VEGFC PRODUCTION PROMOTER

Abstract

The purpose of this invention is to find a novel fragrance component effective for prevention or control of swelling, lymphedema, skin aging or obesity from the viewpoint of activation of lymph vessel function by promotion of the expression of VEGFC. This invention provides a swelling improving agent, lymph vessel activator and VEGFC production promoter characterized by containing one or a plurality of components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

Claims

1.-6. (canceled)

7. A method of promoting VEGFC production comprising administering a formulation comprising one or more components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

8. A method of promoting lymphangiogenesis and/or lymphatic function comprising administering a formulation comprising one or more components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

9. A method of ameliorating swelling, comprising a formulation comprising one or more components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

10. A method of ameliorating lymphedema comprising a formulation comprising one or more components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

11. A method of ameliorating wrinkles comprising administering a formulation comprising one or more components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

12. A method of ameliorating obesity comprising administering a formulation comprising one or more components selected from the group consisting of rose oxide, citronellyl acetate and niaouli oil.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0040] FIG. 1 is a graph showing the VEGFC-inducing activity of rose oxide, citronellyl acetate and niaouli oil.

MODE FOR CARRYING OUT THE INVENTION

[0041] Definitions:

[0042] The term “swelling” in the present description is used in the same context as “edema”, and refers to a state observed in subcutaneous tissue in which tissue fluid or lymphatic fluid accumulates in cells, intercellular space or body cavities due to some cause. Moreover, the term “lymphedema” in the present description refers to a state in which tissue fluid accumulates in distal regions due to occlusion of lymph vessels.

[0043] As is described in detail in the subsequent examples, screening for agents that induce a lymph vessel activating factor in the form of VEGFC was carried out by allowing various types of candidate aromatics to act on human epidermis-derived cultured cells in the form of HaCaT cells, quantifying VEGFC, and selecting aromatics having VEGFC-inducing activity as active ingredients by using the amount of VEGFC as an indicator of VEGFC-inducing activity.

[0044] As a result, rose oxide, citronellyl acetate and niaouli oil were found to induce VEGFC.

[0045] * Rose Oxide

[0046] Rose oxide is an organic compound that has the chemical structure indicated below and has two asymmetric carbons within a molecule thereof.

##STR00001##

Rose oxide has a sweet fragrance like that of roses, and is used as a pyran-based, monoterpene-based aromatic in cosmetics and soaps. It can be obtained from roses, rose oil, apricots and peppermint in nature, and has a plurality of geometrical and optical isomers. Industrially, it is obtained in equal amounts of the cis-form and trans-form by reducing allyl hydroperoxides, which are obtained by photooxidation of citronellol, to obtain a diol followed by closing the ring with sulfuric acid. Commercial products available as aromatics consist primarily of the cis racemic form.

[0047] * Citronellyl Acetate

[0048] Citronellyl acetate is an organic compound represented by the following chemical formula that is in the form of a colorless, clear liquid at normal temperature.

##STR00002##

It is used as an aromatic of foods and cosmetics since it has a unique fragrance that resembles that of roses or lavender. In nature, it is contained in essential oils such as citronella oil or geranium oil, and is synthesized industrially from citronellol.

[0049] * Niaouli Oil

[0050] Niaouli oil is an essential oil collected from the leaves or branches of Melaleuca virdiflora, a plant of the Myrtaceae family found in the Pacific region such as in Indonesia, Australia and New Zealand, and on Madagascar Island in the Indian Ocean. It has bactericidal action and is used in cosmetics, soaps or bath additives and the like.

[0051] The aforementioned aromatic components according to the present invention are extremely useful for promoting the formation and function of lymph vessels. Symptoms accompanying lymph vessel dysfunction include not only swelling and lymphedema, but also photoaging (such as wrinkle formation) of skin accompanying exposure to ultraviolet rays and obesity. The agent according to the present invention is effective for preventing or inhibiting swelling and lymphedema along with photoaging of the skin accompanying exposure to ultraviolet rays and obesity. Moreover, the aforementioned aromatic components are also thought to be useful in the treatment of congenital lymphedema.

[0052] Photoaging of the skin refers to a change in the appearance and function of skin typically observed as a result of repeated exposure to sunlight. Constituent elements of sunlight in the form of ultraviolet (UV) rays, and particularly ultraviolet B rays (referred to as UVB rays having a wavelength of 290 nm to 320 nm), are the primary cause of photoaging. The exposure level of UVB required to cause photoaging is currently not known. However, repeated exposure to UVB at a level that causes erythema or sunburn normally leads to photoaging. In clinical terms, photoaging can be identified by, for example, rough skin, wrinkle formation, colored spots, pale color, sagging skin, onset of telangiectasia, mole formation, onset of purpura, increased susceptibility to wounds, atrophy, appearance of regions of fibrotic pigment removal and the onset of premalignant and malignant tumors. Photoaging usually occurs in skin that has been routinely exposed to sunlight, such as that of the face, ears, head, neck and hands.

[0053] The dosage, administration form and dosage form of the VEGFC production promoter, lymphangiogenesis and/or lymphatic function promoter, swelling ameliorant, lymphedema ameliorant, wrinkle ameliorant or obesity ameliorant of the present invention (to also be referred to as the “present agent”) can be suitably determined corresponding to the purpose of use thereof. For example, the aromatic component in the present agent is typically incorporated at 0.0001% by weight to 0.1% by weight, preferably at 0.001% by weight to 0.01% by weight, and optimally at 0.005% by weight to 0.01% by weight based on the total weight of the drug. Although there are no particular limitations on the administration form of the present agent, and may be administered orally, parenterally, externally or by inhaling and the like, it is preferably a skin external agent from the viewpoint of being able to act both by inhaling and transcutaneously. Examples of dosage forms include cosmetics such as perfumes, colognes, shampoos, rinses, skin care products, body shampoos, body rinses, body powders, air fresheners, deodorants, bath additives, lotions, creams, soaps, toothpastes or aerosol products, and other forms commonly used in aromatics. Moreover, the present agent can also be used in pharmaceuticals such as inhalants.

[0054] In addition, the present agent can also suitably incorporate as necessary other components in addition to the aforementioned essential components such as components normally used in foods or pharmaceuticals such as external skin agents in the manner of skin whitening agents, moisturizers, antioxidants, oily components, ultraviolet absorbers, surfactants, thickeners, alcohols, powdered components, coloring agents, aqueous components, water or various types of skin nutrients.

[0055] In addition, other components can also be suitably incorporated, examples of which include sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, hot water extracts such as caffeine, tannin, verapamil, tranexamic acid and derivatives thereof, licorice extract, glabridin or quince fruit extract, various types of natural herbs, drugs such as tocopherol acetate, glycyrrhizic acid and derivatives thereof, and salts thereof, skin whitening agents such as vitamin C, magnesium ascorbyl phosphate, ascorbic acid glucoside, albumin or kojic acid, sugars such as glucose, fructose, mannose, sucrose or trehalose, and vitamin A such as retinoic acid, retinol, retinol acetate or retinol palmitate.

EXAMPLES

[0056] Human-derived cultured cells in the form of HaCaT cells (German Cancer Research Center, Heidelberg, Germany) were cultured in DMEM medium (Gibco Low Glucose DMEM™, Life Technologies Inc.) containing 10% FBS (Standard Fetal Bovine Serum, Hyclone Laboratories Inc.) and disseminated in a 24-well plate to 15×10.sup.4 cells/well. The medium was replaced with serum-free DMEM 24 hours after dissemination followed by culturing for 24 hours. Subsequently, each aromatic dissolved in ethanol was added (aromatic final concentration: 0.001% to 0.01%). Ethanol was used as a control. The culture supernatant was recovered after culturing for 48 hours and the amount of VEGFC in the culture supernatant was quantified in accordance with the protocol of an ELISA kit (Quanikine® Human VEGF-C Immunoassay (R&D Systems Inc., Minneapolis, Minn.)). In addition, alamar blue (TREK Diagnostic Systems, Inc., Cleveland, US) was added to the cells after recovering the culture supernatant followed by measuring the number of cells based on their fluorescence intensity (Ex: 544 nm, Em: 590 nm). The amount of VEGFC in the supernatant was divided by fluorescence intensity, and the relative VEGFC-inducing activity per cell based on a value of 100 for the control (ethanol) is shown in FIG. 1. The synthetic aromatics of rose oxide and citronellyl acetate along with the natural aromatic, niaouli oil (supplier: Biolandes Inc.), demonstrated VEGFC-inducing activity as shown in FIG. 1 without demonstrating cytotoxicity at each concentration.

[0057] Formulation Examples

[0058] Although the following indicates formulation examples of the VEGFC production promoter of the present invention, carrying out the present invention is not limited to the following examples.

[0059] Beauty Wash

TABLE-US-00001 Formula (wt %) (1) Glycerin 2.0 (2) Dipropylene glycol 2.0 (3) PEG-60 hydrogenated castor oil 0.3 (4) Trimethylglycine 0.1 (5) Preservative As suitable (6) Chelating agent As suitable (7) Dye As suitable (8) VEGFC production promoter of present 0.05 invention: Rose oxide (9) Purified water Balance

[0060] Beauty Wash

TABLE-US-00002 Formula (wt %) (1) Alcohol 30.0 (2) Butylene glycol 4.0 (3) Glycerin 2.0 (4) PPG-13-decyltetradeceth-24 0.3 (5) Octyl methoxycinnamate 0.1 (6) Menthol 0.2 (7) Tocopherol acetate 0.1 (8) Chelating agent As suitable (9) Dye As suitable (10) VEGFC production promoter of present 0.1 invention: Niaouli oil (11) Purified water Balance

[0061] Milky Lotion

TABLE-US-00003 Formula (wt %) (1) Stearic acid 2.0 (2) Cetyl alcohol 1.5 (3) Vaseline 4.0 (4) Squalane 5.0 (5) Glycerol tri-2-ethylhexanoate 2.0 (6) Sorbitan monooleate 2.0 (7) Dipropylene glycol 5.0 (8) PEG1500 0.3 (9) Triethanolamine 0.1 (10) Preservative As suitable (11) VEGFC production promoter of present 0.01 invention: Citronellyl acetate (12) Purified water Balance

[0062] Milky Lotion

TABLE-US-00004 Formula (wt %) (1) Ethyl alcohol 10.0 (2) Cyclomethicone 0.1 (3) Butylene glycol 5.0 (4) Dimethicone 3.0 (5) Glycerin 0.1 (6) Menthol 1.0 (7) Trimethylsiloxycinnamic acid 0.1 (8) Caffeine 1.0 (9) Trimethylglycine 1.0 (10) Xanthan gum 0.001 (11) Hydroxyethyl cellulose 0.1 (12) Fermented soybean extract 1.0 (13) Lauryl betaine 0.5 (14) Carbomer 0.2 (15) Chelating agent As suitable (16) Para-hydroxybenzoate As suitable (17) Benzoic acid As suitable (18) VEGFC production promoter of present 0.01 invention: Rose oxide (19) Iron oxide As suitable (20) Potassium hydroxide 0.05 (21) Dicalcium glycyrrhizinate 0.01 (22) Pyridoxine hydrochloride 0.01 (23) Ascorbic acid glucoside 0.01 (24) Arbutin 3.0 (25) Saxifraga stolonifera extract 0.1 (26) Water Balance

[0063] Milky Lotion

TABLE-US-00005 Formula (wt %) (1) Butylene glycol 4.0 (2) Propylene glycol 4.0 (3) Carbomer 0.2 (4) Potassium hydroxide 0.2 (5) Behenic acid 0.5 (6) Stearic acid 0.5 (7) Isostearic acid 0.5 (8) Glyceryl stearate 1.0 (9) Glyceryl isostearate 1.0 (10) Behenyl alcohol 0.5 (11) Squalane 5.0 (12) Trioctanoin 3.0 (13) Phenyl trimethicone 2.0 (14) Batyl alcohol 0.5 (15) Dicalcium glycyrrhizinate 0.01 (16) Preservative As suitable (17) Chelating agent As suitable (18) Pigment As suitable (19) VEGFC production promoter of present 0.15 invention: Niaouli oil (20) Purified water Balance

[0064] Cream

TABLE-US-00006 Formula (wt %) (1) Glycerin 10.0 (2) Butylene glycol 5.0 (3) Carbomer 0.1 (4) Potassium hydroxide 0.2 (5) Stearic acid 2.0 (6) Glyceryl stearate 2.0 (7) Glyceryl isostearate 2.0 (8) Vaseline 5.0 (9) Preservative As suitable (10) Antioxidant As suitable (11) VEGFC production promoter of present 0.02 invention: Citronellyl acetate (12) Purified water Balance (13) Chelating agent As suitable (14) Pigment As suitable (15) Stearyl alcohol 2.0 (16) Behenyl alcohol 2.0 (17) Hydrogenated palm oil 2.0 (18) Squalane 10.0 (19) Potassium 4-methoxysalicylate 3.0

[0065] Cream

TABLE-US-00007 Formula (wt %) (1) Glycerin 3.0 (2) Dipropylene glycol 7.0 (3) Polyethylene glycol 3.0 (4) Glyceryl stearate 3.0 (5) Glyceryl isostearate 2.0 (6) Stearyl alcohol 2.0 (7) Behenyl alcohol 2.0 (8) Liquid paraffin 7.0 (9) Cyclomethicone 3.0 (10) Dimethicone 1.0 (11) Octyl methoxycinnamate 0.1 (12) Sodium hyaluronate 0.05 (13) Preservative As suitable (14) Antioxidant As suitable (15) VEGFC production promoter of present 0.02 invention: Rose oxide (16) Purified water Balance (17) Chelating agent As suitable (18) Pigment As suitable

[0066] Gel

TABLE-US-00008 Formula (wt %) (1) Ethyl alcohol 10.0 (2) Glycerin 5.0 (3) Butylene glycol 5.0 (4) Carbomer 0.5 (5) Aminomethyl propanol 0.3 (6) PEG-60 hydrogenated castor oil 0.3 (7) Menthol 0.02 (8) Preservative As suitable (9) Chelating agent As suitable (10) VEGFC production promoter of present 0.01 invention: Citronellyl acetate (11) Purified water Balance

[0067] Aerosol

TABLE-US-00009 Formula (wt %) (1) Glycerin 2.0 (2) Dipropylene glycol 2.0 (3) PEG-60 hydrogenated castor oil 2.0 (4) HPPCD 1.0 (5) Preservative As suitable (6) Chelating agent As suitable (7) Dye As suitable (8) VEGFC production promoter of present 0.1 invention: Niaouli oil (9) Purified water As suitable (10) LPG Balance

[0068] Aerosol

TABLE-US-00010 Formula (wt %) (1) Ethanol 60.0 (2) Methyl lactate 0.1 (3) Sodium lactate 0.1 (4) Tocopherol acetate 0.01 (5) Lactic acid 0.01 (6) Caffeine 0.01 (7) Fennel extract 1.0 (8) Witch hazel extract 1.0 (9) Houttuynia cordata extract 1.0 (10) Dipropylene glycol 1.0 (11) Nitrogen gas 0.0 (12) Polyoxyethylene-polyoxypropylene 1.0 decyl tetradecyl ether (13) Butylene glycol 2.0 (14) Tocopherol 0.05 (15) VEGFC production promoter of present 0.01 invention (16) PEG-60 hydrogenated castor oil 0.1 (17) Water Balance

[0069] Fragrance

TABLE-US-00011 Formula (wt %) (1) Alcohol 75.0 (2) Purified water Balance (3) Dipropylene glycol 5.0 (4) VEGFC production promoter of present 1.0 invention: Citronellyl acetate (5) Antioxidant 8.0 (6) Pigment As suitable (7) Ultraviolet absorber As suitable

[0070] Bath Additive

TABLE-US-00012 Formula (wt %) (1) Sodium sulfate 45.0 (2) Sodium bicarbonate 45.0 (3) Lavender oil 9.0 (4) VEGFC production promoter of present 1.0 invention: Niaouli oil

[0071] Massage Oil

TABLE-US-00013 Formula (wt %) (1) Erythritol 2.0 (2) Caffeine 5.0 (3) Cork tree bark extract 3.0 (4) Glycerin 50.0 (5) Carboxyvinyl polymer 0.4 (6) Polyethylene glycol 400 30.0 (7) Trisodium edetate 0.1 (8) Polyoxylene (10) methylpolysiloxane 2.0 copolymer (9) Squalane 1.0 (10) Sodium hydroxide 0.15 (11) VEGFC production promoter of present 0.01 invention: Rose oxide

[0072] Massage Cream

TABLE-US-00014 Formula (wt %) (1) Solid paraffin 5.0 (2) Beeswax 10.0 (3) Vaseline 15.0 (4) Liquid paraffin 41.0 (5) 1,3-butyleneglycol 4.0 (6) Glycerin monostearate 2.0 (7) POE(20) sorbitan monolaurate 2.0 (8) Borax 0.2 (9) Caffeine 2.0 (10) Preservative As suitable (11) Antioxidant As suitable (12) VEGFC production promoter of present 1.0 invention: Niaouli oil (13) Purified water Balance