WO2016064943A1 - Procédé de caractérisation de la capacité protectrice d'une substance de protection de la peau topique - Google Patents

Procédé de caractérisation de la capacité protectrice d'une substance de protection de la peau topique Download PDF

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Publication number
WO2016064943A1
WO2016064943A1 PCT/US2015/056555 US2015056555W WO2016064943A1 WO 2016064943 A1 WO2016064943 A1 WO 2016064943A1 US 2015056555 W US2015056555 W US 2015056555W WO 2016064943 A1 WO2016064943 A1 WO 2016064943A1
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score
composition
protective
protein
human skin
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PCT/US2015/056555
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English (en)
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Joseph Abernathy Lewis Ii
Arthur J. Pellegrino
Lavinia C. Popescu
Jose Ricardo FERNANDEZ
Eduardo Perez
Paul A. SOWYRDA
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Us Cosmeceutechs, Llc
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Publication of WO2016064943A1 publication Critical patent/WO2016064943A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • 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
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/66—Enzymes

Definitions

  • the present invention relates in general to topical protection capacity of substances and in particular to a method for characterizing the topical protective capacity of a substance.
  • Sun protection factor is a measure of a topical protective capacity of a substance such as a sunscreen against type B ultraviolet radiation (UV-B). The higher the SPF, the more protection a sunscreen offers against UV-B radiation, which is the radiation that causes sunburn. SPF labeling of sunscreen products is regulated by the U.S. Food and Drug Administration (FDA). SPF is an example of a standardized sun protection factor a substance. Determination of SPF values of products is described in Title 21 of the Code of Federal Regulations, specifically at 21 C.F.R. ⁇ 352.73, which is incorporated by reference herein.
  • UV-A radiation may cause DNA damage to cells which may increase the risk of malignant melanomas.
  • substances that provide protection against UV-A radiation include zinc oxide, oxybenzone, avobenzone, and meroxyl.
  • standards that have been developed for measuring the topical protective capacity of a substance against UV-A radiation include Critical Wavelength (FDA Broad Spectrum Claim), Persistent Pigment Darkening (PPD), Immediate Pigment Darkening (IPD), Boots Star System, and the Japanese PA system. These measurements are further examples of a sun protection values for a substance.
  • UPF ultraviolet protection factor
  • SPF ultraviolet protection factor
  • the UVA wavelengths (315 to 400 nm) of the solar ultraviolet spectrum are at least 10 times less effective at producing a sunburn, but readily penetrate the entire depth of the skin where a different kind of damage can occur, including structural damage to the dermal fibroblasts and dermal proteins such as collagen and elastin.
  • Higher SPF products usually incorporate both UVB and UVA sunscreen agents into the formulation, in which case the formulation is designated as a broad spectrum sunscreen; sunscreens in the US are now required to provide broad spectrum UV protection.
  • IR radiation represents over 50% of the radiation arriving at the earth's surface; currently US & INTL sun protection standards do not address the need for protection against IR radiation.
  • SPF skin protection The benefits of sunscreen have been studied for decades. Studies show that in the
  • SPF 15 blocks 94% of UVB radiation; SPF 30 blocks 97% of UVB radiation; and SPF 45 blocks 98% of UVB radiation.
  • Damage to human skin can be caused by oxidative stress from external environmental stressors, including sunlight radiation, but also from other non-radiation sources such as exposure to air pollution, particulate pollution, cigarette smoke, chemicals, cosmetics, drugs, ozone and even oxygen itself.
  • UV radiation is known to be the predominant cause of premature aging of the skin. UV radiation may lead to photochemical reactions, wherein then the photochemical reaction products intervene in the skin mechanism. Predominantly such photochemical reaction products are free-radical compounds, for example hydroxy radicals. Also, undefined free-radical photoproducts, which are produced in the skin itself, may trigger uncontrolled side reactions due to their high reactivity. Furthermore, lipid peroxidation products, such as for example hydroperoxides and aldehydes, are produced. Free-radical chain reactions may be triggered, resulting in skin damage.
  • UV radiation is known to be the predominant cause of premature aging of the skin. UV radiation may lead to photochemical reactions, wherein then the photochemical reaction products intervene in the skin mechanism. Predominantly such photochemical reaction products are free-radical compounds, for example hydroxy radicals. Also, undefined free-radical photoproducts, which are produced in the skin itself, may trigger uncontrolled side reactions due to their high reactivity. Furthermore, lipid peroxidation products,
  • UV radiation is ionizing radiation.
  • Sunlight provides an irradiance of just over 1 kilowatt per square meter at sea level, 527 watts of this energy being infrared radiation (IR), 445 watts being visible light, and 32 watts being ultraviolet radiation. Intense IR exposure to certain tissue can also be damaging.
  • oxidative stress may occur as a natural by-product of cellular energy production. Both internal and external oxidative stress leads to inflammatory pathways mediated by the formation of free radicals (molecules with unpaired electrons that are highly reactive) that, left unchecked, can cause severe cellular damage to cell membranes, lipids, proteins and DNA.
  • free radicals molecules with unpaired electrons that are highly reactive
  • the superoxide radical a natural by-product of metabolic energy production, causes serious deleterious effects to living cells if not quenched, neutralized or reduced almost immediately after production. It is known that lipid peroxidation is a major problem in biological systems. Protecting against cell membrane oxidation is of paramount importance in living biological systems since the cell membrane is the cell's first line of defense against oxidation
  • Antioxidants are substances, to include free radical absorbers or scavengers, that prevent oxidation processes, including oxidation of a molecule such as a lipid, lipoprotein, protein or DNA, or autooxidation of fats containing unsaturated compounds.
  • Various antioxidants are used in the field of cosmetics and pharmacy. These include, for example tocopherol, kinetin, ubiquinone, ascorbic acid, lipoic acid, sesamol, colic acid derivatives, butylhydroxy anisole and butylhydroxy toluene. Antioxidants can thus help protect human cells, such as skin cells, from both externally and internally generated oxidative stress.
  • CPD cyclobutane pyrimidine dimers
  • Known rating systems of measuring sun protection factors e.g., SPF, but also UPF, PPD, IPD, etc.
  • sun protection factors e.g., SPF, but also UPF, PPD, IPD, etc.
  • the known measurements for characterizing the oxidative stress protection capacity of a substance do not give any indication of the photo protective capacity of that substance.
  • a skin preparation may provide effective protection against photo damage to the skin but have little or no antioxidative qualities.
  • Other skin preparations may include effective antioxidants, but provide only marginal protection against UV photo damage to the skin. Relying on such measurement standards does not provide a consumer with the information he or she needs to evaluate the total topical protective capacity of a product.
  • SPF and antioxidant protection do not give any indication of the level of DNA protection because neither are direct measurements of DNA damage although both in theory provide DNA protection by their mechanism of protection/action.
  • TPF Total Protection Factor
  • GPP Total Genetic/Protein Protection
  • SPF Sun Protection Factor
  • EPF Environmental Protection Factor
  • GPP Genetic Protection Factor
  • TPF will reflect an enhanced score, or will capture the multiplicative protective effect, of synergistic combinations of EPF antioxidants, GPF repair enzymes, and the SPF sunscreen agents.
  • TPF can be reported to consumers in the same manner as SPF is reported, i.e., a numerical score, or as a set of scores corresponding to more than one, or each, type of protection tested.
  • An aspect of the invention provides a method for testing skin preparations for protection against photo (sun) damage, genetic damage, and oxidative damage to the skin.
  • An aspect of the invention involves measuring SPF (which may include one, two, three, four, five, or more tests) and GPF (which may include one, two, three, four, five, or more tests), and optionally further EPF (antioxidative efficacy, which may include one, two, three, four, five, or more tests) of a formulation.
  • the scores can be presented individually and/or combined, e.g., as raw, weighted, summed, averaged, and/or normalized scores or symbols or colors or other figures.
  • the SPF, EPF, and/or GPF scores can be aggregated in many ways, by linear addition, weighted addition, averaging, factoring, etc., to provide a single TPF value, which may be presented to consumers.
  • This type of TPF value may represent the efficacy of the total composition for sunburn prevention (SPF), antioxidant protection (EPF), and genetic protection (GPF).
  • the total TPF score may be based on a weighted combination of each of these scores.
  • the three scores may be set forth as a three part single presentation score, such as A, B, C... for sun protection (SPF), 1, 2, 3, 4, 5... for antioxidant protection (EPF), and 1, 2, 3, 4, 5... for genetic protection (GPF), to give, e.g., A45 or A-4-5 as the SPF-EPF-GPF value of the product.
  • An aspect of the invention provides a method of characterizing a topical genetic and/or protein protective capacity or sun, antioxidant, and/or genetic protective capacity, relative to human skin, of a composition, the method comprising: assigning a first score, indicative of a first protective capacity of a first component of the composition relative to human skin, to the first component of the composition; assigning a second score, indicative of a second protective capacity of a second component of the composition relative to human skin, to the second component of the composition; assigning a third score, indicative of a third protective capacity of a third component of the composition relative to human skin, to the third component of the composition, and presenting the scores, individually and/or optionally combining all scores, including the first score, second score, and third score, to obtain and optionally further present a total protective factor value, wherein the composition preferably comprises a non-antioxidative substance from which a tested component is selected, and optionally further testing an antioxidative substance, or optionally a sun screen, an antioxidant (or plurality thereof), and/
  • An aspect of the invention provides a method of characterizing a topical protective capacity, relative to human skin, of a substance, the method comprising: assigning to the substance at least a protein protection antioxidant score indicative of a protective capacity of the substance relative to human skin, the protein protection antioxidant score being based on a performance of the substance in reducing protein carbonylation, protein glycation, or both; assigning to the substance a genetic score indicative of a protective capacity of the substance relative to human skin, the genetic score being based on a performance of the substance in reducing cyclobutane pyrimidine dimer formation, 8-oxo-7,8-dihydro-2'-deoxyguanosine formation, or both; and presenting the scores, individually and/or optionally combining the protein protection antioxidant score and the genetic score, to obtain and optionally further present a total score indicative of a protective capacity of the substance relative to human skin, preferably additionally assessing the SPF protection of the substance.
  • Fig. 1 illustrates an embodiment wherein individual components of a composition are tested by a number, "N," of tests for, e.g., certain biological markers, whereby an aggregation, sum, or other compilation of results of each test can be compiled in the x-direction and/or an aggregation, sum, or other compilation of results of each component in one test can be compiled in the y-direction.
  • N a number of tests for, e.g., certain biological markers
  • An aspect of the invention provides a method of characterizing a protective capacity, relative to human skin, of a substance, the method comprising: (a) assigning to the substance a sun protection factor score indicative of a sunblocking efficacy of the substance relative to human skin; (b) assigning to the substance a antioxidative score, optionally a protein protection antioxidative score, indicative of an oxidative stress protective capacity of the substance relative to protein protection relative to human skin; (c) assigning to the substance a genetic score indicative of a genetic damage reduction or repair capacity of the substance relative to human skin, the genetic repair score evaluating at least cyclobutane pyrimidine dimer formation and 8-oxo-7,8-dihydro-2'-deoxyguanosine formation; and presenting the scores, individually and/or optionally combining the protein protection antioxidative score, the genetic repair score, and the sun protection factor score, to obtain and optionally further present a total score characterizing the protective capacity of the substance relative to human skin.
  • An aspect of the invention provides a method of characterizing a topical protective capacity, relative to human skin, of a substance, the method comprising: assigning to the substance at least a protein and genetic protection factor (SPF GPP) score indicative of a GPP protective capacity of the sunscreen substance relative to human skin.
  • SPF GPP protein and genetic protection factor
  • This analysis may include a protein protection sunscreen score being based on a performance of the sunscreen substance in reducing protein carbonylation, protein glycation, or both; assigning to the sunscreen substance a genetic sunscreen score indicative of a protective capacity of the substance relative to human skin, the genetic score being based on a performance of the substance in reducing cyclobutane pyrimidine dimer formation, 8-oxo-7,8-dihydro-2'- deoxyguanosine formation, or both; and presenting the scores, individually and/or optionally combining the protein protection antioxidant score and the genetic sunscreen score, to obtain and optionally further present a total SPF GPP score indicative of a protective capacity of the sunscreen substance relative to human skin.
  • An aspect of the invention provides a method of characterizing a topical protective capacity, relative to human skin, of a substance, the method comprising: assigning to the substance at least a protein and genetic (EPF GPP) score indicative of a GPP protective capacity of the antioxidant substance relative to human skin.
  • EPF GPP protein and genetic
  • This analysis may include a protein protection antioxidant score being based on a performance of the antioxidant substance in reducing protein carbonylation, protein glycation, or both; assigning to the antioxidant substance a genetic antioxidant score indicative of a protective capacity of the substance relative to human skin, the genetic score being based on a performance of the substance in reducing cyclobutane pyrimidine dimer formation, 8-oxo-7,8-dihydro-2'- deoxyguanosine formation, or both; and presenting the scores, individually and/or optionally combining the protein protection antioxidant score and the genetic antioxidant score, to obtain and optionally further present a total EPF GPP score indicative of a protective capacity of the antioxidant substance relative to human skin.
  • An aspect of the invention provides a method of characterizing a topical protective capacity, relative to human skin, of a substance, the method comprising: assigning to the substance at least a protein and genetic (DPF GPP) score indicative of a GPP protective capacity of the DNA/protein repair substance relative to human skin.
  • DPF GPP protein and genetic
  • This analysis may include a protein protection DNA/protein repair score being based on a performance of the DNA/protein repair substance in reducing protein carbonylation, protein glycation, or both; assigning to the DNA/protein repair substance a genetic DNA/protein repair score indicative of a protective capacity of the substance relative to human skin, the genetic score being based on a performance of the substance in reducing cyclobutane pyrimidine dimer formation, 8- oxo-7,8-dihydro-2'-deoxyguanosine formation, or both; and presenting the scores, individually and/or optionally combining the protein protection antioxidant score and the genetic DNA/protein repair score, to obtain and optionally further present a total DPF GPP score indicative of a protective capacity of the DNA/protein repair substance relative to human skin.
  • the SPF GPP, EPF GPP, and/or DPF GPP scores can be aggregated in many ways, by linear addition, weighted addition, averaging, factoring, etc., to provide a single TPF value, which may be presented to consumers.
  • This type of TPF value may represent the efficacy of each component for protecting genetic elements of the skin (proteins, nucleotides, etc.), even though the component may not primarily function as a genetic repair component, e.g., a sunscreen, which may not primarily function as a genetic repair component, versus a DNA repair enzyme, which may primarily function as a genetic repair component.
  • This type of TPF analysis may represent the combined genetic/protein protective contributions of each component of a formulation, and may be presented to, e.g., consumers, as a numerical (or other qualitative) value alongside other familiar values, such as SPF.
  • An aspect of the invention provides a method of characterizing protective capacity, relative to human skin, of a substance, the method comprising: (a) assigning to the substance a sun protection factor score indicative of a sunblocking efficacy of the substance relative to human skin; (b) assigning to the substance a protein protection antioxidative score indicative of an oxidative stress protective capacity of the substance relative to protein protection relative to human skin; (c) assigning to the substance a genetic score indicative of a genetic damage reduction or repair capacity of the substance relative to human skin, the genetic repair score evaluating at least cyclobutane pyrimidine dimer formation and 8-oxo-7,8- dihydro-2'-deoxyguanosine formation; and presenting the scores, individually and/or optionally combining the protein protection antioxidative score, the genetic repair score, and the sun protection factor score, to obtain and optionally further present a total score characterizing the protective capacity of the substance relative to human skin.
  • This type of TPF value may represent the efficacy of each component for protecting the skin, based upon the primary protective function of the respective component, e.g., a sunscreen, evaluated for its blockage/dissipation of UV irradiation; a DNA repair enzyme, evaluated for its efficacy in repairing DNA; an antioxidant, evaluated for its absorption/quench/extinction of oxidizers.
  • This type of TPF analysis may represent the combined contributions of each component to the total protection of a formulation, and may be presented to, e.g., consumers, as a numerical (or other qualitative) value, preferably in place of other values, such as SPF, EPF, etc.
  • An aspect of the invention provides a method of characterizing cumulative protective capacity, relative to human skin, of a substance, preferably a composition comprising a sunscreen agent, antioxidant substance(s), and DNA/'protein repair enzyme(s), the method comprising the additive or multiplicative GPP scoring of: (a) assigning to the sunscreen agent a SPF GPP score indicative of the efficacy of the substance relative to human skin in protecting against genetic/protein damage; (b) assigning to the antioxidant substance(s) a EPF GPP score indicative of the efficacy of the substance relative to human skin in protecting against genetic/protein damage; (c) assigning to the DNA/protein repair substance a DPF GPP score indicative of the efficacy of the substance relative to human skin in protecting against genetic/protein damage, the genetic repair score evaluating at least cyclobutane pyrimidine dimer formation and 8-oxo-7,8-dihydro-2'-deoxyguanosine formation; and presenting the scores, individually and/or optionally measuring/combining SPF GPP,
  • An aspect of the invention provides a method of characterizing a topical protective capacity of a substance suitable for application to human skin, alone or combined with any of the steps above, the method comprising: assigning to the substance a sun protective score indicative of a sun protection factor of the substance for protecting human skin against ultraviolet light; assigning to the substance a protein protection antioxidant score indicative of a protein damage reduction capacity of the substance relative to human skin; assigning to the substance a genetic score indicative of a genetic damage reduction or repair capacity of the substance relative to human skin, the genetic score evaluating at least one of cyclobutane pyrimidine dimer formation and 8-oxo-7,8-dihydro-2'-deoxyguanosine formation; and presenting the scores, individually and/or optionally combining the first, second, and third scores, to obtain and optionally further present a total score indicative of a protective capacity of the substance relative to human skin.
  • An aspect of the invention provides a method for informing a purchaser of a substance of a topical protective capacity of the substance, the method comprising: providing the total score indicative of the topical protective capacity of the substance, obtained by any of the above methods; and making an information including the score available to the purchaser.
  • the "making available" of any of the above scores or the TPF value may be achieved by printing and attaching a label to an object; entering the value into a database which may operate in a digital system to project the data onto a display, personal or public; printing a poster or analog display; etching into a surface; etc.
  • the evaluation may involve any number of analyses, particularly reduction of cyclobutane pyrimidine dimer (CPD) formation, protein carbonylation (PC), 8- oxo-7,8-dihydro-2'-deoxyguanosine formation (80HdG) formation, DNA/RNA alkylation, protein alkylation (including glycation), etc.
  • the tests may be carried out (a) on singular substances, i.e., an amount of a single molecule, or (b) on a plurality (2, 3, 4, 5, etc.) of molecules of a single class (for example, antioxidants, protein protection antioxidants, repair enzymes, protein repair enzymes, DNA repair enzymes, RNA repair enzymes, etc.), or (c) on a composition including all of its components.
  • each component/class may be aggregated as desired and, optionally, all of the components/classes may be aggregated.
  • Appropriate weight may be used, as may be additive or multiplicative approaches.
  • An aspect of the invention provides a method for marketing a topical skin preparation, the method comprising providing the skin care preparation containing a substance; and providing information including a topical protective score indicative of a topical protective capacity of the skin care product, obtained by any of the above methods.
  • An aspect of the invention provides a method of analyzing a substance for preventing, reducing, or remedying genetic and protein damage to the skin caused by environmental stress.
  • the analysis of genetic and protein damage preferably takes into consideration one or more of cyclobutane pyrimidine dimer (CPD) formation, 4-6 photoproduct formation, 8-oxo- 7,8-dihydro-2'-deoxyguanosine (80HdG) formation, nucleotide alkylation, protein carbonylation (PC), protein glycation, and protein alkylation.
  • CPD cyclobutane pyrimidine dimer
  • 80HdG 8-oxo- 7,8-dihydro-2'-deoxyguanosine
  • the genetic damage analysis includes each of a cyclobutane pyrimidine dimer (CPD) formation and an 8-oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) formation evaluation
  • the protein damage analysis includes a protein carbonylation (PC) and/or anti-glycation evaluation.
  • the analytical results may be combined in a manner that represents the best efficacy for the targeted markers (e.g., CPD, 80HdG, and/or PC).
  • An aspect of the invention allows that any one, two, three, four, five, or six of a cyclobutane pyrimidine dimer (CPD) analysis, 8-oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) analysis, and/or nucleotide alkylation analysis be combined with one, two, three, four, five, six, seven, or more antioxidation analyses including, e.g., protein carbonylation (PC) analysis, protein glycation analysis, and/or protein alkylation analysis.
  • CPD cyclobutane pyrimidine dimer
  • 80HdG 8-oxo-7,8-dihydro-2'-deoxyguanosine
  • nucleotide alkylation analysis be combined with one, two, three, four, five, six, seven, or more antioxidation analyses including, e.g., protein carbonylation (PC) analysis, protein glycation analysis, and/or protein alkylation analysis.
  • a method involving, e.g., CPD and/or 80HdG analysis, optionally with further genetic analysis, combined with singular or composite protein protection antioxidant analysis (protein carbonylation (PC) and/or glycation) is preferred in certain embodiments of the invention.
  • PC protein carbonylation
  • glycation singular or composite protein protection antioxidant analysis
  • an compound or composition under study can be subjected to any 1, 2, 3, 4, 5, or more independent antioxidative efficacy tests, in addition to 1, 2, 3, 4, 5, or more independent genetic repair analyses.
  • a method involving genetic analysis (1, 2, 3, 4, 5, 6, or more tests), combined with protein protection antioxidant analysis (1, 2, 3, 4, 5, 6, 7, 8, or more tests), may be combined with a traditional sun protection factor (SPF) analysis and/or antioxidant capacity analyses in certain embodiments, to give a broader appraisal of the protective strength of the substance.
  • SPF sun protection factor
  • the genetic damage/repair analysis preferably includes testing for DNA and/or NA damage.
  • the genetic analysis preferably includes testing for each of cyclobutane pyrimidine dimer (CPD) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) formation.
  • the protein protection analysis preferably includes protein carbonylation (PC) and/or protein glycation analysis. Certain embodiments may substitute or supplement one or more of the
  • RNA molecular damage RNA molecular damage
  • DNA molecular damage DNA molecular damage
  • protein carbonylation protein glycation
  • Cyclobutane pyrimidine dimer (CPD) formation is UV-induced DNA helix distortion believed to play a role in photocarcinogenesis.
  • CPDs are increased in the skin of patients with non-melanoma skin cancer (NMSC) and can cause immunosuppression, potentially allowing cancer growth while decreasing immunosurveillance.
  • 8-Oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) is a main product of the oxidation of double-stranded DNA because guanine, with the lowest oxidation potential, and is most easily oxidized.
  • 8-oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) formation increases in UV-irradiated human skin in a dose-dependent manner, thus serving as a potential indicator antioxidant photoprotective efficacy in human skin.
  • PC protein carbonylation
  • Protein carbonylation studied within the scope of the inventive analysis is a type of protein oxidation that can be promoted by reactive oxygen species, typically in a process that forms reactive ketones or aldehydes that can be react, i.e., with protein side chains and/or other reactive moieties.
  • reactive oxygen species typically in a process that forms reactive ketones or aldehydes that can be react, i.e., with protein side chains and/or other reactive moieties.
  • Examples of this are the direct oxidation, via carbonylation, of side chains of lysine, arginine, proline, and threonine residues, among other amino acids.
  • Protein glycation studied within the scope of the inventive analysis is preferably non- enzymatic glycation, e.g., UV- or heat-induced glycation.
  • Protein glycation involves the reaction of a sugar, e.g., as glucose or fructose, with the amino group of a protein to form a Schiff base. This process can occur even under physiological conditions, and has been associated with disease, such as diabetes, and with aging.
  • One approach to analyzing antioxidant capacity of a substance for the skin can be to compare the formation of glycation modifications to one or more proteins in an untreated, or alternately treated subject, in with the reduction of modifications in the case of using no protective agent (i.e., plain skin) or using a protective agent or mixture of agents.
  • alkylation is another manner by which nucleotides and proteins can become damage and suffer diminished or complete loss of function. This can occur by either chemical or biological agents and may be used as a marker for the genetic protection of skin from environmental stress.
  • DNA, RNA, and/or protein damage may be analyzed by any number of methods known or readily apparent to those of ordinary skill in the art.
  • analytical techniques examples include two-dimensional gel electrophoresis, followed by immunoblotting and/or mass spectrometry, chromatography (e.g., HPLC, LC, MPLC) followed by mass spectrometry, mass spectrometry techniques alone or combined with separative techniques, and/or more or more spectroscopies, such as UV-vis, IR, fluorescence, ESR, and/or NMR.
  • one, two, three, four, five, or more of the antioxidative tests below are preferably included in a composite or multi-composite analysis.
  • This genetic and antioxidative analysis may be further combined with a sun protection factor (SPF) test score.
  • SPF sun protection factor
  • any combination of evaluative scores from the one, two, three, or more genetic repair analysis may be combined with a score from a sun protection factor (SPF) test to give an alternate analytical result.
  • nucleotide damage/repair e.g., DNA and/or RNA damage/repair
  • Certain preferred embodiments of the genetic damage evaluation take into consideration, for example, 8-oxo- 7,8-dihydro-2'-deoxyguanosine (80HdG) formation, and/or cyclobutane pyrimidine dimer (CPD) formation, preferably both, and optionally more.
  • 80HdG 8-oxo- 7,8-dihydro-2'-deoxyguanosine
  • CPD cyclobutane pyrimidine dimer
  • Each genetic damage assessment can be used to give a single, double, triple, quadruple, etc., genetic protection score as a composite genetic protection score, which is preferably combined with a protein protection antioxidative evaluative score (which can also be a double, triple, quadruple, quintuple, etc., antioxidative score as a composite antioxidative score), and more preferably further includes a sun protection factor (SPF) score.
  • a protein protection antioxidative evaluative score which can also be a double, triple, quadruple, quintuple, etc., antioxidative score as a composite antioxidative score
  • SPF sun protection factor
  • Any of these scores can include multiple "dimensions," i.e., a 1, 2, 3, 4, 5, ... 10-factor genetic protection score, and a 1, 2, 3, 4, 5, ...
  • 10-factor antioxidative score which may be combined with each other in any iteration, preferably combined with an sun protection factor (SPF) score in a meaningful way such that the qualitatively best material is apparent from the sum.
  • SPF sun protection factor
  • CPD cyclobutane pyrimidine dimer
  • 80HdG 8-oxo-7,8-dihydro-2'-deoxyguanosine
  • PC protein carbonylation
  • the following antioxidants were tested based on three genetic markers tests: quercetin, L- carnosine, resveratrol, L-ergothioneine, ferulic acid, a-tocopherol, chlorogenic acid, N-acetyl- S-farnesyl-L-cysteine, phloretin, ascorbic acid, epigallocatechin gallate (EGCG), 2-allyl- pyrroloquinoline quinone (2-allyl-PQQ),
  • the makers can include protein glycation, lipid glycation, protein alkylation, DNA alkylation, RNA alkylation, and/or lipid alkylation.
  • the genetic markers can be analyzed individually, or preferably, in combination, to assess the genetic protective capacity of each antioxidant.
  • the results of each of the three tests for each antioxidant can be combined so as to provide a total score indicative of overall performance of each antioxidant.
  • Any antioxidant, and even non-antioxidants can be scored in each genetic and protein protection test, and the results then totaled for each substance for each of the tests on an equal weighted basis.
  • the individual tests can produce a score, and that score can be combined with other scores using a properly weighted basis.
  • the performance of an antioxidant with the best result relative to the results of the other antioxidants in a given test may serve as the standard for that test.
  • the total score of a particular antioxidant is preferably reflective of the overall genetic stress protective capacity of the antioxidant.
  • the genetic score can also be based on an arbitrary standard.
  • further embodiments involve adding one, two, three, four, five, or more further genetic analysis scores to the three discussed above. Another approach would be to average the scores, or weight the scores appropriately, such that the efficacy of any substance falls within a preferred range, e.g., 0 to 100.
  • the spectral irradiance was measured with an OL754TM spectroradiometer (Optronics, Orlando, FL, USA), calibrated for wavelength and intensity against standard lamps.
  • the spectroradiometer was used to calibrate a handheld IL700TM radiometer (International Light, Newburyport, MA), which was then used to rapidly monitor lamp output on a daily basis.
  • the minimal erythema dose was determined for each individual for solar- simulated UVR (290-400 nm) and expressed in mj/cm 2 using a light-proof adhesive-backed foil template that were sequentially uncovered to deliver quantities of UV above and below the expected MED of skin phototype I-II individuals for solar-simulating UV radiation.
  • the sites were examined 24 hours after irradiation and the MED was determined as the site that showed minimal, uniform perceptible erythema.
  • PC Protein carbonylation
  • Proteinase K was then heat inactivated at 95°C for 10 minutes, and homogenates were extracted using the PUREGENETM DNA isolation kit (Gentra Systems, Minneapolis, MN, USA) containing two main reagents: cell lysis and protein precipitation solutions.
  • DNA was extracted from homogenates using a lysis buffer solution and then treated with RNase A.
  • the kit removes proteins using a precipitation solution, followed by 2- propanol to pellet the DNA.
  • Cyclobutane pyrimidine dimers (CPD) and 8-oxo-7,8-dihydro- 2'-deoxyguanosine (80HdG) can be measured in duplicate and random order by specific ELISA kits [OXISELECTTM Cellular UV-Induced DNA Damage ELISA Kit (CPD), 96 assays (Cell Biolabs, San Diego, CA, USA)] and OXISELECTTM Oxidative DNA Damage ELISA Kit (8-OHdG Quantitation, Cell Biolabs, San Diego, CA, USA) according to the manufacturer's protocol.
  • cyclobutane pyrimidine dimers CPD
  • 8-oxo-7,8- dihydro-2'-deoxyguanosine 80HdG
  • PC protein carbonylation
  • An aspect of this invention provides combined score of protection against protein carbonylation, repair of DNA damaged by cyclobutane pyrimidine dimer (CPD) formation, and repair of DNA damaged by 8-oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) formation as shown below in Table 1.
  • CPD cyclobutane pyrimidine dimer
  • 80HdG 8-oxo-7,8-dihydro-2'-deoxyguanosine
  • the genetic analysis preferably takes into consideration the counteraction and/or repair of cyclobutane pyrimidine dimer (CPD) formation, 8-oxo-7,8-dihydro-2'- deoxyguanosine (80HdG) formation, and/or nucleotide alkylation.
  • CPD cyclobutane pyrimidine dimer
  • 80HdG deoxyguanosine
  • a system of evaluation of protection may be that described in our own work, US 2006/0171886 Al, which is incorporated by reference herein, optionally modified to include a genetic, protein, DNA, or RNA damage analysis, based preferably on the specific markers cyclobutane pyrimidine dimer (CPD) and/or 8-oxo-7,8-dihydro-2' -deoxyguanosine (80HdG) formation.
  • Results of an evaluation of DNA repair for certain substances are shown below in Table 2.
  • Table 2 Normal Human Epidermal Keratinocytes (NHEK), UV
  • the analysis of protein protective capacity preferably takes into consideration the protection from protein carbonylation (PC), glycation, lipidation, and/or alkylation, and may alternately or additionally measure lipid glycation and/or alkylation.
  • PC protein carbonylation
  • US 2006/0171886 Al offers an effective antioxidant analytical method which may be modified to evaluate protein carbonylation, protein glycation, and/or protein alkylation, to appraise the overall protective quality of substances. Test results for selected antioxidants in an embodiment of the invention for the protection of human skin against protein carbonylation and UVB damage are shown below in Table 3.
  • antioxidants were tested according to all five tests: idebenone (6-(10-hydroxydecyl)-2,3-dimethoxy-5-methyl-l,4-benzoquinone); dl-alpha tocopherol (vitamin E); kinetin (a plant derivative); ubiquinone (coenzyme Q-10); 1-ascorbic acid (vitamin C); and dl-alpha lipoic acid.
  • the five tests can be used individually, or preferably, in combination, to assess the oxidative stress protective capacity of each antioxidant.
  • the results of each of the five tests for each antioxidant can be combined so as to provide a total score indicative of overall performance of each antioxidant.
  • the five tests are set forth in U.S.
  • One or more of the above antioxidant tests may be performed in any permutation, e.g.: Tests 1 and 2; Tests 4 and 16; Tests 8 and 9; any two tests; Tests 3, 4, and 7; any three tests, any four tests, any five tests; ... all tests except one; or all tests; and, for example, combined and/or set comparatively together with the results of the DNA, RNA, and/or further protein (genetic) damage analyses, e.g., any permutation of genetic tests listed below, and/or combined with an SPF score.
  • any permutation e.g.: Tests 1 and 2; Tests 4 and 16; Tests 8 and 9; any two tests; Tests 3, 4, and 7; any three tests, any four tests, any five tests; ... all tests except one; or all tests; and, for example, combined and/or set comparatively together with the results of the DNA, RNA, and/or further protein (genetic) damage analyses, e.g., any permutation of genetic tests listed below, and/or
  • antioxidant tests 1-16 may be combined with any one, two, three, four, five, six, or more of a cyclobutane pyrimidine dimer (CPD) analysis, 8-oxo-7,8-dihydro-2'-deoxyguanosine (80HdG) analysis, nucleotide alkylation analysis, protein carbonylation (PC) analysis, protein glycation analysis, protein alkylation analysis, and/or further DNA, RNA, and/or protein biomarker analyses.
  • CPD cyclobutane pyrimidine dimer
  • 80HdG 8-oxo-7,8-dihydro-2'-deoxyguanosine
  • PC protein carbonylation
  • protein glycation analysis protein alkylation analysis
  • protein alkylation analysis and/or further DNA, RNA, and/or protein biomarker analyses.
  • the results may be combined and/or presented in a manner that represents the best efficacy for the targeted biomarkers.
  • This test assessed DNA damage in cell culture experiments under pro-oxidative conditions (UVB irradiation of human keratinocytes) by measuring the positive cells for anti-thymine dimer antibodies. This experiment is thought to reflect a direct correlation to the in vivo occurring DNA cross linking damage following UVB exposure and the protection of such nuclear damage by antioxidants.
  • UVB Ultraviolet Light
  • Keratinocyte cultures were irradiated with a single dose of 200 mJ/cm 2 UVB, using FS-20/T-12 bulbs (emission range: 305-312 nm). Immediately prior to irradiation, the medium was replaced with 0.25 ml sterile PBS (pH 7.4, 37°C), and after irradiation, PBS was replaced with fresh growth medium. UVB exposure was quantified using a DAAVLINTM Flex Integrating Dosimeter Controller. Cells were maintained at 37°C (5% CO 2 ) for 1 hour until fixation with paraformaldehyde (PFA).
  • PFA paraformaldehyde
  • This test assessed DNA damage in cell culture experiments under pro-oxidative conditions (UVB irradiation of human keratinocytes) by measuring the positive cells for anti-8-oxoguanine antibodies. This experiment is thought to reflect a direct correlation to the in vivo occurring DNA cross linking damage following UVB exposure and the protection of such nuclear damage by antioxidants.
  • UVB Ultraviolet Light
  • Keratinocyte cultures were irradiated with a single dose of 200 mJ/cm 2 UVB, using FS-20/T- 12 bulbs (emission range: 305-312 nm). Immediately prior to irradiation, the medium was replaced with 0.25 ml sterile PBS (pH 7.4, 37°C), and after irradiation, PBS was replaced with fresh growth medium. UVB exposure was quantified using a DAAVLINTM Flex Integrating Dosimeter Controller. Cells were maintained at 37°C (5% CO2) for 1 hour until fixation with paraformaldehyde (PFA).
  • PFA paraformaldehyde
  • This test assessed protein damage in cell culture experiments under pro-oxidative conditions (UVB irradiation of human keratinocytes) by measuring the levels of total protein carbonylation (PC). This experiment is thought to reflect a direct correlation to the in vivo occurring protein carbonylation damage following UVB exposure and the protection of such protein damage by antioxidants.
  • UVB Ultraviolet Light
  • Keratinocyte cultures were irradiated with a single dose of 200 mJ/cm 2 UVB, using FS-20/T- 12 bulbs (emission range: 305-312 nm). Immediately prior to irradiation, the medium was replaced with 1 ml sterile PBS (pH 7.4, 37°C), and after irradiation, PBS was replaced with fresh growth medium. UVB exposure was quantified using a DAAVLIN Flex Integrating Dosimeter Controller. Cells were maintained at 37°C (5% C02) for 1 hour until total protein extraction.
  • cellular protein extracts were obtained by mechanical disruption and stored at -80°C.
  • the protein carbonylation colorimetric assay kit was obtained from Cayman Chemical Co. (Ann Arbor, MI). Protein carbonylation was quantified by production of hydrazone after protein carbonyl groups reacted with 2,4- dinitrophenylhydrazine (DNPH).
  • DNPH 2,4- dinitrophenylhydrazine
  • the testing involved the suppression of optical density (OD) at 340 ⁇ 25 nm to a degree that is proportional to an effective oxidation inhibition.
  • One or more genetic and/or protein protective tests may be performed, i.e.:
  • Test 1 and Test 2 Test 2 and Test 3; Test 1 and Test 3; Test 1, Test 2, and Test 3;
  • an antioxidative efficacy test may be any test that directly or indirectly provides information regarding the oxidative stress protective capacity of an antioxidative substance tested.
  • the oxidative stress protective capacity of an antioxidative substance may in some cases be function, for example, of its ability to prevent an environmental stressor from reaching and/or penetrating the skin. In other cases, the oxidative stress protective capacity of an antioxidative substance may, for example, be a function of its ability to prevent or reduce an oxidation process in the skin.
  • Such anti-oxidants include, for example, one or more (e.g., 2, 3, 4, 5, etc.) ofN-acetyl- 1 S'-[(2 J E,6 )-3,7, l l-trimethyl-2,6,10-dodecatrien-l-yl]- L-cysteine (N-acetyl-S-farnesyl-L-cysteine), L-ergothioneine, idebenone, L-carnosine, 2- allyl-pyrroloquinoline quinone (2-allyl PQQ), pyrroloquinoline quinone (PQQ), quercetin, resveratrol, epigallocatechin gallate, L-ascorbic acid, and ferulic acid.
  • N-acetyl- 1 S'-[(2 J E,6 )-3,7, l l-trimethyl-2,6,10-dodecatrien-l-yl]- L-cysteine N-acety
  • a protein protection antioxidant comprising N-acetyl-S-farnesyl-L-cysteine, L- carnosine, L-ergothioneine, or a mixture thereof, or comprising N-acetyl-5-farnesyl-L- cysteine, L-carnosine, and L-ergothioneine.
  • suitable protein protective antioxidants for use together with the above, or separately, in combination or alone, are set forth in U.S. Appl. Serial No. 62/066,425.
  • One embodiment of the invention evaluates the performance of an antioxidant or composition in a singular or composite antioxidant score, and the antioxidant score is combined, using a properly weighted basis, e.g., with one or more further antioxidant scores, preferably a protein protection antioxidant score, with an singular or composite genetic score (optionally weighted individually or compositely), and/or the optionally weighted SPF score. These scores can be reported as a set of values, two or more (different) sets of values, or a single value. In another embodiment, the performance of an antioxidant with the best result relative to the results of the other antioxidants in a given antioxidative efficacy test served as the standard for that test.
  • idebenone produced the greatest percent reduction in sun burn cells (SBCs) in the sunburn cell assay (Test 1) and its performance was selected as the standard, receiving 20 points. Kinetin performed better than the other antioxidants in the primary oxidative products test (Test 3), so kinetin' s performance was selected as the standard, receiving 20 points in that test. In other embodiments of the present invention, the performance of the same antioxidative substance, such as idebenone, irrespective of whether or not its performance is the best, may be selected as the standard in every test performed.
  • the performance of a single substance subjected to one or more tests relative to a standard, or to a respective standard for each test may be used to assign a score indicative of the protective capacity of the substance.
  • the scores from the tests may be summed to give a total score. Results or scores may be presented together or combined in any of a variety of ways to give a total score characterizing a substance is within the scope of the invention.
  • the tests may be on a composition, components of the composition, or both.
  • the ability of the composition, a component, or class of components— just as for AO or genetic tests, to protect skin from one or more, or all, types of (esp. sun blocking, for sun-generated) radiation may be tested.
  • This may include a capacity for protecting human skin from ultra-violet, visible, and/or infra-red light, but also microwaves, radio waves, telecommunication waves, atomic and subatomic particles, x rays, gamma rays, and/or any other form of radiation damaging, or potentially damaging, to the skin or tissue.
  • An aspect may apply MED and broad-band tests alone, or in combination with one or more tests for any preceding type of radiation.
  • Manners of combining the scores may include giving the sum of the scores in the tests, giving each score separately, an average of the scores, different weighted averages, though the weighting may be tailored in any way, e.g., polynomials, square roots of sums of squares, and other complex operations, to best capture the total protective capacity of a substance or composition, combined as a compound expression of the two scores separated by a dash, etc.
  • Other compound expressions for example, with the OSPC, CPD, 80HdG, or PC being the first score in the expression are also contemplated.
  • Other weights may be given to any of the scores in order give more or less weight of one of the components to the total score over the other component.
  • a variety of scoring and weighting methods may be employed in other embodiments of the present invention.
  • alphabetical, alpha-numeric or other symbological or graphical indicators such as pie chart, bar graphs, or numbers of stars, numbers of "smiley faces," etc.
  • the weighting of the test results could be adjusted, for example, for optimal clinical correlation based on the specific tests or experiments performed.
  • Table 4 Exemplary Assessment Combining a Partially Prophetic SPF Score, a DNA Repair Score, and a Protein Protection Antioxidant Score for a Composition Comprising Sunscreens, Nucleotide/Protein Repair Enzymes, and Antioxidants
  • Table 5 Exemplary Assessment Combining DNA Repair Scores of a Formulation Containing Non-Antioxidant Substances and Antioxidant Substances Based on Normalized Absolute Score
  • test results shown above in Table 5 an evaluation of assay results of a composition having both antioxidant substances and non-antioxidant substances, using normalized absolute scores, testing for a selection of DNA and/or protein markers. Note that each component makes a contribution to inhibiting deleterious markers, and the impact of this inhibition can be surprising.
  • Table 6 Exemplary Assessment Combining DNA Repair Scores of a Formulation Containing Non-Antioxidant Substances and Antioxidant Substances Based on Percent Improvement
  • test results shown above in Table 6 an alternate evaluation of assay results of a composition having both antioxidant substances and non-antioxidant substances, e.g., an antioxidant (preferably a plurality of protein protection antioxidants), a sunscreen agent (one or more sunscreens), and DNA/protein repair enzymes (preferably a plurality of repair enzymes), using relative percent improvement scores versus a carrier, testing for a selection of DNA and/or protein markers.
  • an antioxidant preferably a plurality of protein protection antioxidants
  • sunscreen agent one or more sunscreens
  • DNA/protein repair enzymes preferably a plurality of repair enzymes
  • Table 7 illustrates manners of combining scores to provide a total SPF score for the composition and/or component, a total EPF score for the composition and/or component, a total GPF score for the composition and/or component, which may be combined into one and/or presented categorically as several TPF scores for the composition and/or component.
  • Scores of individual components may be additive in certain circumstances, although the component-wise additivity of several different agents (or agents within one compound class, e.g., repair enzymes) ordinarily would not be expected in the experience of the inventors.
  • the selection of the appropriate markers to be tested can be important to delivering an accurate characterization of the protective capacity of the substance or composition. Weighting, averaging, appropriate normalizing (to a meaningful standard), or the like can improve the accuracy of summed scores for individual components.
  • a useful embodiment simply tests a total composition, for example, containing a mixture of antioxidants, DNA/protein repair enzymes, and sunscreens, for performance regarding one or more markers, for example, reduction of 80HdG, CPD, PC, DNA alkylation, protein alkylation (or glycation), and/or RNA alkylation.
  • the score(s) may be affixed to a label, displayed electronically (e.g., on an LCD or LED screen, or on a wall or screen with a projector), or included in a print presentation, such as printed onto a magazine page or a billboard, or a sheet to be applied to a surface.
  • the score(s) may be loaded into a database, entered into a computer device for further calculation or print-out, written and/or fixed onto a digital medium, or transmitted to a cloud.
  • the score(s) may be written onto paper or plastic (e.g., a label or packaging), etched (e.g., chemically or with laser) into plastic (e.g., a product container, a label, a display board), combined with other scores, or affixed as a set of 2, 3, 4, 5, or more, scores, in comparative tables, charts, etc.
  • plastic e.g., a label or packaging
  • etched e.g., chemically or with laser
  • plastic e.g., a product container, a label, a display board
  • individual test scores and a total score can be assigned to each or all antioxidants, sunscreen agents, and DNA/protein repair enzymes.
  • the protective capacity of topical preparation comprising antioxidant(s), sunscreen agent(s), and/or DNA/protein repair enzyme(s), as a whole may be characterized by performing one or more DNA/protein repair (i.e., for 80HdG, CPD, PC, alkylation, glycation, etc), optionally also with antioxidative and/or sunscreen efficacy tests on the entire preparation, rather than on individual (or component class-wise) antioxidative(s), genetic repair enzyme(s), and/or sunscreen substance(s).
  • an overall score or set of scores can be assigned to a topical skin preparation, indicative of the DNA/protein repair, or a combination of antioxidative, genetic repair, and/or sunscreen capacity of the preparation as a whole.
  • Such overall scores assigned to topical skin preparations enable an easy, effective way for consumers to compare the protective capacity of the preparations.
  • topical skin preparations may contain additional components that may be tested as compositions and/or as single-components.
  • topical skin preparations may contain cosmetic auxiliaries, as are used conventionally in such preparations, such as preservatives, bactericides, perfumes, substances for preventing foaming, dyestuffs, pigments which have a coloring effect, thickening agents, surfactant substances, emulsifiers, softening, moisturizing and/or moisture-retaining substances, fats, oils, waxes or other conventional constituents of a cosmetic or dermatological formulation, such as alcohols, polyols, polymers, foam stabilizers, electrolytes, organic solvents or silicone derivatives.
  • cosmetic auxiliaries as are used conventionally in such preparations, such as preservatives, bactericides, perfumes, substances for preventing foaming, dyestuffs, pigments which have a coloring effect, thickening agents, surfactant substances, emulsifiers, softening, moisturizing and/or moisture-retaining
  • protective compositions may include one or more DNA repair enzyme, preferably photolyase, endonuclease, 8-oxoguanine glycosylase, alkylase ⁇ e.g., alkylguanine- DNA alkyl transferase), or a mixture thereof, as these are suited for analysis as particularly preferred markers indicating skin damage.
  • Enzymes analyzed may be plant, animal, fungal, protozoan ⁇ rotist, yeast, archaeal, and/or bacterial in derivation ⁇ e.g., human, yeast, algae, plankton, etc.), and may be a synthetic variant thereof or a combination of natural and synthetic species. Further suitable repair enzymes are set forth in U.S. Appl. Serial No. 62/066,425.
  • the recitation of "at least one of A, B, and C” should be interpreted as one or more of a group of elements consisting of A, B, and C, and should not be interpreted as requiring at least one of each of the listed elements A, B, and C, regardless of whether A, B, and C are related as categories or otherwise.
  • the recitation of "A, B, and/or C" or "at least one of A, B, or C” should be interpreted as including any singular entity from the listed elements, e.g., A, any subset from the listed elements, e.g., A and B, or the entire list of elements A, B, and C.

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Abstract

La présente invention concerne des procédés de caractérisation d'une capacité protectrice topique, vis-à-vis d'une peau humaine, d'une composition, le procédé mettant en œuvre : l'attribution d'un premier score, indicatif d'une première capacité protectrice d'une composition vis-à-vis de la peau humaine, à la composition ; l'attribution d'un deuxième score, indicatif d'une deuxième capacité protectrice de la composition vis-à-vis de la peau humaine, à la composition ; l'attribution d'un troisième score, indicatif d'une troisième capacité protectrice de la composition vis-à-vis de la peau humaine, à la composition, et la présentation des scores et/ou la combinaison de l'ensemble des scores pour obtenir et/ou présenter une valeur de facteur de protection total, où la composition comporte une substance non antioxydante parmi laquelle un composant testé est sélectionné, les capacités protectrices comprenant facultativement la formation de dimère de cyclobutane-pyrimidine, la formation de 8-oxo-7,8-dihydro-2'-désoxyguanosine, la carbonylation de protéine, SPF et/ou une capacité antioxydante.
PCT/US2015/056555 2014-10-21 2015-10-21 Procédé de caractérisation de la capacité protectrice d'une substance de protection de la peau topique WO2016064943A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112881333A (zh) * 2021-01-13 2021-06-01 江南大学 一种基于改进免疫遗传算法的近红外光谱波长筛选方法
CN115919673A (zh) * 2022-12-19 2023-04-07 华熙生物科技股份有限公司 一种具有抗氧化作用的组合物及其用途

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060171886A1 (en) * 2005-02-03 2006-08-03 Pcr Technology Holdings, Lc Method for characterizing the oxidative stress protective capacity of an antioxidative substance
US20080056995A1 (en) * 2006-07-11 2008-03-06 Timmins Graham S System and methods for measuring a skin protection factor
WO2014037561A2 (fr) * 2012-09-10 2014-03-13 Laboratorios Cinfa, S.A. Composition antivieillissement de la peau
US20140234242A1 (en) * 2009-08-28 2014-08-21 Mary Kay Inc. Skin care formulations

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060171886A1 (en) * 2005-02-03 2006-08-03 Pcr Technology Holdings, Lc Method for characterizing the oxidative stress protective capacity of an antioxidative substance
US20080056995A1 (en) * 2006-07-11 2008-03-06 Timmins Graham S System and methods for measuring a skin protection factor
US20140234242A1 (en) * 2009-08-28 2014-08-21 Mary Kay Inc. Skin care formulations
WO2014037561A2 (fr) * 2012-09-10 2014-03-13 Laboratorios Cinfa, S.A. Composition antivieillissement de la peau

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112881333A (zh) * 2021-01-13 2021-06-01 江南大学 一种基于改进免疫遗传算法的近红外光谱波长筛选方法
CN115919673A (zh) * 2022-12-19 2023-04-07 华熙生物科技股份有限公司 一种具有抗氧化作用的组合物及其用途

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