Methods of determining susceptibility of olfactory receptor ligands to adaptation

Abstract

The present disclosure generally provides methods for identifying olfactory receptor ligand that are more or less likely to be susceptible to olfactory adaptation, and the cellular assays related to identifying such ligands. In some embodiments, the ligand is an olfactory receptor agonist, and, in other embodiments, the ligand is an olfactory receptor antagonist. In some embodiments, the methods comprise identifying which agonists are more susceptible or less susceptible to olfactory adaptation based on one or more kinetic parameters determined from measuring the intracellular concentration of cAMP over time.

Description

[0001] METHODS OF DETERMINING SUSCEPTIBILITY OF OLFACTORY RECEPTOR LIGANDS TO ADAPTATION

[0002] TECHNICAL FIELD

[0003] The present disclosure generally provides methods for identifying olfactory receptor ligand that are more or less likely to be susceptible to olfactory adaptation, and the cellular assays related to identifying such ligands. In some embodiments, the ligand is an olfactory receptor agonist, and, in other embodiments, the ligand is an olfactory receptor antagonist. In some embodiments, the methods comprise identifying which agonists are more susceptible or less susceptible to olfactory adaptation based on one or more kinetic parameters determined from measuring the intracellular concentration of cAMP over time.

[0004] DESCRIPTION OF RELATED ART

[0005] People commonly experience the physiological phenomenon of olfactory adaptation. Certain fragrances or odors seem to linger, and one continues to smell them as long as these fragrances or odors are present. But other fragrances or odors seem not to linger, even though they remain present. Thus, humans have a tendency to adapt to certain fragrances and odors but not to others. Adaptation thus results in a decreased perceived intensity of volatile and mixtures thereof over prolonged exposure.

[0006] There has been no singular explanation for such adaptation, or the lack of it. Some have suggested that it is of psychological nature. After all, some people tend to experience olfactory adaptation in situations where others do not. But it has also been suggested that there may be a molecular or cellular explanation as well. A variety of different biochemical mechanisms for adaptation have been proposed.

[0007] There may be many benefits to identifying a biochemical explanation, as it would permit the rapid and efficient identification of compounds that may be susceptible to adaptation and therefore as less useful as fragrances in situations where a long-term olfactory stimulation is desired or more useful in situations where a long-term olfactory stimulation is not desired. Additionally, it would also permit the rapid and efficient identification of compounds that may be less susceptible to adaptation and therefore more useful as fragrances in situations where a long-term olfactory stimulation is desired or less useful as fragrances in situations where a longterm olfactory stimulation is not desired. Otherwise, one is left to test for olfactory adaptation through human sensory testing, which can be time-intensive, expensive, and inefficient.

[0008] Therefore, there is a continuing need to understand if there is a biochemical explanation for adaptation and, if so, if there may be ways to apply such understandings to identify compounds that may be more susceptible or less susceptible to olfactory adaptation in a manner that is quick and efficient. SUMMARY

[0009] The present disclosure relates to the discovery that certain parameters calculated from measuring the intracellular cAMP concentration over time following introduction of an olfactory receptor ligand to a cell having an olfactory receptor expressed, for example, on its surface or functionally expressed, correlate reasonably well with the observation of olfactory adaptation in human sensory testing.

[0010] In a first aspect, the disclosure provides a method of identifying a substance that is susceptible to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an agonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the method further comprises: (d) identifying an adaptation-susceptible olfactory receptor agonist that is susceptible to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) deselecting the adaptation-susceptible olfactory receptor agonist as an adaptation-resistant substance.

[0011] In a second aspect, the disclosure provides a method of identifying a substance that is susceptible to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an antagonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the introducing step comprises introducing the olfactory receptor ligand in the presence of an olfactory agonist of the olfactory receptor. In some embodiments, the method further comprises: (d) identifying an adaptation-susceptible olfactory receptor antagonist that is susceptible to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) deselecting the adaptation-susceptible olfactory receptor antagonist as an adaptation-resistant substance.

[0012] In a third aspect, the disclosure provides a method of identifying a substance that is resistant to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an agonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the method further comprises: (d) identifying an adaptation-resistant olfactory receptor agonist that is resistant to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) selecting the adaptation-resistant olfactory receptor agonist as an adaptation-resistant substance.

[0013] In a fourth aspect, the disclosure provides a method of identifying a substance that is resistant to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an antagonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the introducing step comprises introducing the olfactory receptor ligand in the presence of an olfactory agonist of the olfactory receptor. In some embodiments, the method further comprises: (d) identifying an adaptation-resistant olfactory receptor antagonist that is resistant to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) selecting the adaptation-resistant olfactory receptor antagonist as an adaptation-resistant substance.

[0014] In a fifth aspect, the disclosure provides the use of an adaptation-resistant substance identified or selected by the method of the previous aspects to provide a long-lasting fragrance to a fragranced article. In some embodiments, the fragranced article is a fiber, such as a natural or synthetic fiber. In some such embodiments, the fiber is comprised by a textile article, such as an article of clothing, a towel, bedding, upholstery, and the like. In some embodiments, the fragranced article is a personal care product, such as a household cleaning product, a commercial cleaning product, a dish detergent, a laundry detergent, a fabric softener, a scent booster, a shower gel, a shampoo, a hair conditioner, a hair-styling product, a skin care product, a cosmetic product, a deodorant, an antiperspirant, or a self-tanning product. In some embodiments, the fragranced article is a perfuming composition, such as a fine fragrance composition or a fragrance composition for use in various air-care products, such as plug-in diffusers, candles, perfuming reeds, room sprays, and the like. Further aspects, and embodiments thereof, are set forth below in the Detailed Description, the Drawings, the Abstract, and the Claims.

[0015] BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The following drawings are provided for purposes of illustrating various embodiments of the compositions and methods disclosed herein. The drawings are provided for illustrative purposes only and are not intended to describe any preferred compositions or preferred methods, or to serve as a source of any limitations on the scope of the claimed inventions.

[0017] FIG. 1 shows a typical curve showing the increase and decrease of the intracellular cAMP concentration for a period of time following the binding of a ligand to a G-protein-coupled receptor expressed on its surface.

[0018] FIG. 2 shows the calculated self-adaptation index for each odorant at after initial exposure for each odorant.

[0019] FIG. 3 shows the plots of the measured intracellular cAMP concentration as a function of time following introduction of each of four different odorants.

[0020] FIG. 4 shows a plot of the self-adaptation of each of four odorants obtained by human sensory testing plotted against the calculated offset half-life obtained from measuring the intracellular cAMP concentration as a function of time.

[0021] FIG. 5 shows a plot of the self-adaptation of each of four odorants obtained by human sensory testing plotted against the calculated offset slope obtained from measuring the intracellular cAMP concentration as a function of time for OR7A17.

[0022] FIG. 6 shows a plot of the self-adaptation of each of six odorants obtained by an alternative sensory testing plotted against an offset metric obtained from measuring the intracellular cAMP concentration as a function of time for OR7A17.

[0023] FIG. 7 shows a rank-ordered plot of the self-adaptation of each of three odorants obtained by a sensory testing plotted against an offset metric obtained from measuring the intracellular cAMP concentration as a function of time for OR10K1.

[0024] FIG. 8 shows the results of 3 pairwise sensory evaluations and their corresponding kinetic metric obtained from measuring the intracellular cAMP concentration as a function of time for OR10AB1 P, 0R7C1 , and OR10G4.

[0025] FIG. 9 shows the results of a pairwise sensory evaluations and their corresponding kinetic metric obtained from measuring the intracellular cAMP concentration as a function of time for two separate ORs, OR10J5 and OR8H1.

[0026] FIG. 10 shows a rank-ordered plot of the self-adaptation of each of three odorants obtained by a sensory testing plotted against an offset metric obtained from measuring the intracellular cAMP concentration as a function of time for OR5AN1.

[0027] FIG. 11 shows a violin plot of kinetic metrics for two groups of mixtures known to be adapting or non-adapting in sensory testing. On the left, kinetic metrics resulting from individual components of these mixtures dosed against shared odorant receptors (OR10G3, OR10G4, OR10G7, OR2J3, OR5K1 , and OR11 H4) are plotted. On the right, kinetic metrics resulting from pre-mixed mixtures dosed against shared odorant receptors (OR10G3, OR10G4, OR10G7, OR2J3, 0R5K1, and OR11H4) are plotted.

[0028] FIG. 12 shows the kinetic metrics derived from kinetic responses of cells expressing OR11A1 when exposed to 7 distinct antagonists before being washed and then exposed to the cognate agonist geosmin. The offset kinetics on the left are altered with variable effects to the agonist response (right) suggesting that antagonists have distinct regulatory ramifications when exposed then removed from cells expressing odorant receptors. These regulatory effects are relevant to adaptation that cannot be predicted simply from receptor activation.

[0029] FIG. 13 shows kinetic metrics for 6 odorants when individually exposed to cells expressing an odorant receptor for which they are all agonists. For each odorant the metric decreases when exposed to an internalization blocker similar to an odorant that is predicted to be less adapting. Internalization is a desensitization mechanism by which adaptation may occur demonstrating that the kinetic metric is sensitive to perturbations in desensitization and this may underlie the mechanism of its sensory predictive potential.

[0030] DETAILED DESCRIPTION

[0031] The following Detailed Description sets forth various aspects and embodiments provided herein. The description is to be read from the perspective of the person of ordinary skill in the relevant art. Therefore, information that is well known to such ordinarily skilled artisans is not necessarily included.

[0032] Definitions

[0033] The following terms and phrases have the meanings indicated below, unless otherwise provided herein. This disclosure may employ other terms and phrases not expressly defined herein. Such other terms and phrases have the meanings that they would possess within the context of this disclosure to those of ordinary skill in the art. In some instances, a term or phrase may be defined in the singular or plural. In such instances, it is understood that any term in the singular may include its plural counterpart and vice versa, unless expressly indicated to the contrary.

[0034] As used herein, the term “polypeptide” refers to a molecule composed of amino acid monomers linearly linked by amide bonds (also known as peptide bonds). The term “polypeptide” refers to any chain of two or more amino acids and does not refer to a specific length of the chain. Thus, peptides, dipeptides, tripeptides, oligopeptides, “protein,” “amino acid chain,” or any other term used herein to refer to a chain of two or more amino acids, are included within the definition of “polypeptide,” and the term “polypeptide” may be used instead of, or interchangeably with, any of these terms. As used herein, the term “polynucleotide” refers to a compound that includes a polymer of nucleotides. Each nucleotide is composed of a base, specifically a purine or pyrimidine base (namely, cytosine (C), guanine (G), adenine (A), thymine (T) or uracil (U)), a sugar (namely, deoxyribose or ribose), and a phosphate group. In general, the polynucleotide is described by a sequence of bases, whereby the bases represent the primary structure (linear structure) of the polynucleotide. The sequence of bases is typically represented from 5’ to 3’. Herein, the term polynucleotide encompasses deoxyribonucleic acid (DNA), including, for example, complementary DNA (cDNA) and genomic DNA, ribonucleic acid (RNA), in particular, messenger RNA (mRNA), synthetic forms of DNA or RNA, and mixed polymers including two or more of these molecules. The polynucleotide may be linear or circular. In addition, the term polynucleotide includes both sense and antisense strands, as well as single stranded and double stranded forms. Moreover, the polynucleotides described herein can contain naturally occurring or non-naturally occurring nucleotides.

[0035] As used herein, the term “functional fragment” refers to a portion of a polypeptide sequence to which the ligand binds. Polypeptide sequences often contain certain amino acids that do not actively participate in binding, but which may serve other purposes. In some instances, these non-functioning parts of the polypeptide sequence can be removed or partially replaced, while leaving the functional portion of the sequence intact. These modified proteins are said to comprise a functional fragment of the original polypeptide sequence.

[0036] As used herein, the term “adaptation” refers to a reduction in perceived intensity to a compound or a substance after exposure to a compound or a substance for prolonged periods of time such as minutes, hours or days.

[0037] As used herein, the term “ligand” refers to a compound or substance that binds orthosterically or allosterically as an agonist or an antagonist to an olfactory receptor, when the olfactory receptor expressed in a eukaryotic cell in a cell-based assay.

[0038] As used herein, the term “olfactory receptor agonist” refers to a compound or substance that binds as an agonist to an olfactory receptor, when the olfactory receptor expressed in a eukaryotic cell in a cell-based assay, with an EC50 of no more than 600 pM.

[0039] As used herein, the term “olfactory receptor antagonist” refers to a compound or substance that binds as an antagonist to an olfactory receptor, when the olfactory receptor expressed in a eukaryotic cell in a cell-based assay, with an IC50 of no more than 600 pM.

[0040] As used herein, the term “allele” refers to one of several alternative forms of a gene or polynucleotide sequence at a specific chromosomal location (locus).

[0041] As used herein, the term “human OR7A17 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR7A17 alleles, whose sequence is well known in the art. As used herein, the term “human OR2J3 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR2J3 alleles, whose sequence is well known in the art.

[0042] As used herein, the term “human 0R5K1 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR5K1 alleles, whose sequence is well known in the art.

[0043] As used herein, the term “human OR5AN1 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR5AN1 alleles, whose sequence is well known in the art.

[0044] As used herein, the term “human 0R7C1 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR7C1 alleles, whose sequence is well known in the art.

[0045] As used herein, the term “human OR8H1 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR8H1 alleles, whose sequence is well known in the art.

[0046] As used herein, the term “human OR10AB1P olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR10AB1 P alleles, whose sequence is well known in the art.

[0047] As used herein, the term “human OR10G3 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR10G3 alleles, whose sequence is well known in the art.

[0048] As used herein, the term “human OR10G4 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR10G4 alleles, whose sequence is well known in the art.

[0049] As used herein, the term “human OR10G7 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR10G7 alleles, whose sequence is well known in the art.

[0050] As used herein, the term “human OR10K1 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR10K1 alleles, whose sequence is well known in the art.

[0051] As used herein, the term “human OR10J5 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR10J5 alleles, whose sequence is well known in the art.

[0052] As used herein, the term “human 0R11A1 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR11A1 alleles, whose sequence is well known in the art. As used herein, the term “human 0R11 H4 olfactory receptor” refers to the polypeptide sequence that in humans is encoded by the OR11H4 alleles, whose sequence is well known in the art.

[0053] As used herein, the term “Anthamber Premium” refers to the compound having the name ((+-)-1-(2,3,8,8-tetramethyl-1 ,2,3,4,5,6,7,8-octahydro-2-naphthalenyl)ethenone.

[0054] As used herein, the term “Wolfwood” refers to the compound having the name ((+)-(1S,2S,3S,5R)-2,6,6-trimethylspiro[bicyclo[3.1.1]heptane-3,1'-cyclohexane]-2'-en- 4'-one).

[0055] As used herein, “Ambrox” refers to 3a,6,6,9a-tetramethyl-2,4,5,5a,7,8,9,9b-octahydro- 1H-benzo[e][1]benzofuran.

[0056] As used herein, “Polywood Super” refers to (2RS,4aRS,8aSR)-5,5,8a- trimethyldecahydro-2-naphthalenyl acetate.

[0057] As used herein, “Sylvamber” refers to 1-(1 ,2,3,4,5,6,7,8-octahydro-2,3,8,8-tetramethyl- 2-naphtyl)ethan-1-one.

[0058] As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. For example, reference to “a substituent” encompasses a single substituent as well as two or more substituents, and the like.

[0059] As used herein, “for example,” “for instance,” “such as,” or “including” are meant to introduce examples that further clarify more general subject matter. Unless otherwise expressly indicated, such examples are provided only as an aid for understanding embodiments illustrated in the present disclosure and are not meant to be limiting in any fashion. Nor do these phrases indicate any kind of preference for the disclosed embodiment.

[0060] As used herein, “comprise” or “comprises” or “comprising” or “comprised of” refer to groups that are open, meaning that the group can include additional members in addition to those expressly recited. For example, the phrase, “comprises A” means that A must be present, but that other members can be present too. The terms “include,” “have,” and “composed of” and their grammatical variants have the same meaning. In contrast, “consist of” or “consists of” or “consisting of” refer to groups that are closed. For example, the phrase “consists of A” means that A and only A is present.

[0061] As used herein, “optionally” means that the subsequently described event(s) may or may not occur. In some embodiments, the optional event does not occur. In some other embodiments, the optional event does occur one or more times.

[0062] As used herein, “or” is to be given its broadest reasonable interpretation and is not to be limited to an either / or construction. Thus, the phrase “comprising A or B” means that A can be present and not B, or that B is present and not A, or that A and B are both present. Further, if A, for example, defines a class that can have multiple members, e.g., Ai and A2, then one or more members of the class can be present concurrently. Other terms are defined in other portions of this description, even though not included in this subsection.

[0063] Screening Methods

[0064] In certain aspects, the disclosure provides a method of identifying a substance that is susceptible to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an agonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the method further comprises: (d) identifying an adaptation-susceptible olfactory receptor agonist that is susceptible to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) deselecting the adaptation-susceptible olfactory receptor agonist as an adaptation-resistant substance.

[0065] In certain related aspects, the disclosure provides a method of identifying a substance that is susceptible to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an antagonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the introducing step comprises introducing the olfactory receptor ligand in the presence of an olfactory agonist of the olfactory receptor. In some embodiments, the method further comprises: (d) identifying an adaptation-susceptible olfactory receptor antagonist that is susceptible to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) deselecting the adaptation-susceptible olfactory receptor antagonist as an adaptation-resistant substance.

[0066] In some further related aspects, the disclosure provides a method of identifying a substance that is susceptible to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an agonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the method further comprises: (d) identifying an adaptation-resistant olfactory receptor agonist that is resistant to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) selecting the adaptation-resistant olfactory receptor agonist as an adaptation-resistant substance.

[0067] In some further related aspects, the disclosure provides a method of identifying a substance that is susceptible to olfactory adaptation, the method comprising: (a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell, and wherein the olfactory receptor ligand is an antagonist of the plurality of olfactory receptors; (b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and (c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time. In some embodiments, the introducing step comprises introducing the olfactory receptor ligand in the presence of an olfactory agonist of the olfactory receptor. In some embodiments, the method further comprises: (d) identifying an adaptation-resistant olfactory receptor antagonist that is resistant to olfactory adaptation based on the calculated one or more kinetic parameters. In some further embodiments, the method further comprises: (e) selecting the adaptation-resistant olfactory receptor antagonist as an adaptation-resistant substance.

[0068] The foregoing screening methods are not limited to any particular olfactory receptor or its ligands. It is believed that the methods are generalizable to a wide range of olfactory receptors in addition to those for which working Examples are provided. For example, in some embodiments, the plurality of olfactory receptors are a plurality of human olfactory receptors, such as a plurality of human OR2J3, 0R5AN1, OR5K1 , OR7A17, OR7C1 , OR8H1 , OR10AB1 P, OR10G3, OR10G4, OR10G7, OR10J5, OR10K1 , 0R11A1 , OR11H4, or a polypeptide sequence having an amino acid sequence that is at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 97%, or at least 99%, identical thereto.

[0069] Any suitable olfactory receptor ligand can be used in the screening method, so long as the compound or substance is a ligand of at least one of the plurality of olfactory receptors employed in the method. For example, in embodiments where the plurality of olfactory receptors are a plurality of human OR7A17 olfactory receptors, the olfactory receptor ligand is a human OR7A17 olfactory receptor ligand. In general, the particular ligands of various human olfactory receptors are well known in the art. Moreover, such ligands can be identified without undue experimentation by a person of skill in the art using standard cellular screening assays, where the olfactory receptor is expressed in a eukaryotic cell and test compounds or substances are introduced to the assay and an activation or inhibition dose-response curve is generated. The skilled artisan can readily calculate an EC50 from the resulting activation curve or an IC50 for the inhibition curve. In general, compounds or substances having an activation curve with respect to an olfactory receptor with an EC50 of no more than 600 M are classified as agonists of the olfactory receptor in question. In general, compounds or substances having an inhibition curve with respect to an olfactory receptor with an IC50 of no more than 600 pM are classified as antagonists of the olfactory receptor in question.

[0070] Examples of suitable olfactory receptor ligands for use in the methods described herein include, not are not limited to Wolfwood, Anthamber Premium, Ambrox, Polywood Super, and Sylvamber. In general, the plurality of olfactory receptors is expressed on the surface of a plurality of cells. In general, the eukaryotic cells are engineered to overexpress the polypeptides corresponding to the olfactory receptor agonist, a functional fragment thereof, or a polypeptide having at least 70% equivalence to the foregoing. The eukaryotic cells can be engineered to overexpress any of the plurality of olfactory receptors, such as the human OR7A17 olfactory receptor. In some embodiments, the eukaryotic cells are engineered to overexpress one or more of the polypeptides corresponding to different olfactory receptors.

[0071] In some embodiments, the eukaryotic cell further comprises an exogenous nucleic acid sequence that encodes for a G protein. In some further embodiments, the G protein is the a- subunit Golf or any chimera thereof. In some further embodiments, the G protein is gustducin, such as an a-gustducin. In some other embodiments, the G protein is a promiscuous G protein. Non-limiting examples of promiscuous G proteins include Ga16, Ga15, Ga16gust25, Ga15gust25, Ga16gust44, Ga15gust44, and Ga15-i / 3-5, which are described in detail in U.S. Patent No. 10,107,794. In some other embodiments, the G protein is a G(i / o) protein.

[0072] In some embodiments, the eukaryotic cell further comprises a G protein coupled to the olfactory receptors. In some further embodiments, the G-protein is the a-subunit Golf or any chimera thereof. In some further embodiments, the G protein is gustducin, such as an a- gustducin. In some other embodiments, the G protein is a promiscuous G protein. Non-limiting examples of promiscuous G proteins include Ga16, Ga15, Ga16gust25, Ga15gust25, Ga16gust44, Ga15gust44, and Ga15-i / 3-5, which are described in U.S. Patent No. 10,107,794. In some other embodiments, the G protein is a G(i / o) protein.

[0073] Any suitable cell can be used. For example, human HEK-293 and human U2OS cells are well known in the art and are generally available commercially. In some embodiments, the eukaryotic cells are isolated eukaryotic cells. In general, the eukaryotic cells are contained in an assay. In some embodiments, the eukaryotic cells are adhered to a substrate. Various functional assays can be used, including assays that measure of changes in intracellular calcium cation concentration (for example, Fluorometric Imaging Plate Reader-based Ca2+mobilization assay, FLIPR), intracellular cyclic adenosine monophosphate (cAMP) (for example Homogenous Time Resolved Fluorescence, Pherastar), pERK1 / 2 activation (for example, high content imaging, HCI), and receptor internalization (for example, TRANSFLUOR). A change in olfactory receptor activity by a test compound or by an olfactory receptor ligand indicates modulation of the olfactory receptor by the test compound or the agonist.

[0074] In some instances, cells besides human H EK-293 or human U2OS cells can also be used. Non-limiting examples of such cells include the following cell types: 1A2, ARH-77, RWPE-1, WI-38, EJM, NCI-H1155, L-1236, NCI-H526, JM1 , SHP-77, SNU-878, NCI-H2196, C3A, CA46, SNU-466, KS-1 , SNU-738, MOLP-2, HDLM-2, Pfeiffer, HCC-15, Alexander cells, L-540, KMS-12-BM, JK-1, NCI-H1092, SW 1990, NCI-H1184, SU-DHL-1, Hep 3B2.1-7, P3HR- 1 , NCI-H2029, SU-DHL-5, SNU-1, MOLP-8, SUP-M2, MONO-MAC-1 , SNU-1040, KYM-1, HEC-59, HCC1569, OCI-LY3, Hs 819.T, DU4475, CI-1 , S-117, OVCAR-8, SNU-626, HL-60, SUIT-2, T3M-4, RKO, MOR / CPR, DK-MG, GA-10, OCUM-1, HCT-15, HT, MONO-MAC-6, G-402, Toledo, COV362, SU-DHL-8, Daoy, NCI-H1435, LS513, Hs 839.T, Hs 172.T, BT-483, KMS-21 BM, AGS, NCI-H2172, LC-1 / sq-SF, SNU-201 , NUGC-4, SK-HEP-1 , SUP-B15, SNU-5, HT-1197, SUP-T1 , AMO-1 , KU812, AN3 CA, AML-193, VMRC-RCW, HLE, HuH28, Hs 751.T, NCI-H2110, MEG-01 , MV-4-11, Hep G2, KYSE-30, KALS-1, BICR 6, RMUG-S, JHH-6, Ki-JK, IST-MES1 , HCC-95, HPB-ALL, HSC-3, 697, LOU-NH91, KARPAS-299, GI-1 , COLO 792, SK- N-FI, D341 Med, HGC-27, SR-786, COLO-818, MHH-CALL-2, SF126, NCI-H322, A-253, NCI- H1623, MCF7, HCC-44, FU97, OCI-LY-19, Hs 766T, NCI-H522, RL, HCC1428, RPMI 6666, U-937, NCI-H460, SW 1088, NCI-H1792, NCI-H1693, UACC-257, JHUEM-2, HuT 78, UACC- 893, NCI-H929, A-704, OV56, LN-229, OE19, SK-MEL-24, RD-ES, NCI-H211 , KCI-MOH1 , NCI-H1963, Hs 706.T, ChaGo-K-1 , EPLC-272H, OPM-2, KHM-1 B, A549, HuG1-N, NCI-H508, MHH-CALL-3, SNU-1076, A3 / KAW, MEL-HO, TO 175.T, Caki-1 , Hs 936.T, SK-LU-1 , WM- 983B, K-562, EFE-184, SNU-520, NCI-H2291 , HCC-1195, ABC-1 , KE-39, NH-6, HCC2218, CMK, RS4;11 , KYSE-450, OV7, KYSE-510, SK-UT-1, SNU-C1, OE33, P12-ICHIKAWA, DLD-1, COV434, HuNS1 , SNU-899, SW480, COLO-678, LU99, KOPN-8, NCI-H2227, SW1463, Hs 675.T, JHH-4, NCI-H1703, HEC-1-A, BDCM, MIA PaCa-2, PC-3, TE-15, PK-45H, MKN-45, HCC-366, CAL-29, HEC-50B, CPC-N, KMRC-20, SW1116, EOL-1 , COLO 205, EHEB, YD-38, MC116, SK-N-BE(2), BV-173, NCI-H2347, LU65, RT4, U-87 MG, LK-2, KP-N-YN, HEC-251, NCI-H1651 , GP2d, RERF-LC-MS, NB-4, NCI-H2286, SNU-61, T-47D, huH-1, KYSE-180, ST486, SW 1353, M-07e, KASUMI-1 , YH-13, NCI-H28, GAMG, JeKo-1 , GOS-3, SNU-324, PA- TU-8902, MFE-280, SNU-245, NALM-1, RERF-LC-Sq1, BICR 22, ZR-75-1, COR-L23, SW579, COR-L88, KM12, Hs 611.T, OUMS-23, RERF-LC-Ad1 , NCI-H1385, SK-LMS-1 , COLO-320, BL- 70, GRANTA-519, MCAS, Pane 08.13, AM-38, KMS-11, SIG-M5, SNU-407, JHOS-2, OVCAR- 4, Set-2, OV-90, MeWo, HEL, HT-29, MDA-MB-231 , TOV-21G, NCI-H1355, KMS-27, NALM-6, KMS-26, Caov-4, KASUMI-2, UACC-62, U266B1 , Hs 695T, HT55, BICR 31 , TCC-PAN2, KMS- 20, Hs 578T, RI-1, Hs 606.T, NCI-H1341, THP-1, BCP-1 , Hs 737.T, SW1417, MOLT-4, Raji, ESS-1 , MEL-JUSO, SH-10-TC, Hs 683, ME-1 , EB2, PLC / PRF / 5, NCI-H1339, A4 / Fuk, SEM, HEC-265, IST-MES2, KE-97, NCI-H1437, COLO-704, NCI-H1915, TE-5, NCI-H2023, NCI-H82, T1-73, SNU-840, HuT 102, NCI-H1944, KYSE-520, Kasumi-6, 1321N1 , Hs 742.T, IM95, PL45, CL-40, WM1799, KMM-1, SNU-449, JHUEM-1, KARPAS-620, Loucy, SNU-1079, Daudi, HCC-56, HSC-2, COR-L47, PA-TU-89885, OAW28, COR-L311, L-363, Malme-3M, NOMO-1 , Hs 870.T, SU-DHL-10, Hs 229.T, NCI-H810, KYSE-410, RPMI-8402, SNU-175, EBC-1, RVH- 421 , K029AX, PA-TU-8988T, LXF-289, OVSAHO, CAL-12T, Hs 940.T, MM1-S, SUP-HD1 , LNCaP clone FGC, HSC-4, NU-DHL-1 , NCI-H2228, BEN, CAL-78, Sq-1, NCI-H1793, SNU- C2A, M DA- MB- 134- VI, COV318, KE-37, TYK-nu, MOTN-1 , T98G, SW837, EB1 , Becker, PE / CA-PJ34 (clone C12), Hs 616.T, NCI-H446, WM-88, CHP-126, Calu-1, SNU-283, NCI- H1573, SW 1271 , SNU-16, JHOS-4, ACHN, Calu-3, KMRC-1 , SW 1783, TE-11 , TE-9, HuH-6, P31 / FUJ, HT-1376, NCI-H520, 786-0, KNS-60, Caki-2, OVK18, PL-21 , NCI-H2452, JURL- MK1 , TEN, JHH-7, MDA-MB-157, Calu-6, RKN, NUGC-2, ONS-76, J82, OUMS-27, SNU-1196, Hs 739.T, RPMI-7951, NCI-H854, JHH-5, JVM-2, Hey-A8, 5637, KYSE-140, Capan-2, KYSE- 150, HEC-1-B, BICR 16, HEL 92.1.7, MHH-NB-11 , SNU-387, SK-OV-3, SK-MEL-28, IGROV1, ML-1, HLF-a, CHL-1 , YKG1 , A-204, OCI-M1, 8505C, JVM-3, NCI-H647, DB, COLO-800, PK- 59, FaDu, HLF, OVMANA, EFO-27, PF-382, NCI-H747, LS123, SU-DHL-6, SJRH30, PANC-1, NCI-H2342, KM-H2, DND-41 , HH, HuCCTI, F-36P, DMS 454, Hs 274.T, AU565, NCI-H1666, EN, RH-41, NCI-H1373, NCI-H838, SK-MEL-30, MOLM-6, DEL, NCI-H226, NCI-H1648, NCI- H661 , 143B, Mino, C32, KMS-34, NCI-H1694, SK-ES-1 , UACC-812, GDM-1 , NCI-H23, Pane 02.03, CCF-STTG1 , LOX IMVI, SJSA-1, MDST8, PK-1 , NCI-H716, SU-DHL-4, MPP 89, MJ, COLO 829, PE / CA-PJ15, HD-MY-Z, BxPC-3, WM-793, COLO 668, T84, JHOM-1 , PEER, LS411 N, GMS-10, KMBC-2, RMG-I, KELLY, SNU-761, NALM-19, HEC-151, G-361 , OVTOKO, A-498, SW 900, LCLC-103H, FTC-133, QGP-1, Reh, CMK-11-5, NU-DUL-1, BT-20, Hs 600.T, Hs 604.T, KATO III, SNU-410, NCI-H2126, SK-MEL-5, MDA-MB-468, AsPC-1, HUP-T3, KP-N- SI9s, L-428, SNU-1105, HUP-T4, 769-P, LMSU, NCI-H1869, NCO2, MOLM-16, CAL 27, HCC70, NCI-H1930, COV644, Hs 863.T, HCC-2279, D283 Med, Hs 944.T, HCC1599, MDA- MB-415, HCC2157, NCI-H1618, SNU-308, HCC1954, DMS 153, HPAF-II, T24, CJM, VM- CUB1, UM-UC-3, LAMA-84, NCI-H1734, JHH-2, VMRC-RCZ, MFE-319, MDA-MB-453, SNU- 503, TOV-112D, B-CPAP, GSU, HCC-78, NCI-H2171 , CAMA-1, HEC-108, HCC4006, CAL-85- 1 , NCI-H2122, COLO-699, NCI-H196, LUDLU-1 , SW 780, RPMI 8226, LP-1, PC-14, HuTu 80, T.T, SW948, 22Rv1 , HARA, NCI-H596, IPC-298, SCaBER, NCI-H1838, NB-1 , Hs 934.T, Hs 895.T, DMS 114, KYSE-70, KP-3, KP4, DAN-G, NCI-H2009, OC 316, SCC-25, U-138 MG, RCC10RGB, MFE-296, NCI-H1755, RERF-LC-KJ, 8305C, WSU-DLCL2, ES-2, MSTO-211 H, SCC-15, ZR-75-30, PSN1 , SNU-423, NCI-H2106, TE-1 , UT-7, KMS-28BM, NCI-H2081, SK- MM-2, COLO 741 , OC 314, HCC1395, MOLT-13, LN-18, Pane 10.05, PE / CA-PJ41 (clone D2), Hs 746T, CW-2, SKM-1 , NUGC-3, TE-10, NCI-H358, NCI-H69, BFTC-909, HOS, BICR 18, NCI-H1395, OVKATE, Hs 698.T, EFM-19, COLO-783, MHH-CALL-4, ACC-MESO-1, NCI- H1436, KP-N-RT-BM-1, SK-MEL-31, NCI-H1105, CAL-51 , YD-15, NCI-H2085, NCI-H2444, HCC1187, Hs 939. T, CAL-120, SCC-9, TUHR14TKB, KMRC-2, KG-1-C, ECC10, CGTH-W-1, NCI-H841, C2BBe1, SUP-T11, RCH-ACV, CADO-ES1 , JURKAT, 647-V, SK-MEL-2, MDA-MB- 175-VII, MKN74, SNU-C4, LCLC-97TM1 , SCC-4, BHY, IGR-37, KYO-1, Hs 281.T, TT, TUHR4TKB, HT-1080, NCI-H660, TE 441.T, LS1034, KNS-42, Pane 04.03, HCC1419, AZ- 521, SNG-M, NCI-N87, G-292, clone A141 B1, KPL-1, MDA-MB-361, CL-14, NCI-H2170, HuH-7, RD, NCI-H2066, IGR-1 , TE-14, VCaP, BL-41, SNU-620, SK-MES-1 , MEC-2, NCI-H1299, IGR- 39, RT112 / 84, SF-295, DV-90, A2780, BICR 56, NCI-H510, NCI-H2141 , YD-8, NCI-H2405, TF- 1 , MEC-1 , CCK-81 , NCI-H1048, Hs 822.T, NCI-H2052, K052, CAL-54, Hs 840.T, SW620, SK- CO-1 , BT-474, CL-11, KNS-62, NCI-H1650, G-401 , MOLT-16, SNU-398, COLO-680N, EM-2, Hs 294T, CAL-62, KMRC-3, A101 D, KG-1, BT-549, HT115, A-375, SW-1710, WM-115, KLE, JHUEM-3, MKN7, CHP-212, HCC202, BC-3C, NCI-H1568, KMS-18, PE / CA-PJ49, COLO-849, SIMA, OCI-AML3, GSS, EC-GI-10, EFO-21, RCM-1 , DMS 273, KU-19-19, RERF-GC-1B, SH-4, SK-MEL-3, RERF-LC-Ad2, M059K, JHOM-2B, MDA PCa 2b, Hs 852.T, RL95-2, Pane 03.27, SNU-216, Pane 02.13, CFPAC-1 , SK-N-SH, OCI-AML2, LoVo, SBC-5, NCI-H1876, NCI-H441 , SK-N-AS, COR-L24, HCC38, NCI-H1781, DOHH-2, NCI-H1563, U-251 MG, HPAC, JIMT-1 , U- 2 OS, A-673, TC-71 , NCI-H650, NIH:OVCAR-3, CAS-1, JL-1 , SK-MEL-1 , MDA-MB-4355, Ishikawa (Heraklio) 02 ER-, TE 617.T, SU.86.86, RERF-LC-AI, TT2609-C02, LS 180, YAPC, HDQ-P1, KNS-81 , FU-OV-1 , KP-2, DMS 53, SNU-1272, Detroit 562, 42-MG-BA, L3.3, COLO- 679, NCI-H2087, NCI-H2030, GCT, NCI-H889, Caov-3, MDA-MB-436, NCI-H524, MKN1 , KCL- 22, Capan-1, CML-T1, H4, NCI-H727, Hs 343.T, MHH-ES-1 , NMC-G1, HCC-1171 , REC-1 , Hs 618.T, A172, YD-10B, SW48, MUTZ-5, TE-6, JHH-1 , HCT 116, TE-4, IA-LM, MG-63, NCI- H1975, TALL-1 , HCC1806, HMCB, SCLC-21 H, HCC1500, CL-34, Pane 05.04, SW403, TM-31 , HCC1937, JMSU-1 , DMS 79, SNB-19, NCI-H1836, Li-7, HCC827, 639-V, MOLM-13, SK-BR-3, IMR-32, TUHR10TKB, OAW42, SK-N-MC, TGBC11TKB, NCI-H1581, EFM-192A, YMB-1 , HCC2935, ECC12, HCC-33, DU 145, NCI-H146, SNU-1214, SNU-1077, 23132 / 87, HT-144, SNU-182, Hs 888.T, SNU-475, GCIY, Hs 729, JHOC-5, SW 1573, HEC-6, OCI-AMLS, Hs 688(A). T, Hs 821.T, PCM6, RT-112, SK-N-DZ, SNU-478, SNU-119, HCC1143, NCI-H209, 8- MG-BA, COR-L105, COR-L95, SNU-46, COV504, CAL-148, SNU-05, DBTRG-05MG, BHT- 101 , WM-266-4, BFTC-905, KYSE-270, TE-8, SNU-213, and SH-SYSY.

[0075] The methods disclosed herein include a step of measuring the intracellular cAMP concentration in a cell having an olfactory receptor expressed on its surface for a period of time immediately following introduction of the olfactory receptor ligand (agonist or antagonist) for a period of time. FIG. 1 shows a typical curve showing the increase and decrease of the intracellular cAMP concentration for a period of time following the binding of a ligand to a G protein-coupled receptor expressed on its surface. In the methods set forth herein, the measuring generally continues until the concentration of intracellular cAMP reached a reasonable plateau. For example, in some embodiments, the measuring continues for 20 minutes, or 40 minutes, or 60 minutes, or 80 minutes, or 100 minutes, or 120 minutes following introduction of the olfactory receptor ligand to the olfactory receptor. The intracellular cAMP concentration can be measured at any suitable intervals throughout the duration of the measuring, such as at intervals of 10 seconds, 20 seconds, 30 seconds, 45 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, and the like. Measuring the intracellular cAMP concentration can be carried out by any suitable means, which are generally known by those of skill in the art.

[0076] The methods disclosed herein include a step of calculating one or more kinetic parameters from the measured intracellular cAMP concentration over time. Using standard modeling techniques, the skilled artisan can extract certain kinetic parameters from a curve such as that shown in FIG. 1. These kinetic parameters include: the initial rate, the onset slope, the onset half-life (ti / 2), the peak response, the peak time, the plateau, the offset slope, and the offset half-life (ti / 2). In this case, it was discovered that these kinetic parameters calculated for a particular olfactory receptor ligand show a correlation to the observed adaptation experienced by humans to that ligand.

[0077] In some further embodiments, the methods further comprise selecting adaptationresistant ligands or deselecting adaptation-susceptible ligands for use in various applications where olfactory adaptation may be undesirable.

[0078] The olfactory receptor ligand can be any suitable compound or substance that is amenable for use in cellular screening assays. For example, in some embodiments, the olfactory receptor ligand is an organic compound. In some embodiments, the olfactory receptor ligand is a naturally occurring compound. In some other embodiments, the olfactory receptor ligand is a non-naturally occurring compound. In some embodiments, the olfactory receptor ligand is a peptide, such as a polypeptide or an oligopeptide, an amino acid, an amino acid derivative, an amide, a nucleotide, an oligonucleotide. In some embodiments, the olfactory receptor ligand is a plant or food extract, such as a plant or food extract obtained by bioassay- guided fractionation. In some embodiments, the olfactory receptor ligand is a terpene, a diterpene or a triterpene. In some embodiments, the olfactory receptor ligands is a ribonucleotide or derivative thereof. In some embodiments, the olfactory receptor ligand is an organic molecule having a molecular weight of no more than 1000 amu.

[0079] Uses and Methods

[0080] In other aspects, the disclosure provides use of an olfactory-resistant substance identified or selected by the method of the foregoing aspects to provide a long-lasting fragrance to a fragranced article. The disclosure also provides methods that correspond to each of the foregoing uses. Thus, in certain related aspects, the disclosure provides methods of providing a long-lasting fragrance to a fragranced article, the method comprising introducing to an article an adaptation-resistant substance identified or selected by the method of the foregoing aspects. In certain related aspects, the disclosure provides methods of providing a long-lasting fragrance to a fragranced article, the method comprising removing from an article an adaptation-susceptible substance identified or deselected by the method of the foregoing aspects.

[0081] Any suitable fragranced articles are contemplated. In some embodiments, the fragranced article is a fiber, such as a natural or synthetic fiber. In some such embodiments, the fiber is comprised by a textile article, such as an article of clothing, a towel, bedding, upholstery, and the like. In some embodiments, the fragranced article is a personal care product, such as a household cleaning product, an air freshener, a commercial cleaning product, a dish detergent, a laundry detergent, a fabric softener, a scent booster, a shower gel, a shampoo, a hair conditioner, a hair-styling product, a skin care product, a cosmetic product, a deodorant, an antiperspirant, or a self-tanning product. In some embodiments, the fragranced article is a perfuming composition, such as a fine fragrance composition or a fragrance composition for use in various air-care products, such as plug-in diffusers, candles, perfuming reeds, room sprays, and the like.

[0082] Fiber and Textiles

[0083] In certain related aspects, the disclosure provides uses to impart a long-lasting fragrance to a fiber, the method comprising introducing one or more selected or identified substances to the fiber. In general, the fiber is a textile fiber, such as a natural or synthetic fiber comprised by a textile article, such as an article of clothing, a towel, bedding, upholstery, and the like. In some embodiments, the identified or selected substance is introduced to the fiber as part of a perfuming composition included within a personal care composition, such as those described in further detail below.

[0084] Consumer Care Compositions and Related Uses and Products

[0085] In certain related aspects, the disclosure provides uses to impart a long-lasting fragrance to a consumer care composition, the method comprising introducing one or more selected or identified substances to the consumer care composition.

[0086] Further embodiments of the consumer care compositions referenced in connection with the foregoing methods and uses are set forth in more detail below.

[0087] In certain aspects, the disclosure provides a consumer care composition comprising a plurality of microparticles of the first aspect or any embodiments thereof. In some embodiments, the consumer care composition is in the form of a household cleaning product, an air freshener, a commercial cleaning product, a dish detergent, a laundry detergent, a fabric softener, a scent booster, a shower gel, a shampoo, a hair conditioner, a hair-styling product, a skin care product, a cosmetic product, a deodorant, an antiperspirant, or a self-tanning product.

[0088] In some embodiments, the consumer care composition comprises a plurality of microparticles and at least one active ingredient, such as an active ingredient selected from the group consisting of a cosmetic ingredient, skin caring ingredient, perfume ingredient, flavor ingredient, malodor counteracting ingredient, bactericide ingredient, fungicide ingredient, pharmaceutical or agrochemical ingredient, a sanitizing ingredient, an insect repellent or attractant, and mixtures thereof.

[0089] In some embodiments, the consumer care composition is a perfuming composition. In some such embodiments, the consumer care composition comprises a plurality of microparticles, as defined above where the microparticle is a coacervate core-shell microcapsule whose core comprises at least one perfume compound, and, optionally, at least one perfumery adjuvant or liquid perfumery carrier.

[0090] Liquid perfumery carriers are well known in the art. Some non-limiting examples include an emulsifying system, such as a solvent and a surfactant system, or a solvent commonly used in perfumery. Some non-limiting examples of suitable such solvents include dipropyleneglycol, diethyl phthalate, isopropyl myristate, benzyl benzoate, 2-(2-ethoxyethoxy)-1 -ethanol or ethyl citrate, which are the most commonly used. For the compositions which comprise both a perfumery carrier and a perfumery co-ingredient, other suitable perfumery carriers than those previously specified, can be also ethanol, water / ethanol mixtures, limonene or other terpenes, isoparaffins such as those known under the trademark ISOPAR (Exxon Chemical, Houston, Tex., US) or glycol ethers and glycol ether esters such as those known under the trademark DOWANOL (Dow Chemical Company, Midland, Mich., US). By “perfumery co-ingredient” it is meant here a compound, which is used in a perfuming preparation or a composition to impart a hedonic effect and which is not a microcapsule as defined above. In other words such a co- ingredient, to be considered as being a perfuming one, must be recognized by a person skilled in the art as being able to at least impart or modify in a positive or pleasant way the odor of a composition, and not just as having an odor.

[0091] Perfuming co-ingredients are well known in the art. In general terms, these perfuming co-ingredients belong to chemical classes as varied as alcohols, lactones, aldehydes, ketones, esters, ethers, acetates, nitriles, terpenoids, nitrogenous or sulfurous heterocyclic compounds and essential oils, and said perfuming co-ingredients can be of natural or synthetic origin. Many of these co-ingredients are in any case listed in reference texts such as the book by Arctander, PERFUME AND FL VO CHEMICALS (1969) or its more recent versions, or in other works of a similar nature, as well as in the abundant patent literature in the field of perfumery. It is also understood that said co-ingredients may also be compounds known to release in a controlled manner various types of perfuming compounds. Non-limiting examples of co-ingredients include 4-(dodecylthio)-4-(2,6,6-trimethyl-2-cyclohexen-1-yl)-2-butanone, 4-(dodecylthio)-4- (2,6,6-trimethyl-1-cyclohexen-1-yl)-2-butanone, trans-3-(dodecylthio)-1-(2,6,6-trimethyl-3- cyclohexen-1-yl)-1-butanone, 2-(dodecylthio)octan-4-one, 2-phenylethyl oxo(phenyl)acetate, 3,7-dimethylocta-2,6-dien-1-yl oxo(phenyl)acetate, (Z)-hex-3-en-1-yl oxo(phenyl)acetate, 3,7- dimethyl-2,6-octadien-1-yl hexadecanoate, bis(3,7-dimethylocta-2,6-dien-1-yl) succinate, (2-((2- methylundec-1-en-1-yl)oxy)ethyl)-benzene, 1-methoxy-4-(3-methyl-4-phenethoxybut-3-en-1- yl)benzene, (3-methyl-4-phenethoxybut-3-en-1-yl) benzene, 1-(((Z)-hex-3-en-1-yl)oxy)-2- methylundec-1-ene, (2-((2-methylundec-1-en-1-yl)oxy)ethoxy)benzene, 2-methyl-1-(octan-3- yloxy)undec-1-ene, 1-methoxy-4-(1-phenethoxyprop-1-en-2-yl)benzene, 1-methyl-4-(1- phenethoxyprop- 1 -en-2-yl)benzene, 2-(1 -phenethoxyprop- 1 -en-2-yl)naphthalene, (2- phenethoxyvinyl) benzene, 2-(1-((3,7-dimethyloct-6-en-1-yl)oxy)prop-1-en-2-yl)naphthalene, (2- ((2-pentylcyclopentylidene)-methoxy)ethyl)-benzene, 4-allyl-2-methoxy-1-((2-methoxy-2- phenylvinyl)oxy)benzene, (2-((2-heptyl-cyclopentylidene)methoxy)ethyl)benzene, 1-isopropyl-4- methyl-2-((2-pentyl-cyclopentylidene)methoxy)benzene, 2-methoxy-1-((2- pentylcyclopentylidene)methoxy)-4-propylbenzene, 3-methoxy-4-((2-methoxy-2- phenylvinyl)oxy)benzaldehyde, 4-((2-(hexyloxy)-2-phenylvinyl)oxy)-3-methoxybenzaldehyde or a mixture thereof or a mixture thereof.

[0092] The term “perfumery adjuvant” refers to an ingredient capable of imparting additional added benefit such as a color, a particular light resistance, chemical stability, etc. Such compounds are well known in the art.

[0093] The personal care composition can contain any suitable amount of the microparticles of the present disclosure. In some embodiments, the personal care composition comprises from 0.01 % by weight to 30% by weight of microparticles, based on the total weight of the consumer care composition.

[0094] The consumer care composition can be in any suitable physical state, such as a solid (such as a powder), liquid, or a gas. In some embodiments, the consumer care composition is a liquid. In some such embodiments, the liquid consumer care composition has one or more of the following characteristics: a) comprises from 2% by weight to 65% by weight of a surfactant, based on the total weight of the consumer care composition; b) comprises water or a water-miscible hydrophilic organic solvent; c) is in the form of a microparticle slurry; and d) comprises a non-encapsulated perfume.

[0095] In some embodiments, the consumer care composition is in the form of a powder. In some such embodiments, the powder consumer care composition has one or more of the following characteristics: a) comprises from 2% by weight to 65% by weight of a surfactant, based on the total weight of the consumer care composition; b) is in the form of a microcapsule powder; and c) comprises a perfume powder that is different from any perfume comprised by the microparticles.

[0096] In embodiments where the plurality of microparticles comprise encapsulated cores containing perfume compounds, the consumer care composition comprising these microparticles can be of used in various perfumed consumer products, such as products belonging to fine fragrance or “functional” perfumery. Functional perfumery includes personal care products, including hair care products, body cleansing products, skin care products, hygiene products, as well as home care products, including laundry care products, surface care products, and air care products. The term “perfumed consumer product” broadly refers to any consumer product that is expected to deliver, among different benefits, a perfuming effect to the surface to which it is applied, including, but not limited to, skin, hair, textiles, paper, countertops, sinks, toilets, floors, furniture, or other household surfaces, or in the air, for example, in the case of an air freshener, a room deodorizer, a candle, a reed diffuser, and the like.

[0097] Such perfumed consumer products can contain any other ingredients that are commonly used in the industry. Methods of formulating perfume-containing microcapsules are such products is also well known and can be used to develop formulations of such products containing microparticles of the present disclosure.

[0098] Non-limiting examples of suitable perfumed consumer products include a perfume, such as a fine perfume, a cologne, an after-shave lotion, a body splash, a fabric care product, such as a liquid or solid detergent, tablets and unit dose (single or multi-chambers), a fabric softener, a dryer sheet, a fabric refresher, an ironing water, a bleach, a hair care product, such as a shampoo, a hair conditioner, a coloring preparation, or a hair spray, a cosmetic preparation, such as a vanishing cream, a body lotion, or a deodorant or antiperspirant, or a skin-care product, such as a perfumed soap, a shower or bath mousse, a body wash, an oil or gel, bath salts, or a hygiene product, an air care product, such as an air freshener or a “ready to use” powdered air freshener, or a home care product, such all-purpose cleaners, liquid or power or tablet dishwashing products, toilet cleaners, or products for cleaning various surfaces, for example sprays and wipes intended for the treatment or refreshment of textiles or hard surfaces like floors, tiles, and stone, or a hygiene product such as sanitary napkins, diapers, or toilet paper.

[0099] In some embodiments, the consumer care composition comprises: a personal care active base, and a plurality of microparticles according to any of the embodiments set forth herein, wherein the consumer care composition is in the form of a personal care product. Any suitable personal care active base or combination of such materials can be used. Such materials are well known to the skilled artisan and are widely discussed in the relevant patent literature. Non-limiting examples of a consumer care active base include surfactants, oils, hydrophobic solvents, hydrophilic solvents, water, and the like, as well as auxiliary agents, such as bleaching agents, buffering agent, builders, soil release or soil suspension polymers, granulated enzyme particles, corrosion inhibitors, antifoaming, sud suppressing agents, dyes, fillers, and mixtures thereof.

[0100] In some embodiments, the personal care product is a hair-care product, such as a shampoo, a hair conditioner, a coloring preparation, or a hair spray, a cosmetic preparation such as a vanishing cream, a body lotion, or a deodorant or antiperspirant, or a skin care product, such as a perfumed soap, a shower, or a bath mousse, a body wash, an oil or gel, bath salts, or a hygiene product.

[0101] In some embodiments, the consumer care composition comprises: a home care active base, and a plurality of microparticles according to any of the embodiments set forth herein, wherein the consumer care composition is in the form of a home care product.

[0102] Any suitable home care active base or combination of such materials can be used. Such materials are well known to the skilled artisan and are widely discussed in the relevant patent literature. Non-limiting examples of a consumer care active base include surfactants, oils, hydrophobic solvents, hydrophilic solvents, water, and the like, as well as auxiliary agents, such as bleaching agents, buffering agent, builders, soil release or soil suspension polymers, granulated enzyme particles, corrosion inhibitors, antifoaming, sud suppressing agents, dyes, fillers, and mixtures thereof.

[0103] In some embodiments, the home care product is an air care product, such as an air freshener or a “ready to use” powdered air freshener, or a home care product, such all-purpose cleaners, liquid or power or tablet dishwashing products, toilet cleaners, or products for cleaning various surfaces, for example sprays and wipes intended for the treatment or refreshment of textiles or hard surfaces like floors, tiles, and stone, or a hygiene product such as sanitary napkins, diapers, or toilet paper.

[0104] In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition.

[0105] The consumer care composition can have any suitable pH. For example, in some embodiments, the consumer care composition has a pH of less than 7. In some other embodiments, the consumer care product has a pH of at least 7. Fabric Softener

[0106] In some embodiments, the consumer care composition is in the form of a fabric softener composition. In some such embodiments, the consumer care composition comprises a fabric softener active base and a plurality of microparticles according to any of the embodiments set forth above.

[0107] Any suitable fabric softener active base can be used. For example, in some embodiments, the fabric softener active base includes dialkyl quaternary ammonium salts, dialkyl ester quaternary ammonium salts, 1 ,2-dioleoyl-3-trimethylammonium propane, triethanolamine quaternary salts, silicones, and mixtures thereof, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the fabric softener active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises nonencapsulated perfume compounds.

[0108] Liquid Detergent

[0109] In some embodiments, the consumer care composition is in the form of a liquid detergent composition. In some such embodiments, the consumer care composition comprises a liquid detergent active base and a plurality of microparticles according to any of the embodiments set forth above.

[0110] Any suitable liquid detergent active base can be used. For example, in some embodiments, the liquid detergent active base includes anionic surfactant such as alkylbenzenesulfonate (ABS), secondary alkyl sulfonate (SAS), primary alcohol sulfate (PAS), lauryl ether sulfate (LES), methyl ester sulfonate (MES) and nonionic surfactant such as alkyl amines, alkanolamide, fatty alcohol poly(ethylene glycol) ether, fatty alcohol ethoxylate (FAE), ethylene oxide (EO) and propylene oxide (PO) copolymers, amine oxydes, alkyl polyglucosides, alkyl polyglucosamides, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the liquid detergent active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises non-encapsulated perfume compounds. Solid Detergent

[0111] In some embodiments, the consumer care composition is in the form of a solid detergent composition. In some such embodiments, the consumer care composition comprises a solid detergent active base and a plurality of microparticles according to any of the embodiments set forth above.

[0112] Any suitable solid detergent active base can be used. For example, in some embodiments, the solid detergent active base includes anionic surfactant such as alkylbenzenesulfonate (ABS), secondary alkyl sulfonate (SAS), primary alcohol sulfate (PAS), lauryl ether sulfate (LES), methyl ester sulfonate (MES) and nonionic surfactant such as alkyl amines, alkanolamide, fatty alcohol poly(ethylene glycol) ether, fatty alcohol ethoxylate (FAE), ethylene oxide (EO) and propylene oxide (PO) copolymers, amine oxydes, alkyl polyglucosides, alkyl polyglucosamides, as well as various surfactants, hydrophilic organic solvents, water, and fatty acid carboxylates. In some embodiments, the consumer care composition comprises the solid detergent active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises non-encapsulated perfume compounds.

[0113] Shampoo or Shower Gel

[0114] In some embodiments, the consumer care composition is in the form of a shampoo or shower gel composition. In some such embodiments, the consumer care composition comprises a shampoo or shower gel active base and a plurality of microparticles according to any of the embodiments set forth above.

[0115] Any suitable shampoo or shower gel active base can be used. For example, in some embodiments, the shampoo or shower gel active base includes anionic surfactant such as sodium alkylether sulfate, ammonium alkylether sulfates, alkylamphoacetate, cocamidopropyl betaine, cocamide MEA, alkylglucosides and aminoacid based surfactants and mixtures thereof, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the shampoo or shower gel active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises non-encapsulated perfume compounds.

[0116] Rinse-Off Conditioner

[0117] In some embodiments, the consumer care composition is in the form of a rinse-off conditioner composition. In some such embodiments, the consumer care composition comprises a rinse-off conditioner active base and a plurality of microparticles according to any of the embodiments set forth above.

[0118] Any suitable rinse-off conditioner active base can be used. For example, in some embodiments, the rinse-off conditioner active base includes cetyltrimonium chloride, stearyl trimonium chloride, benzalkonium chloride, behentrimonium chloride and mixture thereof, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the rinse-off conditioner active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1 % by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises non-encapsulated perfume compounds.

[0119] Solid Scent Booster

[0120] In some embodiments, the consumer care composition is in the form of a solid scent booster composition. In some such embodiments, the consumer care composition comprises a solid scent booster active base and a plurality of microparticles according to any of the embodiments set forth above.

[0121] Any suitable solid scent booster active base can be used. For example, in some embodiments, the solid scent booster active base includes urea, sodium chloride, sodium sulphate, sodium acetate, zeolite, sodium carbonate, sodium bicarbonate, clay, talc, calcium carbonate, magnesium sulfate, gypsum, calcium sulfate, magnesium oxide, zinc oxide, titanium dioxide, calcium chloride, potassium chloride, magnesium chloride, zinc chloride, saccharides such as sucrose, mono-, di-, and polysaccharides and derivatives such as starch, cellulose, methyl cellulose, ethyl cellulose, propyl cellulose, polyols / sugar alcohols such as sorbitol, maltitol, xylitol, erythritol, and isomalt, PEG, PVP, citric acid or any water soluble solid acid, fatty alcohols or fatty acids and mixtures thereof, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the solid scent booster active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises non-encapsulated perfume compounds.

[0122] Liquid Scent Booster

[0123] In some embodiments, the consumer care composition is in the form of a liquid scent booster composition. In some such embodiments, the consumer care composition comprises a liquid scent booster active base and a plurality of microparticles according to any of the embodiments set forth above.

[0124] Any suitable liquid scent booster active base can be used. For example, in some embodiments, the liquid scent booster active base includes ethoxylated aliphatic alcohols, POE / PPG (polyoxyethylene and polyoxypropylene) ethers, mono and polyglyceryl esters, sucrose ester compounds, polyoxyethylene hydroxylesters, alkyl polyglucosides, amine oxides, alcohols, salts and esters of carboxylic acids, salts and esters of hydroxyl carboxylic acids, fatty acids, fatty acid salts, glycerol fatty acids, and mixtures thereof, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the liquid scent booster active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises nonencapsulated perfume compounds.

[0125] Hair Colorant

[0126] In some embodiments, the consumer care composition is in the form of a hair colorant composition. In some such embodiments, the consumer care composition comprises a hair colorant active base and a plurality of microparticles according to any of the embodiments set forth above.

[0127] Any suitable hair colorant active base can be used. For example, in some embodiments, the hair colorant active base includes oxidizing agents, an alkakine agent, dye precursors, coupling agents, as well as various surfactants, hydrophilic organic solvents, and water. In some embodiments, the consumer care composition comprises the hair colorant active base in an amount ranging from 85% by weight and 99.95% by weight, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 15 % by weight, or from 0.2% by weight to 5% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the consumer care composition comprises non-encapsulated perfume compounds.

[0128] Perfuming Composition

[0129] In some embodiments, the consumer care composition is in the form of a perfuming composition. In some such embodiments, the consumer care composition comprises perfume compounds, ethanol, and a plurality of microparticles according to any of the embodiments set forth above. In some embodiments, the consumer care composition comprises the plurality of microparticles in an amount ranging from 0.1% by weight to 30 % by weight, or from 0.2% by weight to 20% by weight, of the plurality of microparticles, based on the total weight of the consumer care composition. In some embodiments, the perfume compounds are present in an amount ranging from 0% by weight to 40% by weight, or from 3% by weight to 40% by weight, based on the total weight of the consumer care composition. In some embodiments, the ethanol is present in an amount ranging from 20% by weight to 90% by weight, or from 40% by weight to 90% by weight, based on the total weight of the consumer care composition.

[0130] EXAMPLES

[0131] To further illustrate this invention, the following examples are included. The examples should not, of course, be construed as specifically limiting the invention. Variations of these examples within the scope of the claims are within the purview of one skilled in the art and are considered to fall within the scope of the invention as described and claimed herein. The reader will recognize that the skilled artisan, armed with the present disclosure, and skill in the art is able to prepare and use the invention without exhaustive examples.

[0132] Example 1 - Sensory Testing Using Air Diffusion Booths

[0133] To validate findings from the cell-based assays, sensory testing was performed using human subjects. A device was used to diffuse either Wolfwood, Ambrox, Polywood Super, or Sylvamber in a sensory evaluation booth. Each panelist was asked to evaluate the intensity of the odorant at relevant time intervals. FIG. 2 shows the adaptation index for the four odorants based on human sensory evaluations.

[0134] Example 2 - Measurement of Intracellular cAMP and Calculation of Kinetic Parameters

[0135] Eukaryotic cells were transfected with a human OR7A17 olfactory receptor and Golf and were treated with four prospective OR7A17 agonists. The four agonists used were Wolfwood, Ambrox, Polywood Super, and Sylvamber. The intracellular cAMP concentration was measured for a period of 4000 seconds. FIG. 3 shows the plots of the measured intracellular cAMP concentration following introduction of each of the four odorants.

[0136] The kinetic parameters were calculated for each of the four concentration plots. These parameters included but were not limited to offset half-life, offset slope, and peak time. FIG. 4 shows a plot of the self-adaptation of each of the four odorants obtained by human sensory testing using booth methodology from Example 1 plotted against the calculated offset half-life. A linear correlation analysis provided an R2of 0.55. FIG. 5 shows a plot of the self-adaptation of each of the four odorants obtained by human sensory testing plotted against the calculated offset slope. A linear correlation analysis provided an R2of 0.55.

[0137] Example 3 - Comparing Kinetic Parameters of Individual OR Agonists to Olfactometry-Based Human Sensory Studies

[0138] A second human sensory assay assessing adaptation, distinct from that used in Example 1 , was developed. This assay measures the decrease in perceived intensity (A) of a stimulus after prolonged minutes-long exposure to an adaptant using olfactometry using >20 panelists per experiment. The method includes QC metrics such as breathing freguency normalization, individual iso-intensity normalization, blinding, and order-based bias assessment. A larger delta measurement collected from this method indicates that the odorant is more highly adapted to. In Example 3, the adaptant and stimulus were the same compound, which is indicative of selfadaptation. T o demonstrate the applicability of kinetics prediction of self-adaptation to the broader family of odorant receptors, kinetic parameters derived using the method described in Example 2 were compared to olfactometer-based adaptation measurements for agonists of OR7A17, OR10K1, OR10AB1P, OR7C1 , OR10G4, OR10J5, OR8H1, and OR5AN1. In each of these examples, there was a direct correlation of offset metrics to sensory adaptation (FIG. 6 - FIG. 10). These results indicate that this in vitro methodology can serve as a predictor of in vivo adaptation.

[0139] Example 4 - Comparing Kinetic Parameters of Mixtures of OR Agonists to Olfactometry-Based Human Sensory Studies

[0140] In addition to the prediction of adaptation for single fragrance ingredients as described in the Examples 1-3, it would be of use to predict adaptation of complex mixtures of fragrance ingredients. We sought to identify two mixtures of 5 ingredients each that have differentiated adaptation in human sensory testing to validate whether our in vitro method could indeed predict their sensory adaptation. Using the olfactometry-based human sensory method described in Example 3, two mixtures of a similar scent camp were identified where one mixture was ~2 fold more adapting than the other. Using historical data, 5 Ors with demonstrated agonism by at least 1 component of each mixture were selected to be tested in vitro. Kinetic parameters for each of these Ors were collected in response to the individual components of the mixtures as well as the holistic mixture. The offset metrics for individual OR-agonist pairs were directly correlated with the sensory adaptation of the mixture (FIG. 11 , left). Additionally, the offset metrics for the OR- mixture combination were directly correlated with the sensory adaptation of the mixture (FIG. 11 , right). These results indicate that kinetic parameters of the individual components of a mixture or the holistic mixture itself can be predictive of the mixture’s sensory adaptation in vivo.

[0141] Example 5 - Kinetic Parameters of OR Antagonists Alter OR

[0142] Compounds that bind receptors but do not agonize the receptor, including antagonists, have the potential to affect the regulatory fate of the bound receptor including desensitization. Kinetic parameters of individual OR agonists, as shown in Examples 2-3, can predict regulatory fate of the OR i.e., self-adaptation. To understand if kinetic parameters can also describe regulatory outcomes of non-agonists, a modified version of the protocol in Example 2 was run in which cells expressing OR11 A1 were pre-treated one of seven compounds for 30 minutes prior to washing and exposure to a OR11A1 agonist. In previous experiments, the modulators were determined to have similar ICso and % inhibition values. In FIG. 12, it is shown that the offset metrics (left) and extent of agonism (right) by the OR11A1 agonist were altered by preincubation with the modulator. Furthermore, the alteration of offset metrics does not correlate to the respective modulation of the agonism suggesting that the modulators have distinct, varied, and lasting effects on subsequent OR activity. These results combined with the predictive potential demonstrated in Examples 1-4 suggest that measuring kinetic parameters in modulator-agonist mixtures could provide prediction of adaptation for mixtures in vivo.

[0143] Example 6 - Pharmacological Blockers of Desensitization Are Predicted by Kinetics Parameters

[0144] Adaptation is a by-product of various biological pathways including desensitization at the periphery where receptors sense small molecules in the olfactory epithelium. A known mechanism of membrane receptor desensitization is receptor internalization. To demonstrate that kinetic parameters are indicative of one such mechanism of desensitization at the molecular level, cells expressing OR7A17 were exposed to small molecules known to block internalization of receptors before being exposed to a variety of OR7A17 agonists (FIG. 13). Cells with blocked internalization universally had lower offset metrics, correlating with a less desensitized system. These results provide mechanistic validation that kinetic parameters can be used to predict the regulatory response to OR binders and provides another example of the the same directionality of this correlation (higher offset metric leads to higher adaptation / desensitization) .

Claims

CLAIMS1. A method of identifying a substance that is susceptible to olfactory adaptation, the method comprising:(a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell;(b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and(c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time.

2. The method of claim 1, wherein the olfactory receptor ligand is an olfactory receptor agonist.

3. The method of claim 1, wherein the olfactory receptor ligand is an olfactory receptor antagonist, and the introducing comprises introducing the olfactory receptor ligand to the plurality of olfactory receptors in the presence of an olfactory receptor agonist.

4. The method of any one of claims 1 to 3, further comprising: (d) identifying an adaptation- susceptible olfactory receptor ligand that is susceptible to olfactory adaptation based on at least one of the calculated one or more kinetic parameters.

5. The method of claim 4, further comprising: (e) deselecting the adaptation-susceptible olfactory receptor ligand as an adaptation-resistant substance.

6. A method of identifying a substance that is resistant to olfactory adaptation, the method comprising:(a) introducing an olfactory receptor ligand to a plurality of olfactory receptors, wherein each of the plurality of olfactory receptors is expressed on a surface of a cell;(b) measuring an intracellular cyclic adenosine monophosphate (cAMP) concentration in one or more cells expressing at least one of the plurality of olfactory receptors over a period of time immediately following introducing the olfactory receptor ligand; and(c) calculating one or more kinetic parameters from the measured intracellular cAMP concentration over the period of time.

7. The method of claim 6, wherein the olfactory receptor ligand is an olfactory receptor agonist.

8. The method of claim 6, wherein the olfactory receptor ligand is an olfactory receptor antagonist, and the introducing comprises introducing the olfactory receptor ligand to the plurality of olfactory receptors in the presence of an olfactory receptor agonist.

9. The method of any one of claims 6 to 8, further comprising: (d) identifying an adaptationresistant olfactory receptor ligand that is resistant to olfactory adaptation based on at least one of the calculated one or more kinetic parameters.

10. The method of claim 9, further comprising: (e) selecting the adaptation-resistant olfactory receptor ligand as an adaptation-resistant substance.

11. The method of any one of claims 1 to 10, wherein the olfactory receptor is selected from the group consisting of: the human OR2J3, OR5AN1 , OR5K1 , OR7A17, OR7C1 , OR8H1 , OR10AB1 P, OR10G3, OR10G4, OR10G7, OR10J5, OR10K1 , OR11A1, OR11H4, and any polypeptide sequence having an amino acid sequence that is at least 70% identical thereto.

12. Use of an adaptation-resistant substance identified or selected by the method of claim 4, 5, 9, or 10 to provide a long-lasting fragrance to a fragranced article.

13. The use of claim 12, wherein the fragranced article is a fiber, such as a synthetic or natural fiber.

14. The use of claim 12, wherein the fragranced article is a consumer care product, such as a household cleaning product, an air freshener, a commercial cleaning product, a dish detergent, a laundry detergent, a fabric softener, a scent booster, a shower gel, a shampoo, a hair conditioner, a hair-styling product, a skin care product, a cosmetic product, a deodorant, an antiperspirant, or a self-tanning product.

15. The use of claim 12, wherein the fragranced article is a perfuming composition, such as a fine fragrance composition.

16. A method of providing a long-lasting fragrance to a fragranced article, the method comprising introducing an adaptation-resistant substance identified or selected by the method of claim 4, 5, 9, or 10 to the fragranced article.

17. A method of providing a long-lasting fragrance to a fragranced article, the method comprising removing an adaptation-susceptible substance identified or selected by the method of claim 2, 3, 7, or 8 to the fragranced article.

18. The method of claim 16 or 17, wherein the fragranced article is a fiber, such as a synthetic or natural fiber.

19. The method of claim 16 or 17, wherein the fragranced article is a consumer care product, such as a household cleaning product, a commercial cleaning product, a dish detergent, a laundry detergent, a fabric softener, a scent booster, a shower gel, a shampoo, a hair conditioner, a hair-styling product, a skin care product, a cosmetic product, a deodorant, an antiperspirant, or a self-tanning product.

20. The method of claim 16 or 17, wherein the fragranced article is a perfuming composition, such as a fine fragrance composition.

21. A perfuming composition for providing long-lasting fragrance, which comprises an adaptation-resistant substance identified or selected by the method of claim 2, 3, 5, or 6.

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