JP6147954B2 - Tie2 activator, vascular maturation agent, vascular normalization agent, and vascular stabilization agent - Google Patents

Description

  The present invention relates to a Tie2 activator, an angiogenesis inhibitor, a blood vessel maturation agent, a blood vessel normalizing agent, and a blood vessel stabilizer, and a pharmaceutical composition.

  A blood vessel has a structure in which vascular endothelial cells and vascular wall cells (vascular smooth muscle cells and perisite) adhere indirectly or directly via an extracellular matrix, and oxygen and nutrients are absorbed by living bodies. It has a function of supplying waste tissue and removing waste from living tissue.

  In general, the formation of blood vessels consists of vasculogenesis where new blood vessels are formed, and angiogenesis where new blood vessels are formed by the elongation and branching of existing blood vessels. And divided into two stages. In the former, vascular endothelial growth factor (VEGF) acts and is involved in a very wide range of angiogenesis from the initial development of blood vessels called angiogenesis to the subsequent angiogenesis, and the latter is angiopoietin. (Ang) acts and participates in the control of adhesion between vascular endothelial cells and vascular wall cells and the structural stabilization of blood vessels.

  Under normal oxygen conditions, blood vessel endothelial cells and blood vessel wall cells that line the periphery of blood vessels are firmly adhered to each other, and the blood vessel structure is kept stable. Cells can detach from vascular endothelial cells and proliferate disordered blood vessels. Such a phenomenon (angiogenesis) is often observed in diseases mainly composed of vascular lesions such as tumors, rheumatoid arthritis, diabetic retinopathy, hyperlipidemia, and hypertension.

  These angiogenesis are known to be suppressed by activating the receptor tyrosine kinase Tie2 (Tyrosine kinase with Ig and EGF homology domain 2) expressed in vascular endothelial cells (see, for example, Patent Document 1). ). In an ischemic disease caused by suppression of blood vessel narrowing or blood vessel enlargement, it has been reported that the blood vessel cavity is enlarged by activation of Tie2 (see, for example, Non-Patent Document 1). It has also been reported that the activation of Tie2 suppresses cell death of vascular endothelial cells (see, for example, Non-Patent Document 2).

Thus, it is known that the activation of Tie2 not only suppresses angiogenesis but also matures, normalizes, and stabilizes blood vessels.
For example, in vascular regenerative medicine, it is known that the activation of Tie2 induces adhesion between vascular endothelial cells and vascular wall cells in a blood vessel to mature the blood vessel.
For example, in diseases where vascular wall cells observed in tumors, diabetic retinopathy, etc. do not adhere to vascular endothelial cells, disordered blood vessels grow. By activation of Tie2, vascular wall cells become endothelial cells. It is known to adhere and normalize blood vessels.
For example, with respect to environmental factors that disrupt various intracellular and external blood vessel structures, it is known that activation of Tie2 suppresses instability of blood vessels and stabilizes blood vessels.

  As a naturally-occurring substance that suppresses angiogenesis by such activation of Tie2, cinnamon extract is known (for example, see Patent Document 1), but there is a problem that the activity is insufficient. Moreover, although suramin is known as a substance which suppresses angiogenesis (for example, refer patent document 2), there exists a problem that it is not excellent in safety.

  Accordingly, there is a strong demand for rapid development of highly safe substances having excellent Tie2 activation action, angiogenesis inhibition action, blood vessel maturation action, blood vessel normalization action, and blood vessel stabilization action. is the current situation.

JP 2009-263358 A JP-T 9-503488

Science. 1999 Dec 24; 286 (5449): 2511-4. P. N. A. S. 2004 Apr 13; 101 (15): 5553-8.

  An object of the present invention is to solve the conventional problems and achieve the following objects. That is, an object of the present invention is to provide a Tie2 activator having an excellent Tie2 activation action and high safety. Another object of the present invention is to provide a highly safe angiogenesis inhibitor having an excellent angiogenesis inhibitory action. In addition, the present invention has excellent blood vessel maturation action, blood vessel normalization action, and blood vessel stabilization action, and is a highly safe blood vessel maturation agent, blood vessel normalization agent, and blood vessel stabilization. An object is to provide an agent. The present invention also provides a highly safe pharmaceutical composition having excellent Tie2 activation action, angiogenesis inhibition action, blood vessel maturation action, blood vessel normalization action, and blood vessel stabilization action. For the purpose.

  As a result of intensive studies by the present inventors in order to solve the above problems, the following extract has excellent Tie2 activation action, angiogenesis inhibition action, blood vessel maturation action, blood vessel normalization action, and blood vessel stability. The present invention has been completed by knowing that it has a crystallization effect.

The present invention is based on the above findings by the present inventors, and means for solving the above problems are as follows. That is,
<1> Contains at least one extract of star fruit extract, ghetto extract, lotus extract, rooibos extract, Indian date extract, Karin extract, Syuju guava extract, and hihitsu extract as an active ingredient It is a Tie2 activator characterized by doing.
<2> Containing as an active ingredient at least one of star fruit extract, ghetto extract, lotus extract, rooibos extract, Indian date extract, Karin extract, Syuju guava extract, and hihatsu extract An angiogenesis inhibitor characterized by
<3> Containing as an active ingredient at least one of star fruit extract, ghetto extract, lotus extract, rooibos extract, Indian date extract, Karin extract, Syuju guava extract, and hihatsu extract A vascular maturation agent, a vascular normalization agent, or a vascular stabilization agent.
<4> The Tie2 activator according to <1>, the angiogenesis inhibitor according to <2>, and the blood vessel maturation agent, blood vessel normalizing agent or blood vessel stability according to <3> It is a pharmaceutical composition characterized by containing at least one of the agent.

According to the Tie2 activator of the present invention, it is possible to solve the conventional problems and to provide a highly safe Tie2 activator having an excellent Tie2 activation action.
According to the angiogenesis inhibitor of the present invention, the conventional problems can be solved, and an angiogenesis inhibitor having an excellent angiogenesis inhibitory action and high safety can be provided.
According to the blood vessel maturation agent, blood vessel normalization agent, and blood vessel stabilization agent of the present invention, the conventional problems are solved, and an excellent blood vessel maturation action, blood vessel normalization action, and blood vessel It is possible to provide a vascular maturation agent, a vascular normalization agent, and a vascular stabilization agent having a stabilizing action and high safety.
According to the pharmaceutical composition of the present invention, the above conventional problems are solved, and it has excellent Tie2 activation action, angiogenesis inhibition action, blood vessel maturation action, blood vessel normalization action, and blood vessel stabilization action. In addition, a highly safe pharmaceutical composition can be provided.

FIG. 1 is a diagram showing the results of detecting Tie2 phosphorylation in each sample by Western blotting. FIG. 2 is a diagram showing the results of detecting Tie2 phosphorylation in each sample by Western blotting. FIG. 3 is a diagram showing the results of detecting Tie2 phosphorylation in each sample by Western blotting. FIG. 4 is a diagram showing the results of detecting Tie2 phosphorylation in each sample by Western blotting. FIG. 5 is a diagram showing the results of detecting Tie2 phosphorylation in each sample by Western blotting.

(Tie2 activator, angiogenesis inhibitor, blood vessel maturation agent, blood vessel normalizing agent, blood vessel stabilizer, and pharmaceutical composition)
Tie2 activator, angiogenesis inhibitor, blood vessel maturation agent, blood vessel normalizing agent, blood vessel stabilizer, and pharmaceutical composition of the present invention are a star fruit extract, a ghetto extract, a lotus extract In addition, rooibos extract, Indian date extract, Karin extract, Syushu guava extract, and Hibatsu extract are contained as active ingredients, and further contain other ingredients as required.

  The Tie2 activator has a Tie2 activating action of phosphorylating Tie2 to convert it into its active form (phosphorylated Tie2). When Tie2 is activated, it induces autophosphorylation of intracellular tyrosine kinase domain and induces adhesion between vascular endothelial cells and vascular wall cells. In an ischemic disease caused by suppression of vascular narrowing or vasodilation, the vascular cavity is expanded by activation of Tie2. In addition, activation of Tie2 suppresses cell death of vascular endothelial cells.

  The angiogenesis inhibitor has an angiogenesis inhibitory action that suppresses a network of new blood vessels formed from existing blood vessels. In the hypoxic state, the activation of Tie2 is temporarily suppressed, the adhesion between the vascular endothelial cell and the vascular wall cell is dissociated, and a new blood vessel network is formed from the vascular endothelial cell from which the adhesion is dissociated. An angiogenesis inhibitor can suppress the disordered growth of blood vessels caused by the adhesion of such vascular wall cells to endothelial cells.

  The vascular maturation agent induces adhesion between vascular endothelial cells and vascular wall cells, and between vascular endothelial cells such that intravascular environmental factors (cells and humoral factors) do not easily leak out of the blood vessel. It has a maturation effect to form adhesion spots. In vascular regenerative medicine, activation of Tie2 can induce adhesion between vascular endothelial cells and vascular wall cells in the blood vessel, thereby allowing the blood vessel to mature.

  The blood vessel normalizing agent increases the adhesion between vascular endothelial cells and promotes the lining of the vascular wall cells to the vascular endothelial cells, thereby leading to disordered growth of blood vessels and blood vessels in which vascular permeability is broken. It has a normalizing action to make abnormal blood vessels normal. Also, in diseases where vascular wall cells observed in tumors, diabetic retinopathy, etc. do not adhere to vascular endothelial cells and proliferate disordered blood vessels, Tie2 activation activates vascular wall cells to become endothelial cells. It is possible to adhere and normalize the blood vessels.

  The above-mentioned blood vessel stabilizer has an action to suppress damage to existing blood vessels, dissociation between vascular endothelial cells, and dissociation between vascular endothelial cells and vascular wall cells, and stabilizing effect to suppress cell death of vascular endothelial cells. Have. In addition, with respect to environmental factors that break down various vascular structures inside and outside cells, activation of Tie2 can suppress vascular instability and stabilize blood vessels.

  The pharmaceutical composition is a composition comprising any of the Tie2 activator, the angiogenesis inhibitor, the vascular maturation agent, the vascular normalization agent, and the vascular stabilization agent, and Tie2 It has an activation effect, an angiogenesis inhibitory effect, a vascular maturation effect, a vascular normalization effect, and a vascular stabilization effect.

<Star fruit extract>
The star fruit is an evergreen tree of the genus Carambola, and the scientific name is Averhoa carambola (Japanese name: Carambola). The place of origin of the star fruit is not only in Southeast Asia but also in tropical to subtropical regions such as Southern China, Taiwan, Brazil, Guyana, Trinidad and Tobago, Florida, Hawaii, etc., which are easily available from these regions. The star fruit is classified into sweet berries that produce sweet berries and sour berries that produce sour berries, and is used for jams, jellies, and pickles in addition to raw food. The leaves of the star fruit are used as preventive or therapeutic agents for cold flu, acute gastroenteritis, urinary disadvantages, postpartum edema and the like.

  The extraction part of the star fruit is not particularly limited and can be appropriately selected according to the purpose.For example, a leaf part, a stem part, a flower part, a fruit part, a root part, and the like can be mentioned. Leaves are preferred.

  As a method for preparing the extraction part of the star fruit, it can be obtained by drying each part and then pulverizing it as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The star fruit extract can be easily obtained by a method generally used for plant extraction. There is no restriction | limiting in particular as said star fruit extract, According to the objective, it can select suitably, For example, the diluted solution of an extract, a concentrated liquid, the dried product of an extract, etc. are mentioned.

  The extraction method of the star fruit is not particularly limited and can be appropriately selected according to the purpose.For example, the extraction part of the star fruit as an extraction raw material is put into a treatment tank filled with an extraction solvent, Examples include a method of obtaining an extract by eluting soluble components while appropriately stirring as necessary and then filtering to remove the extraction residue.

  The extraction conditions (extraction time and extraction temperature) of the star fruit and the amount of the star fruit extraction solvent used are not particularly limited and may be appropriately selected depending on the purpose.

  There is no restriction | limiting in particular as an extraction solvent of the said star fruit, According to the objective, it can select suitably, For example, water, a hydrophilic solvent, or these mixed solvents are mentioned. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . In addition, it is preferable that the starfruit extraction solvent is used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a purification method of the said star fruit extract, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said star fruit extract, Although it can select suitably according to the objective, 100 microgram / mL-400 microgram / mL are preferable, and 200 microgram / mL-400 microgram / mL are more preferable.

<Getto extract>
The ghetto is an evergreen perennial plant belonging to the genus Gingeraceae, and its scientific name is Alpinia speciosa (also known as moon peach). The origin of the ghetto is from the southern end of Kyushu to Okinawa, the coast of Ogasawara, from Taiwan to southern China, Southeast Asia, India, etc., and is easily available from these regions. The height of the ghetto is 2m to 3m, the stems are upright, the leaves are paper-like, and they are oval skin needles in two rows, and the flowers are drooping large inflorescences. doing.

  There is no restriction | limiting in particular as an extraction site | part of the said ghetto, According to the objective, it can select suitably, For example, a leaf part, a stem part, a flower part, a fruit part, a root part etc. are mentioned, Among these, a leaf Part is preferred.

  As a method for preparing the extraction part of the ghetto, it can be obtained by drying each part and then pulverizing it as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The ghetto extract can be easily obtained by a method generally used for plant extraction. There is no restriction | limiting in particular as an extract of the said ghetto, According to the objective, it can select suitably, For example, the diluted solution of an extract, a concentrate, the dried product of an extract, etc. are mentioned.

  The extraction method of the ghetto is not particularly limited and can be appropriately selected according to the purpose.For example, the extraction site of the ghetto as an extraction raw material is put into a processing tank filled with an extraction solvent, and necessary. Accordingly, there may be mentioned a method of obtaining an extract by eluting soluble components while stirring appropriately and then filtering to remove the extraction residue.

  The extraction conditions (extraction time and extraction temperature) of the ghetto and the usage amount of the extraction solvent for the ghetto are not particularly limited and can be appropriately selected according to the purpose.

  There is no restriction | limiting in particular as an extraction solvent of the said ghetto, According to the objective, it can select suitably, For example, water, a hydrophilic solvent, or these mixed solvents are mentioned. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . The ghetto extract solvent is preferably used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a purification method of the said ghetto extract, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said ghetto extract, Although it can select suitably according to the objective, 50 microgram / mL-400 microgram / mL are preferable, and 200 microgram / mL-400 microgram / mL are more preferable.

<Lotus extract>
The lotus is a perennial aquatic plant belonging to the lotus family Lotus, and its scientific name is Nelumbo nucifera (alias: lotus). The lotus origin is in and around the Indian subcontinent and is readily available from these regions. The lotus extends a stem from an underground stalk in the ground and leaves a leaf on the surface of the water. The leaf has water repellency, is circular and has a petiole in the center, and has a plant height of about 1 m.

  The lotus extraction site is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include leaves, stems, embryos, flowers, fruits, roots, and the like. Among them, the embryo part is preferable.

  The lotus extraction part can be prepared by drying each part and then pulverizing the part as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The lotus extract can be easily obtained by a method generally used for plant extraction. There is no restriction | limiting in particular as said lotus extract, According to the objective, it can select suitably, For example, the dilution liquid of an extract, a concentrate, the dried product of an extract, etc. are mentioned.

  The lotus extraction method is not particularly limited and can be appropriately selected according to the purpose. For example, the lotus extraction site as an extraction raw material is put into a treatment tank filled with an extraction solvent, and is necessary. Accordingly, there may be mentioned a method of obtaining an extract by eluting soluble components while stirring appropriately and then filtering to remove the extraction residue.

  The lotus extraction conditions (extraction time and extraction temperature) and the amount of the lotus extraction solvent used are not particularly limited and may be appropriately selected depending on the purpose.

  The lotus extraction solvent is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include water, a hydrophilic solvent, and a mixed solvent thereof. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . The lotus extraction solvent is preferably used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a purification method of the said lotus extract, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said lotus extract, Although it can select suitably according to the objective, 100 microgram / mL-400 microgram / mL are preferable, and 200 microgram / mL-400 microgram / mL are more preferable.

<Rooibos extract>
The rooibos is a dicotyledonous plant belonging to the genus Asparatus , and its scientific name is Aspalathus linearis ( N. L. Barum .) R. It is Dahlgr (L eguminosae ) (plant name: Asparagus salinealis). The rooibos cultivated land is the South Africa's Cedar Burke Mountains and is readily available from these areas. The dried product of rooibos fermented young leaves and branches is said to be effective for antispasmodic drugs, tonics, vomiting, diarrhea, and other mild stomach upsets. The rooibos tea has a sweetness, does not contain caffeine, has a very low tannin concentration, and has an antioxidant effect.

  The rooibos extraction site is not particularly limited and can be appropriately selected depending on the purpose. Examples thereof include leaf, young leaf, branch, trunk, bark, flower, fruit, and root. Among these, young leaves and branches are preferable, and specifically, dry powder (rooibos tea) of young leaves and branches is preferable.

  As a method for preparing the rooibos extraction site, it can be obtained by drying each site and then pulverizing it as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The rooibos extract can be easily obtained by a method generally used for plant extraction. The rooibos extract is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include an extract diluted solution, a concentrated solution, and a dried extract.

  The rooibos extraction method is not particularly limited and can be appropriately selected according to the purpose. For example, the extraction portion of the rooibos as an extraction raw material is put into a treatment tank filled with an extraction solvent, and is necessary. Accordingly, there may be mentioned a method of obtaining an extract by eluting soluble components while stirring appropriately and then filtering to remove the extraction residue.

  The rooibos extraction conditions (extraction time and extraction temperature) and the amount of the rooibos extraction solvent used are not particularly limited and may be appropriately selected according to the purpose.

  The rooibos extraction solvent is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include water, a hydrophilic solvent, and a mixed solvent thereof. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . The rooibos extraction solvent is preferably used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a purification method of the said rooibos extract, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said rooibos extract, Although it can select suitably according to the objective, 50 micrograms / mL-400 micrograms / mL are preferable, and 200 micrograms / mL-400 micrograms / mL are more preferable.

<Indian Date Extract>
The Indian date is an evergreen tree plant belonging to the genus Tamarind of the genus Jacques, and the scientific name is Tamarindus India L. It is. The Indian dates are also called tamarind, grow naturally in tropical regions such as Asia and Africa, and are cultivated in regions such as India and Thailand, and are easily available from these regions. In Japan, the polysaccharide (tamarind seed gum) contained in the seed endosperm portion of Indian Date is used for foods such as thickeners, and tannin contained in the seed coat portion is used as a pigment.

The extraction site of the Indian dates is not particularly limited and can be appropriately selected according to the purpose.For example, leaves, branches, stems, bark parts, flower parts, fruit parts, seed parts, seed coat parts, Although a root etc. are mentioned, Among these, a seed coat part is preferable.
As a method for preparing the Indian dates extraction site, each site can be dried, and then can be obtained as it is or after being pulverized using a crusher and subjected to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The Indian dates extract can be easily obtained by a method generally used for plant extraction. The Indian Date extract is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a diluted solution of an extract, a concentrated solution, and a dried extract.

  The Indian date extraction method is not particularly limited and can be appropriately selected according to the purpose.For example, the extraction site of the Indian date as an extraction raw material is put into a treatment tank filled with an extraction solvent, Examples include a method of obtaining an extract by eluting soluble components while appropriately stirring as necessary and then filtering to remove the extraction residue.

  There are no particular limitations on the extraction conditions (extraction time and extraction temperature) of the Indian Date, and the amount of the Indian Date extraction solvent can be appropriately selected according to the purpose.

  The extraction solvent for Indian Date is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include water, a hydrophilic solvent, and a mixed solvent thereof. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . The Indian Date extraction solvent is preferably used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a purification method of the said Indian dates extract, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said Indian date extract, Although it can select suitably according to the objective, 10 microgram / mL-100 microgram / mL are preferable, and 10 microgram / mL-20 microgram / mL are more preferable.

<Karin extract>
The karin is a deciduous tree of the genus Rosaceae and the scientific name is Chaenomeles sinensis . Karin is native to eastern China and is cultivated in Japan, China, Korea, the United States, France, and the like, and is easily available from these regions. The fruit of the karin is called herb medicine (Wamokka), and is widely used as a cough to suppress inflammation from the old throat.

The extraction site of the karin is not particularly limited and may be appropriately selected depending on the purpose. Examples thereof include leaves, branches, trunks, bark parts, flower parts, fruit parts, etc. Of these, the fruit portion is preferred.
As a method for preparing the extraction part of the karin, it can be obtained by drying each part and then pulverizing it as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The karin extract can be easily obtained by a method generally used for plant extraction. There is no restriction | limiting in particular as an extract of the said karin, According to the objective, it can select suitably, For example, the diluted solution of an extract, a concentrate, the dried product of an extract, etc. are mentioned.

  The method for extracting the karin is not particularly limited and can be appropriately selected according to the purpose. For example, the extraction site of the karin as an extraction raw material is put into a treatment tank filled with an extraction solvent, and is necessary. Accordingly, there may be mentioned a method of obtaining an extract by eluting soluble components while stirring appropriately and then filtering to remove the extraction residue.

  The extraction conditions (extraction time and extraction temperature) of the karin and the amount of the extraction solvent used for the karin are not particularly limited and can be appropriately selected according to the purpose.

  There is no restriction | limiting in particular as the extraction solvent of the said karin, According to the objective, it can select suitably, For example, water, a hydrophilic solvent, or these mixed solvents are mentioned. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . Moreover, it is preferable to use the extraction solvent for the karin at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a purification method of the said karin extract, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said karin extract, Although it can select suitably according to the objective, 10 microgram / mL-400 microgram / mL are preferable, and 200 microgram / mL-400 microgram / mL are more preferable.

<Cidium guava extract>
The said guava is a shrub belonging to the genus Vanjiro, and the scientific name is Psidium guajava L. It is. The guaium is native to tropical America and is readily available from these regions. The above-mentioned guava is also called bunjirou and contains tannin, chlorophyll, folic acid, vitamins, minerals, proteins, polysaccharides and the like, which are polyphenol compounds, and is widely used as a health food.

There is no restriction | limiting in particular as an extraction site | part of the said guaium guava, According to the objective, it can select suitably, For example, a leaf part, a branch part, a trunk part, a bark part, a flower part, a fruit part, a root part etc. are mentioned. Of these, the fruit portion is preferred.
As a method for preparing the extraction site of the above-mentioned guaium guava, it can be obtained by drying each site and then pulverizing it as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The above-mentioned guaium guava extract can be easily obtained by a method generally used for plant extraction. There is no restriction | limiting in particular as an extract of the said guaium guava, According to the objective, it can select suitably, For example, the diluted solution of an extract, a concentrated liquid, the dried product of an extract, etc. are mentioned.

  The method for extracting the guaium guava is not particularly limited and can be appropriately selected according to the purpose.For example, the extraction site of the guaium guava as an extraction raw material is put into a treatment tank filled with the extraction solvent, Examples include a method of obtaining an extract by eluting soluble components while appropriately stirring as necessary and then filtering to remove the extraction residue.

  There is no restriction | limiting in particular as the extraction conditions (extraction time and extraction temperature) of the said guaium guava, and the usage-amount of the extraction solvent of the said guaium guava, According to the objective, it can select suitably.

  There is no restriction | limiting in particular as an extraction solvent of the said guaium guava, According to the objective, it can select suitably, For example, water, a hydrophilic solvent, or these mixed solvents are mentioned. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . Moreover, it is preferable to use the extraction solvent of the above-mentioned guaium guava at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a refinement | purification method of the extract of said guaium guava, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said guaium guava extract, Although it can select suitably according to the objective, 100 microgram / mL-800 microgram / mL are preferable, and 400 microgram / mL-800 microgram / mL are more preferable.

<Hihatsu extract>
The baboon is a vine evergreen tree of the family Pepperaceae, and its scientific name is Pipe longum . The Hibatsu is readily available from Southeast Asia. The jade of the cherries has a fleshy cylindrical shape, and the dried product is widely used as a spice.

The extraction site of the cherries is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a fruit head part, a leaf part, a stem part, a flower part, and a root part. Part is preferred.
As a method for preparing the extraction part of the chickpea, it can be obtained by drying each part and then pulverizing it as it is or using a crusher and subjecting it to solvent extraction. The drying may be performed in the sun, or may be performed using a commonly used dryer.

  The above extract of the cherries can be easily obtained by a method generally used for extraction of plants. There is no restriction | limiting in particular as an extract of the said cherries, According to the objective, it can select suitably, For example, the diluted solution of an extract, a concentrate, the dried product of an extract, etc. are mentioned.

  The method for extracting the honey is not particularly limited and can be appropriately selected according to the purpose. For example, the extraction part of the chick is extracted as a raw material for extraction into a treatment tank filled with the extraction solvent. Accordingly, there may be mentioned a method of obtaining an extract by eluting soluble components while stirring appropriately and then filtering to remove the extraction residue.

  There are no particular restrictions on the extraction conditions (extraction time and extraction temperature) of the chicks and the amount of the extraction solvent used for the chicks, and it can be appropriately selected according to the purpose.

  There is no restriction | limiting in particular as an extraction solvent of the said cherries, According to the objective, it can select suitably, For example, water, a hydrophilic solvent, or these mixed solvents are mentioned. The water is not particularly limited and may be appropriately selected according to the purpose.For example, pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, Examples include ion-exchanged water, physiological saline, phosphate buffer, and phosphate buffered saline. There is no restriction | limiting in particular as said hydrophilic solvent, According to the objective, it can select suitably, For example, C1-C5 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Acetone, methyl ethyl ketone, etc. Lower aliphatic ketones; C2-C5 polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin. There is no restriction | limiting in particular as said mixed solvent, Although it can select suitably according to the objective, When using a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, a lower aliphatic ketone. 1 to 40 parts by mass with respect to 10 parts by mass of water, and when using polyhydric alcohol, 1 to 90 parts by mass with respect to 10 parts by mass of water is preferably added. . In addition, it is preferable that the extraction solvent for Hibatsu is used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  There is no restriction | limiting in particular as a refinement | purification method of the extract of the said cherries, According to the objective, it can select suitably, For example, purification methods, such as liquid-liquid partition extraction, various chromatography, and membrane separation, are mentioned.

  There is no restriction | limiting in particular as a density | concentration of the said chickpea extract, Although it can select suitably according to the objective, 50 micrograms-400 micrograms / mL are preferable, and 200 micrograms / mL-400 micrograms / mL are more preferable.

<Other ingredients>
The other components are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include excipients, moisture-proofing agents, preservatives, reinforcing agents, thickeners, emulsifiers, antioxidants, and sweeteners. , Sour, seasoning, coloring, flavoring, whitening agent, moisturizer, oil component, UV absorber, surfactant, thickener, alcohol, powder component, colorant, aqueous component, water, skin nutrition Agents and the like.
The other components are not particularly limited and may be appropriately selected depending on the purpose. Specifically, hyaluronic acid, hyaluronic acid hydrolyzate, collagen, collagen hydrolyzate, ascorbic acid, ascorbine Acid derivatives, ascorbic acid glycosides, coenzyme Q10, propolis, royal jelly, royal jelly proteolysate, fucoidan, aloe powder, aloe extract, blueberry powder, blueberry extract, isoflavone, noni powder, noni extract, garlic powder, garlic Extract, turmeric powder, turmeric extract, chitosan, glucosamine, chlorella powder, chlorella extract, carnitine, maca powder, maca extract, cassis powder, cassis extract, hanabiratake powder, hanabiratake extract, and other beauty Plant powder and Or extract and the like.

<Application>
The Tie2 activator, the vascular maturation agent, the vascular normalization agent, the vascular stabilization agent, and the angiogenesis inhibitor, and the pharmaceutical composition of the present invention have excellent Tie2 activation action, angiogenesis inhibition action, Diseases mainly composed of vascular lesions such as tumors, rheumatoid arthritis, diabetic retinopathy, hyperlipidemia, hypertension, and atopy due to vascular maturation, vascular normalization, and vascular stabilization It can be suitably used as a pharmaceutical for allergic diseases such as dermatitis and hay fever, and as a safe preventive agent for these diseases, and its blending amount, usage, and dosage form are appropriately determined according to the purpose of use. You can choose. In addition, it was confirmed that the Tie2 activator, vascular maturation agent, vascular normalization agent, vascular stabilization agent, and angiogenesis inhibitor of the present invention are not digested in the digestive tract. Therefore, it can be suitably used as a food / beverage product such as a beauty food / beverage product or a health food / beverage product, and the blending amount, usage, and dosage form can be appropriately selected according to the purpose of use.

  The blending amount can be appropriately adjusted depending on the physiological activity of the extract, etc., but it can be adjusted as follows: Starfruit extract, Ghetto extract, Lotus extract, Rooibos extract, Indian date extract, Karin extract, Scidium guava extract In terms of at least one purified product of the product and the extract of Hihatsu, 0.0001% by mass to 20% by mass is preferable, and 0.0001% by mass to 10% by mass is more preferable.

  There is no restriction | limiting in particular as said usage, According to the objective, it can select suitably, For example, oral, parenteral, administration forms, such as external use, are mentioned.

  The dosage form is not particularly limited and may be appropriately selected depending on the intended purpose. For example, oral dosage forms such as tablets, powders, capsules, granules, extracts, and syrups, injections, and infusions And parenteral agents such as suppositories, ointments, creams, emulsions, lotions, packs, and external preparations such as bath preparations and hair cosmetics.

  Examples of the present invention will be described below, but the present invention is not limited to these examples.

[Test Example 1: Tie2 activation effect (Western blot) test]
(Example 1: Star fruit extract)
The pharmacological effect of the star fruit extract was evaluated by detecting the amount of phosphorylated Tie2 protein. The amount of the phosphorylated Tie2 protein was detected by an immunochemical technique using SDS-PAGE and Western blot. The procedure is shown below.
Tie2 phosphorylation analysis used cells obtained by overexpressing human Tie2 (Ba / F3-human Tie2) in mouse pro-B cells (Ba / F3). Stimulation of mouse pro-B cells was performed at 37 ° C. in a normal culture medium (10% FBS RPMI 1640 (manufactured by SIGMA, R-8755) +1 pg / mL mouse IL-3) with a star fruit extract (Maruzen Pharmaceutical Co., Ltd.). Manufactured, Star Fruit Leaf Extract Powder MF) (concentration unit: μg / mL) was added at a predetermined concentration (concentration shown in FIG. 1, concentration unit: μg / mL). After 15 minutes, the cells were washed with cold PBS, RIPA lysis buffer (50 mM Tris-HCl (pH 7.5); 150 mM NaCl; 1 mass% NP-40; 0.5 mass% sodium deoxycholate; 0.1 mass%) The cell extract was collected by SDS). The cell extract was electrophoresed on a 7.5% by mass SDS gel (NPU-7.5L, manufactured by Atto Co., Ltd.) and transferred to nylon membranes. The non-specific protein was blocked with 5 mass% skim milk / TBST for 60 minutes from the transcribed nylon membranes, and anti-Tie2 antibody (Ab33: Upstate), anti-phosphorylated-Tie2 antibody (Tyr992: Cell Signaling Technology) Blotted. Subsequently, blotting was performed with a secondary antibody labeled with HRP, and the obtained reaction product was photographed by visualizing a protein band with an electrochemiluminescence (ECL) solution. The results are shown in FIG.

(Example 2: Ghetto extract)
In Example 2, the star fruit extract was changed to a ghetto extract (manufactured by Maruzen Pharmaceutical Co., Ltd., moon peach leaf dry extract F), and the concentration shown in FIG. 1 (concentration unit: μg / mL) was used. Were tested in the same manner as in Example 1. The results are shown in FIG.

(Example 3: Lotus extract)
In Example 2, the star fruit extract was changed to a lotus extract (manufactured by Maruzen Pharmaceutical Co., Ltd., lotus germ extract powder MF), and the concentration shown in FIG. 1 (concentration unit: μg / mL) was used. Were tested in the same manner as in Example 1. The results are shown in FIG.

(Example 4: Rooibos extract)
In Example 2, the star fruit extract was changed to a rooibos extract (manufactured by Maruzen Pharmaceutical Co., Ltd., rooibos tea dry extract F), and the concentration shown in FIG. 2 (concentration unit: μg / mL) was used. Were tested in the same manner as in Example 1. The results are shown in FIG.

(Example 5: Indian dates extract)
In Example 2, the star fruit extract was changed to Indian Date Extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Indian Date Extract Powder MF), and the concentration shown in FIG. 3 (concentration unit: μg / mL) was used. The test was performed in the same manner as in Example 1 except for the above. The results are shown in FIG.

(Example 6: Karin extract)
In Example 2, the star fruit extract was changed to a karin extract (manufactured by Maruzen Pharmaceutical Co., Ltd., karin extract powder MF), and the concentration shown in FIG. 4 (concentration unit: μg / mL) was used. The test was conducted in the same manner as in Example 1. The results are shown in FIG.

(Example 7: Ciguaguava extract)
In Example 2, the above-mentioned star fruit extract was changed to a Jujuba guava extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Jujuba Guava dry extract F), and the concentration shown in FIG. 4 (concentration unit: μg / mL) was used. The test was performed in the same manner as in Example 1 except for the above. The results are shown in FIG.

(Example 8: Hihatsu extract)
The pharmacological effect of the chickpea extract was evaluated by detecting the amount of phosphorylated Tie2 protein. The amount of the phosphorylated Tie2 protein was detected by an immunochemical technique using SDS-PAGE and Western blot. The procedure is shown below.
Tie2 phosphorylation analysis used human umbilical vein endothelial cells (HUVECs). For stimulation of HUVEC by Ang1 and the subject, cells were first cultured in RPMI-1640 culture medium containing 1% FBS for 3 hours. Thereafter, the sample was added at the concentration shown in FIG. 5 (concentration unit: μg / mL), 20 minutes later, the cells were washed with cold PBS, RIPA lysis buffer (50 mM Tris-HCl (pH 7.5); 150 mM NaCl). 1% by mass NP-40; 0.5% by mass sodium deoxycholate; 0.1% by mass SDS). The cell extract was electrophoresed on a 7.5% by mass SDS gel and transferred to nylon membranes. The non-specific protein was blocked for 60 minutes with 5% by weight skim milk / TBST of the transcribed nylon membranes, and anti-Tie2 antibody (Ab33: Upstate), anti-phosphorylated-Tie2 antibody (Tyr992: Cell Signaling Technology) ). Subsequently, blotting was performed with a secondary antibody labeled with HRP, and the resulting reaction product was visualized and photographed with an electrochemiluminescence (ECL) solution. The concentration of the band was measured using Las 3000-mini's Multi Guage-Image software and calculated as p-Tie2 (each sample) / p-Tie2 (control). The results are shown in FIG.

(Reference Example 1: Positive control)
In Example 1, the above-mentioned star fruit extract was changed to Angiopoietin-1 (manufactured by R & D system) as a positive control, and the concentration (concentration unit: μg / mL) described in each drawing was used. Tested as in Example 1. The results are shown in each drawing.

(Reference Example 2: Negative control)
In Example 1, the test was conducted in the same manner as in Example 1 except that the star fruit extract was changed to only dimethyl sulfoxide (DMSO) as a negative control. The results are shown in each drawing.

[Test Example 1: Test results]
The test result of Tie2 activation action (Western blot) will be described.
1 to 5 show the phosphorylated Tie2 protein band in each sample detected by Western blotting. The intensity of the detected band is proportional to the amount of phosphorylated Tie2 protein. The darker the band, the greater the amount of phosphorylated Tie2 protein.
From FIG. 3 to FIG. 5, it was found that the Syushu guava extract, Indian Date extract, and Hibatsu extract induce Tie2 activation as much as Angiopoietin-1 which is a physiologically active substance. Among these, the Indian date extract was found to reach an activation level close to that of Angiopoietin-1 even at low concentrations. 1 and 2 and FIG. 4, the star fruit extract, ghetto extract, rooibos extract, lotus extract, and callin extract have weak Tie2 activation levels, but the induction of Tie2 activation was confirmed. It was.
In addition, Western blotting was performed using HUVEC for star fruit extract, ghetto extract, lotus extract, rooibos extract, Indian date extract, Karin extract, and Syushu guava extract. Similar results were obtained.
From the above, it was found that induction of activation of Tie2 was confirmed for the extracts used in the examples.

[Test Example 2: Tie2 activation action (immunoassay) test]
(Example 9: Star fruit extract)
Normal human umbilical vein endothelial cells (HUVEC) cultured to confluence were seeded in a 96-well plate at 2.0 × 10 ⁴ cells / 0.1 mL / well, and a medium for low serum vascular endothelial cell proliferation (Kurashiki Spinning) The culture was carried out overnight using Humedia-EG2). Next, the HUVEC after overnight culture was replaced with 0.1 mL of vascular endothelial cell basal medium (Humdia-EB2 manufactured by Kurashiki Boseki Co., Ltd.) 3 hours before cell stimulation (addition of test sample) and cultured again. Went. Thereafter, 0.1 mL of a star fruit extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Star Fruit Leaf Extract Powder MF) prepared as a test sample with the above-mentioned Humdia-EB2 at the concentration shown in Table 1 was added to the well. A minute incubation was performed. After the incubation, the amount of phosphorylated Tie2 and total Tie2 in the cells were measured according to the protocol using an immunoassay kit (manufactured by R & D Systems, Human Phospho-Tie2 (Y992) Immunoassay). The ratio was calculated.
For dimethyl sulfoxide (DMSO) used as a negative control, the ratio of phosphorylated Tie2 to total Tie2 was calculated in the same manner.
And Tie2 activation rate was calculated according to following formula (1), and phosphorylation action was evaluated. The results are shown in Table 1.
Tie2 activation rate (%) =
[{(Measured value of phosphorylated Tie2 when test sample is added) / (Measured value of total Tie2 when test sample is added)} / {(Measured value of phosphorylated Tie2 with negative control) / (With negative control) Measured value of total Tie2)}] × 100 (1)

(Example 10: Ghetto extract)
In Example 9, the star fruit extract was changed to a ghetto extract (manufactured by Maruzen Pharmaceutical Co., Ltd., moon peach leaf dry extract F), and the concentrations shown in Table 1 were used. And tested. The results are shown in Table 1.

(Example 11: Lotus extract)
In Example 9, the star fruit extract was changed to a lotus extract (manufactured by Maruzen Pharmaceutical Co., Ltd., lotus germ extract powder MF), and the concentrations shown in Table 1 were used. And tested. The results are shown in Table 1.

(Example 12: Rooibos extract)
In Example 9, the star fruit extract was changed to Rooibos extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Rooibos tea dry extract F), and the concentrations shown in Table 1 were used. And tested. The results are shown in Table 1.

(Example 13: Indian dates extract)
In Example 9, the star fruit extract was changed to Indian Date Extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Indian Date Extract Powder MF), and the concentrations shown in Table 1 were used. And tested. The results are shown in Table 1.

(Example 14: Karin extract)
In Example 9, the star fruit extract was changed to a Karin extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Karin Extract Powder MF), and the concentrations shown in Table 1 were used. Tested. The results are shown in Table 1.

(Example 15: Ciguaguava extract)
In Example 9, the above-mentioned star fruit extract was changed to a Shiju guava extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Shiju guava dried extract F), and the concentrations shown in Table 1 were used. And tested. The results are shown in Table 1.

(Example 16: Hihatsu extract)
In Example 9, the star fruit extract was changed to Hihatsu extract (manufactured by Maruzen Pharmaceutical Co., Ltd., Hihatsu Extract Powder MF), except that the concentrations shown in Table 1 were used. Tested. The results are shown in Table 1.

(Reference Example 3: Positive control)
In Example 9, the star fruit extract was tested in the same manner as in Example 9 except that the positive control was changed to Angiopoietin-1 (manufactured by R & D system) and the concentrations shown in Table 1 were used. . The results are shown in Table 1.

[Test Example 2: Test results]
The test result of Tie2 activation action (immunoassay) will be described.
When the Tie2 activation action was evaluated using the immunoassay kit, it was confirmed that Tie2 was phosphorylated and activated by the extract used in the examples. In the system to which DMSO as a negative control was added, no significant phosphorylation of Tie2 was observed. Further, in the system to which Angiopoietin-1 which is a positive control of Reference Example 3 was added, it was observed that Tie2 was phosphorylated and activated. Here, it is known that phosphorylation of Tie2 leads to vascular maturation, vascular normalization, and vascular stabilization, and angiogenesis is suppressed.
From the above, it was suggested that the extracts used in the Examples have a Tie2 phosphorylation effect, thereby suppressing angiogenesis and inducing vascular maturation, vascular normalization, and vascular stabilization. .

The Tie2 activator, angiogenesis inhibitor, blood vessel maturation agent, blood vessel normalizing agent, blood vessel stabilizer, and pharmaceutical composition of the present invention have excellent Tie2 activation action, angiogenesis inhibition action, Drugs related to diseases mainly composed of vascular lesions such as tumors, rheumatoid arthritis, diabetic retinopathy, hyperlipidemia, hypertension and the like because they have vascular maturation action, vascular normalization action, and vascular stabilization action It can be widely used as a safe preventive drug for these diseases.
It was also confirmed that the Tie2 activator, angiogenesis inhibitor, blood vessel maturation agent, blood vessel normalizing agent, and blood vessel stabilizer of the present invention are not digested in the digestive tract. Therefore, it can be widely used as food and drink such as beauty food and drink and health food and drink.

Claims (2)

A Tie2 activator characterized by containing an aqueous extract of starfruit leaves, a hydrophilic solvent extract, or a mixed solvent extract thereof as an active ingredient . Water extract of the leaves of star fruit, hydrophilic solvent extracts or maturation agent of the vessel, characterized in that it contains a mixture of these solvents extract as an active ingredient, normalizing agents of vascular or vascular stabilizing agent .