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WO2003006009A1 - Compositions permettant de reduire la cellulite ou de prevenir son apparition chez les mammiferes - Google Patents

Compositions permettant de reduire la cellulite ou de prevenir son apparition chez les mammiferes Download PDF

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Publication number
WO2003006009A1
WO2003006009A1 PCT/US2002/011079 US0211079W WO03006009A1 WO 2003006009 A1 WO2003006009 A1 WO 2003006009A1 US 0211079 W US0211079 W US 0211079W WO 03006009 A1 WO03006009 A1 WO 03006009A1
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Prior art keywords
linoleic acid
conjugated linoleic
trans
cis
composition
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PCT/US2002/011079
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English (en)
Inventor
Anindita Sen
Peter Pieraccini
Yolanda Renee Lea-Currie
Dawn Marie Franklin
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Zen-Bio, Inc.
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Publication of WO2003006009A1 publication Critical patent/WO2003006009A1/fr

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Classifications

    • 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
    • A61Q19/06—Preparations for care of the skin for countering cellulitis
    • 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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • 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
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/361—Carboxylic acids having more than seven carbon atoms in an unbroken chain; Salts or anhydrides 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
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions

Definitions

  • the present invention relates to compositions, comprising conjugated linoleic acid and an imidazole antifungal agent, and methods that are useful for the treatment and prevention of cellulite in mammalian skin and for promoting body slimming by reducing localized fatty excesses. More specifically, the invention provides skin care compositions that can be used to inhibit lipogenesis and/or stimulate lipolysis in human adipocytes.
  • Cellulite is a term applied to a skin condition associated with the lumps, bumps and dimples that appear on the thighs of many women.
  • Cellulite primarily afflicts the thighs and buttocks but may also be present on the stomach and upper arms. This condition is frequently described as “orange peel skin”, “mattress phenomena” or the “cottage cheese effect.”
  • Cellulite afflictions are a stubborn problem causing emotional and psychological distress to many women.
  • the etiology of cellulite is poorly understood, the main etio logical factor appears to be local accumulation of fat in a regional compartment.
  • Adipocytes are the principle cells implicated in fat storage by adipose tissue. It has been proposed that the anatomical structure of subcutaneous adipose tissue is the major cause of cellulite. The histological studies of subcutaneous tissues from men and women suggest that the fat lobules are larger and more vertical in women than men. As a result, these larger, less restricted lobules can express outward against the dermis causing the bumps and dimples characteristic of cellulite. The femoral subcutaneous fat deposits in women also tend to be more lipogenic and less lipolytic than abdominal subcutaneous or visceral fat due to the difference in the distribution of ⁇ and ⁇ adrenergic receptors on adipocytes in these different regions. Increased lipolysis or fat reduction of these selected subcutaneous adipose sites may lead to a reduction or the prevention of cellulite.
  • the most commonly known and used is that which consists in inhibiting the phosphodiesterase in order to prevent or at least limit the rate of degradation of cyclic AMP.
  • the phosphodiesterase destroys cyclic AMP by transforming it into 5' AMP so that it cannot function as a lipolysis activator.
  • Topical application for the treatment of cellulite of agents capable of distributing or reducing local fat accumulation by lipolytic action thereby improving the aesthetic appearance of the skin has been used.
  • the common agents for treatment of cellulite as slimming agents are xanthine analogs such as caffeine or theophyUine. These agents block the antilipolytic action of adenosine, a potent endogenous inhibitor of lipolysis.
  • Xanthine based adenosine antagonists such as caffeine or theophyUine are also known to be effective phosphodiesterase inhibitors.
  • thermo slimming cosmetic composition containing an oil-soluble plant extract having slimming action.
  • oil-soluble plant extracts are vegetable extracts including, principally, those of climbing ivy (Hedera helix), arnica (Arnica montana), rosemary (Rosmarinus officinalis N), marigold (Calendula officinalis), sage (Salvia officinalis N), ginseng (Panax ginseng), St.
  • Conjugated linoleic acid is a collective acronym connoting a class of positional and geometric conjugated dienoic isomers of the long chain (Cl 8) fatty acid octadecadienoic acid.
  • CLA refers to the cis and trans forms of 9,11- 10,12- and 11,13 -octadecadienoic acid. It is to be understood that as used herein the term CLA includes CLA the free fatty acid form as well as derivatives of CLA.
  • Derivatives include those derived from substitution of the carboxyl group of the acid, such as esters (e.g., triglyceride esters, monoglyceride esters, diglyceride esters, phosphoesters), amides, salts (e.g., alkali metal, alkali earth metal, and ammonium salts), as well as forms derived from substitution of the C18 carbon chain, such as hydroxy and/or ⁇ hydroxy derivatives.
  • esters e.g., triglyceride esters, monoglyceride esters, diglyceride esters, phosphoesters
  • amides amides
  • salts e.g., alkali metal, alkali earth metal, and ammonium salts
  • forms derived from substitution of the C18 carbon chain such as hydroxy and/or ⁇ hydroxy derivatives.
  • CLA is known to be anticarcinogenic and antitumorogenic. It was proposed that the anticarcinogenic effect of CLA may be attributed to its antioxidative properties Chin et al, (1993) J. Agric. Food Chem. 528: 262-271 and the antitumorogenic effects may be mediated by the inhibition of prostaglandin E2 synthesis Liu and Belury, Cancer Lett. 127: 15-22 (1998). Dietary CLA supplementation has been reported to regulate body fat and accumulation and retention in mice rats and chicks Belury et al., ( ⁇ 996) Nutr. Cancer 26: 149- 157;
  • compositions and methods for treating and/or preventing cellulite, and for promoting body slimming by administering a safe and effective amount of a skin care composition are provided.
  • the disclosed composition comprises conjugated linoleic acid (CLA), and an imidazole or a triazole antifungal agent in combination with a pharmaceutically acceptable carrier. More particularly, the composition comprises effective amounts of either a mixture of conjugated linoleic acid isomers, or a single isomer, such as, but not limited to, a 9, 11 isomer, a 10, 12 isomer or an 11, 13 isomer of CLA.
  • a 10-trans, 12-cis (lOt, or a 12c) 9-cis, W-trans (9c, l it) isomer of CLA can be used in combination with an antifungal imidazole or triazole agent, such as, but not limited to, ketoconazole (imidazole) or terconazole (triazole) and an acceptable carrier.
  • an antifungal imidazole or triazole agent such as, but not limited to, ketoconazole (imidazole) or terconazole (triazole) and an acceptable carrier.
  • the topical administration of the disclosed skin care compositions is predicted to promote the reduction of adipose tissue accumulation and to prevent de novo lipogenesis.
  • administration of the compositions is intended to improve dermal appearance by preventing the accumulation of subcutaneous adipose tissues, reducing adipose tissue mass, and decreasing the occurrence and severity of cellulite in mammalian skin.
  • the present invention further relates to a skin care composition
  • a skin care composition comprising from about 0.01% to about 40%, by weight, conjugated linoleic acid and about 0.01% to about 40% by weight ketoconazole in a package for said skin care composition.
  • the skin composition of the invention compare from about .025% - 10% conjugated linoleic acid and imidazole antifungal agent. Unless otherwise indicated, all percentages and ratios used herein are by weight of the total composition. All weight percentages, unless otherwise indicated, are on an actives weight basis. All measurements made are at approximately 25° C, unless otherwise designated.
  • the invention provides compositions suitable for use in the cellulite treatment/preventive use disclosed herein.
  • Suitable skin care compositions for use in the methods described herein include compositions generally comprising at least one isomer of conjugated linoleic acid (CLA) in combination with an imidazole or triazole antifungal agent, (referred to herein as an AZOLE agent) and a pharmaceutically acceptable carrier.
  • CLA conjugated linoleic acid
  • AZOLE agent imidazole or triazole antifungal agent
  • suitable compositions will comprise CLA and AZOLE at concentrations ranging from about 0.01% to about 40% by weight.
  • compositions contemplated by the invention may also include an additional agent selected from the group consisting of phosphodiestrase inhibitors, oleosoluble vegetable extracts, herbal extracts, botanical extracts, hydroxy acids, a desquamatory agent, a sunscreen agent, an anti-oxidant, a retinoid or a mixture of any of the above-listed agents.
  • an additional agent selected from the group consisting of phosphodiestrase inhibitors, oleosoluble vegetable extracts, herbal extracts, botanical extracts, hydroxy acids, a desquamatory agent, a sunscreen agent, an anti-oxidant, a retinoid or a mixture of any of the above-listed agents.
  • the invention also disclose the use of a composition comprising at least one conjugated linoleic acid isomer, and an AZOLE (e.g., an imidazole or triazole) antifungal agent and a pharmaceutically acceptable carrier to stimulate lipolysis in adipoctes localized in a subcutaneous region of mammalian skin that is characterized by adipose tissue accumulation.
  • AZOLE e.g., an imidazole or triazole
  • This embodiment of the invention can be practiced for the purpose of treating or preventing the occurrence of cellulite in the skin region selected for treatment.
  • a composition of the invention formulated for topical application to the skin could be applied to the hip and/or thigh region of a female human patient's leg.
  • the invention discloses the use of the compositions described herein to inhibit de novo lipogenesis in adipocytes localized , in a subcutaneous region of mammalian skin that is characterized by adipose tissue accumulation, and which is therefore, likely to become a site where cellulite is likely to appear.
  • This embodiment of the invention can also be utilized to promote body slimming by reducing localized fatty excesses.
  • the invention disclosed herein also discloses the use of conjugated linoleic acid and an antifungal imidazole agent to manufacture topical skin care compositions suitable for use in anyone of the methods disclosed herein.
  • the skin care compositions of the invention comprise from- about 0.010% to about 40% of the active agents (i.e., conjugated linoleic acid and antifungal imidazole).
  • a skin care composition of the invention may comprise about 0.01%, 0.015%, 0.02%, 0.025%, 0.030%, 0.050%,
  • compositions of the invention may be provided with information about and/or instructions on the use of the combination to prevent and/or treat cellulite.
  • safe and effective amount means an amount of a compound or composition sufficient to significantly induce a positive benefit, preferably a positive skin appearance or feel benefit, including independently the benefits disclosed herein, but low enough to avoid serious side effects, i.e., to provide a reasonable benefit to risk ratio, within the scope of sound judgment of the skilled artisan.
  • Figures 1A and IB provide graphic representations of the effects of conjugated linoleic acid (CLA) and ketoconazole (KETO) either alone or in various combinations on lipolysis of human adipocytes isolated from two different human patients.
  • CLA conjugated linoleic acid
  • KETO ketoconazole
  • the bar graphs provided in Figures 1A and IB indicate the amount of glycerol released from adipocytes in response to stimulation with: CLA alone, l ⁇ M CLA (column 1), 10 ⁇ M CLA (column 2), 100 ⁇ M CLA (column 3); KETO alone 1 ⁇ M KETO (column 4), 10 ⁇ M KETO (column 5), 100 ⁇ M KETO (column 6); or combinations comprising CLA and KETO at various concentrations 10 ⁇ M CLA and 10 ⁇ M KETO (column 7), 100 ⁇ M CLA and 10 ⁇ M KETO (column 8), 10 ⁇ M CLA and 100 ⁇ M KETO (column 9), 100 ⁇ M CLA and 100 ⁇ M KETO (column 10), 100 ⁇ M CLA and 0.1 ⁇ M KETO
  • Figure 2 is a graphic representation of the effects of conjugated linoleic acid (i.e., 10- trans, ⁇ 2-cis conjugated linoleic acid (CLA)) the AZOLE ketoconazole (KETO)), and combinations comprising CLA and KETO on lipolysis of human adipocytes isolated from two alternative sources of adipose tissue (dark bars indicate adipocytes obtained from adipose tissue obtained from a thigh/hip region; light bars indicate adipocytes obtained from abdominal tissue). The concentration of each agent is indicated in the figure legend. All concentrations are in ⁇ M.
  • L011901H adipocytes were obtained from subcutaneous adipose tissue obtained from a patient undergoing thigh/hip liposuction.
  • L011701 adipocytes were obtained from subcutaneous adipose tissue obtained from a patient undergoing abdominal liposuction. The data is displayed as fold increase in glycerol release relative to the concentration of glycerol released by adipocytes cultured in the presence of vehicle only.
  • Figure 3 is a graphic representation of the effects of linoleic acid, a 9-cis, 1 l-trans CLA isomer, a IQ-trans, 12-cis CLA isomer on de no o lipogenesis in cultured human adipocytes. The data is displayed as percentage of 14 C-glucose incorporation into total cellular lipids based on vehicle only controls.
  • Figure 4 provides a graphic representation of the effects of a combination of CLA and KETO on lipolysis in human adipocytes obtained from subcutaneous human adipose tissue (dark bars indicate adipocytes obtained from adipose tissue obtained from a thigh/hip region; light bars indicate adipocytes obtained from abdominal tissue).
  • the data is displayed as fold increase in glycerol release relative to the amount of glycerol that is released in response to stimulation with a negative control sample.
  • CLA designates conjugated linoleic acid
  • KETO designates ketoconazole
  • D3MX designates Isobutylmethylxanthine. All of the values defining concentrations are in ⁇ M.
  • compositions and methods for controlling or reducing localized fatty excesses or cellulite are provided.
  • the compositions comprise conjugate linoleic acid (CLA), and an antifungal AZOLE in combination with a pharmaceutically acceptable carrier.
  • Conjugate linoleic acid or CLA is a mixture of isomers that can be formed from 9 cis, 12 czs-octadecadienoic acid (linoleic acid) which can, theoretically, be autoxidized or alkali-isomerized into different conjugated geometric isomers including, but not limited to, 9 cis, 11 cis; 9 cis, 11 trans; 9 trans, 11 cis; 9 trans, 11 trans; 10 cis, 12 cis; 10 cis, 12 trans; 10 trans, 12 cis and 10 trans,
  • Dietary CLA particularly the 10 cis, 12 trans isomer, has direct effects on the proliferation and differentiation of cultured murine adipocytes Satroy, D. L. and Smith, S. B., J Nutr. (1999) 129:92-97; Park, Y., et al, (1999) Lipids 34:235-241.
  • Animals fed standard preparations of CLA consistently gain less weight than non- CLA fed controls. This can be a commercial disadvantage, in that it is often desirable to increase weight gain and rate of gain in animals raised to be food sources.
  • This effect of CLA can be seen in numerous papers including, for example, Wong, M. W., et al, Anticancer Research 17:987-994 (1997); Hayek, M.
  • AZOLE antifungal agent refers to agents having five-membered ring structures that contain two nitrogen atoms. More specifically the term encompasses imidazole and triazole agents such as: bifonazole, butoconazole, croconazole, clotrimazole, miconazole, fluconazole, econazole, fenticonazole, flutriazole, itraconazole, ketoconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, terconazole, and tioconazole.
  • imidazole and triazole agents such as: bifonazole, butoconazole, croconazole, clotrimazole, miconazole, fluconazole, econazole, fenticonazole, flutriazole, itraconazole, ketoconazole, omoconazole, oxiconazole, serta
  • AZOLE antifungal imidazole
  • P45014DM 14 ⁇ -demethylase
  • C-17 C-17
  • 20-lyase cholesterol side- chain cleavage enzyme
  • the 14- ⁇ -demethylase enzyme is involved in the sterol biosynthesis pathway and the synthesis of ergosterol from lanosterol.
  • Ergosterol is a necessary component of the fungal cytoplasmic membrane.
  • the antifungal activity (e.g., fungistatic or fungicidal) of this class of agents is therefore attributed to alterations of fungal cell membranes, resulting in increased membrane permeability, secondary metabolic effects, and growth inhibition.
  • some imidazole agents, such as miconazole seems to be attributed to direct physiochemical effects on the fungal cell membrane.
  • Ketoconazole which is the representative imidazole used in the examples presented herein, is structurally similar to miconazole and clotrimazole (Sonino N, (1987) New Eng. J. ofMed. 317: 812-817; Sohn C. A., (1982) Clin. Pharm., 1:211 -22A. KETO is an approved and marketed antifungal drug. The process of making ketoconazole is well known and described. This compound has good oral bioavailability. Ketoconazole is a white odorless, tasteless powder. It is stable at room temperature and requires acidity for dissolution and absorption. Ketoconazole is commercially available from Sigma Chemical Company (St Louis, MO).
  • Ketoconazole has been used as an oral and topical antimycotic agent (US Patent No. 6,080,744) with broad spectrum activity (against fungi, yeast and dermatophytes) and low toxicity.
  • ketoconazole is a cw-l-acetyl-4-[4- ⁇ [2- (2,4-dichlorophenyl)-2-(lH-imidazol-l-ylmethyl)-l,3-dioxolan-4-yl] methoxy ⁇ phenyl] piperazine.
  • the mechanism of action of ketoconazole is by inhibiting the synthesis of ergosterol in fungi and cholesterol in mammalian cells.
  • ergosterol is the main sterol in fungal cell membranes. It is to be understood that although the examples provided herein utilize KETO, the invention contemplates the use of any of the above-listed imidazole antifungal agents in combination with CLA, and the use of KETO for the experiments disclosed herein is not intended to limit the scope of the invention.
  • Ketoconazole is known to be a PPAR ⁇ activator which lowers blood glucose in diabetic animals and man.
  • PPAR ⁇ activators have shown clinical utility as anti- diabetic drugs. However, these drugs have a side effect of causing significant weight gain.
  • Differentiation of preadipocytes to adipocytes by ketoconazole has not been shown to be associated with the lipid accumulation normally seen with other PPAR ⁇ activators. Long term treatment with ketoconazole does not cause significant weight gain unlike other PPAR ⁇ activators.
  • Glucocorticoids like cortisol and cortisone are essential for abundant lipid accumulation during the differentiation of preadipocytes to adipocytes.
  • Ketoconazole may have properties similar to a cortisol anti-activator (Marin, US Patent # No. 5,849,740, 1998).
  • compositions of the invention comprise an effective amount of conjugated linoleic acid in combination with an imidazole antifungal agent.
  • the skin care compositions of the invention comprise from about 0.010% to about 40% of the active agents (i.e., conjugated linoleic acid and antifungal imidazole).
  • a skin care composition of the invention may comprise about 0.01%, 0.015%, 0.02%, 0.025%, 0.030%, 0.050%, 0.075%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.50%, 0.75%, 0.80%, 1%, 2.5%, 5%, 7.5%, 8.5%, 9.5%, 10%, 12%, 15%, 17.5%, 20%, 25%, 30%, 35%, 38% or 40% of each of the active ingredients (e.g. CLA and AZOLE) disclosed herein.
  • the invention provides a composition comprising from 0.025% - 10% conjugated linoleic acid (e.g.
  • the composition may comprise a single CLA isomer, such as a lOt, 12c-CLA or a 9, 11 isomer or mixture.
  • the composition can comprise a mixture of two or more CLA isomers.
  • a composition according to the invention could comprise a lOt, 12c-CLA isomer in combination with an 11, 13 isomer such as 11 -cis, 13-trans (1 lc, 13t).
  • both the CLA and AZOLE are each provided at a concentration of at least about 0.01%.
  • ketoconazole should be given at 100 mg - 1000 mg and the lOt, 12c-CLA isomer at 1000 mg- 5000 mg.
  • effective amount is an amount sufficient to provide cellulite reduction or prevention. It is accordingly an object of this invention to provide a composition which can reduce or eliminate cellulite or fat build-ups.
  • Cellulite as noted above, results from an accumulation of fatty materials and water imprisoned in a matrix made up of more or less water tight compartments. This matrix is comprised of elements of fundamental matter and more particularly of proteoglycons which are polymeric.
  • compositions of the invention can be formulated for oral or topical administration.
  • oral administration the composition is administered in a safe and effective dosage for cellulite prevention or reduction and for the treatment of obesity.
  • Oral administration of the composition results in decreased weight gain.
  • topical use the composition is presented in the form of a cream or oil for topical administration, usually in the form of a cream.
  • the methods of the invention encompass application of the composition used for local slimming and for fighting cellulite and for an entire body slimming effect if used orally.
  • composition according to the invention was conceived for fighting conditions of external appearance and figure, such as cellulite, general or local obesity, relaxing or ptosis of the skin and excessive secretion of fat (seborrhoea), which reveal profound bodily dysfunctions.
  • the compositions of the invention demonstrate slimming and "rejuvenating" effects on appearance.
  • positive results maybe obtained in terms of slimming and of reducing cellulite that has accumulated in a localized area of the body. That is, the composition is useful for preventing local fat and cellulite accumulation.
  • the skin becomes toned and fortified and the user feels no need, from an aesthetic point of view, to use another cream as a supplementing thereof.
  • compositions used in the present invention can comprise, consist of, or consist essentially of the essential elements and limitations of the invention described herein, as well any of the additional or optional ingredients, components, or limitations described herein.
  • references herein to a "patient” are intended to refer to human subjects with a desire to treat or prevent cellulite.
  • references herein to "animals” can be, but are not limited to, a rodent, a mammal (such as a bovine, an ovine, a caprine, a primate and a human), and an avian animal (such as a chicken, a duck, a turkey, and a quail).
  • compositions comprising CLA and an AZOLE are demonstrated by the synergistic effect that a combination comprising a lOt, 12c-CLA isomer and ketoconazole has on the stimulation of lipolysis (breakdown of triglycerides) by cultured human adipocytes.
  • a composition comprising a combination of CLA and Keto enhances the in vitro lipolysis relative to the amount of activity observed when either agent is used alone.
  • This new understanding permits one skilled in the art to produce compositions that comprise specific CLA isomers combined with an AZOLE that promote a desirable effect when administered while reducing or eliminating one or more undesirable effects.
  • the compositions of the invention may be administered orally or applied topically.
  • compositions of the present invention are exemplified herein by combinations comprising an indicated CLA isomer and ketoconazole, but it is understood that suitable compositions may also contain other CLA isomers, as well as other fatty acids, or alternative AZOLE agents.
  • the isomers can be extracted from natural sources or prepared using enzymatic or biological methods known to those skilled in the art. When making preparations of the invention, the source of the isomers is not critical, one should merely determine that the lOt, 12c isomers is provided in the composition at a percentage of at least about 0.01% to about 40% and that ketoconazole is present at a percentage of at least about 0.01% to about 40%.
  • the commercial CLA can be made from oils having at least 50% linoleic acid and which can contain 95%. linoleic acid or more.
  • the cost of CLA isomers increases with increasing purity.
  • Bulk conjugated linoleic acid isomers in a significantly purified form (98%+pure) are commercially available from Matreya, Inc. (Pleasant Gap, Pa.).
  • Matreya, Inc. Pleasant Gap, Pa.
  • the compositions can comprise a (lOt, 12c) isomer or a (9c, 1 It) isomer of
  • CLA isomer mixed with other CLA isomers preferably the composition comprises only a single CLA isomer.
  • the isomers are heat stable and can be used as is, or dried and powdered. Purified isomers of CLA are also commercially available from Matreya.
  • an effective amount of a CLA/AZOLE composition prepared as a topical skin care composition is administered to the patient.
  • CLA is a natural food ingredient and antifungal imidazole agents are commonly used ingredients in anti-fungal topical applications both of the active agents of the disclosed compositions are relatively non-toxic and the amount of CLA/AZOLE that can be administered is not critical as long as it is enough to be effective to achieve the desired outcome noted herein.
  • the methods of the present invention may take several embodiments.
  • the CLA/AZOLE mixture is administered in a pharmaceutical or cosmetic composition containing a safe and effective dose of the CLA and AZOLE.
  • a pharmaceutically or cosmetically acceptable carrier may additionally be provided.
  • the formulations of the invention comprise a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier is intended a carrier that is conventionally used in the art to facilitate the storage, administration, and/or the healing effect of the therapeutic ingredients.
  • a carrier may also reduce any undesirable side effects of the CLA and/or the AZOLE agent present in the composition.
  • a suitable carrier should be stable, i.e., incapable of reacting with other ingredients in the formulation. It should not produce significant local or systemic adverse effects in recipients at the dosages and concentrations employed for treatment. Such carriers are generally known in the art.
  • Suitable carriers for this invention are those conventionally used large stable macromolecules such as albumin, for example, human serum albumin, gelatin, collagen, polysacchari.de, monosaccharides, polyvinyl-pyrrolidone, polylactic acid, polyglycolic acid, polymeric amino acids, fixed oils, ethyl oleate, liposomes, glucose, sucrose, lactose, mannose, dextrose, dextran, cellulose, sorbitol, polyethylene glycol (PEG), and the like.
  • Slow- release carriers such as hyaluronic acid, may also be suitable. See particularly Prisell et al (1992) Int. J. Pharmaceu. 85:51-56, and U.S. Patent No.
  • compositions include, but are not limited to, pharmaceutically acceptable agents that modify isotonicity including water, salts, sugars, polyols, amino acids, and buffers.
  • suitable buffers include phosphate, citrate, succinate, acetate, and other organic acids or their salts and salts that modify the tonicity such as sodium chloride, sodium phosphate, sodium sulfate, potassium chloride, and can also include the buffers listed above.
  • a cosmetically acceptable vehicle is comprised either of water or of a water/solvent blend.
  • the solvent is optimally chosen from propylene glycol, ethanol, butylene glycol, and polyethylene glycols of various molecular weights.
  • Vehicles other than 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.
  • 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.
  • the cosmetically acceptable vehicle will usually form from 5% to 99.9%, preferably from 25% to 80% by weight of the emulsion, and can, in the absence of other cosmetic adjuncts, form the balance of the composition.
  • the compositions used in the present invention also contain a dermatologically acceptable carrier.
  • dermatologically-acceptable carrier means that the carrier is suitable for topical application to the skin, has good aesthetic properties, is compatible with the active ingredients of the present invention and any other components, and will not cause any untoward safety or toxicity concerns.
  • a safe and effective amount of carrier is from about 50% to about 99.99%, preferably from about 99.9% to about 80%, more preferably from about 98% to about 90%, most preferably from about 95% to 90% of the composition.
  • the carrier can be in a wide variety of forms.
  • emulsion carriers including, but not limited to, oil-in-water, water-in-oil, water-in-oil-in-water, and oil- in-water-in-silicone emulsions, are useful herein. These emulsions can cover a broad range of viscosities, e.g., from about 100 cps to about 200,000 cps. These emulsions can also be delivered in the form of sprays using either mechanical pump containers or pressurized aerosol containers using conventional propellants. These carriers can also be delivered in the form of a mousse.
  • suitable topical carriers include anhydrous liquid solvents such as oils, alcohols, and silicones (e.g., mineral oil, ethanol, isopropanol, dimethicone, cyclomethicone, and the like); aqueous-based single phase liquid solvents (e.g., hydro-alcoholic solvent systems); and thickened versions of these anhydrous and aqueous-based single phase solvents (e.g., where the viscosity of the solvent has been increased to form a solid or semi-solid by the addition of appropriate gums, resins, waxes, polymers, salts, and the like).
  • anhydrous liquid solvents such as oils, alcohols, and silicones (e.g., mineral oil, ethanol, isopropanol, dimethicone, cyclomethicone, and the like)
  • aqueous-based single phase liquid solvents e.g., hydro-alcoholic solvent systems
  • thickened versions of these anhydrous and aqueous-based single phase solvents e.
  • topical carrier systems useful in the present invention are described in the following four references all of which are incorporated herein by reference in their entirety: "Sun Products Formulary” Cosmetics & Toiletries, Vol. 105, pp. 122-139 (December 1990); “Sun Products Formulary", Cosmetics & Toiletries, Vol. 102, pp. 117-136 (March 1987); U.S. Pat. No. 4,960,764 to Figueroa et al, issued Oct. 2, 1990; and U.S. Pat. No. 4,254,105 to Fukuda et al, issued Mar. 3, 1981.
  • the carriers of the skin care compositions can comprise from about 50% to about 99% by weight of the compositions used in the present invention, preferably from about 75% to about 99%, and most preferably from about 85% to about 95%.
  • Preferred cosmetically and/or pharmaceutically acceptable topical carriers include hydroalcoholic systems and oil-in-water emulsions.
  • the carrier is a hydro- alcoholic system
  • the carrier can comprise from about 0% to about 99% of ethanol, isopropanol, or mixtures thereof, and from about 1% to about 99% of water. More preferred is a carrier comprising from about 5% to about 60% of ethanol, isopropanol, or mixtures thereof, and from about 40% to about 95% of water.
  • a carrier comprising from about 20% to about 50% of ethanol, isopropanol, or mixtures thereof, and from about 50% to about 80% of water.
  • the carrier is an oil-in-water emulsion
  • the carrier can include any of the common excipient ingredients for preparing these emulsions.
  • compositions used in the present invention may optionally comprise additional materials including slimming agents as well as additional actives useful in providing cellulite control.
  • a second anti-cellulite agent can be included in the composition.
  • these agents are phosphodiesterase inhibitors (e.g., xanthine derivatives such as theophyUine, caffeine, theobromine or salts thereof such as aminophylline) and certain oleosoluble vegetable extracts, including, principally, those of climbing ivy (Hedera helix), arnica (Arnica montana), rosemary (Rosmarinus officinalis N), marigold (Calendula officinalis), sage (Salvia officinalis N), ginseng (Panax ginseng), St.
  • phosphodiesterase inhibitors e.g., xanthine derivatives such as theophyUine, caffeine, theobromine or salts thereof such as aminophylline
  • certain oleosoluble vegetable extracts including, principally
  • a Xanthines can be employed at a concentration such that it is present in a proportion from about 0.05% to about 20%> by weight of the composition.
  • compositions used in the present invention may optionally comprise additional skin actives.
  • Non-limiting examples of such skin actives include hydroxy acids such as salicylic acid; desquamatory agents such as zwitterionic surfactants; sunscreens such as 2-ethylhexyl-p-methoxycinnamate, 4,4'-t-butyl methoxydibenzoyl-methane, octocrylene, phenyl benzimidazole sulfonic acid; sun-blocks such as zinc oxide and titanium dioxide; anti-inflammatory agents; corticosteroids such as hydrocortisone, methylprednisolone, dexamethasone, triamcinolone acetconide, and desoxametasone; anesthetics such as benzocaine, dyclonine, lidocaine and tetracaine; antipruitics such as camphor, menthol, oatmeal (colloidal), pramoxine, benzyl alcohol, phenol and resorcinol; anti-oxidants/radical
  • Preferred skin actives include hydroxy acids such as salicylic acid, sunscreen, antioxidants and mixtures thereof.
  • Other conventional skin care product additives may also be included in the compositions used in the present invention.
  • urea urea, guanidine, glycerol, petrolatum, mineral oil, sugar esters and polyesters, polyolefins, methyl isostearate, ethyl isostearate, cetyl ricinoleate, isononyl isononanoate, isohexadecane, lanolin, lanolin esters, cholesterol, pyrrolidone carboxylic acid/salt (PCA), trimethyl glycine (betaine), tranexamic acid, amino acids (e.g., serine, alanine), panthenol and its derivatives, collagen, hyaluronic acid, elastin, hydrolysates, primrose oil, jojoba oil, epidermal growth factor, soybean saponins, mucopolysaccharides, and mixtures thereof may be used.
  • PCA pyrrolidone carboxylic acid/salt
  • betaine trimethyl glycine
  • tranexamic acid amino
  • compositions used in the present invention are generally prepared by conventional methods such as are known in the art of making topical compositions. Such methods typically involve mixing of the ingredients in one or more steps to a relatively uniform state, with or without heating, cooling, application of vacuum, and the like.
  • Non-limiting examples of the product form can be a gel, emulsion, lotion, cream, ointment, solution, liquid, etc.
  • the methods of the present invention are useful for preventing cellulite, especially in the subcutaneous, dermis and epidermis tissues of mammalian skin.
  • the methods of the present invention involve topically applying to the skin an effective amount of the skin care composition of the present invention.
  • the amount of the composition which is applied, the frequency of application and the period of use will vary widely depending upon the concentration of CLA (e.g. 10-trans, ⁇ 2-cis conjugated linoleic acid), AZOLE (e.g., ketoconazole) and/or other component of a given composition and the degree of cellulite fading desired.
  • CLA e.g. 10-trans, ⁇ 2-cis conjugated linoleic acid
  • AZOLE e.g., ketoconazole
  • the skin care compositions used in the present invention can be administered by chronic topical application.
  • chronic topical application is meant continued topical application of the composition over an extended period during the subject's lifetime, preferably for a period of at least about one week, more preferably for a period of at least about two weeks, even more preferably for a period of at least one month, even more preferably for at least about three months, even more preferably for at least about six months, and more preferably still for at least about one year. While benefits are obtainable after various maximum periods of use (e.g., five, ten or twenty years), it is preferred that chronic application continue throughout the subject's lifetime to maintain and/or increase the benefits achieved.
  • compositions used in the present invention can be employed to provide a skin appearance and/or feel benefit.
  • Quantities of the present compositions which are typically applied per application are, in mg composition/cm.sup.2 skin, from about 0.1 mg/cm.sup.2 to about 10 mg/cm.sup.2.
  • the method of treating cellulite is preferably practiced by applying a composition in the form of a skin lotion, cream, gel, cosmetic, or the like which is intended to be left on the skin for some aesthetic, prophylactic, therapeutic or other benefit (i.e., a "leave-on" composition).
  • a composition in the form of a skin lotion, cream, gel, cosmetic, or the like which is intended to be left on the skin for some aesthetic, prophylactic, therapeutic or other benefit (i.e., a "leave-on" composition).
  • a composition in the form of a skin lotion, cream, gel, cosmetic, or the like which is intended to be left on the skin for some aesthetic, prophylactic, therapeutic or other benefit
  • a “leave-on" composition After applying the composition to the skin, it is preferably left on the skin for a period of at least about 15 minutes, more preferably at least about 30 minutes, even more preferably at least about 1 hour, most preferably for at least several hours, e.g., up to about
  • the patch can be occlusive, semi-occlusive or non-occlusive.
  • a composition comprising lb-trans, 12-cis conjugated linoleic acid and ketoconazole could be contained within a patch that is contacted with the skin; or alternatively, a composition comprising CLA and AZOLE could be applied to the skin prior to the application of a patch.
  • the patch can also include additional actives such as chemical initiators for exothermic reactions such as those described in PCT application WO 9701313 to Burkett et al.
  • the patch is applied at night as a form of night therapy.
  • the preferred xanthine employed in the invention is caffeine and/or theophyUine due to their availability and optimum efficacy.
  • Caffeine and theophyUine can be, and preferably are naturally-derived, in order to keep with a "natural" character of the inventive compositions.
  • the xanthine is employed in the inventive method preferably in an amount of at least 0.05%, generally in the amount of from 0.05% to 20%, preferably in the amount of from 0.10% to 10%, optimally in the amount of from 0.5% to 3.0% by weight of the composition in order to maximize efficacy at optimum cost.
  • inventive compositions contain the L- form of an hydroxy acid.
  • the amount of the hydroxy acid component present in the composition according to the invention is from 1.5% to 20%, more preferably from 1.5% to 15%, and most preferably from 3.0% to 12.0% 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 benefit agents other than emollients and other than ingredients that merely improve the physical characteristics of the composition.
  • general examples include sunscreens, tanning agents, skin anti-wrinkling agents, anti-inflammatory agents, skin lighteners and moisturizers.
  • Sunscreens include those materials commonly employed to block ultraviolet light.
  • Illustrative compounds are the derivatives of PABA, and cinnamate.
  • octyl methoxycinnamate and 2-hydroxy-4-methoxybenzophenone also known as oxybenzone
  • Octyl methoxy-cinnamate and 2-hydroxy-4- methoxy benzophenone are commercially available under the trademarks, Parsol MCX and Benzophenone-3, respectively.
  • the exact amount of sunscreen employed in the emulsions can vary depending upon the degree of protection desired from the sun's UV radiation.
  • Suitable anti-inflammatory compounds include but are not limited to rosmarinic acid, glycyrrizinate derivatives, ⁇ bisabolol, azulene and derivatives thereof, asiaticoside, sericoside, ruscogenin, escin, esculin, quercetin, rutin, betulinic acid and derivatives thereof, catechin and derivatives thereof.
  • Suitable vasoactive compounds include but are not limited to papaverine, yohimbine, visnadin, khellin, bebellin, nicotinate derivatives.
  • Surfactants which are also sometimes designated as emulsifiers, may be incorporated into the cosmetic compositions of the present invention.
  • Surfactants can comprise anywhere from about 0.5% to about 30%, preferably from about 1% to about 15%) by weight of the total composition.
  • Surfactants may be cationic, nonionic, anionic, or amphoteric in nature and combinations thereof may be employed.
  • nonionic surfactants are alkoxylated compounds based upon fatty alcohols, taffy acids and sorbitan. These materials are available, for instance, from the Shell Chemical Company under the "Neodol” designation. Copolymers of polyoxypropylene-polyoxyethylene, available under the Pluronic trademark sold by the BASF Corporation, are sometimes also useful. Alkyl polyglycosides available from the Henkel Corporation similarly can be utilized for the purposes of this invention.
  • Anionic-type surfactants may include fatty acid soaps, sodium lauryl sulphate, sodium lauryl ether sulphate, alkyl benzene sulphonate, mono and/or dialkyl phosphates and sodium fatty acyl isethionate.
  • Amphoteric surfactants include such materials as dialkylamine oxide and various types of betaines (such as cocoamido propyl betaine).
  • 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 from about 0.5% to 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 maybe employed such as xanthan, carrageenan, gelatin, karaya, pectin and locust bean 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.
  • Cellulosic derivatives may also be employed, e.g., hydroxypropyl cellulose (Klucel HI.RTM.).
  • Suitable preservatives include alkyl esters of p- hydroxybenzoic acid, hydantoin derivatives, propionate salts, and a variety of quaternary ammonium compounds.
  • Particularly preferred preservatives of this invention are methyl paraben, propyl paraben, imidazolidinyl urea, sodium dehydroxyacetate and benzyl alcohol.
  • Preservatives will usually be employed in amounts ranging from about 0.5%> to 2%> by weight of the composition.
  • Powders may be incorporated into the cosmetic composition employed in the 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.
  • adjunct minor components may also be incorporated into the cosmetic compositions.
  • 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.
  • the method of the present invention is useful for reducing or preventing the appearance of cellulite, for improving the firmness and elasticity of skin and generally to enhance the quality and flexibility of skin.
  • Type I collagenase was obtained from Worthington Biochemical (Lakewood, NJ).
  • Bovine serum albumin (BSA), dexamethasone, biotin, pantothenate, Krebs-Ringer buffer, isoproterenol, and isobutyl methylxanthine were purchased from Sigma (St. Louis, MO).
  • Human recombinant insulin was obtained from Boehringer Mannhaim (Indianapolis, IN).
  • the fetal bovine serum was from HyClone (Logan, Utah).
  • DMEM Dublecco Modified Eagles Medium
  • PBS phosphate buffered saline
  • adipocyte precursor cells preadipocytes
  • the adipocyte precursor cells are isolated from subcutaneous adipose tissue as described. The plates are kept at 37°C with 5% CO 2 until ready for use. Differentiation into adipocytes should be initiated immediately. If cells are to be maintained as preadipocytes, they should be fed with preadipocyte medium every other day. Preadipocytes are flat, phase-dark spindle-shaped cells. The cells have a similar appearance in culture to fibroblasts or smooth muscle cells. Greater than 80% of the preadipocytes will differentiate to adipocytes using differentiation medium (DM-2/10). The differentiation efficiency varies depending on the donor.
  • DM-2/10 differentiation medium
  • Tissue preparation The subcutaneous adipose was acquired from elective surgeries (liposuction) with the patient's consent as approved by the hospital histitution Review Boards. The tissues used were from female patients. Other than obesity in some instances, the patients were in good health; no diabetes or other complications were reported. Tissues isolated from either the abdomen or the thigh/hip regions were transported to the laboratory in saline solution and in separate containers within two hours post-surgery. The tissue was washed several times with 2 volumes of Krebs-
  • Ringer-Bicarbonate (KRB; Sigma Chemical Co., St. Louis, MO) to remove blood.
  • the tissue was then digested with one volume of collagenase type I (1 g/liter of KRB with 1% BSA) for 60 minutes at 37° C with intermittent shaking.
  • the floating adipocytes were separated from the stromal- vascular fraction by centrifugal force (300 g) for 5 minutes.
  • Preadipocytes are differentiated into adipocytes. The plates are kept at 37°C with 5% CO 2 until ready for use. The adipocytes should be fed with adipocyte medium (AM-1) every 3 days. The adipocytes should remain healthy and responsive for at least three weeks. Adipocytes are rounded, lipid-filled cells. Cultured adipocytes contain multiple vesicles termed "locules". These locules are the site of lipid storage and can be visualized by counterstaining with Oil red O.
  • adipocytes Differentiation of adipocytes: The stromal cells were trypsinized and plated in multiple- well plates at 40,000 cells/cm for 16 hours to allow attachment. The medium was then changed to Dulbecco's modified Eagle's -Ham's F-10 medium (vol/vol, 1:1) supplemented with 3% fetal bovine serum, 15mM HEPES (pH 7.4), biotin (33 ⁇ M), pantothenate (17 ⁇ M, Sigma, MO), human recombinant insulin (100 nM, Boerhinger Mannheim, IN), dexamethasone (1 ⁇ M), l-methyl-3-isobutylxanthine (0.2 mM), and a PPAR ⁇ agonist (1 ⁇ M) for 3 days. During the remaining 9-10 day adipocyte differentiation period, the cells were fed every 3 days with the same medium without l-methyl-3-isobutylxanthine and PPAR ⁇ agonist supplementation.
  • Lipolysis Assay Day 21 differentiated human adipocytes plated in 96 well plates (Falcon, etc.) generated as described (Halvorsen et al, Metabolism. 50(4):407-13, 2001), were used. The medium was removed completely and 150 ⁇ l of the test compound resuspended in KRB added to each well. The plates were incubated at
  • SV Stromal Vascular cells were seeded at a density of 3 x 10 4 /cm 2 and continuously treated with increasing concentrations (3, 10, or 30 uM) of either linoleic acid, cis-9, tran-11 CLA, or trans-10, cis-12 CLA.
  • a set of control cultures received vehicle (BSA) plus TZD.
  • AU cultures received differentiation media (days 1-3), adipocyte media (days 4-9), and low-glucose ( ⁇ 5mM) adipocyte media (days 10-12) prior to measuring de novo lipogenesis.
  • C-labeled glucose incorporation into the lipid fraction of the cultures was measured for 2 h and, following lipid extraction, the radioactivity in the lipid fraction was determined by scintillation counting.
  • the lipolytic activities of 10-trans, 12-cis conjugated linoleic acid, ketoconazole, Isoproterenol and Isobutylmethylxanthine were evaluated in the above-described assay. The results are summarized in Table 1. The data summarize the activity of the various compounds at different concentrations relative to the level of lipolysis observed in the control sample. The data indicate that CLA stimulates lipolysis (e.g. the breakdown of stored triglycerides into fatty acids and glycerol) in human adipocytes.
  • Table 2 summarizes the relative activity level (e.g., stimulation of glycerol release expressed as fold increase over an appropriate control) of combinations comprising CLA and KETO compared to the activity of other know stimulators of lipolysis in cultured human adipocytes.
  • Lipolysis experiments were performed on adipocytes isolated from 2 different patients (LOl 1701 and LOl 1901H). The effect of 10-trans, 12-cis conjugated linoleic acid (CLA) and ketoconazole (KETO) either alone or in the combinations defined in Table 3 were evaluated. All of the values used to define the compositions listed in table 3 refer to ⁇ M concentrations.
  • the lipolysis assay was done according to the above-described method and the results are graphically displayed in Figures 1 A and IB.
  • KETO consistently release higher levels of glycerol (e.g., lipolysis). More specifically, stimulation with combinations comprising combination #8 (100 ⁇ M CLA and 10 ⁇ M KETO), combination #9 (10 ⁇ M CLA and 100 ⁇ M KETO) and combination #13 (100 ⁇ M CLA and 1 ⁇ M KETO) resulted in the highest levels of glycerol release.
  • combination #8 100 ⁇ M CLA and 10 ⁇ M KETO
  • combination #9 10 ⁇ M CLA and 100 ⁇ M KETO
  • combination #13 100 ⁇ M CLA and 1 ⁇ M KETO
  • Mature adipocytes plated in 96-well plates were washed once with Krebs- Ringer buffer supplemented with lg / liter glucose and were incubated with or without lipolytic reagent (120 ⁇ l per well) for 3 hours at 37°C.
  • the conditioned media 100 ⁇ l/ well) were removed and the amount of released glycerol determined using the triglyceride reagent A (GPO-Trinder, Sigma, St. Louis, MO) following manufacturer's instruction.
  • Lipolysis was assayed by measuring the glycerol released from the mature adipocytes obtained from two different sources (e.g., LOl 1901 H and LOl 1701) in response to the control agent Isoproterenol (Iso), conjugated linoleic acid (CLA) alone, ketoconazole (KETO) alone, and compositions comprising CLA and KETO in combination.
  • Isoproterenol Isoproterenol
  • CLA conjugated linoleic acid
  • KETO ketoconazole
  • Example 4 Effect of CLA and Ketoconazole on Lipolysis in Cultured Human Adipocytes
  • compositions comprising CLA and ketoconazole (KETO) on lipolysis were also evaluated in the above-described assay, using human adipocytes obtained from subcutaneous adipose tissue removed during liposuction surgery of either the abdomen (light bars) or the thigh hip region (dark bars) of female patients.
  • the results presented in figure 4 indicate that a combination of CLA and KETO is more effective in stimulating lipolysis than either compound alone.
  • the effect of linoleic acid, cis-9, trans-11 CLA, and trans-10, cis-12 CLA on lipogenesis in cultured human adipocytes was evaluated using the above-described lipogenesis assay. More specifically, lipogenesis was assayed by determining the level of C-glucose incorporation into total cellular lipid [(pmol/(L 10 )] during de novo lipogenesis of cultured human adipocytes. The results are presented in Figure 3. Data are expressed as percentage of vehicle (BSA) controls. Means (+_ SEM, n+6) not sharing a letter differ, p ⁇ 0.05. The data indicate that CLA blocks the uptake of fat in the form of triglycerides by cultured human adipocytes.
  • compositions of the present invention describe two alternative examples of a skin cream incorporating the compositions of the present invention.
  • the compositions are formed by combining and mixing the ingredients of each column using conventional technology and then applying about 0.5g to about 50 g of the mixture to the skin.

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Abstract

La présente invention concerne des compositions et des méthodes permettant de prévenir et/ou de traiter chez les mammifères la cellulite qui se caractérise par une accumulation de tissus adipeux. La composition de soin pour la peau comprend de l'acide linoléique conjugué (CLA), un agent antifongique AZOLE et un excipient pharmaceutiquement acceptable. Dans une forme de réalisation particulière, la composition comprend des quantités efficaces d'acide linoléique conjugué 10-trans, 12-cis, de kétoconazole et d'un excipient dermatologiquement acceptable. Les méthodes de prévention et/ou de traitement consistent à appliquer de manière topique une composition comprenant des quantités non dangereuses et efficaces d'acide linoléique conjugué en combinaison avec un agent antifongique imidazole. Les compositions selon l'invention améliorent l'aspect de la peau du fait qu'elles favorisent la lipolyse et/ou qu'elles inhibent la lipogénèse de novo par les adipocytes localisés sur des régions ciblées de la peau du mammifère.
PCT/US2002/011079 2001-07-11 2002-04-08 Compositions permettant de reduire la cellulite ou de prevenir son apparition chez les mammiferes WO2003006009A1 (fr)

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US20110268688A1 (en) * 2008-06-25 2011-11-03 Mccarthy James Timothy Method and compositions for improving skin and body appearance
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US11135139B2 (en) 2013-03-15 2021-10-05 Belaj Innovations Llc Cosmetic compositions and uses thereof
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US10299994B2 (en) 2013-03-15 2019-05-28 Mary Kay Inc. Cosmetic compositions and uses thereof
EP2968107A4 (fr) * 2013-03-15 2016-08-17 Mary Kay Inc Compositions cosmétiques et leurs utilisations
US11723845B2 (en) 2013-03-15 2023-08-15 Mary Kay Inc. Cosmetic compositions and uses thereof
WO2014195872A1 (fr) 2013-06-04 2014-12-11 Vyome Biosciences Pvt. Ltd. Particules enrobées et compositions les comprenant
US12286413B2 (en) 2014-08-07 2025-04-29 Mayo Foundation For Medical Education And Research Compounds and methods for treating cancer
US11325892B2 (en) 2014-08-07 2022-05-10 Mayo Foundation For Medical Education And Research Compounds and methods for treating cancer
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US11039990B2 (en) * 2018-05-04 2021-06-22 Johnson & Johnson Consumer Inc. Cleansing compositions
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US11202737B2 (en) 2018-05-04 2021-12-21 Johnson & Johnson Consumer Inc. Cleansing compositions
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