A compound, a probe, and methods thereof

Abstract

The present disclosure provides a compound of Formula (I), wherein A is an aryl ring comprising C10 to C20 atoms; R1 and R2 are independently selected from C1-6 alkyl, or C6-12 aryl; or R1 and R2 combine together to form heterocyclyl ring having 3 to 8 atoms; m is in a range of 0 to 10; and n is in a range of 0 to 10. The present disclosure further provides a probe comprising the compound of Formula (I) for detecting a sensitizer. Furthermore, the present disclosure provides a method of detecting a sensitizer.

Description

A COMPOUND, A PROBE, AND METHODS THEREOFFIELD OF INVENTION

[0001] The present disclosure relates, in general, to analysis, and safety assessment of skin-reactive compounds or sensitizers. In particular, the present disclosure relates to a compound, a probe, and a method of detecting skin-sensitizers.BACKGROUND OF INVENTION

[0002] Skin sensitization is an important endpoint for cosmetics risk assessment considering both the nature of the used ingredients and the post-marketing surveillance. The necessary regulatory measures for the prevention of induction and elicitation of allergies are addressed in several regulatory demands and in common market practices. The coexistence of regulatory needs and recent developments in skin sensitization testing protocols for chemicals, fragrance materials, and cosmetics allows for proper hazard and potency estimation, and risk management without the usage of in vivo methods with ethical concerns. Owing to the complexity of the biological mechanisms associated with skin sensitization, integrated approaches combining different chemical, biological, and in silico methods are recommended to replace conventional animal tests.

[0003] Chemical methods are intended to characterize the potential of a sensitizer to induce earlier molecular initiating events. The presence of an electrophilic mechanistic domain is considered one of the essential chemical features to covalently bind to the biological target and induce further haptenation processes. Current in chemico assays rely on the quantification of unreacted model nucleophiles after incubation with the candidate sensitizer.

[0004] Compounds implicated in causing Allergic Contact Dermatitis (ACD) are generally electrophilic in nature. The existing techniques or assays utilize the formation of covalent bonding between electrophiles and nucleophiles due to the interaction of skin sensitizers with proteins, peptides, and nucleophiles representing proteins or peptides. However, these techniques incur longer incubation time and also do not identify pro-electrophiles.

[0005] Therefore, there is a need to develop a reactive compound for use as a probe and a method to identify potential electrophilic skin-sensitizers rapidly and to differentiate an electrophilic sensitizer and a pro-electrophilic sensitizer.SUMMARY OF THE INVENTION

[0006] In an aspect of the present disclosure, there is provided a compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereofFormula (I) wherein A is an aryl ring comprising Cio to C20 atoms;Ri and R2 are independently selected from Ci-6 alkyl, or C6-12 aryl; or Ri and R2 combine together to form heterocyclyl ring having 3 to 8 atoms; and m is in a range of 0 to 10; and n is in a range of 0 to 10.

[0007] In another aspect of the present disclosure, there is provided a probe for detecting a sensitizer comprising the compound of Formula (I) as disclosed herein.

[0008] In a further aspect of the present disclosure, there is provided a method of detecting a sensitizer, the method comprising: (a) mixing the compound of Formula (I) or the probe, as disclosed herein with the sensitizer in the presence of a base to obtain a first solution; (b) incubating the first solution for a time period of 0.5 to 2 hours to obtain an incubated solution; (c) spectral analysis of the incubated solution to measure the compound of Formula (I); and (d) identifying the sensitizer.

[0009] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essentialfeatures of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.BRIEF DESCRIPTION OF THE FIGURES

[0010] The following drawings form a part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.

[0011] Figure 1 depicts the chromatograms of incubated solutions of compound of Formula (I) with (a) p-benzoquinone, (b) 2-methyl 4-isothiazolin 3-one, and (c) famesal, in accordance with an embodiment of the present disclosure.

[0012] Figure 2 depicts the chromatograms of the incubated solutions of p- benzoquinone with (a) compound of Formula (I) (NNDNAC); and (b) N-(2-(l- naphthyl)acetyl)-L-cysteine (NAC-ADRA), at varying incubation time 1 hr and 24 hours, in accordance with an embodiment of the present disclosure.

[0013] Figure 3 depicts the chromatograms of incubated solutions of compound of Formula (I) with a concoction having (a) sensitizer as p-benzoquinone, and (b) nonsensitizer as dimethyl isophthalate, in accordance with an embodiment of the present disclosure.DESCRIPTION OF THE INVENTION

[0014] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features. Definitions

[0015] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. Thesedefinitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.

[0016] The articles “a”, “an” and “the” are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

[0017] The terms “comprise” and “comprising” are used in the inclusive, open sense, meaning that additional elements may be included. It is not intended to be construed as “consists of only”.

[0018] The term “at least one” is used to mean one or more and thus includes individual components as well as mixtures / combinations.

[0019] Throughout this specification, unless the context requires otherwise the word “comprise”, and variations such as, “comprises” and “comprising”, will be understood to imply the inclusion of a stated element or step or group of element or steps but not the exclusion of any other element or step or group of element or steps.

[0020] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably.

[0021] The meaning of various terms used in the description shall now be illustrated.

[0022] According to an embodiment herein, the term “Ci-6 alkyl” refers to straightchain or branched Ci to C , alkyl which may be optionally substituted. Representative examples of alkyl include methyl, ethyl, propyl, 1 -methylethyl, butyl, 1 -methylpropyl, 2-m ethylpropyl, 1,1 -dimethylethyl, pentyl, 1 -methylbutyl, 2-methylbutyl, 3 -methylbutyl, 2,2-dimethylpropyl, 1 -ethylpropyl, hexyl, 1,1- dimethylpropyl, 1,2-dimethylpropyl, 1 -methylpentyl, 2-methylpentyl, 3- methylpentyl, 4-methylpentyl, 1,1 -dimethylbutyl, 1,2-dimethylbutyl, 1,3- dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3, 3 -dimethylbutyl, 1- ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 1 -ethyl- 1- methylpropyl and l-ethyl-2-methylpropyl or the different isomers.

[0023] According to an embodiment herein, the term “aryl” refers to the cyclic or polycyclic (spiro, fused, bridged, non-fused) ring groups that satisfy Huckel’s rule. Representative examples of aryl rings include benzene, naphthalene, chrysene, pyrene, anthracene, phenanthrene, or the like.

[0024] According to an embodiment herein, the term “heterocycle” or “heterocyclic” or “heterocyclyl” includes “aromatic heterocycle”, “nonaromatic heterocycle” or polycyclic or bicyclic (spiro, fused, bridged, non-fused) ring compounds in which the ring may be aromatic or non-aromatic, wherein the heterocycle ring contains at least one heteroatom selected from nitrogen, oxygen, and sulphur along with one or more carbon atoms, with a total of three to eight atoms.

[0025] The term “solvates”, as used herein, refers to a compound wherein its crystal lattice contains one or more solvent molecule. Non-limiting examples of solvates include hydrates when solvent is water, ammoniates when the solvent is ammonia, alcoholates when the solvent is an alcohol, and etherates when the solvent is an ether.

[0026] The term “salt” as used herein, refers to salts of acid or base of compound of Formula (I). Non-limiting examples of acidic salts include both inorganic acidic salts, such as hydrochloric, sulphuric, phosphoric, diphosphoric, hydrobromic, hydroiodic and nitric acid, and organic acidic salts, such as citric, fumaric, maleic, malic, mandelic, ascorbic, oxalic, succinic, tartaric, benzoic, acetic, methane sulphonic, ethane sulphonic, benzene sulphonic or p-toluenesulphonic acid. Nonlimiting examples of basic salts include alkali metal salts of sodium or potassium hydroxides and alkali earth metal salts of calcium or magnesium hydroxides and organic bases, for example alkyl amines, arylalkyl amines and heterocyclic amines.

[0027] The compounds of Formula (I) described herein may contain one or more chiral centers and / or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers), regioisomers, enantiomers or diastereomers. Accordingly, the chemical structures depicted herein encompass all possible enantiomers and stereoisomers of the illustrated or identified compounds including the stereoisomerically pure form (e.g., geometrically pure,enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures. Enantiomeric and stereoisomeric mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the person skilled in the art. The compounds may also exist in several tautomeric forms including the enol form, the keto form, and mixtures thereof. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated or identified compounds. It is also understood that some isomeric form such as diastereomers, enantiomers and geometrical isomers can be separated by physical and / or chemical methods and by those skilled in the art. Further the compounds can exist as mixture having each of the enantiomer in equal amount to form racemates or racemic mixture.

[0028] According to an embodiment herein, the term “sensitizer(s)” or “skin- sensitizer(s)” or “skin-sensitizing compound(s)” refer(s) to substances that are reactive when applied on a skin and may cause damage on the surface of the skin upon contact, owing to the extent of its reactivity.

[0029] According to an embodiment herein, the term “electrophile” refers to a compound which tends to attract or acquire electrons and are thereby reactive in nature. For the purpose of the present disclosure, the term “electrophile” refers to “hapten” which are immunogenic compounds that are electrophilic in nature. These haptens cause allergic reaction upon contact with skin. For the purpose of the disclosure, electrophiles are electrophilic skin-sensitizing compounds and are also referred to as haptens.

[0030] According to an embodiment herein, the term “pro-electrophile” refers to a compound which upon being subjected to certain external conditions, gets converted to an electrophile. For the purpose of the present disclosure, the term “pre-hapten” refers to the immunogenic compounds that are non-reactive in nature which gets converted to hapten upon being subjected to certain external conditions. Pre-haptens upon conversion to haptens cause allergic reactions on skin. For the purpose of the present disclosure, pro-electrophiles are pro-electrophilic skinsensitizing compounds and are also referred to as pre-haptens.

[0031] All percentages, parts and ratios are based upon the total weight of the compositions of the present disclosure unless otherwise indicated. Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a time period range of about 0.5 to 2 hours, should be interpreted to include not only the explicitly recited limits of about 0.5 to about 2 hours, but also to include sub-ranges, such as 0.5 to 0.75 hours, 1.25 to 1.75 hours and so forth, as well as individual amounts, including fractional amounts, within the specified ranges, such as 0.5 h, 1.0 h, 1.25 h, and 1.75 h, for example.

[0032] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference.

[0033] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally equivalent products, and methods are clearly within the scope of the disclosure, as described herein.

[0034] The present disclosure provides a compound of Formula (I), a probe comprising the compound of Formula (I) and a method of detecting a sensitizer utilizing the compound of Formula (I). The present disclosure provides an efficient in-chemico characterization tool to identify the potential skin- sensitizers with fast and high throughput. The probe and the method of detecting a sensitizer of the present disclosure are suitable and provide inexpensive approaches for prescreening large chemical compounds. The method of detecting a sensitizer using the probe comprising the compound of Formula (I) is advantageous over theconventional time consuming and costly in vitro and in vivo evaluations of the skinsensitizing compounds. Furthermore, the method disclosed herein facilitates to distinguish pro-electrophile sensitizers from electrophile sensitizers which the existing methods fails to.

[0035] Embodiments herein provide a compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof,Formula (I) wherein A is an aryl ring comprising Cio to C20 atoms;Ri and R2 are independently selected from Ci-6 alkyl, or C6-12 aryl; or Ri and R2 combine together to form heterocyclyl ring having 3 to 8 atoms; m is in a range of 0 to 10; and n is in a range of 0 to 10.

[0036] The compound of Formula (I) of the present invention makes it possible in particular for detecting a sensitizer with fast throughput and by an economic approach. Embodiments herein also provide a probe comprising a compound of Formula (I).

[0037] Embodiments herein also provide a method of detecting a sensitizer and to categorize a pro-electrophile and an electrophile.Compound

[0038] Embodiments herein provide a compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof, according to the present invention,Formula (I) wherein A is an aryl ring comprising Cio to C20 atoms;Ri and R2 are independently selected from Ci-6 alkyl, or C6-12 aryl; or Ri and R2 combine together to form heterocyclyl ring having 3 to 8 atoms; m is in a range of 0 to 10; and n is in a range of 0 to 10.

[0039] According to an embodiment herein, it makes it possible, in particular, to efficiently detect a sensitizer using the compound of Formula (I).

[0040] According to a particular embodiment herein, A is an aryl ring comprising Cio to Cis atoms; Ri and R2 are independently selected from C1-6 alkyl; m is in a range of 0 to 3; and n is in a range of 0 to 3.

[0041] According to a particular embodiment herein, A is selected from naphthalene, anthracene, chrysene, phenanthrene, or pyrene; Ri and R2 are independently selected from C1-6 alkyl; m is in a range of 0 to 3; and n is in a range of 0 to 3.

[0042] According to a more particular embodiment herein, A is naphthalene, Ri and R2 are independently Ci alkyl; m is 1; and n is 0.

[0043] According to an embodiment herein, the compound of Formula (I) acts as a nucleophile to react with reactive electrophilic or pro-electrophilic sensitizers and thereby detects reactive species. The compound provides specific detection of proelectrophile sensitizers in a short incubation time period of 0.5 to 2 hours. Moreover, the compound of Formula (I), according to embodiments herein, employed in a reactivity probe overcomes the drawbacks associated with the time consumption, difficulty in method of detection, cost of reagents, and high throughput.

[0044] According to other embodiments herein, there is provided a use of the compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof, for the detection of skin- sensitizing compounds. The compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof,according to further embodiments herein, is used in a probe for efficient, fast, and convenient detection of sensitizers.

[0045] According to other embodiments herein, there is provided a use of the compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof, for detecting a sensitizer in a concoction. The compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof, according to further embodiments herein, is used in a probe for efficient, fast, and convenient detection of sensitizers.Probe

[0046] Embodiments herein provide a probe for detecting a sensitizer comprising the compound of Formula (I), according to the present invention.

[0047] According to an embodiment herein, the term “probe” refers to a system for analysing the property of a target by reversibly binding to and altering the function of the target. According to another embodiment herein, the probe is a molecular probe used for detecting sensitizers by reacting with them. The sensitizers, according to embodiments herein, is an electrophile or a pro-electrophile.

[0048] According to other embodiments herein, the sensitizer is an electrophilic or pro-electrophilic skin- sensitizing compound.

[0049] According to other embodiments herein, the probe comprises a compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereof, according to the present invention,Formula (I) wherein A is an aryl ring comprising Cio to C20 atoms;Ri and R2 are independently selected from C1-6 alkyl, or C6-12 aryl; or Ri and R2 combine together to form heterocyclyl ring having 3 to 8 atoms; m is in a range of 0 to 10; and n is in a range of 0 to 10.

[0050] According to a particular embodiment herein, the probe comprises a compound of Formula (I), wherein A is an aryl ring comprising Cioto Cis atoms; Ri and R2 are independently selected from C1-6 alkyl; m is in a range of 0 to 3; and n is in a range of 0 to 3.

[0051] According to a particular embodiment herein, the probe comprises a compound of Formula (I), wherein A is selected from naphthalene, anthracene, chrysene, phenanthrene, or pyrene; Ri and R2 are independently selected from C1-6 alkyl; m is in a range of 0 to 3; and n is in a range of 0 to 3.

[0052] According to other embodiments herein, the probe comprises a compound of Formula (I), wherein A is naphthalene, Ri and R2 are independently Ci alkyl; m is 1; and n is 0.

[0053] According to other embodiments herein, the probe comprises a compound of Formula (I) with other non-reactive components which facilitate detection of skin-sensitizing compounds. The non-reactive components are inert components, in general, that may be employed during the formulation of the probe.

[0054] According to further embodiments herein, there is disclosed a use of the probe comprising the compound of Formula (I) for detecting a sensitizer. The probe exhibits rapid and effective detection of electrophilic and pro -electrophilic sensitizers. The probe, according to embodiments herein detects a sensitizer by a method as disclosed herein.Method

[0055] Embodiments herein include a method of detecting sensitizers. The method includes detecting of sensitizers, particularly electrophilic and pro- electrophilic sensitizers.

[0056] Embodiments herein provide a method of detecting a sensitizer using the compound of Formula (I) as disclosed herein. According to an embodiment herein, the method of detecting a sensitizer comprises: (a) mixing the compound of Formula (I) with the sensitizer in the presence of a base to obtain a first solution; (b) incubating the first solution for a time period of 0.5 to 2 hours to obtain an incubated solution; (c) spectral analysis of the incubated solution to measure the compound of Formula (I); and (d) identifying the sensitizer.

[0057] Embodiments herein provide a method of detecting a sensitizer using the compound of Formula (I) as disclosed herein. According to an embodiment herein, the method of detecting a sensitizer comprises: (a) mixing the compound of Formula (I), wherein A is naphthalene, Ri and R2 are independently Ci alkyl; m is 1; and n is 0, with the sensitizer in the presence of a base to obtain a first solution; (b) incubating the first solution for a time period of 0.5 to 2 hours to obtain an incubated solution; (c) spectral analysis of the incubated solution to measure the compound of Formula (I); and (d) identifying the sensitizer.

[0058] According to an embodiment herein, the compound of Formula (I) as disclosed herein or the probe as disclosed herein, is in a concentration of at least 5pM, with respect to the total volume of the first solution. According to a particular embodiment herein, the compound of Formula (I) as disclosed herein or the probe as disclosed herein, is in a concentration range of 5 to 50 pM, with respect to the total volume of the first solution.

[0059] According to an embodiment herein, the sensitizer, is in a concentration of at least ImM, with respect to the total volume of the first solution. According to a particular embodiment herein, the sensitizer is in a concentration range of 1 to 10 mM, with respect to the total volume of the first solution.

[0060] Embodiments herein provide a method of detecting a sensitizer in a concoction using the compound of Formula (I) as disclosed herein. According to an embodiment herein, the method of detecting a sensitizer in a concoction, the method comprises: (a) mixing the compound of Formula (I), with the concoction in thepresence of a base to obtain a mixture; (b) incubating the mixture for a time period of 0.5 to 2 hours to obtain an incubated mixture; (c) subjecting the incubated mixture to spectral analysis to detect the presence of the sensitizer.

[0061] Embodiments herein provide a method of detecting a sensitizer in a concoction using the compound of Formula (I) as disclosed herein. According to an embodiment herein, the method of detecting a sensitizer in a concoction, the method comprises: (a) mixing the compound of Formula (I), wherein A is naphthalene, Ri and R2 are independently Ci alkyl; m is 1; and n is 0, with the concoction in the presence of a base to obtain a mixture; (b) incubating the mixture for a time period of 0.5 to 2 hours to obtain an incubated mixture; (c) subjecting the incubated mixture to spectral analysis to detect the presence of the sensitizer

[0062] According to an embodiment herein, the concoction comprising a formulation and a sensitizer, and the formulation comprises at least one active material and at least two additives.

[0063] According to an embodiment herein, the term “concoction” refers to the mixture of a formulation and a sensitizer, wherein the formulation is a blend of at least one active material and at least two additives, and wherein the sensitizer is an electrophile or a pro -electrophile.

[0064] According to an embodiment herein, the term “method of detecting a sensitizer in a concoction” refers to a method by which a concoction is analysed or checked for the presence of a sensitizer using the compound of Formula (I) as disclosed herein or the probe as disclosed herein.

[0065] According to an embodiment herein, the incubated mixture is subjected to spectral analysis for identifying the presence of sensitizer by measuring the depletion of the compound of Formula (I).

[0066] According to an embodiment herein, the compound of Formula (I) as disclosed herein or the probe as disclosed herein, is in a concentration of at least 5pM, with respect to the total volume of the mixture. According to a particular embodiment herein, the compound of Formula (I) as disclosed herein or the probeas disclosed herein, is in a concentration range of 5 to 50 |iM, with respect to the total volume of the mixture.

[0067] According to another embodiment herein, the sensitizer is in a concentration of at least ImM, with respect to the total volume of the mixture. According to a particular embodiment herein, the compound of Formula (I) as disclosed herein or the probe as disclosed herein, is in a concentration range of 1 to 5 mM, with respect to the total volume of the mixture.

[0068] According to an embodiment herein, the formulation is in a concentration range of 0.1 to 2 mg / mL, with respect to the total volume of the mixture. According to a particular embodiment herein, the formulation is in a concentration range of 0.3 to 1 mg / mL, with respect to the total volume of the mixture. According to a more particular embodiment herein, the formulation is in a concentration of 0.5 mg / mL, with respect to the total volume of the mixture.

[0069] According to an embodiment herein, the at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or combinations thereof; and at least two additives are selected from a surfactant, a fatty substance, a solvent, a polymer, a preservative, or combinations thereof. According to a particular embodiment herein, the at least one active material is a combination of trisodium ethylenediamine disuccinate and triethanolamine.

[0070] According to another embodiment herein, the surfactant is selected from C15-20 alkyl esters, C15-20 alkenyl esters, C15-20 alkyl-C2-io alkanol esters or combinations thereof; the fatty substance is selected from C15-20 alcohols; the solvent is selected from water, C5-10 alkane diols, C15-20 alkanes, or combinations thereof; the polymer is selected from C2-10 polyalkenes, carbomers or combinations thereof; and the preservative is selected from Ce-io aryl alcohols, Ce-io aryloxy alkanols, or combinations thereof. According to another particular embodiment herein, the surfactant is selected from glyceryl stearate, polyethylene glycol- 100 stearate, polyethylene glycol-40 stearate, or combinations thereof; the fatty substance is selected from cetyl alcohol, stearyl alcohol, or combinations thereof;the solvent is selected from water, octane- 1,2-diol, isohexadecane, or combinations thereof; the polymer is selected from hydrogenated polyisobutene, copolymer of acrylic acid and ethyl acetate / cyclohexane, or combinations thereof; and the preservative is 2-phenoxymethanol.

[0071] According to an embodiment herein, the base is selected from 1,8- diazabicy clo [5.4.0] undec-7 -ene(DB U) , 1,5 -diazabicyclo [4.3.0] non-5 -ene(DBN) , l,4-diazabicyclo[2.2.2]octane (TED), 7-methyl-l,5,7-triazabicyclo[4.4.0] dec-5- ene (MTBD), l,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD), 4-dimethylamino pyridine, pyridine, piperazine or combinations thereof. According to a particular embodiment herein, the base is 1,8-diazabicyclo [5.4.0]undec-7-ene. According to another particular embodiment herein, the base is l,5-diazabicyclo[4.3.0]non-5- ene.

[0072] According to another embodiment herein, incubating the first solution is carried out for a time period of 0.5 to 2 hours. According to a particular embodiment herein, incubating the first solution is carried out for a time period of 0.75 to 1.5 hours. According to a more particular embodiment herein, incubating the first solution is carried out for a time period of 1 hour.

[0073] According to more embodiments herein, identifying the sensitizer involves measuring depletion of the compound of Formula (I). According to other embodiments herein, the compound of Formula (I) being a nucleophile reacts with an electrophilic sensitizer, leads to depletion of the compound of Formula (I).

[0074] According to a further embodiment herein, the spectral analysis of the incubated solution is carried out by chromatographic and fluorescence detection. According to a particular embodiment herein, the spectral analysis is carried out by liquid chromatography followed by fluorescence detection. According to a more particular embodiment herein, the spectral analysis is carried out by liquid chromatography followed by fluorescence detection, to measure the presence of the compound of Formula(I) in the incubated solution and correlate todepletion of the compound of Formula (I). The depletion of the compound of Formula (I) categorizes the sensitizer as an electrophile or a pro-electrophile.

[0075] According to yet other embodiments herein, there is provided a use of the method of detecting a sensitizer as disclosed herein for the pre-screening of a cosmetic composition. The method as disclosed herein is used as an analytical assessment technique of a composition to understand the reactivity of the constituent compounds which result in skin-sensitization.Composition

[0076] Embodiments herein include a composition comprising the compound of Formula (I) as disclosed herein or the probe as disclosed herein. The composition comprises a concoction, wherein the formulation comprises at least one active material, at least two additives and at least one sensitizer.

[0077] Embodiments herein provide a composition comprising the compound of Formula (I) as disclosed herein or the probe as disclosed herein and a concoction. According to an embodiment herein, the composition comprises: (a) the compound of Formula (I) as disclosed herein or the probe as disclosed herein; and (b) a concoction, wherein the concoction comprises at least one active material, at least two additives and at least one sensitizer.

[0078] According to an embodiment herein, the at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or combinations thereof; and at least two additives are selected from a surfactant, a fatty substance, a solvent, a polymer, a preservative, or combinations thereof.

[0079] According to another particular embodiment herein, the at least one active material is a combination of trisodium ethylenediamine disuccinate, and triethanolamine.

[0080] According to yet another embodiment herein, the surfactant is selected from C15-20 alkyl esters, C15-20 alkenyl esters, C15-20 alkyl-C2-io alkanol esters or combinations thereof; the fatty substance is selected from C15-20 alcohols;the solvent is selected from water, C5-10 alkane diols, C 15-20 alkanes, or combinations thereof; the polymer is selected from C2-10 polyalkenes, carbomers or combinations thereof; and the preservative is selected from Ce-io aryl alcohols, Ce-io aryloxy alkanols, or combinations thereof. According to another particular embodiment herein, the surfactant is selected from glyceryl stearate, polyethylene glycol- 100 stearate, polyethylene glycol-40 stearate, or combinations thereof; the fatty substance is selected from cetyl alcohol, stearyl alcohol, or combinations thereof; the solvent is selected from water, octane- 1,2-diol, isohexadecane, or combinations thereof; the polymer is selected from hydrogenated polyisobutene, copolymer of acrylic acid and ethyl acetate / cyclohexane, or combinations thereof; and the preservative is 2-phenoxymethanol.

[0081] According to an embodiment herein, at least one sensitizer is selected from an electrophilic sensitizer, a pro-electrophilic sensitizers, or combinations thereof.Use

[0082] Embodiments herein include a use of the compound of Formula (I) as disclosed herein for detecting a sensitizer.

[0083] According to an embodiment herein, the compound of Formula (I), may be used for detecting a sensitizer. The compound may be used for detecting and categorising a sensitizer into an electrophile or a pro-electrophile. The compound exhibits quick and accurate detection and identification of electrophilic and pro- electrophilic sensitizers. The compound, according to embodiments herein detects a sensitizer by a method as disclosed herein.

[0084] According to another embodiment herein, the compound of Formula (I), may be used for detecting a sensitizer by mixing the compound with the sensitizer in the presence of a base to obtain a first solution; incubating the first solution for a time period of 0.5 to 2 hours to obtain an incubated solution; spectral analysis of the incubated solution to measure the compound of Formula (I); and identifying the presence of sensitizer.

[0085] According to another embodiment herein, the compound may more particularly be a compound for detecting and identifying a sensitizer, particularly for qualitative identification if a compound is a sensitizer or a non-sensitizer, and for qualitative identification if a sensitizer is an electrophile, or a pro-electrophile. Preferably, the compound, according to embodiments herein, is a compound used for detecting all types of sensitizers, electrophile or pro-electrophile. Accordingly, the compound, in various embodiments herein, may be a probe or a composition.

[0086] According to yet another embodiment herein, the compound may be used to detect and categorise a sensitizer in a concoction. The concoction comprises a formulation comprising at least one active material and at least two additives, and a sensitizer. The concoction may be in any suitable form, for example, gel, liquid gel, or liquid form. The concoction may further be a cosmetic product which is applied directly on to skin.

[0087] According to still another embodiment herein, the compound may be used to detect a sensitizer at ambient temperatures within a shorter incubation time period in a range of 0.5 to 2 hours.

[0088] Embodiments herein further include a use of the probe comprising the compound of Formula (I) for detecting a sensitizer. The probe exhibits rapid and effective detection of electrophilic and pro-electrophilic sensitizers. The probe, according to embodiments herein detects a sensitizer by a method as disclosed herein.

[0089] According to an embodiment herein, the probe comprising the compound of Formula (I), may be used for detecting sensitizer in a concoction.

[0090] According to yet another embodiment herein, there is provided a use of the method of detecting a sensitizer as disclosed herein for the pre-screening of a compound to be used in the manufacture of a cosmetic composition. The method as disclosed herein is used as an analytical assessment technique to understand the reactivity of the specific compounds which result in skin-sensitization. The methodas disclosed herein is used for qualitative detection of a sensitizer and qualitative identification if a sensitizer is an electrophile, a pro-electrophile, or a non- sensitizer.

[0091] According to yet another embodiment herein, there is provided a use of the method of detecting a sensitizer in a concoction as disclosed herein, for the prescreening of a cosmetic composition.

[0092] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, it is understood that other implementations are possible and included within the scope of the present invention.EXAMPLES

[0093] The disclosure will now be illustrated with following examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices, and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply.Example 1Compound of Formula (I)

[0094] The compound of Formula (I) of the present disclosure, N-N dimethyl N-(2-(l-naphthyl)acetyl)-L-cysteine (NNDNAC) having A as naphthalene, Ri and R2 independently being Ci alkyl; m as 1; and n as 0, was prepared by the method described herein.Formula (I)Preparation of the compound of Formula (I)

[0095] The compound of Formula (I) was prepared by the process as illustrated inScheme 1.Scheme 1Step 1: 5-bromonaphthalen-l-amine (2):

[0096] To a stirred solution of l-bromo-5-nitronaphthalene (1), (20.0 g, 79.36 mmol, 1.0 eq.) in aqueous methanol (200 mL), Fe (22.16 g, 396.82 mmol, 5.0 eq.) followed by ammonium chloride (42.44 g, 793.6 mmol, 10 eq.) were added at room temperature to obtain a reaction mixture. The reaction mixture was stirred at 75 °C for 1 h to obtain a reaction mass. Progress of the reaction was monitored by thin layer chromatography (TLC). After completion of the reaction, the reaction mass was cooled to room temperature, filtered through celite and the filtrate was evaporated under reduced pressure. The residue was quenched with ice cold water (50 ml) and extracted with ethyl acetate (EtOAc). The organic phase was dried oversodium sulphate (Na2SO4) and evaporated under reduced pressure to result in 5- bromonaphthalen-1 -amine (2) (15 g, 85%) as an off-white solid.Step 2: 5-bromo-N, N-dimethylnaphthalen-l-amine (3):

[0097] To a stirred solution of 5-bromonaphthalen-l-amine (2) (15.0 g, 67.56 mmol, 1.0 eq.) in dimethyl formamide (DMF) (75 mL), NaH (sodium hydride, 8.1 g, 222.7 mmol, 3.0 eq.) was added and stirred at 0 °C. After 30 minutes, methyl iodide (12.58 ml, 222.70 mmol, 3.0 eq.) was added and the reaction mixture was stirred at room temperature for 16 h to obtain a reaction mass. Progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mass was quenched with ice cold water (50 ml) and was extracted with EtOAc. The organic layer was dried over Na2SO4 and evaporated under reduced pressure to obtain a crude material. The crude material was purified by Combi Flash on 40 g column using 1.5% EtOAc in hexane to obtain 5-bromo-N, N-dimethylnaphthalen- l-amine (3) (12.5 g, 74%) as white solid. LC-MS: (M+l)=252.1Step 2Step 3: Methyl 2-(5-(dimethylamino) naphthalen-l-yl) acetate (5):

[0098] To a stirred solution of 5-bromo-N, N-dimethylnaphthalen-l-amine (3) (12.0 g, 48.0 mmol, 1.0 eq.) in xylene (100.0 mL) was added potassium 3-methoxy- 3-oxopropanoate (4) (11.23 g, 72.0 mmol, 1.5 eq), and DMAP (4- dimethylaminopyridine, 0.585 g, 4.8 mmol, 0.1 eq.). The solution was degassedwith nitrogen gas at room temperature for 10 minutes before and after the addition of Pd(Allyl)2C12 (Allylpalladium chloride, 0.35 g,0.96 mmol, 0.02 eq.), and Xanthphos (4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene, 1.66 g, 2.88 mmol, 0.06 eq.). Then the reaction mixture was stirred at 140 °C for 16 h. Progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mass was filtered through celite bed. The filtrate was diluted with EtOAc and washed with water. The organic layer was washed with saturated sodium bicarbonate solution, brine, dried over sodium sulphate and concentrated under reduced pressure. The obtained crude was purified by Combi Flash using 20% EtOAc in Hexane to afford methyl 2-(5 -(dimethyl amino) naphthal en-l-yl) acetate (5) (6.5 g, 55%) as yellow liquid. LC-MS:(M+l)=244.10Step 3Step 4: 2-(5-(dimethylamino)naphthalen-l-yl)acetic add. Lithium salt (6):

[0099] To a stirred solution of methyl 2-(5-(dimethylamino)naphthalen-l- yl)acetate (5) (6.00 g, 24.69 mmol, 1.0 eq.) in tetrahydrofuran-methanol-water mixture (THF-MeOH-H2O in 3: 1 : 1 ratio, 50.0 mL), aqueous lithium hydroxide (LiOH.H2O, 5.18 g, 123.45 mmol, 5.0 eq.) was added. Then the total reaction mixture was stirred at room temperature for 16 h. Progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mass was concentrated under reduced pressure. The residue was diluted with water and washed with EtOAc. The the aqueous layer was acidified with IN hydrochloric acid (HC1) until pH reached 8 and was then extracted with EtOAc. The obtained organic layer was washed with brine, dried over sodium sulphate and concentrated under reduced pressure to obtain lithium salt of 2-(5-(dimethylamino)naphthalen-l- yl)acetic acid, (6) (6.3 g, crude) as yellow liquid. LC-MS:(M+l)=230.1Step 5: Methyl N-(2-(5-(dimethylamino)naphthalen-l-yl)acetyl)-S-trityl-L- cysteinate (8):

[0100] To a stirred solution of 2-(5-(dimethylamino)naphthalen-l-yl)acetic acid. Lithium salt (6) (2.50 g, 10.91 mmol, 1.0 eq.) in DMF (20.0 mL), HATU (6.23 g, 16.36 mmol, 1.5 eq) was added and was stirred at room temperature. After 10 min, methyl S-trityl-L-cysteinate (7) (3.96 g, 10.91 mmol, 1.0 eq) followed by DIPEA (5.71 mL,32.77mmol, 3.0 eq.) were added and the reaction mixture was stirred at room temperature for 16 h. Progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mass was concentrated under reduced pressure, washed with n-pentane and concentrated under reduced pressure. The crude was purified by combi flash using 30-35% EtOAc in hexane to afford methyl N-(2-(5-(dimethylamino)naphthalen-l-yl)acetyl)-S-trityl-L-cysteinate (8) (5.19 g, 80%) as yellow solid. LC-MS:(M+1) =589.3.Step 6: N-(2-(5-(dimethylamino) naphthalen-l-yl) acetyl)-S-trityl-L-cysteine (9):

[0101] To a stirred solution of methyl N-(2-(5-(dimethylamino) naphthalen-l-yl) acetyl)-S-trityl-L-cysteinate (8) (1.0 g, 1.70 mmol, 1.0 eq.) in THF-MeOH-ELO (in 3: 1 : 1 ratio, 30.0 mL), LiOH.EEO (0.21 g, 5.1 mmol, 3.0 eq.) was added. Then the total reaction mixture was stirred at room temperature for 3 h. Progress of the reaction was monitored by TLC. After completion of the reaction, the reaction masswas concentrated under reduced pressure. The obtained residue was diluted with water and washed with EtOAc. Then the aqueous layer was acidified with IN HC1 pH~8 and was extracted with EtOAc. The organic layer was washed with brine, dried over sodium sulphate, filtered and concentrated under reduced pressure. The crude was purified by Combi Flash using 45-50% EtOAc in hexane to afford N-(2- (5-(dimethylamino)naphthalen-l-yl)acetyl)-S-trityl-L-cysteine (9) (700 mg, 72%) as an off white solid. LC-MS:(M+1) =575.2.Step 7: (2-(5-(dimethylamino) naphthalen-l-yl) acetyl)-L-cysteine (Dimethylamino NaPhAc):

[0102] To a stirred solution of N-(2-(5-(dimethylamino)naphthalen-l-yl)acetyl)-S- trityl-L-cysteine (9) (1.0 g, 1.74 mmol, 1.0 eq.) in DCM (dichloromethane, 15.0 mL) Triisopropyl silane (1.79 mL, 8.7 mmol, 5.0 eq) was added followed by addition of TFA (trifluoroacetic acid, 5.32 mL,8.70 mmol, 40 eq) at 0 °C and was stirred at room temperature for 30 min. Progress of the reaction was monitored by TLC, After completion of the reaction, the reaction mass was concentrated under reduced pressure, washed with n-Pantane concentrated under reduced pressure. The crude compound was purfied by titurating with ethyl acetate and diethyl ether to afford (2-(5 -(dimethyl amino) naphthalen-l-yl) acetyl)-L-cysteine (Formula (I) Dimethylamino NaPhAc, NNDNAC) (1.9 g, 82%) as white solid.

[0103] The obtained compound of Formula (I) was characterized by NMR and mass spectrum.'H NMR (400 MHz, Methanol-d4) 8 8.18 (d, j=8, 1H), 8.03 (d, j= 8 Hz, 1H), 7.60 to 7.76 (m, 4H), 4.61 (dd, J=4.4 Hz, 6.8 Hz, 1H), 4.17 (s, 2H), 3.30 (s, 6H), 2.93 (ddd, 4.4 Hz, 17.2 Hz, 34 Hz, 2H).13C NMR (100 MHz, Methanol-d4) 8 173.0, 172.4, 143.2, 134.4, 134.1, 129.9, 127.7, 127.1, 126.5, 125.4, 121.0, 117.1, 55.5, 46.5, 40.7, 26.2Mass: 332 (M+) 333 (M+l); 331 (M-H).NaPhAc (2 g)Example 2Method of detection of sensitizer

[0104] The compound of Formula (I) (NNDNAC) was used as a probe for detecting the skin-sensitizing compounds.

[0105] The method of detecting a sensitizer was carried out by mixing the compound of Formula (I) (NNDNAC) with a test compound (p-benzoquinone, 2- methyl-4-isothiazolin 3 -one, farnesal, cinnamyl alcohol, lactic acid, p- phenylenediamine, 4-aminophenol) in the presence of 1,8-diazabicyclo [5.4.0]undec-7-ene (base) to obtain a first solution. The first solution was then incubated for about 1 hour to obtain an incubated solution. The incubated solution was subjected to spectral analysis via liquid chromatographic analysis followed by fluorescence measurement by DAD / FLD detectors (diode array detector / fluorescence detector) to measure the depletion of compound of Formula (I). The intensity of the peak of the compound of Formula (I) was correlated to the depletion % of the compound of Formula (I) (probe compound, NNDNAC). Based on the depletion % of the compound of Formula (I) the test compounds were identified as sensitizers. A depletion of 100% of compound of Formula (I) (absence of peak of NNDNAC in chromatogram) pointed that the test compound was a highly reactive sensitizer. Figure 1 (a, b, and c) depict the chromatograms of the incubated solutions comprising the test compounds p-benzoquinone, 2-methyl-4-isothiazolin 3-one, and famesal, respectively, showing 100%, 98% and 3% depletion of the probe compound (NNDNAC) respectively.

[0106] The test compounds were also subjected to existing assays such as Cysteine- direct peptide reactivity assay (Cys-DPRA), NAC-Amino acid derivative reactivity assay (NAC-ADRA) following OECD 442C guidelines, and % depletion of corresponding probe compounds (Cysteine derivative (peptide) and N-Acetyl cysteine) were measured. Other techniques such as Keratinosens and Local lymph node assay (LLNA) were also conducted on the test compounds and the results are summarized in Table 1 below. Table 1ND-Not Determined

[0107] From Table 1, it was observed that p-benzoquinone and 2-methyl 4- isothiazolin-3-one were strong sensitizers with existing methods. The detection and identification of the test compounds using NNDNAC (Formula (I)) as probecompound of the present invention also showed complete depletion of the probe compound (NNDNAC) after 1 h of incubation, which was in accordance with the existing methods. Therefore, it could be understood that the method of the present invention is equivalent to the conventionally employed techniques but incurred much shorter incubation period. Further, Farnesal was observed to be a moderate sensitizer with Cys-DPRA and NAC-ADRA, whereas the method of the present invention showed minimum depletion of the probe compound (NNDNAC) after 1 h of incubation suggesting Farnesal to be a weak sensitizer. This inference was well supported by the KeratinoSens and LLNA data. Cinnamyl alcohol and lactic acid were found to be least reactive or non-reactive in the present method and was in accordance with the existing methods.

[0108] Furthermore, the results of the existing methods Cys-DPRA and NAC-ADRA wherein the incubation time was about 24 hours, concluded that the test compounds such as p-phenylenediamine, and 4-aminophenol to be strong electrophilic sensitizers. However, the results obtained from the method of the present invention (NNDNAC as probe compound) indicated that these compounds were pro-electrophiles and elicited a strong peptide depletion only when the molecules underwent auto oxidation during the prolonged incubation time in the existing methods (NAC-ADRA and Cys-DPRA). Thus, the present method was able to identify pro-electrophiles from electrophiles.

[0109] Table 2 shows the test results for detecting the skin- sensitizing compounds using the method of the present invention as compared to the existing assay NAC- ADRA, at different incubation time period of 1 hour and 24 hours.Table 2

[0110] From Table 2 it could be observed that the existing assay (NAC-ADRA) showed that all the test compounds did not deplete the peptide at 1 hr of incubation time, however at 24 hours of incubation time the test compounds depleted the peptide. Thus, the existing assay was effective only with 24 hours of incubation time. However, the method of the present invention showed varying % depletion of the probe compound based on incubation time for the different test compounds, which facilitated to categorize electrophile and pro-electrophile.

[0111] Figure 2 (a and b) depict chromatograms of incubated solutions of p- benzoquinone under varying incubated time using the method of the present invention and NAC-ADRA respectively.

[0112] % depletion of the probe compound (NNDNAC) was maximum at 1 hr as well as 24 hours of incubation time, for p-benzoquinone and 2-methyl 4- isothiazolin 3-one, confirming these compounds are electrophiles. Thus, the present method utilizing the probe compound (NNDNAC), detected the electrophilic skinsensitizers within the incubation time of 1 hour and confirmed that the present method was time-efficient as compared to the existing assays which required 24 hours of incubation time.

[0113] Further, the compounds p -phenylenediamine and 4-aminophenol showed no depletion of the probe compound (NNDNAC) at 1 hr of incubation whereas said compounds depleted the probe compound at 24 hours of incubation time, which confirmed that these compounds are pro-electrophiles owing to their self-oxidation.

[0114] Thus, the present method is advantageous as it required minimum incubation time and is efficient in distinguishing pro-electrophiles from electrophiles.Example 2 Method of detecting a sensitizer in a concoction

[0115] The compound of Formula (I) (NNDNAC) was used as a probe for detecting the skin-sensitizing compounds from a concoction. The concoction comprises a formulation and a sensitizer or a non-sensitizer. The formulation comprises at least one active material, and at least two additives.

[0116] The below Table 3 shows the constitution of the formulation without sensitizer.Table 3

[0117] In a container, 0.5mg / mL of the formulation was mixed with 4mM of p- benzoquinone to obtain a concoction A. In another container, 0.5mg / mL of the formulation was mixed with 4mM of dimethyl isophthalate to obtain a concoction B. The concoctions were then separately analysed for the presence of sensitizer using the compound of Formula (I) (NNDNAC).

[0118] The method of detecting a sensitizer in a concoction was carried out by mixing the compound of Formula (I) (NNDNAC) with the formulation in the presence of 1,8-diazabicyclo [5.4.0]undec-7-ene (base) to obtain a mixture. The mixture was then incubated for about 2 hours to obtain an incubated mixture. The incubated mixture was subjected to spectral analysis via liquid chromatographic analysis followed by fluorescence measurement by DAD / FLD detectors (diode array detector / fluorescence detector) to measure the depletion of compound of Formula (I). The intensity of the peak of the compound of Formula (I) was correlated to the depletion % of the compound of Formula (I) (probe compound, NNDNAC). Based on the depletion % of the compound of Formula (I) the test compounds were identified as sensitizers. A depletion of 100% of compound of Formula (I) (absence of peak of NNDNAC in chromatogram) pointed that the test compound was a highly reactive sensitizer. Figure 3 (a and b) depict the chromatograms of the incubated mixtures comprising the concoction A and B respectively, showing 100%, and 1.2% depletion of the probe compound (NNDNAC) respectively.

[0119] Similarly, other concoctions were prepared by mixing the formulation with chemicals such as 2,3 butanedione, phenyl acetaldehyde, 2-methyl-2H-Isothiazol- 3 -one, palmitoyl chloride, imidazolidinyl urea, propyl paraben, glycerol, and 6-methyl coumarin. The concoctions prepared were separately analysed using NNDNAC compound.

[0120] The concoctions were also subjected to existing assays such as NAC-Amino acid derivative reactivity assay (NAC-ADRA) following OECD 442C guidelines, and LLNA (in-vivo prediction). In NAC-ADRA method, the percentage depletion of corresponding probe compound (N-Acetyl cysteine) was measured. The results of each technique of analysis are depicted in Table 4.Table 40121] The Table 4 shows that the method of detecting a sensitizer in the concoction using NNDNAC compound of Formula (I) with just 1 to 2 hours of incubation time provided better detection of a sensitizer or a non- sensitizer than the existing techniques. For example, the LLNA method for detection of sensitizer inthe concoction comprising the formulation and Imidazolidinyl urea was considered to be a weak sensitizer.

[0122] Further, the technical advancement of the disclosed method of detecting a sensitizer in the concoction was compared with the conventionally used NAC-ADRA technique with respect to the incubation time as shown in Table 5. The concoction A was mixed with NNDNAC compound of the present disclosure followed by incubation for 2 hours and the depletion of NNDNAC compound was spectrally measured to be 100%. Meanwhile, the concoction A was analysed using NAC-ADRA method where the incubation time was set for 2 hours, but the depletion of corresponding NAC probe compound was found to be very less of about 4.5%. Thus, in terms of shorter incubation time (1-2 hours) and efficient identification of the sensitizer and pre- sensitizer, the method was advantageous wherein the disclosed compound of Formula (I) NNDNAC was employed as a probe.Table 5

[0123] Further, it was also concluded that the disclosed method of detecting a sensitizer in the concoction using the compound of Formula (I) (NNDNAC) of the present disclosure was advantageous in terms of accurate qualitative detection of sensitizer and in terms of the short incubation time required for obtaining the correct results.ADVANTAGES OF THE PRESENT DISCLOSURE

[0124] The present invention provides a compound of Formula (I) and a probe comprising the compound of Formula (I) for rapid analysis of sensitizers incurringshorter incubation time. The present invention also provides a process of preparing the compound of Formula(I).

[0125] The present invention provides a convenient, economical and time efficient method of detecting skin-sensitizing compounds. The probe of the present method is capable of detecting and identifying pro-electrophilic sensitizers and electrophilic sensitizers. Furthermore, the probe of the present method is capable of detecting sensitizers in a concoction.

Claims

I / We Claim:

1. A compound of Formula (I), its solvates, stereoisomers, enantiomers, racemates or salts thereofFormula (I) wherein A is an aryl ring comprising Cio to C20 atoms;Ri and R2 are independently selected from Ci-6 alkyl, or C6-12 aryl; or Ri and R2 combine together to form heterocyclyl ring having 3 to 8 atoms; m is in a range of 0 to 10; and n is in a range of 0 to 10.

2. The compound as claimed in claim 1, wherein A is selected from naphthalene, anthracene, chrysene, phenanthrene, or pyrene; Ri and R2 are independently Ci-6 alkyl; m is in a range of 0 to 3; and n is in a range of 0 to 3.

3. The compound as claimed in any one of the claims 1 and 2, wherein A is naphthalene, Ri and R2 are independently Ci alkyl; m is 1; and n is 0.

4. A probe for detecting a sensitizer comprising the compound of Formula (I) as claimed in any one of the claims 1 to 3.

5. The probe as claimed in claim 4, wherein the sensitizer is an electrophilic or pro-electrophilic skin-sensitizing compound.

6. A method of detecting a sensitizer, the method comprising: a. mixing the compound of Formula (I) as claimed in any one of the claims 1 to 3 or the probe as claimed in claim 4, with the sensitizer in the presence of a base to obtain a first solution; b. incubating the first solution for a time period of 0.5 to 2 hours to obtain an incubated solution;c. spectral analysis of the incubated solution to measure the compound of Formula (I); and d. identifying the sensitizer.

7. The method as claimed in claim 6, wherein the compound of Formula (I) or the probe is in a concentration of at least 5pM, with respect to the total volume of the first solution.

8. The method as claimed in claim 6, wherein the spectral analysis is carried out by chromatographic and fluorescence detection.

9. The method as claimed in claim 6, wherein identifying the sensitizer is by measuring depletion of the compound of Formula (I).

10. The method as claimed in claim 9, wherein depletion of the compound of Formula (I) categorizes the sensitizer.

11. The method as claimed in any one of the claims 6 to 10, wherein the base is selected from 1,8-diazabicyclo [5.4.0]undec-7-ene (DBU), 1,5- diazabicyclo(4.3.0)non-5-ene (DBN), l,4-diazabicyclo[2.2.2]octane (TED), 7-methyl-l,5,7-triazabicyclo[4.4.0] dec-5-ene (MTBD), 1,5,7- triazabicyclo[4.4.0]dec-5-ene (TBD), 4-dimethylamino pyridine, pyridine, piperazine or combinations thereof.

12. The method as claimed in any one of the claims 6 to 11, wherein the sensitizer is an electrophile or a pro-electrophile.

13. A method of detecting a sensitizer in a concoction, the method comprising: a. mixing the compound of Formula (I) as claimed in any one of the claims 1 to 3 or the probe as claimed in claim 4, with the concoction in presence of a base to obtain a mixture; b. incubating the mixture for a time period in a range of 0.5 to 2 hours to obtain an incubated mixture; and c. subjecting the incubated mixture to spectral analysis to detect the presence of the sensitizer.

14. The method as claimed in claim 13, wherein the concoction comprises a formulation comprising at least one active material and at least two additives, and a sensitizer.

15. The method as claimed in claim 13, wherein the compound of Formula (I) or the probe is in a concentration of at least 5pM, with respect to the total volume of the mixture.

16. The method as claimed in claim 13, wherein the sensitizer is in a concentration of at least ImM, with respect to the total volume of the mixture.

17. The method as claimed in claim 13, wherein the formulation is in a concentration range of 0.1 to 2 mg / mL, with respect to the total volume of the mixture.

18. The method as claimed in claim 13, wherein at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or combinations thereof; and at least two additives are selected from a surfactant, a fatty substance, a solvent, a polymer, or a preservative.

19. The method as claimed in claim 13, wherein subjecting the incubated mixture to spectral analysis involves measuring depletion of the compound of Formula (I) or the probe.

20. A composition comprising: a. the compound of Formula (I) as claimed in any one of the claims 1 to 3 or the probe as claimed in claim 4; and b. a formulation comprising at least one active material, and at least two additives; and c. a sensitizer, wherein the compound of Formula (I) or the probe is in a concentration of at least 5pM, with respect to the composition.

21. The composition as claimed in claim 20, wherein at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or combinations thereof; and at least two additives are selected from a surfactant, a fatty substance, a solvent, a polymer, a preservative, or combinations thereof.

22. Use of the compound of Formula (I) as claimed in any one of the claims 1 to 3 or the probe as claimed in claim 4 for detecting a sensitizer.

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