JP7466246B1 - Wrinkle improving agent and its manufacturing method - Google Patents
Wrinkle improving agent and its manufacturing method Download PDFInfo
- Publication number
- JP7466246B1 JP7466246B1 JP2023125487A JP2023125487A JP7466246B1 JP 7466246 B1 JP7466246 B1 JP 7466246B1 JP 2023125487 A JP2023125487 A JP 2023125487A JP 2023125487 A JP2023125487 A JP 2023125487A JP 7466246 B1 JP7466246 B1 JP 7466246B1
- Authority
- JP
- Japan
- Prior art keywords
- enzymes
- rice
- cation exchange
- enzyme
- improving agent
- 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
- 230000037303 wrinkles Effects 0.000 title claims description 41
- 238000004519 manufacturing process Methods 0.000 title description 19
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 81
- 235000009566 rice Nutrition 0.000 claims abstract description 81
- 108090000790 Enzymes Proteins 0.000 claims abstract description 61
- 102000004190 Enzymes Human genes 0.000 claims abstract description 61
- 239000007788 liquid Substances 0.000 claims abstract description 43
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 34
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 31
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000003729 cation exchange resin Substances 0.000 claims abstract description 27
- 239000000203 mixture Substances 0.000 claims abstract description 27
- 238000000926 separation method Methods 0.000 claims abstract description 9
- 241000209094 Oryza Species 0.000 claims description 80
- 229940088598 enzyme Drugs 0.000 claims description 60
- 238000000855 fermentation Methods 0.000 claims description 33
- 230000004151 fermentation Effects 0.000 claims description 33
- 239000000284 extract Substances 0.000 claims description 28
- 238000000605 extraction Methods 0.000 claims description 19
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 18
- 230000001476 alcoholic effect Effects 0.000 claims description 13
- 239000004310 lactic acid Substances 0.000 claims description 9
- 235000014655 lactic acid Nutrition 0.000 claims description 9
- 239000007787 solid Substances 0.000 claims description 8
- 108091005804 Peptidases Proteins 0.000 claims description 5
- 102000035195 Peptidases Human genes 0.000 claims description 5
- 230000002615 fibrolytic effect Effects 0.000 claims description 5
- 230000002366 lipolytic effect Effects 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 3
- 238000001179 sorption measurement Methods 0.000 claims description 3
- 230000003625 amylolytic effect Effects 0.000 claims 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 6
- 240000007594 Oryza sativa Species 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 19
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- 238000000354 decomposition reaction Methods 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 12
- 229920002674 hyaluronan Polymers 0.000 description 12
- 210000003491 skin Anatomy 0.000 description 12
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 11
- 229960003160 hyaluronic acid Drugs 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- -1 pH adjusters Substances 0.000 description 11
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 9
- 239000002953 phosphate buffered saline Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 230000001737 promoting effect Effects 0.000 description 8
- 102000008186 Collagen Human genes 0.000 description 7
- 108010035532 Collagen Proteins 0.000 description 7
- 235000021329 brown rice Nutrition 0.000 description 7
- 238000005341 cation exchange Methods 0.000 description 7
- 229920001436 collagen Polymers 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 239000002537 cosmetic Substances 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 210000002469 basement membrane Anatomy 0.000 description 5
- 230000037319 collagen production Effects 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000003960 organic solvent Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 235000011121 sodium hydroxide Nutrition 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 229920002472 Starch Polymers 0.000 description 4
- 239000004480 active ingredient Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000006071 cream Substances 0.000 description 4
- 239000000839 emulsion Substances 0.000 description 4
- 230000035784 germination Effects 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 239000003860 topical agent Substances 0.000 description 4
- 102100026802 72 kDa type IV collagenase Human genes 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 239000004129 EU approved improving agent Substances 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 3
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000003028 enzyme activity measurement method Methods 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 230000004130 lipolysis Effects 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000017854 proteolysis Effects 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 2
- 101000627872 Homo sapiens 72 kDa type IV collagenase Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000003149 assay kit Methods 0.000 description 2
- 239000013040 bath agent Substances 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 239000000919 ceramic Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 239000012139 lysis buffer Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 239000002674 ointment Substances 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 238000002731 protein assay Methods 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000007665 sagging Methods 0.000 description 2
- 238000010025 steaming Methods 0.000 description 2
- 210000000434 stratum corneum Anatomy 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- 101710151806 72 kDa type IV collagenase Proteins 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000519695 Ilex integra Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000002582 Oryza sativa Indica Group Species 0.000 description 1
- 240000008467 Oryza sativa Japonica Group Species 0.000 description 1
- 239000006096 absorbing agent Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000010564 aerobic fermentation Methods 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 230000001153 anti-wrinkle effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000000306 component Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- KIUKXJAPPMFGSW-MNSSHETKSA-N hyaluronan Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)C1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H](C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-MNSSHETKSA-N 0.000 description 1
- 229940099552 hyaluronan Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- JIRXORZYIXSWOB-UHFFFAOYSA-N n-hydroxy-2-[(4-methoxyphenyl)sulfonyl-(2-methylpropyl)amino]acetamide Chemical compound COC1=CC=C(S(=O)(=O)N(CC(C)C)CC(=O)NO)C=C1 JIRXORZYIXSWOB-UHFFFAOYSA-N 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000001121 post-column derivatisation Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 208000017520 skin disease Diseases 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 125000000020 sulfo group Chemical group O=S(=O)([*])O[H] 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
【課題】 従来よりも優れた、新規なシワ改善剤を提供することを課題とする。【解決手段】 米に水を加えたものに対して、酵素を作用させ、アルコール発酵させた後、固液分離にて得た液体部を陽イオン交換樹脂で処理し、該陽イオン交換樹脂に吸着された成分を回収することにより得られたシワ改善剤である。【選択図】 図1[Problem] The objective of the present invention is to provide a new wrinkle-improving agent that is superior to conventional products. [Solution] The wrinkle-improving agent is obtained by adding water to rice, treating the mixture with an enzyme, fermenting it into alcohol, and then treating the liquid portion obtained by solid-liquid separation with a cation exchange resin and recovering the components adsorbed by the cation exchange resin. [Selected Figure] Figure 1
Description
本発明は、シワ改善剤及びその製造方法に関する。 The present invention relates to a wrinkle improving agent and a method for producing the same.
従来から、米を原料とした外用剤が多く提案されている。例えば、特許文献1~6には、様々な用途に関し、米の粉砕物を有効成分として含有する組成物;米の抽出物を有効成分として含有する組成物;前記抽出物が、米に水又は有機溶媒を加えたもの、米を酸又はアルカリで処理したもの、米の加水物に酵素又は麹を作用させたもの、或いは、これらを加熱又は非加熱下でおこなったものである組成物;米を抽出するに際し、その抽出前、抽出と同時又は抽出後に酵素又は麹を作用させたものを有効成分として含有する組成物が開示されている。 Many topical agents made from rice have been proposed. For example, Patent Documents 1 to 6 disclose, for various uses, compositions containing ground rice as an active ingredient; compositions containing rice extracts as an active ingredient; compositions in which the extract is obtained by adding water or an organic solvent to rice, treating rice with acid or alkali, reacting enzymes or koji with hydrated rice, or compositions obtained by carrying out these procedures with or without heating; and compositions containing active ingredients obtained by reacting enzymes or koji before, simultaneously with, or after rice extraction.
本発明は、米を原料とした、シワ改善に特に有効な組成物を提供することを課題とする。 The objective of the present invention is to provide a composition made from rice that is particularly effective in improving wrinkles.
本発明(1)は、米に水を加えたもの又はその抽出物に対して、酵素及び/又は米麹を作用させたもの或いはその抽出物を、アルコール発酵及び/又は乳酸発酵させて発酵物を得、前記発酵物を固液分離して液体部を得、前記液体部を陽イオン交換樹脂に通した後、前記陽イオン交換樹脂に吸着された成分を溶出させることにより得られたことを特徴とする、シワ改善剤である。
本発明(2)は、前記発酵物が、アルコール発酵させたものである、前記発明(1)のシワ改善剤である。
本発明(3)は、米に水を加えたもの又はその抽出物に対して、酵素及び/又は米麹を作用させたもの或いはその抽出物を得る、加水又は抽出工程と、
前記加水又は抽出工程後、アルコール発酵及び/又は乳酸発酵させて発酵物を得る発酵工程と、
前記発酵物を固液分離して液体部を得る固液分離工程と、
前記液体部を陽イオン交換樹脂に通した後、前記陽イオン交換樹脂に吸着された成分を溶出させる吸着・溶出工程と
を有することを特徴とする、シワ改善剤の調製方法である。
本発明(4)は、前記発酵工程での発酵が、アルコール発酵である、前記発明(3)の調製方法である。
The present invention (1) is a wrinkle improving agent obtained by subjecting a product obtained by adding water to rice or an extract thereof to the action of an enzyme and/or rice koji, or an extract thereof, to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, subjecting the fermented product to solid-liquid separation to obtain a liquid portion, passing the liquid portion through a cation exchange resin, and then eluting the components adsorbed to the cation exchange resin.
The present invention (2) relates to the wrinkle improving agent of the above-mentioned invention (1), wherein the fermented product is an alcoholic fermented product.
The present invention (3) provides a method for producing a rice product by adding water to rice or an extract thereof, and then reacting an enzyme and/or rice koji with the resulting product or an extract thereof;
A fermentation step of obtaining a fermented product by alcoholic fermentation and/or lactic acid fermentation after the water addition or extraction step;
A solid-liquid separation step of separating the fermented product into solid and liquid to obtain a liquid portion;
The method for preparing a wrinkle improving agent is characterized by comprising an adsorption/elution step of passing the liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin.
The present invention (4) relates to the preparation method according to the invention (3), wherein the fermentation in the fermentation step is alcoholic fermentation.
本発明によれば、米を原料とした、シワ改善に特に有効な組成物を提供することができる。 The present invention provides a composition made from rice that is particularly effective in improving wrinkles.
本発明の一形態である本形態は、好適には、米に水を加えたもの又はその抽出物に対して、酵素及び/又は米麹を作用させたもの或いはその抽出物を、アルコール発酵及び/又は乳酸発酵させて発酵物を得、前記発酵物を固液分離して液体部を得、前記液体部を陽イオン交換樹脂に通した後、前記陽イオン交換樹脂に吸着された成分を溶出させることにより得られたことを特徴とする、シワ改善剤である。 This embodiment, which is one aspect of the present invention, is preferably a wrinkle improving agent obtained by subjecting rice to water or an extract thereof to the action of enzymes and/or rice koji, or an extract thereof, to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, separating the fermented product into solid and liquid to obtain a liquid portion, passing the liquid portion through a cation exchange resin, and then eluting the components adsorbed to the cation exchange resin.
≪シワ改善剤の製造方法≫
本形態に係るシワ改善剤は、例えば、(第一工程)米に水を加えたもの又はその抽出物を得る工程、(第二工程)第一工程で得られたものに対して、酵素及び/又は米麹を作用させる(又はそれを抽出する)工程、(第三工程)第二工程で得られたものに対して、アルコール発酵及び/又は乳酸発酵させて発酵物を得る工程、(第四工程)第三工程で得られた発酵物を固液分離して液体部を得る工程、(第五工程)第四工程で得られた液体部を陽イオン交換樹脂に通した後、該陽イオン交換樹脂に吸着された成分を溶出させる陽イオン吸着・溶出工程、により得られる。以下、本形態に係るシワ改善剤の製造方法を詳述する。
<Production method of wrinkle improvement agent>
The wrinkle improving agent according to the present embodiment can be obtained, for example, by the following steps: (first step) a step of adding water to rice or obtaining an extract thereof, (second step) a step of applying an enzyme and/or rice koji to the product obtained in the first step (or extracting it), (third step) a step of subjecting the product obtained in the second step to alcoholic fermentation and/or lactic acid fermentation to obtain a fermented product, (fourth step) a step of separating the fermented product obtained in the third step from solid to liquid to obtain a liquid portion, and (fifth step) a cation adsorption/elution step of passing the liquid portion obtained in the fourth step through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin. The manufacturing method of the wrinkle improving agent according to the present embodiment will be described in detail below.
<第一工程>
第一工程は、米に水を加えたもの又はその抽出物を得る工程である。ここで、「米」とは、白米、玄米及び発芽させた米といった米だけでなく、白糠及び赤糠等といった米の一部も含む概念である。但し、白米部分を必須的に含むもの、即ち、白米、玄米、発芽させた米、白糠を用いることが好適である。ここで、白米及び玄米に関しては、ジャポニカ、インディカ米を問わず、うるち米、及び餅米等の玄米及び白米を指し、品種、種類は問わない。また、白糠及び赤糠とは、一般に、精白時に出てくる92%以上の赤糠や92%以下の白糠を指すが、両者が混合したものを使用してもよい。尚、上記の原料は、浸漬、蒸煮、焙煎(砂焙り、網焙り、熱風焙煎等全てを指す)、蒸煮焙煎、凍結乾燥等の表面変性、UV照射等の光変性、パットライス等の加圧焙煎、揚げる等の原料処理をしてもよい。白米、玄米及び発芽させた米は、そのまま用いてもよく、粉砕して用いてもよい。白米、玄米及び発芽させた米を粉砕して粉体化するには、粉砕機又は精米機を用い一般的な方法で行なえばよい。
<First step>
The first step is a step of adding water to rice or obtaining an extract thereof. Here, "rice" is a concept including not only white rice, brown rice, and germinated rice, but also parts of rice such as white bran and red bran. However, it is preferable to use those that essentially contain a white rice portion, that is, white rice, brown rice, germinated rice, and white bran. Here, white rice and brown rice refer to brown rice and white rice such as non-glutinous rice and mochi rice, regardless of whether they are Japonica or Indica rice, regardless of the variety or type. In addition, white bran and red bran generally refer to 92% or more red bran and 92% or less white bran that are produced during polishing, but a mixture of both may be used. The above raw materials may be subjected to raw material treatment such as soaking, steaming, roasting (including sand roasting, net roasting, hot air roasting, etc.), surface modification such as steaming roasting, freeze-drying, photo-modification such as UV irradiation, pressurized roasting such as pat rice, and frying. Polished rice, brown rice and germinated rice may be used as is or may be crushed before use. Polished rice, brown rice and germinated rice may be crushed into powder by a general method using a grinder or rice polisher.
発芽させた米を製造する場合、胚芽のついた米を水に浸漬又は水を噴霧して発芽させる。発芽させる時の温度は10~50℃である。但し、発芽さえすれば、温度及び時間は問わない。また、発芽中に水が腐敗する危険性がある場合は、腐敗しないように水を取り替えるか、何らかの防腐を行うのが好ましい。ここで、発芽とは、発芽する直前から発芽したものまで全てを指す。この発芽させた米をよく洗浄して用いる。この際、乾燥して用いてもよい。米を抽出、或いは酵素分解又は麹を作用させる場合、原料の米を粉砕して顆粒或いは粉体化すると、表面積が大きくなるため効率がよくなる。粉砕しなくてもよいが、この場合には、米組織の分解及び抽出に長時間を要する。 When producing germinated rice, rice with germ is soaked in water or sprayed with water to germinate. The temperature during germination is 10 to 50°C. However, as long as germination occurs, the temperature and time are not important. If there is a risk of the water spoiling during germination, it is preferable to replace the water or use some kind of preservative to prevent spoilage. Germinated rice here refers to everything from just before germination to when the rice has germinated. This germinated rice is washed thoroughly before use. It may also be dried before use. When extracting rice or subjecting it to enzymatic decomposition or koji treatment, it is more efficient to crush the raw rice into granules or powder because the surface area is larger. It is not necessary to crush the rice, but in this case it takes a long time to decompose and extract the rice tissue.
次に、「抽出」とは、米を液体媒体で処理することを指す。ここで、液体媒体での処理としては、水抽出、酸抽出、アルカリ抽出、有機溶媒抽出等を挙げることができる。まず、米を水抽出する場合、抽出温度は、高温が効率的であるが、低温でも十分に抽出を行うことができる。但し、40℃以下の低温の場合は、pHを酸性又はアルカリ性にするか、防腐剤又はアルコールを加えて、米が腐敗しないように処理することが望ましい。抽出時間は、有効成分さえ抽出できれば、長くても短くてもよく、抽出温度により定めればよい。また、抽出は、加圧下又は常圧下で行っても、減圧下で行ってもよい。水抽出の場合、最も問題になるのは糊化現象である。糊状になれば、抽出効率が悪くなるばかりでなく、実作業においては困難を極める。これを防ぐためには、アミラーゼを加えて反応させるか、塩酸等で酸性にして澱粉を切ってやればよく、この方法を用いることにより、十分に解決でき、実用上も全く問題はない。また、酸分解抽出又はアルカリ分解抽出を行う場合、必要により中和、脱塩を行う。有機溶媒で抽出する場合も、米はなるべく微粉砕又は粉体化して抽出することが望ましい。有機溶媒はアルコール、アセトン、n-へキサン、酢酸エチル、メタノール等の一般的な有機溶媒でよいが、人体に対して有害なものは抽出後、溶媒を完全に除去する必要があるので安全なものがよい。 Next, "extraction" refers to treating rice with a liquid medium. Examples of treatment with a liquid medium include water extraction, acid extraction, alkali extraction, and organic solvent extraction. First, when extracting rice with water, a high extraction temperature is effective, but low temperatures are also sufficient for extraction. However, when the temperature is low, such as 40°C or less, it is desirable to treat the rice so that it does not spoil by making the pH acidic or alkaline, or adding a preservative or alcohol. The extraction time can be long or short as long as the active ingredients can be extracted, and can be determined according to the extraction temperature. Extraction can be performed under pressure, normal pressure, or reduced pressure. In the case of water extraction, the biggest problem is gelatinization. If the rice becomes pasty, not only will the extraction efficiency decrease, but it will also be extremely difficult to carry out in practical use. To prevent this, amylase can be added to cause a reaction, or the rice can be acidified with hydrochloric acid or the like to cut off the starch. This method can be used to fully solve the problem and is completely problem-free in practice. When performing acid decomposition extraction or alkali decomposition extraction, neutralization and desalting are performed as necessary. When extracting with an organic solvent, it is desirable to pulverize or powder the rice as finely as possible before extraction. The organic solvent may be any common organic solvent such as alcohol, acetone, n-hexane, ethyl acetate, or methanol, but it is better to use a safe one, as those that are harmful to the human body must be completely removed after extraction.
<第二工程>
第二工程は、第一工程で得られたもの(米に水を加えたもの又はその抽出物)に対して、酵素及び/又は米麹を作用させる(又はそれを抽出する)工程である。ここで、酵素は、澱粉分解酵素(液化酵素、糖化酵素)、蛋白分解酵素、脂肪分解酵素、繊維分解酵素、リグニン分解酵素、ペクチン分解酵素等、米に働く酵素である。また、酵素は、1種でも2種以上を組み合わせてもよい(例えば、液化酵素と糖化酵素との組み合わせ)。また、米麹は、特に限定されない。即ち、麹菌の種類並びに米の品種及び種類は特に問わない。尚、第一工程と第二工程とは、同時に実施しても、異なったタイミングで行ってもよい(即ち、抽出の前、抽出と同時又は抽出の後に、酵素分解及び米麹を作用させてもよい)。
<Second step>
The second step is a step of allowing enzymes and/or rice koji to act on (or extract from) the product obtained in the first step (rice plus water or an extract thereof). Here, the enzymes are enzymes that act on rice, such as starch-decomposing enzymes (liquefying enzymes, saccharifying enzymes), proteolytic enzymes, lipolytic enzymes, fibrolytic enzymes, lignin-decomposing enzymes, and pectin-decomposing enzymes. The enzymes may be one type or a combination of two or more types (for example, a combination of liquefying enzymes and saccharifying enzymes). The rice koji is not particularly limited. That is, the type of koji mold and the variety and type of rice are not particularly important. The first and second steps may be performed simultaneously or at different times (that is, enzymatic decomposition and rice koji may be applied before, simultaneously with, or after extraction).
<第三工程>
第三工程は、第二工程で得られたものを、アルコール発酵及び/又は乳酸発酵する工程である。ここで、発酵形態としては、もろみ発酵及び液体発酵のいずれでもよく、もろみ発酵が好適である。これらの発酵を2回以上繰り返す、又は異なる発酵法を組み合わせてもよい。尚、酵母による通気発酵、アルコール沈殿等を行なって除糖してもよい。
<Third process>
The third step is a step of alcoholic fermentation and/or lactic acid fermentation of the product obtained in the second step. The fermentation form may be either moromi fermentation or liquid fermentation, with moromi fermentation being preferred. These fermentations may be repeated two or more times, or different fermentation methods may be combined. Sugar may be removed by aerobic fermentation using yeast, alcohol precipitation, or the like.
<第四工程>
第四工程は、第三工程で得られた発酵物を固液分離して液体部を得る工程である。ここで、固液分離の手法は、特に限定されず、例えば、圧搾や濾過である。
<Fourth step>
The fourth step is a step of obtaining a liquid portion by performing solid-liquid separation on the fermented product obtained in the third step. The method of solid-liquid separation is not particularly limited, and may be, for example, squeezing or filtration.
<第五工程>
第五工程は、第四工程で得られた液体部を陽イオン交換樹脂に通し、該陽イオン交換樹脂に吸着された成分を取得する陽イオン交換処理工程である。ここで、陽イオン交換樹脂は、特に限定されず、強酸性(スルホ基等)であっても弱酸性(カルボキシル基等)であってもよい。また、陽イオン交換樹脂に吸着された成分の回収方法も特に限定されないが、好適には、苛性ソーダで溶出させることが好適である。
<Fifth step>
The fifth step is a cation exchange treatment step in which the liquid portion obtained in the fourth step is passed through a cation exchange resin to obtain the components adsorbed on the cation exchange resin. The cation exchange resin is not particularly limited, and may be strongly acidic (sulfo group, etc.) or weakly acidic (carboxyl group, etc.). The method for recovering the components adsorbed on the cation exchange resin is not particularly limited, but is preferably eluted with caustic soda.
<他の工程等>
本形態に係るシワ改善剤の製造方法は、上述した第一工程~第五工程以外の工程を有していてもよい。特に、該製造方法は、第五工程(陽イオン交換処理工程)の前及び/又は後に煮沸させる工程を有することが好適である。
<Other processes, etc.>
The manufacturing method of the wrinkle improving agent according to the present embodiment may include steps other than the above-mentioned steps 1 to 5. In particular, the manufacturing method preferably includes a boiling step before and/or after the fifth step (cation exchange treatment step).
<工程の実施タイミング等>
本形態に係るシワ改善剤の製造方法は、基本的には、上述した第一工程~第五工程の順で実施され得る。ここで、上述した第一工程~第五工程及び他の工程に関しては、ある工程が完了した後に次の工程を実施してもよく、ある工程が完了する前に次の工程も並行して実施してもよく、ある工程と次の工程を同時に実施してもよい。例えば、上述の第一工程と第二工程において、酵素により水抽出物を得ることを想定したとき、米に水を入れた後に酵素を添加した場合には、第一工程(米に水を添加)の後に第二工程(米に水を入れたものに酵素を添加)を実施するケースに該当し、酵素を含有する水に米を入れる場合には、第一工程と第二工程を同時に実施するケースに該当する。また、本形態に係るシワ改善剤の製造方法は、上述した第一工程~第五工程以外の工程を含んでいてもよい。
<Timing of process implementation, etc.>
The manufacturing method of the wrinkle improving agent according to the present embodiment can basically be carried out in the order of the first step to the fifth step described above. Here, with regard to the first step to the fifth step and other steps described above, the next step may be carried out after the completion of a certain step, the next step may be carried out in parallel before the completion of a certain step, or a certain step and a certain step may be carried out simultaneously. For example, in the above-mentioned first and second steps, when it is assumed that a water extract is obtained by an enzyme, if the enzyme is added after adding water to rice, this corresponds to a case in which the first step (adding water to rice) is followed by the second step (adding enzyme to rice in water), and if rice is added to water containing an enzyme, this corresponds to a case in which the first step and the second step are carried out simultaneously. In addition, the manufacturing method of the wrinkle improving agent according to the present embodiment may include steps other than the above-mentioned first step to the fifth step.
≪シワ改善剤の性質≫
<コラーゲン産生促進作用>
本形態に係るシワ改善剤は、前述のように、特定の液体部を陽イオン交換樹脂に通した後該陽イオン交換樹脂に吸着された成分を溶出させることで得られたことを特徴とするところ、好適には、陽イオン交換樹脂を通さない該特定の液体部と比較し、好適には、5%以上、7.5%以上、10%以上、12.5%以上、15%以上、17.5%以上、20%以上、25%以上、30%以上、35%以上、40%以上のコラーゲン産生促進作用を有する(上限は特に限定されず、例えば500%)。
<Properties of anti-wrinkle agents>
<Promotes collagen production>
As described above, the wrinkle improving agent of this embodiment is characterized in that it is obtained by passing a specific liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin, and preferably has a collagen production promoting effect of 5% or more, 7.5% or more, 10% or more, 12.5% or more, 15% or more, 17.5% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more (the upper limit is not particularly limited, for example 500%), compared to the specific liquid portion that has not been passed through a cation exchange resin.
<基底膜分解抑制作用>
本形態に係るシワ改善剤は、好適には、基底膜分解抑制作用を有する。ここで、基底膜分解抑制作用は、例えば、マトリックスメタロプロテアーゼ(例えば、MMP2)の抑制作用を有することを指す。
<Inhibitory effect on basement membrane degradation>
The wrinkle improving agent according to the present embodiment preferably has an effect of inhibiting basement membrane decomposition. Here, the effect of inhibiting basement membrane decomposition refers to, for example, the effect of inhibiting matrix metalloproteinase (e.g., MMP2).
<ヒアルロン酸産生促進作用>
本形態に係るシワ改善剤は、前述のように、特定の液体部を陽イオン交換樹脂に通した後該陽イオン交換樹脂に吸着された成分を溶出させることで得られたことを特徴とするところ、好適には、陽イオン交換樹脂を通さない該特定の液体部と比較し、好適には、2.5%以上、5%以上、7.5%以上、10%以上、12.5%以上のヒアルロン酸産生促進作用を有する(上限は特に限定されず、例えば500%)。
<Promoting hyaluronic acid production>
As described above, the wrinkle improving agent of this embodiment is characterized in that it is obtained by passing a specific liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin, and preferably has a hyaluronic acid production promoting effect of 2.5% or more, 5% or more, 7.5% or more, 10% or more, or 12.5% or more (the upper limit is not particularly limited, for example 500%) compared to the specific liquid portion that has not been passed through a cation exchange resin.
<NMF産生促進作用>
本形態に係るシワ改善剤は、前述のように、特定の液体部を陽イオン交換樹脂に通した後該陽イオン交換樹脂に吸着された成分を溶出させることで得られたことを特徴とするところ、好適には、陽イオン交換樹脂を通さない該特定の液体部と比較し、好適には、2.5%以上、7.5%以上、10%以上、12.5%以上、15%以上、17.5%以上、20%以上、22.5%以上、25%以上のNMF産生促進作用を有する(上限は特に限定されず、例えば500%)。
<NMF production promoting effect>
As described above, the wrinkle improving agent of this embodiment is characterized in that it is obtained by passing a specific liquid portion through a cation exchange resin and then eluting the components adsorbed to the cation exchange resin, and preferably has an NMF production promoting effect of 2.5% or more, 7.5% or more, 10% or more, 12.5% or more, 15% or more, 17.5% or more, 20% or more, 22.5% or more, or 25% or more (the upper limit is not particularly limited, for example 500%), compared to the specific liquid portion that has not been passed through a cation exchange resin.
≪シワ改善剤の用途≫
本形態に係るシワ改善剤は、該組成物そのまま又は外用剤への添加成分として有用である。特に、コラーゲン産生促進作用、基底膜分解抑制作用、ヒアルロン酸産生促進作用、NMF産生促進作用を有するシワ改善剤として有用である。ここで、本形態に係るシワ改善剤は、各種添加剤を含有していてもよい。例えば、通常医薬品・医薬部外品・化粧品(例えば、皮膚化粧料、浴用剤、洗浄剤)に添加される成分、具体的には、賦形剤、結合剤、崩壊剤、滑沢剤、着香剤、着色剤、溶解補助剤、懸濁化剤、乳化剤、コーティング剤、保湿剤、洗浄剤、紫外線吸収剤、薬剤を挙げることができる。具体的には、例えば、化粧料として用いる場合には、化粧料で汎用される水性成分、油性成分、粉末、界面活性剤、油剤、pH調整剤、防腐剤、アルコール、酸化防止剤、増粘剤、色素、香料、動物抽出液、植物抽出液等を必要に応じて適宜配合することにより調整する。
<Applications of wrinkle improvement agents>
The wrinkle improving agent according to the present embodiment is useful as the composition itself or as an added component to external preparations. In particular, it is useful as a wrinkle improving agent having collagen production promoting action, basement membrane decomposition inhibiting action, hyaluronic acid production promoting action, and NMF production promoting action. Here, the wrinkle improving agent according to the present embodiment may contain various additives. For example, the components usually added to medicines, quasi-drugs, and cosmetics (for example, skin cosmetics, bath agents, and detergents), specifically, excipients, binders, disintegrants, lubricants, fragrances, colorants, solubilizing agents, suspending agents, emulsifiers, coating agents, moisturizing agents, detergents, ultraviolet absorbers, and drugs can be mentioned. Specifically, for example, when used as a cosmetic, the composition is adjusted by appropriately blending aqueous components, oily components, powders, surfactants, oils, pH adjusters, preservatives, alcohols, antioxidants, thickeners, pigments, fragrances, animal extracts, plant extracts, and the like that are commonly used in cosmetics as necessary.
(適用対象)
本形態に係るシワ改善剤の適用対象は、特に限定されず、例えば、中高年や高齢者のみならず、皮膚疾患者や若者(例えば、下記のシワ防止の用途)をも挙げることができる。また、シワも特に限定されず、例えば、小ジワ(例えば乾燥)、表情ジワ、たるみジワ(ほうれい線)、大ジワ、老齢によるたるみ、痩せたことによるハリのロストを挙げることができる。また、本明細書にいう「改善」は、シワが無くなるか減ることのみならず、シワの防止や抑制をも包含する概念である。
(Applicable subjects)
The wrinkle improving agent of this embodiment is not particularly limited to the target, and can be used not only for middle-aged and elderly people, but also for skin disease sufferers and young people (for example, for the purpose of preventing wrinkles as described below).Wrinkles are also not particularly limited, and can be used for small wrinkles (for example, dryness), facial wrinkles, sagging wrinkles (laugh lines), large wrinkles, sagging due to aging, and loss of firmness due to weight loss.In addition, "improvement" as used in this specification is a concept that includes not only the disappearance or reduction of wrinkles, but also the prevention and suppression of wrinkles.
(適用方法)
本形態に係るシワ改善剤は、皮膚に適用(例えば、塗布、吹付等)して使用する(典型的には、経皮外用剤)。例えば、皮膚外用剤、洗浄剤及び浴用剤等の医薬品、医薬部外品及び化粧品として用い得る。具体的には、ローション等の可溶化系(液状皮膚外用剤、化粧水等)、乳液等の乳化系(乳液状ファンデーション、O/W乳化型美容液等)、粉末・顆粒系(入浴化粧料等)、クリーム系(O/W乳化型クリーム、ハンドクリーム、メイクアップベースクリーム等)、軟膏系(O/W型乳剤性軟膏等)、気体系(エアゾール等)等、様々な剤型で提供可能である。より具体的には、塗布等の他、皮下注射、マイクロニードルパッチ、パップ剤、シートマスクも挙げることができる。
(Method of applying)
The wrinkle improving agent according to the present embodiment is used by applying (e.g., coating, spraying, etc.) to the skin (typically, a transdermal topical agent). For example, it can be used as medicines, quasi-drugs, and cosmetics such as skin topical agents, cleansers, and bath agents. Specifically, it can be provided in various dosage forms such as solubilized systems such as lotions (liquid skin topical agents, skin lotions, etc.), emulsion systems such as emulsions (emulsion foundations, O/W emulsion-type beauty essences, etc.), powders and granules (bath cosmetics, etc.), creams (O/W emulsion-type creams, hand creams, makeup base creams, etc.), ointments (O/W emulsion ointments, etc.), and gas systems (aerosols, etc.). More specifically, in addition to coating, subcutaneous injections, microneedle patches, poultices, and sheet masks can also be mentioned.
(適用量)
本形態に係るシワ改善剤を皮膚に適用する場合、適用する組成物の全質量を基準として、該組成物中に前記シワ改善剤を0.0001質量%以上含有したものを用いることが好適であり、0.001質量%以上含有したものを用いることがより好適であり、0.01質量%以上含有したものを用いることが更に好適である。尚、上限値は特に限定されず、例えば100質量%である。
(Amount applied)
When the wrinkle improving agent according to the present embodiment is applied to the skin, it is preferable to use a composition containing 0.0001% by mass or more of the wrinkle improving agent based on the total mass of the composition to be applied, more preferable to use a composition containing 0.001% by mass or more, and even more preferable to use a composition containing 0.01% by mass or more. The upper limit is not particularly limited, and is, for example, 100% by mass.
以下、実施例を参照しながらより具体的に本発明を説明するが、本発明は該実施例に限定されるものではない。 The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.
≪シワ改善剤の調製≫
<実施例1>
白米500gに澱粉分解酵素2g、蛋白分解酵素2g、脂肪分解酵素2g、繊維分解酵素2gと水1500mlを加え、50℃で20時間放置した。その後、徐々に温度を上げていき、5分間煮沸抽出した後、濾過して、その後、酵母を加え、20日間アルコール発酵を行った。濾過器を用いて固液分離を行い85℃で30分加熱した後、陽イオン交換カラム(三菱ケミカル DIAION SK104H)に通し水で洗浄した後、カラム吸着成分を苛性ソーダで溶出させ中和し0.4Lを得た。
<Preparation of wrinkle improvement agent>
Example 1
2g of starch decomposition enzyme, 2g of proteolysis enzyme, 2g of lipolysis enzyme, 2g of fiber decomposition enzyme and 1500ml of water were added to 500g of polished rice and left to stand at 50°C for 20 hours. After that, the temperature was gradually raised, boiled and extracted for 5 minutes, filtered, yeast was added and alcohol fermentation was carried out for 20 days. After solid-liquid separation using a filter and heating at 85°C for 30 minutes, the mixture was passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the column adsorbed components were eluted with caustic soda and neutralized to obtain 0.4L.
<実施例2>
白米500gに米麹125gと水1500mlを加え、55℃で48時間放置した。その後、酵母を加え、20日間アルコール発酵を行った。濾過器を用いて固液分離を行い85℃で30分加熱した後、陽イオン交換カラム(三菱ケミカル DIAION SK104H)に通し水で洗浄した後、カラム吸着成分を苛性ソーダで溶出させ中和し0.4Lを得た。
Example 2
125g of rice koji and 1500ml of water were added to 500g of polished rice, and the mixture was left at 55°C for 48 hours. Yeast was then added, and alcoholic fermentation was carried out for 20 days. The mixture was separated into solid and liquid using a filter, heated at 85°C for 30 minutes, passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the components adsorbed on the column were eluted with caustic soda and neutralized to obtain 0.4L.
<実施例3>
玄米を粉砕機にかけ、粉砕物500gを得た。この粉砕物に米麹125gと水1500mlを加え、55℃で48時間放置した。その後、酵母を加え、20日間アルコール発酵を行った。濾過器を用いて固液分離を行い85℃で30分加熱した後、陽イオン交換カラム(三菱ケミカル DIAION SK104H)に通し水で洗浄した後、カラム吸着成分を苛性ソーダで溶出させ中和し0.4Lを得た。
Example 3
Brown rice was put into a grinder to obtain 500 g of ground material. 125 g of rice koji and 1500 ml of water were added to the ground material and left at 55°C for 48 hours. Yeast was then added and alcoholic fermentation was carried out for 20 days. After solid-liquid separation using a filter and heating at 85°C for 30 minutes, the mixture was passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the column adsorbed components were eluted with caustic soda and neutralized to obtain 0.4 L.
<実施例4>
白米500gに澱粉分解酵素2g、蛋白分解酵素2g、脂肪分解酵素2g、繊維分解酵素2gと水1500mlを加え、50℃で20時間放置した。その後、徐々に温度を上げていき、5分間煮沸抽出した後、濾過して、その後、乳酸菌を加え、5日間乳酸発酵を行った。濾過器を用いて固液分離を行い85℃で30分加熱した後、陽イオン交換カラム(三菱ケミカル DIAION SK104H)に通し水で洗浄した後、カラム吸着成分を苛性ソーダで溶出させ中和し0.4Lを得た。
Example 4
2g of starch decomposition enzyme, 2g of proteolysis enzyme, 2g of lipolysis enzyme, 2g of fiber decomposition enzyme and 1500ml of water were added to 500g of polished rice, and the mixture was left at 50°C for 20 hours. The temperature was then gradually increased, and the mixture was boiled for 5 minutes for extraction, filtered, and then lactic acid bacteria was added and lactic acid fermentation was carried out for 5 days. The mixture was separated into solid and liquid using a filter, heated at 85°C for 30 minutes, passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the components adsorbed on the column were eluted with caustic soda and neutralized to obtain 0.4L.
<実施例5>
白米500gに米麹125gと水1500mlを加え、55℃で48時間放置した。その後、酵母を加え、20日間アルコール発酵を行った。濾過器を用いて固液分離を行い85℃で30分加熱した後、陽イオン交換カラム(三菱ケミカル DIAION SK104H)に通し水で洗浄した後、カラム吸着成分を0.2Nの塩化ナトリウムで溶出させ0.4Lを得た。
Example 5
125g of rice koji and 1500ml of water were added to 500g of polished rice, and the mixture was left at 55°C for 48 hours. Yeast was then added, and alcoholic fermentation was carried out for 20 days. The mixture was separated into solid and liquid using a filter, heated at 85°C for 30 minutes, passed through a cation exchange column (Mitsubishi Chemical DIAION SK104H) and washed with water, and the components adsorbed on the column were eluted with 0.2N sodium chloride to obtain 0.4L.
<比較例>
白米500gに澱粉分解酵素2g、蛋白分解酵素2g、脂肪分解酵素2g、繊維分解酵素2gと水1500mlを加え、50℃で20時間放置した。その後、徐々に温度を上げていき、5分間煮沸抽出した後、濾過して、その後、酵母を加え、 20日間アルコール発酵を行った。濾過器を用いて固液分離を行い85℃で30分加熱した後、減圧濃縮により0.4Lを得た。
Comparative Example
2g of starch decomposition enzyme, 2g of proteolysis enzyme, 2g of lipolysis enzyme, 2g of fiber decomposition enzyme and 1500ml of water were added to 500g of polished rice and left to stand at 50℃ for 20 hours. After that, the temperature was gradually raised, boiled for 5 minutes, filtered, yeast was added and alcohol fermentation was carried out for 20 days. Solid-liquid separation was carried out using a filter, and the mixture was heated at 85℃ for 30 minutes, and then concentrated under reduced pressure to obtain 0.4L.
≪試験例≫
<試験例1.コラーゲン産生試験評価>
(試薬類)
・細胞:ヒト真皮由来正常線維芽細胞(クラボウ)
・培地:10%FBS(Fetal Bovine Serum)+1%L-グルタミン-ペニシリン-ストレプトマイシン含有DMEM培地(Sigma-Aldrich)
・PBS(Sigma-Aldrich)
・L-アスコルビン酸2-リン酸エステル3ナトリウム(富士フイルム和光純薬)
・ペプシン(Sigma-Aldrich)
・酢酸、特級(富士フイルム和光純薬)
・Sircol Soluble Collagen Assay Kit(biocolor)
(試験方法)
ヒト由来真皮線維芽細胞を組織培養用12well plateに0.8×105cells/wellとなるように播種し、DMEMを加え、コンフルエントまで培養した。各種調整した試料が1.25%になるよう調製したL-アスコルビン酸2-リン酸エステル3ナトリウム50μmol/L含有DMEMに交換し、計21日間培養した。培養は全て37℃・5%CO2環境下で行った。培養終了後、PBSで洗浄し、1mg/mLペプシン含有0.5mol/L酢酸水溶液でコラーゲンを抽出した。抽出液を市販のSircol Soluble Collagen Assay Kitに従い規定通り吸光プレートリーダーを用いてコラーゲン量を求めた。試験は独立して3回実施し、1wellあたりのコラーゲン量と、Controlを100%とした時の相対値と、を算出して比較した。表1は、実施例1~5及び比較例に係る米抽出物について本試験を実施した際における、1wellあたりのコラーゲン量である。表2は、実施例1~5及び比較例に係る米抽出物について本試験を実施した際における、Controlを100%とした時のコラーゲン量の相対値である(図1は、該相対値を図示)。
<Test Example>
<Test Example 1. Collagen Production Test Evaluation>
(Reagents)
Cells: Human dermis-derived normal fibroblasts (Kuraray)
Culture medium: 10% FBS (Fetal Bovine Serum) + 1% L-glutamine-penicillin-streptomycin-containing DMEM medium (Sigma-Aldrich)
PBS (Sigma-Aldrich)
・L-Ascorbic acid 2-phosphate trisodium ester (FUJIFILM Wako Pure Chemical Industries, Ltd.)
Pepsin (Sigma-Aldrich)
・Acetic acid, special grade (FUJIFILM Wako Pure Chemical Industries, Ltd.)
・Sircol Soluble Collagen Assay Kit (biocolor)
(Test method)
Human dermal fibroblasts were seeded in a 12-well plate for tissue culture at 0.8×10 5 cells/well, and DMEM was added and cultured until confluent. The medium was replaced with DMEM containing 50 μmol/L of trisodium L-ascorbic acid 2-phosphate ester, which was prepared so that the various samples were 1.25%, and cultured for a total of 21 days. All cultures were performed in a 37°C, 5% CO 2 environment. After the culture was completed, the cells were washed with PBS, and collagen was extracted with a 0.5 mol/L aqueous acetic acid solution containing 1 mg/mL pepsin. The amount of collagen in the extract was determined using a spectrophotometric plate reader according to the commercially available Sircol Soluble Collagen Assay Kit. The test was performed three times independently, and the amount of collagen per well and the relative value when the control was taken as 100% were calculated and compared. Table 1 shows the amount of collagen per well when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example. Table 2 shows the relative values of the amount of collagen when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example, with the control being taken as 100% (Figure 1 shows the relative values).
<試験例2.基底膜分解試験>
(試薬類)
・MMP酵素活性測定キット:MMP-2 Fluorometric Drug Discovery Kit,RED(Enzo)
(試験方法)
市販のヒトMMP酵素活性測定キットを用いて規定通りの方法により測定した。Control、陽性対照であるNNGH、各種濃度に希釈した試料をそれぞれ添加した時の酵素活性を蛍光検出プレートリーダーにより、励起波長535nm、検出波長590nmの条件で測定し、Controlを100%とした際の相対値を算出した。試料濃度と得られた相対値から酵素活性曲線を作成し、酵素活性を50%阻害する濃度、IC50(%)を算出した。表3は、実施例1~5及び比較例に係る米抽出物についての、in vitro MMP2酵素活性測定の結果である。尚、比較例は酵素活性阻害作用がなく算出できなかった。
Test Example 2: Basement membrane decomposition test
(Reagents)
・MMP enzyme activity measurement kit: MMP-2 Fluorometric Drug Discovery Kit, RED (Enzo)
(Test method)
The enzyme activity was measured by a method specified using a commercially available human MMP enzyme activity measurement kit. The enzyme activity when the control, the positive control NNGH, and the samples diluted to various concentrations were added was measured using a fluorescence detection plate reader under conditions of an excitation wavelength of 535 nm and a detection wavelength of 590 nm, and the relative value was calculated when the control was set as 100%. An enzyme activity curve was created from the sample concentration and the obtained relative value, and the concentration at which the enzyme activity was inhibited by 50%, IC50 (%), was calculated. Table 3 shows the results of in vitro MMP2 enzyme activity measurement for the rice extracts according to Examples 1 to 5 and Comparative Example. The Comparative Example had no enzyme activity inhibitory effect and could not be calculated.
<試験例3.ヒアルロン酸産生試験>
(試薬類)
・細胞:ヒト3次元表皮皮膚モデルEPI-200(クラボウ)
・培地:EPI-200専用培地(クラボウ)
・PBS(Sigma-Aldrich)
・セラミックス製粉砕ボール,φ2mm(東レ)
・Quantikine ELISA Hyaluronan(R&D Systems)
・DC プロテインアッセイ(バイオラッド)
(試験方法)
ヒト3次元表皮皮膚モデル(φ8mm)を専用培地で1日間培養後、各種試料150μLを角質層側に添加、毎日、培地交換、試料交換しながら2日間培養した。培養は全て、37℃・5%CO2環境下で行った。培養終了後、PBSで洗浄し、皮膚モデルを取り出し、PBSを加えて、ビーズクラッシャーにより、細胞を破砕し、抽出液のヒアルロン酸量は、市販の専用キットに基づいて規定通りの試験方法で吸光プレートリーダーにより求めた。タンパク質量は市販の専用キットに基づいて吸光プレートリーダーにより求めた。各ヒアルロン酸量をタンパク質量で除し、単位タンパク質あたりのヒアルロン酸量を算出した。また、陰性対照の単位タンパク質あたりのヒアルロン酸量を100%とした時の相対値を算出した。表4は、実施例1~5及び比較例に係る米抽出物について本試験を実施した際における、単位タンパク質あたりのヒアルロン酸量である。表5は、実施例1~5及び比較例に係る米抽出物について本試験を実施した際における、陰性対照の単位タンパク質あたりのヒアルロン酸量を100%とした時の相対値である(図2は、該相対値を図示)。
<Test Example 3. Hyaluronic acid production test>
(Reagents)
・Cells: Human 3D epidermal skin model EPI-200 (Kurabo)
Culture medium: EPI-200 dedicated culture medium (Kurabo)
PBS (Sigma-Aldrich)
・Ceramic grinding ball, φ2mm (Toray)
・Quantikine ELISA Hyaluronan (R&D Systems)
・DC Protein Assay (Bio-Rad)
(Test method)
After culturing a human three-dimensional epidermal skin model (φ8 mm) in a dedicated medium for one day, 150 μL of each sample was added to the stratum corneum side, and the culture was cultured for two days while changing the medium and sample every day. All cultures were performed in a 37°C, 5% CO2 environment. After the culture was completed, the skin model was washed with PBS, removed, PBS was added, and the cells were crushed using a bead crusher. The amount of hyaluronic acid in the extract was determined using an absorption plate reader according to the prescribed test method based on a commercially available dedicated kit. The amount of protein was determined using an absorption plate reader based on a commercially available dedicated kit. The amount of hyaluronic acid was divided by the amount of protein to calculate the amount of hyaluronic acid per unit protein. In addition, a relative value was calculated when the amount of hyaluronic acid per unit protein of the negative control was set to 100%. Table 4 shows the amount of hyaluronic acid per unit protein when this test was performed on the rice extracts according to Examples 1 to 5 and Comparative Example. Table 5 shows the relative values when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example, with the amount of hyaluronic acid per unit protein of the negative control taken as 100% (Figure 2 shows the relative values).
<試験例4.NMF産生試験>
(試薬類)
・細胞:ヒト3次元表皮皮膚モデルEPI-200(クラボウ)
・培地:EPI-200専用培地(クラボウ)
・PBS(Phosphate Buffered Saline)(Sigma-Aldrich)
・セラミックス製粉砕ボール,φ2mm(東レ)
・RIPA Lysis Buffer system(コスモバイオ)
・アミノ酸混合標準液(Shimadzu)
・DC プロテインアッセイ(バイオラッド)
(試験方法)
ヒト3次元表皮皮膚モデル(φ8mm)を専用培地で1日間培養後、各種試料150μLを角質層側に添加、2日間培養した。培養は全て、37℃・5%CO2環境下で行った。培養終了後、PBSで洗浄し、皮膚モデルを取り出し、PBSを加えて、ビーズクラッシャーにより、破砕した。破砕液RIPA Lysis Bufferで溶解し、NMF定量、タンパク質定量に用いた。タンパク質量は市販の専用キットに基づいて吸光プレートリーダーにより求めた。NMF量は高速液体クロマトグラフィー用いて、オルトフタルアルデヒドポストカラム誘導体化法により求めた。得られたNMF量をタンパク質量で除し、単位タンパク質あたりのNMF量を算出した。また、陰性対照の単位タンパク質あたりのNMF量を100%とした時の相対値を算出した。表6は、実施例1~5及び比較例に係る米抽出物について本試験を実施した際における、単位タンパク質あたりのNMF量である。表7は、実施例1~5及び比較例に係る米抽出物について本試験を実施した際における、陰性対照の単位タンパク質あたりのNMF量を100%とした時の相対値である(図3は、該相対値を図示)。
<Test Example 4. NMF production test>
(Reagents)
・Cells: Human 3D epidermal skin model EPI-200 (Kurabo)
Culture medium: EPI-200 dedicated culture medium (Kurabo)
Phosphate Buffered Saline (PBS) (Sigma-Aldrich)
・Ceramic grinding ball, φ2mm (Toray)
・RIPA Lysis Buffer system (Cosmobio)
Amino acid mixed standard solution (Shimadzu)
・DC Protein Assay (Bio-Rad)
(Test method)
After culturing a human three-dimensional epidermal skin model (φ8 mm) in a dedicated medium for one day, 150 μL of each sample was added to the stratum corneum side and cultured for two days. All culturing was performed in a 37°C, 5% CO2 environment. After the end of the culture, the skin model was washed with PBS, removed, PBS was added, and the model was crushed using a bead crusher. The crushing liquid was dissolved in RIPA Lysis Buffer and used for NMF quantification and protein quantification. The protein amount was determined using a spectrophotometric plate reader based on a commercially available dedicated kit. The NMF amount was determined by ortho-phthalaldehyde post-column derivatization using high performance liquid chromatography. The obtained NMF amount was divided by the protein amount to calculate the NMF amount per unit protein. In addition, a relative value was calculated when the NMF amount per unit protein of the negative control was set to 100%. Table 6 shows the NMF amount per unit protein when this test was performed on the rice extracts according to Examples 1 to 5 and Comparative Example. Table 7 shows the relative values when this test was performed on the rice extracts of Examples 1 to 5 and the Comparative Example, with the NMF amount per unit protein of the negative control taken as 100% (Figure 3 shows the relative values).
Claims (6)
前記加水又は抽出工程後、アルコール発酵及び/又は乳酸発酵させて発酵物を得る発酵工程と、
前記発酵物を固液分離して液体部を得る固液分離工程と、
前記液体部を陽イオン交換樹脂に通した後、前記陽イオン交換樹脂に吸着された成分を溶出させる吸着・溶出工程と
を有することを特徴とする、シワ改善剤の調製方法。 a water addition or extraction step in which at least one enzyme selected from amylolytic enzymes, proteolytic enzymes, lipolytic enzymes and fibrolytic enzymes (excluding a combination of amylolytic enzymes, proteolytic enzymes, lipolytic enzymes and fibrolytic enzymes) or said enzymes and rice koji are allowed to act on the product obtained by adding water to rice or an extract thereof, or an extract thereof is obtained;
A fermentation step of obtaining a fermented product by alcoholic fermentation and/or lactic acid fermentation after the water addition or extraction step;
A solid-liquid separation step of separating the fermented product into solid and liquid to obtain a liquid portion;
and an adsorption/elution step of passing the liquid portion through a cation exchange resin and then eluting the components adsorbed by the cation exchange resin.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2023125487A JP7466246B1 (en) | 2023-04-11 | 2023-08-01 | Wrinkle improving agent and its manufacturing method |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2023064487A JP7330587B1 (en) | 2023-04-11 | 2023-04-11 | Anti-wrinkle agent and method for producing the same |
JP2023125487A JP7466246B1 (en) | 2023-04-11 | 2023-08-01 | Wrinkle improving agent and its manufacturing method |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2023064487A Division JP7330587B1 (en) | 2023-04-11 | 2023-04-11 | Anti-wrinkle agent and method for producing the same |
Publications (1)
Publication Number | Publication Date |
---|---|
JP7466246B1 true JP7466246B1 (en) | 2024-04-12 |
Family
ID=87577185
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2023064487A Active JP7330587B1 (en) | 2023-04-11 | 2023-04-11 | Anti-wrinkle agent and method for producing the same |
JP2023125487A Active JP7466246B1 (en) | 2023-04-11 | 2023-08-01 | Wrinkle improving agent and its manufacturing method |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2023064487A Active JP7330587B1 (en) | 2023-04-11 | 2023-04-11 | Anti-wrinkle agent and method for producing the same |
Country Status (1)
Country | Link |
---|---|
JP (2) | JP7330587B1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP7330587B1 (en) | 2023-04-11 | 2023-08-22 | 株式会社創研 | Anti-wrinkle agent and method for producing the same |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2001253807A (en) | 2000-01-05 | 2001-09-18 | Kamaya:Kk | Pack agent |
WO2007148737A1 (en) | 2006-06-22 | 2007-12-27 | Toyo Boseki Kabushiki Kaisha | Method of preparing plant extract, plant extract and use of the same |
JP2008194040A (en) | 2007-02-12 | 2008-08-28 | Chungbuk Soju Co Ltd | Method for producing alcoholic beverage using cultured root of mountain ginseng |
JP7330587B1 (en) | 2023-04-11 | 2023-08-22 | 株式会社創研 | Anti-wrinkle agent and method for producing the same |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH06104605B2 (en) * | 1986-12-16 | 1994-12-21 | 鐘紡株式会社 | Alcohol-containing cosmetics |
-
2023
- 2023-04-11 JP JP2023064487A patent/JP7330587B1/en active Active
- 2023-08-01 JP JP2023125487A patent/JP7466246B1/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2001253807A (en) | 2000-01-05 | 2001-09-18 | Kamaya:Kk | Pack agent |
WO2007148737A1 (en) | 2006-06-22 | 2007-12-27 | Toyo Boseki Kabushiki Kaisha | Method of preparing plant extract, plant extract and use of the same |
JP2008194040A (en) | 2007-02-12 | 2008-08-28 | Chungbuk Soju Co Ltd | Method for producing alcoholic beverage using cultured root of mountain ginseng |
JP7330587B1 (en) | 2023-04-11 | 2023-08-22 | 株式会社創研 | Anti-wrinkle agent and method for producing the same |
Also Published As
Publication number | Publication date |
---|---|
JP7330587B1 (en) | 2023-08-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP2313082B1 (en) | Cosmetic composition for anti-aging of the skin comprising phaseolus radiatus seed extracts by fermentation and enzyme treatment | |
US8545905B2 (en) | Whitening cosmetic composition containing green tea extract | |
KR101207557B1 (en) | Cosmetic Composition Comprising the Extract of Liriodendron tulipifera as Active Ingredient | |
KR20040057908A (en) | Cosmetics | |
JP7248875B2 (en) | Skin topical composition | |
JP7466246B1 (en) | Wrinkle improving agent and its manufacturing method | |
CN111053716A (en) | Application of Fuzhuan tea extract in preparation of skin conditioning product | |
JP2004250372A (en) | Skin aging inhibitor/improver and/or rough skin inhibitor/improver kit | |
TW201808324A (en) | Skin external agent for skin whitening comprising an extract of fermented wheat germ | |
CN114262667A (en) | Novel aureobasidium pullulans strain and application thereof | |
KR100757175B1 (en) | Skin compositions for external application for improving wrinkle and enhancing skin elasticity comprising kaempferol from green tea | |
JP2016074647A (en) | Rice fermented products by lactic acid bacteria having moisture retention effect | |
TWI388343B (en) | Cosmetic composition for anti-aging of the skin comprising phaseolus radiatus extract of fermentation-enzyme treatment | |
JP7489688B2 (en) | Skin preparations | |
KR20090098705A (en) | Use of a brassocattleya marcella koss orchid extract as an active agent to prevent or delay the appearance of signs of cutaneous aging | |
CN116617137A (en) | Ginseng seed extract and preparation method thereof | |
KR20210060801A (en) | Cosmetic Compositions for Anti-aging Comprising Complex Fermented Product of Plants | |
KR20110121775A (en) | Cosmetic composition comprising the extract of thalictrum rochebrunianum as active ingredient | |
KR101775485B1 (en) | Cosmetic composition containing fermentative extract of Spatholobus suberectus Dunn as active ingredient | |
JP6827691B2 (en) | Cosmetics | |
JP2005001995A (en) | Cosmetic | |
KR20110122448A (en) | Cosmetic composition comprising the extract of filipendula formosa as active ingredient | |
JP2004026766A (en) | Skin care preparation for external use | |
JP7349116B2 (en) | External skin preparations and food and drink products | |
KR102620811B1 (en) | Red centella asiatica fermented with monascus and its use |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20231127 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20231127 |
|
A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20231127 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20231219 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20240117 |
|
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: 20240318 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20240326 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7466246 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |