WO2000056278A1 - Utilisation de la vitamine c ou analogues pour stimuler la synthese de cellules de la peau - Google Patents
Utilisation de la vitamine c ou analogues pour stimuler la synthese de cellules de la peau Download PDFInfo
- Publication number
- WO2000056278A1 WO2000056278A1 PCT/FR2000/000561 FR0000561W WO0056278A1 WO 2000056278 A1 WO2000056278 A1 WO 2000056278A1 FR 0000561 W FR0000561 W FR 0000561W WO 0056278 A1 WO0056278 A1 WO 0056278A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- skin
- ascorbic acid
- analogs
- differentiation
- composition
- Prior art date
Links
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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/676—Ascorbic acid, i.e. vitamin C
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
-
- 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
-
- 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
- A61K2800/70—Biological properties of the composition as a whole
-
- 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
- A61Q19/08—Anti-ageing preparations
Definitions
- the invention relates to a method for increasing the rate of differentiation and / or proliferation of skin fibroblasts and / or increasing the rate of differentiation of skin keratinocytes by applying to the skin a composition comprising an effective amount of ascorbic acid or at least one of its analogs. It also relates to a process for stimulating the synthesis of cutaneous vimentin by applying to the skin a composition comprising an effective amount of ascorbic acid or of at least one of its analogs. It further relates to a process for stimulating the synthesis of cutaneous keratin by applying to the skin a composition comprising an effective amount of ascorbic acid or of at least one of its analogs.
- Human skin is made up of two compartments, namely a surface compartment, the epidermis, and a deep compartment, the dermis.
- the natural human epidermis is mainly composed of three types of cells which are the keratinocytes, very majority, the melanocytes and the Langerhans cells. Each of these cell types contributes by its own functions to the essential role played in the body by the skin, in particular the role of protecting the body from external aggressions (climate, ultraviolet rays, tobacco, ...), called "function fence". Poor renewal of these cells and more particularly of keratinocytes, which is observed in particular with age, results in poor protection of the skin, the skin then having a dry and / or dull appearance.
- the dermis provides the epidermis with solid support. It is also its nourishing element. It mainly consists of fibroblasts and an extracellular matrix itself composed mainly of collagen, elastin and a substance, known as the fundamental substance, components synthesized by the fibroblast. There are also leukocytes, mast cells or tissue macrophages. It is also crossed by blood vessels and nerve fibers.
- Vimentin fibers are found in an important way in the dermis, since they correspond to the intermediate filament of fibroblasts. These vimentin fibers are also present in melanocytes and in Langerhans cells of the epidermis, they can also be present in keratinocytes when these are in a hyperproliferative state.
- Keratins are the intermediate filaments of epithelial cells, such as keratinocytes in the skin. Thus, there are four types of keratin in the epidermis, including keratin 10, called K10, which is specific to the state of differentiation of keratinocytes.
- One of the aims of the present invention is therefore to increase the rate of differentiation and / or proliferation of fibroblasts in the skin and / or to increase the rate of differentiation of keratinocytes in the skin so as to combat extrinsic aggressions, whether they either physical or chemical, which damage the skin, in particular by reducing its function barrier, and against skin aging, whether chronobiological or photo-induced.
- ascorbic acid applied topically to the skin increases the rate of differentiation and / or proliferation of skin fibroblasts and / or increases the rate of differentiation of keratinocytes in the skin by applying to the skin.
- a composition comprising an effective amount of ascorbic acid or at least one of its analogs.
- Ascorbic acid is known to stimulate the synthesis of collagen, preventing, as a co-factor, the auto-inactivation of the enzymes lysine- and proline-hydroxylases and increasing the synthesis of mRNA of procollagens .
- Ascorbic acid (or vitamin C) is also known to stimulate the synthesis of elastin in the skin.
- US patents 5801192, US 4983382 and EP 0717983 We can also cite an article entitled "Pola to incorporate vitamin C in new cosmetics line for skin care" of the Japan economy Journal of June 5, 1984 (page 15) .
- a subject of the invention is therefore the use of an effective amount of ascorbic acid or one of its analogs in a composition or in the preparation of a composition intended to be applied to the skin to increase the rate of differentiation and / or proliferation of skin fibroblasts and / or increase the rate of differentiation of skin keratinocytes.
- a third object of the invention is the use of an effective amount of ascorbic acid or one of its analogs in a composition or in the preparation of a composition intended to be applied to the skin to stimulate the synthesis of cutaneous vimentin.
- a fourth object of the invention is the use of an effective amount of ascorbic acid or one of its analogs in a composition or in the preparation of a composition intended to be applied to the skin to stimulate the synthesis of cutaneous keratin 10.
- the invention further relates to a method for increasing the rate of differentiation and / or proliferation of fibroblasts of the skin in a person having an abnormally low rate of differentiation and / or proliferation of fibroblasts, comprising topical application to the skin with an effective amount of ascorbic acid or one of its analogs.
- Another subject of the invention is a method for increasing the rate of differentiation of keratinocytes in the skin in a person having an abnormally low rate of differentiation of keratinocytes, comprising the topical application to the skin of an effective amount of ascorbic acid or one of its analogs.
- ascorbic acid applied topically to the skin makes it possible to increase the synthesis of mRNA of vimentin and thus to increase the rate of synthesis of vimentin.
- ascorbic acid applied topically to the skin increases the synthesis of keratin 10 mRNA and thus increases the rate of synthesis of keratin 10.
- proteins, intermediate filaments of skin cells are therefore representative of the proliferating and / or differentiating state of skin cells, more particularly cells of the dermis and epidermis. More particularly, vimentin, which is the intermediate filament of fibroblasts, is representative of the proliferating and / or differentiating state of fibroblasts and keratin 10 is representative of the differentiating state of keratinocytes.
- the ratio of the synthesis of mRNAs of vimentin to that of keratin 10 due to the topical application of ascorbic acid is comparable to that without topical application of ascorbic acid. This indicates that the skin condition, after topical application of ascorbic acid, is maintained in a normal state (without, for example, hyper-proliferation or -differentiation of one dermal or epidermal compartment relative to the other) .
- the analogues of ascorbic acid are, more particularly, its salts, such as in particular sodium ascorbate, magnesium or sodium ascorbylphosphate, its esters, such as in particular its acetic, propionic or palmitic esters, or its sugars , such as in particular glycosilated ascorbic acid.
- Ascorbic acid is usually in L-form because it is usually extracted from natural products.
- the effective amount of ascorbic acid or its analogs which can be used according to the invention is of course that which is necessary to obtain the expected effects according to the invention.
- this quantity preferably represents from 0.001% to 20% of the total weight of the composition, preferably from 0.1% to 15% of the total weight of the composition. composition and advantageously from 3% to 10% of the total weight of the composition.
- composition of the invention is used for a time sufficient to obtain the expected effects according to the invention.
- this duration can be at least 15 days, but can also be more than 4 weeks, or even more than 8 weeks.
- composition of the invention intended for topical application contains a physiologically acceptable medium, that is to say compatible with the skin including the scalp, mucous membranes and / or the eyes and may in particular constitute a cosmetic or dermatological composition. .
- This composition can be in all the galenical forms normally used in the cosmetic and dermatological fields, and it can in particular be in the form of an aqueous solution possibly gelled, of a dispersion of the lotion type possibly biphasic, of an emulsion obtained by dispersion of a fatty phase in an aqueous phase (O / W) or vice versa (W / O), or of a triple emulsion (W / O / W or O / W / O) or of a vesicular type dispersion ionic and / or non-ionic.
- These compositions are prepared according to the usual methods.
- composition of the invention can constitute, for example, a lotion, a gel, a cream or a milk, and for example a lotion or a cleansing or cleansing milk, a shampoo or a shower gel.
- composition L-ascorbic acid 5.00%
- Biopsies of these treated surfaces are then carried out.
- Biopsies are ground under liquid nitrogen in a Mikrodismembrator S (Braun).
- the powder obtained is collected in the teflon capsule with 2 ml of lysis solution (guanidine isothiocyanate 5M, mercaptoethanol, 0.1 M, lauryisucosyl of Na 0.017M, citrate Na 0.025M, pH7, antifoam 3 ⁇ l / ml).
- the suspension is transferred to a tube which is stirred at temperature room for 15 minutes.
- the lysate is deposited on the surface of a 1.4 ml cushion of 5.7 M cesium chloride, 0.1 M EDTA, pH 7 in a 3.8 ml polyallomer tube for the rotor SW60 (Beckman L70M ultracentrifuge ). Ultracentrifugation at 35,000 RPM is carried out for 18 hours at 20 ° C. The pellet is rinsed with absolute ethanol, centrifuged at 13,000 RPM, 4 ° C, 10 minutes and dissolved in 100 ⁇ l of distilled water.
- RNA harvested from biopsies is estimated by the optical density of the solution at 260 nm and then measured by amplifying the 28S ribosomal RNA by RT-PCR.
- the measurement of the specific mRNAs is carried out by quantitative RT-PCR on aliquots of the same dilution of total RNA, stored at -80 ° C. until their use.
- the specific oligonucleotide primers of the genes studied have 24 bases, have an A - T% close to 50% and are chosen from two different exons in order to avoid amplification of possible traces of DNA present in the samples.
- the optimal amplification conditions (temperature and number of cycles) were determined for each of the genes studied, taking into account their level of expression in the skin. RT-PCR is carried out using the Gen amp Amp rTth kit from Perkin Elmer or the Titam kit from Boehringer.
- Each RT-PCR reaction is carried out in the presence of a known number of copies of a synthetic RNA created in the laboratory containing the sequences of the oligonucleotide primers specific for the mRNAs of interest and the amplification product of which has a molecular size allowing discriminate it from endogenous mRNA.
- This multistandard makes it possible to control and calculate the yield of the reverse transcription and of the amplification reaction.
- the amplification products are analyzed by electrophoresis in polyacrylamide gel followed by staining with CyberGreen. The intensity of the fluorescent signals is measured using a Fluoro S Multilmager. The results are corrected for the yield of RT-PCR and expressed in arbitrary units per unit of ribosomal 28 S RNA.
- the A / P ratio is significantly greater than 1 with a probability greater than 95% for a value of t> 1.83 and a probability greater than 99% for a value of t> 2.82.
- the total quantity of RNA purified from biopsies is first evaluated by measuring the optical density at 260 nm and their quality estimated by measuring the OD ratio 260/290 nm.
- RNA was obtained from each of the biopsies (between 2.1 and 6.3 ⁇ g) with a satisfactory degree of purity (D.O. report 260/280).
- the concentration of total RNA is brought by dilution to a calculated value of 4 nanograms per ⁇ l. This process makes it possible to carry out the reactions of reverse transcription and amplification on similar amounts of total RNA for all samples.
- the amount of total RNA present in the diluted solution is quantitatively determined by measurement of the 28S ribosomal RNA, carried out in tripiicate. This same RNA solution will be used for all measurements of specific RNAs whose results are expressed per unit of 28S RNA.
- the vimentin / keratin 10 ratio is detailed in Table 3.
- VIM / K10 subject Active Placebo a 7.22 6.05 b 6.88 [14.10] c 5.27 5.63 d 4.53 2.68 e 4.82 4.18 f 2.65 4.31 g 3.59 6.34 h 3.77 5.36 i 3.21 4.48 j 4.19 4.93
- biopsies contain a proportion of mRNA of keratin 10 and of vimentin, comparable on the side treated with ascorbic acid and placebo.
- results also indicate that the biopsies were performed uniformly in the different individuals. The fa-placebo sample is outside the norm.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Toxicology (AREA)
- Cosmetics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2000606185A JP2002541074A (ja) | 1999-03-24 | 2000-03-07 | 皮膚細胞の分化及び/または増殖のレベル増大のためのビタミンc類似物の使用 |
CA002364608A CA2364608A1 (fr) | 1999-03-24 | 2000-03-07 | Utilisation de la vitamine c ou analogues pour augmenter le taux differenciation et/ou de proliferation des cellules de la peau |
AU31726/00A AU3172600A (en) | 1999-03-24 | 2000-03-07 | Use of vitamin c or the like for stimulating skin cell synthesis |
EP00909442A EP1165033A1 (fr) | 1999-03-24 | 2000-03-07 | Utilisation de la vitamine c ou analogues pour stimuler la synthese des cellules de la peau |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR9903675A FR2791259B1 (fr) | 1999-03-24 | 1999-03-24 | Utilisation de la vitamine c ou analogues pour augmenter le taux de differenciation et/ou de proliferation des cellules de la peau |
FR99/03675 | 1999-03-24 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2000056278A1 true WO2000056278A1 (fr) | 2000-09-28 |
Family
ID=9543584
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/FR2000/000561 WO2000056278A1 (fr) | 1999-03-24 | 2000-03-07 | Utilisation de la vitamine c ou analogues pour stimuler la synthese de cellules de la peau |
Country Status (9)
Country | Link |
---|---|
EP (1) | EP1165033A1 (fr) |
JP (1) | JP2002541074A (fr) |
AR (1) | AR018230A1 (fr) |
AU (1) | AU3172600A (fr) |
CA (1) | CA2364608A1 (fr) |
FR (1) | FR2791259B1 (fr) |
TW (1) | TWI225410B (fr) |
UY (1) | UY26063A1 (fr) |
WO (1) | WO2000056278A1 (fr) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE10050274A1 (de) * | 2000-10-09 | 2002-04-18 | Henkel Kgaa | Verfahren zur in vitro Bestimmung der Hautalterung |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP4080946B2 (ja) * | 2003-05-28 | 2008-04-23 | 株式会社クラレ | 皮膚外用剤 |
JP2008105985A (ja) * | 2006-10-25 | 2008-05-08 | Nicca Chemical Co Ltd | ヒアルロン酸産生促進剤、皮膚外用剤、浴用剤及び飲食物 |
JP2008105983A (ja) * | 2006-10-25 | 2008-05-08 | Nicca Chemical Co Ltd | 線維芽細胞増殖促進剤、皮膚外用剤、浴用剤及び飲食物 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS6281307A (ja) * | 1985-10-04 | 1987-04-14 | Kanebo Ltd | 皮膚化粧料 |
FR2612775A1 (fr) * | 1987-03-25 | 1988-09-30 | Thorel Jean | Compositions pour la prevention et le traitement de la degenerescence cutanee |
WO1991018614A1 (fr) * | 1988-11-14 | 1991-12-12 | Bioderm, Inc. | Procede de traitement de la peau vieillissante ou abimee par la lumiere |
WO1995003028A1 (fr) * | 1993-07-23 | 1995-02-02 | Morris Herstein | Composition cosmetique stimulant le renouvellement de la peau, avec effet prolonge d'elimination de l'irritation |
FR2737971A1 (fr) * | 1995-08-25 | 1997-02-28 | Lvmh Rech | Utilisation de la vitamine c ou de ses derives ou analogues pour stimuler la synthese de l'elastine cutanee |
-
1999
- 1999-03-24 FR FR9903675A patent/FR2791259B1/fr not_active Expired - Fee Related
-
2000
- 2000-03-07 EP EP00909442A patent/EP1165033A1/fr not_active Ceased
- 2000-03-07 WO PCT/FR2000/000561 patent/WO2000056278A1/fr not_active Application Discontinuation
- 2000-03-07 JP JP2000606185A patent/JP2002541074A/ja active Pending
- 2000-03-07 CA CA002364608A patent/CA2364608A1/fr not_active Abandoned
- 2000-03-07 AU AU31726/00A patent/AU3172600A/en not_active Abandoned
- 2000-03-14 UY UY26063A patent/UY26063A1/es not_active Application Discontinuation
- 2000-03-20 TW TW089105043A patent/TWI225410B/zh not_active IP Right Cessation
- 2000-03-22 AR ARP000101265A patent/AR018230A1/es unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS6281307A (ja) * | 1985-10-04 | 1987-04-14 | Kanebo Ltd | 皮膚化粧料 |
FR2612775A1 (fr) * | 1987-03-25 | 1988-09-30 | Thorel Jean | Compositions pour la prevention et le traitement de la degenerescence cutanee |
WO1991018614A1 (fr) * | 1988-11-14 | 1991-12-12 | Bioderm, Inc. | Procede de traitement de la peau vieillissante ou abimee par la lumiere |
WO1995003028A1 (fr) * | 1993-07-23 | 1995-02-02 | Morris Herstein | Composition cosmetique stimulant le renouvellement de la peau, avec effet prolonge d'elimination de l'irritation |
FR2737971A1 (fr) * | 1995-08-25 | 1997-02-28 | Lvmh Rech | Utilisation de la vitamine c ou de ses derives ou analogues pour stimuler la synthese de l'elastine cutanee |
Non-Patent Citations (4)
Title |
---|
CHEMICAL ABSTRACTS, 1 January 1900, Columbus, Ohio, US; abstract no. 117:4934, XP002119696 * |
DATABASE STN EMBASE 1 January 1900 (1900-01-01), XP002119695, Database accession no. 1998376469 * |
PATENT ABSTRACTS OF JAPAN vol. 11, no. 282 (C - 446)<2729> * |
STN, Serveur de Bases de Données, Karlsruhe, DE, Fichier Kosmet, AN=15693 résumé * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE10050274A1 (de) * | 2000-10-09 | 2002-04-18 | Henkel Kgaa | Verfahren zur in vitro Bestimmung der Hautalterung |
Also Published As
Publication number | Publication date |
---|---|
FR2791259B1 (fr) | 2001-08-17 |
UY26063A1 (es) | 2000-10-31 |
TWI225410B (en) | 2004-12-21 |
AR018230A1 (es) | 2001-10-31 |
JP2002541074A (ja) | 2002-12-03 |
AU3172600A (en) | 2000-10-09 |
FR2791259A1 (fr) | 2000-09-29 |
EP1165033A1 (fr) | 2002-01-02 |
CA2364608A1 (fr) | 2000-09-28 |
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