Improvements in or relating to organic compounds

Abstract

A flavor mixture comprising a compound according to formula (I) or formula (II) and a method of providing such mixture. Formula I Formula II

Description

Technical Field

[0001] This invention generally relates to novel flavoring mixtures, methods for preparing them, and their use in consumer products.

[0002] sequence list

[0003] This application contains a sequence list, which is electronically submitted in XML format conforming to WIPO Standard ST.26 and is incorporated herein by reference in its entirety. The XML copy was created on November 18, 2025, named 31536.xml, and has a size of 13471 bytes. Background of the Invention

[0005] Fragrance components that deliver the scent profile of ambergris in consumer products are essential materials in a perfumer's toolbox. They add depth and warmth to fragrances, and are therefore often used as base fragrances in the fragrance industry to suggest feelings of warmth, sophistication, luxury, and elegance.

[0006] Ambermax TM Fragrance ingredient is well-known in the fragrance industry. When used in fine and functional fragrances, it delivers a strong, blended, and long-lasting rich ambergris scent with hints of woody cypress. Its exceptional fabric retention makes it a key ingredient in fabric care fragrances, outperforming all benchmarks on dry fabrics. It is widely used in home care, laundry care, and personal care products, as well as in the fine fragrance industry, to produce pleasant scents or, if needed, to mask unpleasant odors.

[0007] Ambermax TM The flavoring ingredient was first described in U.S. Patent 7,550,417. Even a cursory examination of the molecule's structure is sufficient for a person with relevant expertise to identify its multiple stereocenters and positional isomers. In fact, the ingredient exists commercially as a fine mixture of its stereoisomers, regioisomers, structural isomers, and conformational isomers.

[0008] For modern spice producers focused on the sustainable use of precious resources, Ambermax TM Fragrance components often exist as complex mixtures of isomers, which presents a problem: only some of these isomers may contribute significantly to the overall aroma of the component, and some isomers may even produce unpleasant odors.

[0009] There is still a need to provide new spice blends that can impart the commercial ingredient Ambermax TM The aromas are the same or substantially similar, but this blend represents a more efficient use of carbon. Invention Overview

[0011] To address the challenge of providing more sustainable utilization of fragrance components, the applicant employed a more discriminative and sensitive olfactory receptor assay technique to study the activity of fragrance components at the receptor level, thereby gaining a clearer understanding of the contribution of various isomers of complex fragrance components to their odor characteristics and intensity. In fact, in the fragrance component Ambermax... TM In this case, the applicant found that only a very small number of isomers contributed to the properties and strength of the ingredient, while other isomers had only a faint odor or even exhibited an undesirable off-odor. This insight enabled the applicant to propose a product rich in commercial Ambermax. TM A new fragrance blend of isomers that strongly contribute to the odor characteristics and intensity of the fragrance components, but which contains or substantially contains no isomers that do not contribute to or are harmful to the desired odor characteristics and intensity, and also contains or substantially contains no such byproducts.

[0012] Therefore, in a first aspect of the invention, a fragrance mixture is provided comprising compounds according to Formula I and Formula II.

[0013]

[0014] Formula I Formula II

[0015] The spice mixture contained no or substantially no compounds according to Formula III.

[0016]

[0017] Formula III,

[0018] Furthermore, in the compounds of formulas I, II, and / or III, the wavy bonds represent unspecified configurations at the connected carbon atoms, and in the compound of formula III, the dashed lines are combined with the solid lines ( ) represents a single bond and a double bond.

[0019] In a second aspect of the invention, a fragrance composition is provided comprising the fragrance mixture of the first aspect of the invention and at least one other fragrance ingredient.

[0020] In a third aspect of the invention, a consumer product is provided comprising a fragrance mixture according to a first aspect of the invention.

[0021] In a fourth aspect of the invention, a method for preparing a spice mixture according to the first aspect of the invention is provided.

[0022] Details, embodiments, and preferred embodiments provided with respect to any one or more of the described aspects of the invention will be further described herein, and are equally applicable to all aspects of the invention. Unless otherwise stated herein, or clearly contradicted by the context, the invention covers any combination of all possible variations of the embodiments, embodiments, and preferred embodiments described below. Invention Details

[0024] A first aspect of the invention relates to a fragrance mixture comprising compounds according to Formula I and Formula II.

[0025]

[0026] Formula I Formula II

[0027] The spice mixture described herein contains no or substantially no compounds according to formula IIIa and / or IIIb.

[0028]

[0029] Formula IIIa Formula IIIb

[0030] Furthermore, the wavy bonds in the compounds of formulas I, II, IIIa and / or IIIb represent unspecified configurations at the connected carbon atoms.

[0031] To the best of the applicant's knowledge, the preparation and characterization of the isomer mixture and its use in the fragrance industry are unknown in the art.

[0032] As assessed by a specialized team of trained members with normal olfactory sensitivity, the fragrance blends, including all specific embodiments described below, were found to have commercial-grade Ambermax. TM The ingredients have desirable odor properties; however, compared to commercial-grade ingredients, the fragrance blends exhibit more efficient carbon utilization because they are rich in certain desired isomers and contain little or no isomers that lack essential odor characteristics or intensity.

[0033] In a specific embodiment of the invention, a fragrance mixture comprising a compound according to formula I and a compound according to formula II is provided, wherein the ratio of the compound of formula (I) to the compound of formula (II) is 1.3 to 2.5, or 1.5 to 2.0, or 1.7 to 1.9.

[0034] In a particular embodiment of the invention, the spice mixture comprises 50-70% by weight of a compound according to formula I and 28-38% by weight of a compound according to formula II.

[0035] As used herein, the term “substantially free” means that, in terms of the extent to which the isomer is present in the flavor blend, it is present in trace amounts below the levels commonly used in the flavor industry, as practiced by those skilled in the art.

[0036] Specifically, the applicant discovered that the ingredient Ambermax, as reported in US7,550,417, contains... TM The tetracyclic ethers of formula IIIa and / or IIIb present in amounts up to 20 wt% do not contribute significantly or at all to the odor characteristics or intensity of the component.

[0037] More specifically, its presence will be less than 0.1 wt%, based on the total weight of the spice mixture.

[0038] In embodiments of the invention, if present, the tetracyclic ethers of formula IIIa and / or IIIb are present at a level of 0.1 wt% or less.

[0039] In a specific embodiment of the present invention, a flavoring mixture is provided comprising 50-70% by weight of a compound according to formula I, 28-38% by weight of a compound according to formula II, 0.1 wt% or less of a compound according to formula IIIa, and 0.1 wt% or less of a compound according to formula IIIb.

[0040] In one specific embodiment of the invention, the fragrance mixture is free of or substantially free of byproducts that do not contribute to or interfere with the desired olfactory characteristics or intensity of the fragrance mixture. More specifically, the fragrance mixture is free of or substantially free of compounds according to Formula IV or Formula V.

[0041]

[0042] Formula IV Formula V

[0043] In compounds of formula IV and / or V, the wavy bonds represent unspecified configurations at the carbon atoms they are attached to.

[0044] In a more specific embodiment of the invention, the compounds of formulas IV and V, if present, are present at a level of less than 0.5 wt% of the mixture, and more specifically at a level of about 0.3 wt% or less.

[0045] In a specific embodiment of the present invention, a flavoring mixture is provided comprising 50-70% by weight of a compound according to formula I, 28-38% by weight of a compound according to formula II, and 0.5 wt% or less of a compound according to formula IV and 0.1 wt% or less of a compound according to formula V.

[0046] In a specific embodiment of the present invention, a flavoring mixture is provided comprising 50-70% by weight of a compound according to formula I, 28-38% by weight of a compound according to formula II, 0.1 wt% or less of a compound according to formula IIIa, 0.1 wt% or less of a compound according to formula IIIb, 0.5 wt% or less of a compound according to formula IV, and 0.1 wt% or less of a compound according to formula V.

[0047] The applicant found that the compounds of formulas I and II, which are the driving factors of odor characteristics and intensity, are in the absolute (8'S) configuration, while their respective (8'R) absolute configurations have lower enantiomeric power.

[0048]

[0049] (8'S)-I / II

[0050] In the compound (8'S)-I / II, wavy bonds represent unspecified configurations at the carbon atoms they are attached to, and dashed lines together with solid lines represent a single bond and a double bond.

[0051] In particular, the compound of formula I with the absolute configuration (1'R, 2S, 8'S) was found to have an odor detection threshold 27,700 times lower, and was therefore significantly more effective than its enantiomer with the absolute configuration (1'S, 2R, 8'R).

[0052]

[0053] (1'R, 2S, 8'S)-I (1'S, 2R, 8'R)-I

[0054] The enantiomers of the compounds of Formula I are shown in their absolute configurations.

[0055] Therefore, in a particular embodiment of a fragrance mixture containing a compound of formula I, the enantiomer of configuration (1'R,2S,8'S) is enantiomer in excess, for example, about 20% ee or higher, for example, about 40% ee or higher, or about 60% ee or higher.

[0056] In a specific embodiment of the present invention, the compound of formula II comprises isomer pairs.

[0057]

[0058] rel -(1'S, 6'R, 8'S)-II rel -(1'S, 6'S, 8'S)-II

[0059] in rel-(1'S, 6'R, 8'S) isomers relative to rel The -(1'S, 6'S, 8'S) isomer is enriched. The isomer with hydrogen at the 6' position and the bridging bicyclic system (1', 8') pointing in the same direction is dominant. In the representation with bold or dashed lines but without wedge bonds, the relative configurations of the hydrogen at position 6' and the bridging bicyclic system (1', 8') are depicted. Such representation does not indicate absolute configurations.

[0060] In a more specific implementation, the compound of formula II rel -(1'S, 6'R, 8'S) isomers and rel The ratio of the -(1'S, 6'S, 8'S) isomers is at least 9:1.

[0061] The applicant found that, compared to flavor blends containing higher levels of the 8'R isomer, flavor blends of compounds containing the diastereomeric enrichment of Formula II and 8'S isomers of Formula I and II exhibited superior properties and strength, thus, compared to commercial Ambermax. TM Compared to other ingredients, these flavor blends represent more efficient carbon utilization and more sustainable materials.

[0062]

[0063] (8'S)-I (8'S)-II

[0064] Compounds of formulas I and II are shown in their absolute (8'S) configuration.

[0065] It should be noted that for compounds of formula (I) and (II), due to the nomenclature rules, the absolute configuration of C1 in formula I vs. formula II changes, while the arrangement of substituents remains unchanged.

[0066] In particular, compounds of formula II with absolute configurations (1'S, 2S, 6'R, 8'S) were found to have odor detection thresholds that were 13,400 times lower, and were therefore significantly more effective than their enantiomers with absolute configurations (1'R, 2R, 6'S, 8'R).

[0067]

[0068] (1'S, 2S, 6'R, 8'S)-II (1'R, 2R, 6'S, 8'R)-II

[0069] Therefore, in a particular embodiment of a fragrance mixture containing a compound of formula II, the enantiomer with the configuration (1'S 2S, 6'R, 8'S) is enantiomer in excess, for example, about 20% ee or higher, for example, about 40% ee or higher, or about 60% ee or higher.

[0070] The effect of compounds on overall odor impression was demonstrated by comparing odor detection thresholds. The compounds listed below were obtained by a combination of semi-preparative achiral and chiral HPLC, and the concentrations of the HPLC isolates were determined by chiral GC-FID with external calibration (see Example 2).

[0071] Determining the odor detection threshold requires samples with high analytical and olfactory purity. Therefore, special care is taken to prevent trace contamination by effective stereoisomers that may affect the perception of weaker stereoisomers, and the olfactory purity of each HPLC isolate is verified using chiral GC sniffing techniques. If the main peak in the HPLC isolate is odorless during the olfactory purity assessment, the concentration of the stock solution is used to determine the GC odor detection threshold (GTH), and this value is reported as greater than (GTH > x ng). For all compounds whose olfactory purity can be confirmed by chiral GC sniffing analysis and whose main components are the odor carriers of the sample, GTH determination is performed using a non-chiral gas chromatographic olfactory assay, as described by Flachsmann et al. (2024).

[0072] The structures (including relative stereochemistry) of each compound I and II were assigned based on NMR data. Each peak can be separated into major and minor enantiomers on a chiral phase. The assignment of absolute stereochemistry is based on the observation that isolongifene is enriched to varying degrees in the (-)-enantiomer, but never in the (+)-enantiomer. This relates to its preparation from naturally occurring (+)-longifene, which is converted to (-)-isolongifene via a series of cation rearrangements and subsequent steps (shown below). Figure 1Depending on the acid strength and conditions, after the first protonation, the intermediate cation "Intermediate-1" can be partially racemized via Wagner-Meerwein rearrangement to its own enantiomer "enantiomer of Intermediate-1," ultimately yielding (+)-isophyllocarpanone. This explains the different levels of racemization (and therefore different ee) of isophyllocarpanone, but the always-rich (-)-isophyllocarpanone-scaled mixtures (as shown below), because racemization of Intermediate-1, in the worst case, can produce racemic mixtures without reversing the absolute stereochemistry. Based on this fundamental principle, the major enantiomer of each peak is designated as the absolute stereochemistry derived from (-)-isophyllocarpanone, while the minor enantiomer is designated as the absolute stereochemistry derived from (+)-isophyllocarpanone.

[0073] Furthermore, the activation of the ambergris receptor OR7C1 by different ambermax isomers was investigated. Only two compounds were shown to significantly activate the receptor (see Example 3). Fragrance blends rich in these influential isomers are more potent and effective in delivering the desired odor. It is also more sustainable due to its higher odor / carbon ratio compared to fragrance blends containing large amounts of odorless or less influential isomers.

[0074] In a second aspect of the invention, a fragrance composition is provided comprising a fragrance mixture as described herein and at least one other fragrance ingredient.

[0075] The at least one other flavoring ingredient may be a flavoring raw material, or it may be a functional component used in flavorings, such as a solvent. The at least one other flavoring ingredient does not include the ingredient Ambermax. TM .

[0076] In a specific embodiment of the invention, the at least one other fragrance ingredient is a solvent for the fragrance mixture. The solvent can be selected from any solvent used in the fragrance industry. In particular, the solvent is selected from triethyl citrate and Dowanol Tpm glycol ether (tripropylene glycol methyl ether).

[0077] In a specific embodiment of the present invention, the at least one other flavoring ingredient is a flavoring raw material. As described above, the ingredient Ambermax TM Its characteristics include delivering a rich ambergris aroma with hints of woody cypress. It can be paired with Ambermax. TM Flavoring ingredients that combine and complement the aroma profile can be used in a similar manner with the flavor mixtures of the present invention. Such flavoring ingredients include, but are not limited to:

[0078] Ambergris materials, such as Ambrofix (3a,6,6,9a-tetramethyldodecanonaphtho[2,1-B]furan), Amberketal (3,8,8,11a-tetramethyldodecano-5h-3,5a-epoxy-naphtho[2,1-C]oxacycloheptatriene), Amber Xtreme (decahydro-2,2,6,6,7,8,8-heptamethyl-2h-indene[4,5-B]furan), Ambrocenide (4h-4a,9-methylenechamomilecyclo[5,6-D]-1,3-m-dioxacyclopentene, octahydro-2,2,5,8), and Nimberol (2,2,6-trimethyl-α-propyl-cyclohexanepropanol).

[0079] Woody materials, such as cashmeran (6,7-dihydro-1,1,2,3,3-pentamethyl-4(5h)-indanone), ambroxan (Iso E Super) (2-acetyl-1,2,3,4,5,6,7,8-octahydro-2,3,8,8-tetramethylnaphthalene), Georgywood (2-acetyl-1,2,3,4,5,6,7,8-octahydro-1,2,8,8-tetramethylnaphthalene and 2-acetyl-1,2,3,4,5,6,7,8-octahydro-2,3,8,8-tetramethylnaphthalene), synthetic oakmoss (Evernyl) (methyl 2,4-dihydroxy-3,6-dimethylbenzoate), patchouli oil, cypress methyl ether, and Radjanol. (2-Ethyl-4-(2,2,3-trimethyl-3-cyclopenten-1-yl)-2-buten-1-ol), Javanol ((1-methyl-2-(1,2,2-trimethylbicyclo[3.1.0]hex-3-ylmethyl)cyclopropyl)methanol, Polysantol (trans-3,3-dimethyl-5-(2,2,3-trimethylcyclopenten-1-yl)penten-4-en-2-ol).

[0080] Musk materials, such as Galaxolide (1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylindo(5,6-c)pyran), ethylene brassinate (1,4-dioxane-5,17-dione), Habanolide (oxane-hexadecene-2-one), Muscenone (3-methylcyclopentadecanone), Velvione (5-cyclohexadecene-1-one), Sylkolide (2-(3,5-dimethylhex-3-en-2-yloxy)-2-methylpropyl cyclopropanecarboxylate), Ambrettolide (oxane-hexadecene-10-en-2-one), and Velvione. (5-Cyclohexadecene-1-one), Fixolide (6-acetyl-1,1,2,4,4,7-hexamethyl-1,2,3,4-tetrahydronaphthalene).

[0081] Powdered fragrance materials, such as coumarin (2h-1-benzopyran-2-one), vanillin (4-hydroxy-3-methoxybenzaldehyde), ethyl vanillin (3-ethoxy-4-hydroxybenzaldehyde), and ethyl maltol (3-hydroxy-2-ethyl-4H-pyran-4-one).

[0082] Other materials include methyl hedione (3-oxo-2-pentylcyclopentane acetate), dihydromyrcenol (2,6-dimethyl-7-octen-2-ol), linalool (3,7-dimethyl-1,6-octadien-3-yl acetate), Agrumex (2-tert-butylcyclohexyl acetate), gapponone (1-(5,5(3,3)-dimethylcyclohex-1-en-1-yl)pent-4-en-1-one), allyl amyl glycolate (2(3)-methylbutoxyacetic acid 2-propenyl ester), citronellol (3,7-dimethyl-6-octen-1-ol), geranium oil, and pomarose (5,6,7-trimethyloct-2,5-dien-4-one).

[0083] Furthermore, the flavoring ingredients may be selected from other auxiliaries commonly used in the art, such as diluents and preservatives. Such descriptions of flavoring ingredients are found in standard flavoring textbooks and references, such as "Perfume and Flavour Chemicals," S. Arctander, Allured Publishing Corporation, 1994, IL, USA, and later editions, which are incorporated herein by reference.

[0084] The adjuvants or excipients that can be used in the fragrance compositions of the present invention may refer to any of those conventional ingredients used in fragrance compositions for reasons other than their odor characteristics or not particularly related to their odor characteristics. For example, an adjuvant or excipient may be an ingredient that acts as an aid in processing one or more fragrance ingredients or a fragrance composition containing said ingredients, or it may improve the handling or storage of fragrance ingredients or fragrance compositions containing them. It may also be an ingredient that provides additional benefits such as imparting color or texture. It may also be an ingredient that imparts lightfastness or chemical stability to one or more ingredients contained in fragrance ingredients or fragrance compositions containing them. A detailed description of the properties and types of these substances that can be used in fragrance compositions containing these substances is not exhaustive, but it must be mentioned that the ingredients are well known to those skilled in the art. Examples include solvents and cosolvents; surfactants and emulsifiers; viscosity and rheology modifiers; thickeners and gelling agents; preservatives; pigments, dyes and colorants; extenders, fillers and reinforcing agents; stabilizers, fillers, acidifiers, buffers and antioxidants that resist the harmful effects of heat and light.

[0085] Any one or more flavoring ingredients or adjuvants or excipients used in the flavoring compositions according to the invention can be formulated in a delivery solvent to provide specific flavor or functional effects, such as altering the airy odor characteristics of the flavoring composition, if desired. The delivery solvent may include microcapsules or other solid supports to which one or more flavoring ingredients or adjuvants may be chemically or physically bound, respectively. Furthermore, one or more flavoring ingredients or adjuvants may be dissolved or dispersed in a matrix material for controlling the rate of release of the one or more ingredients therefrom. In another alternative embodiment, one or more ingredients or adjuvants may be loaded onto a porous matrix, such as cyclodextrin or zeolite or other inorganic materials. In yet another embodiment, one or more flavoring ingredients may be provided in the form of so-called pre-flavorings or flavor precursors, which may be activated under certain conditions such as moisture, heat, or light to release the flavoring ingredients in a controlled manner. In consideration of the foregoing, it should be understood that the flavoring composition may be at least partially in solid, gel, foam, and / or liquid form. If it is present in solid form, it may be in the form of granules, powder, or tablets.

[0086] The fragrance compositions according to the present invention are not necessarily limited to the above-mentioned fragrance ingredients. The fragrance compositions may contain one or more additional fragrance ingredients that are particularly well-integrated with the fragrance mixture. These fragrance ingredients may include any fragrance oils, alcohols, aldehydes, ketones, ethers, acetals, esters, lactones, macrocyclic compounds, and heterocyclic compounds commonly used in the fragrance industry, as well as any excipients or auxiliaries commonly used in combination with these fragrance ingredients, such as solvents.

[0087] The fragrance mixtures and fragrance compositions containing them can be used in the fragrance industry, particularly for imparting, enhancing or improving pleasant odor impressions, or for masking, reducing or eliminating unpleasant odor impressions, and such uses form another aspect of the invention.

[0088] Unpleasant odors that need to be reduced or eliminated may emanate from consumer products incorporating fragrance blends or compositions, or from the site where the consumer product is intended to be applied to treat odors, such as a human or animal body, or an inanimate surface.

[0089] The fragrance mixtures and compositions according to the invention can be used in fragrance compositions of the invention in a wide range of amounts, depending on the specific olfactory effect desired by a skilled fragrance maker. Due to its high impact, it can be used in small amounts to achieve an olfactory effect economically. Alternatively, due to its fresh, diffuse, transparent, and easily harmonized sensory aesthetic characteristics, it can be used in relatively high amounts as a harmonizer in various olfactory directions. Typically, the fragrance mixture can be used in a wide range of amounts, from 0.0001 to 90 wt%, preferably 0.001 to 50 wt%, more preferably 0.01 to 20 wt%, and most preferably 0.1 to 10 wt% of the fragrance composition.

[0090] A third aspect of the invention relates to consumer products containing a fragrance mixture or fragrance composition according to the first or second aspect of the invention.

[0091] The fragrance blend may be present in the consumer product in an amount of about 0.0001 wt% to about 30 wt% based on the total weight of the consumer product. However, these values ​​are given by way of example only, as experienced perfumers can also achieve the desired effect or create new fragrance blends with lower or higher concentrations.

[0092] The consumer products may be personal care products, such as fragrance extracts, perfumes, eau de toilette, aftershave, cologne, pre-shave products, splash cologne, scented fresh wipes, body care products, soaps, liquid shower gels, hair care products (e.g., shampoos, conditioners), deodorants, antiperspirants, hand creams and lotions, foot creams and lotions, hair removal creams and lotions, aftershave creams and lotions, tanning creams and lotions, and decorative cosmetics (e.g., makeup).

[0093] Consumer products can be cleaning products, such as acidic, alkaline and neutral detergents, fabric fresheners, ironing aids, liquid detergents, fabric softeners, laundry soaps, laundry sheets, disinfectants (such as surface disinfectants), air fresheners, aerosol sprays, waxes and polishes.

[0094] Consumer products can be household items, such as candles, lamp oil, incense, insecticides, repellents, and propellants.

[0095] Consumer products can be household care products, such as textile treatment products, ironing aids, cleaning cloths, laundry detergents, cleaning products, especially cleaning products for hard and / or soft surfaces, household cleaners, care products, laundry care products, fabric care products (e.g., selected from detergents, fabric conditioners, fragrance enhancers, drying sheets and fabric rinsing products for treating and scenting fabrics), room fragrances and air fresheners, conditioners, colorants, fabric conditioners, conditioning bases, pharmaceuticals, crop protection products, polishes, food, cosmetics, fertilizers, building materials, adhesives, bleaching agents, decalcifying agents, floor care products, cookware care products, leather care products or furniture care products, detergents, disinfectants, fragrances, release agents and / or precursors of the above products.

[0096] Consumer products can be cleaning products, for example:

[0097] Toilet cleaners or washroom cleaners, in other words, products used to clean washroom dishes and urinals, are preferably supplied in powder, block, tablet, or liquid form, and preferably gel. Among other typical ingredients such as surfactants, they typically include organic acids (e.g., citric acid and / or lactic acid) or sodium bisulfate, amide sulfuric acid, or phosphoric acid for removing limescale or urine stains.

[0098] Pipe cleaning products or drain cleaners. These are typically strongly alkaline products, usually used to remove pipe blockages containing organic matter such as hair, grease, food residue, soap deposits, etc. Adding Al or Zn powder can help generate effervescent H2 gas. Possible ingredients are typically alkalis, alkaline salts, oxidizing agents, and neutral salts. Powdered forms preferably also include sodium nitrate and sodium chloride. Liquid pipe cleaning products may also preferably include hypochlorite. Enzyme-based drain cleaners are also available. Acidic products are also possible.

[0099] Multi-purpose or general-purpose cleaners. These cleaners can be used universally on all hard surfaces in homes and businesses, and can be wet-cleaned or rubbed. Generally, they are neutral, slightly alkaline, or slightly acidic products, especially liquid products. Multi-purpose or general-purpose cleaners typically contain surfactants, builders, solvents and water-soluble growth promoters, dyes, preservatives, etc.; multi-purpose cleaners with special disinfecting properties. They also include active antimicrobial ingredients (e.g., aldehydes, alcohols, quaternary ammonium compounds, amphoteric surfactants, triclosan, etc.).

[0100] Hygiene cleaning agents. These products are used for cleaning bathrooms and toilets. Alkaline hygiene cleaning agents are preferred for removing greasy dirt, while acidic hygiene cleaning agents are particularly good for removing limescale. Hygiene cleaning agents also advantageously have considerable disinfecting properties, especially strongly alkaline hygiene cleaning agents containing chlorine;

[0101] Oven cleaners or grill cleaners are supplied in gel or foam spray form. They are typically used to remove burnt or charred food residue. Oven cleaners are preferably strongly alkaline formulations, using, for example, sodium hydroxide, sodium metasilicate, or 2-aminoethanol. Furthermore, they typically contain anionic and / or nonionic surfactants, water-soluble solvents, and in some cases, thickeners such as polycarboxylate and carboxymethyl cellulose.

[0102] Glass cleaners and window cleaners. These products are preferably used to remove dirt, especially oil stains, from glass surfaces. Preferably, they contain compounds such as anionic and / or nonionic surfactants (particularly, up to 5% by weight), ammonia and / or ethanolamine (particularly, up to 1% by weight), ethanol and / or 2-propanol, glycol ethers (particularly, 10-30% by weight), water, preservatives, dyes, antifogging agents, etc.

[0103] Consumer products can be cosmetics, such as (a) beauty and skin care products, especially bath products, skin cleansing and washing products, skin care products, eye cosmetics, lip care products, nail care products, intimate care products, and foot care products; or (b) cosmetics with specific effects, especially sunscreens, tanning products, bleaching products, deodorants, antiperspirants, hair removal products, shaving products, and fragrances; (c) cosmetic dental care products, especially dental and oral care products, dental care products, denture cleaners, and denture adhesives; or (d) cosmetic hair care products, especially shampoos, hair care products, styling products, and hair dyes.

[0104] A fourth aspect of the invention relates to a method for preparing a fragrance mixture according to a first aspect of the invention.

[0105] In U.S. Patent 7,550,417, in the ingredient Ambermax TM The starting material for its preparation is isophyllocarbazone. Isophyllocarbazone can be obtained as a mixture of two enantiomers, namely (-)-isophyllocarbazone and (+)-isophyllocarbazone.

[0106] The applicant has now surprisingly discovered that fragrance blends prepared from enantiomers enriched with isolonga ketones contain a higher proportion of the desired isomers and a relatively small amount of isomers that exhibit weak or undesirable odors, and therefore, such blends are more carbon efficient and thus sustainable.

[0107] In a more specific embodiment, (-)-isolonga ketone is in the form of an enantiomer excess of about 20% or more, such as 40% or more, such as 60% or more, in the form of a mixture of isomers.

[0108] In one specific embodiment of the present invention, the method for preparing the fragrance mixture includes the following steps:

[0109] - Provides isolongifanone in rich form of its (-)-isolongifanone isomer; for example, at least about 20% ee or higher, for example, about 40% ee or higher, or about 60% ee or higher enantiomeric excess;

[0110] - Using a base to diastereomeric (-)-isolonga ketone at the 6'-position to provide a balanced mixture of 6'R and 6'S diastereomers, wherein the 6'R diastereomer is the predominant diastereomer;

[0111] - The mixture of (-)-isolongyl ketone diastereomers was subjected to Wittig olefination to form ethylidene tetramethyltricycloundecane (ETTU) according to formula VII, accompanied by stereochemical inversion at the 6-position, such that the 6'RETTU diastereomer is the major diastereomer.

[0112] - The ETTU diastereomer mixture is reacted with formaldehyde or paraformaldehyde in a Prins reaction to provide a fragrance mixture comprising compounds of formula I and formula II.

[0113]

[0114] (-)-Isophyllocodanone(6'R)-VII

[0115] Formula (VI)

[0116] Selecting the (-)-isolonga ketone enriched by the above enantiomers as a raw material is an important step in the method for preparing the fragrance mixture of the present invention.

[0117] Furthermore, another aspect of the invention is formed by using enantiomer-enriched (-)-isolongafolone as a raw material in the preparation of the fragrance mixture according to the first aspect of the invention.

[0118] Following a thorough study of the complex stereochemistry of the fragrance mixtures and their preparation methods, the applicant discovered that the hydrogen at the 6'-position of the (-)-isophyllocarpanone ring system can be epimerized to form diastereomer pairs of reactive intermediates (a mixture of 6'S and 6'R (-)-isophyllocarpanones). The balanced distribution of the reactive intermediates makes the 6'R diastereomer the predominant diastereomer, which at first glance seems problematic, given that the 6'S diastereomer is advantageous in terms of its olfactory properties. However, the subsequent Wittig olefination reaction surprisingly reversed the stereochemistry at the 6'-position of the ring system.

[0119] Not wishing to be bound by any particular theory, the applicant believes that the 6'-diastereoisomer ratio of the (-)-isophyllocarne starting material has no effect on the 6'-diastereochemistry of Ambermax. The 6'-stereocenter of (-)-isophyllocarne undergoes epimerism at the Wittig stage, resulting in a significantly faster reaction of the 6'S diastereomer compared to the 6'R diastereomer. The result is C6' configuration inversion, which alters the distribution of ETTU diastereomers, making the 6'R diastereomer the overwhelmingly dominant diastereomer.

[0120] Therefore, despite the foregoing, the choice of enantiomer-enriched (-)-isolonga ketone as a starting material was unexpectedly advantageous, resulting in much higher levels of the desired isomer in the flavor blend than could have been achieved so far.

[0121] In one specific embodiment of the invention, the Vitish olefination can be carried out using triphenylphosphonium salts, such as fluorides, chlorides, bromides, iodides, acetates, perchlorates, tetrafluoroborates, hydroxides, phenolates, phosphates, carbonates, methanesulfonates, sulfonates, nitrates, and lithium, sodium, or potassium alkoxide bases, such as methanol, ethanol, isopropoxide, or tert-butoxide.

[0122] In one specific embodiment of the invention, the Vittigene oxidation can be carried out in an aromatic nonpolar solvent, such as benzene, toluene, ethylbenzene, xylene, naphthalene, and mixtures thereof. By using such a solvent, it is possible to significantly increase the total concentration of the reaction mixture. For example, by using xylene, the concentration of the reaction mixture can be increased by up to four times compared to cyclohexane. This allows for a solvent reduction of up to 75%, thereby reducing costs, waste, and overall productivity.

[0123] In a particular embodiment of the invention, the Vittigene oxidation can be carried out at elevated temperatures, i.e., above 30°C, for example in the range of 30°C to 120°C, for example in the range of 50°C to 100°C, for example at 80°C, depending on the solvent used. The increased temperature can lead to a significant reduction in reaction time compared to a reaction at room temperature, and thus to an increase in productivity.

[0124] In one particular implementation, the ETTU intermediate is obtained as a mixture of diastereomers in a ratio >9:1, wherein the 6'R diastereomer is the predominant diastereomer.

[0125] The ETTU intermediate is converted into a fragrance mixture according to the first aspect of the invention via the Prince reaction, wherein stereochemistry is retained at the 6'-position of the ring system.

[0126] In a specific embodiment of the present invention, the Prince reaction can be carried out using solid paraformaldehyde or an aqueous solution of formaldehyde.

[0127] In one specific embodiment of the invention, an aqueous formaldehyde solution is used in the method. This allows for easier handling on an industrial scale, as liquids are generally easier to load into reaction vessels. Furthermore, the use of aqueous formaldehyde results in cleaner reaction characteristics, particularly by reducing the content of the low-odor components represented by formulas III, IV, and V. Specifically, when aqueous formaldehyde is used, the levels of the products corresponding to formulas IV and V decrease from 2-3% to below 0.5%.

[0128] In one specific embodiment of the invention, solid paraformaldehyde is used in the method. Compared to aqueous formaldehyde, solid paraformaldehyde allows for higher reaction concentrations, resulting in lower production costs and increased productivity. Furthermore, the use of solid paraformaldehyde is associated with enhanced olfactory quality of the final product compared to the use of commercially available aqueous formaldehyde solutions. While the pure reaction with solid paraformaldehyde is known to produce higher levels of low-odor components corresponding to formulas III, IV, and V, it has been found that combining solid paraformaldehyde with a protonated additive effectively reduces the content of these components to below 1%. The protonated additive may include water, methanol, ethanol, isopropanol, butanol, or mixtures thereof.

[0129] Existing methods use water as a proton additive. However, the applicant has found that the reaction is preferably carried out in the presence of a proton additive consisting of a mixture of a polar protic organic solvent and water. More specifically, the polar protic organic solvent is a lower alcohol, and more particularly, methanol.

[0130] By using a mixture of polar protonic organic solvents and water as a proton additive, the formation of undesirable formate esters that adversely affect the olfactory quality of the desired product is reduced. Compared to using water as the sole proton additive, the formate content can be reduced by 10 times by using a mixture of water and methanol as the proton additive.

[0131] The invention will now be further described with reference to the following non-limiting embodiments. Variations and modifications that will be apparent to those skilled in the art are within the scope of the invention, which is defined by the appended claims.

[0132] Example 1: The effect of paraformaldehyde on product formation

[0133] Ambermax products contain the following compounds:

[0134] (1'R, 2S, 8'S)-I and (1'S, 2R, 8'R)-I:

[0135] 1 ¹H NMR (C6D6) δ: 3.43 (dd, J =10.0, 9.0 Hz, 1H), 3.34 (dd, J =10.0, 6.7Hz, 1H), 2.85-2.96 (m, 1H), 1.72-1.81 (m, 3H), 1.59-1.66 (m, 2H), 1.31-1.45(m, 3H), 1.17-1.22 (m, 1H), 1.19 (s, 3H), 1.18 (s, 3H), 1.13 (dd, J=9.5, 1.6Hz, 1H), 1.06-1.13 (m, 1H), 0.93 (s, 3H), 0.88 (d, J =6.7 Hz, 3H), 0.87 (s, 3H) ppm. 13 13C NMR (C6D6) δ: 149.2 (s), 125.3 (s), 65.7 (t), 57.4 (s), 49.8 (d), 43.0 (s), 38.0 (d), 36.4 (t), 34.1 (t), 31.3 (s), 29.5 (t), 27.8 (q), 26.5 (q), 26.3 (q), 25.3 (t), 24.5 (q), 20.8 (t), 15.7 (q) ppm.

[0136] (1’R, 2R, 8’S)-I and (1’S, 2S, 8’R)-I:

[0137] 1 1H NMR (C6D6) δ: 3.47 (dd, J =10.1, 8.2 Hz, 1H), 3.36 (dd, J =10.1, 7.3Hz, 1H), 2.93 (m, 1H), 1.81-1.89 (m, 1H), 1.73-1.79 (m, 1H), 1.67-1.73 (m, 1H), 1.57-1.65 (m, 2H), 1.31-1.44 (m, 3H), 1.28 (s, 3H), 1.17 (ddd, J =12.9, 6.3, 1.3 Hz, 1H), 1.11 (dd, J =9.8, 1.9 Hz, 1H), 1.08 (s, 3H), 1.01-1.07 (m, 1H), 0.92 (s, 3H), 0.88 (d, J =6.9 Hz, 3H), 0.84 (s, 3H) ppm. 13¹³C NMR (C₆D₆) δ: 149.7 (s), 125.1 (s), 66.0 (t), 57.4 (s), 49.6 (d), 43.0 (s), 38.0 (d), 36.4 (t), 34.0 (t), 31.1 (s), 29.7 (t), 27.4 (q), 27.0 (q), 26.6 (q), 25.4 (t), 24.3 (q), 20.9 (t), 15.3 (q) ppm.

[0138] (1’R, 2R, 6’S, 8’R)-II and (1’S, 2S, 6’R, 8’S)-II:

[0139] 1 ¹H NMR (C₆D₆) δ: 5.35 (dt, J J=5.7, 2.5 Hz, 1H), 3.34-3.39 (m, 1H), 3.21-3.27 (m, 1H), 2.39 (sxt, J J=6.5 Hz, 1H), 2.11 (m, 1H), 1.91 (br dd, J J=17.4, 4.7 Hz, 1H), 1.64-1.73 (m, 2H), 1.51 (ddd, J=17.4, 5.8, 2.0 Hz, 1H), 1.26-1.44 (m, 3H), 1.09-1.11 (m, 1H), 1.14-1.23 (m, 1H), 1.08 (d, J J=6.9 Hz, 3H), 1.08 (s, 3H), 1.00 (s, 3H), 0.89 (s, 3H), 0.83 (s, 3H) ppm. 13 ¹³C NMR (C₆D₆) δ: 140.8 (s), 122.0 (d), 69.1 (t), 55.7 (d), 55.3 (s), 51.4 (d), 39.6 (t), 39.0 (d), 37.9 (s), 36.2 (t), 33.8 (q), 32.7 (s), 27.0 (t), 24.9 (q), 24.2 (q), 23.9 (q), 22.8 (t), 16.0 (q) ppm.

[0140] No formation of compounds of formula IIIa and / or formula IIIb was observed. Therefore, based on the total weight of the flavor mixture, its content is less than 0.5 wt%, for example less than 0.2 wt%, and particularly less than 0.1 wt%.

[0141] The content of compounds IV and / or V depends on the formaldehyde used in the ETTU conversion step (Prince reaction).

[0142] a) Solid paraformaldehyde, solvent-free:

[0143] Solid paraformaldehyde (2.58 g, 128 mol%) was added to ETTU formula VII (16 g). The reaction was heated at 180 °C for 6 hours. The reaction mixture was cooled to room temperature to give crude Ambermax containing 2.8% formate ester impurities (compounds of formulas IV and V).

[0144] b) Solid paraformaldehyde, using water / MeOH as a proton additive:

[0145] Solid paraformaldehyde (2.58 g, 128 mol%) was added to ETTU formula VII (16 g). Water and methanol (1:2 by weight) were added to provide a formaldehyde / L solvent concentration of 366 g. The reaction was heated at 180 °C for 6 hours. The reaction mixture was concentrated under vacuum to remove water and methanol, giving crude Ambermax containing 0.2% formate ester impurities (compounds of formulas IV and V).

[0146] c) Water-containing formaldehyde

[0147] A 37 wt% (7.0 g, 128 mol%) aqueous formaldehyde solution was added to ETTU formula VII (16 g). The reaction was heated at 180 °C for 6 hours. The reaction mixture was concentrated under vacuum to remove water, giving crude Ambermax containing 1.1% formate ester impurities (compounds of formulas IV and V).

[0148] A comparison of the Prince reaction under three different conditions shows the effect of the protonated additive on the formation of compounds of formula IV and V. In Examples 1a, b, and c, the same amount of formaldehyde was added. The highest amount of compounds of formula IV and V was observed when solid paraformaldehyde was used without solvent (Example 1a). If the reaction was carried out using water as the protonated solvent (Example 1c), the formation of compounds of formula IV and V was reduced. If the reaction was carried out using a mixture of water and methanol as the protonated additive (Example 1b), the formation of these compounds could be further reduced.

[0149] Example 2: Isolation and characterization of Ambermax compounds:

[0150] A mixture of Ambermax compounds (10 mg / mL in acetonitrile, 4 μL injection volume) was pre-fractionated by achiral reversed-phase HPLC using a Dionex Ultimate 3000 equipped with a UV detector at 200 nm and a fraction collector AFC-3000. Separation was performed on an Ascentis® Express C18 column (Merck P / N 53829-U, 150 mm length, 4.6 mm inner diameter, 2.7 μm particle size, and 90 Å pore size) using isocratic elution with a mobile phase of water and acetonitrile (25:75, v / v) at a flow rate of 1 mL / min and a column temperature of 25 °C. The retention times of the fractions with peaks of interest were 10.70 min for fraction F2, 11.31 min for fraction F4, and 12.17 min for fraction F6 (see [link to relevant documentation]). Figure 2 ).

[0151] Analytical purity checks of the achiral reversed-phase HPLC fractions F2, F4, and F6 were performed on a Thermo Scientific TRACE1300 series gas chromatograph and a TriPlus 100 LS autosampler equipped with a cold on-column injector, flame ionization detector (FID), and achiral capillary columns VF-WAXms (Agilent P / N CP9210, polyethylene glycol stationary phase, 30 m long, 0.32 mm inner diameter, 0.5 μm film thickness). Hydrogen was used as the carrier gas at a constant pressure of 60 kPa. The column oven temperature program was set as follows: initial temperature of 35 °C for 2 min, followed by ramping to 250 °C at 3 °C / min, and holding at the final temperature for 10 min. The FID temperature was set at 270 °C, and the gas flow rates were 350 mL / min for air, 35 mL / min for hydrogen, and 40 mL / min for supplemental nitrogen.

[0152] GC-FID chromatograms of Ambermax (top) and achiral reversed-phase HPLC fractions F2 (second from top), F4 (third from top), and F6 (see) Figure 3 Fraction F2 contains peak P2 as the major component (tR 56.10 min) and peak P3 as the minor component (tR 56.88 min), fraction F4 contains peak P1.1 as the major component (tR 55.33 min) and peak P2 as the minor component (tR 56.09 min), and fraction F6 contains peak P1 as the major component (tR 55.29 min).

[0153] To obtain sufficient quantities of purified diastereomers P1, P1.1, and P2, 143 injections were performed to collect the achiral reversed-phase HPLC fractions F2 and F6, and 194 injections were performed to collect the achiral reversed-phase HPLC fraction F4. Subsequently, the combined collections of fractions F2, F4, and F6 were concentrated to dryness in a rotary evaporator, dissolved in hexane, and further purified by a chiral HPLC system equipped with a Dionex Ultimate 3000, featuring a UV detector at 200 nm, a Jasco OR-4090 optical rotation detector (ORD), and an AFC-3000 fraction collector. Separation was performed by injecting 2 μl of the hexane solution (concentrations between 1.3 and 10 mg / mL) onto a Regis (R,R) WHELK-O® 1 column (BGB Analytik P / N 1-780222-300, 150 mm length, 4.6 mm inner diameter, 3.5 μm particle size, and 100 Å pore size), using isocratic elution at a flow rate of 1 mL / min and a column temperature of 15 °C using a mobile phase of hexane and isopropanol (95:5, v / v). The combined fractions of the Ambermax stereoisomers with specified run numbers were concentrated to dryness in a rotary evaporator and dissolved in methyl tert-butyl ether (MtBE). The concentration of the MtBE stock solution was determined by chiral GC-FID with external calibration. The chiral GC-FID method is described in the following sections.

[0154] exist Figure 4 The image depicts chiral HPLC chromatograms of fractions F2 (top) with enantiomer pairs (-)-P2 and (+)-P2, fraction F4 (middle) with enantiomer pairs (-)-P1.1 and (+)-P1.1, and fraction F6 (bottom) with enantiomer pairs (+)-P1 and (-)-P1, exhibiting the optical rotation of the Ambermax enantiomers. The upper chromatogram is an ORD chromatogram, and the lower chromatogram is a UV detection at 200 nm. A slight retention time shift was observed when the two detectors were sequentially connected.

[0155] For all Ambermax stereoisomers, the olfactory purity of MtBE stock solutions was evaluated using chiral GC sniffing techniques. These chiral GC sniffing analyses were performed on a Thermo Scientific TRACE 1300 Series gas chromatograph and a TriPlus 100 LS autosampler equipped with a split injector (Givaudan in-house product) using a flame ionization detector (FID) and a sniffing port. The column effluent was split 1:1 between the FID and the sniffing port using an inert glass Y-split and an equal-length deactivated fused silica capillary (0.5 m long, 0.25 mm inner diameter, BGB Analytik). The transfer line to the sniffing port was heated to 250°C throughout the GC run. Separation was performed on a Hydrodex beta-3P column (Macherey-Nagel P / N 723358.25, 25 m long, 0.25 mm inner diameter). Hydrogen was used as the carrier gas at a constant flow rate of 1.5 ml / min. The inlet temperature was set at 230 °C, and the split ratio was 20:1. The column oven temperature program was set as follows: initial temperature of 50 °C held for 2 min, followed by a ramp to 230 °C at 2 °C / min, and held at the final temperature for 10 min. The FID temperature was set at 250 °C, and the gas flow rates were 350 ml / min for air, 35 ml / min for hydrogen, and 40 ml / min for supplemental nitrogen.

[0156] If the main peak of the MtBE stock solution is odorless during the olfactory purity assessment, the concentration of the stock solution is used to determine the GC odor detection threshold (GTH), and this value is reported as greater than (GTH>x ng).

[0157] For all Ambermax stereoisomers, the olfactory purity of the sample can be confirmed by chiral GC olfactory analysis, and the main component is the odor carrier of the sample. GTH determination is performed by achiral gas chromatography-olfactory assay, as described by Flachsmann et al. (2024), Ambrox through the Looking Glass: Chemoenzymatic Synthesis and GC-Olfactometric Analysis of 15 Ambrox Stereoisomers, Helvetica Chimica Acta, Vol. 107, No. 7.

[0158] Table 1 summarizes the olfactory purity test and GTH of the isolated Ambermax compounds.

[0159] Table 1:

[0160]

[0161] Example 3: Odor receptor activation by different Ambermax compounds

[0162] The human odor receptor OR7C1 is specifically activated by molecules with ambergris odor characteristics.

[0163] Therefore, the potency of the Ambermax isomer was determined by measuring the in vitro activation of the OR7C1 (S99G, V126I, S210P, V247L) monomeric variant, which is more sensitive to amber molecules than the standard OR7C1 variant reported in GenBank. Figure 5 As shown, compounds (1'R, 2S, 8'S)-I and (1'S, 2S, 6'R, 8'S)-II are the most effective compounds.

[0164] Compounds (1'R, 2S, 8'S)-I and (1'S, 2S, 6'R, 8'S)-II were found to be olfactorily pure, while other compounds contained trace amounts of the most potent compound. Therefore, the activation of OR7C1 (S99G, V126I, S210P, V247L) by other compounds is likely due to trace amounts of (1'R, 2S, 8'S)-I and (1'S, 2S, 6'R, 8'S)-II, rather than by these compounds themselves.

[0165] A DNA sequence encoding human OR7C1 (S99G, V126I, S210P, V247L) with an optimized C-terminal domain was synthesized by a DNA synthesis service provider (BioCat GmbH, Germany) (DNA sequence SEQ ID NO: 1 and amino acid sequence SEQ ID NO: 2) and inserted into pcDNA3.1(+) using BamHI and NotI restriction sites (Invitrogen, MA, USA), which are located downstream of the CMV promoter sequence (SEQ ID NO: 3) and before the bgh terminator sequence (SEQ ID NO: 4). This synthesized sequence or nucleotide sequence further contains a Kozak sequence (GCCACC) and a nucleotide sequence encoding a signal peptide (mmLucy-FLAG-rho) at its 5' end (SEQ ID NO: 5 and 6). The plasmid thus contains constitutively expressed OR7C1 genes.

[0166] OR7C1 (S99G, V126I, S210P, V247L) gene expression was performed in HEK293T cells stably transfected with DNA sequences encoding functional variants of human RTP1S (V227I) (SEQ ID NO: 7 and 8) and RTP2 (L220R) (SEQ ID NO: 9 and 10). These cells were seeded at a density of 10,000 cells / well in polyethyleneimine-coated 96-well plates (100 μl / well) and grown at 37°C in the presence of 5% CO2 for 24 h. The growth medium was DMEM (Gibco™, ThermoFisher Scientific, MA, USA) containing 9% fetal bovine serum (FBS) and penicillin / streptomycin.

[0167] 0.6 μg of OR7C1 (S99G, V126I, S210P, V247L) expression plasmid, 1 μg of empty pcDNA3.1(+) vector, and 1 μg of pGL4.29 (Promega) carrying the CRE-inducible luciferase gene were pipetted into microtubes. The DNA mixture was then diluted in 0.25 mL of OptiMEM medium (Gibco™, ThermoFisher Scientific, MA, USA) containing 5 μL of P3000 reagent. In parallel, 5 μL of Lipofectamine 3000 (Invitrogen) was diluted in 0.25 mL of OptiMEM medium. After pre-incubation for 5 min, the two mixtures were combined to prepare the transfection mixture, which was then incubated for another 25 min.

[0168] Replace 50 μl of growth medium with fresh DMEM containing 9% fetal bovine serum (FBS) and penicillin / streptomycin. Dilute the transfection mixture in 5 ml of OptiMEM medium and add 50 μl of the diluted mixture to each well (final total volume 150 μl). Incubate the cells at 37°C in the presence of 5% CO2 for a further 24 h to allow DNA uptake and expression of OR7C1 (S99G, V126I, S210P, V247L).

[0169] The activation of OR7C1 cells (S99G, V126I, S210P, V247L) by the test compounds was measured by removing 100 μl of growth medium and adding 50 μl of DMEM containing 9% FBS, penicillin / streptomycin, 2% DMSO, and different concentrations of the test substances. After incubation for 4.5 h, cells were lysed using 20 μl of passive lysis buffer (Promega), and luciferase signaling based on OR-dependent cAMP production was measured.

[0170] sequence:

[0171] The optimized C-terminal DNA sequence of OR7C1 (S99G, V126I, S210P, V247L) (SEQ ID NO 1):

[0172] ATGGAAACAGGAAATCAAACACATGCCCAAGAATTTCTCCTCCTGGGATTTTCAGCAACGTCAGAGATTCAGTTCATTCTCTTTGGGCTGTTCCTCTCCATGTACCTAGTCACTTTCACCGGGAACCTGCTCATCATCCTGGCCATATGCTCAGACTCCCACCTCCACACCCCCATGTACTTCTTCCTCTCCAACCTGTCTTTTGCTGACCTCTGTTTTACCTCCACGACTGTCCCAAAGATGTTACTGAATATACTGACACAGAACAAATTCATAACATATGCAGGCTGTCTCGGTCAGATTTTTTTTTTCACTTCATTTGGATGCCTGGACAATTTACTCTTGACCGTGATGGCCTATGACCGCTTCGTGGCCATCTGTCACCCCCTGCACTATACGGTCATCATGAACCCCCAGCTCTGTGGACTGCTGGTTCTGGGGTCCTGGTGCATCAGTGTCATGGGTTCCCTGCTCGAGACCTTGACTGTTTTGAGGCTGTCCTTCTGCACCGAAATGGAAATTCCACACTTTTTTTGTGATCTACTTGAAGTCCTGAAGCTCGCCTGTTCTGACACCTTCATTAATAACGTGGTGATATACTTTGCAACTGGCGTCCTGGGTGTGATTCCCTTCACTGGAATATTTTTCTCTTACTATAAAATTGTTTTCTCTATACTGAGGATTTCCTCAGCTGGGAGAAAGCACAAAGCGTTTTCCACCTGTGGTTCCCACCTCTCACTGGTCACCTTGTTCTATGGCACGGGCTTTGGGGTCTATCTCAGTTCTGCAGCCACACCATCTTCTAGGACAAGTCTGGTGGCCTCAGTGATGTACACCATGGTCACCCCCATGCTGAACCCCTTCATCTACAGCCTGAGGAACAAAGAAGTTAAAAAGGCCATAAAGAGGTTGTTCAAGAGAAAGTGCTGCAGGAGAAGGTGA

[0173] The optimized C-terminal amino acid sequence of OR7C1 (S99G, V126I, S210P, V247L) (SEQ ID NO 2):

[0174] METGNQTHAQEFLLLGFSATSEIQFILFGLLFLSMYLVTFTGNLLIILAICSDSHLHTPMYFFLSNLSFADLCFTSTTVPKMLLNILTQNKFITYAGCLGQIFFFTSFGCLDNLLLTVMAYDRFVAICHPLHYTVIMNPQLCGLLVLGSWCISVMGS LLETLTVLRLSFCTEMEIPHFFCDLLEVLKLACSDTFINNVVIYFATGVLGVIPFTGIFFSYYKIVFSILRISSAGRKHKAFSTCGSHLSLVTLFYGTGFGVYLSSAATPSSRTSLVASVMYTMVTPMLNPFIYSLRNKEVKKAIKRLFKRKCCRRR

[0175] CMV promoter DNA sequence (SEQ ID NO 3)

[0176] GTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTC

[0177] bgh terminator DNA sequence (SEQ ID NO 4)

[0178] CTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGG

[0179] mmLucy-FLAG-rho DNA sequence (SEQ ID NO 5)

[0180] ATGAGCCACCAGATCCTGCTGCTCCTGGCCCTGCTGACCCTAGGCCTGGCTGATTACAAGGACGACGACGATAAGATCGAATTGATGAACGGGACCGAGGGCCCAAACTTCTACGTGCCTTTCTCCAACAAGACGGGCGTGGTGGAATTC

[0181] Amino acid sequence of mmLucy-FLAG-rho (SEQ ID NO 6)

[0182] MSHQILLLLALLTLGLADYKDDDDKIELMNGTEGPNFYVPFSNKTGVVEF

[0183] DNA sequence of RTP1S(V227I) (SEQ ID NO 7)

[0184] ATGTGTAAAAGCGTGACCACAGATGAGTGGAAGAAAGTCTTCTATGAGAAGATGGAGGAGGCAAAGCCGGCTGACAGCTGGGACCTCATCATAGACCCCAACCTCAAGCACAATGTGCTGAGCCCTGGTTGGAAGCAGTACCTGGAATTGCATGCTTCAGGCAGGTTCCACTGCTCCTGGTGCTGGCACACCTGGCAGTCGCCCTACGTGGTCATCCTCTTCCACATGTTCCTGGACCGCGCCCAGCGGGCGGGCTCGGTGCGCATGCGCGTCTTCAAGCAGCTGTGCTATGAGTGCGGCACGGCGCGGCTGGACGAGTCCAGCATGCTGGAGGAGAACATCGAGGGCCTGGTGGACAACCTCATCACCAGCCTGCGCGAGCAGTGCTACGGCGAGCGTGGCGGCCAGTACCGCATCCACGTGGCCAGCCGCCAGGACAACCGGCGGCACCGCGGAGAGTTCTGCGAGGCCTGCCAGGAGGGCATCGTGCACTGGAAGCCCAGCGAGAAGCTGCTGGAGGAGGAGGCGACCACCTACACCTTCTCCCGGGCGCCCAGCCCCACCAAGTCGCAGGACCAGACGGGCTCAGGCTGGAACTTCTGCTCTATCCCCTGGTGCTTGTTTTGGGCCACGGTCCTGCTGCTGATCATCTACCTGCAGTTCTCTTTCCGTAGCTCCATCTAA

[0185] Amino acid sequence of RTP1S(V227I) (SEQ ID NO 8)

[0186] MCKSVTTDEWKKVFYEKMEEAKPADSWDLIIDPNLKHNVLSPGWKQYLELHASGRFHCSWCWHTWQSPYVVILFHMFLDRAQRAGSVRMRVFKQLCYECGTARLDESSMLEENIEGLVDNLITSLREQCYGERGGQYRIHVASRQDNRRHRGEFCEACQEGIVHWKPSEKLLEEEATTYTFSRAPSPTKSQDQTGSGWNFCSIPWCLFWATVLLLIIYLQFSFRSSI

[0187] RTP2 (L220R) DNA sequence (SEQ ID NO 9)

[0188] ATGTGTACCAGCTTGACCACTTGTGAGTGGAAGAAAGTCTTCTATGAGAAGATGGAGGTGGCAAAGCCAGCGGACAGCTGGGAGCTCATCATAGACCCCAACCTCAAGCCCAGTGAGCTGGCCCCTGGCTGGAAGCAGTACCTGGAGCAGCACGCCTCAGGCAGGTTCCACTGCTCCTGGTGCTGGCACACCTGGCAGTCTGCCCATGTGGTCATCCTCTTCCACATGTTCCTGGACCGCGCCCAGCGGGCGGGCTCGGTGCGCATGCGCGTCTTCAAGCAGCTGTGCTATGAGTGCGGCACGGCGCGGCTGGACGAGTCCAGCATGCTGGAGGAGAACATCGAGGGCCTGGTGGACAACCTCATCACCAGCCTGCGCGAGCAGTGCTACGAGGAGGATGGTGGCCAGTACCGCATCCACGTGGCCAGCCGCCCGGACAGCGGGCCGCATCGTGCAGAGTTCTGTGAGGCCTGCCAGGAGGGCATCGTTCACTGGAAGCCCAGCGAGAAGCTGCTGGAGGAGGAGGTGACCACCTACACCTCTGAAGCCTCCAAGCCGAGGGCCCAGGCGGGATCCGGCTACAACTTCTTGTCTCTTCGCTGGTGCCTCTTCTGGGCCTCTCTCTGCCTGCTCGTTGTTTACCTGCAGTTCTCCTTCCGCAGTCCTGCCTTCTTTTAG

[0189] RTP2 (L220R) Amino acid sequence (SEQ ID NO 10)

[0190] MCTSLTTCEWKKVFYEKMEVAKPADSWELIIDPNLKPSELAPGWKQYLEQHASGRFHCSWCWHTWQSAHVVILFHMFLDRAQRAGSVRMRVFKQLCYECGTARLDESSMLEE NIEGLVDNLITSLREQCYEEDGGQYRIHVASRPDSGPHRAEFCEACQEGIVHWKPSEKLLEEEVTTYTSEASKPRAQAGSGYNFLSLRWCLFWASLCLLVVYLQFSFRSPAFF

[0191] Example 4: X-ray data of (1'R, 2S, 8'S)-I and (1'S, 2S, 6'R, 8'S)-II

[0192] a) (1'R, 2S, 8'S)-I ((-)-P1)

[0193] A hexane solution of a compound of formula (1'R, 2S, 8'S)-I contained in a 1.5 mL vial was slowly evaporated with a low-nitrogen gas stream until dry. After the solvent had completely evaporated, needle-like crystals were then observed.

[0194] Table 2 shows the crystal data and structural refinement of the compound of formula (1'R, 2S, 8'S)-I.

[0195] Table 2:

[0196]

[0197] The title compound crystallizes in chiral space group I41. The absolute configuration was reliably determined by conventional least-squares refinement and Bayesian statistics using Bijvoet differences. Two independent molecules exist within the asymmetric unit. The two isomers exhibit the same configuration at the C atoms (labeled C2 / C8 / C13 and C20 / C26 / C31). S / R / S ()( Figure 6 ).

[0198] b) (1'S, 2S, 6'R, 8'S)-II ((+)-P2)

[0199] The compound of formula (1'S, 2S, 6'R, 8'S)-II was dissolved in a minimal amount of cyclohexane. It was then evaporated in a partially sealed screw-top vial. After complete evaporation of the solvent, crystals were observed.

[0200] Table 3 shows the crystal data and structural refinement of the compound of formula (1'S, 2S, 6'R, 8'S)-II.

[0201] Table 3:

[0202]

[0203] The title compound crystallizes in chiral space group P212121. The absolute configuration was reliably determined by conventional least-squares refinement and Bayesian statistics using Bijvoet differences. There are six independent molecules within the asymmetric unit. Figure 7a All molecules are labeled C2 / C8 / C9 / C13 (molecule 1, Figure 7b ), C20 / C26 / C27 / C31 (molecule 2, Figure 7c ), C38 / C44 / C45 / C49 (molecule 3, Figure 7d ), C56 / C62 / C63 / C67 (molecule 4, Figure 7e ), C74 / C80 / C81 / C85 (molecule 5, Figure 7f ) and C92 / C98 / C99 / C103 (molecule 6, Figure 7g The C atoms of ) exhibit the same relative configuration. S / S / R / S .

Claims

1. A spice mixture comprising compounds according to Formula I and / or Formula II Formula I Formula II In the compounds of formula I and / or II, the wavy bond represents an unspecified configuration at the attached carbon atom, and in the compounds of formula II, the 6'R diastereomer is enriched relative to its 6'S diastereomer.

2. The fragrance mixture according to claim 1, wherein the diastereomeric ratio of the 6'R diastereomeric component to the 6'S diastereomeric component in the compound of formula II is at least 9:

1.

3. The flavoring mixture according to any one of the preceding claims, wherein the enantiomeric excess of the compounds of Formula I and Formula II is about 20%ee or higher, for example about 40%ee or higher, or about 60%ee or higher.

4. A spice mixture comprising compounds according to Formula I and / or Formula II. Formula I Formula II The spice mixture contained no or substantially no compounds according to Formula III. Formula III, Furthermore, in the compounds of formulas I, II, and / or III, the wavy bonds represent unspecified configurations at the connected carbon atoms, and in the compound of formula III, the dashed lines are combined with the solid lines ( () represents a single bond and a double bond, and "substantially free of" means less than 0.5 wt% based on the total weight of the spice mixture.

5. A spice mixture comprising compounds according to Formula I and / or Formula II. Formula I Formula II It contains no or substantially no compounds according to formula IV or formula V. Formula IV Formula V In compounds of formulas IV and / or V, the wavy bonds represent unspecified configurations at the attached carbon atoms. And "substantially free of" means based on a total weight of less than 0.5 wt% of the spice mixture.

6. The flavor mixture according to any one of the preceding claims, comprising 50-70% by weight of a compound of formula I, 28-38% by weight of a compound of formula II, 0.1 wt% or less of a compound of formula IIIa, 0.1 wt% or less of a compound of formula IIIb, 0.5 wt% or less of a compound of formula IV and 0.1 wt% or less of a compound of formula V.

7. A spice composition comprising a spice mixture according to any one of the preceding claims and at least one additional spice ingredient.

8. A consumer product comprising a flavoring mixture according to any one of the preceding claims.

9. A method for preparing the spice mixture according to claim 1, comprising the following steps: - Provide an enantiomeric excess of at least about 20% ee or higher, such as about 40% ee or higher, or about 60% ee or higher (-)-isolongaenoketone (a compound of formula VI). (-)-Isophyllocarbazone, formula (VI); - Using a base to diastereomeric (-)-isolonga ketone at the 6'-position to provide a balanced mixture of 6'R and 6'S diastereomers, wherein the 6'R diastereomer is the predominant diastereomer; - The mixture of (-)-isolonga ketone diastereomers was subjected to Vittigene olefination to form ethylidene tetramethyltricycloundecane (ETTU) according to formula VII, accompanied by stereochemical inversion at the 6-position, such that the 6'R ETTU diastereomer is the major diastereomer. Equation VII; - The ETTU diastereomer mixture is subjected to a Prince reaction with formaldehyde or paraformaldehyde to provide a fragrance mixture comprising compounds of formula I and formula II.

10. The method of claim 9, wherein the Wittig reaction is carried out using triphenylphosphonium bromide or triphenylphosphonium chloride and sodium tert-butoxide or potassium tert-butoxide.

11. The method according to claim 9 or claim 10, wherein the reaction with formaldehyde or paraformaldehyde is carried out in the presence of a protonated additive, such as an alcohol, preferably methanol.

12. The method according to any one of claims 9 to 11, wherein the paraformaldehyde or formaldehyde is in excess relative to the proton additive / water mixture.

13. The method of claim 12, wherein the paraformaldehyde or formaldehyde is in excess by at least 1.2 relative to the proton additive / water mixture.

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