TOPICAL SKIN LIGHTENING ADDITIVE AND COMPOSITION WITH AMINO ACIDS AND NICOTINAMIDE COMPOUNDS

Abstract

Skin lightening additives and personal care compositions containing amino acids and nicotinamide compounds. The compositions are useful for attaining even skin color and reducing pigmentation, age spots and discoloration.

Claims

1. A topical skin lightening composition comprising: a. skin lightening additive comprising by weight of the composition: i. from about 0.001 to about 2% of cystine; ii. from about 0.01 to about 10% of a glutamate source selected from the group consisting of glutamine, glutamic acid, pyroglutamic acid, and mixtures thereof; iii. from about 0.01 to about 10% of glycine; and iv. from about 0.05 to about 10% of a nicotinamide compound selected from the group consisting of Formula I, Formula IL Formula III, and mixtures thereof: ##STR00002## wherein R is CONH.sub.2 or CH.sub.2OH or COOR2; R.sub.1 is C1 to C5 straight or branched alkyl group or a furanosyl group or a deoxyfuranosyl group or a pyranosyl group or a deoxypyranosyl group; R.sub.2 is C1-C24 straight or branched alkyl or alkenyl group; and b. a cosmetically acceptable carrier; c. wherein the pH of the composition is in the range of from about 3.5 to about 8.5

2. The composition of claim 1 wherein nicotinamide compound is nicotinamide riboside.

3. The composition of claim 1, wherein the glutamate source is pyroglutamic acid or salt thereof.

4. The composition of claim 1 wherein nicotinamide compound is N-methyl nicotinamide.

5. The composition of claim 1 wherein nicotinamide compound is nicotinamide.

6. The composition of claim 1 wherein nicotinamide compound is isonicotinamide.

7. The composition of claim 1 wherein nicotinamide compound is nicotinyl myristate.

8. The composition of claim 1 wherein the composition is in the form of a water-in-oil emulsion comprising cystine in an aqueous phase, wherein 90% of the water droplets have a diameter within the size range of from 100 nm to 20 microns.

9. The composition of claim 1 comprising from 0 to at most 0.1% of selenium source

10. The composition of claim 1 wherein the composition is a leave-on non-solid skin cosmetic composition.

11. The composition of claim 1 wherein the composition is a vanishing cream.

12. The composition of claim 1 wherein the composition further comprises 4-alkyl resorcinol.

13. The composition of claim 1 wherein the composition further comprises 12-hydroxystearic acid.

14. Method of improving the appearance of skin comprising applying to the skin the composition of claim 1.

15. Method of attaining even skin color and reducing pigmentation, age spots and discoloration, comprising applying to the skin the composition of claim 1.

Description

EXAMPLES

Experimental Methods

[0099] Powders of the following amino acids (glutamine and glycine) were purchased from Sigma, and individual stock solutions of each amino acid prepared by reconstitution of the powder in water (pH 7). Cystine (Sigma) stock solutions were generated in 0.5M sodium hydroxide (pH 12) as cystine is poorly soluble in neutral solutions. To generate the mixture henceforth described as GAP, appropriate volumes of three amino acids (glutamine, cystine, and glycine) were combined in Hank's Balanced Salt Solution (Sigma) such that the glutamine, cystine, and glycine were in a 0.5:1:1 weight ratio. The specific concentrations used in each assay are referenced below. Powders of cystine esters, including ditert-butyl L-cystinate dihydrochloride (DTBC) and diethyl L-cystinate dihydrochloride (DEC) were purchased from Bachem; L-Cystine dimethyl ester dihydrochloride (CDME) was from Sigma. The cystine esters were prepared in water and stock solutions were diluted in Hank's Balanced Salt Solution (HBSS). The specific concentrations used in each assay are referenced below. Additional test reagents: nicotinamide (Sigma-Aldrich), niacin (Himedia), nicotinamide riboside (Chromadex), isonicotinamide (Sigma-Aldrich), picolinamide (Sigma-Aldrich), n-methyl-nicotinamide (Sigma-Aldrich). Nicotinamide and derivatives/related compounds were solubilized in water.

Melanin Content Assay

[0100] Human primary melanocytes (Cascade Biologics, now ThermoFisher Scientific) were grown in melanocytes growth medium (MGM) with human melanocyte growth supplement (both media components from ThermoFisher Scientific). Melanocytes were seeded at 510.sup.4 cells/well in 24 well -plates and incubated in a humidified incubator at 37 C. with 5% CO.sub.2 atmosphere. After 24 hours of incubation, cell cultures were treated with the GAP amino acids or additional test reagents above-described, alone or in combination, as described in examples below. The vehicle control consisted of cells treated with the vehicle (0.002 mM NaOH) alone. After treatment, cells were again incubated for 72 hours in a humidified incubator at 37 C. with 5% CO2 atmosphere. At the end of the 72 hour incubation, the culture medium was replaced with 120 uL/well of melanin content assay reagent (10% DMSO in 1M sodium hydroxide) and incubated for 1 hour at 60 C. in a shaker incubator. The supernatant was then transferred to a 384 well plate. The absorbance was measured by spectrophotometric analysis at 405 nm with a micro plate reader (GEnios Pro, Tecan).

The spectrophotometric optical density at 405 nm for untreated cells (no active) was considered as 100% melanin content. Relative melanin content for treated samples was calculated with normalization based on the untreated (media/vehicle) control.

Tyrosinase Activity Assay

[0101] Human primary melanocytes (Cascade Biologics, now ThermoFisher Scientific) were grown in melanocytes growth medium (MGM) with human melanocyte growth supplement (both media components from ThermoFisher Scientific). Melanocytes were seeded at 110.sup.4 cells/well in a 96 wells plate and cultures left undisturbed for 24 hours in a 5% CO.sub.2 humidifed incubator at 37 C. At 24 hrs post-seeding, cells were treated with GAP amino acids or additional test reagents above-described, alone or in combination, as described in examples below and left undisturbed for another 72 hours. Subsequently, cell viability was assessed and the cells were then lysed and progressed for determination of in situ tyrosinase activity. Cultures were rinsed twice with 1 phosphate-buffered saline and lysed with 40 uL of 0.5% TritonX 100 for 1 hr on an ice bed. In situ tyrosinase activity was visualized by addition of 60 l of 50 mM potassium phosphate buffer (pH 6.8) containing 2 mM DOPA (3,4 dihydroxy phenylalanine) and 4 mM MBTH reagent (3-Methyl-2-Benzothiazolinone hydrazone hydrochloride) for 1 hour at 37 C. The reaction was stopped by the addition of 100 uL of ice-cold 10% tricholoracetic acid and then centrifuged at approximately 300 g for 10 minutes at 4 C. The soluble supernatant was separated from the pellet and the optical density read in a TECAN plate-reader (540 nm filter). The spectrophotometric optical density at 540 nm for untreated cells (media/vehicle) is considered as 100% tyrosinase activity. Tyrosinase activity is expressed after correction for cell numbers (activity/viability) and represented as % control.

Glutathione Assay

[0102] Human Keratinocytes (Promocell, Heidelberg, Germany) were maintained in EpiLife medium containing 60 uM calcium chloride and 1% Human Keratinocyte Growth Supplement (both media components from ThermoFisher Scientific) in a humidified incubator with an atmosphere containing 5% CO.sub.2 at 37 C. The medium was refreshed every 2-3 days. Subsequently, cells were trypsinized at 90% confluency and seeded at a density of 110.sup.4 per well into 96-well plates with white wall and clear bottom. One or two days after seeding, cell medium was replaced with Hank's Balanced Salt Solution (HBSS), and keratinocytes were pre-treated with GAP (or 0.12-0.240 mM sodium hydroxide vehicle control, pH 7.5) alone or in combination with additional test reagents described above or L-cystine esters alone for 1-2 hours. The concentration of all amino acids constituting the GAP mixture used in this assay are designated in the data tables, and fully described in the abbreviations list below.

[0103] After one or two hours of pre-treatment with GAP or additional test reagents, alone or in combination, keratinocytes were then challenged with 25 uM menadione (Sigma-Aldrich), a known inducer of oxidative stress. After 18-20 hours, cells were harvested and analyzed for total glutathione levels using a commercially available kit (GSH-GSSG GLo Assay, Promega), a luminescence-based system for the detection and quantification of total glutathione in cultured cells. In summary, after cell treatment, the media was removed and replaced with a cell lysis reagent for 5 minutes at 20-25 C.; note, all remaining steps were also conducted at 20-25 C. Next, a luciferase generation reagent was added to each well and incubated for 30 minutes. Following this step, a luciferin detection reagent was added to each well and plates were incubated for 15 minutes. Finally, plates were then read for luminescence in a FlexStation 3 plate reader (Molecular Devices). Total glutathione levels were calculated after interpolation of glutathione concentrations from a standard curve. Experimental conditions were typically conducted in at least triplicate determinations. Statistical significance was calculated using 1-way ANOVA. Percent protection was calculated by using the calculated glutathione levels from each experiment in the following equation:

[00001]$\frac{\left(\mathrm{Test}.Math..Math.\mathrm{Sample}\right)-\left(\mathrm{vehiclecontrol}+\mathrm{menadione}\right)}{\mathrm{vehiclecontrol}}100.Math.\%=\%.Math..Math.\mathrm{protection}$

Reactive Oxygen Species (ROS) Assay

[0104] Human Keratinocytes (Promocell, Heidelberg, Germany) were maintained in EpiLife keratinocyte medium containing 60 uM calcium chloride and 1% Human Keratinocyte Growth Supplement (HKGS) in a humidified incubator with an atmosphere containing 5% CO2 at 37 C. The medium was refreshed every other day. Subsequently, cells were trypsinized at 90% confluency and seeded at a density of 1-210.sup.4 per well into 96-well plates with black wall and clear bottom. On the second day after seeding, cell medium was replaced with Hank's Balanced Salt Solution (HBSS) with or without menadione (10-25 uM), GAP, GAP vehicle (0.2 mM NaOH, pH 7.5), above-described additional test reagents, or combinations of GAP with additional test reagents or L-cystine esters alone, and incubated for additional 18 hours. For ROS detection, CelIROX green (Life Tech, Thermo Fisher Scientific, Rockford, USA) reagent was added into each well to a final concentration of 15-25 uM and incubated for additional 2-4 hours at 37 C. in a humidified incubator. The ROS fluorescence was detected using a Tecan microplate reader (Excitation/Emission=490/525). Relative ROS production was calculated with normalization based on the vehicle control treatment which was set to 100%. Where indicated, percent protection was calculated as a comparison to menadione alone treatment, using the measured relative fluorescent units in the following equation:

[00002]$\frac{\left(\mathrm{Vehiclecontrol}+\mathrm{Menadione}\right)-\left(\mathrm{Testsample}\right)}{\left(\mathrm{vehiclecontrol}+\mathrm{menadione}\right)}100.Math.\%=\%.Math..Math.\mathrm{protection}$

[0105] ROS levels were evaluated together with cell viability because it is possible to get a false negative for ROS if treatment is so toxic, that at the time of assay, cells are dead (and there are no longer any detectable ROS). Decrease in ROS levels is meaningful if cells are still viable.

Cell Viability Assay

[0106] Human Keratinocytes (Promocell, Heidelberg, Germany) were maintained in EpiLife keratinocyte medium containing 60 uM calcium chloride and 1% Human Keratinocyte Growth Supplement (HKGS). The medium was refreshed every other day. Subsequently, cells were trypsinized at 90% confluency and seeded at a density of 210.sup.4 per well into 96-well transparent tissue culture plates. On the second day after seeding, cell medium was replaced with treatments as described above in ROS assay section. After 18 hours, Cell Culture Kit-8 (CCK-8) reagent (Dojindo, Kumamoto, Japan) was diluted 1:10 in HBSS and incubated with cells for 4 h at 37 C. in a humidified incubator. Optical absorption at 450 nm was measured using a Tecan Safire2 plate reader. Relative viability was calculated with normalization based on the vehicle control treatment, which was set to 100%.

[0107] Where indicated, percent protection was calculated as a comparison to menadione alone treatment, using the measured relative absorbance units in the following equation:

[00003]$\frac{\mathrm{TestSample}-\left(\mathrm{vehicle}.Math..Math.\mathrm{control}-\mathrm{menadione}\right)}{\left(\mathrm{vehiclecontrol}-\text{?}\right)}100.Math.\%=\%.Math..Math.\mathrm{protection}$$\text{?}.Math.\text{indicates text missing or illegible when filed}$

[0108] Abbreviations in the Examples are as follows:

[0109] mM=millimolar

[0110] uM=micromolar

[0111] uL=micro liter

[0112] GAP=glutamine (32 uM), cystine (40 uM), glycine (129 uM) except where otherwise denoted below.

[0113] GAP 10=8 uM sodium pyroglutamic acid, 10 uM cystine, 32 uM glycine

[0114] GAP 20=16 uM sodium pyroglutamic acid, 20 uM cystine, 64 uM glycine

[0115] GAP 60=48 uM sodium pyroglutamic acid, 60 uM cystine, 192 uM glycine

[0116] HR=4-hexylresorcinol

[0117] DEC=Diethyl L-cystinate dihydrochloride

[0118] CDME=L-Cystine dimethyl ester dihydrochloride

[0119] DTBC=Ditert-butyl L-cystinate dihydrochloride

Example 1

[0120] Example 1 investigated the skin lightening potential of the inventive compositions. The results that were obtained are summarized in Table 1A through 1E.

TABLE-US-00001 TABLE 1A Melanin synthesis inhibition Sample Mean of % Inhibition A Media Control 0 B Vehicle Control 0 Test samples C HR (10 uM) 35.sup.1 D GAP 1 E GAP.sup.4 14.sup.1 F Nicotinamide (10 mM) 10.sup.1 G Nicotinamide (5 mM) 9.sup.1 1 GAP + Nicotinamide (10 mM) 27.sup.1, 2, 3 2 GAP + Nicotinamide (5 mM) 14.sup.1, 2, 3 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control .sup.2p < 0.05 compared to GAP (sample D) .sup.3p < 0.05 compared to Nicotinamide alone (at equivalent dose) .sup.4amino acid concentrations: glutamine (954 uM), cystine (1194 uM), glycine (129 uM)

TABLE-US-00002 TABLE 1B Tyrosinase inhibition Sample Mean of % Inhibition A Media Control 0 B Vehicle Control 0 Test Samples C HR (10 uM) 67.sup.1 D GAP 9.sup.1 E Nicotinamide (10 mM) 24.sup.1 F Nicotinamide (5 mM) 16.sup.1 1 GAP + Nicotinamide (10 mM) 42.sup.1, 2, 3 2 GAP + Nicotinamide (5 mM) 20.sup.1 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control .sup.2p < 0.05 compared to GAP .sup.3p < 0.05 compared to Nicotinamide alone (at equivalent dose)

TABLE-US-00003 TABLE 1C Melanin synthesis inhibition Mean of % Sample Inhibition A Media Control 0 B Vehicle Control 0 Test Samples C HR (10 M) 26.sup.1 D N-methyl Nicotinamide (10 mM) 7.sup.1 E N-methyl Nicotinamide (5 mM) 0 F GAP 2 1 GAP + N-methyl Nicotinamide (10 mM) 12.sup.1,2,3 2 GAP + N-methyl Nicotinamide (5 mM) 4.sup.1 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control .sup.2p < 0.05 compared to GAP .sup.3p < 0.05 compared to N-methyl Nicotinamide alone (at equivalent dose)

TABLE-US-00004 TABLE 1D Melanin synthesis inhibition Mean of % Sample Inhibition A Media Control 0 B Vehicle Control 0 Test Samples C HR (10 uM) 32.sup.1 D GAP 1 E Isonicotinamide (10 mM) 20.sup.1 F Isonicotinamide (5 mM) 4.sup.1 1 GAP + Isonicotinamide (10 mM) 25.sup.1,2 2 GAP + Isonicotinamide (5 mM) 11.sup.1,2,3 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control .sup.2p < 0.05 compared to GAP .sup.3p < 0.05 compared to iso-nicotinamide alone (at equivalent dose)

TABLE-US-00005 TABLE 1E Melanin synthesis inhibition Mean of % Sample Inhibition A Media Control 0 B Vehicle Control 0 Test Samples C HR (10 uM) 28.sup.1 D GAP 1 E Nicotinamide Riboside (2.5 uM) 9.sup.1 F Nicotinamide Riboside (1 uM) 4.sup.1 1 GAP + Nicotinamide Riboside (2.5 uM) 9.sup.1,2 2 GAP + Nicotinamide Riboside (1 uM) 8.sup.1,2,3 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control .sup.2p < 0.05 compared to GAP .sup.3p < 0.05 compared to nicotinamide riboside alone (at equivalent dose)

[0121] In Tables 1A through 1E, samples marked by alphabetical characters were outside the scope of the invention. Numerically marked samples were within the scope of the invention. Sample C in Tables 1A through 1E was a positive control as it employed 4-hexyl resorcinol, a known highly effective skin lightening agent. Overall, it can be seen from the results in Tables 1A 1E that combination of nicotinamide compounds within the scope of the invention with GAP amino acids resulted in synergistic improvement (inhibition of melanin synthesis and tyrosinase inhibition) compared to the results achieved with either the nicotinamide compound or GAP alone.

Comparative Example 2

[0122] Example 2 investigated the skin lightening potential of combinations of GAP amino acids with either picolinamide (structurally similar molecule to nicotinamide) or niacin (nicotinic acid)both outside the scope of the invention. The results that were obtained are summarized in Tables 2A and 2B.

TABLE-US-00006 TABLE 2 Melanin synthesis inhibition Sample Mean of % Inhibition A Media Control 0 B Vehicle Control 0 Test Samples C HR (10 uM) 36.sup.1 D GAP 1 E Picolinamide (5 mM) 6 F Picolinamide (10 mM) 32.sup.1 G GAP + Picolinamide (10 mM) 31.sup.1,2 H GAP + Picolinamide (5 mM) 16.sup.1,2 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control .sup.2p < 0.05 compared to GAP

TABLE-US-00007 TABLE 2B Melanin Synthesis inhibition Mean of % Sample Inhibition A Media Control 0 B Vehicle Control 0 Test Sammples C HR (10 uM) 32.sup.1 D GAP 1 E Niacin (5 mM) 7 F Niacin (10 mM) 14.sup.1 G GAP + Niacin (10 mM) 5 H GAP + Niacin (5 mM) 5 Statistically significant improvement: .sup.1p < 0.05 compared to media and vehicle control

[0123] It can be seen from the results in Tables 2A and 2B that combination of GAP amino acids with either picolinamide or niacin (both outside the scope of the invention) did not result in synergistic improvement (inhibition of melanin synthesis) compared to the results achieved with either the nicotinamide compound or GAP alone.

Example 3

[0124] Example 3 evaluated amino acids constituting GSH building blocks for potentiating intracellular GSH production. Results that were obtained are summarized in Tables 3A In Tables 3B through 3D the efficacy of various cystine esters was also tested.

TABLE-US-00008 TABLE 3A Glutathione Synthesis % Protection (improvement vs. Mean (uM vehicle control + Sample glutathione) menadione) A Vehicle Control 1.75 B Vehicle Control + 0.07.sup.1 0.sup.1 menadione Test Samples C GAP 10 + menadione 1.34.sup.1,2 72.sup.1,2 D GAP 20 + menadione 4.61.sup.2 259.sup.2 Statistically significant decrease: .sup.1p < 0.05 compared to HR + GAP10 + menadione Statistically significant improvement: .sup.2p < 0.05 compared to vehicle control + menadione

TABLE-US-00009 TABLE 3B Glutathione Syntheiss % Protection (improvement vs. Mean (m vehicle control + Sample glutathione) menadione) A Vehicle control 2.73 B Vehicle control + Menadione 0.18.sup.1 0 C DEC (40 uM) + Menadione 0.54.sup.1,2 13.sup.1,2 D DEC (80 uM) + Menadione 2.72.sup.2 93.sup.2 Statistically significant decrease: .sup.1p < 0.05 compared to vehicle control Statistically significant improvement: .sup.2p < 0.05 compared to vehicle control + menadione

TABLE-US-00010 TABLE 3C Glutathione Synthesis % Protection (improvement vs. Mean (m vehicle control + Sample glutathione) menadione) A Vehicle control 2.73 B Vehicle control + Menadione 0.18.sup.1 0 C DTBC (40 uM) + Menadione 0.17.sup.1 0.sup.1 D DTBC (80 uM) + Menadione 0.19.sup.1 1.sup.1 Statistically significant decrease: .sup.1p < 0.05 compared to vehicle control

TABLE-US-00011 TABLE 3D Glutathione Synthesis % Protection (improvement vs. Mean (m vehicle control + Sample glutathione) menadione) A Vehicle control 2.96 B Vehicle control + Menadione 0.18.sup.1 0 C CDME (80 uM) + Menadione 1.09.sup.1,2 31.sup.1,2 Statistically significant decrease: .sup.1p < 0.05 compared to vehicle control Statistically significant improvement: .sup.2p < 0.05 compared to vehicle control + menadione

Example 4

[0125] Example 4 evaluated amino acids constituting GSH building blocks for improved reduction in oxygen species. Results that were obtained are summarized in Table 4A In Tables 4B through 4D the efficacy of various cystine esters was also tested.

TABLE-US-00012 TABLE 4A Reduction in Oxygen Species Mean ROS % protection % Protection production (improvement Cell (improvement (% of vs vehicle viability vs vehicle vehicle control + (% of control + Sample control) menadione) control) menadione) A Vehicle 100.0 100.0 Control B Vehicle 504.8.sup.1 54.8.sup.1 Control + Menadione Test Samples C Menadione 171.5.sup.1,2 66.02.sup.1,2 92.7.sup.1,2 69.11.sup.1,2 GAP 60 Statistically significant difference: .sup.1p < 0.05 compared to vehicle control Statistically significant difference: .sup.2p < 0.05 compared to vehicle control + menadione

TABLE-US-00013 TABLE 4B Reduction in Reactive Oxygen Species ROS % Protection % Protection inhibition (improvement Cell (improvement (% of vs. vehicle viability vs vehicle vehicle control + (% of control + Sample control) menadione) control) menadione) Vehicle control 100 100 Vehicle control + 375.5.sup.1 73.7.sup.1 Menadione DEC (20 uM) + 232.8.sup.1,2 38.0.sup.1,2 109.8.sup.2 48.9.sup.2 Menadione DEC (40 uM) + 211.9.sup.1,2 43.6.sup.1,2 127.3.sup.1,2 72.7.sup.1,2 Menadione DEC (80 uM) + 233.8.sup.1,2 37.7.sup.1,2 131.5.sup.1,2 78.3.sup.1,2 Menadione DEC (160 uM) + 253.8.sup.1,2 32.4.sup.1,2 122.3.sup.1,2 65.9.sup.1,2 Menadione Statistically significant difference: .sup.1p < 0.05 compared to vehicle control Statistically significant difference: .sup.2p < 0.05 compared to vehicle control + menadione

TABLE-US-00014 TABLE 40 Reduction in Oxygen Species DTBC (160 uM)+ Menadione 221.1.sup.1, 2 ROS (improvement viability (improvement inhibition vs. vehicle (% of vs. vehicle (% of vehicle control + vehicle control + Sample control) menadione) control) menadione) Vehicle control 100 100 Vehicle control + 375.5.sup.1 73.7.sup.1 Menadione DTBC (20 uM) + 367.0.sup.1 2.2 78.1.sup.1 6.0 .sup.1 Menadione DTBC (40 uM) + 323.7.sup.1, 2 13.8.sup.1, 2 75.9.sup.1 3.0.sup.1 Menadione DTBC (80 uM) + 268.7.sup.1, 2 28.4.sup.1, 2 73.6.sup.1 0.1.sup.1 Menadione Statistically significant difference: .sup.1p < 0.05 compared to vehicle control Statistically significant difference: .sup.2p < 0.05 compared to vehicle control + menadione indicates data missing or illegible when filed

TABLE-US-00015 TABLE 4D Reduction in Oxygen Species ROS % Protection Cell % protection inhibition (improvement viability (improvement (% of vs. vehicle (% of vs. vehicle vehicle control + vehicle control + Sample control) menadione) control) menadione) Vehicle control 100 100 Vehicle control + 131.8.sup.1 69.8.sup.1 Menadione CDME (20 uM) + 107.8.sup.2 18.2.sup.2 90.9.sup.1,2 30.2.sup.1,2 Menadione CDME (40 uM) + 111.3.sup.1,2 15.5.sup.1,2 93.1.sup.1,2 33.4.sup.1,2 Menadione CDME (80 uM) + 114.3.sup.1,2 13.3.sup.1,2 87.0.sup.1,2 24.6.sup.1,2 Menadione CDME (160 uM) + 97.9.sup.2 25.7.sup.2 105.9.sup.1,2 51.7.sup.1,2 Menadione Statistically significant difference: .sup.1p < 0.05 compared to vehicle control Statistically significant difference: .sup.2p < 0.05 compared to vehicle control + menadione

Example 5

[0126] Personal care formulations according to the present invention are illustrated in the Tables below. All numbers in the Tables represent weight % in the composition.

TABLE-US-00016 TABLE I Oil-in-water formulations, lotions, and creams OW-1 OW-2 OW-3 OW-4 OW-5 Water To 100 To 100 To 100 To 100 To 100 Glycerine 0-40 1-40 1-5 1-10 1-40 Propylene glycol 0-5 0-5 Butylene glycol 0-5 0-5 0-5 Carbomer 0-2 0.03-1 Ammonium 0-1 0.03-1 0.01-1 Acryloyl dimethyl taurate/VP copolymer Styrene/ 0-1 0.01-1 Acrylates copolymer Xanthan Gum 0-1 0.01-1 EDTA 0.01-0.01 0.01-0.01 0.01-1 0.01-1 0.01-1 Preservative 0.02-2 0.02-2 0.02-2 0.02-2 0.02-2 Titanium oxide 0-10 0.01-10 0.01-10 0.01-10 0.01-10 Colorant/ 0-5 0-5 0-5 0-5 0-5 Pigment Triethanol 0-3 0.01-3 0.01-3 0.01-3 0.01-3 amine/Sodium Hydroxide/ potassium Hydroxide Stearic acid 0-5 0.01-5 0.01-5 0.01-5 0.01-5 Isopropyl 0-10 0.01-10 Myristate Capric/Caprylic 0-10 0.01-10 Triglyceride C12-C15 alkyl 0-10 0.01-10 benzoate Mineral oil 0-10 0.01-10 Glyceryl stearate 0-5 0.01-5 Steareth-2 0-5 0.01-5 0.01-5 Steareth-21 0-5 0.01-5 Peg100 Stearate 0-5 0.01-2 0.01-5 Potassium Cetyl 0-5 0.01-2 Phosphate Tween20 0-5 0.01-5 Cetyl alcohol 0-4 0.01-4 0.01-4 Dicaprylyl 0-5 0.01-5 carbonate Ethyl hexyl 0-6 0.01-6 methoxy- cinnamate Butyl 0-3 0.01-3 0.01-3 0.01-3 Methoxy- dibenzoyl- methane Ensulizole 0-4 0.01-4 Octinoxate 0-7.5 Octisalate 0-5 0.01-5 0.01-5 Octocrylene 0-10 0.01-10 0.01-10 Homosalate 0-10 0.01-10 Dimethicone 0-10 0.01-10 0.01-10 Cyclomethicone 0-15 0.01-15 Fragrance 0-2 0-2 0-2 0-2 0-2 Glutamine/ 0.01-10 0.01-10 0.01-10 0.01-10 0.01-10 Sodium PCA Glycine 0.01-10 0.01-10 0.01-10 0.01-10 0.01-10 Cystine 0.001-2 0.001-2 0.001-2 0.001-2 0.001-2 Nicotinamide 0.05-10 0.05-10 Nicotinamide 0.05-10 riboside Isonicotinamide 0.05-10 0.05-10 N-methyl 0.05-10 Nicotinamide Nicotinyl myristate

TABLE-US-00017 TABLE II Water-in-oil topical lotions or creams WO-1 WO-2 WO-3 WO-4 Water To 100 To 100 To 100 To 100 Glycerine 0-70 1-70 1-70 Propylene glycol 0-5 0.01-5 Butylene glycol 0-5 0.01-5 0.01-5 Disteardimonium Hectorite 0.01-1 0.01-1 EDTA 0.01-.01 0.01-1 0.01-1 0.01-1 Preservative 0.02-2 0.02-2 0.02-2 0.02-2 TiO2 0-10 0.01-10 0.01-10 0.01-10 Colorant/pigment 0-5 0-5 0-5 0-5 TEA/Sodium Hydroxide/ 0-3 0.01-3 0.01-3 0.01-3 potassium Hydroxide Stearic acid 0-5 0.01-5 Isopropyl Myristate 0-10 Capric/Caprylic 0-10 0.01-10 Triglyceride C12-C15 alkyl benzoate 0-10 0.01-10 Mineral oil 0-10 Glyceryl stearate 0-5 Dimethicone copolyol 0-5 0.01-5 0.01-5 Cetyl PEG/PPG-10/1 0-5 0.01-5 Dimethicone Steareth-2 0-2 Sucrose Distearate 0-2 0.01-2 Cetyl alcohol 0-2 0.01-2 0.01-2 Ethyl hexyl 0-6 0.01-6 methoxycinnamate Butyl 0-3 0.01-3 0.01-3 0.01-3 Methoxydibenzoylmethane Ensulizole 0-4 0.01-4 Octinoxate 0-7.5 Octisalate 0-5 0.01-5 0.01-5 Octocrylene 0-10 0.01-10 0.01-10 Homosalate 0-10 0.01-10 Dimethicone 0-10 0.01-10 0.01-10 Cyclomethicone 0-40 0.01-40 0.01-10 Caprylyl methicone 0-10 0.01-10 0.01-10 Dimethicone crosspolymer 0-90 0.01-90 0.01-90 C30-C45 alkyl cetearyl 0.01-90 dimethicone crosspolymer Glycolic acid 0-10 0.01-10 KCl 0-5 0.01-5 0.01-5 0.01-5 Fragrance 0-2 0-2 0-2 0-2 Glutamine/Sodium PCA 0.01-10 0.01-10 0.01-10 0.01-10 Glycine 0.01-10 0.01-10 0.01-10 0.01-10 Cystine 0.001-2 0.001-2 0.001-2 0.001-2 Nicotinamide 0.05-10 Nicotinamide riboside 0.05-10 Isonicotinamide 0.05-10 N-methyl Nicotinamide 0.05-10

TABLE-US-00018 TABLE III Vanishing Creams VC-1 VC-2 VC-3 VC-4 Water To 100 To 100 To 100 To 100 Glycerine 0-5 0.01-5 0.01-5 EDTA 0.01-.01 0.01-.01 0.01-.01 0.01-.01 Preservative 0.02-2 0.02-2 0.02-2 0.02-2 TiO2 0.01-10 0.01-10 0.01-10 0.01-10 Colorant/pigment 0-5 0.01-5 0.01-5 TEA/Sodium Hydroxide/ 0-3 0.01-3 0.01-3 0.01-3 potassium Hydroxide Stearic acid 0-30 0.01-30 0.01-30 0.01-30 Isopropyl Myristate 0-5 0.01-10 0.01-10 C12-C15 alkyl benzoate 0-5 0.01-10 Brij 35 0-5 0.01-5 Tween40 0-5 0.01-5 Cetyl alcohol 0-2 0.01-2 0.01-2 Ethyl hexyl 0-6 0.01-6 0.01-6 methoxycinnamate Butyl 0-3 0.01-3 0.01-3 0.01-3 Methoxydibenzoylmethane Ensulizole 0-4 0.01-4 Octisalate 0-5 0.01-5 Octocrylene 0-10 0.01-10 0.01-10 Dimethicone 0-5 0.01-5 Cyclomethicone 0-5 0.01-5 Dimethicone crosspolymer 0-4 0.01-4 Niacinamide 0-5 0.01-5 0.01-5 0.01-5 Hydroxystearic acid 0-5 0.01-5 0.01-5 0.01-5 Fragrance 0-2 0-2 0-2 0-2 Glutamine/Sodium PCA 0.01-10 0.01-10 0.01-10 0.01-10 Glycine 0.01-10 0.01-10 0.01-10 0.01-10 Cystine 0.001-2 0.001-2 0.001-2 0.001-2 Nicotinamide 0.05-10 Nicotinamide riboside 0.05-10 Isonicotinamide 0.05-10 N-methyl Nicotinamide 0.05-10