Fragrance compositions with improved sensitization

Abstract

Inventive embodiments disclosed herein include a formulation for decreasing skin sensitizing potency of the fragrance. The formulation includes one or more fragrance oils, ethanol, and PPG-20 methyl glucose ether. The concentration of one or more fragrance oil ranges from 4 to 8 percent by weight, the concentration of the PPG-20 methyl glucose ether modulator ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight.

Description

[0001] FRAGRANCE COMPOSITIONS WITH IMPROVED SENSITIZATION

[0002] CLAIM OF PRIORITY

[0003] This patent application claims the benefit of priority to French Application Serial No. 2214051, filed December 21, 2022, which is incorporated by reference herein in its entirety.

[0004] FIELD OF THE INVENTION

[0005] Inventive subject matter herein relates to the field of perfumery, including compositions and method embodiments for decreasing skin sensitizing potency of the fragrance.

[0006] BACKGROUND OF THE DISCLOSURE

[0007] Experimental and clinical studies have shown that fragrance oils can act as prehaptens or prohaptens by forming allergens that are more potent than a parent substance by activation outside or in the skin. Activation occurs via abiotic, chemical and physical factors, or biotic activation, thus, increasing the risk of sensitization. Some fragrance oils autoxidize on contact with air, forming potent sensitizers that can be an important source for contact allergy to fragrances and fragranced products. Some of them act as prohaptens and are activated in the skin as well.

[0008] SUMMARY

[0009] Inventive embodiments disclosed herein include a formulation for decreasing skin sensitizing potency of the fragrance. The formulation includes one or more fragrance oils, ethanol, and a modulator, also described herein as PPG-20 methyl glucose ether. The concentration of one or more fragrance oils ranges from 4 to 8 percent by weight. The concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight and the concentration of ethanol ranges from 30 to 66 percent by weight. The ratio of fragrance oil to PPG-20 methyl glucose ether ranges from 0.27 to 0.80. The ratio of ethanol- to- PPG-20 methyl glucose ether is within a range of 2.0 to 6.6.

[0010] Inventive embodiments include non-medicinal use of PPG-20 methyl glucose ether for decreasing skin sensitizing potency in a fragrance comprising preparing a fragrance that includes one or more fragrance oils, PPG-20 methyl glucose ether, and ethanol, wherein the ratio of fragrance oils-to-PPG-methyl glucose ether is within a range of either 0.27-to-0.45 or a range of 0;54-to-0.80 and the ratio of ethanol-to-PPG-20 methyl glucose ether is 20-to-4.7 or 5.5-to- 8.0.

[0011] Inventive embodiments include PPG-20 methyl glucose ether, for use in decreasing skin sensitizing potency in a fragrance, comprising preparing a fragrance that includes one or more fragrance oils, PPG-20 methyl glucose ether, and ethanol, wherein the ratio of fragrance oils-to-PPG-20 methyl glucose ether is within a range of either 0.27-to-0.45 or a range of 0.54-to-0.80 and the ratio of ethanol-to-PPG-20 methyl glucose ether is 2.0-to-4.7 or 5.5-to-8.0.

[0012] Inventive embodiments also include a method for decreasing skin sensitizing potency in a fragrance. The method includes preparing a fragrance that includes one or more fragrance oils, PPG-20 methyl glucose ether, and ethanol. The concentration of fragrance oil ranges from 4 to 8 percent by weight. The concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight and the concentration of ethanol ranges from 30 to 66 percent by weight.

[0013] Inventive embodiments also include a method of for decreasing skin sensitizing potency in a fragrance, comprising preparing a fragrance that includes one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of fragrance oil to PPG-20 methyl glucose ether ranges from 0.27 to 0.80.

[0014] Inventive embodiments additionally include a formulation for decreasing skin sensitizing potency of the fragrance, comprising one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of fragrance oil to PPG-20 methyl glucose ether ranges from 0.27 to 0.80.

[0015] Other inventive embodiments include a formulation for decreasing skin sensitizing potency of the fragrance, comprising one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of ethanol- to- PPG- 20 methyl glucose ether is within a range of 2.0 to 5.0. Ethanol concentration ranges from 30 percent to 66 percent by weight.

[0016] Also included is an inventive embodiment for a method for decreasing skin sensitizing potency of the fragrance, comprising preparing a fragrance formulation that includes one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of ethanol- to- PPG-20 methyl glucose ether is within a range of 2.0 to 5.0. Ethanol concentration ranges from 30 percent to 66 percent by weight.

[0017] DESCRIPTION OF THE DRAWINGS:

[0018] FIG. 1 illustrates a summary of steps of the SenzaGen GARDskin dose response method, developed by SenzaGen AB of Lund, Sweden.

[0019] FIG. 2 graphically illustrates application of the GARDskin dose response protocol in a titrated range of concentrations (n=6) starting from a determined top concentration.

[0020] FIG. 3 graphically shows that a formula 1, described herein, showed that a concentration of 15% PPG-20 methyl glucose ether by weight decreased the sensitizing potential of formula 1. Results are summarized in Table 1.

[0021] FIG. 4 graphically shows that a concentration of 10% PPG-20 methyl glucose ether by weight decreased the sensitizing potential for formula 1, described herein. Results are summarized in Table 1.

[0022] FIG. 5 graphically shows that formula 1 concentrations of 5% PPG-20 methyl glucose ether displayed no effect on the sensitizing potential of formula

[0023] 1, described herein. Results are summarized in Table 1.

[0024] FIG. 6 graphically shows that a pentylene glycol, concentration of 15% has no effect on the sensitizing potential of formula. 1, described herein. Results are summarized in Table 1.

[0025] FIG. 7 graphically shows that a concentration of 5% polyurethane-64 has no effect on the sensitizing potential of formula 1, described herein.

[0026] FIG. 8 graphically shows that a concentration of 15% PPG-20 methyl glucose ether significantly decreased the sensitizing potential of a formula 2, described herein with 8% fragrance oil. Results are summarized in Table 1.

[0027] FIG. 9 graphically shows a formula with a concentration of 15% PPG-20 methyl glucose ether displays a decrease of the sensitizing potential of formula

[0028] 2, described herein, with a 4% fragrance oil. The result is not significant, but the effect is visible.

[0029] FIG. 10 graphically shows that a concentration of 15% PPG-20 methyl glucose ether has no effect on the sensitizing potential of formula 2, described herein, with 16% fragrance oil. Results are summarized in Table 1. FIG. 11 graphically shows that the sensitizing potential of formula 2, without a modulator, is equivalent at any fragrance oil doses. Results are summarized in Table 1.

[0030] FIG. 12 graphically shows that a concentration of 15% PPG-20 methyl glucose ether has no effect on the sensitizing potential of formula 2, described herein with 16% fragrance oil.

[0031] DETAILED DESCRIPTION OF THE INVENTION

[0032] Inventive embodiments disclosed herein include a formulation for decreasing skin sensitizing potency of the fragrance. The formulation includes one or more fragrance oils, a modulator such as PPG-20 methyl glucose ether and ethanol. The concentration of fragrance oil ranges from 4 to 8 percent by weight, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight.

[0033] Inventive embodiments also include a method for decreasing skin sensitizing potency in a fragrance. The method includes preparing a fragrance that includes one or more fragrance oils, a PPG-20 methyl glucose ether modulator, and ethanol. The concentration of fragrance oil ranges from 4 to 8 percent by weight, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight.

[0034] A summary of ratios of fragrance oil(s) to PPG-20 methyl glucose ether to ethanol are as follows:

[0035] Range of Ratio of fragrance oil to PPG-20 methyl glucose ether for fragrance oil concentration range 4-8% and PPG methyl glucose ether concentration of 10-15%: 0.27 to 0.80; 4% / 10%=0.4; 5% / 10%=0.5; 6% / 10%=0.6; 7% / 10%=0.7; 8% / 10%=0.8; 4% / l l%=0.36; 5% / l l%=0.45; 6% / l l%=0.54; 7% / l l%=0.63; 8% / l l%=0.72; 4% / 12%=0.33; 5% / 12%=0.41; 6% / 12%=0.50; 7% / 12%=0.58; 8% / 12%=0.67; 4% / 13%=0.31; 5% / 13%=0.38; 6% / 13%=0.46; 7% / 13%=0.54; 8% / 13%=0.61; 4% / 14%=0.29; 5% / 14%=0.36; 6% / 14%=0.43; 7% / 14%=0.50; 8% / 14%=0.57; 4% / 15%=0.27; 5% / l 5%=0.34; 6% / 15%=0.40; 7% / 15%=0.47; 8% / 15%=0.53. Range of Ratio of ethanol to PPG-20 methyl glucose ether for ethanol range 30 to 66 % and PPG-20 methyl glucose ether concentration of 10-15%: 2.0 to 6.6: 30 / 10=3; 0 / 10=4; 50 / 10=5; 55 / 10=5.5; 60 / 10=6.0; 66 / 10=6.6; 30 / 11=2.7;

[0036] 40 / 11=3.6; 50 / 11=4.5; 55 / 11=5.0; 60 / 11=5.4; 66 / 11=6.0; 30 / 12=2.5; 40 / 12=3.3; 50 / 12=4.2; 55 / 12=4.6; 60 / 12=5.0; 66 / 12=5.5; 30 / 13=2.3; 40 / 13=3.1; 50 / 13=3.8; 55 / 13=4.2; 60 / 13=4.6; 66 / 13=5.1; 30 / 14=2.1; 40 / 14=2.8; 50 / 14=3.6; 55 / 14=3.9;

[0037] 60 / 14=4.3; 66 / 14=4.7; 30 / 15=2.0; 40 / 15=2.7; 50 / 15=3.3; 55 / 15=3.7; 60 / 15=4.0; 66 / 15=4.4.

[0038] Definitions

[0039] As used herein, the terms “include”, “includes” and “including” are meant to be non-limiting.

[0040] As used herein, the term “PPG-20 methyl glucose ether” refers to PPG methyl glucose ether polyols. PPG-20 methyl glucose ether is also known as “glucam.” The PPG-20 methyl glucose ether polyols used in accordance with the

[0041] where n has an average value of 20.

[0042] PPG-20 methyl glucose ether is a preferred form of PPG methyl glucose ether employed in formulations and methods disclosed herein.

[0043] Hydrolite 5 green, as used herein has INCI name pentylene glycol,

[0044] FCP as used herein has INCI name polyurethane-64 and trade name, Polyurethane-64.

[0045] As used herein, the term “body splash” means a body care formulation that is applied to the body. Typically, the body splash is applied to the body after bathing and provides a subtle hint of scent to the body. Body splashes are commonly used by consumers who prefer less strong fragrance compositions. A body splash may comprise an ethanol-free composition according to the present invention which comprises from 0.2-8 wt%, relative to the total weight of the composition, of a fragrance component. The body splash may further comprise alkyl polyglucosides as non-ionic surfactants.

[0046] As used herein, the term “body spray” means a formulation comprising fragrance materials intended to be applied to the body to prevent or mask body odor caused by the bacterial breakdown of perspiration on the body (e.g., armpits, feet, and other areas of the body). The body spray may also provide a fragrance expression to the consumers. Typically, body spray compositions are applied as an aerosol spray in an effective amount on the skin of a consumer.

[0047] As used herein, the term “composition” includes a fine fragrance composition intended for application to a body surface, such as for example, skin or hair, e.g., to impart a pleasant odor thereto, or cover a malodour thereof. They are generally in the form of perfume concentrates, perfumes, eau de parfums, eau de toilettes, aftershaves, or colognes. The fine fragrance compositions may be an ethanol-based composition. The term “composition” may also include a cosmetic composition, which comprises a fragrance material for the purposes of delivering a pleasant smell to drive consumer acceptance of the cosmetic composition. The term “composition” may also include body splashes or body sprays. The term “composition” may also include cleaning compositions, such as fabric care composition or home care compositions, including air care compositions (e.g., air fresheners), for use on clothing or other substrates such as hard surfaces (e.g., dishes, floors, countertops). Additional non-limiting examples of “composition” may also include facial or body powder, deodorant, foundation, body / facial oil, mousse, creams (e.g., cold creams), waxes, sunscreens and blocks, bath and shower gels, lip balms, self-tanning compositions, masks and patches.

[0048] As used herein, the term “consumer” means both the user of the composition and the observer nearby or around the user.

[0049] As used herein, the term “fragrance material” and “fragrance materials” relates to a perfume raw material (“PRM”), or a mixture of perfume raw materials (“PRMs”), that are used to impart an overall pleasant odor or fragrance profile to a composition. “Fragrance materials” can encompass any suitable perfume raw materials for fragrance uses, including materials such as, for example, alcohols, aldehydes, ketones, esters, ethers, acetates, nitriles, terpene hydrocarbons, nitrogenous or sulfurous heterocyclic compounds and essential oils. However, naturally occurring plant and animal oils and exudates comprising complex mixtures of various chemical components are also known for use as “fragrance materials”. The individual perfume raw materials which comprise a known natural oil can be found by reference to Journals commonly used by those skilled in the art such as “Perfume and Flavourisf ’ or “Journal of Essential Oil Research”, or listed in reference texts such as the book by S. Arctander, Perfume and Flavor Chemicals, 1969, Montclair, New Jersey, USA and more recently re-published by Allured Publishing Corporation Illinois (1994). Additionally, some perfume raw materials are supplied by the fragrance houses (Firmenich, International Flavors & Fragrances, Givaudan, Symrise) as mixtures in the form of proprietary specialty accords. Non-limiting examples of the fragrance materials useful herein include pro-fragrances such as acetal profragrances, ketal pro-fragrances, ester pro-fragrances, hydrolyzable inorganic- organic pro-fragrances, and mixtures thereof. The fragrance materials may be released from the pro-fragrances in a number of ways. For example, 0 the fragrance may be released as a result of simple hydrolysis, or by a shift in an equilibrium reaction, or by a pH-change, or by enzymatic release.

[0050] As used herein, the term “fragrance profile” means the description of how the fragrance is perceived by the human nose at any moment in time. The fragrance profile may change over time. It is a result of the combination of the low, moderate and high volatile fragrance materials, if present, of a fragrance. A fragrance profile is composed of 2 characteristics: ‘intensity’ and ‘character’. The ‘intensity’ relates to the perceived strength whilst ‘character’ refers to the odor impression or quality of the perfume, e.g., fruity, floral, woody, etc.

[0051] As used herein, the terms “modulator”, and “fragrance modulator” are used interchangeably to designate an agent having the capacity to affect the fragrance profile, such as for example, by impacting the fragrance materials’ evaporation rate. The modulator may mediate its effect by lowering the vapor pressure of the fragrance materials and increasing their adherence to the substrate (skin and / or hair) thus ensuring a less harsh impression of the overdosed fragrance. By incorporating the modulator, it is desired that the fragrance profile, preferably the fragrance components composition attributable to the moderate and low volatile fragrance materials, alone or individually, of the composition can be perceived by a panel of experts or professional evaluators or individual experts or professional evaluators, without the perceived harshness of overdosing (e.g., greater than about 30 wt% of the composition) of the low and moderate fragrance materials is mitigated or absent, as compared to the same perception in the absence of the modulator. Suitable examples of the modulator are provided herein below. However, as discovered by the inventors, simply adding modulators to a traditionally constructed fragrance composition (e.g., classical fragrance pyramid construction without overdose) will only decrease the overall intensity of the fragrance which is undesirable. It is necessary to overdose the key -charactergiving moderate and low volatility materials, above what they would be used at in a traditional fragrance construction, in order to achieve the desired panel of experts or professional evaluators or individual experts or professional evaluators experience.

[0052] As used herein “overdose” can include overdosing a moderate volatile component or high volatile component in aggregate (e.g., greater than 30 wt% of the fragrance component). The term “overdose” can further include overdosing an individual component of the moderate volatile component or the high volatile component (e.g., if the high volatile component includes three oils at least one of the oils may account for a greater wt% of the high volatile component than would be present in a traditional fragrance or a fragrance that is free of the modulators described herein).

[0053] As used herein, the term “substantially non-odorous” means an agent that does not impart an odor of its own when added into a composition of the present invention. For example, a “substantially non-odorous fragrance modulator” does not impart a new odor that alters the character of the fragrance profile of the composition to which it is added. The term “substantially non-odorous” also encompasses an agent that may impart a minimal or slight odor of its own when added into a composition of the present invention. However, the odor imparted by the “substantially non-odorous fragrance modulator” is generally undetectable or tends to not substantively alter the character of the fragrance profile of the composition to which it is added initially or preferably over time. Furthermore, the term “substantially non-odorous” also includes materials that are perceivable only by a minority of people or those materials deemed “anosmic” to the majority of people. Furthermore, the term “substantially non- odorous” also includes materials that may, from particular suppliers, contain an odor due to impurities, such as when the materials contain the impurities at not more than about 5 wt%, preferably not more than 1 wt%, often even not more than 1 part per million (ppm). These impurities maybe removed by purification techniques known in the art as required to make them suitable for use in fragrance compositions of the present invention.

[0054] As used herein, the term “vapor pressure” means the partial pressure in air at a defined temperature (e.g., 25 °C) and standard atmospheric pressure (760 mmHg) for a given chemical species. It defines a chemical species’ desire to be in the gas phase rather than the liquid or solid state. The higher the vapor pressure the greater the proportion of the material that will, at equilibrium, be found in a closed headspace. It is also related to the rate of evaporation of a fragrance material which is defined in an open environment where material is leaving the system. The vapor pressure is determined according to the reference program Advanced Chemistry Development (ACD / Labs) Software Version 14.02, or preferably the latest version update).

[0055] In all embodiments of the present invention, all percentages are by weight of the total composition, as evident by the context, unless specifically stated otherwise. All ratios are weight ratios, unless specifically stated otherwise, and all measurements are made at 25 °C, unless otherwise designated

[0056] As used herein, “skin sensitizing potency” is defined as the relative amount, of chemical per unit area required for the acquisition of skin sensitization in a previously naive individual. For chemical safety assessment, it is of utmost importance to evaluate dose-effect relationships and to classify chemicals according to the strength of their response. Skin sensitizing potency is measured by a GARDskin dose response method. GARDskin is an in vitro test to identify chemical skin sensitizers obtained from SenzaGen, with headquarters in Medicon Village SE-223 81 , Lund, Sweden FIG. 1 illustrates a summary of steps of the GARDskin dose response method. FIG. 2 graphically illustrates application of the GARDskin dose response protocol in a titrated range of concentrations (n=6) starting from a determined top concentration.

[0057] It is understood that the test methods that are disclosed in the Test Methods Section of the present application must be used to determine the respective values of the parameters of Applicants’ inventions as described and claimed herein.

[0058] In all embodiments of the present invention, all percentages are by weight of the total composition, as evident by the context, unless specifically stated otherwise. All ratios are weight ratios, unless specifically stated otherwise, and all measurements are made at 25 °C, unless otherwise designated.

[0059] Compositions

[0060] Disclosed fragrance compositions herein include at least a fragrance component and modulator. The fragrance component includes a wide variety of fragrance materials. The fragrance materials can be grouped in terms of their volatility. Generally, the materials can be grouped as low volatile fragrance materials, moderate volatile fragrance materials, and high volatile fragrance materials. Each group of materials can be associated with various perceptions by a panel of experts or professional evaluators or individual experts or professional evaluators. While not so limited, a high volatile fragrance may be associated with a citrus character; a moderate voile fragrance may be associated with a spicy character; and a low volatile fragrance may be associated with a woody character. Each group of fragrance materials can include synthetic materials or natural materials. The volatility of the fragrance materials can be in reference to an individual fragrance material. Alternatively, in cases where a combination of materials produce a fragrance the volatility may be in reference to that aggregation.

[0061] Decreasing skin sensitizing potency of the fragrance requires that the fragrance composition meets two criteria. First, the concentration of fragrance oil ranges from 4 to 8 percent by weight. Also, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight. Ingredients of the formulation embodiments herein must meet all of these concentration criteria.

[0062] As described herein, the “fragrance materials” have been classified as low, moderate or high volatile fragrance materials according to their volatility by their vapor pressure. This method of classifying fragrance materials by their vapor pressure avoids the problem of different classifications for the same fragrance material according to the traditional approach that relies on their subjective characteristic character. In the case that the fragrance materials are a natural oil, extract or absolute, which comprises a mixture of several compounds, the vapor pressure of the complete oil should be treated a mixture of the individual perfume raw material components using the reference program cited above. The individual components and their level, in any given natural oil or extract, can be determined by direct injection of the oil into a GC-MS column for analysis as known by one skilled in the art. In the scenario that the fragrance materials are a proprietary specialty accord, so called ‘bases’, the vapor pressure, using the reference program cited above, should preferably be obtained from the supplier. However, it is understood by one skilled in the art that they can physically analyze the composition of a full fragrance oil available commercially to identity the fragrance raw materials and their levels using standard GC-MS techniques. This would be irrespective of whether they had been added to the fragrance oil as individual chemicals, as components of naturals or from proprietary bases. Although proprietary bases and naturals are included in our examples, when analyzing a commercially available fragrance via GC-MS one could simply identify the components of the base or natural oil as part of the overall fragrance mixture and their levels, without being able to identify which proprietary base or natural oil the fragrance had come from.

[0063] Fragrance Modulators

[0064] The modulator of interest for the method and formulation embodiments disclosed herein is PPG-20 methyl glucose ether.

[0065] According to various examples, the effect of the substantially non- odorous fragrance modulator on the fragrance profile, particularly the characters of the fragrance profile which is attributable to the low and high volatile fragrance materials, can be improved. By “improved” it is meant that the fragrance profile of the composition, particular the components contributed by at least one of the low and high volatile fragrance materials, can be perceived by the panel of experts or professional evaluators or individual experts or professional evaluators at later time points such as, for example, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 10 hours, and possibly all the way up to 24 hrs after application as compared to controls, e.g., lacking any of the disclosed non-odorous fragrance modulators such as PPG-20 methyl glucose ether.

[0066] By “improved” it can mean that the perception by the panel of experts or professional evaluators or individual experts or professional evaluators, of a harshness of the composition being loaded with greater than 30 wt% of the low- fragrance material is reduced or eliminated.

[0067] In addition to a longevity of a fragrance, users are concerned with skin sensitization potency of cosmetic fragrance formulations. Skin sensitization potency, i.e. Adverse Outcome Pathway, is measured herein with a GARDskin Dose-Response assay, obtained from SenzaGen, headquartered in Lund, Sweden. This assay is a modification of the validated GARDskin protocol , OECD TGP 4.106, that incorporates dose response analysis.

[0068] The GARDskin assay employs a gene expression biomarker to identify skin sensitizers in a human myeloid dendritic-like cell line. The GARDskin Dose-Response investigates the GARDSKIN response values in a titrated range of multiple concentrations in a dose-response manner to find a concentration threshold required to induce a positive decision value. The concentration threshold is used to estimate the inherent sensitizing potency of a chemical. A low value indicates a high inherent skin sensitizing potency and a high value indicates a low inherent skin sensitizing potency. The readout is a cDVO value, describing the lowest concentration required to generate a positive classification. This value correlates with potency and can be used to rank test items by their relative sensitizing potency. This assay allows to evaluate mixtures and delivers continuous potency predictions, crucial point for comparing the modulator’s effect on the formulations’ skin sensitizing potency. One description of the method is described in this article: Quantitative assessment of sensitizing potency using a dose-response adaptation of GARDskin, R. Gradin et al., Scientific Reports, 18904 (2021).

[0069] The potential of the two modulators and one film former to reduce the skin sensitizing potency of fragrance formulations containing different concentrations of fragrance oils was investigated using the GARDskin protocol.

[0070] The GARDskin Dose-Response assay is based on the validated protocols of GARDskin (OECD TG 442E) and incorporates dose-response measurements to derive continuous potency predictions. The readout of the assay corresponds to the lowest concentration required to exceed the binary classification threshold in GARDskin (DV > 0), referred to as the cDVO value. This concentration is inversely correlated to skin sensitizing potency and can be used to predict a corresponding LENA EC3 / human NESIL value in, for instance, micrograms per square centimeter, or be directly interpreted on its own to compare differences in skin sensitizing potency between investigated samples.

[0071] In this study, a total of twelve fragrance formulations were evaluated. The purpose of the study was to determine an efficacious level of PPG-20 methyl glucose ether in fragrance formulations for reducing skin sensitization; assess other fragrance modulators and determine the impact of the fragrance level on PPG-20 methyl glucose ether’s efficacy. Dose-response curves were supplemented with 95% confidence intervals and non-overlapping confidence intervals for cDVO values were used to determine statistically significant differences in potency between investigated samples. The analysis was split in two sample waives using two different fragrance formula chassis (see Table 1 for details):

[0072] Formula 1 compared the effect of two different modulators and one film former, PPG-20 methyl glucose ether, pentylene glycol and polyurethane-64 technology, the film former, on the sensitizing potential of a first fragrance formulation. All samples were compared to a control sample without added modulator. Formula 1 included ethanol in a concentration of 73% and fragrance oil. Concentrations of PPG-20 methyl glucose ether in Formula 1 were 15% w / w, 10% w / w and 5% w / w, respectively. Fragrance oil concentration was held constant at 8% w / w for each test.

[0073] Formula 2 contained the same fragrance oils as Formula 1 but included different concentrations of ethanol and water compared to Formula 1. Formula 2 analyzed the impact of PPG-20 methyl glucose ether on the sensitizing potential of a second type of formulation, designed with different fragrance levels. Each fragrance level had its own negative control. In this sampling for formula 2, PPG-20 methyl glucose ether concentration was 15% w / w. Fragrance oil concentration ranged from 4% to 16%. Ethanol concentration in Formula 2 was 65%.

[0074] Table 1: TESTED SAMPLES:

[0075] GARDskin DOSE RESPONSE METHOD:

[0076] The ability of each sample tested to modulate the sensitization potency was measured using a GARDskin dose response method. The GARDskin dose response method is an in vitro test based on human dendritic-like cell cline, SenzaCell, a set of 200 genes known as the GARD Prediction Signature and a prediction model developed with machine learning technology. A summary of the GARDskin method is shown in FIG. 1. A test sample that includes at least one test substance is prepared and different concentrations of the test sample are grown in cells obtained from SenzaGen AB. A determination is made of the concentration of the test substance where cells react and 90 percent of cells survive. Then, a fresh batch of quality controlled cells are exposed to the determined concentration wherein cells react and 90% survive.

[0077] After exposure to the test sample, total RNA is isolated from SenzaCells. Gene expression is then quantified and analyzed by pattern recognition using a machine learning algorithm based on a fixed set of reference samples. The genes measured are well characterized and are related to several skin sensitization methanistic events. The resulting classification is that of a sensitizer or non-sensitizer. Additional information about the method can be found in a Nature article, “Quantitative assessment of sensitizing potency using a dose-response adaptation of GARDskin,” Nature, article no. 18904 (2021).

[0078] It was hypothesized that the lowest concentration required to exceed the binary classification threshold (DV > 0) and generate a positive classification, referred to as the cDVo concentration, would be informative of sensitizing potency. As such, the proposed dose-response measurements would also be aligned with common toxicological principles for potency assessments, where the response value (DV), the binary threshold (DV > 0), and the derived cDVo concentrations may be viewed as an analogue to the response value (stimulation index (SI)), the binary' threshold (SI > 3), and the EC3 concentrations in the LLNA assay, respectively.

[0079] For FIGs. 2-12 herein, points represent measured response values (decision values) at respective concentrations. Points with white centers represent the GARD input concentrations. Dotted lines describe the log-logistic models, and the solid lines describe the linear interpolations. The running median was used as positive interpolation point to reduce the potenti l impact of noise.

[0080] Application of the GARDskin dose response protocol in a titrated range of concentrations (n=6) starting from a determined top concentration shown in FIG. 2.

[0081] Generation by Support Vector Machine of a Decision Value DV) computing all expression data for each concentration, shown in FIG. 2.

[0082] Estimation of the DV(0) value: lowest concentration required to induce a positive classification (DV>0) shown in FIG. 2.

[0083] Comparison of cDV(O) values of different samples (+ / - modulator), shown in FIG. 2.

[0084] RESULTS:

[0085] FIG. 3 graphically shows that for Formula 1, a concentration of 15% PPG-20 methyl glucose ether (MP22002 shown in FIG. 3), shown in the TESTED SAMPLES table above, by weight decreased the sensitizing potential of Formula 1. A concentration of 15% PPG-20 methyl glucose ether significantly decreased the sensitizing potential of Formula 1. These results confirmed those of a previous GARD study.

[0086] FIG. 4 graphically shows that a concentration of 10% PPG-20 methyl glucose ether by weight (MP22003) also decreased the sensitizing potential for Formula 1. However, Formula 1 concentrations of 5% PPG-20 methyl glucose (MP22004) ether displayed no effect on the sensitizing potential of Formula 1, as shown in FIG. 5. Thus, PPG-20 methyl glucose ether significantly decreased the sensitizing potential of Formula 1 at a dosage between 10% and 15%. The decrease of sensitizing potential was found not to be dose dependent within this range of 10 to 15 percent w / w.

[0087] FIG. 6 shows that a pentylene glycol (MP22005) concentration of 15% had no effect on the sensitizing potential of Formula. 1. Thus, pentyl ene glycol at 15% was not effective in reducing the sensitizing potential of Formula 1. FIG. 7 shows that a concentration of 5% polyurethane-64 had no effect on the sensitizing potential of Formula 1. Thus, polyurethane-64 at 5% (MP22006) was not effective in reducing the sensitizing potential of Formula 1.

[0088] FIG. 8 shows that a concentration of 15% PPG methyl glucose ether significantly decreased the sensitizing potential of a Formula 2 with 8% fragrance oil (MP22008).

[0089] FIG. 9 shows a formula with a concentration of 15% PPG-20 methyl glucose ether displayed a decrease of the sensitizing potential of Formula 2, with a 4% fragrance oil (MP22011). The result was not significant, but the effect is visible.

[0090] FIG. 10 showed that a concentration of 15% PPG-20 methyl glucose ether had no effect on the sensitizing potential of Formula 2 with 16% fragrance oil (MP22010). Thus, the reducing effect of 15% PPG-20 methyl glucose ether on the sensitizing potential of Formula 2 is visible at fragrance oil levels between 4% and 8%.

[0091] FIG. 11 shows that the sensitizing potential of Formula 2, without a modulator, is equivalent at any fragrance oil doses. Testing was performed for formula 2 with 4% fragrance oil (MP22012), 8% fragrance oil (MP22007) and 16% fragrance oil (MP22010).

[0092] FIG. 12 shows that a concentration of 15% PPG-20 methyl glucose ether had no effect on the sensitizing potential of Formula 2 with 16% fragrance oil (MP22009). The effect of 15% PPG-20 methyl glucose ether on Formula 2 with either 4% (MP22011) or 8% (MP22008) fragrance oil was similar. Thus, the reducing effect of 15% PPG-20 methyl glucose ether on the sensitizing potential of formula 2, visible between 4% and 8% fragrance oil, was not dose dependent.

[0093] While in the foregoing specification this invention has been described in relation to certain preferred embodiments thereof, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein may be varied considerably without departing from the basic principles of the invention.

Claims

What is claimed is:

1. A formulation for decreasing skin sensitizing potency of the fragrance, comprising one or more fragrance oils, PPG-20 methyl glucose ether modulator and ethanol, wherein the concentration of fragrance oil ranges from 4 to 8 percent by weight, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight.

2. A method for decreasing skin sensitizing potency in a fragrance, comprising preparing a fragrance that includes one or more fragrance oils, PPG- 20 methyl glucose ether modulator, and ethanol, wherein the concentration of fragrance oil ranges from 4 to 8 percent by weight, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight.

3. The method of claim 2, wherein the PPG-20 methyl glucose ether added to the fragrance is a mixture of alpha and beta glucoside.

4. A method of for decreasing skin sensitizing potency in a fragrance, comprising preparing a fragrance that includes one or more fragrance oils, PPG- 20 methyl glucose ether and ethanol, wherein the ratio of one or more fragrance oils to PPG-20 methyl glucose ether ranges from 0.27 to 0.80 and the ethanol concentration ranges from 30 to 66 percent by weight.

5. A formulation for decreasing skin sensitizing potency of the fragrance, comprising one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of fragrance oil to PPG-20 methyl glucose ether ranges from 0.27 to 0.80 and the ethanol concentration ranges from 30 to 66 percent by weight.

6. A formulation for decreasing skin sensitizing potency of the fragrance, comprising one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of ethanol- to- PPG-20 methyl glucose ether is withina range of 2.0 to 5.0 and the concentration of ethanol ranges from 30 to 66 percent by weight.

7. A method for decreasing skin sensitizing potency of the fragrance, comprising preparing a fragrance formulation that includes one or more fragrance oils, PPG-20 methyl glucose ether and ethanol, wherein the ratio of ethanol- to- PPG-20 methyl glucose ether is within a range of 2.0 to 6.6 and the concentration of ethanol ranges from 30 to 60 percent by weight.

8. A formulation for decreasing skin sensitizing potency of the fragrance, comprising one or more fragrance oils, PPG-20 methyl glucose ether modulator and ethanol, wherein the concentration of fragrance oil ranges from 4 to 8 percent by weight, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight; and wherein the ratio of fragrance oil to PPG-20 methyl glucose ether ranges from 0.27 to 0.80 and the ratio of ethanol to PPG-20 glucose ether ranges from 2.0 to 6.6.

9. A method for decreasing skin sensitizing potency in a fragrance, comprising preparing a fragrance that includes one or more fragrance oils, PPG- 20 methyl glucose ether modulator, and ethanol, wherein the concentration of fragrance oil ranges from 4 to 8 percent by weight, the concentration of PPG-20 methyl glucose ether ranges from 10 to 15 percent by weight, and the concentration of ethanol ranges from 30 to 66 percent by weight; and wherein the ratio of fragrance oil to PPG-20 methyl glucose ether ranges from 0.27 to 0.80 and the ratio of ethanol to PPG-20 glucose ether ranges from 2.0 to 6.6.

10. Inventive embodiments include non-medicinal use of PPG-20 methyl glucose ether for decreasing skin sensitizing potency in a fragrance comprising preparing a fragrance that includes one or more fragrance oils, PPG-20 methyl glucose ether, and ethanol, wherein the ratio of fragrance oils-to-PPG-20 methyl glucose ether is within a range of either 0.27-to-0.45 or a range of 0;54-to-0.80 and the ratio of ethanol-to-PPG-20 methyl glucose ether is 20-to-4.7 or 5.5-to- 8.0.

11. PPG-20 methyl glucose ether for use in decreasing skin sensitizing potency in a fragrance, comprising preparing a fragrance that includes one or more fragrance oils, PPG-20 methyl glucose ether, and ethanol, wherein the ratio of fragrance oils-to-PPG-20 methyl glucose ether is within a range of ether 0.27-to-0.45 or a range of 0.54-to-0.80 and the ratio of ethanol-to PPG-20 methyl glucose ether is 2.0-to-4.7 or 5.5 to 8.0.

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