MACROCYCLIC MUSK LACTONES AND USES THEREOF
`
`FIELD OF THE INVENTION
`
`The field of the invention relates to lactone compounds(e.g., macrocyclic lactone
`
`compoundsthat may be flavor- or fragrance-bearing) and their uses (e.g., in fragrance
`
`materials).
`
`BACKGROUND
`
`Musklactones are generally scarce and expensive. Currently, only a limited number of
`
`10
`
`macrocyclic musk lactones are available and only with limited differentiated muskynotes.
`
`Mostof the existing musk lactones are chemically synthesized, and few are natural.
`
`SUMMARY
`
`Many macrolactones, particularly C14-C16 lactones are knownin the art and many have
`
`15
`
`been described as having to a greater or lesser extent a musk-like odor. However, these odors
`
`differ considerably in quality as well as muskintensity and only a few have so far found practical
`
`use in perfumery. Hexadecen-7-olide, also known as Ambrettolide, is a well-known fragrance
`
`material with a musky-floral odor (S. Arctander Perfume and Flavour Chemicals, monograph
`
`105), is probably one of the most extensively used macrocyclic musk materials and is marked
`
`20
`
`under many tradenames, while its isomers with the double bondin the 5 or 6 position only have
`
`a faint musky odor. Hexadecanolide (S. Arctander, monograph 923) is again a well-known musk
`
`fragrance material. There is a need in perfumery for novel musk fragrance materials with
`
`comparable or better odor properties than the ones whichare presently used.
`
`The present disclosure, in some aspects, provide lactones, such as macrocyclic lactones
`
`25
`
`for use in consumer products. As described hercin, the lactones are produced using
`
`biosynthetic methods from fatty acids.
`
`In some embodiments, the lactones have differentiated
`
`musky notes, can be used in consumerproducts comprising fragranced compositions, for
`
`boosting a second fragrance or overall fragrance performance, have mood enhancing and/or
`
`anti-inflammatory effects, and have increased substantivity (e.g., fiber substantivity) and
`
`30
`
`biodegradability.
`
`Accordingly, some aspects of the present disclosure provide fragrance compositions
`
`comprising a lactone comprising one or more compoundsofthe formula:
`
`

`

`wherein:
`
`R is methyl, ethyl, or n-propyl;
`
`each --- is independently a single bond, F double bond, Z double bond, or triple bond,
`
`5
`
`as valency permits; and
`
`k is an integer between 6 and 30,inclusive.
`
`In some embodiments, the lactone comprises one or more compoundsof the formula:
`
`O
`
`ao
`(JoAR,
`
`wherein:
`
`10
`
`R is methyl, ethyl, or n-propyl;
`
`each --- is independently a single bond, £ double bond, Z double bond,ortriple bond,
`
`as valency permits; and
`
`nis an integer between 6 and 20,inclusive.
`
`Otheraspects of the present disclosure provide consumer products comprising a
`
`15
`
`fragrance composition comprising a lactone comprising one or more compoundsof the
`
`formula:
`
`wherein:
`
`R is methyl, ethyl, or n-propyl;
`
`20
`
`each --- is independently a single bond, F double bond, Z double bond, or triple bond,
`
`as valency permits; and
`
`k is an integer between 6 and 30, inclusive.
`
`

`

`In some embodiments, the fragrance composition comprises a lactone comprising one or more
`
`compoundsof the formula:
`
`oO
`
`no
`(< >
`dn
`
`R ,
`
`wherein:
`
`R is methyl, ethyl, or n-propyl;
`
`5
`
`each --- is independently a single bond, F double bond, Z double bond, or triple bond,
`
`as valency permits; and
`
`n is an integer between 6 and 20, inclusive.
`
`In some embodiments, the lactone comprises one or more compoundsof the formula:
`
`10
`
` ~wherein:
`
`R is methyl, ethyl, or n-propyl;
`
`each --- is independently a single bond or Z double bond,as valency permits, wherein
`
`0, 1,2, or 4 --- are Z double bonds; and
`
`m is an integer between 4 and 11, inclusive.
`
`15
`
`In some embodiments, the lactone comprises one or more compoundsselected from:
`
`O
`
`O
`
`o7
`
`So
`
`Oo”
`
`“o
`
`Oo
`
`O
`
`Oo
`
`©
`
`QO
`
`oO
`
`O
`
`0
`
`oO”
`
`“o
`
`SS
`
`SK
`
`ww
`
`oae)
`
`

`

`
`
`

`

`
`
`In some embodiments, the lactone comprises one or more compoundsselected from:
`
`

`

`
`
`In some embodiments, the lactone comprises one or more compoundsselected from:
`
`

`

` , and
`
`

`

`
`
` and
`
`In some embodiments, the chiral carbon atom is of the S configuration. In some
`
`embodiments, the chiral carbon atom is of the R configuration.
`
`In some cmbodiments, the consumer product is for skin application. In some
`
`embodiments, the consumer product is for non-skin application. In some embodiments, the
`
`10
`
`consumerproduct is a perfume product. In some embodiments, the consumerproductis a skin
`
`care product. In some embodiments, the consumer productis a hair care product. In some
`
`embodiments, the consumer product is a feminine hygiene product. In some embodiments, the
`
`consumer product is a fabric care product. In some embodiments, the consumer productis a
`
`cleaning product. In some embodiments, the consumer product further comprises a second
`
`

`

`9
`
`fragrance. In some embodiments, the lactone is a booster for the second fragrance. In some
`
`embodiments, the lactone boosts overall fragrance performance. In some embodiments, the
`
`consumerproductis for medicinal use, optionally wherein the medicinal use is anti-
`
`inflammatory. In some embodiments, the consumer product is for improving mood. In some
`
`embodiments, the lactone has increased substantivity. In some embodiments, the lactone has
`
`increased biodegradability. In some embodiments, the consumer product is selected from the
`
`group consisting of: fragrance, body wash, shampoo, after bath splash, eau de toilette, cologne,
`
`lotion, cream, liquid laundry detergent, compressed cleaning tablet, lip gloss, solid body
`
`moisturizer bar, hair care mousse, scented ink, gel hand sanitizer, candle, all-purpose cleaner,
`
`10
`
`linen spray, fabric softener, dishwashing liquid, deodorantstick, soap, scented garbage bags,
`
`perfume bearing microcapsules, and eye patch.
`
`In some embodiments, the lactone is produced by a method comprising:
`
`(i) preparing a first reaction mixture comprising one or more fatty acids, a cytochrome
`
`P450 hydroxylase, and NADPH;
`
`15
`
`20
`
`(ii) incubating the first reaction mixture of for a sufficient time to produce hydroxyl
`
`fatty acids selected from -1 hydroxyl fatty acids, w-2 hydroxyl fatty acid, w-3 hydroxyl fatty
`
`acid, and combinationsthereof;
`
`(iii) preparing a second reaction mixture comprising the hydroxyl fatty acids produced
`
`in step (ii) and a lipase; and
`
`(iv) incubating the second reaction mixture for a sufficient time to produce the lactone.
`
`25.
`
`The method of claim 24, wherein step (i1) further comprises isolating the hydroxy] fatty
`
`acids from thefirst reaction mixture.
`
`In some embodiments, the cytochrome P450 hydroxylase comprises an amino acid
`
`sequencethat is at least 80% identical to the amino acid sequence of SEQ ID NO: 1 or SEQ ID
`
`25
`
`NO: 3. In some embodiments, the cytochrome P450 hydroxylase comprises the amino acid
`
`sequence of SEQ ID NO: 1 or SEQ ID NO: 3. In some embodiments, the lipase is lipase B
`
`from Candida antarctica. In some embodiments, the lipase comprises an amino acid sequence
`
`that is at least 80% identical to the amino acid sequence of SEQ ID NO: 5. In some
`
`embodiments, the lipase comprises the amino acid sequence of SEQ ID NO: 5. In some
`
`30
`
`embodiments, the lipase is immobilized on a solid support. In some embodiments, the second
`
`reaction mixture further comprises a solvent, optionally wherein the solvent is toluene or
`
`dichloroethane. In some embodiments, the hydroxyl fatty acids in the second reaction mixture
`
`

`

`10
`
`are at a total concentration of 0.02-0.1 M, optionally wherein the hydroxyl fatty acids are at a
`
`total concentration of 0.025-0.5 M. In some embodiments, the lipase in the second reaction
`
`mixture is at a concentration of 20-150 g/L, optionally wherein the lipase is at a concentration
`
`of 50-100 g/L. In some embodiments, step (iv) further comprises isolating the lactone.
`
`In some embodiments, the one or more fatty acids of step (4) comprise a linear fatty
`
`acid comprising 12-28 carbon atoms, optionally wherein the one or morefatty acids of step (i)
`
`comprisc a linear fatty acid comprising 15, 16, 17, 18, or 20 carbon atoms. In some
`
`embodiments, the one or more fatty acids of step (4) comprise a saturated fatty acid. In some
`
`embodiments, the one or more fatty acids of step (1) comprise an unsaturated fatty acid,
`
`10
`
`optionally wherein the unsaturated fatty acid comprises at least one double bond, optionally
`
`wherein the unsaturated fatty acid comprisesat least one Z double bond.
`
`In some embodiments, the one or morefatty acids of step (i) are selected from the
`
`group consisting of:
`
`15
`
`20
`
`ot (C15:0, pentadecanoic acid);
`
`O
`
`OEE (C16:0, palmitic acid);
`
`oO
`
`Oo
`
`(C16:1, palmitoleic acid);
`
`eee (C17:0, heptadecanoicacid);
`
`Oo
`
`NEE (C18:0, octadecanoic acid);
`
`Oo
`
`IO (C18:1, oleic acid);
`
`0
`
`

`

`5
`
`11
`
`eeeaS (C18:2, linoleic acid);
`
`oO
`
`O
`
`O
`
`OH (C20:4, arachidonic acid);
`
`OH ((7Z,102Z,137)-Hexadecatrienoic acid);
`
`oO
`
`oO
`
`OH ((6Z,9Z,12'Z)-Hexadecatrienoic acid);
`
`S OSS
`
`OH ((4E,7E,10E)-Hexadecatricnoic acid);
`
`9
`
`LOT “OH ((2E,4E,6E)-Hexadecatrienoic acid);
`
`oO
`NN
`
`oH (y-linolenic acid);
`
`9
`
`oH (a-linolenic acid);
`
`oO
`
`10VARReet ((8E,10E,127)-Octadecatrienoic acid);
`
`9
`
`RRRht
`
`((9Z,11E,13E)-octadeca-9,11,13-trienoic acid);
`
`OP ((9QE, 1 1E,13E)-octadeca-9,11,13-tricnoic acid);
`
`Oo
`
` ((5Z,9Z, 12Z)-octadeca-5,9,12-tricnoic acid);
`
`=
`
`OH
`
`Oo
`
`= —
`
`(8Z,11Z,147Z-eicosatrienoic acid);
`
`

`

`12
`
`S
`
`((5Z,8Z, 1 1Z)-Eicosa-5,8,11-trienoic acid);
`
`and combinations thereof.
`
`In some embodiments, the one or more fatty acids of step G1) comprise
`
`Rt (C15:0, pentadecanoic acid) and the lactone
`
`O
`
`produced in step (iv) comprises:
`
`combinations thereof.
`
`oO
`
`O
`
`PT
`
`NS
`
`,
`
`07
`
`So
`
`,
`
`, and
`
`In some embodiments, the one or more fatty acids of step (i) comprise
`
`oO
`ONE
`
`OH (C16:0, palmitic acid) and the lactone produced in
`
`O
`
`O
`
`O
`
`O
`
`O
`
`step (iv) comprises:
`
`;
`
`>
`
`, and
`
`10
`
`combinations thereof.
`
`In some embodiments, the one or more fatty acids of step (i) comprise
`
`a
`
`O
`
`OH
`
`(C16:1, palmitoleic acid) and the lactone produced in
`
`

`

`13
`
`O
`
`O
`
`O
`
`step (iv) comprises: S
`
`; S
`
`> ~
`
`, and
`
`combinations thereof.
`
`In some embodiments, the one or more fatty acids of step G1) comprise
`O
`
`Wt (C17:0, heptadecanoic acid) and the lactone
`
`O
`
`O._0
`
`O
`
`oO
`
`producedin step (iv) comprises:
`
`,
`
`,
`
`O
`
`, and
`
`combinations thereof.
`
`In some embodiments, the one or more fatty acids of step (i) comprise
`Oo
`NNNEEaEeaeeea
`OH (C18:0, octadecanoic acid) and the lactone
`
`O
`
`O
`
`O
`
`oO
`
`O
`
`produced in step (iv) comprises:
`
`;
`
`>
`
`O
`
`>
`
`10
`
`and combinations thereof.
`
`In some embodiments, the one or more fatty acids of step (i) comprise
`
`O
`
`—IO (C18:1, oleic acid) and the lactone produced in
`
`step(iv)comprises:oyaouandcombinations
`
`thereof.
`
`

`

`14
`
`In some embodiments, the one or more fatty acids of step (i) comprise
`
`NEEOH (C 18:2, linoleic acid) and the lactone produced
`
`
`
`,
`
`, and
`
`in step (iv) comprises:
`
`combinations thereof.
`
`In some embodiments, the one or more fatty acids of step (i) comprise
`
`0
`
`OH (C20:4, arachidonic acid) and the
`
`
` , and combinations thereof.
`
`In some embodiments, the one or more fatty acids used as substrates in the first
`9
`
`10
`
`reaction mixture in step (i) compriseNNN
`( (7Z,10Z,13Z)-Hexadecatrienoic acid) and the lactone produced in step Gv) comprises:
`
`
`
`In some embodiments, the one or more fatty acids used as substrates in the first
`9
`
`reaction mixture in step (1) comprise
`
`— = —=
`
`OH ((6Z,9Z, 12Z,)-
`
`

`

`
`
` , and combinations thereof.
`
`In some embodiments, the one or more fatty acids used as substrates in thefirst
`°
`NNRNee
`reaction mixture in step (1) comprise
`aaa oH ((4E,7E,10E)-
`
`
`
` , and combinations thereof.
`
`In some embodiments, the one or more fatty acids used as substrates in the first
`9
`
`reaction mixture in stcp (i) comprisc eeeOH ((2E,4E,6E)-
`
`O
`
`= O
`\\
`
`<
`
` ;
`
`, and combinations thereof.
`
`5
`
`10
`
`In some embodiments, the one or more fatty acids used as substrates in thefirst
`
`Aeeeeet (y-linolenic acid) andreaction mixture in step (1) comprise
`
`
`
`

`

`16
`
` the lactone produced in step (iv) comprises:
`
`and combinations thereof.
`
`In some embodiments, the one or more fatty acids used as substrates in the first
`Oo
`
`reaction mixture in step (i) comprise
`
`OH (a-linolenic acid) and
`
`O
`
`O
`
`the lactone producedin step (iv) comprises:
`
`SS
`
`a
`
`In some embodiments, the one or more fatty acids used as substrates in thefirst
`0
`
`reaction mixture in step (i) compriseeat (a-Calendic acid;
`(8E,10E,12Z)-Octadecatrienoic acid) and the lactone producedin step (iv) comprises:
`
`oOmSCoeandcombinationsthereof.
`
`In some embodiments, the one or more fatty acids used as substratesin the first
`0
`
`reaction mixture in step (i) compriseRt ((9Z,11E,13E)-
`octadeca-9,11,13-trienoic acid) and the lactone producedin step (iv) comprises:
`
`O
`
`Oo
`
`
`
`oO
`
`—ee
`
`|
`|
`SS ; a . and combinations thereof.
`
`In some embodiments, the one or more fatty acids used as substrates in the first
`Oo
`
`15
`
`reaction mixture in step (i) compriseABO ((9E,11E,13E)-
`
`

`

`octadeca-9,11,13-trienoic acid) and the lactone produced in step (iv) comprises:
`
`17
`
`O
`
`O
`
`Vy,
`
`,
`
`, and combinations thereof.
`
`In some embodiments,
`
`the one or more fatty acids used as substrates in thefirst
`
`
`Oo
`-
`Colo
`-
` reaction mixture in step (1) comprise
`
`
` VN
`OH reaction mixture in step (1) comprise
`
`
`In some embodiments, the one or more fatty acids used as substrates in the first
`
`, and combinations thereof.
`
`(8Z,11Z,14Z-eicosatrienoic
`
` , and combinations thereof.
`
`10
`
`

`

`In some embodiments, the one or more fatty acids used as substrates in the first
`
`18
`
`O
`
`a
`
`OH
`
`reaction mixture in step (1) comprise S
`
` , and combinations thereof.
`
`((5Z,8Z, 11Z)-Eicosa-5,8,11-
`
`
`
`In some embodiments, the first reaction mixture is in vitro. In some embodiments, the
`
`first reaction is a cell-based reaction mixture. In some embodiments, the cell-based reaction
`
`mixture comprises a cell selected from the group consisting of a yeast, a plant, an alga, a
`
`fungus, and a bacterium. In some embodiments, the cell-based reaction mixture comprises a
`
`bacterial cell of a genus selected from the group consisting of Escherichia; Salmonella;
`
`10
`
`Bacillus; Acinetobacter; Corynebacterium; Methylosinus; Methylomonas; Rhodococcus;
`
`Pseudomonas; Rhodobacter; Synechocystis; Brevibacteria; Microbacterium; Arthrobacter;
`
`Citrobacter; Escherichia; Klebsiella; Pantoea; Salmonella; Corynebacterium, and
`
`Clostridium, optionally wherein the cell-based reaction mixture comprises an EF. coli cell. In
`
`some embodiments, the cell-based reaction mixture comprises a fungus of a genus selected
`
`15
`
`from the group consisting of Saccharomyces; Zygosaccharomyces,; Kluyveromyces; Candida;
`
`Streptomyces; Hansenula; Debaryomyces; Mucor; Pichia; Torulopsis; Aspergillus; and
`
`Arthrobotlys.
`
`In some embodiments, the lactone producedin step (iv) has a purity of at least 70%.
`
`Other aspects of the present disclosure provide methods of producing a relaxed mood in
`
`20
`
`a subject in need of same, the method comprising administering to the subject an effective
`
`amount of a fragrance composition described herein. Other aspects of the present disclosure
`
`provide method of reduce inflammation in a subject in need of same, the method comprising
`
`administering to the subject an effective amountof a fragrance composition described herein.
`
`In some embodiments, the subject is human.
`
`

`

`19
`
`Otheraspects of the present disclosure provide methods of improving, enhancing, or
`
`modifying the fragrance of a fragrant composition, the method comprising adding a lactone
`
`comprising one or more compoundsof the formula:
`
`wherein:
`
`R is methyl, ethyl, or n-propyl;
`
`each --- is independently a single bond, F double bond, Z double bond, or triple bond,
`
`as valency permits; and
`
`k is an integer between 6 and 30, inclusive;
`
`10
`
`In some embodiments, the method comprises adding a lactone comprising one or more
`
`compounds of the formula:
`
`0
`
`aso
`(QePR.
`
`wherein:
`
`R is methyl, ethyl, or n-propyl;
`
`15
`
`each --- is independently a single bond, F double bond, Z double bond, or triple bond,
`
`as valency permits; and
`
`nis an integer between 6 and 20, inclusive.
`
`In some embodiments, the lactone comprises one or more compoundsselected from:
`
`

`

`
`
`

`

`21
`
`

`

`22
`
`In some embodiments, the lactone comprises one or more compoundsselected from:
`
`
`CREAR
`PePEER
`EBLOS
`
`
`
`

`

`In some embodiments, the lactone comprises one or more compoundsselected from:
`
`O
`
`O
`
`O
`
`0
`
`SS
`
`SA
`
`SN
`
`O
`
`Oo
`
`Oo
`
`O
`
`O
`
`O
`
`O
`
`oO
`
`O
`
`O
`
`O
`
`SS
`
`SS
`
`O
`
`—
`
`O
`
`

`

`24
`
` and
`
`While the disclosure is susceptible to various modifications and alternative forms,
`
`specific embodiments thereof are shown by way of example in the drawing and will herein be
`
`described in detail. It should be understood, however, that the drawings and detailed
`
`description presented herein are not intended to limit the disclosure to the particular
`
`10
`
`embodiments disclosed, but on the contrary, the intention is to cover all modifications,
`
`equivalents, and alternatives falling within the spirit and scope of the present disclosure as
`
`defined by the appended claims.
`
`

`

`25
`
`Other features and advantages of this invention will become apparent in the following
`
`detailed description of preferred embodiments of this invention, taken with reference to the
`
`accompanying drawings.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`The accompanying drawings are not intended to be drawnto scale. In the drawings,
`
`each identical or nearly identical componentthat is illustrated in various figures is represented
`
`by a like numeral. For purposes of clarity, not every component may be labeled in every
`
`drawing. In the drawings:
`
`10
`
`15
`
`20
`
`FIG. 1. Overall synthetic scheme showing fatty acid hydroxylation (Process 1) and a
`
`reaction of obtaining macrolactones (Process 2).
`
`FIG. 2. GC/MSanalysis of reaction products catalyzed by lipase. The numbersindicate
`
`the musk lactone peaks from different fatty acids. The molecular weights of these musky
`
`lactones match their calculated molecular weights.
`
`FIGs. 3A-3F. Reaction schemes showing fatty acid hydroxylation and ring closure for
`
`oleic acid (FIG. 3A), linoleic acid (FIG. 3B), arachidonic acid (FIG. 3C), palmitic acid (FIG.
`
`3D), heptadecanoic acid (FIG. 3E), and octadecanoic acid (FIG. 3P).
`
`FIG. 4. Overall synthetic scheme showing palmitic acid hydroxylation (Process 1) and
`
`a reaction of obtaining corresponding musk lactone products (Process 2).
`
`FIG. 5. Reaction showing hydroxy] oleic acid cyclized to obtain corresponding musk
`
`lactone products (Process 2).
`
`FIG. 6. Chromatogram and spectrum showing analysis of the oleic acid
`
`macrolactonization products.
`
`FIG. 7. Overall synthetic scheme showing linoleic acid hydroxylation (Process 1) and a
`
`25
`
`reaction of obtaining corresponding musk lactone products (Process 2) is illustrated in FIG. 6.
`
`FIG. 8. GC/MSanalysis of the linoleic acid macrolactonization products.
`
`FIG. 9. Production of musk lactones from hydroxy y-linolenic acid (GLA, C18:3).
`
`FIG. 10. GC/MSanalysis of musk lactones derived from hydroxy GLA (molecular
`
`weight: 276).
`
`30
`
`FIG. 11. Production of musky lactone from a-linolenic acid (ALA, C18:3).
`
`FIG. 12. GC/MSanalysis of musk lactone from hydroxy ALA (molecular weight: 276).
`
`FIG. 13. Production of musk lactone from 8Z,112Z,14Z-eicosatrienoic acid, dihomo-y-
`
`linolenic acid (DGLA, C20:3).
`
`

`

`FIG. 14. GC/MSanalysis of musk lactone from hydroxy DGLA (molecular weight:
`
`304).
`
`26
`
`DEFINITIONS
`
`The term "about" or "approximately" means within an acceptable error range for the
`
`particular value as determined by one of ordinary skill in the art, which will depend in part on
`
`how the value is measured or determined,i.e., the limitations of the measurement system. For
`
`example, "about" can mean within 3 or more than 3 standard deviations, per the practice in the
`
`art. Alternatively, “about” can meana range of up to 20%, preferably up to 10%, more
`
`10
`
`preferably up to 5%, and more preferably still up to 1% of a given value. Alternatively,
`
`particularly with respect to biological systems or processes, the term can mean within an order
`
`of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value.
`
`As used herein, the term "weight percent" or "wt-%" is meant to refer to the quantity by
`
`weight of a componentin a material as a percentageof the total wet weight of the material (i.e.,
`
`15
`
`a fragrance formulation). Unless otherwise specified, all amounts expressed as a percentage
`
`herein represent the amount in weight percent.
`
`The terms "fragrance composition", "fragrance", "fragrance formulation", "perfume"
`
`and "perfume composition" meanthe sameto refer to a perfumed compositionthat is a mixture
`
`of fragrance compounds including for example, alcohols, aldehydes, ketones, esters, ethers,
`
`20
`
`acetates, nitriles, terpene hydrocarbons, nitrogenous or sulphurous heterocyclic compounds and
`
`essential oils of natural or synthetic origin, which are admixed so that the combined odors of
`
`the individual components produce a pleasant or desired fragrance. Manyof these ingredients
`
`are listed in reference texts such as S. Arctander, Perfume and Flavor Chemicals, 1969,
`
`Montclair, N.J., USA (or any of its more recent versions), which is herein incorporated by
`
`25
`
`reference in its entirety. As described herein, fragrance compositions can be a mixture of any
`
`number of fragrance compounds. For example, fragrance compositions include "simple
`
`accords", e.g., having fewer than 10 fragrance compounds, and "complex fragrances", e.g.,
`
`having more than 30 fragrance compounds. In certain embodiments, the fragrance
`
`compositions of the present disclosure can be a combination of 2 or more accords.
`
`30
`
`As used herein, the term "improving" is understood to mean raising a fragrance
`
`composition to a more desirable character, the term "enhancing" is understood to mean making
`
`the fragrance composition greater in effectiveness, such as strength, and the term "modifying"
`
`is understood to mean providing the fragrance composition with a change in character.
`
`

`

`27
`
`As used herein, the term "isomers" refers to different compoundsthat have the same
`
`molecular formula but differ in arrangement and configuration of the atoms. Also as used
`
`herein, the term "stereoisomer”refers to any of the various stereo isomeric configurations
`
`which mayexist for a given compoundof the presently disclosed subject matter and includes
`
`geometric isomers. It is understood that a substituent may be attached at a chiral center of a
`
`carbon atom. Therefore, the presently disclosed subject matter includes enantiomers,
`
`diastereomers or racemates of the compound. Also as used herein, the terms "constitutional
`
`isomers" refers to different compounds which have the same numbersof, and types of, atoms
`
`but the atoms are connected differently.
`
`10
`
`As used herein, the term "diastereoisomers" refers to stereoisomers that have at least
`
`two asymmetric atoms, but which are not mirror-images of each other. The absolute
`
`stereochemistry is specified according to the Cahn-Ingold- Prelog R-S system. When a
`
`compoundis a pure enantiomer the stereochemistry at each chiral carbon may be specified by
`
`either R or S. Resolved compounds whose absolute configuration is unknown can be
`
`15
`
`designated (+) or (-) depending on the direction (dextro or levorotatory) which they rotate
`
`plane polarized light at the wavelength of the sodium D linc. The compoundsof the presently
`
`disclosed subject matter contain one or more asymmetric centers and maythus giverise to
`
`enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of
`
`absolute stereochemistry, as (R)- or (S)-. The presently disclosed subject matter is meant to
`
`20
`
`include all such possible isomers, including racemic mixtures, optically pure forms and
`
`intermediate mixtures. Optically active (R)- and (S)- isomers may be prepared using chiral
`
`synthonsor chiral reagents, or resolved using conventional techniques. If the compound
`
`contains a double bond, the substituent may be E or Z configuration. If the compound contains
`
`a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis- or trans-configuration. All
`
`25
`
`tautomcric forms arc also intended to be included.
`
`As used herein, the term "enantiomers" refers to a pair of stereoisomers that are non-
`
`superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a
`
`"racemic" mixture. The term is used to designate a racemic mixture where appropriate.
`
`The term “alkyl” refers to a radical of a branched or unbranched,saturated acyclic
`
`30
`
`hydrocarbon group. In certain embodiments, alkyl is C3-36 alkyl. In certain embodiments, alkyl
`
`is Cio-36 alkyl. In certain embodiments, alkyl is Ci1-27 alkyl. Unless otherwise provided, alkyl is
`
`C329 alkyl.
`
`

`

`28
`
`The term “alkenyl” refers to a radical of a branched or unbranched, acyclic
`
`hydrocarbon group having one or more carbon-carbon double bonds (C=C bonds; e.g., 1, 2, 3,
`
`hy
`
`4,5, or 6 C=C bonds), as valency permits. In alkenyl groups, “+
`is an E double bond,
`“ M is an Z double bond. Other situations involving an £ or Z double bond are as known
`
`in the art. In an alkenyl group, a C=C bond for which the stereochemistry is not specified (e.g.,
`
`a—CH=CH-or
`
`) may be a F or Z double bond. In certain embodiments, alkenyl is C3.
`
`36 alkenyl. In certain embodiments, alkenyl is Cio-36 alkenyl. In certain embodiments, alkenyl]is
`
`Ci1-27 alkenyl. Unless otherwise provided, alkenyl is C329 alkenyl.
`
`The term “alkynyl” refers to a radical of a branched or unbranched, acyclic
`
`hydrocarbon group having one or more carbon-carbon triple bonds (C=C bonds; e.g., 1, 2, 3, or
`
`4 triple bonds), as valency permits. In certain embodiments, alkynyl is C3-36 alkynyl. In certain
`
`embodiments, alkynyl is Cio-36 alkynyl. In certain embodiments, alkynyl is C11-27 alkynyl.
`
`Unless otherwise provided, alkynyl is C329 alkynyl.
`
`Affixing the suffix “ene” to a group indicates the group is a divalent moiety, e.g.,
`
`alkyleneis a divalent moiety of alkyl (e.g., Cs-36 alkyl, Cio-36 alkyl, Ci1-27 alkyl, or C3-29 alkyl),
`
`alkenyleneis a divalent moiety of alkenyl(e.g., C3-36 alkenyl, Cio-36 alkenyl, C11-27 alkenyl, or
`
`C329 alkenyl), and alkynyleneis a divalent moiety of alkynyl (e.g., C336 alkynyl, C10-36
`
`alkynyl, C11-27 alkynyl, or C3-29 alkynyl).
`A “fatty acid”is a carboxylic acid of the formula: R¢-C(=O)OH,whercin R4 is C3-36
`
`alkyl, C3-36 alkenyl, or C3-36 alkynyl (e.g., C3-29 alkyl, C3-29 alkenyl, or C329 alkynyl). The
`carbon atom (e.g., C1) in R“ that is farthest to the carboxyl moiety is labelled as m (omega).
`
`The carbon atom(e.g., C2) next to C1 is labelled as w—1. The carbon atom(e.g., C3) that is
`
`next to C2 and is not Cl is labelled as w—2. The carbon atom (e.g., C4) that is next to C3 and is
`
`not C2 is labelled as m—3. A 15:0 fatty acid is a fatty acid where the numberof carbon atomsis
`
`10
`
`15
`
`20
`
`25
`
`15, and the number of C=C and C=C bondsis 0. A 16:0 fatty acid is a fatty acid where the
`
`number of carbon atoms is 16, and the number of C=C and C=C bondsis 0. A 16:1 fatty acid is
`
`a fatty acid where the numberof carbon atomsis 16, the number of C=C bondsis 1, and the
`
`number of C=C bondsis 0. A 16:3 fatty acid is a fatty acid where the numberof carbon atoms
`
`is 16, the number of C=C bondsis 3, and the number of C=C bondsis 0. A 17:0 fatty acid is a
`
`30
`
`fatty acid where the numberof carbon atomsis 17, and the number of C=C and C=C bondsis
`
`0. An 18:0 fatty acid is a fatty acid where the number of carbon atomsis 18, and the numberof
`
`

`

`29
`
`C=C and C=C bondsis 0. An 18:1 fatty acid is a fatty acid where the numberof carbon atoms
`
`is 18, the number of C=C bondsis 1, and the number of C=C bondsis 0. An 18:2 fatty acid is a
`
`fatty acid where the numberof carbon atomsis 18, the number of C=C bondsis 2, and the
`
`number of C=C bondsis 0. An 18:3 fatty acid is a fatty acid where the number of carbon atoms
`
`is 18, the number of C=C bondsis 3, and the number of C=C bondsis 0. A 20:3 fatty acid is a
`
`fatty acid where the numberof carbon atomsis 20, the number of C=C bondsis 3, and the
`
`number of C=C bondsis 0. A 20:4 fatty acid is a fatty acid where the numberof carbon atoms
`
`is 20, the number of C=C bondsis 4, and the number of C=C bondsis 0. Other fatty acids can
`
`be namedin this manner.
`
`10
`
`A “saturated fatty acid”is a fatty acid where R“ is C336 alkyl (e.g., C329 alkyl).
`An “unsaturated fatty acid” is a fatty acid where R“ is C3-36 alkenyl or C3-36 alkynyl
`
`(e.g., C3-29 alkenyl or C3-29 alkynyl).
`
`The term “hydroxyl” or “hydroxy” refers to the group —OH.
`
`A “hydroxyl fatty acid,” “hydroxy fatty acid,” or “hydroxylated fatty acid” is a fatty
`
`15
`
`acid where one or more hydrogen atom is replaced with hydroxyl. In some embodiments, the
`
`hydroxyl fatty acid is a fatty acid where one hydrogen atom is replaced with hydroxyl (mono-
`
`hydroxyl fatty acid).
`
`In some embodiments, the hydroxyl fatty acid is a fatty acid where more
`
`than one (e.g., 2, 3, or more) hydrogen atom is replaced with hydroxyl (poly-hydroxy] fatty
`
`acid).
`
`20
`
`A “lactone” is a monocyclic compound where the moiety —C(=O)O-is part of the
`
`monocyclic ring, and the remaining part of the monocyclic compoundis alkylene, alkenylene,
`
`or alkynylene. When the alkylene, alkenylene, or alkynylene is branched, the lactone also
`
`includesthe branch(es) of the alkylene, alkenylene, or alkynylene.
`
`“Cellular system” is any cells that provide for the expression of ectopic protcins. It
`
`25
`
`includes bacteria, yeast, plant cells and animalcells. It may include prokaryotic or eukaryotic
`
`host cells which are modified to express a recombinant protein and cultivated in an appropriate
`
`culture medium. It also includes the in vitro expression of proteins based on cellular
`
`components, such as ribosomes.
`
`"Coding sequence"is to be given its ordinary and customary meaning to a person of
`
`30
`
`ordinary skill in the art, and is used without limitation to refer to a DNA sequence that encodes
`
`for a specific amino acid sequence.
`
`“Growing the Cellular System’’. Growing includes providing an appropriate medium
`
`that would allow cclls to multiply and divide, to form a cell culture. It also includes providing
`
`resources so that cells or cellular components can translate and make recombinant proteins.
`
`

`

`30
`
`“Protein Expression”. Protein production can occurafter gene expression. It consists of
`
`the stages after DNA has been transcribed to messenger RNA (mRNA). The mRNAis then
`
`translated into polypeptide chains, which are ultimately folded into proteins. DNA or RNA
`
`may be presentin the cells through transfection - a process of deliberately introducing nucleic
`
`acids into cells. The term is often used for non-viral methods in eukaryotic cells. It may also
`
`refer to other methods and cell types, although other termsare preferred: "transformation"is
`
`more often used to describe non-viral DNAtransfer in bacteria, non-animal eukaryotic cells,
`
`including plant cells. In animal cells, transfection is the preferred term as transformation is
`
`also used to refer to progression to a cancerous state (carcinogenesis) in these cells.
`
`10
`
`Transduction is often used to describe virus-mediated DNAtransfer. Transformation,
`
`transduction, and viral infection are included underthe definition of transfection for this
`
`application.
`
`“Yeast”. According to the current disclosure a yeast are eukaryotic, single-celled
`
`microorganismsclassified as members of the fungus kingdom. Yeasts are unicellular
`
`15
`
`organisms whichare believed to have evolved from multicellular ancestors.
`
`As used herein, the singular forms "a, an" and "the" include plural references unless the
`
`content clearly dictates otherwise.
`
`To the extent that the term "include," "have," or the like is used in the description or the
`
`claims, such term is intended to be inclusive in a manner similar to the term "comprise" as
`
`20
`
`“comprise” is interpreted when employed as a transitional wordin a claim.
`
`The word "exemplary" is used herein to mean "serving as an example, instance, or
`
`illustration.” Any embodiment described herein as “exemplary” is not necessarily to be
`
`construed as preferred or advantageous over other embodiments.
`
`The term “complementary” is to be given its ordinary and customary meaning to a
`
`25
`
`person of ordinary skill in the art and is used without limitation to describe the relationship
`
`between nucleotide bases that are capable to hybridizing to one another. For example, with
`
`respect to DNA, adenosine is complementary to thymine and cytosine is complementary to
`
`guanine. Accordingly, the subject technology also includesisolated nucleic acid fragments
`
`that are complementary to the complete sequences as reported in the accompa