WO2012171175A1

WO2012171175A1 - Taste modifying compound

Info

Publication number: WO2012171175A1
Application number: PCT/CN2011/075732
Authority: WO
Inventors: Olivier Haefliger; Justin Jun XU; Yoyo Hui-Juan YUAN; Jian Zou
Original assignee: Firmenich Sa
Priority date: 2011-06-14
Filing date: 2011-06-14
Publication date: 2012-12-20

Abstract

The present invention relates to the use of Formula (I) where Y represents * denoting the point at which the moiety bonds to the bicyclic ring, in any one of its stereoisomenc forms or mixtures thereof to confer, enhance, impart and/or modify the tingling sensation.

Description

TASTE MODIFYING COMPOUND

Technical field

The present invention relates to the field of taste. More particularly, it concerns the use of a particular bicyclic compound as a taste-enhancing ingredient to impart or reinforce a tingling taste sensation.

The present invention also concerns compositions or articles containing at least one of the aforementioned compounds. Background and Prior art

Various extracts are known in the art for producing a tingling sensation. Typical plant extracts include extracts from garlic, radish, pepper, onion, horseradish, mustard, ginger and the like. Several compounds are also described as tingling, for instance spilanthol, sanshool, sanshoamide, chavicine, trans-pellitorin and piperine.

Tingling agents are also described in US 5545424 and US 6780443 (both to

Takasago International) and US 5407665 (the Procter and Gamble Company).

To the best of our knowledge, there is no disclosure in the prior art of the use of the bicyclic compound(s) of the present invention in order to provide a tingling sensation. Description of the invention

We have now surprisingly discovered that a compound having the formula

Formula (I)

in any one of its stereoisomeric forms or mixtures thereof imparts and/or reinforces a tingling taste sensation. For the avoidance of doubt, the asterisk denotes the point at which the moiety bonds to the ring.

Accordingly, the present invention provides the use of a compound of Formula (I) as defined above to confer, enhance, improve or modify the tingling sensation of a foodstuff.

According to another aspect, the present invention provides a method of imparting or reinforcing the tingling taste sensation of a flavoring composition or article comprising adding to the composition or article a compound according to Formula (I) as defined above.

Detailed Description of the Invention

The tingling compound according to Formula (I) may exist as a mixture of isomers thereof. Where the moiety, Y, is represented by the structure

it is preferably that there is an excess of the isomer represented by the structure according to Formula (II)

Formula (II)

In a highly preferred aspect, the compound according to Formula (I) comprises substantially only the compound according to Formula (II). This isomer is found to provide a particularly strong tingling sensation. The other isomer is also found to provide a tingling sensation but are typically much weaker.

Alternatively and/or additionally to the presence of a compound according to Formula (II), a compound according to Formula (III):

may also be present.

Optionally, the tingling compound according to the invention can be used in combination with one or more other tingling compounds. Examples of suitable additional tingling compounds include those selected from the group of spilanthol, sanshool, hydroxy a-sanshool, hydroxy β-sanshool, hydroxy γ-sanshool, sanshool I, sanshool II, sanshoamide, Japanese pepper extract, black pepper extract, chavicine, piperine, echinacea extract, northern prickly ash extract, Nepalese spice timur extract, red pepper oleoresin, 4-(l-menthoxymethyl)-2-phenyl-l,3-dioxolane and jambu oleoresin. Yet more tingling agents can be found in US 5545424.

In yet another aspect, the invention provides a flavored article comprising:

i) at least one compound of Formula (I), or a flavoring composition as defined above; and

ii) a foodstuff base.

In the context of the present invention, the term "tingling" denotes a spicy, slightly numbing, prickly effect upon consumption of a product.

The tingling sensation is perceived by stimulation of cell receptors involved in trigeminal perception and responsible for tingling perceptions. Measure of the tingling intensity can, for example, be performed using an assay containing cells that express tingling sensing receptors (TRPA1) and burning sensing receptors (TRPVl). Functional assays, measuring the increase of intracellular calcium levels can be performed to quantify receptors activity by the tingling compound.

Thus, the present invention concerns a method to confer, enhance, improve or modify the taste properties, as indicated above, of a flavoring composition or of a flavored article, which method comprises adding to said composition or article an effective amount of at least a compound of Formula (I). In the context of the present invention "use of a compound of Formula (I)" includes the use of any composition containing compound (I) and which can be advantageously employed in the flavor industry as active ingredient.

In another aspect, the invention provides a taste-modifying composition comprising: i) as taste-conferring or modifying ingredient, at least one compound according to Formula (I) above;

ii) at least one ingredient selected from the group consisting of a flavor carrier and a flavor base; and

iii) optionally at least one flavor adjuvant.

By "flavor carrier" we mean here a material which is substantially neutral from a flavor point of view, insofar as it does not significantly alter the organoleptic properties of flavoring ingredients. The carrier may be a liquid or a solid.

Suitable liquid carriers include, for instance, an emulsifying system, i.e. a solvent and a surfactant system, or a solvent commonly used in flavors. A detailed description of the nature and type of solvents commonly used in flavor cannot be exhaustive. Suitable solvents include, for instance, propylene glycol, triacetine, triethyl citrate, benzylic alcohol, ethanol, vegetable oils or terpenes.

Suitable solid carriers include, for instance, absorbing gums or polymers, or even encapsulating materials. Examples of such materials may comprise wall-forming and plasticizing materials, such as mono, di- or trisaccharides, natural or modified starches, hydrocolloids, cellulose derivatives, polyvinyl acetates, polyvinylalcohols, proteins or pectins, or yet the materials cited in reference texts such as H. Scherz, Hydrokolloids : Stabilisatoren, Dickungs- und Gehermittel in Lebensmittel, Band 2 der Schriftenreihe Lebensmittelchemie, Lebensmittelqualitat, Behr's VerlagGmbH & Co., Hamburg, 1996. Encapsulation is a well known process to a person skilled in the art, and may be performed, for instance, using techniques such as spray-drying, agglomeration, extrusion, coacervation and the like.

By "flavor base" we mean here a composition comprising at least one flavoring ingredient.

The nature and type of the flavoring ingredients present in the base do not warrant a more detailed description here, the skilled person being able to select them on the basis of his general knowledge and according to intended use or application and the desired organoleptic effect. In general terms, these flavoring ingredients belong to chemical classes as varied as alcohols, aldehydes, ketones, esters, ethers, acetates, nitriles, terpenoids, nitrogenous or sulphurous heterocyclic compounds and essential oils, and said perfuming co-ingredients can be of natural or synthetic origin. Many of these co- ingredients are in any case listed in reference texts such as the book by S. Arctander, Perfume and Flavor Chemicals, 1969, Montclair, New Jersey, USA, or its more recent versions, or in other works of a similar nature, as well as in the abundant patent literature in the field of flavor. It is also understood that said co-ingredients may also be compounds known to release in a controlled manner various types of flavoring compounds.

The composition may further comprise a flavor adjuvant. In the context of the present invention, the phrase "flavor adjuvant" denotes an ingredient capable of imparting additional added benefit that is not flavoring. Such additional benefits include, but are not limited to color, light resistance or stability, chemical stability, oxidation resistance and so on. A detailed description of the nature and type of adjuvant commonly used in flavoring bases cannot be exhaustive. Nevertheless, such adjuvants are well known to a person skilled in the art.

A composition consisting of at least one compound of Formula (I) and at least one flavor carrier represents an embodiment of the invention as well as a flavoring composition comprising at least one compound of Formula (I), at least one flavor carrier, at least one flavor base, and optionally at least one flavor adjuvant.

Moreover, a compound of Formula (I) can be advantageously incorporated into flavored articles to positively impart, or modify, the tingling taste of said articles. Thus, in yet another aspect, the present invention provides a flavored article comprising:

i) as taste-conferring or modifying ingredient, at least one compound of Formula (I), as defined above, optionally present as part of a flavoring composition; and

ii) a foodstuff.

In the context of the present invention "foodstuff denotes a product that is taken orally and may either be swallowed, such as a food or beverage, or spat out, such as a mouthwash or toothpaste. Therefore, a flavored article according to the invention comprises one or more compounds according to Formula (I), as well as optional benefit agents, corresponding to taste and flavor profile of the desired edible product, e.g. a chocolate or a toothpaste.

The nature and type of the constituents of the foodstuffs or beverages do not warrant a more detailed description here, the skilled person being able to select them on the basis of his general knowledge and according to the nature of said product. The proportions in which the compounds according to the invention can be incorporated into the various aforementioned articles or compositions vary within a wide range of values. These values are dependent on the nature of the article to be flavored and on the desired organoleptic effect as well as the nature of the co-ingredients in a given base when the compounds according to the invention are mixed with flavoring co- ingredients, solvents or additives commonly used in the art.

Thus, use levels of the composition according to the invention may range from about 0.5 to 500 ppm, more preferably from about 1 to 100 ppm and most preferably from 3 to 50 ppm, when incorporated into a food or beverage.

A particularly preferred product into which the composition according to the invention can be incorporated is a toothpaste since toothpastes are typically alkaline (p.H. in excess of 7) and the compound according to the invention is found to be even more stable at higher p.H. values, such as above 7.

Nevertheless, if it is desired to incorporate the compound according to Formula (I) into a foodstuff or beverage or other composition that has an acidic pH then it can be very advantageous to encapsulate the compound. This protects the compound from the acidic medium and provides longer-term stability.

Examples

The invention will now be described in further detail by way of the following example, wherein the abbreviations have the usual meaning in the art, the NMR spectral

13

data were recorded in CDCI3, with a 400MHz machine for C, the chemical displacements, δ, are indicated in ppm with respect to TMS as standard, and the coupling constants, J, are expressed in Hz.

Example 1

Extraction of a Compound according to the Invention Commercially available pomelos (Citrus maxima) were peeled, paying close attention to maximizing the amount of flavedo and minimizing the amount of albedo. 638.82 g of this starting material were mixed with 600 mL of deionized water and reduced to a paste using a food processor. The mixture was transferred to a large beaker, and 500 mL of ethanol were added. After thorough mixing, the suspension was filtered.

The supernatant obtained after the filtration was transferred to a round-bottom flask. The ethanol was removed by vacuum distillation, and the remaining aqueous phase was extracted 3 times with ethyl acetate.

The residue remaining after the filtration was extracted two times with 600 mL of ethyl acetate.

All ethyl acetate phases resulting from the liquid-liquid extraction and from the solvent extraction were pooled. They were dried and neutralized with an adequate amount of Na₂S0₄ and KHC0₃ and then evaporated to dryness.

The remaining residue was extracted with dichloromethane. The insoluble fraction, which contained large amounts of polyphenol derivatives, was discarded. The dichloromethane solution was dried and neutralized with an adequate amount of Na₂S0₄ and KHCO3 and then evaporated to dryness. 2.8 gram of extract was obtained.

The solid obtained after the extraction was dissolved in 14 mL of dichloromethane. The solution was passed through a small amount of Na₂S0₄. The resulting solution was subjected to preparative gel permeation chromatography. A 25 mm wide and 35 cm long column packed with 50 g of Bio-Beads® S-X3 size exclusion material was eluted at 5 mL/min with dichloromethane. 5 mL aliquots of the crude solution were injected at a time. The fractions eluting between 17 and 19 minutes during each injection were pooled, neutralized with a little Na₂S0₄, filtered, and evaporated to dryness. 358.6 mg of dried residue were obtained.

The 358.6 mg of residue obtained as described above were dissolved in 1.5 mL of THF. The mixture was further fractionated by semi-preparative HPLC using a Nova-Pak HR CI 8 (Waters, 6μηι, 6θΑ, 7.8 ^χ 300 mm ) column held at 30 °C and eluted at 4 mL/min with a gradient of water and acetonitrile.

Table 1

Time (min) % Acetonitrile

1 20

16 95

16.5 95

16.6 20

19 20 50 μΐ, aliquots were injected at a time, and a total of 40 injections were performed. The fractions eluting between 9.5 and 10 minutes during each run were pooled and dried under vacuum, affording 150 mg of crude product containing the compound according to the invention.

The crude product was further purified through a second semi-preparative high- performance liquid chromatography step. The same setup described above was used except that elution was performed isocratically with 55 % of water and 45 % of acetonitrile, with a run time of 10 minutes. The fractions eluting between 5.8 and 6.8 minutes during each run were pooled and freeze dried, affording 110 mg of highly pure compound according to Formula (I).

The product was analyzed as follows:

ESI-MS-MS analysis

5 μΐ. of a 20ppm of solution in ethanol of the purified meranzin solution were injected into an LC/ESI-MS-MS (hybrid triple quadrupole / ion trap) system. An Uptisphere 5-ODB column (150mm x 2.1mm) was used, which was eluted at 0.5 mL/min with a gradient of water + 0.1% FA and acetonitrile + 0.1% FA.

Table 2

Two mass spectrometric experiments were conducted in parallel, a MSI full scan over the m/z range from 50 to 500, and a product ion scan from the precursor ion at m z 261 over the m/z range from 70 to 270.

Positive ESI-MS: 261 [M+H]⁺(61), 278 [M+NH₄]⁺ (100), 283 [M+Na]⁺;

Positive ESI-MS-MS (261), collision energy 35 V: 243(100), 189 (81), 177 (8) UV/Vis spectroscopy

The UV/Vis spectrum of the purified compound was measured from a 12 mg/L solution in ethanol. A local absorbance maximum was observed at 322 nm (0.710 AU), corresponding to an extinction coefficient of 1.54xl0⁴L-mor¹-cm^"1.

NMR spectroscopy

The purified compound was characterized by H- and C-NMR spectroscopy.

Table 3

High resolution mass spectrometry

The molecular formula of the purified compound was confirmed by high resolution mass spectrometry. HR-ESI-MS (pos.): 261.11237 ([M+H]⁺, Ci₅H₁₇0₄ ⁺; calc. 261.11214; Error: -0.90(ppm))

The infrared spectrum of the purified compound was measured from a KBr pellet.

Absorption bands were observed among others at 3015, 2951, 1710, 1603, 1565, 1300, 1254, 1118, 836 cm-¹.

X-ray spectroscopy

The absolute configuration of the purified compound was elucidated by x-ray spectroscopy (Cu Ka). The S configuration of the compound according to Formula (II) was confirmed.

Example 2 Synthesis of enantiomeric comound according to Formula (II)

938.5 mg of osthol (CAS number 484-12-8) were weighed and placed into 250 mL three- necked round bottom flask. Then 40 mL of 10% NaHC0₃ were added into the flask and stirred well while the reaction mixture was cooled using an ice bath. 1392 mg of mCPBA (m-chloroperbenzoic acid) were dissolved in 20 mL of dichloromethane, and then added dropwise into the flask at last. The reaction was allowed to proceed during about 1.5 hours.

About 200 mL of saturated NaHC0₃ solution and about 200 mL of diethyl ether were added to the reaction system. The mixture was then transferred into a separating funnel. The organic layer was washed with saturated NaHC0₃ 3 times to remove the mCPBA, then washed with sodium sulfite solution to remove the peroxide, and finally washed 3 times with saturated NaCl solution until the pH was neutral. The organic layer was dried with anhydrous Na₂S0₄ , then after filter, concentrated on Rotavapor(600 mbar, 30°C). The solvent was then removed and 0.979 g of crude enantiomeric sample was obtained.

The crude enantiomeric compound was purified by flash chromatography using a silica column (Interchim PuriFlash™ 50μηι, 80g, 20 cmx 3cm ID). The crude sample was dissolved in 3 mL of EtOAc, loaded onto the column, and eluted at 20 mL/min with a mixture of 65% cyclohexane and 35% EtOAc. The fractions eluting between 22.5 and 31.5 min were pooled, treated with Na₂S0₄, filtered, and dried. 712 mg of pure enantiomeric mixture of the compound according to Formula (II) were obtained.

The enantiomers of the compound according to Formula (II) were separated from the crude enantiomeric mixture by preparative chiral liquid chromatography using a 250 x 20 mm i.d. Chiralcel OD (Daicel Chemical Industries) column eluted at 4 mL/min with 80% cyclohexane and 20% isopropanol. 20 aliquots of a 0.2 mg/n L solution of pure enantiomeric compound were injected. The fractions eluting between 29.4 and 31.0 min, which contained the R-enantiomer, were pooled from 12 injections. Finally, 9.8 mg of pure R-enantiomer were obtained.

Example 3

Sensory Evaluation of the Compound According to Formula (II) Sensory evaluation of the tingling effect of various test solutions was conducted with 25 panelists. The panelists were asked to keep the sample solutions in their mouth for 15 seconds, and then to spit them out without swallowing.

The panelists were asked to assess the following parameters:

Tstart: Time when the tingling sensation appears

Imax: Maximum perceived intensity of the tingling sensation rated on a scale of 0 (no tingling sensation) to 10 (strongest tingling sensation):

Tmax: Time at which the tingling sensation is maximal

Tdecrease: Time at which the tingling sensation decreases after the Imax

Drnin and/or Tend: Intensity perceived at 3 minutes (the end of the test) or time when the sensation disappears if before 3 minutes.

Qualitative comments: nature of off-notes, any remarks.

Three samples were compared:

Sample 1 : Blank solution (500 ppm food-grade ethanol) in drinking water.

Sample 2: 10 ppm of purified (S)-enantiomer of compound according to Formula (II) in drinking water, prepared from a stock solution in food-grade ethanol so as to have a final concentration of 500 ppm of food-grade ethanol.

Sample 3: 10 ppm of synthetic (R)-enantiomer (compound according to Formula (II) in drinking water, prepared from a stock solution in food-grade ethanol so as to have a final concentration of 500 ppm of food-grade ethanol.

Descriptive statistics showed the following results (average of the entire panel with confidence intervals at 5%).

Table 4

The differences between the three sample solutions on each parameter were then analyzed by Analysis of Variance with Duncan test at 5%.

Sample 1 was rated as a little tingling (Imax=2) and the sensation disappeared after about

2 min.

Sample 2 was perceived as powerfully tingling (Imax = 7). Significant differences were observed on Tmax, Imax, Tdecrease, Tend and lend between samples 2 and 1. The tingling effect was very long-lasting with an intensity of 4 by the end of the test after

3 minutes.

Sample 3 was perceived as significantly different from sample 1 in terms of Tstart, Tmax, Tdecrease and Tend. It also showed a slight increase in tingling sensation compared to sample 1.

Sample 2 was perceived as being significantly more tingling and having a longer lasting tingling sensation than sample 3. The perception of sample 2 compared to sample 3 was not significantly different on Tstart, Tmax and Tend.

The qualitative comments confirmed the stronger tingling effect of sample 2 compared to sample 3.

Claims

1. Use of a compound of Formula (I)

Formula (I)

where Y represents

or

in any one of its stereoisomeric forms or mixtures thereof to confer, enhance, impart and/or modify the tingling sensation.

2. Use according to claim 1, wherein the compound is a mixture of stereoisomers according to Formula (I) and there is an excess of the stereoisomer according to Formula (II)

Formula (II)

3. Use according to claim 1, wherein said compound is the stereoisomer according to Formula (II)

Formula (II)

4. Use according to any one of the preceding claims wherein the compound according to formula (I) comprises a compound according to Formula (III)

Formula (III)

5. Use according to any one of the preceding claims wherein the compound or mixture of compound is/are encapsulated.

6. Use according to any one of the preceding claims wherein the compound according to Formula (I) is present in an amount of from 0.5 to 500 ppm in a product into which it is incorporated.

7. A taste-modifying composition comprising:

i) as taste-modifying ingredient, at least one compound according to Formula (I) as defined in any one of the preceding claims;

iii) optionally at least one flavor adjuvant.