JP7526758B2 - Composition comprising safflower extract for stabilizing blood vessels in a subject - Google Patents
Composition comprising safflower extract for stabilizing blood vessels in a subject Download PDFInfo
- Publication number
- JP7526758B2 JP7526758B2 JP2022089908A JP2022089908A JP7526758B2 JP 7526758 B2 JP7526758 B2 JP 7526758B2 JP 2022089908 A JP2022089908 A JP 2022089908A JP 2022089908 A JP2022089908 A JP 2022089908A JP 7526758 B2 JP7526758 B2 JP 7526758B2
- Authority
- JP
- Japan
- Prior art keywords
- composition
- safflower extract
- endothelial cells
- blood vessels
- subject
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000203 mixture Substances 0.000 title claims description 77
- 229940119485 safflower extract Drugs 0.000 title claims description 43
- 210000004204 blood vessel Anatomy 0.000 title claims description 34
- 230000000087 stabilizing effect Effects 0.000 title claims description 16
- 210000003556 vascular endothelial cell Anatomy 0.000 claims description 36
- 108010018828 cadherin 5 Proteins 0.000 claims description 24
- 230000014509 gene expression Effects 0.000 claims description 21
- 102000008790 VE-cadherin Human genes 0.000 claims description 20
- 239000000243 solution Substances 0.000 claims description 15
- 230000000699 topical effect Effects 0.000 claims description 13
- 239000006185 dispersion Substances 0.000 claims description 10
- 239000000839 emulsion Substances 0.000 claims description 10
- 239000006072 paste Substances 0.000 claims description 10
- 239000000725 suspension Substances 0.000 claims description 10
- -1 blocks Substances 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 239000002537 cosmetic Substances 0.000 claims description 6
- 239000002775 capsule Substances 0.000 claims description 5
- 239000006071 cream Substances 0.000 claims description 5
- 239000006260 foam Substances 0.000 claims description 5
- 239000000499 gel Substances 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 5
- 239000006210 lotion Substances 0.000 claims description 5
- 239000002674 ointment Substances 0.000 claims description 5
- 239000006187 pill Substances 0.000 claims description 5
- 239000007921 spray Substances 0.000 claims description 5
- 239000003826 tablet Substances 0.000 claims description 5
- 230000017531 blood circulation Effects 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 20
- 239000002609 medium Substances 0.000 description 20
- 230000002792 vascular Effects 0.000 description 19
- 230000000694 effects Effects 0.000 description 14
- 101100481408 Danio rerio tie2 gene Proteins 0.000 description 12
- 101100481410 Mus musculus Tek gene Proteins 0.000 description 12
- 238000000034 method Methods 0.000 description 12
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 10
- 229910001424 calcium ion Inorganic materials 0.000 description 10
- 210000002889 endothelial cell Anatomy 0.000 description 7
- 244000020518 Carthamus tinctorius Species 0.000 description 6
- 235000003255 Carthamus tinctorius Nutrition 0.000 description 6
- 239000003102 growth factor Substances 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 5
- 230000004913 activation Effects 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 229910052791 calcium Inorganic materials 0.000 description 5
- 239000011575 calcium Substances 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 230000006453 vascular barrier function Effects 0.000 description 4
- 230000008728 vascular permeability Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 102000008186 Collagen Human genes 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 229920001436 collagen Polymers 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 210000001365 lymphatic vessel Anatomy 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 210000003491 skin Anatomy 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- 229940058015 1,3-butylene glycol Drugs 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 2
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 235000019437 butane-1,3-diol Nutrition 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- 230000035800 maturation Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 230000002093 peripheral effect Effects 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 108091008598 receptor tyrosine kinases Proteins 0.000 description 2
- 102000027426 receptor tyrosine kinases Human genes 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 238000012353 t test Methods 0.000 description 2
- 210000003606 umbilical vein Anatomy 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 102000009088 Angiopoietin-1 Human genes 0.000 description 1
- 108010048154 Angiopoietin-1 Proteins 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 241000208838 Asteraceae Species 0.000 description 1
- 229920001342 Bakelite® Polymers 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 238000011530 RNeasy Mini Kit Methods 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 102000012753 TIE-2 Receptor Human genes 0.000 description 1
- 108010090091 TIE-2 Receptor Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000003974 emollient agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 210000001650 focal adhesion Anatomy 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- VPSRLGDRGCKUTK-UHFFFAOYSA-N fura-2-acetoxymethyl ester Chemical compound CC(=O)OCOC(=O)CN(CC(=O)OCOC(C)=O)C1=CC=C(C)C=C1OCCOC(C(=C1)N(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=CC2=C1OC(C=1OC(=CN=1)C(=O)OCOC(C)=O)=C2 VPSRLGDRGCKUTK-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000008123 high-intensity sweetener Substances 0.000 description 1
- 230000000887 hydrating effect Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 210000005073 lymphatic endothelial cell Anatomy 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 235000013615 non-nutritive sweetener Nutrition 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 235000018343 nutrient deficiency Nutrition 0.000 description 1
- 239000003605 opacifier Substances 0.000 description 1
- 210000003668 pericyte Anatomy 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000000865 phosphorylative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000008326 skin blood flow Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
- 230000006438 vascular health Effects 0.000 description 1
- 210000004509 vascular smooth muscle cell Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/286—Carthamus (distaff thistle)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Botany (AREA)
- Heart & Thoracic Surgery (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Vascular Medicine (AREA)
- Cardiology (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Description
本発明は、被験体において血管を安定化させるための、ベニバナエキスを含んでなる組成物に関する。 The present invention relates to a composition comprising safflower extract for stabilizing blood vessels in a subject.
生命がその活動を正常に維持するためには、各臓器へ十分な酸素や栄養を供給し、さらにその活動で生じる老廃物を適切に運搬・処理することが重要であるが、この機能を担う器官が血管である。血管は、血液を通して全身の臓器に酸素や栄養素を送るだけでなく、組織中からの老廃物を運び体外へ排泄する役割を担っており、人体が生命活動を維持するにあたって重要な組織である。 For life to maintain normal activity, it is important to supply each organ with sufficient oxygen and nutrients, and to properly transport and process waste products generated by these activities; the organ that performs this function is the blood vessels. Blood vessels not only deliver oxygen and nutrients to all organs throughout the body through blood, but also transport waste products from tissues and excrete them from the body, making them an important tissue for the human body to maintain vital functions.
血管には、心臓から送り出された血液が流れる動脈、心臓へ戻る血液が通る静脈の他に、血管のうち９０％以上（人間の場合）を占める毛細血管がある。毛細血管は主に血管内皮細胞と血管壁細胞により構成される。 In addition to arteries, which carry blood sent from the heart, and veins, which carry blood back to the heart, there are also capillaries, which account for more than 90% of blood vessels (in humans). Capillaries are mainly composed of vascular endothelial cells and vascular wall cells.
血管内皮細胞は、例えば、互いに接着し血管内環境因子（細胞および液性因子）が容易には血管外に漏出しないような内皮細胞間の接着斑を形成すること等によって、血管のバリアとして働き、また、物質の透過性すなわちバリア機能を調節することで、生命の健康維持に重要な役割を担うことが近年分かってきた。血管バリア機能の障害により、炎症、動脈硬化など様々な血管の病気の引き金となるだけではなく、末梢細胞への酸素、栄養の不足が生じることが知られている。 It has been discovered in recent years that vascular endothelial cells act as a vascular barrier by, for example, adhering to each other and forming focal adhesions between endothelial cells that prevent intravascular environmental factors (cellular and humoral factors) from easily leaking out of the blood vessels, and also play an important role in maintaining vital health by regulating the permeability of substances, i.e., the barrier function. It is known that impaired vascular barrier function not only triggers various vascular diseases such as inflammation and arteriosclerosis, but also causes oxygen and nutrient deficiencies in peripheral cells.
また、血管壁細胞と総称される血管平滑筋細胞やペリサイトは、血管内皮細胞の外腔面から、細胞外マトリックスを介してまたは直接的に血管内皮細胞に接着している。血管径の大きさによっては血管内皮細胞と血管壁細胞の接着する比率は異なり、大血管系では、１層の血管内皮細胞に、複数層の血管壁細胞が裏打ちしており、中、小血管では、血管内皮細胞１細胞に血管壁細胞が１細胞裏打ちするものが多く、また、さらに径の細い血管では、１個の血管壁細胞が複数の血管内皮細胞に接着している。このように、血管壁細胞が血管内皮細胞に対し裏打ちすることは血管の構造上重要である。例えば、組織の酸素や栄養分の需要に応じて、とくにそれらが不足する際には、血管は管腔を拡大化させることで血流を増加させ、酸素、養分を組織に十分に行き渡らせるように調節することが可能である。 Vascular smooth muscle cells and pericytes, collectively known as vascular wall cells, adhere to vascular endothelial cells from the outer surface of the vascular endothelial cells, either directly or via the extracellular matrix. The ratio of adhesion between vascular endothelial cells and vascular wall cells varies depending on the size of the blood vessel diameter. In large vascular systems, multiple layers of vascular wall cells line one layer of vascular endothelial cells, while in medium and small blood vessels, one vascular endothelial cell is often lined by one vascular wall cell, and in blood vessels with even smaller diameters, one vascular wall cell adheres to multiple vascular endothelial cells. In this way, vascular wall cells lining vascular endothelial cells is important for the structure of blood vessels. For example, depending on the tissue's demand for oxygen and nutrients, especially when these are insufficient, blood vessels can increase blood flow by expanding the lumen, and adjust so that oxygen and nutrients are sufficiently distributed to the tissues.
この血管のバリア機能に影響を与える因子としては、例えば、血管内皮細胞に発現する受容体型チロシンキナーゼであるＴｉｅ２（Ｔｙｒｏｓｉｎｅ ｋｉｎａｓｅ ｗｉｔｈ Ｉｇ ａｎｄ ＥＧＦ ｈｏｍｏｌｏｇｙ ｄｏｍａｉｎ２）があり、Ｔｉｅ２を活性化することにより、血管が成熟化および安定化すること等が知られている。 One example of a factor that affects the barrier function of blood vessels is Tie2 (tyrosine kinase with Ig and EGF homology domain 2), a receptor tyrosine kinase expressed in vascular endothelial cells. It is known that activating Tie2 leads to the maturation and stabilization of blood vessels.
より具体的には、例えば、非特許文献１において、Ａｎｇ－１が、Ｔｉｅ２受容体を介して、内皮細胞間および内皮細胞－壁細胞間接着を増強し、血管安定化と血管透過性の低下を引き起こすこと、Ａｎｇ－１がｉｎ ｖｉｔｒｏの内皮細胞において、ＶＥカドヘリン接着を増強し、血管透過性を低下させること、およびＡｎｇ－１－Ｔｉｅ２シグナルはＶＥカドヘリン接着を増強し、血管バリア機能を制御していると考えられることが記載されている。 More specifically, for example, Non-Patent Document 1 describes that Ang-1 enhances adhesion between endothelial cells and between endothelial cells and mural cells via the Tie2 receptor, resulting in vascular stabilization and reduced vascular permeability, that Ang-1 enhances VE-cadherin adhesion and reduces vascular permeability in endothelial cells in vitro, and that the Ang-1-Tie2 signal enhances VE-cadherin adhesion and is thought to control vascular barrier function.
本発明者らは、ベニバナエキスが、被験体において血管を安定化させることを見出した。本発明はこの知見に基づくものである。 The present inventors have found that safflower extract stabilizes blood vessels in subjects. The present invention is based on this finding.
よって、本発明は、被験体において血管を安定化させるための、ベニバナエキスを含んでなる組成物を提供する。 The present invention therefore provides a composition comprising safflower extract for stabilizing blood vessels in a subject.
本発明によれば、以下の発明が提供される。
（１）被験体において血管を安定化させるための組成物であって、ベニバナエキスを含んでなる、組成物。
（２）血管内皮細胞においてＶＥカドヘリンの発現を上昇させるための、（１）に記載の組成物。
（３）血管内皮細胞内のカルシウムイオン濃度を上昇させるための、（１）または（２）に記載の組成物。
（４）前記被験体がヒト被験体である、（１）～（３）のいずれかに記載の組成物。
（５）経口用または局所用の組成物である、（１）～（４）のいずれかに記載の組成物。
（６）経口用の組成物が、溶液、懸濁液、分散液、エマルジョン、ペースト、粉末、顆粒、丸剤、錠剤、ブロック、またはカプセルの形態である、（５）に記載の組成物。
（７）局所用の組成物が、溶液、懸濁液、分散液、エマルジョン、クリーム、軟膏、ゲル、ローション、フォーム、ペースト、またはスプレーの形態である、（５）に記載の組成物。
（８）化粧料組成物である、（１）～（７）のいずれかに記載の組成物。
（９）被験体における血管の安定化を目的とする薬剤の製造のための、ベニバナエキスの使用。
（１０）血管内皮細胞におけるＶＥカドヘリン発現の上昇を目的とする薬剤の製造のための、（９）の使用。
（１１）血管内皮細胞内のカルシウムイオン濃度の上昇を目的とする薬剤の製造のための、（９）または（１０）に記載の使用。
（１２）前記被験体がヒト被験体である、（９）～（１１）のいずれかに記載の使用。
（１３）ベニバナエキスを含んでなる組成物の、被験体において血管を安定化させるための使用。
（１４）血管内皮細胞においてＶＥカドヘリンの発現を上昇させるための、（１３）に記載の使用。
（１５）血管内皮細胞内のカルシウムイオン濃度を上昇させるための、（１３）または（１４）に記載の使用。
（１６）前記被験体がヒト被験体である、（１３）～（１５）のいずれかに記載の使用。
（１７）前記組成物が経口用または局所用の組成物である、（１３）～（１６）のいずれかに記載の使用。
（１８）前記経口用の組成物が、溶液、懸濁液、分散液、エマルジョン、ペースト、粉末、顆粒、丸剤、錠剤、ブロック、またはカプセルの形態である、（１７）に記載の使用。
（１９）前記局所用の組成物が、溶液、懸濁液、分散液、エマルジョン、クリーム、軟膏、ゲル、ローション、フォーム、ペースト、またはスプレーの形態である、（１７）に記載の使用。
（２０）前記組成物が化粧料組成物である、（１３）～（１９）のいずれかに記載の使用。
（２１）被験体において、血管を安定化させるための方法であって、ベニバナエキスを含んでなる組成物の有効量を投与することを含む、方法。
（２２）血管内皮細胞においてＶＥカドヘリンの発現を上昇させるための、（２１）に記載の方法。
（２３）血管内皮細胞内のカルシウムイオン濃度を上昇させるための、（２１）または（２２）に記載の方法。
（２４）前記被験体がヒト被験体である、（２１）～（２３）のいずれかに記載の方法。
（２５）前記組成物が経口用または局所用の組成物である、（２１）～（２４）のいずれかに記載の方法。
（２６）前記経口用の組成物が、溶液、懸濁液、分散液、エマルジョン、ペースト、粉末、顆粒、丸剤、錠剤、ブロック、またはカプセルの形態である、（２５）に記載の方法。
（２７）前記局所用の組成物が、溶液、懸濁液、分散液、エマルジョン、クリーム、軟膏、ゲル、ローション、フォーム、ペースト、またはスプレーの形態である、（２５）に記載の方法。
（２８）前記組成物が化粧料組成物である、（２１）～（２７）のいずれかに記載の方法。
According to the present invention, the following inventions are provided.
(1) A composition for stabilizing blood vessels in a subject, the composition comprising safflower extract.
(2) The composition described in (1) for increasing the expression of VE-cadherin in vascular endothelial cells.
(3) The composition according to (1) or (2) for increasing calcium ion concentration in vascular endothelial cells.
(4) The composition according to any one of (1) to (3), wherein the subject is a human subject.
(5) The composition according to any one of (1) to (4), which is an oral or topical composition.
(6) The composition according to (5), wherein the oral composition is in the form of a solution, suspension, dispersion, emulsion, paste, powder, granules, pills, tablets, blocks, or capsules.
(7) The composition according to (5), wherein the topical composition is in the form of a solution, suspension, dispersion, emulsion, cream, ointment, gel, lotion, foam, paste, or spray.
(8) The composition according to any one of (1) to (7), which is a cosmetic composition.
(9) Use of safflower extract for the manufacture of a medicament for stabilizing blood vessels in a subject.
(10) Use of (9) for the manufacture of a drug for increasing VE-cadherin expression in vascular endothelial cells.
(11) The use according to (9) or (10) for the manufacture of a drug intended to increase calcium ion concentration in vascular endothelial cells.
(12) The use according to any one of (9) to (11), wherein the subject is a human subject.
(13) Use of a composition comprising safflower extract for stabilizing blood vessels in a subject.
(14) The use according to (13) for increasing expression of VE-cadherin in vascular endothelial cells.
(15) The use according to (13) or (14) for increasing calcium ion concentration in vascular endothelial cells.
(16) The use according to any one of (13) to (15), wherein the subject is a human subject.
(17) The use according to any one of (13) to (16), wherein the composition is an oral or topical composition.
(18) The use according to (17), wherein the oral composition is in the form of a solution, suspension, dispersion, emulsion, paste, powder, granules, pills, tablets, blocks, or capsules.
(19) The use according to (17), wherein the topical composition is in the form of a solution, suspension, dispersion, emulsion, cream, ointment, gel, lotion, foam, paste, or spray.
(20) The use according to any one of (13) to (19), wherein the composition is a cosmetic composition.
(21) A method for stabilizing blood vessels in a subject, comprising administering an effective amount of a composition comprising safflower extract.
(22) The method according to (21) for increasing expression of VE-cadherin in vascular endothelial cells.
(23) The method according to (21) or (22) for increasing calcium ion concentration in vascular endothelial cells.
(24) The method according to any one of (21) to (23), wherein the subject is a human subject.
(25) The method according to any one of (21) to (24), wherein the composition is an oral or topical composition.
(26) The method according to (25), wherein the oral composition is in the form of a solution, suspension, dispersion, emulsion, paste, powder, granules, pills, tablets, blocks, or capsules.
(27) The method according to (25), wherein the topical composition is in the form of a solution, suspension, dispersion, emulsion, cream, ointment, gel, lotion, foam, paste, or spray.
(28) The method according to any one of (21) to (27), wherein the composition is a cosmetic composition.
本発明によれば、被験体において血管を安定化させるための、ベニバナエキスを含んでなる組成物を提供することができる。この組成物は、血管内皮細胞においてＶＥカドヘリンの発現を上昇させることも可能である。また、この組成物は、血管内皮細胞内のカルシウムイオン濃度を上昇させることも可能である。 According to the present invention, a composition comprising safflower extract for stabilizing blood vessels in a subject can be provided. This composition can also increase the expression of VE-cadherin in vascular endothelial cells. This composition can also increase the calcium ion concentration in vascular endothelial cells.
本発明によれば、ベニバナエキスを含んでなる組成物が提供され、該組成物は、被験体において血管を安定化させることが可能である。 According to the present invention, a composition comprising safflower extract is provided, which is capable of stabilizing blood vessels in a subject.
本発明の好ましい実施態様によれば、本発明の組成物は、血管内皮細胞においてＶＥカドヘリンの発現を上昇させるためのものとされる。本発明の別の好ましい実施態様によれば、本発明の組成物は、血管内皮細胞内のカルシウムイオン濃度を上昇させるためのものとされる。 According to a preferred embodiment of the present invention, the composition of the present invention is intended to increase the expression of VE-cadherin in vascular endothelial cells. According to another preferred embodiment of the present invention, the composition of the present invention is intended to increase the calcium ion concentration in vascular endothelial cells.
本発明における「ベニバナ」は、キク科植物のベニバナ（Ｃａｒｔｈａｍｕｓ ｔｉｎｃｔｏｒｉｕｓ Ｌ．）を意味する。使用部位としては、限定されるわけではないが、葉、茎、幹、花、根、果実等が挙げられ、好ましくは花である。 In the present invention, "safflower" refers to safflower (Carthamus tinctorius L.), a plant of the Asteraceae family. Parts to be used include, but are not limited to, leaves, stems, trunks, flowers, roots, fruits, etc., with flowers being preferred.
本発明における「ベニバナ」の産地としては、限定されるわけではないが、中国、韓国、日本国、イスラエル、アメリカ、インド等が挙げられる。また、本発明における「ベニバナ」の仕入れ時期も限定されず、いかなる仕入れ時期のものを使用してもよい。 The origin of the "safflower" in the present invention includes, but is not limited to, China, Korea, Japan, Israel, the United States, India, etc. Furthermore, the purchase time of the "safflower" in the present invention is not limited, and safflower purchased at any time may be used.
本発明におけるベニバナエキスの製造方法は、特に制限されず、該技術分野において通常使用される方法に応じて抽出することができる。前記抽出方法としては、限定されるわけではないが、例えば、熱水抽出法、超音波抽出法、ろ過法、還流抽出法、溶媒抽出法等が挙げられる。これらは単独で実行、または２種以上の方法を併用して行ってもよい。また、高純度のエキスを得るためにエキスを同様の方法で１回以上ずつさらに抽出してもよい。 The method for producing the safflower extract in the present invention is not particularly limited, and extraction can be performed according to a method commonly used in the technical field. The extraction method is not limited to, but may be, for example, hot water extraction, ultrasonic extraction, filtration, reflux extraction, solvent extraction, etc. These may be performed alone, or two or more methods may be used in combination. In addition, the extract may be further extracted one or more times in the same manner to obtain a high-purity extract.
本発明におけるベニバナエキスの製造のために使用される抽出溶媒の種類は特に制限されず、本発明の組成物に本発明の効果を付するベニバナエキスが得られるものであれば、該技術分野において公知となっている任意の溶媒を使用してもよい。そのような溶媒の例としては、限定されるわけではないが、例えば、水、メタノール、エタノール、イソプロパノール、アセトン、１，３－ブチレングリコール、エチレングリコール、プロピレングリコール、グリセリン、酢酸、酢酸エチル、エーテル、ヘキサン等が挙げられ、これらは２つ以上組み合わせて使用してもよい。 The type of extraction solvent used to produce the safflower extract of the present invention is not particularly limited, and any solvent known in the technical field may be used as long as it produces a safflower extract that imparts the effects of the present invention to the composition of the present invention. Examples of such solvents include, but are not limited to, water, methanol, ethanol, isopropanol, acetone, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, acetic acid, ethyl acetate, ether, hexane, etc., and two or more of these may be used in combination.
ベニバナエキスが商業的に販売されている場合、本発明の組成物に本発明の効果を付するものであれば、それを使用してもよい。 If safflower extract is commercially available, it may be used if it imparts the effects of the present invention to the composition of the present invention.
本発明の組成物に含まれるベニバナエキスの配合量は、用途に応じて適宜決定できる。 The amount of safflower extract contained in the composition of the present invention can be determined appropriately depending on the application.
本発明の組成物は、限定されるわけではないが、化粧料組成物としてもよい。本発明における化粧料組成物とは、化粧をすることを目的に使用される組成物のことを意味し、経口用組成物、局所用組成物のいずれであってもよい。また、本発明の組成物としては、限定されるわけではないが、医薬組成物、食品組成物等も挙げられる。 The composition of the present invention may be, but is not limited to, a cosmetic composition. In the present invention, a cosmetic composition means a composition used for the purpose of applying makeup, and may be either an oral composition or a topical composition. In addition, the composition of the present invention may be, but is not limited to, a pharmaceutical composition, a food composition, etc.
本発明の組成物は、経口用組成物、局所用組成物のいずれであってもよい。 The composition of the present invention may be either an oral composition or a topical composition.
本発明の組成物を経口用組成物とする場合、経口用組成物に配合される公知の成分（添加剤）をさらに配合することができる。このような成分としては、限定されるわけではないが、例えば、甘味料、高糖味度甘味料、香料、着色料、酸味料、抗酸化剤、乳化剤、防腐剤、安定剤、溶媒、賦形剤、結合剤、滑沢剤、充填剤、粉末、ビタミン、アミノ酸等が挙げられる。 When the composition of the present invention is used as an oral composition, it may further contain known components (additives) that are incorporated into oral compositions. Examples of such components include, but are not limited to, sweeteners, high-intensity sweeteners, flavors, colorants, acidulants, antioxidants, emulsifiers, preservatives, stabilizers, solvents, excipients, binders, lubricants, fillers, powders, vitamins, amino acids, etc.
本発明における経口用組成物は、液体、半固体、固体のいずれであってもよく、限定されるわけではないが、好ましくは、溶液、懸濁液、分散液、エマルジョン、ペースト、粉末、顆粒、丸剤、錠剤、ブロック、またはカプセルの形態である。 The oral composition of the present invention may be liquid, semi-solid, or solid, and is preferably in the form of, but not limited to, a solution, suspension, dispersion, emulsion, paste, powder, granules, pills, tablets, blocks, or capsules.
本発明の組成物を局所用組成物とする場合、局所用組成物に配合される公知の成分（添加剤）をさらに配合することができる。このような成分としては、限定されるわけではないが、例えば、乳化剤、水和剤、溶媒、エモリエント、安定剤、増粘剤、保存剤、滑沢剤、キレート剤、充填剤、賦形剤、粉末、芳香剤、香料、吸収剤、染料、乳白剤、抗酸化剤、ビタミン、アミノ酸等が挙げられる。 When the composition of the present invention is used as a topical composition, it may further contain known ingredients (additives) that are used in topical compositions. Examples of such ingredients include, but are not limited to, emulsifiers, hydrating agents, solvents, emollients, stabilizers, thickeners, preservatives, lubricants, chelating agents, fillers, excipients, powders, fragrances, flavorings, absorbents, dyes, opacifiers, antioxidants, vitamins, amino acids, etc.
本発明における局所用組成物は、液体、半固体、固体のいずれであってもよく、限定されるわけではないが、好ましくは、溶液、懸濁液、分散液、エマルジョン、クリーム、軟膏、ゲル、ローション、フォーム、ペースト、またはスプレーの形態である。 The topical compositions of the present invention may be liquid, semi-solid, or solid, and are preferably in the form of, but not limited to, a solution, suspension, dispersion, emulsion, cream, ointment, gel, lotion, foam, paste, or spray.
本発明における被験体は、いかなる動物であってもよく、限定されるわけではないが、好ましくは哺乳動物、例えば、ヒト、チンパンジーなどの霊長類、イヌ、ネコなどのペット動物、ウシ、ウマ、ヒツジ、ヤギなどの家畜動物、マウス、ラットなどの齧歯類等の哺乳動物とされ、より好ましくはヒトとされる。 The subject in the present invention may be any animal, and is not limited to any particular animal, but is preferably a mammal, for example, a primate such as a human or chimpanzee, a pet animal such as a dog or cat, a livestock animal such as a cow, horse, sheep or goat, or a rodent such as a mouse or rat, and is more preferably a human.
本発明の組成物の「血管の安定化」という用途は、直接的にまたは間接的に、血管を安定な状態にすることを意味し、血管の状態が健常者のそれと同等でない対象の血管の状態が、健常者と同等の血管の状態に戻るもしくは近づくこと、または血管の状態が健常者のそれと同等である対象の血管の状態が、維持もしくは健常者と同等の血管の状態よりも良好になることをいう。該血管が存在する場所は皮膚をはじめ身体のどこであってよい。また、本発明の組成物の「血管の安定化」という用途は、血管における細胞同士の接着を調節することの有無によらず達成され、限定されるわけではないが、例えば、Ｔｉｅ２活性化、血管透過性抑制、血管の成熟化、血管の正常化、血管の健全化、血管のバリア機能の強化等の作用によって達成されてもよい The use of the composition of the present invention to "stabilize blood vessels" means to directly or indirectly stabilize blood vessels, and refers to the return or approach of the vascular state of a subject whose vascular state is not equivalent to that of a healthy person to the vascular state of a healthy person, or the maintenance or improvement of the vascular state of a subject whose vascular state is equivalent to that of a healthy person. The blood vessels may be present anywhere in the body, including the skin. In addition, the use of the composition of the present invention to "stabilize blood vessels" can be achieved regardless of whether or not it regulates adhesion between cells in blood vessels, and is not limited to this use, and may be achieved, for example, by the action of Tie2 activation, vascular permeability inhibition, vascular maturation, vascular normalization, vascular health, and vascular barrier function enhancement.
一般的に、「Ｔｉｅ２」とは、受容体型チロシンキナーゼ（Ｔｙｒｏｓｉｎｅ ｋｉｎａｓｅ ｗｉｔｈ Ｉｇ ａｎｄ ＥＣＦ ｈｏｍｏｌｏｇｙ ｄｏｍａｉｎ ２）を意味する。本発明において「Ｔｉｅ２活性化」とは、Ｔｉｅ２をリン酸化することで、その活性体（リン酸化Ｔｉｅ２）に変換できる能力をいう。毛細血管の構造の安定化には血管内皮細胞に存在するＴｉｅ２と呼ばれる受容体の活性化が重要な役割を果たしている。このＴｉｅ２は血管内皮細胞のほかリンパ管内皮細胞にも存在が示され、血管壁細胞等から放出されるアンジオポエチン－１によって活性化され、血管内皮細胞同士の接着や血管壁細胞との接着を強め、血管構造の安定化に関与している。このＴｉｅ２の活性は、例えば、年齢と共に低下するという知見も報告され、それに伴い血管やリンパ管も老化することが明らかとなっている。また、Ｔｉｅ２活性化により血管内皮細胞の安定化、血管透過性の抑制等が観察される。よって、例えば、血管内皮細胞が強固に結びつくことにより肌に栄養が行き渡り、肌のハリやキメの改善、しわ防止等をすることも可能である。また、血管やリンパ管の内皮細胞の隙間が塞がれることにより血流が改善し、組織中からの水分や老廃物が回収されて、冷え、肩こり、むくみ、クマなどの諸症状を改善、予防することも可能である。したがって、Ｔｉｅ２活性化作用により血管やリンパ管の状態を改善することによって、本発明の「血管の安定化」という用途を達成し得る。 In general, "Tie2" means a receptor tyrosine kinase (tyrosine kinase with Ig and ECF homology domain 2). In the present invention, "Tie2 activation" refers to the ability to convert Tie2 into its active form (phosphorylated Tie2) by phosphorylating it. Activation of a receptor called Tie2 present in vascular endothelial cells plays an important role in stabilizing the structure of capillaries. This Tie2 has been shown to be present not only in vascular endothelial cells but also in lymphatic endothelial cells, and is activated by angiopoietin-1 released from vascular wall cells, etc., strengthening adhesion between vascular endothelial cells and between vascular wall cells, and is involved in stabilizing the vascular structure. It has also been reported that the activity of this Tie2 decreases with age, and it has become clear that blood vessels and lymphatic vessels also age accordingly. Furthermore, Tie2 activation is observed to stabilize vascular endothelial cells, inhibit vascular permeability, and the like. Therefore, for example, by firmly binding vascular endothelial cells, nutrients are distributed to the skin, which can improve the firmness and texture of the skin and prevent wrinkles. Furthermore, by blocking the gaps between endothelial cells in blood vessels and lymphatic vessels, blood flow is improved and moisture and waste products are collected from the tissue, making it possible to improve and prevent various symptoms such as chills, stiff shoulders, swelling, and dark circles under the eyes. Therefore, by improving the condition of blood vessels and lymphatic vessels through Tie2 activation, the purpose of the present invention, "stabilizing blood vessels," can be achieved.
また、血管の安定化によって血流が改善されることから、本発明の組成物を適用した被験体では血流改善が生じる。したがって、本発明の組成物は、血流改善のためにも使用することができる。 In addition, since blood flow is improved by stabilizing blood vessels, improvement in blood flow occurs in subjects to whom the composition of the present invention is applied. Therefore, the composition of the present invention can also be used to improve blood flow.
本発明において血流改善とは、血液の流れが改善されることをいい、血流レベルが健常者のそれと同等でない対象の血流レベルが、健常者と同等の血流レベルに戻るまたは近づくことをいう。これは、正常時の血流レベルと同等でない血流レベルにある対象の血流レベルが、正常時の血流レベルに戻ることであってもよい。被験体に本発明の組成物を適用してから、健常者と同等の血流レベルまたは正常時の血流レベルに戻るまでの時間は短いことが好ましい。血流は血行と称することもある。血流は、限定されるわけではないが、例えば、血流計（例えば、超音波血流計、レーザー血流計など）、位相コントラストＭＲＩ、Ｖｅｃｔｏｒ Ｆｌｏｗ Ｍａｐｐｉｎｇ、Ｅｃｈｏ ＰＩＶなどを用いて測定することができる。血流は、末梢（例えば、指先）の血流であってよく、皮膚血流であってよい。 In the present invention, improvement of blood flow refers to an improvement in blood flow, and refers to the blood flow level of a subject whose blood flow level is not equivalent to that of a healthy person returning to or approaching the blood flow level of a healthy person. This may mean that the blood flow level of a subject whose blood flow level is not equivalent to the normal blood flow level returns to the normal blood flow level. It is preferable that the time from application of the composition of the present invention to the subject until the blood flow level returns to the same level as a healthy person or the normal blood flow level is short. Blood flow is sometimes referred to as blood circulation. Blood flow can be measured using, for example, a blood flow meter (e.g., an ultrasonic blood flow meter, a laser blood flow meter, etc.), phase contrast MRI, Vector Flow Mapping, Echo PIV, etc., but is not limited thereto. Blood flow may be peripheral (e.g., fingertip) blood flow or skin blood flow.
本発明の組成物の「血管内皮細胞においてＶＥカドヘリンの発現を上昇させる」という用途は、直接的にまたは間接的に、血管内皮細胞におけるＶＥカドヘリンの発現レベルを上昇させることを意味し、本発明においてＶＥカドヘリンは遺伝子（例えば、ｍＲＮＡ、ＤＮＡ等）またはタンパク質のいずれであってもよい。本発明の組成物の「血管内皮細胞においてＶＥカドヘリンの発現を上昇させる」という用途により、「血管内皮細胞内のカルシウムイオン濃度を上昇させる」または「血管の安定化」が達成されてもよいが、これに限定されない。 The use of the composition of the present invention to "increase the expression of VE-cadherin in vascular endothelial cells" means directly or indirectly increasing the expression level of VE-cadherin in vascular endothelial cells, and in the present invention, VE-cadherin may be either a gene (e.g., mRNA, DNA, etc.) or a protein. The use of the composition of the present invention to "increase the expression of VE-cadherin in vascular endothelial cells" may achieve, but is not limited to, "increasing the calcium ion concentration in vascular endothelial cells" or "stabilizing blood vessels."
本発明の組成物の「血管内皮細胞内のカルシウムイオン濃度を上昇させる」という用途は、直接的にまたは間接的に、血管内皮細胞内のカルシウムイオンの濃度を上昇させることを意味する。本発明の組成物の「血管内皮細胞内のカルシウムイオン濃度を上昇させる」という用途により、「血管内皮細胞におけるＶＥカドヘリンの発現を上昇させる」または「血管の安定化」が達成されてもよいが、これに限定されない。 The use of the composition of the present invention to "increase calcium ion concentration in vascular endothelial cells" means directly or indirectly increasing the concentration of calcium ions in vascular endothelial cells. The use of the composition of the present invention to "increase calcium ion concentration in vascular endothelial cells" may achieve, but is not limited to, "increasing the expression of VE-cadherin in vascular endothelial cells" or "stabilizing blood vessels."
以下の例に基づいて本発明を具体的に説明するが、本発明はこれらの例に限定されるものではない。含有量は特記しない限り、質量％で示す。 The present invention will be specifically described based on the following examples, but the present invention is not limited to these examples. Contents are expressed in mass % unless otherwise specified.
ヒト臍帯静脈内皮細胞の培養
入手したヒト臍帯静脈内皮細胞（ＨｕＶＥＣ）（ＬＯＮＺＡ社製）を、成長因子（ＬＯＮＺＡ社製）を含むＥＢＭ－２培地（ＬＯＮＺＡ社製）で培養した。ＨｕＶＥＣは、コンフルエント状態の８０～９０％で、４日ごとに継代した。
Cultivation of human umbilical vein endothelial cells Human umbilical vein endothelial cells (HuVEC) (manufactured by LONZA) were cultured in EBM-2 medium (manufactured by LONZA) containing growth factors (manufactured by LONZA). HuVEC were passaged every 4 days at 80-90% confluence.
継代培養は以下の方法により行った。まず、ＨｕＶＥＣを培養しているディッシュからＥＢＭ－２培地を除去し、ディッシュ上のＨｕＶＥＣをＰＢＳで１回洗浄し、洗浄後ＰＢＳを除去した。次に、ディッシュ上のＨｕＶＥＣに１ｍＬの０．０５％のトリプシンＥＤＴＡ（１ｘ）（ｇｉｂｃｏ社製）を添加し、３７℃で３分間インキュベートした。インキュベート後、ディッシュを軽くたたき、ディッシュの底からＨｕＶＥＣを静かに剥がした。成長因子を含有した１０ｍＬのＥＢＭ－２培地をディッシュ添加した後、ディッシュ上のＨｕＶＥＣを１５ｍＬチューブに回収した。１５ｍＬチューブを１０００ｒｐｍ、３分間、室温で遠心分離し、上清を除去した後、１ｍＬの新たなＥＢＭ－２培地を添加してＨｕＶＥＣを懸濁させた。このＨｕＶＥＣを懸濁させた培地の１／４をプレートに播き、３７℃でインキュベートすることにより、ＨｕＶＥＣの継代培養を行った。 Subculture was performed as follows. First, the EBM-2 medium was removed from the dish in which HuVECs were cultured, and the HuVECs on the dish were washed once with PBS, and the PBS was removed after washing. Next, 1 mL of 0.05% trypsin EDTA (1x) (Gibco) was added to the HuVECs on the dish and incubated at 37°C for 3 minutes. After incubation, the dish was lightly tapped to gently detach the HuVECs from the bottom of the dish. After adding 10 mL of EBM-2 medium containing growth factors to the dish, the HuVECs on the dish were collected in a 15 mL tube. The 15 mL tube was centrifuged at 1000 rpm for 3 minutes at room temperature, the supernatant was removed, and 1 mL of new EBM-2 medium was added to suspend the HuVECs. One-fourth of the medium in which the HuVECs were suspended was plated and incubated at 37°C to subculture the HuVECs.
ベニバナエキスがＨｕＶＥＣの増殖に与える影響の評価
第３～５継代ＨｕＶＥＣを、成長因子を含むＥＢＭ－２培地で培養した。コラーゲンＩでコーティングされた２４ウェルプレート（イワキ社製）に、２×１０４細胞数/ウェルになるように細胞を播き、その２日後、培地を、０．１％のベニバナエキスを含むＥＢＭ－２培地、または含まないＥＢＭ－２培地に交換した。なお、ベニバナエキスの抽出は、公知の方法に準じ、抽出溶媒として１，３ブチレングリコール水溶液を用いて行った。ＨｕＶＥＣの増殖は、アラマーブルー（BIOSOURCE INTERNATIONAL INC社製）を、培地の１０％になるように添加し、３７℃で２時間インキュベート後、その培地を、黒色の９６ウェルプレート（住友ベークライト社製）に２００μＬ／ウェルになるように移し、Ｇｅｎ５マイクロプレートリーダー（励起５５５ｎｍ、発光５９６ｎｍ）を用いてその蛍光を測定することにより評価した。得られた結果に対しては、t-test（P<0.01の場合**、P<0.001の場合***とそれぞれ記載）による統計解析を行った。
Evaluation of the effect of safflower extract on the proliferation of HuVECs The 3rd to 5th passage HuVECs were cultured in EBM-2 medium containing growth factors. The cells were seeded at 2 x 104 cells/well on a collagen I-coated 24-well plate (Iwaki Co., Ltd.), and two days later, the medium was replaced with EBM-2 medium containing 0.1% safflower extract or EBM-2 medium without safflower extract. The safflower extract was extracted using an aqueous 1,3-butylene glycol solution as an extraction solvent according to a known method. HuVEC proliferation was evaluated by adding Alamar Blue (BIOSOURCE INTERNATIONAL INC.) to the medium at 10% of the medium, incubating at 37°C for 2 hours, transferring the medium to a black 96-well plate (Sumitomo Bakelite Co., Ltd.) at 200 μL/well, and measuring the fluorescence using a Gen5 microplate reader (excitation 555 nm, emission 596 nm). Statistical analysis of the results was performed using t-test (P<0.01 is indicated as **, P<0.001 is indicated as ***).
結果を図１および２に示す。ベニバナエキスを添加してから２４時間後および４８時間後のいずれの群においても、ベニバナエキスを添加していないコントロールと比較して、ＨｕＶＥＣの細胞数が有意に増加していることが示された。以上の結果から、ベニバナエキスはＨｕＶＥＣの増殖を促進させることが示された。 The results are shown in Figures 1 and 2. It was shown that the number of HuVEC cells was significantly increased in both groups 24 and 48 hours after the addition of safflower extract, compared to the control group to which no safflower extract was added. These results demonstrate that safflower extract promotes the proliferation of HuVEC.
ベニバナエキスがＨｕＶＥＣおけるＶＥ－カドヘリン発現に与える影響の評価
コラーゲンＩ－Ｃ（クラボウ社製）を０．１ｍＭのＨＣｌで１：１０に希釈した。１ディッシュあたり５００μＬになるように広げ、室温で３０分間インキュベートした。次に、ディッシュを、ＰＢＳで２回洗浄し、成長因子を含むＥＢＭ－２培地で１回洗浄した。１×１０５細胞／ウェルに調製した第３～５継代ＨｕＶＥＣを、コラーゲンＩ－Ｃでコーティングした３５ｃｍガラス底ディッシュ（イワキ社製）に播いた。このディッシュにベニバナエキスを０．１％になるように適用し、３７℃で２４時間、インキュベートした後、ＨｕＶＥＣをＰＢＳで１回洗浄し、４％ＰＦＡ（ナカライテスク社製）で、室温で１０分間固定し、固定したＨｕＶＥＣに０．５％となるようにＴｒｉｔｏｎ-１０×（ナカライテスク社製）を添加して１５分間、室温でインキュベートした後、ＰＢＳ（１×）で３回洗浄し、固定したＨｕＶＥＣにＢＳＡ（ＳＩＧＭＡ社製）を１％になるように添加し、室温で１時間インキュベートした。続いて、１％のＢＳＡで２００倍に希釈した一次抗体、抗ウサギＶＥ－カドヘリン（Cell Signaling Technology社製）を、固定したＨｕＶＥＣに適用し、室温で２時間インキュベートした。一次抗体を適用したＨｕＶＥＣをＰＢＳで３回洗浄した後、１％のＢＳＡで８００倍に希釈した二次抗体、ファロイジン－４８８（Thermo Fisher Scientific社製）および抗ウサギ５６８（Thermo Fisher Scientific社製）を適用し、アルミホイルで覆いながら室温で１時間インキュベートした。二次抗体溶液を除去した後、ＰＢＳで１０００倍に希釈したＨｏｅｃｈｓｔ３３３４２を、二次抗体を適用したＨｕＶＥＣに１０分間適用し、染色した。
Evaluation of the effect of safflower extract on VE-cadherin expression in HuVECs Collagen I-C (Kurabo) was diluted 1:10 with 0.1 mM HCl. 500 μL was spread per dish and incubated at room temperature for 30 minutes. The dishes were then washed twice with PBS and once with EBM-2 medium containing growth factors. HuVECs from passages 3 to 5, prepared at 1×10 5 cells/well, were seeded on 35 cm glass-bottom dishes (Iwaki) coated with collagen I-C. Safflower extract was applied to this dish at 0.1% and incubated at 37°C for 24 hours, after which the HuVECs were washed once with PBS, fixed with 4% PFA (Nacalai Tesque) at room temperature for 10 minutes, Triton-10x (Nacalai Tesque) was added to the fixed HuVECs at 0.5% and incubated at room temperature for 15 minutes, washed three times with PBS (1x), BSA (SIGMA) was added to the fixed HuVECs at 1% and incubated at room temperature for 1 hour. Subsequently, a primary antibody, anti-rabbit VE-cadherin (Cell Signaling Technology), diluted 200-fold with 1% BSA, was applied to the fixed HuVECs and incubated at room temperature for 2 hours. After washing the primary antibody-treated HuVECs three times with PBS, secondary antibodies, phalloidin-488 (Thermo Fisher Scientific) and anti-rabbit 568 (Thermo Fisher Scientific), diluted 800-fold in 1% BSA, were applied and incubated at room temperature for 1 hour while covered with aluminum foil. After removing the secondary antibody solution, Hoechst 33342 diluted 1000-fold in PBS was applied to the secondary antibody-treated HuVECs for 10 minutes for staining.
染色したＨｕＶＥＣを、ＬＳＭ８８０共焦点顕微鏡（ＺＥＩＳＳ社製）の２０倍レンズで観察した（使用レーザー：４８８、５６１、６３３、レーザー出力：４８８ｎｍ(２％)、６３３ｎｍ(６．５％)、４０５ｎｍ(２％)、ゲイン：５５０～６００、Ｚスタック：２０スライス）。ＣＺＩ画像は、さらなる分析のために最大強度の投影ＴＩＦ画像に変換した。 Stained HuVECs were observed under a 20x lens on a ZEISS LSM880 confocal microscope (Lasers used: 488, 561, 633; Laser power: 488 nm (2%), 633 nm (6.5%), 405 nm (2%); Gain: 550-600; Z-stack: 20 slices). CZI images were converted to maximum intensity projection TIF images for further analysis.
結果を図３に示す。ベニバナエキスを添加したＨｕＶＥＣは、ベニバナエキスを添加していないＨｕＶＥＣ（コントロール）と比較して、細胞同士が密に接着していることが示された。 The results are shown in Figure 3. HuVECs to which safflower extract had been added were shown to have tighter cell adhesion compared to HuVECs to which safflower extract had not been added (control).
続いて、ベニバナエキスがＨｕＶＥＣおけるＶＥ－カドヘリン遺伝子発現に与える影響を評価した。まず、第３～５継代ＨｕＶＥＣを、成長因子を含むＥＢＭ－２培地で培養した。コラーゲンでコーティングされた２４ウェルプレート（イワキ社製）に、２×１０４細胞／ウェルで、１ウェルあたり５００μＬの培地になるようにＨｕＶＥＣを播いた。その２日後、培地を０．１％のベニバナエキスを含むＥＢＭ－２培地、またはベニバナエキス含まないＥＢＭ－２培地に交換した。この処理をしたＨｕＶＥＣを、QIAshredder（ＱＩＡＧＥＮ社製）を用いてホモジナイズし、次に、ＲＮｅａｓｙ Ｍｉｎｉキット（ＱＩＡＧＥＮ社製）を用いてＲＮＡ精製をした。精製したｍＲＮＡを逆転写し、リアルタイム定量ＰＣＲは、以下の表１に示すＩＤを有する、Ｔａｑｍａｎ（Thermo Fisher Scientific社製）プライマープローブセットを用い、LightCyclerラピッドサーマルサイクラーシステム（Ｒｏｃｈｅ社製）を用いて、実行した。ＰＣＲの反応条件は、初期変性４８℃で１５秒、変性９５℃で１０分、変性ーアニールー伸長を９５℃で１０分、６０℃で１分を繰り返し回数４０回とした。群のＶＥ－カドヘリン遺伝子の発現レベルをｄｄＣｔ値として得た後、ハウスキーピング遺伝子であるアクチンの発現レベルを参照遺伝子として使用し、各群のＶＥ－カドヘリン遺伝子の発現レベルを補正した。補正した後、ベニバナエキスを添加していない群（コントロール）のＶＥ－カドヘリン遺伝子の発現レベルに対するベニバナエキスを添加した群のＶＥ－カドヘリン遺伝子の相対的発現レベルを計算した。得られた結果に対しては、t-testによる検定（P<0.05の場合*と記載）による統計解析を行った。 Next, the effect of safflower extract on VE-cadherin gene expression in HuVEC was evaluated. First, the 3rd to 5th passage HuVEC were cultured in EBM-2 medium containing growth factors. HuVEC were seeded on a collagen-coated 24-well plate (Iwaki) at 2 x 104 cells/well with 500 μL of medium per well. Two days later, the medium was replaced with EBM-2 medium containing 0.1% safflower extract or EBM-2 medium without safflower extract. The treated HuVEC was homogenized using QIAshredder (QIAGEN), and then RNA was purified using RNeasy Mini kit (QIAGEN). The purified mRNA was reverse transcribed, and real-time quantitative PCR was performed using a Taqman (Thermo Fisher Scientific) primer probe set with the IDs shown in Table 1 below, and a LightCycler rapid thermal cycler system (Roche). The PCR reaction conditions were initial denaturation at 48°C for 15 seconds, denaturation at 95°C for 10 minutes, denaturation-annealing extension at 95°C for 10 minutes, and 60°C for 1 minute, repeated 40 times. After obtaining the expression level of the VE-cadherin gene of each group as a ddCt value, the expression level of the housekeeping gene actin was used as a reference gene to correct the expression level of the VE-cadherin gene of each group. After correction, the relative expression level of the VE-cadherin gene of the group to which safflower extract was added to the group to which safflower extract was not added (control) was calculated. The results obtained were subjected to statistical analysis using t-test (P<0.05 is indicated with *).
結果を図４に示す。ベニバナエキスを添加したＨｕＶＥＣにおいては、コントロールと比較して、有意にＶＥ－カドヘリン遺伝子が上昇していた。以上、図３および４の結果から、ベニバナエキスはＶＥ－カドヘリンの発現を増加させることが示された。 The results are shown in Figure 4. In HuVECs to which safflower extract was added, VE-cadherin gene expression was significantly increased compared to the control. The results of Figures 3 and 4 show that safflower extract increases the expression of VE-cadherin.
ベニバナエキスがＨｕＶＥＣおける細胞内カルシウム濃度に与える影響の評価
第３～５継代ＨｕＶＥＣを、成長因子を含むＥＢＭ－２培地で培養した。コラーゲンIでコーティングされたガラスウェルプレート（イワキ社製）に、１×１０５細胞／ウェルになるようにＨｕＶＥＣを播いた。その２日後、培養液をリンゲル液（１４０ｍＭのＮａＣｌ、５ｍＭのＫＣｌ、１０ｍＭのＨＥＰＥＳ、２ｍＭのＣａＣｌ２、２ｍＭのＭｇＣｌ２、１０ｍＭのＤグルコースを含む溶液）に置換し、蛍光プローブ試薬である、５ｍＭのＦｕｒａ－２－ＡＭ（Thermo Fisher Scientific Japan社製）をリンゲル液中のＨｕＶＥＣに３０分間、３７℃でロードした。このＨｕＶＥＣに、ベニバナエキスを０．１％または０．２％適用し、オリンパスIX83、UPLFLN10xレンズ（オリンパス社製）を用いて細胞内カルシウム濃度の経時的な変化を測定した。
Evaluation of the effect of safflower extract on intracellular calcium concentration in HuVECs The 3rd to 5th passage HuVECs were cultured in EBM-2 medium containing growth factors. HuVECs were seeded at 1 x 105 cells/well on a collagen I-coated glass well plate (Iwaki). Two days later, the culture medium was replaced with Ringer's solution (a solution containing 140 mM NaCl, 5 mM KCl, 10 mM HEPES, 2 mM CaCl2, 2 mM MgCl2, and 10 mM D-glucose), and 5 mM Fura-2-AM (Thermo Fisher Scientific Japan), a fluorescent probe reagent, was loaded onto the HuVECs in Ringer's solution for 30 minutes at 37°C. Safflower extract was applied to the HuVECs at 0.1% or 0.2%, and the time-dependent changes in intracellular calcium concentration were measured using an Olympus IX83 and UPLFLN10x lens (Olympus Corporation).
結果を図５に示す。ベニバナエキスを０．１％または０．２％適用した場合のいずれにおいても、ＨｕＶＥＣの細胞内においてカルシウム濃度が上昇していることが観察された。したがって、ベニバナエキスは細胞内のカルシウム濃度を増加させることが示された。 The results are shown in Figure 5. When safflower extract was applied at either 0.1% or 0.2%, an increase in calcium concentration was observed in the HuVEC cells. This indicates that safflower extract increases calcium concentration in the cells.
Claims (6)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022089908A JP7526758B2 (en) | 2022-06-01 | 2022-06-01 | Composition comprising safflower extract for stabilizing blood vessels in a subject |
| CN202310565837.XA CN117137959A (en) | 2022-06-01 | 2023-05-19 | Composition for stabilizing blood vessel in subject comprising safflower extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022089908A JP7526758B2 (en) | 2022-06-01 | 2022-06-01 | Composition comprising safflower extract for stabilizing blood vessels in a subject |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2023177144A JP2023177144A (en) | 2023-12-13 |
| JP7526758B2 true JP7526758B2 (en) | 2024-08-01 |
Family
ID=88884950
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2022089908A Active JP7526758B2 (en) | 2022-06-01 | 2022-06-01 | Composition comprising safflower extract for stabilizing blood vessels in a subject |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP7526758B2 (en) |
| CN (1) | CN117137959A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007032551A1 (en) | 2005-09-14 | 2007-03-22 | Ajinomoto Co., Inc. | Hemodynamics improving agent |
| JP2013035820A (en) | 2011-05-10 | 2013-02-21 | Maruzen Pharmaceut Co Ltd | Tie2 ACTIVATOR, ANGIOGENESIS SUPPRESSANT, MATURATING AGENT, NORMALIZING AGENT AND STABILIZING AGENT OF BLOOD VESSEL, AND PHARMACEUTICAL COMPOSITION |
| CN104887870A (en) | 2015-06-18 | 2015-09-09 | 王学香 | Anthocyanin oral liquid capable of improving eyesight and preparation method of anthocyanin oral liquid |
-
2022
- 2022-06-01 JP JP2022089908A patent/JP7526758B2/en active Active
-
2023
- 2023-05-19 CN CN202310565837.XA patent/CN117137959A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007032551A1 (en) | 2005-09-14 | 2007-03-22 | Ajinomoto Co., Inc. | Hemodynamics improving agent |
| JP2013035820A (en) | 2011-05-10 | 2013-02-21 | Maruzen Pharmaceut Co Ltd | Tie2 ACTIVATOR, ANGIOGENESIS SUPPRESSANT, MATURATING AGENT, NORMALIZING AGENT AND STABILIZING AGENT OF BLOOD VESSEL, AND PHARMACEUTICAL COMPOSITION |
| CN104887870A (en) | 2015-06-18 | 2015-09-09 | 王学香 | Anthocyanin oral liquid capable of improving eyesight and preparation method of anthocyanin oral liquid |
Non-Patent Citations (4)
| Title |
|---|
| CHEN, Tangting et al.，Hydroxysafflor Yellow A Promotes Angiogenesis via the Angiopoietin 1/ Tie-2 Signaling Pathway，J Vasc Res，2017年，53 (5-6): 245-254，<DOI: 10.1159/000452408> |
| LIU, Ya-Li et al.，Hydroxysafflor yellow A ameliorates lipopolysaccharide-induced acute lung injury in mice via modulating toll-like receptor 4 signaling pathways，International Immunopharmacology，2014年，Volume 23, Issue 2, Pages 649-657，<DOI: 10.1016/j.intimp.2014.10.018> |
| SUN, Yang et al.，Protective cerebrovascular effects of hydroxysafflor yellow A (HSYA) on ischemic stroke，European Journal of Pharmacology，2018年，Volume 818，Pages 604-609，<DOI: 10.1016/j.ejphar.2017.11.033> |
| 福原 茂朋，血管透過性のダイナミックかつ巧妙な制御を可能にするシグナル伝達系，Journal of Japanese Biochemical Society，2017年，89(3):368-376 ，<doi: 10.14952/SEIKAGAKU.2017.890368> |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117137959A (en) | 2023-12-01 |
| JP2023177144A (en) | 2023-12-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pan et al. | SCF promotes dental pulp progenitor migration, neovascularization, and collagen remodeling–potential applications as a homing factor in dental pulp regeneration | |
| JP7564568B2 (en) | Skin Composition | |
| KR20170085010A (en) | Stem cell-derived exosomes containing a high amount of growth factors | |
| JP7034944B2 (en) | Compounds that promote hair growth and / or inhibit or delay hair loss in humans, as well as compositions for such use. | |
| JP5746813B2 (en) | Method for treating, reducing and inhibiting the appearance of skin aging | |
| JP6355222B2 (en) | Composition for suppressing or improving hair loss and whitening and use thereof | |
| JP2016094477A (en) | Maturing, normalizing or stabilizing agent of blood vessel and wrinkle preventing/improving agent | |
| WO2016049867A1 (en) | Composition with exogenous mitochondria as active ingredients, and use thereof and cell repairing method therefor | |
| CN109563132A (en) | Skin regeneration or the peptide of Wound healing and bone regeneration and application thereof | |
| JP4441123B2 (en) | Use of rosmarinus plant extracts in compositions for treating signs of aging skin | |
| EP3656850A1 (en) | Mesenchymal-stem-cell induction agent | |
| JP2006298802A (en) | Ester-bonded chlorogenic acid derivative/alginine having vasorelaxing action and food supplement and cosmetic containing the same | |
| JP5737663B2 (en) | Skin barrier function improver | |
| JP7526758B2 (en) | Composition comprising safflower extract for stabilizing blood vessels in a subject | |
| JP2011115152A (en) | Screening method | |
| US11628133B2 (en) | Uses of the peptide of sequence His-D-Trp-Ala-Trp-d-Phe-Lys-NH2 for reducing or delaying the appearance of cell senescence and signs of skin aging | |
| CN110248641A (en) | Skin regeneration, skin comprising silk gum, common cnidium fruit P.E and Herba Visci extract are releived or wound healing composition | |
| JP5798294B2 (en) | Collagen gel shrinking agent | |
| KR101757734B1 (en) | Cosmetic composition for promoting the stemness of adipose-derived stem cell and proliferating skin cell containing sequoia callus cell extracts | |
| KR101769546B1 (en) | Cosmetic composition comprising daphnin and use thereof | |
| JP2006223144A (en) | Method for screening hair-growing agent with prostaglandin f synthase and/or carbonyl reductase-1 as indicator | |
| KR102372747B1 (en) | Composition for promoting recovery of liver comprising 6-O-trans-feruloyl catalpol | |
| JP6113981B2 (en) | Screening method | |
| KR20170080548A (en) | Cosmetic composition for promoting the stemness of adipose-derived stem cell and proliferating skin cell containing hypericum extracts | |
| KR101766394B1 (en) | Cosmetic composition for promoting the stemness of adipose-derived stem cell and proliferating skin cell containing hypericum extracts |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20240415 |
|
| A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20240415 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20240507 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20240520 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20240628 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20240722 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7526758 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |