WO2002053125A2 - Produit de soin de la peau contenant un retinoide et systeme renforçateur de l'action du retinoide dans un emballage a compartiment double - Google Patents

Produit de soin de la peau contenant un retinoide et systeme renforçateur de l'action du retinoide dans un emballage a compartiment double Download PDF

Info

Publication number
WO2002053125A2
WO2002053125A2 PCT/EP2001/014769 EP0114769W WO02053125A2 WO 2002053125 A2 WO2002053125 A2 WO 2002053125A2 EP 0114769 W EP0114769 W EP 0114769W WO 02053125 A2 WO02053125 A2 WO 02053125A2
Authority
WO
WIPO (PCT)
Prior art keywords
retinoid
retinol
skin
retinyl
composition
Prior art date
Application number
PCT/EP2001/014769
Other languages
English (en)
Other versions
WO2002053125A3 (fr
Inventor
Stewart Paton Granger
Sreekumar Pillai
Ian Richard Scott
Original Assignee
Unilever Plc
Unilever Nv
Hindustan Lever Limited
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Unilever Plc, Unilever Nv, Hindustan Lever Limited filed Critical Unilever Plc
Priority to AU2002226373A priority Critical patent/AU2002226373A1/en
Publication of WO2002053125A2 publication Critical patent/WO2002053125A2/fr
Publication of WO2002053125A3 publication Critical patent/WO2002053125A3/fr

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
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/07—Retinol compounds, e.g. vitamin A
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34—Alcohols
    • A61K8/342—Alcohols having more than seven atoms in an unbroken chain
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34—Alcohols
    • A61K8/345—Alcohols containing more than one hydroxy group
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/35—Ketones, e.g. benzophenone
    • 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
    • A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
    • 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
    • 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
    • A61K8/671—Vitamin A; Derivatives thereof, e.g. ester of vitamin A acid, ester of retinol, retinol, retinal
    • 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
    • A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
    • A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
    • 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
    • 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
    • A61K2800/88—Two- or multipart kits
    • 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 skin care product containing a retinoid and a retinoid booster in a dual compartment package.
  • Retinoids e.g. retinol and retinyl esters
  • Retinol vitamin A
  • Natural and synthetic vitamin A derivatives have been used extensively in the treatment of a variety of skin disorders and have been used as skin repair or renewal agents.
  • Retinoic acid has been employed to treat a variety of skin conditions, e.g., acne, wrinkles, psoriasis, age spots and discoloration. See, e.g., Vahlquist, A. et al., J. Invest. Dermatol., Vol. 94, Holland D.B.
  • Retinol is particularly unstable in cosmetic formulations because retinol can undergo chemical degradation as a consequence of many factors which include oxidation, thermal instability and UV induced degradation.
  • Retinyl esters are also subject to these instabilities although to a lesser extent than retinol.
  • Retinoid benefits on skin can be enhanced by the coapplication of retinoid booster molecules.
  • many if not all of the retinoid booster molecules also increase the instability of the retinol. Therefore, in order to have an effective skin care composition containing both retinoids and retinoid boosting molecules, it is necessary to protect retinoid formulations containing boosters to a higher degree than is necessary for formulations containing retinoids alone.
  • U.S. Patent No. 5,976,555 assigned to Johnson & Johnson discloses skin care compositions comprising oil in water emulsions containing retinoids, an emulsifier system, and a co-emulsifier .
  • the patent describes the use of a container for storing the composition so that the composition is out of contact with oxygen.
  • the container is described for use for the retinoid composition with an emulsifier system and a co-emulsifier alone and does not protect the retinoid from degradation due to contact with retinoid boosters.
  • the present invention provides a stable skin care product containing:
  • a first composition comprising about 0.001% to about 10% of a retinoid
  • a second composition comprising about 0.0001% to about 50% of at least one retinoid booster
  • a second compartment for storing the second composition, the first and second compartments being joined together.
  • compositions contain, as a preferred ingredient, a retinoid, which is selected from retinyl esters, retinol, retinal and retinoic acid, preferably retinol or retinyl ester.
  • retinol includes the following isomers of retinol: all-trans-retinol, 13-cis- retinol, 11-cis-retinol, 9-cis-retinol, 3, 4-didehydro- retinol, 3, 4-didehydro-13-cis-retinol; 3, 4-didehydro-ll-cis- retinol; 3, 4-didehydro-9-cis-retinol.
  • Preferred isomers are all-trans-retinol, 13-cis-retinol, 3, 4-didehydro-retinol, 9- cis-retinol. Most preferred is all-trans-retinol, due to its wide commercial availability.
  • Retinyl ester is an ester of retinol.
  • the term "retinol" has been defined above.
  • Retinyl esters suitable for use in the present invention are C -C30 esters of retinol, preferably C2-C 2 0 esters, and most preferably C2, C3, and C Q esters because they are more commonly available.
  • retinyl esters include but are not limited to: retinyl palmitate, retinyl formate, retinyl acetate, retinyl propionate, retinyl butyrate, retinyl valerate, retinyl isovalerate, retinyl hexanoate, retinyl heptanoate, retinyl octanoate, retinyl nonanoate, retinyl decanoate, retinyl undecanoate, retinyl laurate, retinyl tridecanoate, retinyl myristate, retinyl pentadecanoate, retinyl heptadecanoate, retinyl stearate, retinyl isostearate, retinyl nonadecanoate, retinyl arachidonate, retinyl behenate, retin
  • the preferred ester for use in the present invention is selected from retinyl palmitate, retinyl acetate and retinyl propionate, because these are the most commercially available and therefore the cheapest.
  • Retinyl linoleate and retinyl oleate are also preferred due to their efficacy.
  • Retinol or retinyl ester is employed in the inventive composition in an amount of about 0.001% to about 10%, preferably in an amount of about 0.01% to about 1%, most preferably in an amount of about 0.01% to about 0.5%.
  • retinoids are enzymatically converted in the skin into retinoic acid according to the mechanism described in Chart 1.
  • Retinyl y Retinol Retinal Cytochrome ester de ydrogenase de ydrogenase P450 hydrolase (B2) (B5)
  • ARAT/LRAT Acyl Coenzyme A (CoA) : Retinol Acyl Transferase/Lecithin: Retinol Acyl Transferase
  • CRABPII Cellular Retinoic Acid Binding Protein II
  • boosters are collectively termed herein as "boosters" and are coded as groups Bl through B5, as can be seen in Chart 1 hereinabove.
  • the boosters alone or in combination with each other, potentiate the action of a retinoid by increasing the amount of retinol available for conversion to retinoic acid and inhibiting the degradation of retinoic acid.
  • the boosters act in conjunction with a retinoid (e.g.
  • compositions include a retinoid in the composition, co-present with a booster, to optimize performance.
  • the present invention includes, in part, a second composition containing about 0.0001% to about 50%, preferably 0.001% to 10%, most preferably from about 0.001% to about 5% by weight of the composition of at least one booster compound, wherein the compound, either alone or at a combined concentration of lOmM inhibit transglutaminase in an in vivo transglutaminase assay to more than 50%, and a cosmetically acceptable vehicle.
  • the boosters included in the inventive compositions are preferably selected from:
  • B1/B2; B1/B3; B1/B4; B1/B5; B2/B3, B2/B4; B2/B5, B3/B4; B3/B5; B4/B5 (c) ternary combinations of boosters selected from the group consisting of
  • the preferred compositions include at least one booster from the different groups (i.e., groups (b) through (e) above) .
  • groups (b) through (e) above any combination of boosters chosen from the different groups may also be employed in the inventive compositions for desired boosting effects.
  • the compounds included in the present invention as boosters are first selected based on the ability of such compounds to pass, at a certain concentration listed in Table A, an in-vitro Microsomal Assay for a specific enzyme as described below under sections 2.1 through 2.7.
  • transglutaminase assay (alone or in combination with another booster) is then subjected to an in vitro transglutaminase assay described below, at an individual or combined concentration of 10 mM. If such combination inhibits transglutaminase to more than 50%, then it is suitable for use in the present invention. If a booster was tested individually, and passes the transglutaminase assay, then it may be combined with another booster or combination that passes the transglutaminase assay.
  • compositions according to the present invention contain combinations of boosters which at an individual concentration of 10 mM inhibit transglutaminase to more than 50%.
  • condition means prevention and treatment of dry skin, acne, photodamaged skin, appearance of wrinkles, age spots, aged skin, increasing stratum corneum flexibility, lightening skin color, controlling sebum excretion and generally increasing the quality of skin.
  • the composition may be used to improve skin desquamation and epidermal differentiation.
  • a booster is a compound which passes an in vitro Microsomal Assay described below in sections 2.1 through 2.7.
  • a compound of the present invention inhibits or enhances at a concentration listed in Table A, an enzyme, to at least a broad % listed in Table A.
  • All-trans-retinol, all-trans-retinoic acid, pal itoyl- CoA, dilauroyl phosphatidyl choline, NAD, and NADPH were purchased from Sigma Chemical Company.
  • Stock solutions of retinoids for the microsomal assays were made up in HPLC grade acetonitrile. All retinoid standard stock solutions for HPLC analysis were prepared in ethanol, stored under atmosphere of N 2 at -70 °C and maintained on ice under amber lighting when out of storage.
  • Other chemicals and the inhibitors were commercially available from cosmetic material suppliers or chemical companies such as Aldrich or International Flavors and Fragrances.
  • Each eyecup was washed with 2x 0.5mL cold buffer (0.1M P04 / ImM DTT / 0.25M sucrose, pH 7 ) by rubbing the darkly pigmented cells with an artist's brush or a rubber policeman.
  • the cell suspension was added to the iridescent membranes and the suspension was stirred for several minutes in a beaker with a Teflon stir bar.
  • the suspension was filtered through a coarse filter (Spectra / Por 925 ⁇ pore size polyethylene mesh) to remove large particles, and the resulting darkly colored suspension was homogenized using a Glas-Col with a motor driven Teflon homogenizer.
  • the cell homogenate was centrifuged for 30 min.
  • the resulting homogenate was successively centrifuged for 30 min. at 10,000g, 30 min. at 20,000g, and 15 min. at 30,000g, and the resulting supernatant was ultracentrifuged for 80 min. at 105,000g.
  • the pellet was sonicated in ⁇ 5mL of 0.1M P04 / 0. ImM EDTA / 5mM MgC12, pH 7.4 buffer as described above and stored as- aliquots at - 70°C Protein concentrations were determined as described above.
  • the incubation solution contains 40 ⁇ M acyl donor, lOO ⁇ M or less inhibitor, lO ⁇ M retinol, approximately 30 ⁇ g/mL microsomal protein, and nearly 0. IM P04, pH 7 / 5mM DTT / 2mg/mL BSA. All steps subsequent to the addition of retinol were done in the dark or under amber lights.
  • CRABPII a. System of expression The gene CRABPII was cloned in pET 29a-c(+) plasmid (Novagen) . The cloned gene was under control of strong bacteriophage T7 transcription and translation signals. The source of T7 polymerase was provided by the host cell E.coli BLR(DE3)pLysS (Novagen). The latter has a chromosomal copy of T7 polymerase under lac ⁇ V5 control, induced by the presence of IPTG. The plasmid was transferred into E. coli BLR(DE3)pLysS cells by transformation according to the manufacturer protocol (Novagen) .
  • the frozen pellet was thawed at RT and resuspended in 1- 2 pellet volumes of freshly prepared lysis buffer (50 mM
  • Tris-Hcl pH 8, 10%(w/v) sucrose, 1 mM EDTA, 0.05% (w/v) sodium azide, 0.5 mM DTT, 10 mM MnC12, 2.5 mM phenylmethylsulfonyl fluoride, 2.5 mM benzamidine, 6 ⁇ g/mL DNase) .
  • the lysate was incubated for 30 min at room temperature. Further lysis was accomplished by sonication (six 30-sec bursts at 10,000 psi alternated with five 30-sec delay on ice) .
  • the insoluble fraction of the lysate was removed by centrifugation at 15000 rpm 1 hour at 4°C and the supernatant is stored at -20°C
  • step a The supernatant from step a. was loaded onto a 2.5x100 cm column of sephacryl S-300 (Pharmacia) at room temperature.
  • the elution buffer was 20 mM Tris-HCl, pH 8, 0.5mM DTT, 0.05% sodium azide (buffer A) .
  • the flow rate was 2mL/min. Collected 2-mL fractions were checked for ultraviolet absorbance at 280 nm. The fractions representing the peaks were examined by
  • CRABPII was stored at 4°C before freeze-drying using a Micromodulyo 1.5K with vial platform attachment (Edwards High Vacuum International) . The desiccated samples were stored at room temperature until their use in the binding assay. d. Detection of the presence of CRABPII
  • CRABPII denaturing SDS-polyacrylamide gel electrophoresis (SDS- PAGE) analysis on a 7-15% polyacrylamide gel (Biorad) .
  • 10 ⁇ L samples were mixed with 10 ⁇ L of 2X loading buffer (100 mM Tris-HCl pH6.8, 4% SDS, 0.2% BPB, 20% glycerol, ImM DTT) and denatured by heating (2 min at 80°C) .
  • the samples were loaded onto the gel that was immersed in a IX Tris-glycine buffer (Biorad) and a constant current (25 mA) was applied for 1 hour at room temperature. After Coomassie blue staining, the protein was identified according to its molecular weight as determinated with the Benchmark prestained protein ladder (Gibco BRL) .
  • a western blot was used to confirm the presence of CRABPII.
  • the proteins separated on the SDS-PAGE were transferred on an Immobilon-P transfer membrane (Millipore) using a Biorad cassette. The transfer occurred in IX Tris- glycine buffer (Biorad) + 10% methanol . An electrical current (60 mA) was applied for 3 hours to allow the protein to migrate through the membrane. Afterwards, the membrane was blocked with 5% dry milk in IX TBS for one hour at room temperature and probed with primary antibodies to CRABPII
  • the final incubation solution contains 2.4mM NADPH, lOO ⁇ M or less inhibitor, lO ⁇ M retinoic acid, approximately 4mg/mL rat liver microsomal protein and nearly 0. IM P04 /O.lmM EDTA / 5mM MgC12.
  • Samples for retinoid quantitation by HPLC were prepared by dissolving the residue in each vial with lOO ⁇ L of methanol. The solution was transferred to a 150 ⁇ L glass conical tube within a lmL shell vial, capped tightly, and placed inside a Waters 715 Autosampler. Aliquots of 60 ⁇ L were injected immediately and analyzed for retinoid content.
  • the chromatography instrumentation consisted of a Waters 600 gradient controller / pump, a Waters 996 Photodiode Array detector and a Waters 474 Scanning Fluorescence detector. Two HPLC protocols were used for retinoid analysis.
  • ARAT and LRAT assay the separation of retinol and retinol esters was performed with a Waters 3.9x300mm C18 Novapak reverse-phase analytical column and Waters Sentry NovaPak C18 guard column with an 80:20 (v/v) methanol / THF isocratic mobile phase adjusted to a flow rate of lmL/min. for 10 min. The eluate was monitored for absorbance at 325nm and fluorescence at 325ex/480em.
  • boosters suitable for further testing in the transglutaminase assay include but are not limited to the boosters listed in Tables Bl through B5 below.
  • Inhibition TG (IC 50) Inhibition Inhibition Inhibition Inhibition Inhibition Inhibition Overall ARAT (10pm) ARAT(lOO ⁇ m) LRAT LRAT TG (lO ⁇ m) (lOO ⁇ m) (- ROH/RE)
  • Phospholipid Phosphatidyl Choline 21% increase Phospholipid Sphingomyelin 26% increase
  • the boosters or combinations thereof inhibit transglutaminase (hereinafter "Tgase”) in a transglutaminase assay described below to at least 50% at a concentration of lOmM.
  • Tegase transglutaminase
  • a 15nm thick layer of protein known as the cornified envelope (CE) is formed on the inner surface of the cell periphery.
  • the CE is composed of numerous distinct proteins which have been cross-linked together by the formation of N — (Y-glutamyl) lysine isodipeptide bonds catalyzed by the action of at least two different transglutaminases (TGases) expressed in the epidermis.
  • TGases transglutaminases
  • TGase I is a useful marker of epidermal keratinocyte differentiation with high TGase I levels indicating a more differentiated state.
  • Keratinocytes (cultured as described above) were plated in 96 well plates at a density of 4,000-5,000 cells per well in 200 ⁇ l media. After incubation for two to three days, or until cells are ⁇ 50% confluent, the media was changed to media containing test compounds (five replicates per test) . The cells were cultured for a further 96 hours after which time the media was aspirated and the plates stored at -70°C. Plates were removed from the freezer, and the cells were washed twice with 200 ⁇ l of lx PBS. The cells were incubated for one hour at room temperature (R/T) with TBS/5% BSA (wash buffer, bovine serum albumin) .
  • R/T room temperature
  • BSA wash buffer, bovine serum albumin
  • TGase primary antibody 50 ⁇ l of monoclonal anti-Tgase I Ab B.C. diluted 1:2000 in wash buffer. The primary antibody was incubated for 2 hours at 37°C and then rinsed 6x with wash buffer. Cells were then incubated with 50 ⁇ l of secondary antibody (Fab fragment, peroxidase conjugated anti-mouse IgG obtaining from Amersham) diluted 1:4,000 in wash buffer for two hours at 37°C, then rinsed three times with wash buffer. Following the rinse with washing buffer, the cells were rinsed 3x with PBS.
  • secondary antibody Fab fragment, peroxidase conjugated anti-mouse IgG obtaining from Amersham
  • the cells were incubated with lOO ⁇ l substrate solution (4 mg o- phenylenediamine and 3.3 ⁇ l 30% H 2 O 2 in 10ml 0.1M citrate buffer pH 5.0) for exactly five minutes, R/T, in darkness (under aluminum foil) .
  • the reaction was stopped by the addition of 50 ⁇ l 4N H 2 SO 4 .
  • the absorbance of samples was read at 492nm in a 96 well plate UV spectrophotometer. Out of the five replicates, four were treated with both antibodies, the fifth one was used as a Tgase background control. TGase levels were determined and expressed as percentage control.
  • Transglutaminase levels were determined and expressed in the Tables Bl through B5 above either as:
  • compositions which include retinoids are generally unstable and may undergo chemical degradation. Moreover, it has been surprisingly found that boosters, although beneficial for enhancing the retinoid benefits, also contribute to the chemical instability of retinoids. The booster induced retinol destabilization dramatically reduces the overall efficacy of the boosted retinoid composition when both ingredients are contained in a single formula. Therefore, in order to protect against retinoid breakdown while still providing the beneficial effects of retinoid boosters, the present invention provides a dual compartment package that contains a first composition containing retinoids in a first compartment and a second composition containing at least one retinoid booster in a second compartment. The first composition provides a first benefit to the skin while the second composition works to boost or enhance the effect of the first benefit.
  • the dual compartment package may be designed in various ways known to persons of ordinary skill in the art as long as the purpose of providing the first and second compositions in two separate containers is achieved.
  • the dual compartment package is in the form of two jars or bottles adjoiningly attached.
  • the dual compartment package is in the form of a single bottle/jar with a division separating an interior of the bottle/jar into a first and second compartment.
  • Other embodiments are contemplated as being within the scope of the present invention as long as the compositions are retained separately.
  • the product according to the present invention also comprises a cosmetically acceptable vehicle to act as a dilutant, dispersant, or carrier for the active components in the either or both the first and second compositions, so as to facilitate their distribution when the composition is applied to the skin.
  • Vehicles other than or in addition to water can include liquid or solid emollients, solvents, humectants, thickeners and powders.
  • An especially preferred nonaqueous carrier is a polydimethyl siloxane and/or a polydimethyl phenyl siloxane.
  • Silicones of this invention may be those with viscosities ranging anywhere from about 10 to 10,000,000 centistokes at 25°C. Especially desirable are mixtures of low and high viscosity silicones. These silicones are available from the General Electric Company under trademarks Vicasil, SE and SF and from the Dow Corning Company under the 200 and 550 Series. Amounts of silicone which can be utilized in the compositions of this invention range anywhere from 5 to 95%, preferably from 25 to 90% by weight of the composition.
  • an oil or oily material may be present, together with an emulsifier to provide either a water-in-oil emulsion or an oil-in-water emulsion, depending largely on the average hydrophilic- lipophilic balance (HLB) of the emulsifier employed.
  • HLB hydrophilic- lipophilic balance
  • Actives are defined as skin or hair benefit agents other than emollients and other than ingredients that merely improve the physical characteristics of the composition. Although not limited to this category, general examples include sunscreens, skin lightening agents, tanning agents.
  • Sunscreens include those materials commonly employed to block ultraviolet light.
  • Illustrative compounds are the derivatives of PABA, cinnamate and salicylate.
  • cinnamate and salicylate For example, ' octyl methoxycinnamate and 2-hydroxy-4-methoxy benzophenone
  • Octyl methoxycinnamate and 2-hydroxy-4-methoxy benzophenone are commercially available under the trademarks Parsol MCX and
  • the exact amount of sunscreen employed in the emulsions can vary depending upon the degree of protection desired from the sun's UV radiation.
  • EFAs essential fatty acids
  • keratinocytes EFA deficiency makes cells hyperproliferative. Supplementation of EFA corrects this. EFAs also enhance lipid biosynthesis of epidermis and provide lipids for the barrier formation of the epidermis .
  • the essential fatty acids are preferably chosen from linoleic acid, Y-linolenic acid, homo-Y-linolenic acid, columbinic acid, eicosa- (n-6, 9, 13) -trienoic acid, arachidonic acid, Y- linolenic acid, timnodonic acid, hexaenoic acid and mixtures thereof.
  • Emollients are often incorporated into cosmetic compositions of the present invention. Levels of such emollients may range from about 0.5% to about 50%, preferably between about 5% and 30% by weight of the total composition. Emollients may be classified under such general chemical categories as esters, fatty acids and alcohols, polyols and hydrocarbons .
  • Esters may be mono- or di-esters.
  • Acceptable examples of fatty di-esters include dibutyl adipate, diethyl sebacate, diisopropyl dimerate, and dioctyl succinate.
  • Acceptable branched chain fatty esters include 2-ethyl-hexyl myristate, isopropyl stearate and isostearyl palmitate.
  • Acceptable tribasic acid esters include triisopropyl trilinoleate and trilauryl citrate.
  • Acceptable straight chain fatty esters include lauryl palmitate, myristyl lactate, oleyl eurcate and stearyl oleate.
  • Preferred esters include coco- caprylate/caprate (a blend of coco-caprylate and coco- caprate) , propylene glycol myristyl ether acetate, diisopropyl adipate and cetyl octanoate.
  • Suitable fatty alcohols and acids include those compounds having from 10 to 20 carbon atoms. Especially preferred are such compounds such as cetyl, myristyl, palmitic and stearyl alcohols and acids.
  • polyols which may serve as emollients are linear and branched chain alkyl polyhydroxyl compounds.
  • propylene glycol, sorbitol and glycerin are preferred.
  • polymeric polyols such as polypropylene glycol and polyethylene glycol.
  • Butylene and propylene glycol are also especially preferred as penetration enhancers .
  • hydrocarbons which may serve as emollients are those having hydrocarbon chains anywhere from 12 to 30 carbon atoms. Specific examples include mineral oil, petroleum jelly, squalene and isoparaffins .
  • thickeners are also categories of functional ingredients within the cosmetic compositions of the present invention.
  • a thickener will usually be present in amounts anywhere from 0.1 to 20% by weight, preferably about 0.5% to about 10% by weight of the composition.
  • Exemplary thickeners are cross-linked polyacrylate materials available under the trademark Carbopol from the B.F. Goodrich Company. Gums may be employed such as xanthan, carrageenan, gelatin, karaya, pectin and locust beans gum. Under certain circumstances the thickening function may be accomplished by a material also serving as a silicone or emollient. For instance, silicone gums in excess of 10 centistokes and esters such as glycerol stearate have dual functionality.
  • Powders may be incorporated into one or both of the first and second cosmetic compositions of the cosmetic product of the present invention. These powders include chalk, talc, Fullers earth, kaolin, starch, smectite clays, chemically modified magnesium aluminum silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, aluminum starch octenyl succinate and mixtures thereof. Other adjunct minor components may also be incorporated into one or both of the first and second compositions of the cosmetic product of the present invention. These ingredients may include coloring agents, opacifiers and perfumes. Amounts of these materials may range anywhere from 0.001% up to 20% by weight of the composition.
  • compositions of the cosmetic product of the present invention are intended primarily as a product for topical application to human skin, especially as an agent for conditioning and smoothening the skin, and preventing or reducing the appearance of wrinkled or aged skin.
  • a small quantity of the first composition for example from 1 to 5ml, is applied to exposed areas of the skin, from a suitable container or applicator and, if necessary, it is then spread over and/or rubbed into the skin using the hand or fingers or a suitable device.
  • a small quantity of the second composition for example from 1 to 5 ml, is applied to exposed areas of the skin, from a suitable container or applicator and, if necessary, it is also spread over and/or rubbed into the skin using the hand or fingers or a suitable device. Therefore, depending upon the intensity of treatment benefits desired, the first and second compositions may be used alone, simultaneously, or in consecutive order.
  • the topical skin treatment composition of the invention can be formulated as a lotion, a fluid cream, a cream or a gel.
  • Retinol (50% in tween 80) was dissolved in approximately 50% aqueous ethanol to provide a solution giving an OD at 360nm of approximately 0.6 when measured in a 200 ⁇ l volume in a 96 well plate using a standard 96 well spectrophotometer .
  • Booster molecules were added at approximately 0.1% concentration and the OD 360 measured as above immediately and after 60 hours at room temperature in the dark. A correction was applied to the OD after 60 hours (divide by 0.85) to account for increased concentration of the retinol due to evaporation of solvent from the plate.
  • Boosters tested caused marked increases in the instability of the retinol. This will make it necessary to use formulation/packaging options providing considerably better stability to the retinol when boosters are used compared to those needed for retinol alone.
  • concentration leaves a window of 40-100% for further transglutaminase inhibition for detecting synergy of the two compounds under examination.
  • a more challenging concentration criteria would be selecting concentrations of compounds which alone showed no boosted retinol inhibition of transglutaminase. In this study, however, even more challenging criteria were chosen. Concentrations of compounds that were 10 fold and 100 fold lower than the minimally effective transglutaminase inhibiting concentration were selected. Identification of synergistic combinations using such very low concentrations would mean that the most effective synergistic combinations were identified.
  • the data in the following table represents the concentrations of compound that are 2 logs lower than the minimally inhibitory compound concentration. These were the concentrations used in the B1/B5 combination studies.
  • the efficacy of the B1/B5 combinations splits into two classes - particularly effective combinations (bolded in the above table, i.e., the first 14 combinations) and barely effective combinations (not bolded, i.e., the latter 6 combinations) . It was unexpected that certain B1/B5 combinations performed better than other combinations. Those combinations which were barely effective were (i) fatty acid amides + azoles (ii) hydroxy fatty acid amides + azoles and (iii) naringenin/quercetin + azoles.
  • Bl boosters combined with B5 boosters from the following classes: fatty hydroxyethyl imidazoline surfactants, cyclic aliphatic unsaturated compounds, polycyclic triterpenes, n-substituted fatty acid amides.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Birds (AREA)
  • Emergency Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Dermatology (AREA)
  • Cosmetics (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

L'invention concerne un produit stable de soin pour la peau contenant une première composition à base d'environ 0,001 à 10 % de rétinoïde, une seconde composition comprenant environ 0,0001 à 50 % d'au moins un renforçateur de l'action du rétinoïde, un premier compartiment pour stocker la première composition et un second compartiment pour stocker la seconde composition, le premier et le second compartiment étant réunis.
PCT/EP2001/014769 2000-12-28 2001-12-13 Produit de soin de la peau contenant un retinoide et systeme renforçateur de l'action du retinoide dans un emballage a compartiment double WO2002053125A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002226373A AU2002226373A1 (en) 2000-12-28 2001-12-13 Skin care product containing a retinoid and a retinoid booster system in a dual compartment package

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US25846000P 2000-12-28 2000-12-28
US60/258,460 2000-12-28

Publications (2)

Publication Number Publication Date
WO2002053125A2 true WO2002053125A2 (fr) 2002-07-11
WO2002053125A3 WO2002053125A3 (fr) 2002-10-03

Family

ID=22980635

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2001/014769 WO2002053125A2 (fr) 2000-12-28 2001-12-13 Produit de soin de la peau contenant un retinoide et systeme renforçateur de l'action du retinoide dans un emballage a compartiment double

Country Status (2)

Country Link
AU (1) AU2002226373A1 (fr)
WO (1) WO2002053125A2 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009158499A2 (fr) * 2008-06-25 2009-12-30 University Of North Texas Health Science Center At Fort Worth Prévention de la croissance bactérienne et de la formation de biofilm par des ligands qui agissent sur les systèmes cannabinoïdergiques
US20120283235A1 (en) * 2009-10-20 2012-11-08 Discovery Partners Llc Dermatologic and Cosmetic Compositions
EP3265053A4 (fr) * 2015-03-05 2018-08-08 Avon Products, Inc. Procédés de traitement de la peau
US10076479B1 (en) 2018-05-08 2018-09-18 Avon Products, Inc. Methods for treating skin

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0646371A1 (fr) * 1993-10-05 1995-04-05 L'oreal Composition cosmétique ou pharmaceutique, anhydre et à base de rétinol
WO1996037420A1 (fr) * 1995-05-26 1996-11-28 Unilever Plc Cure de soins cutanes
WO1997031620A2 (fr) * 1996-03-01 1997-09-04 Johnson & Johnson Consumer Products, Inc. Compositions a usage local contenant une emulsion aqueuse et un retinoide
WO1998013018A1 (fr) * 1996-09-27 1998-04-02 Unilever Plc Composition pour le soin de la peau contenant une amide, et du retinol ou un ester de retinyle
WO1998013017A1 (fr) * 1996-09-27 1998-04-02 Unilever Plc Compositions pour le soin de la peau contenant une amide et un retinoide
WO1998013020A1 (fr) * 1996-09-27 1998-04-02 Unilever Plc Compositions pour le soin de la peau contenant des combinaisons de composes permettant de reproduire l'effet sur la peau de l'acide retinoique
EP0995433A1 (fr) * 1998-10-13 2000-04-26 JOHNSON & JOHNSON CONSUMER COMPANIES, INC. Compositions de nettoyage stabilisées contenant du rétinol

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5976555A (en) * 1994-09-07 1999-11-02 Johnson & Johnson Consumer Products, Inc. Topical oil-in-water emulsions containing retinoids

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0646371A1 (fr) * 1993-10-05 1995-04-05 L'oreal Composition cosmétique ou pharmaceutique, anhydre et à base de rétinol
WO1996037420A1 (fr) * 1995-05-26 1996-11-28 Unilever Plc Cure de soins cutanes
WO1997031620A2 (fr) * 1996-03-01 1997-09-04 Johnson & Johnson Consumer Products, Inc. Compositions a usage local contenant une emulsion aqueuse et un retinoide
WO1998013018A1 (fr) * 1996-09-27 1998-04-02 Unilever Plc Composition pour le soin de la peau contenant une amide, et du retinol ou un ester de retinyle
WO1998013017A1 (fr) * 1996-09-27 1998-04-02 Unilever Plc Compositions pour le soin de la peau contenant une amide et un retinoide
WO1998013020A1 (fr) * 1996-09-27 1998-04-02 Unilever Plc Compositions pour le soin de la peau contenant des combinaisons de composes permettant de reproduire l'effet sur la peau de l'acide retinoique
EP0995433A1 (fr) * 1998-10-13 2000-04-26 JOHNSON & JOHNSON CONSUMER COMPANIES, INC. Compositions de nettoyage stabilisées contenant du rétinol

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009158499A2 (fr) * 2008-06-25 2009-12-30 University Of North Texas Health Science Center At Fort Worth Prévention de la croissance bactérienne et de la formation de biofilm par des ligands qui agissent sur les systèmes cannabinoïdergiques
WO2009158499A3 (fr) * 2008-06-25 2011-03-24 University Of North Texas Health Science Center At Fort Worth Prévention de la croissance bactérienne et de la formation de biofilm par des ligands qui agissent sur les systèmes cannabinoïdergiques
US20120283235A1 (en) * 2009-10-20 2012-11-08 Discovery Partners Llc Dermatologic and Cosmetic Compositions
EP3265053A4 (fr) * 2015-03-05 2018-08-08 Avon Products, Inc. Procédés de traitement de la peau
US10123954B1 (en) 2015-03-05 2018-11-13 Avon Products, Inc. Methods for treating skin
US10328012B2 (en) 2015-03-05 2019-06-25 Avon Products, Inc. Methods for treating skin
EP3578163A1 (fr) * 2015-03-05 2019-12-11 Avon Products, Inc. Procédés de traitement de la peau
US10828243B2 (en) 2015-03-05 2020-11-10 Avon Products, Inc. Methods for treating skin
US10076479B1 (en) 2018-05-08 2018-09-18 Avon Products, Inc. Methods for treating skin

Also Published As

Publication number Publication date
WO2002053125A3 (fr) 2002-10-03
AU2002226373A1 (en) 2002-07-16

Similar Documents

Publication Publication Date Title
EP1333800B1 (fr) Compositions pour le soin de la peau renfermant des composes qui reproduisent l'effet de l'acide retinoique sur la peau
EP1349536B1 (fr) Compositions stables pour les soins de la peau contenant un retinoide et systeme renfor ant l'action du retinoide
AU2002229642B2 (en) Skin care product containing retinoids, retinoid booster and phytoestrogens in a dual compartment package
AU2002229642A1 (en) Skin care product containing retinoids, retinoid booster and phytoestrogens in a dual compartment package
AU2002216090A1 (en) Stable skin care compositions containing a retinoid and a retinoid booster system
AU2002235762B2 (en) Stable skin conditioning compositions containing retinoid boosters
AU2002235762A1 (en) Stable skin conditioning compositions containing retinoid boosters
EP1345586B1 (fr) Produit de soin cutane stable contenant un retinoide et un systeme activateur de retinoide dans un emballage a deux compartiments
AU2002234562A1 (en) Stable skin care product containing a retinoid and a retinoid booster system in a dual compartment package
WO2002053125A2 (fr) Produit de soin de la peau contenant un retinoide et systeme renforçateur de l'action du retinoide dans un emballage a compartiment double

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PH PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

AK Designated states

Kind code of ref document: A3

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PH PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP