JP2010090036A - Vascularization inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To solve problems such as a problem wherein conventional vascularization inhibiting action-having substances are effective for preventing or treating vascularization diseases but not always guarantee safety for human bodies on the point of side effects, and a problem wherein conventional food-originated substances do not have sufficient vascularization-inhibiting actions, thereby further requiring new food-originated substances. <P>SOLUTION: There is provided the vascularization inhibitor containing carnosic acid, preferably only carnosic acid, as an active ingredient. The vascularization inhibitor can be administered or inoculated for a long period, because the vascularization inhibitor has higher safety for human bodies and can be produced at a lower production cost than those of conventional vascularization inhibitors. Thereby, the vascularization inhibitor overcomes conventional problems, has extremely high safety for human bodies, and can be provided at a low cost. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

The present invention relates to an angiogenesis inhibitor containing carnosic acid as an active ingredient for preventing and treating angiogenesis-related diseases.

Carnosic acid, an evergreen shrub native to the Mediterranean coast and extracted from the herb "Rosmarinus of ficinalis" and "Sage (Salvia officinalis)" has an important role in the maintenance of neurons It has been reported that it has the effect of enhancing the production of nerve growth factor that fulfills Furthermore, it has also been confirmed that administration of carnosic acid can prevent necrosis of brain tissue due to cerebral infarction and promote neurite outgrowth, and brain dysfunction such as Alzheimer-type dementia and Parkinson's disease Expected to be effective for diseases (Patent Document 1, Patent Document 2, Non-Patent Document 1). Therefore, many researches have been conducted on the effect of maintaining and growing nerve cells of rosemary and sage extracts, especially carnosic acid.

  On the other hand, despite many studies on carnosic acid, there has been no literature that reports that rosemary and sage extracts, especially carnosic acid, have angiogenesis-inhibiting activity. This is because the angiogenesis-inhibiting action, which is the effect of suppressing the growth of vascular endothelial cells, and the above-mentioned effect of maintaining and growing the nerve cells are completely opposite effects, so it is difficult to find an angiogenesis-inhibiting effect. It shows that there was.

  In addition, it has been reported that PCA (peracetilated carnosic acid), which is a derivative of carnosic acid, has an angiogenesis-inhibiting action (Non-patent Document 2). There are many examples where are completely different. That is, the effect obtained with the derivative is not always obtained with the basic substance. Incidentally, since PCA is reported to have a small amount of cytotoxicity at the same time, it is difficult to administer to the human body as an angiogenesis inhibitor.

  Rosemary and sage have long been used as a Western herb for a wide range of uses such as medicinal, fragrant, and cooking, and are known to be highly safe plants for the human body. Furthermore, rosemary extract containing carnosic acid as a main component has already been sold as a dietary supplement food. Therefore, safety to the human body at the time of oral administration of carnosic acid has been confirmed.

  By the way, the above-mentioned angiogenesis phenomenon is a physiological phenomenon in which a new blood vessel branch is made from an existing blood vessel and a blood vessel network is constructed. This phenomenon plays an important role in the female sexual cycle and the wound healing process of the skin, and is a physiological phenomenon necessary for the living body, but it is also important for the progression of many diseases called angiogenic diseases. Playing a role. In addition to tumor or cancer growth / metastasis, angiogenic diseases include diabetic retinopathy and age-related macular degeneration, which are causes of blindness, as well as chronic inflammatory diseases such as psoriasis and rheumatoid arthritis.

  In the above diseases, angiogenesis is a phenomenon necessary for supplying nutrients necessary for the growth and metastasis of a diseased tissue. Conversely, it is considered that the above-mentioned diseases can be prevented or treated by suppressing angiogenesis.

  Moreover, although it is in a research stage, it has been reported that angiogenesis and wrinkle formation (Non-patent Document 3), angiogenesis and obesity (Non-patent Document 4) are related to diseases other than diseases. It has been suggested that the agent is effective not only for angiogenic diseases but also for the prevention of beauty and metabolic syndrome.

Various angiogenic and inhibitory factors are known in angiogenesis. Promoters include vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), cytokines such as tumor necrosis factor (TNF-α) and interleukin-8 (IL-8), matrix metalloproteinase (MMP), etc. And other proteases. On the other hand, suppressors include cytokines such as interferon α / β (IFN-α / β), interleukin-12 (IL-12), transforming growth factor-β (TGF-β), and interferon inducible protein- And proteins such as 10 (IP-10) and plasminogen activator inhibitor (PAI).

  The angiogenesis process proceeds due to the complex action of the above-described angiogenesis-promoting factors and inhibitory factors. The angiogenesis process consists of the following four steps. 1) Angiogenesis is switched on by angiogenesis-promoting factors produced from cancer and stroma 2) Basement membrane under vascular endothelial cells is digested by involvement of MMP and plasminogen activator 3) Vascular endothelium Cells migrate and proliferate. 4) Vascular endothelial cells form lumens.

  Usually, the suppressor is more strongly expressed than the promoter, but it is known that the expression of the promoter exceeds the expression of the suppressor in the case of angiogenic diseases such as cancer. Therefore, an angiogenesis inhibitory factor is administered to a living body to suppress angiogenesis as a therapeutic method for angiogenic diseases. In that case, as an angiogenesis inhibitory function, it is considered that any one or a plurality of the above four steps may be suppressed.

  As angiogenesis inhibitors, to date, angiostatin (Non-patent Documents 5 and 6), endostatin (Non-patent Document 7), fumagillin derived from Aspergillus fumigatus and synthetic derivatives thereof are available. Certain TNP-470 (Non-patent Document 8), Cytogenin (Non-patent Document 9), Batimastat (BB-94) and Marimastat (BB-2516) (Non-patent Documents 10 and 11) which are metalloprotease inhibitors Synthetic chemical substances as well as monoclonal antibodies that inhibit the binding of angiogenic factors (EGF, TGF-α, VEGF, etc.) and their corresponding receptors are known (Non-patent Document 12). However, these substances have a drawback that they need to be considered for side effects and cannot be used easily.

Thus, in order to solve the problem of side effects, attempts have been made to search for substances having an anti-angiogenic action from food-derived substances. For example, shark cartilage (Non-patent Document 13), epigallocatechin (EGC) or epigallocatechin gallate (EGCG) (Non-patent Document 14), which is a green tea component, genistein (Non-patent Document 15 and 16) and the like have been reported to have an anti-angiogenic effect. Furthermore, it has been reported that a mixture of lycopene and carnosic acid has an anti-angiogenic effect (Patent Document 3). However, in this document, lycopene is the main substance and carnosic acid can be replaced by many other substances, so that carnosic acid itself has an angiogenesis inhibitor. is not.
Takumi Satoh, et.al. Carnosic acid protects neuronal HT22 Cells through activation of the antioxidant-resposive element in free carboxylic acid- and catechol hydroxyl moieties-dependent manners., Neuroscience Letters, 2008, 434, p260-265 Bertl E, et al., Inhibition of endothelial cell functions by novel potential cancer chmopreventive agents., Biochemical and Biophysical Research Communications, 2004, 325, p287-295 K.Yano, et al. Targeted overexpression of the angiogenesisinhibitor thrombospondin-1 in the epidermis of transgenic mice prevents ultraviolet-B-induced angiogenesis and cutaneous photo-damage., THE JOURNAL OF INVESTIGATIVE DERMATOLOGY, 2002,118 (5), p800- Five. MARupnick, et al. Adipose tissue mass can be regulatedthrough the vasculature., Proc. Natl. Acad. Sci. USA., 2002, 99 (16), p10730-5. MS O'Reilly, et al., Angiostatin induces and sustains dormancy of human primary tumors in mice., Nature Medicine, 1996, 2, p689-692 BK Sim, et al., A recombinant human angiostatin protein inhibits experimental primary and metastatic cancer, Cancer Reseach., 1997, 57, p1329-1334 MS O'Reilly, et al., Endostatin: an inherent inhibitorof angiogenesis and tumor growth., Cell, 1997, 88, p277-285 D. Ingber, et al., Synthetic analogues of fumagillin thatinhibit angiogenesis and suppress tumour growth., Nature, 1990, 348, p555-557 T. Oikawa, et al., Effects of cytogenin, a novel microbial product, on embryonic and tumor cell-induced angiogenic responses in vivo., Anticancer Reseach, 1997, 17, p1881-1886 G. Taraboletti, et al., Inhibition of Angiogenesis and Murine Hemangioma Growth by Batimastat, a Synthetic Inhibitor of Matrix Metalloproteinases, Journal of National Cancer Institute., 1995, 87, p293-298 Marimastat: BB 2516, TA 2516., Drugs in R & D, 2003,4 (3), p198-203. S.Iqbal and HJLenz, Integration of novel agents in the treatment of colorectal cancer., HYPERLINK "http://www.ingentaconnect.com/content/klu/280" \ o "Cancer Chemotherapy and Pharmacology" Cancer Chemotherapy and Pharmacology, 2004 , 54 Suppl.1, pS32-39 Y.AKIKUNI, Evaluation of the Effectiveness of Fork-Medicine Like Foods.Role of Angiogenesis Inhibitor in Novel Immunotherapy for Cancer (NITC)., Biotherapy, 2000,14 (10), p973-982 Y.Cao and R.Cao, Angiogenesis inhibited by drinking tea., Nature, 1999,398, p381 T. Fotsis, et al., Genistein, a Dietary-Derived Inhibitor of in vitro Angiogenesis., Proc. Natl. Acad. Sci. USA, 1993, 90, p2690-2694 Kento Miura et al., Product illustration Development of natural antitumor substance GCP, New Food Industry, 2001, 43 (3), p17-22 JP 2007-230945 A JP 2007-230946 A JP 2005-526719 A

The above-mentioned compounds having an anti-angiogenic action are considered to be effective for the prevention or treatment of angiogenic diseases. However, in view of side effects, there is a problem that safety to the human body is not necessarily guaranteed. .

  Moreover, it cannot be said that the conventional food-derived substance has sufficient anti-angiogenic action, and the price is high for long-term ingestion, and further food-derived substances are required. The present invention has been made in view of such circumstances, and an object of the present invention is to find an active ingredient having an excellent anti-angiogenic action and provide it as an anti-angiogenic agent.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have found a novel anti-angiogenic action for carnosic acid and have completed the present invention.

  That is, the present invention relates to an angiogenesis inhibitor containing carnosic acid as an active ingredient or an angiogenesis inhibitor containing only carnosic acid as an active ingredient.

  Furthermore, the present invention is a method for treating a disease caused by angiogenesis, particularly a malignant tumor, using the above-mentioned angiogenesis inhibitor, and particularly characterized in suppressing migration or proliferation of vascular endothelial cells. It relates to a treatment method. Furthermore, the administration method to a living body is not particularly limited, but it is preferable to use any one or more administration methods of oral administration, vascular administration, subcutaneous administration, rectal administration, application, and drug delivery system.

  Moreover, this invention relates to the health food containing the above-mentioned angiogenesis inhibitor, and a cosmetic composition.

  Carnosic acid is a substance extracted from rosemary and sage, and since it is already sold as a dietary supplement, it has been confirmed to be safe for the human body, especially when administered orally. In addition, the cost is low compared to conventional angiogenesis inhibitors, and long-term administration and ingestion are possible.

  Therefore, according to the present invention, it is possible to provide an angiogenesis inhibitor that overcomes the conventional problems and is extremely safe for the human body and inexpensive.

  Hereinafter, the present invention will be described in detail. However, the scope of the present invention is not limited to these descriptions, and modifications other than the following examples can be made as appropriate without departing from the spirit of the present invention.

  The angiogenesis inhibitor of the present invention contains carnosic acid as an active ingredient, more preferably contains only carnosic acid. Carnosic acid may be extracted from various natural products, chemically synthesized, or commercially available.

  For example, when carnosic acid is extracted from rosemary and / or sage, the part of the target plant to be used is not particularly limited, and any of roots, stems, leaves, petioles, branches, flowers, whole plants, or combinations thereof can be used. It may be used. In the present invention, although not particularly limited, leaves are preferably used. Further, the plant may be either in a raw state or dried by a method well known to those skilled in the art.

  The extract used in the present invention means an extract extracted under appropriate conditions using the above plant as water, a polar or nonpolar solvent, or a mixture thereof.

  The extract of rosemary and / or sage used in the present invention can be obtained, for example, as follows. First, a predetermined part of rosemary and / or sage is immersed in an extraction solvent. The amount of the extraction solvent is not particularly limited as long as the target plant can be immersed therein, but an extraction solvent having a ratio of 2 to 100 times the weight of the target plant is preferable. The type of extraction solvent that can be used is not particularly limited, but examples thereof include lower alcohols such as methanol, ethanol, n-propanol, isopropanol, and t-butanol, ketones such as acetone, and esters such as ethyl acetate. Organic solvents such as ethers, chloroform, and dichloromethane, and water. These may be used alone or in combination. In the present invention, methanol, ethanol, ethyl acetate, or a combination of these solvents and water is preferable. In consideration of administration to a living body, a solvent having low toxicity such as ethanol or a mixed solvent of water and ethanol is more preferable. . Since the immersion time of the target plant can vary depending on various conditions, it is not particularly limited and can be appropriately set by those skilled in the art. In addition, the extraction temperature can vary depending on various conditions, and is not particularly limited, and can be set appropriately by those skilled in the art.

  After the above immersion, the obtained extract may be used as it is, but it may be used in the form of a dried product or a paste obtained by evaporating moisture as necessary. Moreover, in order to refine | purify carnosic acid, you may produce | generate and use as needed using the means (for example, various chromatography) normally used by those skilled in the art.

  Regarding a method for treating a disease caused by angiogenesis using the angiogenesis inhibitor of the present invention, particularly a malignant tumor, the following four steps of angiogenesis process, 1) angiogenesis-promoting factor produced from cancer and stroma Angiogenesis is switched on 2) The basement membrane under vascular endothelial cells is digested by the involvement of MMP and plasminogen activator 3) Vascular endothelial cells migrate and proliferate 4) Vascular endothelial cells enter the lumen Any one or a plurality of them may be suppressed, but the above-described 3) is characterized in that it strongly suppresses the migration and proliferation of vascular endothelial cells. The method of administration to the living body in the above treatment method is not particularly limited because the conditions are changed depending on the treatment method, but any method of oral administration, vascular administration, subcutaneous administration, rectal administration, application, drug delivery system, Alternatively, it is preferable to use a method in which any one or more administration methods are combined.

  Furthermore, the angiogenesis inhibitor of the present invention can take various forms depending on the manner of application. For example, when the angiogenesis inhibitor of the present invention is used as a pharmaceutical composition, its dosage form is not particularly limited, and is processed into an appropriate dosage form according to the method described in the Japanese Pharmacy Law. More specific examples of dosage forms include, for pharmaceutical compositions intended for oral administration, dosage forms such as capsules, tablets, powders, granules, fine granules, sustained release agents, etc. In the case of pharmaceutical compositions intended for parenteral administration, injections intended for intravenous administration, vascular administration such as infusions, or subcutaneous administration such as subcutaneous injection, coating agents such as ointments, suppositories for rectal administration And the like. Furthermore, in administration using a drug delivery system (DDS), nanoparticles such as nanocapsules are also included. The dosage can be easily selected by those skilled in the art because it can be easily changed depending on conditions such as the body weight and dosage form of the subject person. For example, in administration using DDS, the target disease site or cell The concentration of carnosic acid in the region where the group exists is 10 μM to 100 μM, preferably 50 μM to 100 μM.

  In the implementation as the pharmaceutical composition described above, after being processed into the above-mentioned form by a method well known to those skilled in the art, the third component is a diluent such as distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, etc. May be mixed. Furthermore, if necessary, a bactericidal agent, preservative, stabilizer and the like may be mixed.

  As another example, when the angiogenesis inhibitor of the present invention is used as a health food, the form is not limited to a solid food, but includes a beverage (for example, a liquid beverage). More specifically, supplements (forms such as capsules) in the form of liquid, paste, solid, etc., soft drinks, syrups, fresheners in the mouth, jelly, and the like can be mentioned, but the invention is not particularly limited thereto. Such a health food containing the angiogenesis inhibitor of the present invention can be produced using methods well known to those skilled in the art.

  As yet another example, when the angiogenesis inhibitor of the present invention is used as a cosmetic composition, examples of its form include lotions, emulsions, creams, and powders, but are not particularly limited thereto. Such cosmetic compositions can be manufactured using techniques known to those skilled in the art.

  In addition, as described above, the angiogenesis inhibitor of the present invention is produced by a method commonly used in the field according to the embodiment, and an appropriate amount can be taken and applied as appropriate according to the method.

Hereinafter, the present invention will be described more specifically based on examples, but the present invention is not limited thereto.

  Example 1 [Evaluation in an angiogenesis measuring system using rat artery piece] Carnosic acid as a sample used was dissolved in dimethylsulfoxide and used. Rats were Wistar males, and those from 6 to 8 weeks of age were purchased from Nippon Charles River and used. Collagen stock solution (CELLMATRIX type Ia) and reconstitution buffer were purchased from Nitta Gelatin. Culture medium 10 × Eagle's MEM and RPMI 1640 were purchased from Gibco. ITS + was purchased from Becton Dickinson Bioscience.

  A collagen solution was prepared by mixing 8: 1: 1 collagen stock solution, 10 × Eagle's MEM medium, and reconstitution buffer, and stored at 4 ° C.

The effect of carnosic acid on angiogenesis was evaluated by an angiogenesis measurement system in which rat arterial pieces were cultured in collagen gel. Wistar rats were anesthetized with diethyl ether and bleeded by cutting the right femoral artery, and then the thoracic aorta was removed and cut to a length of 1 to 1.5 mm. The arterial piece was transferred to a 6-well culture plate, embedded with a collagen gel solution, and gelled at 37 ° C. After gelation, 2 ml of RPMI 1640 medium containing 1% ITS + was added, and the sample solution was further added to the culture solution. Cultivation was performed in a CO ₂ incubator, and the microvascular network formed from the arterial piece on the seventh day was photographed under an inverted microscope, and the image was taken into a computer, and then the length of the microvessel was measured.

  FIG. 1 shows a photograph of a microvascular network taken under a microscope. In the sample cultured in the absence of carnosic acid (Control), the formation of microvessels was observed, but in the presence of 100 μM carnosic acid, the formation of microvessels was completely suppressed.

  FIG. 2 shows a graph comparing the microvessel length on the seventh day of culture for each carnosic acid concentration. From this result, it was shown that the formation of microvessels was significantly suppressed when the concentration of carnosic acid was 50 μM, and the formation of microvessels was completely suppressed at 100 μM.

Example 2 [Effect of carnosic acid on lumen formation of human umbilical vein-derived vascular endothelial cells (HUVEC) on reconstituted basement membrane]
The sample used was carnosic acid dissolved in dimethyl sulfoxide. Human umbilical vein-derived vascular endothelial cells (HUVEC) were purchased from Kurabo Industries and used after 3 to 7 passages. Reconstituted basement membrane gel (Matrigel ™) was purchased from Becton Dickinson Bioscience.

HUVEC was adjusted to 1 × 10 ⁵ cells / ml in HuMedia-EG2 medium (Kurabo). 50 μl of the reconstituted basement membrane gel was added to the wells of a 96-well culture plate and allowed to gel for 1 hour in a CO ₂ incubator. 100 μl of HUVEC suspension containing each concentration of carnosic acid was added on the reconstituted basement membrane gel, and cultured in a CO ₂ incubator for 12 hours. The state of lumen formation by HUVEC was observed with an optical microscope, and the length of the lumen was measured.

  FIG. 3 shows the results of culturing HUVEC in the form of a reconstituted basement membrane gel for 12 hours. Tube formation was observed in the absence of carnosic acid (Control), but tube formation was strongly suppressed in the presence of 50 μM carnosic acid.

  FIG. 4 is a graph showing the length of the lumen length for each carnosic acid concentration. From this result, it was shown that the lumen formation was significantly suppressed when the concentration of carnosic acid was 50 μM, and the lumen formation was completely suppressed when 100 μM.

Example 3 [Influence of carnosic acid on proliferation of human umbilical vein-derived vascular endothelial cells (HUVEC)]
The sample used was carnosic acid dissolved in dimethyl sulfoxide. Human umbilical vein-derived vascular endothelial cells (HUVEC) were purchased from Kurabo Industries and used after 3 to 7 passages. HUVEC was adjusted to 1.5 × 10 ⁴ cells / ml in HuMedia-EG2 medium (Kurabo). 100 μl of a HUVEC suspension containing each concentration of carnosic acid was added to the wells of a 96-well culture plate and cultured for 72 hours in a CO ₂ incubator. Thereafter, all media were replaced with HuMedia-EG2 media and cultured for 1 hour in a CO ₂ incubator. After adding 10 μl of WST-1 reagent (Dojindo) to each well and culturing for 4 hours in a CO ₂ incubator, the absorbance at 450 nm was measured with a microplate reader to compare the number of living cells.

  FIG. 5 is a graph showing the intensity of absorbance at 450 nm for each carnosic acid concentration. That is, this graph is a result of comparing the ratio of the number of cells grown in the presence of each concentration of carnosic acid, with the number of cells grown in the absence of carnosic acid being 100. From this result, it was shown that the growth of HUVEC was significantly suppressed in the presence of 10 μM or more carnosic acid.

Example 4 [Effect of carnosic acid on migration of human umbilical vein-derived vascular endothelial cells (HUVEC)]
The sample used was carnosic acid dissolved in dimethyl sulfoxide. Human umbilical vein-derived vascular endothelial cells (HUVEC) were purchased from Kurabo Industries and used after 3 to 7 passages. 24-well inserts (8 μm pore size) were purchased from Becton Dickinson Bioscience.

HUVEC was adjusted to 2.5 × 10 ⁵ cells / ml in Medium 199 (Invitrogen) medium (medium A) containing 0.1% bovine serum albumin. 400 μl of medium was added to the wells of a 24-well culture plate. The prepared medium contains medium A, medium A containing 10 ng / ml VEGF and each concentration of carnosic acid. 400 μl of HUVEC suspension was added to a 24-well insert and placed in each well. Incubation was performed in a CO ₂ incubator for 6 hours. The HUVEC remaining on the film on the upper surface of the insert was removed with a cotton swab, the HUVEC moved to the back surface of the film was fixed and stained, and then observed with an optical microscope, and the number of moved HUVECs was counted.

  FIG. 6 is a graph showing the results of counting the number of HUVECs that had moved to the back side of the filter after 6 hours of culture at an arbitrary 5 points under an optical microscope (200 times). From this result, it was shown that migration of HUVEC was significantly suppressed by causing 10 μM or more of carnosic acid to act.

As explained above, the present invention has been confirmed to be safe for the human body, particularly when orally administered. Furthermore, since it has an economically excellent effect that the production cost is lower than that of a conventional angiogenesis inhibitor, it has a feature that enables long-term administration and inoculation. Therefore, according to the present invention, it is possible to provide an angiogenesis inhibitor that is extremely safe for the human body and that is inexpensive and overcomes the conventional problems.

Effect of carnosic acid in angiogenesis measurement system using rat arterial piece. Microvessel growth is seen in the absence of carnosic acid. On the other hand, microvessel formation was completely suppressed in the presence of carnosic acid (100 μM). Day 7 of culture. Effect of carnosic acid in angiogenesis measurement system using rat arterial piece. Results of comparison of microvessel length on day 7 of culture. Values are mean ± standard deviation (n = 6). *: Significantly different from the subject (p <0.01) Effect of carnosic acid on lumen formation of human umbilical vein-derived vascular endothelial cells (HUVEC) on reconstituted basement membrane. Results of seeding HUVEC on reconstituted basement membrane gel and culturing for 12 h. Effect of carnosic acid on lumen formation of human umbilical vein-derived vascular endothelial cells (HUVEC) on reconstituted basement membrane. Results of measuring and comparing lumen lengths. Values are mean ± standard deviation (n = 3-4). *: Significant difference compared to the subject (p <0.01). Effect of carnosic acid on human umbilical vein-derived vascular endothelial cell (HUVEC) proliferation. After culturing 0-100 μM carnosic acid and HUVEC for 72 hours, WST-1 reagent was added and the absorbance at 450 nm was measured. Absorbance in the absence of carnosic acid was calculated as 100 percent. Values are mean ± standard deviation (n = 6). *: Significantly different from the target (p <0.01) Effect of carnosic acid on HUVEC migration. The number of HUVECs that had moved to the back side of the filter after 6 hours of culture was counted at an arbitrary 5 points under an optical microscope (200 times). Values are mean ± standard deviation (n = 3). NC (negative control); VEGF and carnosic acid absence conditions. PC (positive control); Carnosic acid is absent in the presence of VEGF. *: Significantly different from the target (p <0.01)

Claims (8)

  Angiogenesis inhibitor containing carnosic acid as active ingredient   Angiogenesis inhibitor containing only carnosic acid as active ingredient   A method for treating a disease caused by angiogenesis using the angiogenesis inhibitor according to claim 1 or 2.   The method according to claim 3, wherein the disease caused by angiogenesis is a malignant tumor.   The method according to claim 3 or 4, which suppresses migration or proliferation of vascular endothelial cells.   6. The angiogenesis inhibitor is administered using at least one of oral administration, vascular administration, subcutaneous administration, rectal administration, application, and drug delivery system. Method described in   A health food containing the angiogenesis inhibitor according to claim 1 or 2. Cosmetic composition containing the angiogenesis inhibitor according to claim 1 or 2.