Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
  • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
  • A61K8/9783—Angiosperms [Magnoliophyta]
  • A61K8/9789—Magnoliopsida [dicotyledons]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
• • 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)
  • A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
  • A61K8/0216—Solid or semisolid forms
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
  • A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/60—Sugars; Derivatives thereof
  • A61K8/602—Glycosides, e.g. rutin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/63—Steroids; Derivatives thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
  • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
  • A61K2800/74—Biological properties of particular ingredients
  • A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
  • A61K2800/782—Enzyme inhibitors; Enzyme antagonists
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
  • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
  • A61K2800/91—Injection

Definitions

• the present invention relates to a cosmetic composition
• a cosmetic composition comprising extract of Pulsatillae Radix as main ingredient.
• the skin of human being consisted of epidermis and dermis. Nails and hairs are formed in the epidermis and sweat glands exist there too. In dermis there exist nerve nets, blood vessels and sweat glands. Melanin-producing- melanocytes exist in epidermis.
• the starting material for producing melanin is tyrosine, an essential amino acid and the tyrosine is oxidized by tyrosine hydroxylase(TH) into 3,4-dihydroxyphenylalanine(DOPA) and the DOPA is experienced through several reaction steps to be changed into melanin, a black polymer. If the TH-function is congenitally deficient, albinism is appeared. Accordingly, The most important target of development of whitening material in • biosynthetic mechanism of melanin is to find any material which prevents the activity of tyrosine hydroxylase.
• Korean Patent Laid-open Publication No. 2002-0023168 discloses that the extract of Gardeniae fructus has inhibitory effect against tyrosine hydroxylase by 3 times than arbutin in comparative test.
• the said patent specification did not disclose which material in the extract of Gardeniae fructus has such inhibitory action against tyrosine hydroxylase, it could not evaluate whether the extract of Gardeniae fructus has toxic effect on the skin of human being or not.
• materials such as dihydroxybenzene derivatives, retinoid series and steroid hormones had been being used as tyrosine hydroxylase inhibitor.
• the said materials showed to have side effects on the skin and therefore the use of the said materials is restricted.
• the present invention relates to a whitening cosmetic composition comprising extract of Pulsatillae Radix as main ingredient.
• the present invention relates to a whitening cosmetic composition comprising extract of Pulsatillae Radix and one or more ingredient(s) selected from the group consisting ranunculin, deoxypodophyllotoxin and 3-O- ⁇ -L-ramnopyranosyl(l ⁇ 2)-[ ⁇ -D- glucopyranosyl(l— >4)]- ⁇ -L-arabinopyranoside(SB365) extracted and isolated from the extract of Pulsatillae Radix as main ingredients.
• the present invention relates to a whitening cosmetic composition comprising extract of Pulsatillae Radix and extract of bark of Ulmus macrocarpa as main ingredients.
• One object of the present invention is to provide a whitening cosmetic composition comprising the extract of Pulsatillae Radix as main ingredient.
• the other object of the present invention is to provide a whitening cosmetic composition comprising the extract of Pulsatillae Radix and the extract of the bark of the Ulmus macrocarpa as main ingredients.
• Another object of the present invention is to provide a whitening cosmetic composition comprising the extract of Pulsatillae Radix and one or more ingredient(s) selected from the group consisting ranunculin, deoxypodophyllotoxin and 3-O- ⁇ -L- ramnopyranosyl(l— >2)-[ ⁇ -D-glucopyranosyl(l ⁇ 4)]- ⁇ -L-arabinopyranoside(SB365) extracted and isolated from the extract of Pulsatillae Radix as main ingredients.
• Still another object of the present invention is to provide a whitening cosmetic composition
• a whitening cosmetic composition comprising the extract of Pulsatillae Radix as main ingredient and more comprising one or more extracts selected from the group consisting the extract of the bark of the Ulmus macrocarpa, extract of Ginseng Radix and extract of Glycyrrhizae Radix as auxiliary ingredients.
• Still another object of the present invention is to privide a whitening cosmetic composition
• a whitening cosmetic composition comprising the extract of Pulsatillae Radix and one or more ingredient(s) selected from the group consisting ranunculin, deoxypodophyllotoxin and 3-O- ⁇ -L- ramnopyranosyl(l ⁇ 2)-[ ⁇ -D-glucopyranosyl(l ⁇ 4)]- ⁇ -L-arabinopyranoside(SB365) extracted and isolated from the extract of Pulsatillae Radix as main ingredients and more comprising one or more extracts selected from the group consisting the extract of the bark of the Ulmus macrocarpa, the extract of Ginseng Radix and the extract of Glycyrrhizae Radix as auxiliary ingredients.
• Pulsatillae Radix is the root of Pulsatilla koreana, P. cerna and P. chinensis, etc.
• the Pulsatillae Radix has been being used as oriental medicine for treating child bed fever, detoxication, antidiarheal, bactericide, amebicide, fungicide, etc.
• Recently, the Pulsatillae Radix is reported to have antitumor activity and is now under clinical study.
• the bark of Ulmus marocarpa has being been used as oriental medicine for treating various parasites, various kinds of epidermophytid(Oriental Materia Medica, H-Y Hsu et. al., Oriental Healing Arts Institute, pp 749). Therefore, the bark of Ulmus marocarpa is regarded to have protecting effect for the skin.
• Deoxypodophyllotoxin isolated from the extract of Pulsatillae Radix is known compound and has the following structural formula.
• Deoxypodophyllotoxin Ranunculin isolated from the extract of Pulsatillae Radix is known compound and has the following structural formula.
• the said compounds are confirmed to have excellent whitening effects by the present invention.
• the present whitening composition can be formulated by adding conventional vehicles to cream, injection, capsule, tablet, etc., by conventional preparation methods.
• Extracting solvent such as water; lower alkanol e.g. methanol, ethanol, propanol or butanol; methylenechloride; acetone; or mixture thereof can be used in the present invention.
• Fig. 1 indicates the results of chromatogram of PT fraction through Sepadex LH20 column chromatography.
• the Pulsatillae Radix were powdered to obtain powder of 40-100 mesh. A certain volume of the powder was put in an extractor and extraction was carried out with lower alcohol-water mixed solvent. Lower alcohol is selected from the group consisting methanol, ethanol, propanol and butanol. Etanol-water mixed solvent among them is the best selectivity in extraction. In the case of the use of ethanol-water mixed solvent of 50%(v/v), materials having very big polarity and materials of large molecular weight and polymers, etc. were not extracted. Extraction were carried out under the temperature of 15 ° C ⁇ 35 ° C, most preferably 20°C ⁇ 25°C. Extraction time carried out from lhour unit and gradually increased. Extraction time of 3hours is preferable. The extract which Pulsatillae Radix was extracted by using 50% aqueous ethanol solvent under the temperature of 25 ° C for 3 hours is expressed as ET fraction and the ET fraction was used for measuring the whitening effect.
• Example 2 Isolation of materials having whitening effect.
• the whitening effect was confirmed by clinical trial with a group of human beings of the following experimental examples.
• the isolation of materials having whitening effect was carried out by the method of measurement for tyrosinase-inhibition effect which is generally used.
• Low molecular, polar materials were extracted from acetone and the acetone- extract was expressed as PA fraction and the remaining part was expressed as PT fraction.
• PT and PA fractions showed respectably 10% and 73% inhibition effects against tyrosinase.
• the fraction 3 which was obtained by re-fractionation of the PT fraction with Sephadex was refined to obtain an active ingredient which was ascertained as 3-O- ⁇ -L- ramnopyranosyl( 1 ⁇ 2)- [ ⁇ -D-glucopyranosyl( 1 ⁇ 4)] - ⁇ -L-arabinopyranoside(SB365) of which generic name is hederagenin.
• This compound is ascertained to have antitumor activity(Now patent pending).
• This material showed no effect against tyrosinase but showed an excellent whitening effect in clinical trials. So, this material is sure to have whitening effect through another mechanism. It is to be assumed that when relation between saponin derivative and the surface of skin cell is to be thought, the compound hinders the migration of melanosome and prevents melanin to be accumulated on keratinocyte.
• the SPX2 fraction which was obtained by fractionation of the PT fraction by Sephadex showed inhibition of 48% against tyrosinase.
• the Rf value was identified as those of ranunculin.
• the NMR data of the material which was isolated by a known method was the same with those of the ranunculin.
• Ranunculin is broadly spread in plants which belonged to Ranunculaceae and is known to have mitotoxicity (Vonderbank, Pharmazie 5, 21(1950)). Pure ranunculin showed inhibition of 51% against tyrosinase.
• Deoxypodophyllotoxin is spread in various plants including Anthriscus sylvestris Hoffm and shows to have mitotoxicity (Byung-Zun Ahn, Song-Bae Kim, Yong Kim, et al. Use of Deoxypodophyllotoxin as antitumor agent, Korean Patent No. 315,200). It is reported that this material inhibits the formation of blood vessel of endotheliocyte of umbilicus in human being(Yong Kim, Song-Bae Kim, Byung-Zun Ahn, et al., Deoxypodophyllotoxin; the cytotoxic and antiangiogenic component from Pulsatilla koreana Nakai, Planta Medica, 68, 271-274(2002)). This material exhibits inhibition of 38% at 0.03 ⁇ g/ml against tyrosinase. It was difficult to experiment at higher concentration because this material has low water solubility.
• Example 3 50g of dried finely powdered Pulsatilla Radix and 500ml of 50% ethanol were added to an extractor. The mixture was stirred at room temperature for 3 hours and filtered. The filtrate was stored and the procedure was carried out with the remaining residue for 2 times. The combined filtrate was concentrated under reduced pressure and dried to obtain 23g of extract.
• PT fraction having inhibition against tyrosinase and preparation of the PA fraction 20g of the extract obtained in Example 3 was added to 200ml of acetone. The mixture was stirred for 10 minutes to obtain a suspension. The suspension was filtered to obtain solution and residue. The residue was suspended in 200ml of acetone and stirred for
• the PT2 fraction (344mg, 61.4%) was collected from the test tube Nos. 66 ⁇ 91 and main sports are 2.
• the PT3 fraction (61mg, 10.9%) was collected from the test tube Nos. 91 ⁇ 111.
• the PT4 fraction (15.7mg, 2.8%) was collected from the test tube Nos. Il l ⁇ 138.
• the SPX3 fraction and the SPX4 fraction were appeared each one spot on the thin layers and were fractions which were relatively pure. The fractions were shown on the
• the SPX3 fraction was relatively pure fraction and was obtained from white precipitation which the fraction was dissolved in 0.5ml of water and stored to precipitate (PTpur).
• this compound was hydrolyzed by ethanol/sulfuric acid.
• °C-NMR and 'H-NMR data of the hydrolyzed product were compared, it was ascertained that PTpur was hederagenin.
• the hydrolyzed saccharides were ascertained as rhamnose, arabinose and glucose by comparative TLC.
• PTpur is ascertained to be 3-O- ⁇ -L-ramnopyranosyl(l- ⁇ 2)-[ ⁇ -D-glucopyranosyl(l ⁇ 4)]- ⁇ -L- arabinopyranoside(SB365) of which generic name is hederagenin, a saponin already isolated from the Pulsatilla Radix.
• each lOOmg, 400mg, 600mg, 800mg and lOOOmg of the ET fraction were added and mixed uniformly to obtain each preparation PI 00, P400, P600, P800, PlOOOmg.
• Glycyrrhizae Radix as auxiliary ingredients were extracted with 50% ethanol.
• the extract was prepared, filled in vials and lyophilized to obtain injections by conventional preparation method of injections.
• the above ingredients were prepared to obtain a capsule by a conventional preparation method of capsules.
• the above ingredients were prepared and filled in ampoule of 1ml to obtain injection by a conventional preparation method of injection.
• the above ingredients were prepared to obtain a capsule by a conventional preparation method of capsules.
• the above ingredients were prepared to obtain tablet by a conventional preparation method of tablets.
• Whitening effect was carried out with 20 volunteer women. Among them 10 of the women had freckles on their faces and remaining 10 women had liver spots on their faces.
• the same methods were applied to the women of liver spots.
• the groups were divided into G, G'(G group), H, H'(H group), I, I'(I group), J, J'(J group), K, L'(K group) and L, L'(L group).
• the volumes of the composition and the base were each 500mg per 5x5cm 2 .
• the numbers of administrations were 3 times a day. Before administrations, faces were washed. The administrations were continued for 3 weeks.
• PI 00 did not show effects on freckle group A,A'(100mg) or liver spot group G,G'(100mg); P400 showed slightly effective on freckle group
• a combined treatment of the administration of injection and ointment was more immediate effective than ointment treatment on skin without injection.
• the PT and PA fractions inhibited 10% and 73% of activity of tyrosinaseat at 5mg/ml respectively.
• Deoxypodophyllotoxin inhibited 38% of activity of tyrosinase at 0.03 ⁇ g/ml. It was difficult that experiment at higher concentration could not be carried out because the material has very low solubility in water.
• composition lOmg of ranunculin, 50mg of deoxypodophyllotoxin, and 15mg of SB365 were mixed well together with the base (19g) to obtain pharmaceutical composition and the composition was rubbed on the area of liver spots three times a day till the time of curing.
• a and B were cured after lOdays; C was cured after 17days; D was cured after 27days and E was not cured, though the composition was slightly effective.
• a composition from which SB365 which was not effective on tyrosinase was deleted was used.
• the composition were administered to five women (F,G,H,I,K) having liver spots.
• F and G were cured after 26days; I was cured after 35days; and J and K were the level of effectiveness.
• SB365 did not inhibit the activity of tyrosinase but have whitening effect.
• SB 365 acts on melanocyte and interrupts the procedure of the migration of melanin to karatinocyte. An exact mechanism may be disclosed scientifically.
• the combined composition can be anticipated to have merits of beauty arts, such as synergic effect of whitening action, skin-protection through bacteriocidal effect of saponin, skin-elutriation and skin-penetration effect, etc.
• Tyrosinase was purchased from Sigma Company. The enzyme was dissolved in 3, 4-dihydrophenylalanine. Sodium phosphate buffer(0.1M, pH 6.0), a substrate of enzyme and adjusted 1.6mg/ml of concentration to obtain an enzyme solution. Sample was dissolved in sodium phosphate buffer and the undissolved residue was filtered out. 0.7ml of the buffer, 0.2ml of sample and 0.1ml (15.7unit/ml) of the enzyme solution were mixed and after 60seconds, absorbance was measured at 475nm by spectrophotometer.
• S Test tube having enzyme and sample
• B Test tube having sample
• C Test tube having enzyme
• Example 3 B-16 cells originated from mouse melanoma were inoculated to each culture mediums and were cultured under the condition of 37 °C, 5% CO 2 for 5 days. The cells were dispersed with trypsin; centrifuged at l,000rpm for 5minutes; collected and the melanoid degrees were measured macroscopically.
• the composition of the present invention has excellent whitening effect on melanin originated from the mouse melanoma.
• each 2x2 cm 2 parts were chosen ; the chosen parts were washed well with warm water; other parts than the chosen parts were masked with aluminum foil in order for ultraviolet ray to be irradiated only on the chosen parts; and with two FL20SBLB lamp(Toshiba Co., Ltd.) and FL20SE-30 lamp(Toshiba Co., Ltd.) at the same time, ultraviolet ray was irradiated the volume of 0.8x10erg/cm 3 /time/day for successively 3 times.
• a vanishing cream exempt the extract of Pulsatilla Radix was prepared and used as control.
• the cosmetic composition comprising extract of Pulsatilla Radix of the present invention has excellent inhibiting effect against melanin formation.
• the present invention relates to a whitening cosmetic composition
• a whitening cosmetic composition comprising the extract of Pulsatillae Radix and if necessary, one or more ingredient(s) selected from the group consisting ranunculin, deoxypodophyllotoxin and 3-O- ⁇ -L-ramnopyranosyl(l ⁇ 2)-[ ⁇ - D-glucopyranosyl(l ⁇ 4)]- ⁇ -L-arabinopyranoside(SB365) extracted and isolated from the extract of Pulsatillae Radix as main ingredients and if necessary, more comprising one or more extracts selected from the group consisting the extract of the bark of the Ulmus macrocarpa, the extract of Ginseng Radix and the extract of Glycyrrhizae Radix as auxiliary ingredients.
• the present composition has an excellent whitening effect.

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• 2002
  • 2002-09-23 KR KR10-2002-0057653A patent/KR100515206B1/ko not_active IP Right Cessation
• 2003
  • 2003-09-18 JP JP2004538039A patent/JP2006517180A/ja active Pending
  • 2003-09-18 AU AU2003263624A patent/AU2003263624A1/en not_active Abandoned
  • 2003-09-18 WO PCT/KR2003/001905 patent/WO2004026275A1/en active Application Filing
  • 2003-09-18 EP EP03797740A patent/EP1549285A4/en not_active Withdrawn
  • 2003-09-18 CN CNB038226200A patent/CN100408023C/zh not_active Expired - Fee Related
  • 2003-09-18 CA CA002499603A patent/CA2499603A1/en not_active Abandoned

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