelial cells to the endothelial cells by increasing adhesion between the endothelial cells, thereby making the blood vessels normal. The state is defined as “normalization of blood vessels”. In addition, abnormal blood vessels such as those described above may cause changes in the internal and external blood flow factors, such as diabetes, hyperlipidemia, and hypertension, which may cause damage (cell death, etc.) to endothelial cells and mural cells, as well as cancer and inflammation. This triggers an excessive increase in the production of angiogenesis-promoting factors. When such a pathological condition occurs, it is defined as “blood vessel stabilization” to suppress damage to existing blood vessels, suppress dissociation of endothelial cells, or suppress dissociation of endothelial cells and wall cells. This stabilization also includes a mechanism for suppressing cell death of endothelial cells.

Angiogenesis is a phenomenon in which a new blood vessel network is formed from existing blood vessels, and is deeply related to diseases mainly consisting of vascular lesions such as tumors, rheumatoid arthritis, diabetic retinopathy, hyperlipidemia, and hypertension. Yes. Beginning with the molecular cloning of VEGF (vascular endothelial growth factor), VEGF family and angiopoietin (Angiopoietin; Ang) family molecules have been identified one after another as factors that act specifically on angiogenesis. It was. 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 regulates adhesion between vascular endothelial cells and vascular wall cells such as pericytes (pericytes) and vascular smooth muscle cells via the receptor tyrosine kinase Tie-2 expressed in vascular endothelial cells. It functions in structural stabilization (Non-patent Document 1: Experimental Medicine Vol.20, No.8-8 (2002) pp.52-57).

So far, four isoforms from Ang-1 to 4 are known, Ang-1 and Ang-2 are present in both humans and mice, but Ang-3 is present in mice and Ang-4 is present in humans. . Ang-1, -4 secreted from mural cells stimulates Tie-2 and induces autophosphorylation of intracellular tyrosine kinase domain, integrin activation, focal adhesion kinase (focal adhesion kinase; FAK) activation Involves activation of PI3K / Akt (phosphatidylinositol-3-kinase; PI3K, serine-threonine kinase: Akt) and induces adhesion between endothelial cells and mural cells. In normoxia, endothelial cells maintain their adhesion between endothelial cells and mural cells by Ang-1, -4, which is constantly secreted by mural cells, but when hypoxia occurs locally, Ang-1 , -4 antagonist Ang-2, -3 production is increased, Tie2 activation is temporarily suppressed, and adhesion between endothelial cells and wall cells lining it is suppressed. Endothelial cells proliferate by the dissociation of mural cells and initiate sprouting angiogenesis, leading to the formation of new vascular networks. Tie2 activation induces adhesion between endothelial cells and mural cells, thereby contributing to the stabilization of the vascular structure, and promotes adhesion between endothelial cells to control vascular permeability. In addition, Tie2 activation is also known to suppress cell death of endothelial cells (Non-patent Document 2: Cho CH, Kammerer RA, Lee HJ, Yasunaga K, Kim KT, Choi HH, Kim W , Kim SH, Park SK, Lee GM, Koh GY. Designed angiopoietin-1 variant, COMP-Ang1, protects against radiation-induced endothelial cell apoptosis. Proc Natl Acad Sci U S A. 2004 A15 -8), with respect to environmental factors disrupting the vascular structures of various intracellular and extracellular, by inducing activation of Tie2, thereby suppressing the instability of the vessel, it is stabilized and normalize blood vessel Is possible. Further, in vascular regenerative medicine, in the vessels built by vascular endothelial cells, by inducing activation of Tie2, it is possible to induce the adhesion of endothelial cells and parietal cells maturation of blood vessels. In addition, in diseases such as tumors and diabetic retinopathy, where disordered blood vessels grow due to the inability of mural cells to adhere to endothelial cells, activation of Tie2 causes mural cells to adhere to endothelial cells, It is possible to normalize blood vessels. According to the literature (Non-Patent Document 3: Thurston G, Suri C, Smith K, McClain J, Sato TN, Yancopoulos GD, McDonald DM. Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin 286 (5449): 2511-4), Tie2 activation has been reported to enlarge the vascular lumen, and in ischemic diseases caused by vascular narrowing or suppression of vasodilation By activating Tie2, it is possible to enlarge the vascular cavity and improve the disease state.

Recently, a plurality of molecules having coiled-coiled and fibrinogen-like domains, which are structural features of Ang, have been discovered. None of these have binding ability to Tie-1 receptor and Tie-2 receptor, so they are regarded as a group of molecules different from the existing Ang family and named Angiopoietin-like protein (Angptl). Angptl-1, -2, -3, -4, -5, -6, -7 have been reported. All Angptl receptors are orphan ligands whose receptors have not been identified at the present time, but they are also expected to exhibit various actions.

In other than vascular endothelial cells, activation of Tie2 is known to induce cell dormancy. According to previous reports, it has been reported that activation of Tie2 on hematopoietic stem cells induces dormancy of hematopoietic stem cells (Non-Patent Document 4: Arai F, Hirao A, Ohmura M, Sato H, Matsuoka S, Takubo K, Ito K, Koh GY, Suda T. Tie2 / angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell. 2004 Jul 23; 118 (2): 149-61). That is, by inducing activation of Tie2, it is possible to maintain the long-term survival of hematopoietic stem cells in vitro. In addition, according to previous reports, it is known that activation of Tie2 induces cell adhesion to an extracellular matrix or the like by activation of an adhesion factor such as integrin (Non-patent Document 5: Takakura). N, Huang XL, Naruse T, Hamaguchi I, Dumont DJ, Yancopoulos GD, Suda T. Critical role of the TIE2 endothelial cell receptor in the development of definitive hematopoiesis. Immunity6.77 By inducing cell adhesion, hematopoietic stem cells can be induced to maintain anchorage-dependent survival in vivo and externally by activation of Tie2. Furthermore, recent reports have suggested that Tie2 is expressed in cancer stem cells that are considered to be the most malignant in cancer tissues and are involved in cancer recurrence (non- Patent Document 6: Lee OH, Xu J, Fueyo J, Fuller GN, Aldape KD, Alonso MM, Piao Y, Liu TJ, Lang FF, Bekele BN, Gomez-Manzano C. Expression of the receptor Trosie is associated with integrin beta1-dependent adhesion to the extracellular matrix. Mol Cancer Res. 2006 Dec; 4 (12): 915-26). Just as activation of Tie2 can put the cell cycle of hematopoietic stem cells into a dormant state, activation of Tie2 expressed in cancer stem cells can also suppress the growth of cancer stem cells. .

The lymphatic vessels form 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 capturing body fluid or cells from efficiently stroma (Non-Patent Document 7:.. Experimental Medicine Vol 24, No. 18 (2006), pp 133-138). So far, it has been shown that the tyrosinase receptor VEGFR (vascular endothelial growth cept factor receptor) -3 is specifically expressed in lymphatic endothelial cells, and its ligands VEGF-C and VEGF-D are vascularized It was shown to induce In addition, VEGF-A was found to induce lymphangiogenesis through VEGFR2 expressed in lymphatic endothelial cells (Non-patent Document 8: Jussila L and Alitalo K, (2006) Vascular growth factors and lymphangiogenesis. Phisiol Rev 82: 673-700). 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 9: Nagy et al., (2002) Vascular permeability factor / vascular endothelial growth factor induces lymphangiogenesis as well as angiogenesis. J Exp Med 196: 1497-1506). 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 vascular permeability due to VEGF-A, etc., resulting in tissue fluid accumulation 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, excessive lymphatic dilation was thought to reverse the recovery function and delay edema (Non-Patent Document 10: Kajiya K., Hirakawa S., and Detmar M., (2006) VEGF -A mediates UVB-induced impairment of lymphatic vessel function. Am J Pathol 169: 1496-1503). 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.

So far, pathological conditions involving dysfunction of lymphatic vessels are known to include congenital lymphedema, as well as filaria, surgery, malignant tumors, and secondary lymphedema associated with inflammation. 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 11: Experimental Medicine Vol. 24, No. 18 (2006), pp. 139-143).

The object of the present invention is to provide a novel Tie2 activator, and in turn to maintain and enhance the recovery function of the lymphatic vessel by stabilizing the vascular maturation, normalization, stabilizing agent and lymphatic vessel To provide an effective medicine to do.

A detailed study of the behavior in the skin tissue irradiated with ultraviolet B revealed that adhesion of endothelial cells and wall cells was suppressed by ultraviolet irradiation, and Tie2 phosphorylation (activation) was suppressed. And, from experiments that examined the action of various herbal extracts on cells that forcedly expressed Tie2, cinnamon extract derived from plants belonging to the genus Cinnamomum had an action to activate Tie2. Has been found (International Publication No. WO2009 / 123211). Further, in the same patent publication, as a result of applying a cinnamon extract to mouse skin that has been photoaged by irradiation with ultraviolet B, the improvement effect is specifically shown by applying the extract. Thus, the extract endothelial cells by Tie2 phosphorylation - improving the adhesion between the parietal cells, promote, to form a stable vascular structure, to recover the UVB-induced skin damage and wrinkle formation by extension Has been confirmed. Therefore, it has been clarified that Tie2 activation contributes to vascular maturation, normalization or stabilization.

On the other hand, as a result of investigating the function of Ang-1 by lymph drainage assay focusing on the relationship between Ang-1 and Tie2 expressed in lymphatic endothelial cells, the relationship between Tie2 and lymphatic vessels It has been clarified that the recovery function of lymphatic vessels is promoted through the activation of PCT (International Application No. PCT / JP2009 / 061047). Therefore, it has been clarified that Tie2 activation contributes not only to vascular maturation, normalization or stabilization but also to lymphatic vessel stabilization and promotes lymph flow.

When the present inventor screened for various drugs having Tie2 activation ability, among the drugs examined, eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, Yangan bin, forcytinol, and coumarin are Tie2 It has been found to have activity and has led to the completion of the following invention:
(1) A Tie2 activator comprising one or a plurality of compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin.
(2) Maturation, normalization, or stabilization of blood vessels composed of one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin. .
(3) A wrinkle prevention / improving agent comprising the blood vessel maturation, normalization or stabilization agent of (2).
(4) A cosmetic method for preventing or improving wrinkles, comprising applying to the subject the vascular maturation, normalization or stabilizing agent of (2).
(5) A lymphatic stabilizer comprising one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin.
(6) A swelling improving / preventing agent comprising the lymphatic vessel stabilizer of (5).
(7) A cosmetic method for improving or preventing swelling, comprising applying a lymphatic vessel stabilizer of (5) to a subject.
(8) A method for maturation, normalization, or stabilization of blood vessels, wherein a subject in need of maturation, normalization, or stabilization of blood vessels is treated with eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol Applying a Tie2 activator consisting of one or more compounds selected from the group consisting of: Yanggambin, forcitinol and coumarin.
(9) A method for lymphatic vessel stabilization, comprising a subject in need of lymphatic vessel stabilization, eleuteroside E, eleuteroside E1, sesamin, eudesmin, silbatesmin, pinoresinol, Yangan bin, forcytinol and coumarin. Applying a Tie2 activator comprising one or more compounds selected from the group.
(10) One or more kinds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin for vascular maturation, normalization or stabilization Use of a Tie2 activator comprising a compound.
(11) Tie2 activity consisting of one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin for the stabilization of lymphatic vessels Use of chemicals.

According to the present invention, eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcitinol and / or coumarin activates Tie2, whereby the blood vessels are matured, normalized or stabilized, and The lymphatic vessels are stabilized and / or wrinkles and swelling are improved as a result.

It is a Western blot figure which shows the phosphorylation of Tie2 in a normal human umbilical vein vascular endothelial cell (HUVEC) at the time of adding an eleuteroside E, an eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol, or coumarin.

Tie2 Activator In one aspect, the present invention relates to Tie2 activity comprising one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin. Providing an agent.

Eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol are components contained in cereal and sesame seed lipids, and are known to have antioxidative and cholesterol lowering effects. The chemical structural formulas of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin and forcytinol are shown below.

Coumarin is well known as an aromatic compound derived from legumes and cereals. The chemical structural formula of coumarin is shown below.

All of these compounds are known to have effects such as Tie2 activation, vascular maturation, normalization or stabilization associated with Tie2 activation, or lymphatic vessel stabilization, as well as improvement of wrinkles and swelling. It is not done.

When the optical isomer is present, the compound may be optically pure or a mixture of optical isomers. In addition, the compound may be an inorganic salt or an organic salt. The salt is not particularly limited, and examples of the inorganic salt include hydrochloride, sulfate, phosphate, hydrobromide, sodium salt, potassium salt, magnesium salt, calcium salt, ammonium salt and the like. . Organic salts include acetate, lactate, maleate, fumarate, tartrate, citrate, methanesulfonate, p-toluenesulfonate, triethanolamine salt, diethanolamine salt, amino acid salt, etc. Can be mentioned.

Activating Tie2 refers to the ability to convert Tie2 into its active form (phosphorylated Tie2) by phosphorylating it. As an activator of Tie2, angiopoietin 1 (Ang-1) has been known to activate Tie2.

Eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin can be used alone or in combination as a Tie2 activator. Moreover, since Tie2 activation action is shown in a concentration-dependent manner, the compounding amounts of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin are included in the total amount of the composition formulated as an agent. 0.1 ppM to 100 mM, preferably 1 ppM to 100 mM, 10 ppM to 10 mM.

Vascular maturation, normalization or stabilizing agent In another aspect, the present invention relates to one or more selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin. A vascular maturation, normalization or stabilization agent comprising a plurality of compounds is provided.

The vascular maturation, normalization or stabilization agent according to the present invention can be used as a pharmaceutical or cosmetic effective for prevention and improvement of various diseases and aging caused by vascular structural changes. The vascular maturation, normalization or stabilization agent of the present invention induces normalization and stabilization of blood vessels, thereby causing various inflammatory diseases, immune diseases, adult diseases, etc., for example, various infectious diseases. It can be used as a drug for improving lesions throughout the body accompanied by neovascularization or vascular rupture in rheumatoid arthritis, gout, hypertension, diabetes, arteriosclerosis, atopic dermatitis, etc. In addition, by suppressing vascular permeability, edema in organs, organs and various tissues including the skin, such as inflammatory and immune diseases, vascular edema due to adult diseases, UV exposure, insect bites, allergies, etc. It can be used as a pharmaceutical or cosmetic to improve symptoms such as edema and itching caused by hypersensitivity. Furthermore, Tie2 phosphorylating agent is a cell death of endothelial cells induced by various factors, for example, inflammatory diseases, immune diseases, adult diseases, such as various infectious diseases, cancer, rheumatoid arthritis, gout, hypertension, It can be used as a pharmaceutical or a cosmetic that can suppress the destabilization of blood vessels by suppressing the death of endothelial cells due to diabetes, arteriosclerosis, radiation damage, various drugs or ultraviolet rays. The vascular maturation, normalization, or stabilizing agent of the present invention promotes wound healing, such as trauma or itch, by vascular maturation or enlargement of the vascular cavity, vascular maturation in revascularization medicine, ischemic It can be used in pharmaceuticals as an ameliorating agent for diseases such as cerebral infarction and myocardial infarction, and can also be used as an internal medicine and external medicine for low back pain, frostbite, alopecia, etc. by utilizing the effect of improving blood flow. In increasing cerebral blood flow, application to dementia is also possible. It can also be used as a cosmetic for alopecia. In cancer, it can be used as a therapeutic agent for inducing dormancy of cancer cells, and in stem cells as a maintenance agent in vivo and in vivo.

Photoaging refers to changes in skin appearance and function that are generally recognized as a result of repeated exposure to sunlight. Ultraviolet rays (UV), which is a component of sunlight, particularly intermediate UV (wavelength 290-320 nm, called UVB) mainly cause photoaging. The amount of UVB exposure necessary to cause photoaging is currently unknown. However, repeated exposure to UVB at levels that cause erythema and sunburn usually leads to photoaging. Clinically, photoaging is rough skin, wrinkle formation, spot coloration, soil coloration, sagging formation, onset of telangiectasia, development of mole, onset of purpura, vulnerable, atrophy, fibrosis It can be specified as the occurrence of a pigment removal region, the development of a pre-malignant tumor, a malignant tumor, and the like. Photoaging usually occurs in skin that is habitually exposed to sunlight, such as the face, ears, head, neck, and hands. In skin aging, photoaging due to skin damage or exposure to ultraviolet rays is the main cause, but this Tie2 phosphorylating agent can be used to improve photoaging by suppressing vascular damage that causes photoaging.

Lymphatic vessel stabilizer In another aspect, the present invention comprises one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin. Provide lymphatic vessel stabilizers.

The lymphatic vessel stabilizer according to the present invention can be used as a pharmaceutical or a cosmetic effective for the treatment / prevention of various skin diseases such as edema (swelling) due to leakage of lymph fluid caused by 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 Tie2 activator, vascular maturation, normalization or stabilization agent, and lymphatic vessel stabilizer according to the present invention can be appropriately determined according to the intended use. For example, the administration form of the vascular maturation, normalization or stabilizer and lymphatic vessel stabilizer of the present invention is not particularly limited, and may be oral, parenteral, external, etc., but preferably external It is an agent. 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 extracts and syrups can be mentioned, and can also be blended in foods such as tablets, drinks and cookies.

Selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin in the vascular maturation, normalization or stabilization and lymphatic vessel stabilizers of the present invention The compounding amount of one or more kinds of compounds can be appropriately determined according to the use, but is generally 0.1 ppM to 100 mM, preferably 1 ppM to 100 mM, 10 ppM to 10 mM in the total amount of the composition to be compounded.

In addition, the vascular maturation, normalization or stabilization agent and lymphatic vessel stabilization agent of the present invention may be selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yanganbin, forcytinol and coumarin. In addition to the selected compound or compounds, for example, excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidity used in normal foods and pharmaceuticals Whitening agent, moisturizer, oily component, ultraviolet absorber, surfactant, thickener, alcohols, powder component, colorant, aqueous component, which are usually used in cosmetics, seasonings, colorants, fragrances, etc. Water, various skin nutrients, and the like can be appropriately blended as necessary.

Furthermore, when the vascular maturation, normalization or stabilization agent or lymphatic vessel stabilization agent of the present invention is used as a skin external preparation such as a wrinkle prevention / amelioration agent or swelling improvement / prevention agent, Conventional auxiliaries, e.g. disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, sequestering agents, caffeine, tannin, verapamil, tranexamic acid and its derivatives, licorice Extracts, hot water extract of glabrizine, karin fruit, various herbal medicines, drugs such as tocopherol acetate, glycyrrhizic acid and its derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, glucoside ascorbate, arbutin, kojic acid, etc. Whitening agent, glucose, fructose, mannose, , Sugars trehalose, retinoic acid, retinol, retinol acetate, can also be suitably incorporated, such as vitamin A such as retinol palmitate. The amount of vascular maturation, normalization or stabilizer or lymphatic vessel stabilizer to be added to the external preparation for skin can be determined as appropriate, but generally 0.1ppM to the total amount of the composition to be added 100 mM, preferably 1 ppM to 100 mM, 10 ppM to 10 mM.

Cosmetic Method In another aspect, the present invention provides a cosmetic method for preventing and improving wrinkles, comprising applying a vascular maturation, normalization or stabilizing agent to a subject. The cosmetic method according to the present invention can be applied to facial skin where fine lines are likely to be formed, particularly skin such as the corners of the eyes, the lower eyelid, or the mouth, but is not limited to these parts.

In still another aspect, the present invention provides a cosmetic method for improving or preventing swelling, comprising applying a lymphatic vessel stabilizer to a subject. 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, proteins in HUVEC were expressed in the presence of each compound shown in the following table using Phosphosafe Extraction Reagent (Novagen, Madison, WI ). An HUVEC supplemented with Angiopoietin-1 (R & D system, 923-AN) as a positive control and DMSO as a negative control was also prepared.

Total protein the RC DC Protein Assay Kit (BIO-RAD, Hercules, CA) and quantified by and detected by Western blotting as follows. SDS-PAGE was performed on 7.5% acrylamide gel (NPU-7.5L, ATTO, Japan) with the same 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. The results are shown in FIG.

From the results in FIG. 1, compared to control DMSO, eleuteroside E, eleuteroside E1, sesamin, eudesmin, (+)-silvathesmin, (+)-pinoresinol, Yangan bin, forcytinol and coumarin are the same as Ang-1. It can be seen that it exhibits a remarkable Tie2 activation action. Therefore, it was suggested that these extracts all exhibited Tie2 phosphorylation effect and contributed to vascular maturation, normalization and stabilization, and as a result, angiogenesis could be suppressed. Furthermore, since Ang-1 having the ability to activate Tie2 promotes the recovery function of lymphatic vessels, it is considered that these extracts also have the effect of stabilizing lymphatic vessels in the same manner as Ang-1.

Formulation Example of Tie2 Activator Formulation examples of tablets, soft capsules, granules, drinks, candy, and cookies containing the Tie2 activator according to the present invention are shown below. These formulation examples are listed for the purpose of illustration, and are not intended to limit the technical scope of the present invention.

Formulation Example 1 (tablet)
(Composition) Formulation amount (mg / tablet)
Tie2 activator of the present invention 360.5
Lactose 102.4
Carboxymethylcellulose calcium 29.9
Hydroxypropylcellulose 6.8
Magnesium stearate 5.2
Crystalline cellulose 10.2
　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　
515.0

Formulation Example 2 (tablet)
(Composition) Formulation amount (mg / tablet)
Sucrose ester 70
Crystalline cellulose 74
Methylcellulose 36
Glycerin 25
Tie2 activator of the present invention 475
N-acetylglucosamine 200
Hyaluronic acid 150
Vitamin E 30
Vitamin B6 20
Vitamin B2 10
α-Lipoic acid 20
Coenzyme Q10 40
Ceramide (konjac extract) 50
L-proline 300
　　　　　　　　　　　　　　　　　　　　　　　　　　　
1500

Formulation Example 3 (soft capsule)
(Composition) Blending amount (mg / 1 capsule)
Edible soybean oil 530
Eucommia extract 50
Carrot extract 50
Tie2 activator of the present invention 100
Royal Jelly 50
Maca 30
GABA 30
Beeslow 60
Gelatin 375
Glycerin 120
Glycerin fatty acid ester 105
　　　　　　　　　　　　　　　　　　　　　　　　　　
1500

Formulation Example 4 (soft capsule)
(Composition) Blending amount (mg / 1 capsule)
Brown rice germ oil 659
Tie2 activator of the present invention 500
Resveratrol 1
Lotus germ extract 100
Elastin 180
DNA 30
Folic acid 30
　　　　　　　　　　　　　　　　　　　　　　　　　　
1500

Formulation Example 5 (granule)
(Composition) Blending amount (mg / pack)
Tie2 activator of the present invention 400
Vitamin C 100
Soy isoflavone 250
Reduced lactose 300
Soybean oligosaccharide 36
Erythritol 36
Dextrin 30
Fragrance 24
Citric acid 24
　　　　　　　　　　　　　　　　　　　　　　　　　
1200

Formulation Example 6 (Drink)
(Composition) Compounding amount (in g / 60 mL)
Eucommia extract 1.6
Carrot extract 1.6
Tie2 activator of the present invention 1.6
Reduced maltose starch syrup 28
Erythritol 8
Citric acid 2
Fragrance 1.3
N-acetylglucosamine 1
Hyaluronic acid Na 0.5
Vitamin E 0.3
Vitamin B6 0.2
Vitamin B2 0.1
α-Lipoic acid 0.2
Coenzyme Q10 1.2
Ceramide (konjac extract) 0.4
L-proline 2
Purified water residue
　　　　　　　　　　　　　　　　　　　　　　　　　　　
60

Formulation Example 7 (candy)
(Composition) Compounding amount (% by weight)
Sugar 50
Minamata 48
Tie2 activator of the present invention 1
Fragrance 1
　　　　　　　　　　　　　　　　　　　　　　　
100

Formulation Example 8 (Cookie)
(Composition) Compounding amount (% by weight)
Soft flour 45.0
Butter 17.5
Granulated sugar 20.0
Tie2 activator of the present invention 4.0
Egg 12.5
Fragrance 1.0
　　　　　　　　　　　　　　　　　　　　　　　　　　
100.0

Production Method of Formulation Example 8 (Cookie) Granulated sugar was gradually added while stirring the butter, and the egg, the Tie2 activator of the present invention (eleuteroside E) and the fragrance were added and stirred. After thorough mixing, a weak flour that was shaken uniformly was added and stirred at a low speed. Then, it shape | molded and baked for 170 degreeC15 minutes, and was set as the cookie.

Claims

The group consisting of Eleutheroside E, Eleutheroside E1, Sesamin, Eudesmine, Sylvatesmin, Pinoresinol, Yangambin, Forsythinol and Coumarine A Tie2 activator comprising one or more compounds.
Vascular maturation, normalization or stabilization comprising one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin.
A wrinkle prevention / improving agent comprising the blood vessel maturation, normalization or stabilization agent according to claim 2.
A cosmetic method for preventing or improving wrinkles, comprising applying to the subject the vascular maturation, normalization or stabilization agent according to claim 2.
A stabilizer for lymphatic vessels comprising one or more compounds selected from the group consisting of eleuteroside E, eleuteroside E1, sesamin, eudesmin, silvathesmin, pinoresinol, yangambin, forcytinol and coumarin.
A swelling improving / preventing agent comprising the lymphatic vessel stabilizer according to claim 5.
A cosmetic method for improving or preventing swelling, comprising applying the lymphatic vessel stabilizer according to claim 5 to a subject.