Compounds, probes, and methods thereof

Abstract

The present disclosure provides a compound of Formula (I), wherein A is an aryl ring comprising C 10 to C 20 atoms; R1and R2are independently selected from C 1‑6 alkyl or C 6‑12 aryl; or R1and R2are taken together to form a heterocyclyl ring having 3 to 8 atoms; m is in the range of 0 to 10; and n is in the range of 0 to 10. The present disclosure also provides a probe comprising the compound of Formula (I) for detecting a sensitizer. Furthermore, the present disclosure provides a method of detecting a sensitizer. Formula (I)

Description

Technical Field

[0001] This disclosure generally relates to the analysis and safety assessment of skin-reactive compounds or sensitizers. In particular, this disclosure relates to compounds, probes, and methods for detecting skin sensitizers. Background Technology

[0002] Considering both the nature of the ingredients used and post-market surveillance, skin sensitization is a crucial endpoint in cosmetic risk assessment. Necessary regulatory measures for preventing the induction and triggering of allergies are reflected in several regulatory requirements and common market practices. The coexistence of regulatory needs with recent developments in skin sensitization testing protocols for chemicals, fragrance materials, and cosmetics allows for appropriate hazard and efficacy estimations and risk management without the use of ethically questionable in vivo methods. Due to the complexity of the biological mechanisms associated with skin sensitization, an integrated approach combining different chemical, biological, and computational methods is recommended as an alternative to routine animal testing.

[0003] Chemical methods aim to characterize the potential of sensitizers to induce early molecular initiation events. The presence of electrophilic domains is considered one of the fundamental chemical features for covalently binding biological targets and inducing further hapten processes. Current chemical assays rely on the quantification of unreacted model nucleophiles after incubation with candidate sensitizers.

[0004] Compounds involved in causing allergic contact dermatitis (ACD) are typically electrophilic by nature. Existing techniques or assays utilize the formation of a covalent bond between the electrophile and the nucleophile, due to the interaction of the skin sensitizer with proteins, peptides, and nucleophiles representing those proteins or peptides. However, these techniques result in long incubation times and do not recognize the pro-electrophile.

[0005] Therefore, there is a need to develop reactive compounds that can be used as probes and methods for rapidly identifying potential electrophilic skin sensitizers and distinguishing between electrophilic and pre-electrophilic sensitizers. Invention Overview In one aspect of this disclosure, compounds of formula (I), their solvates, stereoisomers, enantiomers, racemates, or salts thereof are provided: Formula (I) Where A is a subset of C 10 To C 20 The aryl ring of the atom; R1 and R2 are independently selected from C 1-6 Alkyl or C 6-12 Aryl; or R1 and R2 combined to form a heterocyclic ring having 3 to 8 atoms; and m is in the range of 0 to 10; and n is in the range of 0 to 10.

[0007] In another aspect of this disclosure, a probe for detecting sensitizers (which comprises a compound of formula (I) as disclosed herein) is provided.

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

[0009] These and other features, aspects, and advantages of this subject matter will be better understood by referring to the following description and the appended claims. This overview is provided to present the selection of concepts in a simplified form. This overview is not intended to identify key or essential features of the claimed subject matter, nor is it intended to limit the scope of the claimed subject matter.

[0010] Brief description of the attached figures The following figures form part of this specification and are included to further illustrate aspects of this disclosure. A better understanding of this disclosure can be achieved by referring to the accompanying figures in conjunction with the detailed description of the specific embodiments presented herein.

[0011] Figure 1 Chromatograms of compounds of formula (I) according to embodiments of the present disclosure and incubation solutions of (a) p-benzoquinone, (b) 2-methyl-4-isothiazolin-3-one and (c) farnesal are depicted.

[0012] Figure 2 Chromatograms of p-benzoquinone with (a) compound (I) (NNDNAC) and (b) N-(2-(1-naphthyl)acetyl)-L-cysteine ​​(NAC-ADRA) according to embodiments of the present disclosure are depicted at different incubation times of 1 hour and 24 hours.

[0013] Figure 3 A chromatogram is depicted of a compound of formula (I) according to an embodiment of the present disclosure with an incubation solution having (a) a sensitizer of p-benzoquinone and (b) a non-sensitizer of dimethyl isophthalate.

[0014] Description of the present invention Those skilled in the art will recognize that variations and modifications may be made to this disclosure beyond those specifically described. It should be understood that this disclosure includes all such variations and modifications. This disclosure also includes all such steps, features, compositions, and compounds individually or collectively mentioned or indicated in this specification, as well as any and all combinations of any or more such steps or features.

[0015] definition For convenience, certain terms used in the specification and embodiments are described herein before further description of this disclosure. These definitions should be read in accordance with the remainder of this disclosure and as understood by those skilled in the art. The terms used herein have meanings recognized and known to those skilled in the art; however, for convenience and completeness, specific terms and their meanings are set forth below.

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

[0017] The terms “comprise” and “comprising” are used in an inclusive, open sense, meaning that additional elements may be included. They are not intended to be interpreted as “consisting only of”.

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

[0019] Throughout this specification, unless the context otherwise requires, the word “comprise” and its variations such as “comprises” and “comprising” shall be understood to imply inclusion of the stated element or step or group of elements or steps, without excluding any other element or step or group of elements 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 meanings of the various terms used in this description will now be explained.

[0022] According to the implementation scheme described herein, the term "C" 1-6"Alkyl" refers to a straight-chain or branched C1 to C6 alkyl group that can be optionally substituted. Representative examples of alkyl groups include methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl, 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 1-ethyl-2-methylpropyl or different isomers.

[0023] According to the embodiments described herein, the term "aryl" refers to a cyclic or polycyclic (spirocyclic, fused-ring, bridged-ring, non-fused-ring) cyclic group that satisfies Huckel's rule. Representative examples of aryl rings include benzene, naphthalene, phenylene oxide, pyrene, anthracene, phenanthrene, etc.

[0024] According to the embodiments described herein, the terms "heterocyclic" or "heterocyclic" or "heterocyclic group" include "aromatic heterocyclic", "non-aromatic heterocyclic" or polycyclic or bicyclic (spirocyclic, fused-ring, bridged-ring, non-fused-ring) cyclic compounds, wherein the ring may be aromatic or non-aromatic, and wherein the heterocyclic ring contains at least one heteroatom selected from nitrogen, oxygen and sulfur and one or more carbon atoms, totaling three to eight atoms.

[0025] As used herein, the term "solvent" refers to a compound in which its crystal lattice contains one or more solvent molecules. Non-limiting examples of solvates include hydrates when the solvent is water, ammonia when the solvent is ammonia, alcohols when the solvent is an alcohol, and ethers when the solvent is an ether.

[0026] As used herein, the term "salt" refers to a salt of an acid or base of a compound of formula (I). Non-limiting examples of acid salts include inorganic acid salts such as hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, hydroiodic acid, and nitric acid, as well as organic acid salts such as citric acid, fumaric acid, maleic acid, malic acid, mandelic acid, ascorbic acid, oxalic acid, succinic acid, tartaric acid, benzoic acid, acetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid. Non-limiting examples of basic salts include alkali metal salts of sodium hydroxide or potassium hydroxide and alkaline earth metal salts of calcium hydroxide or magnesium hydroxide, and organic bases such as alkylamines, aralkylamines, and heterocyclic amines.

[0027] 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-bonded isomers (i.e., geometric isomers), regioisomers, enantiomers, or diastereomers. Therefore, the chemical structures described herein cover all possible enantiomers and stereoisomers of the shown or identified compounds, including stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and mixtures of enantiomers and stereoisomers. Enantiomers and mixtures of stereoisomers can be separated into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques known to those skilled in the art. Compounds may also exist in several tautomeric forms, including enol forms, ketone forms, and mixtures thereof. Therefore, the chemical structures described herein cover all possible tautomeric forms of the shown or identified compounds. It should also be understood that some isomeric forms, such as diastereomers, enantiomers, and geometric isomers, can be isolated by physical and / or chemical methods and by those skilled in the art. Furthermore, the compound can exist as a mixture of each enantiomer in equal amounts to form a racemic mixture or a racemic mixture.

[0028] According to the embodiments herein, the terms "(one or more) sensitizers" or "(one or more) skin sensitizers" or "(one or more) skin sensitizing compounds" refer to substances that are reactive when applied to the skin and, due to the degree of their reactivity, can cause damage to the skin surface upon contact.

[0029] According to embodiments herein, the term "electrophile" refers to a compound that tends to attract or gain electrons and is thus inherently reactive. For the purposes of this disclosure, the term "electrophile" refers to a "hapten," which is an inherently electrophilic immunogenic compound. These haptens cause allergic reactions upon contact with the skin. For the purposes of this disclosure, electrophiles are electrophilic skin sensitizing compounds and are also referred to as haptens.

[0030] According to the embodiments described herein, the term "pre-electrophile" refers to a compound that is converted to an electrophile upon exposure to certain external conditions. For the purposes of this disclosure, the term "pre-hapten" refers to an inherently non-reactive immunogenic compound that is converted to a hapten upon exposure to certain external conditions. After conversion to a hapten, the pre-hapten causes an allergic reaction on the skin. For the purposes of this disclosure, the pre-electrophile is a pre-electrophilic skin-sensitive compound and is also referred to as a pre-hapten.

[0031] Unless otherwise stated, all percentages, parts, and ratios are based on the total weight of the compositions disclosed herein. Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It should be understood that such range format is used for convenience and brevity only and should be flexibly interpreted to include not only the numerical values ​​explicitly stated as the limits of the range, but also all individual numerical values ​​or subranges contained within that range, as if each numerical value and subrange were explicitly stated. For example, a time period range of about 0.5 to 2 hours should be interpreted to include not only the explicitly stated limits of about 0.5 to about 2 hours, but also subranges such as 0.5 to 0.75 hours, 1.25 to 1.75 hours, etc., and individual quantities (including fractions) within the specified range, such as 0.5h, 1.0h, 1.25h, and 1.75h.

[0032] Unless otherwise defined, 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 pertains. While any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this disclosure, preferred methods and materials are now described. All publications referenced herein are incorporated herein by reference.

[0033] This disclosure is not limited to the specific embodiments described herein, which are intended for illustrative purposes only. Functionally equivalent products and methods are clearly within the scope of this disclosure, as described herein.

[0034] This disclosure provides compounds of formula (I), probes containing compounds of formula (I), and methods for detecting sensitizers using compounds of formula (I). This disclosure provides efficient chemical characterization tools for rapid and high-throughput identification of potential skin sensitizers. The probes and methods for detecting sensitizers disclosed herein are suitable and provide an inexpensive method for pre-screening large numbers of chemical compounds. The method for detecting sensitizers using probes containing compounds of formula (I) is superior to conventional, time-consuming, and expensive in vitro and in vivo assessments of skin-sensitizing compounds. Furthermore, the methods disclosed herein help distinguish between pro-electrophilic and electrophilic sensitizers, which is not possible with existing methods.

[0035] The embodiments described herein provide compounds of formula (I), their solvates, stereoisomers, enantiomers, racemates, or salts thereof. Formula (I) Where A is a subset of C 10 To C 20 The aryl ring of the atom; R1 and R2 are independently selected from C 1-6 Alkyl or C 6-12Aryl; or R1 and R2 combined to form a heterocyclic ring having 3 to 8 atoms; m in the range of 0 to 10; and n in the range of 0 to 10.

[0036] The compounds of formula (I) of the present invention particularly enable the rapid and economical detection of sensitizers. Embodiments herein also provide probes comprising compounds of formula (I).

[0037] The implementation scheme described in this paper also provides a method for detecting sensitizers and classifying proelectropeptides and electrophiles.

[0038] compound According to the present invention, embodiments thereof provide compounds of formula (I), solvates thereof, stereoisomers, enantiomers, racemates thereof, or salts thereof. Formula (I) Where A is a subset of C 10 To C 20 The aryl ring of the atom; R1 and R2 are independently selected from C 1-6 Alkyl or C 6-12 Aryl; or R1 and R2 combined to form a heterocyclic ring having 3 to 8 atoms; m in the range of 0 to 10; and n in the range of 0 to 10.

[0039] According to the embodiments described herein, in particular, it enables the efficient detection of sensitizers using compounds of formula (I).

[0040] According to the specific implementation plan in this paper, A includes C. 10 To C 18 The aryl ring of the atom; R1 and R2 are independently selected from C. 1-6 Alkyl; m is in the range of 0 to 3; and n is in the range of 0 to 3.

[0041] According to the specific implementation scheme described in this paper, A is selected from naphthalene, anthracene, thionyl, phenanthrene, or pyrene; R1 and R2 are independently selected from C. 1-6 Alkyl; m is in the range of 0 to 3; and n is in the range of 0 to 3.

[0042] According to the more specific embodiments described herein, A is naphthalene, R1 and R2 are independently C1 alkyl groups; m is 1; and n is 0.

[0043] According to the embodiments described herein, the compound of formula (I) is used as a nucleophile to react with reactive electrophilic or pre-electrophilic sensitizers, thereby detecting the reactive species. This compound provides specific detection of pre-electrophilic sensitizers within a short incubation period of 0.5 to 2 hours. Furthermore, according to the embodiments described herein, the compound of formula (I) used in the reactive probe overcomes the disadvantages associated with time consumption, difficult detection methods, reagent costs, and high throughput.

[0044] According to other embodiments of this document, the use of a compound of formula (I), its solvates, stereoisomers, enantiomers, racemates, or salts thereof for the detection of skin-sensitizing compounds is provided. According to further embodiments of this document, a compound of formula (I), its solvates, stereoisomers, enantiomers, racemates, or salts thereof are used in a probe for the efficient, rapid, and convenient detection of sensitizers.

[0045] According to other embodiments of this document, the use of compounds of formula (I), their solvates, stereoisomers, enantiomers, racemates, or salts thereof for detecting sensitizers in modulated media is provided. According to further embodiments of this document, compounds of formula (I), their solvates, stereoisomers, enantiomers, racemates, or salts thereof are used in probes for efficient, rapid, and convenient detection of sensitizers.

[0046] probe According to the present invention, embodiments thereof provide a probe for detecting sensitizers comprising a compound of formula (I).

[0047] According to the embodiments described herein, the term "probe" refers to a system used to analyze the properties of a target by reversibly binding to and altering the function of the target. According to another embodiment described herein, the probe is a molecular probe used to detect sensitizers by reacting with them. According to the embodiments described herein, the sensitizer is an electrophile or a pro-electropophile.

[0048] According to other embodiments described herein, the sensitizer is an electrophilic or pre-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. Formula (I) Where A is a subset of C 10 To C 20 The aryl ring of the atom; R1 and R2 are independently selected from C 1-6 Alkyl or C 6-12Aryl; or R1 and R2 combined to form a heterocyclic ring having 3 to 8 atoms; m in the range of 0 to 10; and n in the range of 0 to 10.

[0050] According to the specific implementation scheme described herein, the probe comprises a compound of formula (I), wherein A is a compound containing C. 10 To C 18 The aryl ring of the atom; R1 and R2 are independently selected from C. 1-6 Alkyl; m is in the range of 0 to 3; and n is in the range of 0 to 3.

[0051] According to the specific implementation scheme described herein, the probe comprises a compound of formula (I), wherein A is selected from naphthalene, anthracene, thionylene, phenanthrene, or pyrene; R1 and R2 are independently selected from C 1-6 Alkyl; m is in the range of 0 to 3; and n is in the range of 0 to 3.

[0052] According to other embodiments of this document, the probe comprises a compound of formula (I), wherein A is naphthalene, R1 and R2 are independently C1 alkyl; m is 1; and n is 0.

[0053] According to other embodiments described herein, the probe comprises a compound of formula (I) and other non-reactive components that facilitate the detection of skin-sensitizing compounds. The non-reactive components are inert components that can typically be used in the formulation of the probe.

[0054] According to further embodiments herein, the use of probes comprising compounds of formula (I) for the detection of sensitizers is disclosed. The probes exhibit rapid and efficient detection of electrophilic and pre-electropophilic sensitizers. According to embodiments herein, the probes detect sensitizers using the methods disclosed herein.

[0055] method The implementation scheme described herein includes a method for detecting sensitizers. This method includes detecting sensitizers, particularly electrophilic and pro-electrophophilic sensitizers.

[0056] The embodiments described herein provide a method for detecting sensitizers using a compound of formula (I) as disclosed herein. According to the embodiments described herein, the method for detecting sensitizers includes: (a) mixing a compound of formula (I) with a sensitizer in the presence of a base to obtain a first solution; (b) incubating the first solution for a period of 0.5 to 2 hours to obtain an incubation solution; (c) performing spectroscopic analysis on the incubation solution to measure the compound of formula (I); and (d) identifying the sensitizer.

[0057] The embodiments described herein provide a method for detecting sensitizers using a compound of formula (I) as disclosed herein. According to the embodiments described herein, the method for detecting sensitizers comprises: (a) mixing a compound of formula (I) (where A is naphthalene, R1 and R2 are independently C1 alkyl; m is 1; and n is 0) with a sensitizer in the presence of a base to obtain a first solution; (b) incubating the first solution for a period of 0.5 to 2 hours to obtain an incubation solution; (c) performing spectroscopic analysis on the incubation solution to measure the compound of formula (I); and (d) identifying the sensitizer.

[0058] According to the embodiments described herein, the compound of formula (I) disclosed herein or the probe disclosed herein has a concentration of at least 5 μM relative to the total volume of the first solution. According to specific embodiments described herein, the compound of formula (I) disclosed herein or the probe disclosed herein has a concentration in the range of 5 to 50 μM relative to the total volume of the first solution.

[0059] According to the embodiments described herein, the sensitizer has a concentration of at least 1 mM relative to the total volume of the first solution. According to the specific embodiments described herein, the sensitizer has a concentration in the range of 1 to 10 mM relative to the total volume of the first solution.

[0060] The embodiments described herein provide a method for detecting sensitizers in a modulated article using a compound of formula (I) as disclosed herein. According to the embodiments described herein, the method for detecting sensitizers in a modulated article comprises: (a) mixing a compound of formula (I) with the modulated article in the presence of a base to obtain a mixture; (b) incubating the mixture for a period 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.

[0061] The embodiments described herein provide a method for detecting sensitizers in a modulated mixture using a compound of formula (I) as disclosed herein. According to the embodiments described herein, the method for detecting sensitizers in a modulated mixture comprises: (a) mixing a compound of formula (I) (where A is naphthalene, R1 and R2 are independently C1 alkyl; m is 1; and n is 0) with the modulated mixture in the presence of a base to obtain a mixture; (b) incubating the mixture for a period of 0.5 to 2 hours to obtain an incubation mixture; and (c) performing spectral analysis on the incubation mixture to detect the presence of the sensitizer.

[0062] According to the embodiments described herein, the modifier comprises a formulation and an sensitizer, and the formulation comprises at least one active material and at least two additives.

[0063] According to embodiments herein, the term "modified compound" refers to a mixture of a formulation and an 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 pre-electropophile.

[0064] According to the embodiments herein, the term "method for detecting sensitizers in a modulator" refers to a method for analyzing or examining the presence of sensitizers in a modulator using a compound of formula (I) as disclosed herein or a probe disclosed herein.

[0065] According to the implementation scheme described herein, the incubation mixture is subjected to spectral analysis by measuring the consumption of the compound of formula (I) to identify the presence of sensitizers.

[0066] According to embodiments herein, the compound of formula (I) disclosed herein or the probe disclosed herein is at a concentration of at least 5 μM relative to the total volume of the mixture. According to specific embodiments herein, the compound of formula (I) disclosed herein or the probe disclosed herein is in the concentration range of 5 to 50 μM relative to the total volume of the mixture.

[0067] According to another embodiment of this document, the concentration of the sensitizer is at least 1 mM relative to the total volume of the mixture. According to a specific embodiment of this document, the concentration of a compound of formula (I) disclosed herein or a probe disclosed herein is in the range of 1 to 5 mM relative to the total volume of the mixture.

[0068] According to the embodiments described herein, the formulation has a concentration range of 0.1 to 2 mg / mL relative to the total volume of the mixture. According to a specific embodiment described herein, the formulation has a concentration range of 0.3 to 1 mg / mL relative to the total volume of the mixture. According to a more specific embodiment described herein, the formulation has a concentration of 0.5 mg / mL relative to the total volume of the mixture.

[0069] According to the embodiments described herein, at least one active material is selected from trisodium ethylenediaminedisuccinate, triethanolamine, or a combination thereof; and at least two additives are selected from surfactants, fatty substances, solvents, polymers, preservatives, or a combination thereof. According to a specific embodiment described herein, the at least one active material is a combination of trisodium ethylenediaminedisuccinate and triethanolamine.

[0070] According to another embodiment of this document, the surfactant is selected from C 15-20 Alkyl esters, C 15-20 alkenyl ester, C 15-20 Alkyl-C 2-10 Alkyl esters or combinations thereof; the fatty substance is selected from C 15-20 Alcohol; the solvent is selected from water, C 5-10 Alkane diol, C 15-20Alkanes or combinations thereof; the polymer is selected from C 2-10 Polyolefins, carbomers, or combinations thereof; and the preservative is selected from C 6-10 Aryl alcohols, C 6-10 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 polyisobutylene, acrylic acid and ethyl acetate / cyclohexane copolymers, or combinations thereof; and the preservative is 2-phenoxymethanol.

[0071] According to the embodiments described herein, 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), 1,4-diazabicyclo[2.2.2]octane (TED), 7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (MTBD), 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD), 4-dimethylaminopyridine, pyridine, piperazine, or combinations thereof. According to a specific embodiment described herein, the base is 1,8-diazabicyclo[5.4.0]undec-7-ene. According to another specific embodiment described herein, the base is 1,5-diazabicyclo[4.3.0]non-5-ene.

[0072] According to another embodiment of this invention, the first solution is incubated for a period of 0.5 to 2 hours. According to a specific embodiment of this invention, the first solution is incubated for a period of 0.75 to 1.5 hours. According to a more specific embodiment of this invention, the first solution is incubated for a period of 1 hour.

[0073] According to further embodiments of this paper, identification of the sensitizer involves measuring the consumption of the compound of formula (I). According to other embodiments of this paper, the compound of formula (I) (which is a nucleophile) reacts with the electrophilic sensitizer, resulting in the consumption of the compound of formula (I).

[0074] According to a further embodiment of this invention, the incubation solution is subjected to spectral analysis by chromatography and fluorescence detection. According to a specific embodiment of this invention, spectral analysis is performed by liquid chromatography followed by fluorescence detection. According to a more specific embodiment of this invention, spectral analysis is performed by liquid chromatography followed by fluorescence detection to measure the presence of the compound of formula (I) in the incubation solution and to correlate it with the consumption of the compound of formula (I). The consumption of the compound of formula (I) classifies the sensitizer as an electrophile or a pro-electropophile.

[0075] According to another embodiment of this document, the method for detecting sensitizers as disclosed herein is provided for the pre-screening of cosmetic compositions. The method disclosed herein is used as an analytical evaluation technique for compositions to understand the reactivity of component compounds that cause skin sensitization.

[0076] Composition Embodiments herein include compositions comprising compounds of formula (I) as disclosed herein or probes as disclosed herein. The composition comprises a modulator, wherein the formulation comprises at least one active material, at least two additives, and at least one sensitizer.

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

[0078] According to the embodiments described herein, the at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or a combination thereof; and at least two additives are selected from surfactants, fatty substances, solvents, polymers, preservatives, or a combination thereof.

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

[0080] According to yet another embodiment of this document, the surfactant is selected from C 15-20 Alkyl esters, C 15-20 alkenyl ester, C 15-20 Alkyl-C 2-10 Alkyl esters or combinations thereof; the fatty substance is selected from C 15-20 Alcohol; the solvent is selected from water, C 5-10 Alkane diol, C 15-20 Alkanes or combinations thereof; the polymer is selected from C 2-10 Polyolefins, carbomers, or combinations thereof; and the preservative is selected from C 6-10 Aryl alcohols, C 6-10 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 polyisobutylene, acrylic acid and ethyl acetate / cyclohexane copolymers, or combinations thereof; and the preservative is 2-phenoxymethanol.

[0081] According to the embodiments described herein, at least one sensitizer is selected from electrophilic sensitizers, proelectrophobic sensitizers, or combinations thereof.

[0082] use The embodiments described herein include the use of compounds of formula (I) as disclosed herein for the detection of sensitizers.

[0083] According to the embodiments described herein, the compound of formula (I) can be used to detect sensitizers. This compound can be used to detect and classify sensitizers as electrophiles or pre-electrophophiles. This compound exhibits rapid and accurate detection and identification of both electrophilic and pre-electrophophilic sensitizers. The compound according to the embodiments described herein detects sensitizers using methods as disclosed herein.

[0084] According to another embodiment of this document, the compound of formula (I) can be used to detect the 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 period of 0.5 to 2 hours to obtain an incubation solution; performing spectral analysis on the incubation solution to measure the compound of formula (I); and identifying the presence of the sensitizer.

[0085] According to another embodiment of this document, the compound may more specifically be a compound for detecting and identifying sensitizers, particularly for qualitative identification if the compound is a sensitizer or non-sensitizer, and for qualitative identification if the sensitizer is an electrophile or pre-electropeptide. Preferably, according to embodiments of this document, the compound is a compound for detecting all types of sensitizers, electrophiles, or pre-electropeptides. Thus, in the various embodiments of this document, the compound may be a probe or a composition.

[0086] According to yet another embodiment of this document, the compound can be used to detect and classify sensitizers in a formulation. The formulation comprises a preparation containing at least one active material and at least two additives and a sensitizer. The formulation can be in any suitable form, such as a gel, liquid gel, or liquid. The formulation can also be a cosmetic product applied directly to the skin.

[0087] According to yet another embodiment of this invention, the compound can be used to detect sensitizers at ambient temperature within a short incubation period ranging from 0.5 to 2 hours.

[0088] The embodiments described herein also include the use of probes comprising compounds of formula (I) for the detection of sensitizers. The probes exhibit rapid and efficient detection of electrophilic and pre-electropophilic sensitizers. According to the embodiments described herein, the probes detect sensitizers using methods as disclosed herein.

[0089] According to one embodiment of this article, a probe containing a compound of formula (I) can be used to detect sensitizers in a modulator.

[0090] According to yet another embodiment of this document, a method for detecting sensitizers as disclosed herein is provided for the use of pre-screening compounds used in the manufacture of cosmetic compositions. The method disclosed herein is used as an analytical evaluation technique to understand the reactivity of specific compounds that cause skin sensitization. If the sensitizer is an electrophile, a pro-electropeptide, or a non-sensitizer, the method disclosed herein is used for the qualitative detection and identification of the sensitizer.

[0091] According to yet another embodiment of this document, the method for detecting allergens in a formulation as disclosed herein is provided for use in the pre-screening of cosmetic compositions.

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

[0093] The present disclosure will now be illustrated by the following examples, which are intended to illustrate the work of the disclosure and are not intended to imply any limitation on the scope of the disclosure. Unless otherwise defined, 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 pertains. Although similar or equivalent methods and materials to those described herein may be used in the practice of the disclosed methods and compositions, exemplary methods, apparatus, and materials are described herein. It should be understood that the present disclosure is not limited to the specific methods and experimental conditions described, as such methods and conditions may be applied.

[0094] Example 1 Compound of formula (I) The compound of formula (I) disclosed herein, NN dimethyl N-(2-(1-naphthyl)acetyl)-L-cysteine ​​(NNDNAC) (having A as naphthalene, R1 and R2 independently as C1 alkyl; m as 1; and n as 0) is prepared by the method described herein. Formula (I).

[0095] Preparation of compounds of formula (I) The compound of formula (I) was prepared by the method shown in Scheme 1. Option 1.

[0096] Step 1: 5-Bromonaphthyl-1-amine (2): To a stirred solution of 1-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.) was added at room temperature, followed by ammonium chloride (42.44 g, 793.6 mmol, 10 eq.) to obtain a reaction mixture. The reaction mixture was heated at 75 °C. o The mixture was stirred for 1 hour to obtain the reaction mixture. The reaction progress was monitored by thin-layer chromatography (TLC). After the reaction was complete, the reaction mixture was cooled to room temperature, filtered through diatomaceous earth, and the filtrate was evaporated under reduced pressure. The residue was quenched with ice-cold water (50 ml) and extracted with ethyl acetate (Et OAc). The organic layer was dried over sodium sulfate (Na2SO4) and evaporated under reduced pressure to give 5-bromonaphthyl-1-amine (2) (15 g, 85%) as a grayish-white solid.

[0097] Step 2: 5-Bromo-N,N-dimethylnaphthyl-1-amine (3): To a stirred solution of 5-bromonaphthyl-1-amine (2) (15.0 g, 67.56 mmol, 1.0 equivalent) in dimethylformamide (DMF) (75 mL), NaH (sodium hydride, 8.1 g, 222.7 mmol, 3.0 equivalent) was added and 0 o Stirring was performed. After 30 minutes, iodomethane (12.58 ml, 222.70 mmol, 3.0 equivalent) was added, and the reaction mixture was stirred at room temperature for 16 h to obtain the reactants. The reaction progress was monitored by TLC. After the reaction was complete, the reactants were quenched with ice-cold water (50 ml) and extracted with EtOAc. The organic layer was dried over Na2SO4 and evaporated under reduced pressure to obtain the crude product. The crude product was purified by Combi Flash on a 40 g column using 1.5% EtOAc in hexane to give 5-bromo-N,N-dimethylnaphthyl-1-amine (3) (12.5 g, 74%) as a white solid. LC-MS: (M+1) = 252.1.

[0098] Step 3: Methyl 2-(5-(dimethylamino)naphth-1-yl)acetate (5) Potassium 3-methoxy-3-oxopropionate (4) (11.23 g, 72.0 mmol, 1.5 equivalent) and DMAP (4-dimethylaminopyridine, 0.585 g, 4.8 mmol, 0.1 equivalent) were added to a stirred solution of 5-bromo-N,N-dimethylnaphthyl-1-amine (3) (12.0 g, 48.0 mmol, 1.5 equivalent) in xylene (100.0 mL). The solution was degassed with nitrogen for 10 min at room temperature before and after the addition of Pd(allyl)₂Cl₂ (allyl palladium chloride, 0.35 g, 0.96 mmol, 0.02 equivalent) and Xanthphos (4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, 1.66 g, 2.88 mmol, 0.06 equivalent). The reaction mixture was then stirred at 140 °C for 16 h. The reaction progress was monitored by TLC. After the reaction was complete, the reactants were filtered through a diatomaceous earth bed. The filtrate was diluted with EtOAc and washed with water. The organic layer was washed with saturated sodium bicarbonate solution and brine, dried over sodium sulfate, and concentrated under reduced pressure. The crude product was purified by Combi Flash using 20% ​​EtOAc in hexane to give methyl 2-(5-(dimethylamino)naphthyl-1-yl)acetate (5) (6.5 g, 55%) as a yellow liquid. LC-MS: (M+1) = 244.10.

[0099] Step 4: 2-(5-(dimethylamino)naphth-1-yl)acetic acid.lithium salt (6): To a stirred solution of methyl 2-(5-(dimethylamino)naphthyl-1-yl)acetate (5) (6.00 g, 24.69 mmol, 1.0 equivalent) in a tetrahydrofuran-methanol-water mixture (THF-MeOH-H2O in a 3:1:1 ratio, 50.0 mL), hydrated lithium hydroxide (LiOH·H2O, 5.18 g, 123.45 mmol, 5.0 equivalent) was added. The total reaction mixture was then stirred at room temperature for 16 h. The reaction progress was monitored by TLC. After the reaction was complete, the reactants were concentrated under reduced pressure. The residue was diluted with water and washed with EtOAc. The aqueous layer was acidified with 1 N hydrochloric acid (HCl) until pH reached 8, and then extracted with EtOAc. The resulting organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to give lithium salt of 2-(5-(dimethylamino)naphthyl-1-yl)acetic acid, (6) (6.3 g, crude), as a yellow liquid. LC-MS: (M+1) = 230.1.

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

[0101] Step 6: N-(2-(5-(dimethylamino)naphth-1-yl)acetyl)-S-triphenylmethyl-L-cysteine ​​(9): LiOH·H2O (0.21 g, 5.1 mmol, 3.0 equivalent) was added to a stirred solution of N-(2-(5-(dimethylamino)naphth-1-yl)acetyl)-S-triphenylmethyl-L-cysteine ​​methyl ester (8) (1.0 g, 1.70 mmol, 1.0 equivalent) in THF-MeOH-H2O (3:1:1 ratio, 30.0 mL). The total reaction mixture was then stirred at room temperature for 3 h. The reaction progress was monitored by TLC. After the reaction was complete, the reactants were concentrated under reduced pressure. The resulting residue was diluted with water and washed with EtOAc. The aqueous layer was then acidified with 1N HCl to pH ~8 and extracted with EtOAc. The organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by Combi Flash using 45-50% EtOAc in hexane to give N-(2-(5-(dimethylamino)naphth-1-yl)acetyl)-S-triphenylmethyl-L-cysteine ​​(9) (700 mg, 72%) as a grayish-white solid. LC-MS: (M+1) = 575.2.

[0102] Step 7: (2-(5-(dimethylamino)naphth-1-yl)acetyl)-L-cysteine ​​(dimethylaminoNaPhAc): To a stirred solution of N-(2-(5-(dimethylamino)naphth-1-yl)acetyl)-S-triphenylmethyl-L-cysteine ​​(9) (1.0 g, 1.74 mmol, 1.0 equivalent) in DCM (dichloromethane, 15.0 mL), triisopropylsilane (1.79 mL, 8.7 mmol, 5.0 equivalent) was added, followed by 0 o TFA (trifluoroacetic acid, 5.32 mL, 8.70 mmol, 40 equivalents) was added at C and stirred at room temperature for 30 minutes. The reaction progress was monitored by TLC. After the reaction was complete, the reactants were concentrated under reduced pressure, washed with n-Pantane, and concentrated again under reduced pressure. The crude compound was purified by titrating with ethyl acetate and diethyl ether to give (2-(5-(dimethylamino)naphth-1-yl)acetyl)-L-cysteine ​​(formula (I) dimethylaminoNaPhAc, NNNDNAC) (1.9 g, 82%) as a white solid.

[0103] The obtained compound of formula (I) was characterized by NMR and mass spectrometry.

[0104] 1 ¹H NMR (400 MHz, methanol-d⁴) δ 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).

[0105] 13 C NMR (100 MHz, methanol-d4) δ 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, 26.2.

[0106] Mass spectrometry: 332 (M+) 333 (M+1); 331 (MH).

[0107] Example 2 Methods for detecting allergens The compound of formula (I) (NNDNAC) is used as a probe for detecting skin sensitizing compounds.

[0108] A method for detecting sensitizers involves mixing a compound of formula (I) (NNDNAC) with test compounds (p-benzoquinone, 2-methyl-4-isothiazolin-3-one, farnesaldehyde, 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 is then incubated for approximately 1 hour to obtain an incubation solution. The incubation solution is analyzed by liquid chromatography followed by spectral analysis using a DAD / FLD detector (diode array detector / fluorescence detector) to measure the consumption of the compound of formula (I). The peak intensity of the compound of formula (I) is correlated with the % consumption of the compound of formula (I) (probe compound, NNNDNAC). Based on the % consumption of the compound of formula (I), the test compound is identified as a sensitizer. 100% consumption of the compound of formula (I) (no peak of NNNDNAC in the chromatogram) indicates that the test compound is a highly reactive sensitizer. Figure 1 (a, b, and c) depict chromatograms of incubation solutions containing the test compounds p-benzoquinone, 2-methyl-4-isothiazolin-3-one, and farnesal, respectively, showing 100%, 98%, and 3% consumption of the probe compound (NNDNAC).

[0109] Existing assays were also performed on the test compounds, such as the Cysteine-Direct Peptide Reactivity Assay (Cys-DPRA) and the NAC-Amino Acid Derivative Reactivity Assay (NAC-ADRA) following OECD 442C guidelines, and the percentage consumption of the corresponding probe compounds (cysteine ​​derivatives (peptides) and N-acetylcysteine) was measured. Other techniques, such as keratinosens and local lymph node assay (LLNA), were also performed on the test compounds, and the results are summarized in Table 1 below.

[0110] Table 1 ND - Not determined.

[0111] As can be observed from Table 1, p-benzoquinone and 2-methyl-4-isothiazolin-3-one are strong sensitizers using existing methods. The detection and identification of the test compounds using NNNADAC (Formula (I)) as the probe compound of the present invention also showed complete consumption of the probe compound (NNDNAC) after 1 h of incubation, consistent with existing methods. Therefore, it can be understood that the method of the present invention is equivalent to conventionally used techniques but with a much shorter incubation period. Furthermore, farnesal was observed to be a moderate sensitizer with Cys-DPRA and NAC-ADRA, while the method of the present invention showed minimal consumption of the probe compound (NNDNAC) after 1 h of incubation, indicating that farnesal is a weak sensitizer. Keratinosensing and LLNA data strongly support this inference. Cinnamyl alcohol and lactate were found to be minimally reactive or non-reactive in this method, consistent with existing methods.

[0112] Furthermore, results from existing methods Cys-DPRA and NAC-ADRA (with incubation times of approximately 24 hours) conclude that tested compounds such as p-phenylenediamine and 4-aminophenol are strongly electrophilic sensitizers. However, results obtained from the method of this invention (using NNNDNAC as the probe compound) indicate that these compounds are pro-electrophiles, and only undergo autoxidation during the extended incubation times in existing methods (NAC-ADRA and Cys-DPRA) to induce strong peptide consumption. Therefore, this method enables the identification of pro-electrophiles from electrophiles.

[0113] Table 2 shows the test results of detecting skin sensitizing compounds using the method of the present invention at different incubation periods of 1 hour and 24 hours, compared with existing assays for NAC-ADRA.

[0114] Table 2 .

[0115] As can be observed from Table 2, the existing assay (NAC-ADRA) shows that all test compounds did not consume peptides during the 1-hour incubation period, however, they did consume peptides during the 24-hour incubation period. Therefore, the existing assay is only valid for 24-hour incubation periods. However, the method of the present invention shows different percentages of peptide consumption of probe compounds based on the incubation time of different test compounds, which helps in the classification of electrophiles and pro-electrophosomes.

[0116] Figure 2 (a) and (b) depict chromatograms of benzoquinone incubation solutions using the method of the present invention and NAC-ADRA at different incubation times.

[0117] For p-benzoquinone and 2-methyl-4-isothiazolin-3-one, the consumption percentage of the probe compound (NNDNAC) was highest at 1 hour and 24 hours of incubation, confirming that these compounds are electrophilic. Therefore, this method using the probe compound (NNDNAC) detected electrophilic skin sensitizers within a 1-hour incubation time, demonstrating its time-efficient nature compared to existing assays requiring a 24-hour incubation time.

[0118] Furthermore, the compounds p-phenylenediamine and 4-aminophenol did not show consumption of the probe compound (NNDNAC) after 1 hour of incubation, while the compounds consumed the probe compound after 24 hours of incubation, confirming that these compounds are proelectropophiles due to their auto-oxidation.

[0119] Therefore, this method is advantageous because it requires minimal incubation time and is effective in distinguishing between the pre-electrophile and the electrophile.

[0120] Example 2 Methods for detecting sensitizers in modulators The compound of formula (I) (NNDNAC) is used as a probe to detect skin-sensitizing compounds from a modulator. The modulator comprises a formulation and a sensitizing or non-sensitizing substance. The formulation comprises at least one active material and at least two additives.

[0121] Table 3 below shows the composition of the formulation without sensitizers.

[0122] Table 3 .

[0123] In one container, 0.5 mg / mL of the formulation was mixed with 4 mM p-benzoquinone to obtain formulation A. In another container, 0.5 mg / mL of the formulation was mixed with 4 mM dimethyl isophthalate to obtain formulation B. The presence of sensitizers was then analyzed in each formulation using the compound of formula (I) (NNDNAC).

[0124] A method for detecting sensitizers in a formulation was performed by mixing a compound of formula (I) (NNDNAC) with a preparation in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene (base). The mixture was then incubated for approximately 2 hours to obtain an incubation mixture. The incubation mixture was analyzed by liquid chromatography followed by spectral analysis by fluorescence measurement using a DAD / FLD detector (diode array detector / fluorescence detector) to measure the consumption of the compound of formula (I). The peak intensity of the compound of formula (I) was correlated with the % consumption of the compound of formula (I) (probe compound, NNNDNAC). Based on the % consumption of the compound of formula (I), the test compound was identified as a sensitizer. 100% consumption of the compound of formula (I) (no peak of NNNDNAC in the chromatogram) indicated that the test compound was a highly reactive sensitizer. Figure 3 (a) and (b) depict chromatograms of the incubation mixtures containing modulators A and B, respectively, showing 100% and 1.2% consumption of the probe compound (NNDNAC).

[0125] Similarly, other modifiers were prepared by mixing the formulation with chemicals such as 2,3-butanedione, phenylacetaldehyde, 2-methyl-2-H-isothiazolidin-3-one, palmitoyl chloride, imidazolidinyl urea, propylparaben, glycerol, and 6-methylcoumarin. The prepared modifiers were analyzed separately using NNNDAC compounds.

[0126] Existing assays were also performed on the modulators, such as NAC-amino acid derivative reactivity assay (NAC-ADRA) and LLNA (in vivo prediction) following OECD 442C guidelines. In the NAC-ADRA method, the percentage consumption of the corresponding probe compound (N-acetylcysteine) was measured. The results for each analytical technique are depicted in Table 4.

[0127] Table 4 .

[0128] Table 4 shows that the method for detecting sensitizers in modulators using NNNDNAC compounds of formula (I) with an incubation time of only 1 to 2 hours provides better detection of sensitizers or non-sensitizers than existing techniques. For example, the LLNA method for detecting sensitizers in modulators containing formulations and imidazolidinyl urea is considered a weak sensitizer.

[0129] Furthermore, the technological advancements of the disclosed method for detecting sensitizers in modulators are compared with those of the conventionally used NAC-ADRA technique in terms of incubation time, as shown in Table 5. Modulator A was mixed with the disclosed NNNDNAC compound and then incubated for 2 hours, with spectral measurements showing 100% consumption of the NNNDNAC compound. Simultaneously, modulator A was analyzed using the NAC-ADRA method, where the incubation time was set to 2 hours, but the consumption of the corresponding NAC probe compound was found to be very low, approximately 4.5%. Therefore, this method is advantageous in terms of shorter incubation times (1-2 hours) and efficient identification of sensitizers and pre-sensitizers, wherein the disclosed compound of formula (I), NNNDNAC, is used as a probe.

[0130] Table 5 .

[0131] Furthermore, it is concluded that the disclosed method for detecting sensitizers in modulators using compounds of formula (I) of this disclosure (NNDNAC) is advantageous in terms of accurate qualitative detection of sensitizers and in terms of the short incubation time required to obtain correct results.

[0132] The advantages of this disclosure This invention provides compounds of formula (I) and probes comprising compounds of formula (I) for rapid analysis of sensitizers (induced by short incubation times). This invention also provides a method for preparing compounds of formula (I).

[0133] This invention provides a convenient, economical, and time-efficient method for detecting skin sensitizing compounds. The probe of this method can detect and identify pre-electrophilic and electrophilic sensitizers. Furthermore, the probe of this method can detect sensitizers in modulated compounds.

Claims

1. A compound of formula (I), its solvates, stereoisomers, enantiomers, racemates, or salts thereof. Formula (I) Where A is a subset of C 10 To C 20 The aryl ring of the atom; R1 and R2 are independently selected from C 1-6 Alkyl or C 6-12 Aryl; or R1 and R2 combined to form a heterocyclic ring with 3 to 8 atoms; m is in the range of 0 to 10; and n is in the range of 0 to 10.

2. The compound according to claim 1, wherein A is selected from naphthalene, anthracene, thionylene, phenanthrene, or pyrene; R1 and R2 are independently C. 1-6 Alkyl; m is in the range of 0 to 3; and n is in the range of 0 to 3.

3. The compound according to any one of claims 1 and 2, wherein A is naphthalene, R1 and R2 are independently C1 alkyl; m is 1; and n is 0.

4. A probe for detecting sensitizers, said probe comprising a compound of formula (I) according to any one of claims 1 to 3.

5. The probe according to claim 4, wherein the sensitizer is an electrophilic or pre-electrophilic skin sensitizing compound.

6. A method for detecting sensitizers, the method comprising: a. Mixing the compound of formula (I) according to any one of claims 1 to 3 or the probe according to claim 4 with the sensitizer in the presence of a base to obtain a first solution; b. Incubate the first solution for a period of 0.5 to 2 hours to obtain an incubation solution; c. Perform spectral analysis on the incubation solution to measure the compound of formula (I); and d. Identify the sensitizers.

7. The method according to claim 6, wherein the compound of formula (I) or the probe is at a concentration of at least 5 μM relative to the total volume of the first solution.

8. The method of claim 6, wherein the spectral analysis is performed by chromatography and fluorescence detection.

9. The method according to claim 6, wherein the sensitizer is identified by measuring the consumption of the compound of formula (I).

10. The method according to claim 9, wherein the consumption of the compound of formula (I) classifies the sensitizer.

11. The method according to any one of 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), 1,4-diazabicyclo[2.2.2]octane (TED), 7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (MTBD), 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD), 4-dimethylaminopyridine, pyridine, piperazine, or combinations thereof.

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

13. A method for detecting sensitizers in a modulator, the method comprising: a. Mixing the compound of formula (I) according to any one of claims 1 to 3 or the probe according to claim 4 with the modulator in the presence of a base to obtain a mixture; b. Incubate the mixture for a period of 0.5 to 2 hours to obtain an incubated mixture; and c. Perform spectral analysis on the incubation mixture to detect the presence of the sensitizer.

14. The method of claim 13, wherein the modifier comprises a formulation and an sensitizer, the formulation comprising at least one active material and at least two additives.

15. The method of claim 13, wherein the compound of formula (I) or the probe is at a concentration of at least 5 μM relative to the total volume of the mixture.

16. The method of claim 13, wherein the sensitizer has a concentration of at least 1 mM relative to the total volume of the mixture.

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

18. The method of claim 13, wherein at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or a combination thereof; and at least two additives are selected from surfactants, fatty substances, solvents, polymers, or preservatives.

19. The method of claim 13, wherein applying spectroscopic analysis to the incubation mixture involves measuring the consumption of the compound of formula (I) or the probe.

20. A composition comprising: a. A compound of formula (I) according to any one of claims 1 to 3 or a probe according to claim 4; and b. A formulation comprising at least one active material and at least two additives; and c. The sensitizer, wherein the concentration is at least 5 μM relative to the compound of formula (I) of the composition or the probe.

21. The composition of claim 20, wherein at least one active material is selected from trisodium ethylenediamine disuccinate, triethanolamine, or a combination thereof; and at least two additives are selected from surfactants, fatty substances, solvents, polymers, preservatives, or a combination thereof.

22. Use of the compound of formula (I) according to any one of claims 1 to 3 or the probe according to claim 4 for the detection of sensitizers.

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