COSMETIC COMPOSITION COMPRISING WATER DEPLETED IN 2H AND 18O ISOTOPS

Abstract

The cosmetic composition comprises water as a base and is characterized in that the water contains at most 1100 ppm of .sup.18O and at most 90 ppm of .sup.2H.

Claims

1-31. (canceled)

32: A cosmetic composition comprising water as a base, wherein said water comprises: a) from about 500 ppm to about 1100 ppm of .sup.18O; and b) from about 20 ppm to about 90 ppm of .sup.2H.

33: The cosmetic composition according to claim 32, wherein said water comprises: a) from about 500 ppm to about 1050 ppm of .sup.18O; and b) from about 20 ppm to about 85 ppm of .sup.2H.

34: The cosmetic composition according to claim 33, wherein said water comprises: a) from about 500 ppm to about 1000 ppm of .sup.18O; and b) from about 20 ppm to about 80 ppm of .sup.2H.

35: The cosmetic composition according to claim 32, wherein said water comprises from about 50% to about 90% of the cosmetic composition by weight.

36: The cosmetic composition according to claim 32, wherein said water comprises: a) from about 600 ppm to about 1100 ppm of .sup.18O.

37: The cosmetic composition according to claim 32, wherein said water comprises: b) from about 30 ppm to about 90 ppm of .sup.2H.

38: The cosmetic composition according to claim 32, wherein said composition is essentially free of alcohols.

39: The cosmetic composition according to claim 32, wherein said water is derived from plants comprising one or more substances selected from the group consisting of lipids, polysaccharides, proteins, amino acids and polyphenols.

40: The cosmetic composition according to claim 32, wherein the composition further comprises a non-medicinal active ingredient derived from plant extracts or purified plant-derived agents.

41: The cosmetic composition according to claim 32, wherein said water comprises a weight concentration of residue after evaporation of no more than 5 mg/dm.sup.3.

42: The cosmetic composition according to claim 32, wherein said water comprises a weight concentration of ammonia and ammonium salts of no more than 0.02 mg/dm.sup.3.

43: The cosmetic composition according to claim 32, wherein said water comprises a weight concentration of nitrates of no more than 0.2 mg/dm.sup.3.

44: The cosmetic composition according to claim 32, wherein said water comprises a weight concentration of KMnO.sub.4(O)-reducing substances of no more than 0.08 mg/dm.sup.3.

45: The cosmetic composition according to claim 32, wherein said water has a pH value of from about 5.4 to about 6.6.

46: The cosmetic composition according to claim 32, wherein said water has a specific conductivity at 20 C., S/m, of no more than 5.Math.10.sup.4.

47: The cosmetic composition according to claim 32, wherein said water has a .sup.2H content of no more than 50 ppm.

48: The cosmetic composition according to claim 32, wherein said composition is formulated for use as a topical treatment for ageing, stressed or ailing human skin, hair, nails or lining epithelia.

49: The cosmetic composition according to claim 32, wherein said composition is formulated for use as a topical treatment for attenuating epithelial cell inflammatory responses to biotic and abiotic stresses.

50: The cosmetic composition according to claim 32, wherein said composition is formulated for use as a topical treatment for promoting cellular longevity of skin cells resulting in skin rejuvenation.

51: The cosmetic composition according to claim 32, wherein said composition further comprises a pharmaceutically active drug.

Description

A BRIEF DESCRIPTION OF THE DRAWINGS

[0064] FIG. 1 illustrates the absorbance spectrum of verbascoside plus carboxy methyl cellulose in water depleted from .sup.18O and .sup.2H (De-O,D) after incubation for 30 days;

[0065] FIG. 2 illustrates the dependency of superoxide scavenging activity of plant polyphenol (verbascoside) in % over time; and

[0066] FIG. 3 is a schematic representation of the production of skin equivalents.

DETAILED DESCRIPTION OF THE INVENTION

[0067] The following examples clarify the invention further in more detail. The depleted water was obtained in all examples by physical purification. Extracts of aquatic and terrestrial plant contain organic molecules, such as lipids, polysaccharides, proteins/amino acids, and polyphenols, which are depleted from heavy hydrogen and oxygen isotopes during their synthesis and isotopic fractionation at definite steps of the biosynthesis.

[0068] Rectification of the water was performed in a rectification column in which water vapor generated in the column still is fed directly to the rectification column where an upward flow of vapor interacts with a downward flow of liquid-reflux.

Example 1

[0069] Merystem plant cells containing highly oxydisable active principles such as polyphenols (in this example verbascoside) were homogenized in .sup.18O and .sup.2H depleted (De-O,D) water having a concentration of 99.894 weight-% (and for comparison in distilled natural water). A hydrogel-forming substance, namely carboxyl methyl cellulose was added to the solutions in a concentration of 0.1 weight-%. The final concentration of verbascoside in the mixture was determined to be 100 M by HPLC method using chromatographic standard of verbascoside. This concentration corresponds to 6 mg/100 ml or 0.006 weight-%. The maximum of the absorbance spectrum was obtained at 278 nm and retention time 15.65 min. The mixture was left in darkness at room temperature (25 C.) for thirty days. After this period, the mixture was analyzed for the presence and concentration of verbascoside. The mixture prepared in natural water did not contain any measurable amounts of verbascoside. At the same time, the mixture prepared in .sup.18O and .sup.2H depleted (De-O,D) water retained practically the initial levels of verbascoside measured by HPLC because the absorbance spectrum and the peak at chromatogram corresponding to verbascoside were only slightly changed (>than 5%) as compared to the initial chromatographic data as shown in FIG. 1.

Example 2

[0070] The verbascoside in the mixtures described in Example 1 initially (day 0) possessed superoxide scavenging activity (antioxidant capacity) measured spectrophotometrically by superoxide dismutase dependent reduction of cytochrome c (McCord & Fridovich). In the mixture prepared in natural water of example 1 antioxidant capacity of the polyphenolic molecule verbascoside gradually decayed with the time and was not measurable by day 30 while this capacity remained practically unchanged in the mixture containing De-O,D water, carboxy methyl cellulose and merystem cell homogenate enriched in verbascoside as shown in FIG. 2.

[0071] A scheme of preparation of 30 human skin equivalent is represented in FIG. 3.

[0072] In tables 1 and 2 the date on anti-inflammatory and UV-protective effects of the composition according to the invention towards 3D human skin equivalent is represented.

Example 3

[0073] Convincing data were obtained on the 3D model of human skin equivalents reconstructed in vitro from tiny skin biopsy. The general procedure of the reconstruction is schematically represented in FIG. 3.

[0074] The treatment with .sup.18O and .sup.2H depleted (De-O,D) water: Skin equivalents were cultivated in the appropriate medium (Control) or in the same medium containing 20 ppm or 50 ppm or 80 ppm De-O,D water for 7 days when the skin equivalent was formed.

[0075] Challenge with pro-inflammatory agents: The skin equivalents were exposed to solar simulating UV irradiation containing UVA+UVB (1.0 J/cm2+0.1 J/cm2) in doses corresponding to a daily dose of solar irradiation in summer time. Skin samples were processed and examined 6 h post-irradiation.

[0076] Markers of inflammatory response from human skin cells: The gene expression of TNFalpha, IL-6, and IL-8 as markers of inflammatory response of human skin equivalents was measured by real-time PCR method using molecular primers constructed for each gene. The gene expression of cycloxygenase-2 (COX-2) as a measure of the risk of carcinogenesis was measured by the same approach. Experiments were carried out on three skin cell cultures derived from three different donors. Each measurement was repeated 3-4 times and results were statistically evaluated using the Student's test.

Results:

[0077] The presence of De-O,D water at 80 ppm completely inhibited inflammatory response from human skin cells from TNFalpha and IL-6 induced by solar simulating UVA+UVB irradiation. The presence of De-O,D water at 50 ppm was less effective in the inhibition of TNFalpha and IL-6 expression whereas De-O,D water at 20 ppm exerted inhibitory efficacy similar to that of 80 ppm De-O,D water (Table 1). [0078] The presence of .sup.18O and .sup.2H depleted (De-O,D) water at 80 ppm partially but substantially {by 30-70%) inhibited the induction of IL-8 and COX-2 gene expression by UV irradiation. At the same time, .sup.18O and .sup.2H depleted (De-O,D) water at 50 ppm and 20 ppm was much less effective (Table 1). [0079] Anti-inflammatory potential of .sup.18O and .sup.2H depleted (De-O,D) water against UV irradiation is extremely high and comparable with the effects of the most effective anti-inflammatory drugs.

Example 4

[0080] 1. Convincing data were obtained on the 3D model of human skin equivalents reconstructed in vitro from tiny skin biopsy. The general procedure of the reconstruction is schematically presented in FIG. 3.

[0081] The treatment with .sup.18O and .sup.2H depleted (De-O,D) water: Skin equivalents were cultivated in the appropriate medium (Control) or in the same medium containing 20 ppm or 50 ppm or 80 ppm De-O,D water for 7 days when the skin equivalent was formed.

[0082] Challenge with bacterial inflammation-inducing agent: The skin equivalents were treated with 1 microg/ml of bacterial lipopolysaccharide (LPS from E. coli) to induce inflammatory response of human skin cells to bacterial component.

2. The De-O,D water at 80 ppm completely inhibited gene expression of IL-6 and COX-2 induced by LPS while increased the expression of TNFalpha and IL-8 induced by bacterial lipopolysaccharide as shown in table 2.
3. The De-O,D water at 50 ppm was more efficient in the inhibition of TNFalpha, IL-6 and COX-2 while less efficient in the induction of IL-8 as shown in table 2.
4. The De-O,D water at 20 ppm has the greatest effect on COX-2 inhibition and IL-8 induction.

[0083] On the grounds of these data, .sup.18O and .sup.2H depleted (De-O,D) water with 80-20 ppm range from may be useful in the cosmetic and dermatologic preparations (gels, creams, lotions, etc.) with UV-protective, anti-inflammatory, and anti-bacterial claims.

Determination of Deuterium in the Finished Product

[0084] Deuterium content was determined by means of time-based cavity ring-down spectroscopy (CRDS) using a Picarro L2130i/L2140i water isotopic composition analyzer (for D and .sup.18O) or by means of mass spectroscopy using a Thermo Scientific (Finnigan) series Delta isotope mass spectrometers with an H-Device (Thermo Scientific) peripheral device.

[0085] The residual content of deuterium in the final product is expressed in p.p.m. or as a deviation from the International Standard VSMOW (), expressed in ppm ().

TABLE-US-00001 TABLE 1 De-O,D water against UV-induced inflammation Cytokine/System Gene expression Significance TNFalpha Skin equivalent (SE, control) 1.02 0.21 SE + UV 2.15 0.45* p < 0.01 vs control SE + UV + 80 ppmDe-O,D 1.00 0.16** p < 0.01 vs UV SE + UV + 50 ppmDe-O,D 1.18 0.17** p < 0.05 vs UV SE + UV + 20 ppmDe-O,D 0.95 0.13** p < 0.01 vs UV IL-6 SE (control) 1.00 0.12 SE + UV 13.29 2.69* p < 0.01 vs control SE + UV + 80 ppmDe-O,D 1.13 0.15** p < 0.01 vs UV SE + UV + 50 ppmDe-O,D 2.03 0.21** p < 0.05 vs UV SE + UV + 20 ppmDe-O,D 0.90 0.12** p < 0.01 vs UV COX-2 SE (control) 1.00 0.12 SE + UV 3.00 0.59* p < 0.01 vs control SE + UV + 80 ppmDe-O,D 1.35 0.13** p < 0.01 vs UV SE + UV + 50 ppmDe-O,D 1.55 0.13** p < 0.05 vs UV SE + UV + 20 ppmDe-O,D 1.83 0.22** p < 0.05 vs UV IL-8 SE (control) 1.00 0.00 SE + UV 4.23 0.84* p < 0.01 vs control SE + UV + 80 ppmDe-O,D 125 0.13** p < 0.01 vs UV SE + UV + 50 ppmDe-O,D 2.68 0.35** p < 0.05 vs UV SE + UV + 20 ppmDe-O,D 2.75 0.31** p < 0.05 vs UV

TABLE-US-00002 TABLE 2 De-O,D water and inflammatory/anti-bacterial responses induced by bacterial LPS Cytokine/System Gene expression Significance TNFalpha SE (control) 1.01 0.03 SE + UV 1.23 0.15* p < 0.05 vs control SE + UV + 80 ppmDe-O,D 2.45 0.51** p < 0.01 vs LPS SE + UV + 50 ppmDe-O,D 2.20 0.52** p < 0.01 vs LPS SE + UV + 20 ppmDe-O,D 2.53 0.15** p < 0.01 vs LPS IL-6 SE (control) 1.00 0.12 SE + UV 1.67 0.23* p < 0.01 vs control SE + UV + 80 ppmDe-O,D 1.00 0.18** p < 0.01 vs LPS SE + UV + 50 ppmDe-O,D 0.80 0.18** p < 0.01 vs LPS SE + UV + 20 ppmDe-O,D 0.98 0.05** p < 0.01 vs LPS COX-2 SE (control) 1.00 0.12 SE + UV 0.88 0.10 p > 0.05 vs control SE + UV + 80 ppmDe-O,D 0.70 0.12 p > 0.05 vs LPS SE + UV + 50 ppmDe-O,D 0.55 0.13** p < 0.05 vs LPS SE + UV + 20 ppmDe-O,D 0.45 0.06** p < 0.05 vs LPS IL-8 SE (control) 1.00 0.00 SE + UV 1.20 016 p > 0.05 vs control SE + UV + 80 ppmDe-O,D 4.70 1.10** p < 0.01 vs LPS SE + UV + 50 ppmDe-O,D 4.00 0.87** p < 0.01 vs LPS SE + UV + 20 ppmDe-O,D 6.13 0.13** p < 0.01 vs LPS

Example 5 (Body Lotion)

[0086] 59 weight-% of Citrus lemon fruit water with a content of 80 ppm .sup.2H and 1000 ppm of .sup.18O [0087] 7.5 weight-% Persea gratissima oil [0088] 5.7 weight-% Aleurites Moluccana seed oil [0089] 4.6 weight-% glycerin [0090] 23.2 weight-% mixture of various ingredients used for body lotions.

Example 6 (Creme)

[0091] 68 weight-% De-O, D water having a content of 50 ppm .sup.2H and 850 ppm of .sup.18O [0092] 9 weight-% cetearyl isononanoate [0093] 5 weight-% stearic acid [0094] 4 weight-% caprylic triglyceride [0095] 14 weight-% mixture of various ingredients used for cremes

Example 7 (Facial Mask)

[0096] 55 weight-% De-O, D water having a content of 50 ppm .sup.2H and 950 ppm of .sup.18O [0097] 20 weight-% glycerin [0098] 15 weight-% propylene glycol [0099] 5 weight-% pullulan [0100] 5 weight-% mixture of various ingredients used for facial masks

Example 8 (Facial Spray)

[0101] 98.5 weight-% De-O, D water having a content of 45 ppm .sup.2H and 930 ppm of .sup.18O [0102] 0.95 weight-% phenoxyethanol [0103] 0.2 weight-% polysorbate 20 [0104] 0.35 weight-% mixture of various ingredients used for facial sprays.

Example 9 (Hand Care)

[0105] 86.2 weight-% De-O, D water having a content of 75 ppm .sup.2H and 990 ppm of .sup.18O [0106] 5.5 weight-% caprylic triglyceride [0107] 2.5 weight-% glyceryl stearate citrate [0108] 1.5 weight-% cetearyl alcohol [0109] 4.3 weight-% mixture of various ingredients used for hand care

Example 10 (Oral Gel)

[0110] 77.4 weight-% De-O, D water having a content of 48 ppm .sup.2H and 945 ppm of .sup.18O [0111] 10.5 weight-% lactobacillus/papaya fruit ferment extract in an aqueous solution of De-O, [0112] D water having a content of 80 ppm .sup.2H and 1000 ppm of .sup.18O [0113] 8 weight-% propylene glycol [0114] 3.15 weight-% hydroxypropylmethylcellulose (HPMC) [0115] 0.95 weight-% mixture of various ingredients used for oral gels

Example 11 (Serum)

[0116] 75.7 weight-% De-O, D water having a content of 20 ppm .sup.2H and 750 ppm of .sup.18O [0117] 6 weight-% propylene glycol [0118] 3 weight-% pentylene glycol [0119] 2.6 weight-% haxapeptide-11 [0120] 2.46 weight-% methylpropanediol [0121] 2.0 weight-% betaine [0122] 8.44 weight-% mixture of various ingredients used for serums

Example 12 (Skin Aerosol Spray)

[0123] 97.99998706 weight-% De-O, D water having a content of 47 ppm .sup.2H and 945 ppm of .sup.18O [0124] 2 weight-% nitrogen [0125] 0.00001294 silver chloride

Example 13 (Vaginal Gel)

[0126] 85.4 weight-% De-O, D water having a content of 55 ppm .sup.2H and 950 ppm of .sup.18O [0127] 10 weight-% lactobacillus/papaya fruit ferment extract in an aqueous solution of De-O, D water having a content of 80 ppm .sup.2H and 1000 ppm of .sup.18O [0128] 3.15 weight-% hydroxypropylmethylcellulose (HPMC) [0129] 0.6 weight-% sodium lactate [0130] 0.85 weight-% mixture of various ingredients used for vaginal gels.

Example 14 (Veterinary Spray)

[0131] 95.98 weight-% De-O, D water having a content of 80 ppm .sup.2H and 1000 ppm of 180 [0132] 1.98 weight-% PEG-40 hydrogenated castor oil [0133] 1.0 weight-% menthol [0134] 1.04 weight-% mixture of various ingredients used for veterinary sprays

[0135] Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the scope of the appended claims.

[0136] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.