ORAL PRODUCT

FIELD

The present disclosure relates to an oral product, a process for producing the oral product, as well as to pouched products and packages comprising said oral product.

BACKGROUND

Tobacco may be enjoyed in a so-called “smokeless” form. Particularly popular smokeless tobacco products are employed by inserting some form of processed tobacco or tobacco-containing formulation into the mouth of the user. Conventional formats for such smokeless tobacco products include moist snuff, snus, and chewing tobacco, which are typically formed almost entirely of particulate, granular, or shredded tobacco, and which are either portioned by the user or presented to the user in individual portions, such as in single-use pouches or sachets. Other traditional forms of smokeless products include compressed or agglomerated forms, such as plugs, tablets, or pellets. Alternative product formats, such as tobacco-containing gums and mixtures of tobacco with other plant materials, are also known. See for example, the types of smokeless tobacco formulations, ingredients, and processing methodologies set forth in U.S. Pat. Nos. 1,376,586 to Schwartz; 4,513,756 to Pittman et al.; 4,528,993 to Sensabaugh, Jr. et al.; 4,624,269 to Story et al.; 4,991,599 to Tibbetts; 4,987,907 to Townsend; 5,092,352 to Sprinkle, III et al.; 5,387,416 to White et al.; 6,668,839 to Williams; 6,834,654 to Williams; 6,953,040 to Atchley et al.; 7,032,601 to Atchley et al.; and 7,694,686 to Atchley et al.; U.S. Pat. Pub. Nos. 2004/0020503 to Williams; 2005/0115580 to Quinter et al.; 2006/0191548 to Strickland et al.; 2007/0062549 to Holton, Jr. et al.; 2007/0186941 to Holton, Jr. et al.; 2007/0186942 to Strickland et al.; 2008/0029110 to Dube et al.; 2008/0029116 to Robinson et al.; 2008/0173317 to Robinson et al.; 2008/0209586 to Neilsen et al.; 2009/0065013 to Essen et al.; and 2010/0282267 to Atchley, as well as WO2004/095959 to Arnarp et al., each of which is incorporated herein by reference.

Smokeless tobacco product configurations that combine tobacco material with various binders and fillers have been proposed more recently, with example product formats including lozenges, pastilles, gels, extruded forms, and the like. See, for example, the types of products described in U.S. Pat. App. Pub. Nos. 2008/0196730 to Engstrom et al.; 2008/0305216 to Crawford et al.; 2009/0293889 to Kumar et al.; 2010/0291245 to Gao et al; 2011/0139164 to Mua et al.; 2012/0037175 to Cantrell et al.; 2012/0055494 to Hunt et al.; 2012/0138073 to Cantrell et al.; 2012/0138074 to Cantrell et al.; 2013/0074855 to Holton, Jr.; 2013/0074856 to Holton, Jr.; 2013/0152953 to Mua et al.; 2013/0274296 to Jackson et al.; 2015/0068545 to Moldoveanu et al.; 2015/0101627 to Marshall et al.; and 2015/0230515 to Lampe et al., each of which is incorporated herein by reference.

All-white snus portions are growing in popularity, and offer a discrete and aesthetically pleasing alternative to traditional snus. Such modern “white” pouched products may include a bleached tobacco or may be tobacco-free.

It would be desirable to provide an oral product configured for oral use which may deliver active ingredients and flavor to the consumer in an enjoyable form, such as in the form of a pouched product.

BRIEF SUMMARY

In accordance with some embodiments described herein, there is provided an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; wherein the oral product comprises no more than about 5% by weight of a tobacco material, excluding any nicotine present, based on the total weight of the product; and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes.

In accordance with some embodiments described herein, there is provided a pouched oral product comprising a saliva permeable pouch and an oral product as defined herein incorporated within the pouch.

In accordance with some embodiments described herein, there is provided a package containing an oral product as defined herein or at least one pouched oral product as defined herein.

In accordance with some embodiments described herein, there is provided a process for preparing an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; wherein the oral product comprises no more than about 5% by weight of a tobacco material, excluding any nicotine present, based on the total weight of the product; and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes, the process comprising:

a. providing at least one filler or bulking agent and at least one flavor glycoside, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; and
b. contacting the at least one filler or bulking agent and at least one flavor glycoside to provide the oral product.

In accordance with some embodiments described herein, there is provided a use of a flavor glycoside to prolong flavor of an oral product, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond, and the product comprises no more than about 5% by weight of a tobacco material, excluding any nicotine present, based on the total weight of the product.

In accordance with some embodiments described herein, there is provided a use of a flavor glycoside to change the flavor released from an oral product over a period of time, wherein the oral product comprises a flavor glycoside and one or more additional flavoring agents, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond and wherein the one or more additional flavoring agents is different from the flavoring agent in the flavor glycoside.

In accordance with some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; wherein the flavoring agent is selected from the group consisting of geraniol, citronellol, nerol, maltol, ethylmaltol, fenchol, homofuraneol, furaneol, norfuraneol, 1-octen-3-ol, borneol, linalool, farnesol, hydroxycitronellol, 3,7-dimethyloctanol, myrcenol, lavandulol, nerolidol, terpineol, alpha-terpineol, menthol, thymol, carvacrol, myrtenol, carveol, santalol, piperitol, perillyl alcohol, patchouli alcohol, hexanol, 3-cis-hexanol, cis-3-hexen-1-ol, phenylethanol, eugenol, sesamol, sotolone, maple furanone, methyl anthranilate, guaiacol, raspberry ketone, 2-methoxy-4-vinylphenol, 4-ethylguaiacol, benzylalcohol, homofuraneol, and combinations thereof, and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes.

In accordance with some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside; and (ii) at least one tobacco material; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; and wherein the at least one tobacco material is present in an amount of from about 0.001 % to about 4.5% by weight of the oral product.

In accordance with some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside; and (ii) at least one tobacco material, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond, and wherein the flavor glycoside is obtained from a biotechnological process.

These and other features, aspects, and advantages of the disclosure will be apparent from a reading of the following detailed description together with the accompanying drawings, which are briefly described below. The invention includes any combination of two, three, four, or more of the above-noted embodiments as well as combinations of any two, three, four, or more features or elements set forth in this disclosure, regardless of whether such features or elements are expressly combined in a specific embodiment description herein. This disclosure is intended to be read holistically such that any separable features or elements of the disclosed invention, in any of its various aspects and embodiments, should be viewed as intended to be combinable unless the context clearly dictates otherwise.

For ease of reference, these and further aspects of the present invention are now discussed under appropriate section headings. However, the teachings under each section are not necessarily limited to each particular section.

BRIEF DESCRIPTION OF THE DRAWINGS

Having thus described aspects of the disclosure in the foregoing general terms, reference will now be made to the accompanying drawings, which are not necessarily drawn to scale. The drawings are exemplary only, and should not be construed as limiting the disclosure. Embodiments of the invention will now be described, by way of example only, with reference to accompanying drawings, in which:

FIG. 1 shows the amino acid sequences of terpene glycosyl transferases VvGT14 and VvGT15 from Vitis vinifera. (A): Amino acid sequence of terpene glycosyl transferase VvGT14 (SEQ ID NO: 1). (B): Amino acid sequence of terpene glycosyl transferase VvGT15 (SEQ ID NO: 2).

FIG. 2 is a cross-sectional view of a pouched product embodiment, taken across the width of the product, showing an outer pouch filled with a composition of the present disclosure.

FIG. 3 shows a chromatogram of raspberry ketone glycoside at different incubation times.

FIG. 4 is a graph showing the amount of raspberry ketone remaining in a product containing raspberry ketone glycoside after different incubation times and when placed in the mouths of three different testers.

DETAILED DESCRIPTION

It is to be understood that this invention is not limited to the particular configurations, process steps, and materials disclosed herein as such configurations, process steps, and materials may vary somewhat. It is also to be understood that the terminology employed herein is used for the purpose of describing particular embodiments only and is not intended to be limiting since the scope of the present invention will be limited only by the appended claims and equivalents thereof.

As used in this specification and the claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Reference to “dry weight percent” or “dry weight basis” refers to weight on the basis of dry ingredients (i.e., all ingredients except water). Reference to “wet weight” refers to the weight of the composition including water. Unless otherwise indicated, reference to “weight percent” (or “% by weight”) of a composition reflects the total wet weight of the composition (i.e., including water).

In this specification, unless otherwise stated, the term “about” modifying the quantity of an ingredient in the tobacco material or tobacco compositions of the invention or employed in the methods of the invention refers to variation in the numerical quantity that can occur, for example, through typical measuring and liquid handling procedures used for making concentrates or use solutions in the real world; through inadvertent error in these procedures; through differences in the manufacture, source, or purity of the ingredients employed to make the tobacco material or tobacco compositions, or to carry out the methods; and the like. The term “about” also encompasses amounts that differ due to different equilibrium conditions for a composition resulting from a particular initial mixture. Whether or not modified by the term “about”, the claims include equivalents to the quantities.

Oral Product

As described herein, there is provided an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; wherein the oral product comprises no more than about 5% by weight of a tobacco material, excluding any nicotine present, based on the total weight of the product; and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes.

The oral product is configured for oral use, and thus for insertion into the user’s mouth (i.e. oral cavity). In this specification, unless otherwise stated, the term “oral” in connection to a product refers to a product which, in normal use, is suited to be placed somewhere in the oral cavity of the user, for example under the lips, in the same way as moist snuff products are generally used.

The oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes, such as at least about 10 minutes, such as at least about 15 minutes, such as at least about 20 minutes, such as at least about 30 minutes, such as at least about 45 minutes, such as at least about 1 hour. The oral product may be configured to be retained in the oral cavity of a user during use for a period of from about 5 minutes to 1 hour, such as from about 15 minutes to 45 minutes.

The ranges provided herein provide preferred amounts of each of the components. Each of these ranges may be taken alone or combined with one or more other component ranges to provide a preferred aspect of the invention.

Filler or Bulking Agent

As described herein, the oral product comprises at least one filler or bulking agent. The filler or bulking agent may be any suitable filler or bulking agent for preparing an oral product. Fillers, for example, may fulfil multiple functions, such as enhancing certain organoleptic properties such as texture and mouthfeel, enhancing cohesiveness or compressibility of the product, and the like.

In some embodiments, the at least one filler or bulking agent is or comprises a bulking agent. In some embodiments, the at least one filler or bulking agent is or comprises a filler.

In some embodiments, the filler or bulking agent is a porous particulate material and is cellulose-based. For example, the filler or bulking agent may be a non-tobacco plant material or derivative thereof, including cellulose materials derived from such sources. Examples of cellulosic non-tobacco plant material include cereal grains (e.g., maize, oat, barley, rye, buckwheat, and the like), sugar beet (e.g., FIBREX® brand filler available from International Fiber Corporation), bran fiber, and mixtures thereof.

In some embodiments, the oral product comprises at least one filler. In some embodiments, the filler is a cellulose material selected from the group consisting of maize fiber, oat fiber, barley fiber, rye fiber, buckwheat fiber, sugar beet fiber, bran fiber, bamboo fiber, wood pulp fiber, cotton fiber, citrus pulp fiber, grass fiber, willow fiber, poplar fiber, cocoa fiber, derivatives thereof, and combinations thereof. In some embodiments, the filler is a cellulose material selected from the group consisting of maize fiber, oat fiber, sugar beet fiber, bamboo fiber, wood pulp fiber, cotton fiber, grass fiber, derivatives thereof, and combinations thereof. In some embodiments, the filler is a cellulose material selected from the group consisting of sugar beet fiber, wood pulp fiber, bamboo fiber, derivatives thereof, and combinations thereof.

In some embodiments, the filler is derived from any of maize fiber, oat fiber, barley fiber, rye fiber, buckwheat fiber, sugar beet fiber, bran fiber, bamboo fiber, wood pulp fiber, cotton fiber, citrus pulp fiber, grass fiber, willow fiber, poplar fiber, cocoa fiber, or combinations thereof. In some embodiments, the filler is derived from wood pulp fiber.

In some embodiments, the filler is a cellulose material. One particularly suitable filler for use in the compositions described herein is microcrystalline cellulose (“MCC”). MCC is typically derived from wood pulp fiber. MCC is composed of glucose units connected by a 1-4 beta glycosidic bond, and may be synthesized by partially depolymerizing alpha-cellulose, by, for example, reactive extrusion, enzyme mediated depolymerization, mechanical grinding, ultrasonication, steam explosion and/or acid hydrolysis. The MCC may be synthetic or semisynthetic, or it may be obtained entirely from natural celluloses. The MCC may be selected from the group consisting of AVICEL® grades PH-100, PH-101, PH-102, PH-103, PH-105, PH-112, PH-113, PH-200, PH-300, PH-301, PH-302, VIVACEL® grades 101, 102, 12, 20 and EMOCEL® grades 50 M and 90 M, and the like, and mixtures thereof. In some embodiments, the oral product comprises MCC as the filler or bulking agent.

In some embodiments, the filler or bulking agent is a non-tobacco plant material or a derivative thereof. Non-limiting examples of derivatives of non-tobacco plant material include starches (e.g., from potato, wheat, rice, corn), natural cellulose, and modified cellulosic materials. Additional examples of potential fillers include maltodextrin, dextrose, calcium carbonate, calcium phosphate, lactose, mannitol, xylitol, and sorbitol. Combinations of these fillers can also be used.

“Starch” as used herein may refer to pure starch from any source, modified starch, or starch derivatives. Starch is present, typically in granular form, in almost all green plants and in various types of plant tissues and organs (e.g., seeds, leaves, rhizomes, roots, tubers, shoots, fruits, grains, and stems). Starch can vary in composition, as well as in granular shape and size. Often, starch from different sources has different chemical and physical characteristics. A specific starch can be selected for inclusion in the composition based on the ability of the starch material to impart a specific organoleptic property to composition. Starches derived from various sources can be used. For example, major sources of starch include cereal grains (e.g., rice, wheat, and maize) and root vegetables (e.g., potatoes and cassava). Other examples of sources of starch include acorns, arrowroot, arracacha, bananas, barley, beans (e.g., favas, lentils, mung beans, peas, chickpeas), breadfruit, buckwheat, canna, chestnuts, colacasia, katakuri, kudzu, malanga, millet, oats, oca, Polynesian arrowroot, sago, sorghum, sweet potato, quinoa, rye, tapioca, taro, tobacco, water chestnuts, and yams. Certain starches are modified starches. A modified starch has undergone one or more structural modifications, often designed to alter its high heat properties. Some starches have been developed by genetic modifications, and are considered to be “modified” starches. Other starches are obtained and subsequently modified. For example, modified starches can be starches that have been subjected to chemical reactions, such as esterification, etherification, oxidation, depolymerization (thinning) by acid catalysis or oxidation in the presence of base, bleaching, transglycosylation and depolymerization (e.g., dextrinization in the presence of a catalyst), cross-linking, enzyme treatment, acetylation, hydroxypropylation, and/or partial hydrolysis. Other starches are modified by heat treatments, such as pregelatinization, dextrinization, and/or cold water swelling processes. Certain modified starches include monostarch phosphate, distarch glycerol, distarch phosphate esterified with sodium trimetaphosphate, phosphate distarch phosphate, acetylated distarch phosphate, starch acetate esterified with acetic anhydride, starch acetate esterified with vinyl acetate, acetylated distarch adipate, acetylated distarch glycerol, hydroxypropyl starch, hydroxypropyl distarch glycerol, and starch sodium octenyl succinate.

The amount of filler or bulking agent can vary, but is typically at least about 5% to about 95% by weight of the oral product, based on the total weight of the oral product. In some embodiments, the filler or bulking agent (such as a cellulose material, such as MCC) may be present in the oral product in an amount of from about 5% to about 95% by weight, such as from about 10% to about 90% by weight, such as from about 15% to about 85% by weight, such as from about 20% to about 80% by weight, such as from about 25% to about 75% by weight, such as from about 30% to about 70% by weight, such as from about 35% to about 65% by weight, such as from about 40% to about 60% by weight of the oral product. In some embodiments, the filler or bulking agent (such as a cellulose material, such as MCC) may be present in an amount of from about 45% to about 55% by weight of the oral product.

In some embodiments, the filler or bulking agent (e.g., cellulose material, such as MCC) may be present in the oral product in an amount of at least about 40% by weight of the oral product, such as at least about 50% by weight of the oral product, such as at least about 55% by weight of the oral product, such as at least about 60% by weight of the oral product. In some embodiments, the cellulose material is present in an amount of at least about 50% by weight of the oral product. In some embodiments, the filler or bulking agent (e.g., cellulose material, such as MCC) may be present in the oral product in an amount of from about 40% to about 99% by weight of the oral product, such as from about 50% to about 95% by weight of the oral product, such as from about 50% to about 90% by weight of the oral product, such as from about 55% to about 85% by weight of the oral product, such as from about 60% to about 80% by weight of the oral product, such as from about 60% to about 75% by weight of the oral product.

In some embodiments, the oral product comprises microcrystalline cellulose in an amount of from about 45% to about 95% by weight of the oral product. In some embodiments, the oral product comprises microcrystalline cellulose in an amount of from about 55% to about 95% by weight of the oral product. In some embodiments, the oral product comprises microcrystalline cellulose in an amount of from about 55% to about 80% by weight of the oral product. In some embodiments, the oral product comprises microcrystalline cellulose in an amount of from about 45% to about 55% by weight of the oral product.

Alternatively or in addition to the above, the at least one filler or bulking agent is or comprises tobacco material. The tobacco material may be included as a bulking agent in the formulation. “Tobacco material” as used herein includes a material which comprises tobacco and/or a tobacco replacement or substitute. In some embodiments, the tobacco material comprises tobacco. In some embodiments, the tobacco material is tobacco. In some embodiments, the tobacco material comprises a tobacco replacement or substitute. In some embodiments, the tobacco material is a tobacco replacement or substitute.

“Tobacco” as used herein includes any part, such as the leaves, flowers, or stems, of any member of the genus Nicotiana and reconstituted materials thereof. In some embodiments, it includes treated tobacco. In some embodiments, it includes derivatives such as specific compounds found in natural tobacco, such as nicotine, whether extract or synthesized, as well as structural derivatives such as the fibrous portion of a tobacco leaf. The term “tobacco” as used herein includes tobacco extract.

The term “tobacco replacement or substitute” as used herein includes tobacco substitutes which comprise individual chemicals and/or complex chemical entities which, when appropriately prepared, physically resemble natural tobacco. Alternatively or in addition, the term “tobacco replacement or substitute” as used herein includes materials which deliver nicotine to the user and provide a similar mouthfeel to tobacco.

When the tobacco comprises plant material, defined amounts of the different parts of the plant may be used. For example, the amount of stem in the tobacco blend may be up to 50%, up to 60%, or up to 70% by weight of the tobacco. In some embodiments, the amount of stem in the tobacco is from 5% to 70% by weight of the tobacco, such as from 10% to 65% by weight of the tobacco, such as from 15% to 65% by weight of the tobacco, such as from 20% to 60% by weight of the tobacco, such as from 25% to 55% by weight of the tobacco, such as from 30% to 50% by weight of the tobacco.

Tobaccos used in the present invention may vary. In some embodiments, the tobacco material is or comprises tobacco such as flue-cured or Virginia (e.g. K326), Burley, sun-cured (e.g., Indian Kurnool and Oriental tobaccos, including Katerini, Prelip, Komotini, Xanthi and Yambol tobaccos), Maryland, dark, dark-fired, dark air-cured (e.g., Passanda, Cubano, Jatin and Bezuki tobaccos), light air-cured (e.g., North Winsconsin and Galpao tobaccos), Indian air-cured, Red Russian and Rustica tobaccos, as well as various other rare or specialty tobaccos and various blends of any of the foregoing tobaccos. Exemplary Nicotiana species include N. tabacum, N. rustica, N. alata, N. arentsii, N. excelsior, N. forgetiana, N. glauca, N. glutinosa, N. gossei, N. Kawakamii, N. knightiana, N. langsdorffi, N. otophora, N. setchelli, N. sylvestris, N. tomentosa, N. tomentosiformis, N. undulata, N. x sanderae, N. Africana, N. amplexicaulis, N. benavidesii, N. bonariensis, N. debneyi, N. longiflora, N. maritiana, N. megalosiphon, N. occidentalis, N. paniculata, N. plumbaginifolia, N. raimondii, N. rosulata, N. simulans, N. stocktonii, N. suaveolens, N. umbratica, N. velutina, N. wigandioides, N. acaulis, N. acuminata, N. attenuata, N. benthamiana, N. cavicola, N. clevelandii, N. cordifolia, N. corymbosa, N. fragrans, N. goodspeedii, N. linearis, N. miersii, N. nudicaulis, N. obtusifolia, N. occidentalis subsp. Hersperis, N. pauciflora, N. petunioides, N. quadrivalvis, N. repanda, N. rotundifolia, N. solanifolia, and N. spegazzinii.

In some embodiments, the tobacco may include types of tobaccos such as dark air-cured tobacco, sun-cured tobacco, flue-cured tobacco, Virginia tobacco, Burley tobacco, Oriental tobacco, Maryland tobacco, dark tobacco, dark-fired tobacco and Rustica tobaccos, as well as other rare or specialty tobaccos.

In some embodiments, the tobacco material is or comprises ground tobacco and/or is or comprises tobacco in particulate form. In some embodiments, the tobacco is not in the form of strands or cut lamina.

Various parts or portions of the plant of the Nicotiana species can be employed. For example, virtually all of the plant (e.g., the whole plant) can be harvested, and employed as such. Alternatively, various parts or pieces of the plant can be harvested or separated for further use after harvest. For example, the flower, leaves, stem, stalk, roots, seeds, and various combinations thereof, can be isolated for further use or treatment. In some embodiments, the tobacco material subjected to the treatments set forth herein is Rustica stems in milled form.

The post-harvest processing of the plant or portion thereof can vary. After harvest, the plant, or portion thereof, can be used in a green form (e.g., the plant or portion thereof can be used without being subjected to any curing process). For example, the plant or portion thereof can be used without being subjected to significant storage, handling or processing conditions. In certain situations, it is advantageous for the plant or portion thereof be used virtually immediately after harvest. Alternatively, for example, a plant or portion thereof in green form can be refrigerated or frozen for later use, freeze dried, subjected to irradiation, yellowed, dried, cured (e.g., using air drying techniques or techniques that employ application of heat), heated or cooked (e.g., roasted, fried or boiled), or otherwise subjected to storage or treatment for later use.

The harvested plant or portion thereof can be physically processed. The plant or portion thereof can be separated into individual parts or pieces (e.g., the leaves can be removed from the stems, and/or the stems and leaves can be removed from the stalk). The harvested plant or individual parts or pieces can be further subdivided into parts or pieces (e.g., the leaves can be shredded, cut, comminuted, pulverized, milled or ground into pieces or parts that can be characterized as filler-type pieces, granules, particulates or fine powders). The plant, or parts thereof, can be subjected to external forces or pressure (e.g., by being pressed or subjected to roll treatment). When carrying out such processing conditions, the plant or portion thereof can have a moisture content that approximates its natural moisture content (e.g., its moisture content immediately upon harvest), a moisture content achieved by adding moisture to the plant or portion thereof, or a moisture content that results from the drying of the plant or portion thereof. For example, powdered, pulverized, ground or milled pieces of plants or portions thereof can have moisture contents of less than about 25 weight percent, such as less than about 20 weight percent, such as less than about 15 weight percent.

Tobacco compositions intended to be used in a smokeless form as described herein may incorporate a single type of tobacco (e.g., in a so-called “straight grade” form). For example, the tobacco within a tobacco composition may be composed solely of flue-cured tobacco (e.g., all of the tobacco may be composed, or derived from, either flue-cured tobacco lamina or a mixture of flue-cured tobacco lamina and flue-cured tobacco stem). In one embodiment, the tobacco comprises or is composed solely of sun-cured milled Rustica stems. The tobacco within a tobacco composition also may have a so-called “blended” form. For example, the tobacco within a tobacco composition of the present invention may include a mixture of parts or pieces of flue-cured, burley (e.g., Malawi burley tobacco) and Oriental tobaccos (e.g., as tobacco composed of, or derived from, tobacco lamina, or a mixture of tobacco lamina and tobacco stem). For example, a representative blend may incorporate about 30 to about 70 parts burley tobacco (e.g., lamina, or lamina and stem), and about 30 to about 70 parts flue cured tobacco (e.g., stem, lamina, or lamina and stem) on a dry weight basis. Other exemplary tobacco blends incorporate about 75 parts flue-cured tobacco, about 15 parts burley tobacco, and about 10 parts Oriental tobacco; or about 65 parts flue-cured tobacco, about 25 parts burley tobacco, and about 10 parts Oriental tobacco; or about 65 parts flue-cured tobacco, about 10 parts burley tobacco, and about 25 parts Oriental tobacco; on a dry weight basis. Other exemplary tobacco blends incorporate about 20 to about 30 parts Oriental tobacco and about 70 to about 80 parts flue-cured tobacco.

The tobacco material can have the form of processed tobacco parts or pieces, cured and aged tobacco in essentially natural lamina and/or stem form, a tobacco extract, extracted tobacco pulp (e.g., using water as a solvent), or a mixture of the foregoing (e.g., a mixture that combines extracted tobacco pulp with granulated cured and aged natural tobacco lamina). The tobacco that is used for the tobacco product may include tobacco lamina, or a tobacco lamina and stem mixture. Portions of the tobaccos within the tobacco product may have processed forms, such as processed tobacco stems (e.g., cut-rolled stems, cut-rolled-expanded sterns or cut-puffed sterns), or volume expanded tobacco (e.g., puffed tobacco, such as dry ice expanded tobacco (DIET)). See, for example, the tobacco expansion processes set forth in U.S. Pat. Nos. 4,340,073 to de la Burde et al.; 5,259,403 to Guy et al.; and 5,908,032 to Poindexter, et al.; and 7,556,047 to Poindexter, et al., all of which are incorporated by reference. In addition, the tobacco product optionally may incorporate tobacco that has been fermented. See, also, the types of tobacco processing techniques set forth in PCT WO 05/063060 to Atchley et al., which is incorporated herein by reference.

The tobacco material may be provided in a shredded, ground, granulated, fine particulate, or powder form. In some embodiments, the tobacco is employed in the form of parts or pieces that have an average particle size less than that of the parts or pieces of shredded tobacco used in so-called “fine cut” tobacco products. Typically, the very finely divided tobacco particles or pieces are sized to pass through a screen of about 18 or 16 Tyler mesh, generally are sized to pass a screen of about 20 Tyler mesh, for example sized to pass through a screen of about 50 Tyler mesh, for example sized to pass through a screen of about 60 Tyler mesh, may even be sized to pass through a screen of 100 Tyler mesh, and further may be sized so as to pass through a screen of 200 Tyler mesh. If desired, air classification equipment may be used to ensure that small sized tobacco particles of the desired sizes, or range of sizes, may be collected. In one embodiment, the tobacco material is in particulate form sized to pass through an 18 or 16 Tyler mesh, but not through a 60 Tyler mesh. If desired, differently sized pieces of granulated tobacco may be mixed together. Typically, the very finely divided tobacco particles or pieces suitable for snus products have a particle size greater than -8 Tyler mesh, such as -8 to +100 Tyler mesh, such as -16 to +60 Tyler mesh. In certain embodiments, the tobacco is provided with an average particle size of about 0.3 to about 2 mm, for example about 0.5 to about 1.5 mm, for example about 0.75 to about 1.25 mm (e.g., about 1 mm).

The manner by which the tobacco is provided in a finely divided or powder type of form may vary. Preferably, tobacco parts or pieces are comminuted, ground or pulverized into a powder type of form using equipment and techniques for grinding, milling, or the like. In some embodiments, the tobacco is relatively dry in form during grinding or milling, using equipment such as hammer mills, cutter heads, air control mills, or the like. For example, tobacco parts or pieces may be ground or milled when the moisture content thereof is less than about 15 weight percent to less than about 5 weight percent.

If included, the tobacco material may be present in an amount of less than about 5% by weight of the oral product, such as no more than about 4% by weight of the oral product, such as no more than about 3% by weight of the oral product, such as no more than about 2% by weight of the oral product, such as no more than about 1% by weight of the oral product, such as no more than about 0.5% by weight of the oral product, such as no more than about 0.1% by weight of the oral product, such as no more than about 0.01% by weight of the oral product.

In some embodiments, the oral product is substantially free from any tobacco material. The term “substantially free from any tobacco material” means that the oral product comprises the tobacco material in an amount of no more than about 1%, such as no more than about 0.5%, such as no more than about 0.1%, such as no more than about 0.01% by weight of the oral product. In some embodiments, the oral product is entirely free from any tobacco material (i.e. the oral product is tobacco-free).

For the avoidance of doubt, combinations of the above end points are explicitly envisaged by the present disclosure. This applies to any of the ranges disclosed herein.

Flavor Glycoside

In addition to the at least one filler or bulking agent, the oral product comprises at least one flavor glycoside. As used herein, the term “flavor glycoside” refers to a compound in which a flavoring agent is bound to a sugar molecule via a glycosidic bond.

The glycosidic bond may be an O—, N—, S— or C-glycosidic bond. An O-glycosidic bond is formed between the anomeric carbon on the sugar and a hydroxyl group on the flavoring agent. An N-glycosidic bond is formed between the anomeric carbon on the sugar and an amino group on the flavoring agent.

Flavor glycosides as used herein may be prepared using any methods known to the person skilled in the art. For example, the flavor glycoside may be produced by synthetic (i.e. chemical) methods or may be produced by a biotechnological process. As the skilled person is aware, industrial production of glycosides may carried out by the Koenigs-Knorr process (i.e. organic-chemical substitution of a glycosyl halide with an alcohol to yield a glycoside) or reversed enzymatic hydrolysis or transglycosylation employing glycosidases. The flavor glycoside may be free from compounds originating from an extract of a natural plant.

In some embodiments, the flavor glycoside is not produced by synthetic or chemical methods.

Biotechnological Process for Preparing Flavor Glycoside

In some embodiments, the at least one flavor glycoside may be obtained from a biotechnological process. The biotechnological process may be an enzymatic process of producing the flavor glycoside, such as a process involving a glycosyltransferase.

The flavor glycoside obtained from a biotechnological process may be a flavor glycoside that is commercially available from 4Gene GmbH. For example, the flavor glycoside obtained from a biotechnological process may be a flavor glucoside that is commercially available from 4Gene GmbH.

The biotechnological process that may be used to prepare the flavor glycoside may be as described in WO 2015/197844, the entirety of which is hereby expressly incorporated by reference. The flavor glycoside may be prepared by contacting a flavoring agent with a sugar donor and a glycosyl transferase under conditions appropriate for the transfer of the sugar group of the sugar donor to a hydroxyl group, or other suitable functional group, on the flavoring agent. The glycosyl transferase may be a recombinantly expressed glycosyl transferase.

As the skilled person will appreciate, in embodiments wherein the flavor glycoside comprises a sugar that is a disaccharide, oligosaccharide or polysaccharide, the enzymatic process may include the same steps as for a monosaccharide, but in which the steps are repeated the corresponding number of times as for the number of saccharide units. For example, whilst the preparation of a flavor glycoside in which the sugar is a monosaccharide may include the above-mentioned steps, the preparation of a flavor glycoside in which the sugar is a disaccharide may comprise the same steps as for the monosaccharide but in which these are repeated twice.

For example, the methods described in Example 1 of WO 2015/197844, the entirety of which is hereby incorporated, can be used to produce the flavor glycoside as described herein.

The glycosyl transferase may have an amino acid sequence that (a) comprises the sequence of SEQ ID NO: 1; or (b) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to SEQ ID NO: 1; or (c) comprises a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (d) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (e) comprises the sequence of SEQ ID NO: 2; or (f) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to SEQ ID NO: 2; or (g) comprises a part of the sequence of SEQ ID NO: 2, wherein, preferably, said part of the sequence of SEQ ID NO: 2 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (h) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to a part of the sequence of SEQ ID NO: 2, wherein, preferably, said part of the sequence of SEQ ID NO: 2 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length.

The glycosyl transferase may be a small molecule glycosyl transferase. In some embodiments, the glycosyl transferase is a terpene glycosyl transferase, preferably a monoterpene glycosyl transferase, more preferably a UDP-glucose mono terpene β-D-glucosyltransferase. In some embodiments, said glycosyl transferase is capable of using UDP-glucose as sugar donor. Preferably, said glycosyl transferase uses UDP-glucose more efficiently as sugar donor than UDP-xylose, UDP -glucuronic acid, UDP-arabinose, UDP-rhamnose, UDP-galactose, GDP-fucose, GDP-mannose and/or CMP-sialic acid, as seen by radiochemical analysis. In such radiochemical analysis, individual reactions are carried out in which different radiolabeled sugar donors (such as radiolabeled UDP-glucose, UDP-xylose and UDP- glucuronic acid) that carry a radionuclide in their sugar group are reacted under appropriate conditions and in the presence of the glycosyl transferase with a certain acceptor molecule. By comparing the amount of radiolabel that was transferred from the different sugar donors to the acceptor molecule, it can be determined which sugar donor the glycosyl transferase uses more efficiently than the others.

In some embodiments, the glycosyl transferase is capable of catalyzing transfer of a sugar group from a sugar donor to a hydroxyl group of a hydroxy-containing terpene and/or a carboxyl group of a carboxy-containing terpene. In some embodiments, the glycosyl transferase is capable of catalyzing formation of a glycoside in which a sugar is linked to a hydroxy-containing terpene through a β-D-glycosyl linkage and/or formation of a glycose ester in which a sugar is linked to a carboxy-containing terpene through a β-D-glycose ester linkage.

In some embodiments, the glycosyl transferase is capable of catalyzing glycosylation, preferably glucosylation, of geraniol, (R-)linalool, (R- and/or S-)citronellol, nerol, hexanol and/or octanol, preferably geraniol and/or (R- and/or S-)citronellol, wherein, preferably, said glycosyl transferase has an amino acid sequence as defined above in (a) to (d). Namely, said glycosyl transferase has an amino acid sequence that (a) comprises the sequence of SEQ ID NO: 1 ; or (b) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to SEQ ID NO: 1; or (c) comprises a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (d) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length.

In some embodiments, the glycosyl transferase is capable of catalyzing glycosylation, preferably glucosylation, of furaneol, wherein, preferably, said glycosyl transferase has an amino acid sequence as defined above in (a) to (d), i.e. that (a) comprises the sequence of SEQ ID NO: 1; or (b) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to SEQ ID NO: 1; or (c) comprises a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (d) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%), identical to a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length.

The glycosyl transferase may be capable of catalyzing glycosylation, preferably glucosylation, of eugenol, wherein, preferably, said glycosyl transferase has an amino acid sequence as defined above in (a) to (d), i.e. that (a) comprises the sequence of SEQ ID NO: 1 ; or (b) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to SEQ ID NO: 1 ; or (c) comprises a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (d) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to a part of the sequence of SEQ ID NO: 1, wherein, preferably, said part of the sequence of SEQ ID NO: 1 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length.

In some embodiments, the glycosyl transferase is capable of catalyzing glycosylation, preferably glucosylation, of geraniol, (R- and/or S-)citronellol, nerol, hexanol, octanol, 8-hydroxylinalool, trans 2-hexenol, and/or farnesol, preferably geraniol, wherein, preferably, said glycosyl transferase has an amino acid sequence as defined above in (d) to (h), i.e. that (d) comprises a sequence that is at least 90%), preferably at least 95%, more preferably at least 98%, identical to a part of the sequence of SEQ ID NO: 2, wherein, preferably, said part of the sequence of SEQ ID NO: 2 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; (e) comprises the sequence of SEQ ID NO: 2; or (f) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to SEQ ID NO: 2; or (g) comprises a part of the sequence of SEQ ID NO: 2, wherein, preferably, said part of the sequence of SEQ ID NO: 2 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length; or (h) comprises a sequence that is at least 90%, preferably at least 95%, more preferably at least 98%, identical to a part of the sequence of SEQ ID NO: 2, wherein, preferably, said part of the sequence of SEQ ID NO: 2 is at least 50, preferably at least 80, more preferably at least 100, more preferably at least 200, amino acids in length.

Also described herein is an (isolated) nucleic acid molecule encoding a glycosyl transferase as defined in any of the embodiments described above, wherein, preferably, said nucleic acid molecule is a DNA molecule.

Also described herein is a vector comprising a DNA sequence encoding a glycosyl transferase as defined in any of the embodiments described above. The vector may be an expression vector, preferably an expression vector for expression of a glycosyl transferase as defined in any of the embodiments described above.

Also described herein is a host cell containing or transfected with the nucleic acid molecule as described above or the vector as described above, wherein, preferably, said host cell is not a cell of Vitis vinifera, more preferably not a cell of a grape vine, and/or wherein, preferably, said host cell is a non-human cell, preferably a bacterial cell, more preferably an E.coli cell. In some embodiments, said host cell produces/expresses a glycosyl transferase as defined in any of the embodiments above.

Also described herein is a transgenic plant comprising a nucleic acid molecule as defined above or a vector as defined above, wherein, preferably, said plant is not a Vitis vinifera plant, more preferably not a grape vine. In some embodiments, said transgenic plant produces/expresses a glycosyl transferase as defined in any of the embodiments above.

As described herein, the glycosyl transferase as defined in any of the embodiments above or a nucleic acid molecule as defined above or a vector as defined above or a host cell as defined above or a transgenic plant as defined above may be used for producing a flavor glycoside, such as for example a terpene glycoside, an octanyl glycoside, furaneyl glycoside or hexanyl glycoside.

The production of said flavor glycoside may not involve steps carried out in vivo. In some embodiments, said production of the flavor glycoside is carried out in a host cell or transgenic plant as defined above, preferably in an E.coli cell. In some embodiments, the sugar group of the flavor glycoside is glucose and the sugar donor used in the method of production is UDP-glucose,

In some embodiments, the method for producing the flavor glycoside is an in vitro method which does not involve any steps carried out in vivo. In some embodiments, said method is an in vivo method carried out in a host cell or transgenic plant. In some embodiments, said method may be an in vivo method carried out in E. coli.

In some embodiments, the biotechnological method of obtaining the flavor glycoside may comprise the steps of:

culturing or growing a host cell as defined above or a transgenic plant as defined above; and
collecting from said host cell or transgenic plant the flavor glycoside.

The flavor glycoside may be a flavor glycoside in which a hydroxy-containing flavoring agent is covalently linked to a sugar group. In some embodiments, during said culturing or growing said host cell or transgenic plant said hydroxy- containing flavoring agent is present in said host cell or transgenic plant. In some embodiments, during said culturing or growing said host cell or transgenic plant UDP-glucose is present in the culture medium used for culturing said host cell or in the water used for watering that transgenic plant.

In some embodiments, said culturing or growing said host cell is carried out in a bioreactor. A “bioreactor” is a vessel in which a (bio)chemical process is carried out which involves organisms (such as host cells) or biochemically active substances derived from such organisms.

Also described herein is a method of producing a protein having glycosyl transferase activity and/or enzymatic activity for the catalysis of glycose esterification, said method comprising the steps of:

culturing or growing a host cell as defined above or a transgenic plant as above; and, preferably,
collecting from the host cell or transgenic plant a protein having glycosyl transferase activity and/or enzymatic activity for the catalysis of glycose esterification.

In some embodiments, said glycosyl transferase activity is an activity of transferring the sugar group of a sugar donor to a hydroxyl group of a hydroxy-containing flavoring agent under formation of a glycosidic bond between said hydroxy-containing flavoring agent and said sugar group.

The protein having glycosyl transferase activity and/or enzymatic activity for the catalysis of glycose esterification may be a glycosyl transferase as defined in any of the embodiments above. A “glycosyl transferase” in an enzyme of EC class 2.4 that catalyzes the transfer of a monosaccharide moiety from a sugar donor to an acceptor molecule under formation of a glycosidic linkage between the sugar (the glycone) and the acceptor molecule (the aglycone) (see, e.g., Bowles et al., 2006). The sugar donor may be an activated sugar precursor and can be, for example, UDP(uridine diphosphate)-glucose wherein the sugar is glucose, UDP -xylose wherein the sugar is xylose, UDP-glucuronic acid wherein the sugar is glucuronic acid, UDP-arabinose wherein the sugar is arabinose, UDP-rhamnose wherein the sugar is rhamnose, UDP-galactose wherein the sugar is galactose, GDP(guanosin diphosphate)-fucose wherein the sugar is fucose, GDP-mannose wherein the sugar is mannose or CMP(cytidine monophosphate)-sialic acid wherein the sugar is sialic acid. If the glycosyl transferase is a glucosyl transferase, then the sugar donor is UDP-glucose.

The acceptor molecule may be an alcohol (OH group), such as the alcohol of a terpenoid, alkaloid, cyanohydrin, glucosinolate, flavonoid, isoflavonoid, anthocyanidin, phenylpropanoid, polyphenol, hydroquinone, amine, carbohydrate (monomeric or oligomeric), fatty acid or lipids. Examples of glycosyl transferases are UDP-glucosyltransferases, UDP-arabinosyltransferases, UDP- glucuronosyltransferases, UDP-xylosyltransferases, UDP-galactosyltransferases, UDP- rhamnosyltransferases, GDP-fucosyltransferase, GDP-mannosyltransferase, or CMP- sialyltransferase. A glycosyl transferase may or may not have an additional enzymatic activity for the catalysis of glycose esterification, i.e. for transferring the sugar group of a sugar donor to a carboxyl group of a carboxy-containing acceptor molecule under formation of a glycose ester bond between said carboxy-containing acceptor molecule and said sugar group.

A “small molecule glycosyl transferase” is a glycosyl transferase that catalyzes the transfer of a monosaccharide moiety from a sugar donor to a small molecule as acceptor molecule. A small molecule is a molecule that has a molecular weight below 1 500 Dalton, preferably below 1 000 Dalton. A “flavoring agent glycosyl transferase” is a glycosyl transferase that catalyzes the transfer of a monosaccharide moiety from a sugar donor to a flavoring agent as acceptor molecule. A “terpene glycosyl transferase” is a glycosyl transferase that catalyzes the transfer of a monosaccharide moiety from a sugar donor to a terpene as acceptor molecule. A “monoterpene glycosyl transferase” is a glycosyl transferase that catalyzes the transfer of a monosaccharide moiety from a sugar donor to a monoterpene as acceptor molecule. An “UDP-glucose:monoterpene β-D-glucosyltransferase” is a glycosyl transferase that catalyzes the transfer of a glucose moiety from a UDP-glucose as sugar donor to a monoterpene as acceptor molecule under formation of covalent a β-D-glycosidic bond.

Reference is made herein to a glycosyl transferase “having” a certain amino acid sequence. This is meant to designate that the amino acid sequence of said glycosyl transferase consists of said certain amino acid sequence, i.e. the glycosyl transferase has only said certain amino acid sequence and no further amino acid sequence(s) beyond said certain amino acid sequence.

Glycosyl transferases having an amino acid sequence comprising SEQ ID NO: 1 (i.e. the sequence of VvGT14) or SEQ ID NO: 2 (i.e. the sequence of VvGT15) or a related amino acid sequence can be obtained by standard methods of recombinant DNA technology, for example as described in Example 1 of WO 2015/197844, which is hereby incorporated by reference in its entirety.

A “glycoside”, as used herein, is a molecule in which a sugar (the “glycone” part or “glycone component” of the glycoside) is bonded to a non-sugar (the “aglycone” part or “aglycone component”) via a glycosidic bond. Accordingly, a glycoside may consist of a sugar as glycone component (designated “Z” in the general chemical structure below) linked through its anomeric carbon atom to the hydroxy group of an alcohol (chemical structure R-OH) as aglycone component, thus resulting in a glycoside of the general chemical structure R-O-Z. For example, in the glycoside linaloyl β-D-glucoside, the glycone component glucose is linked to the aglycone component linalool.

A glycoside can be produced by carrying out a reaction in which an aglycone component (such as a terpene, for example geraniol or citronellol) is mixed under appropriate conditions with a sugar donor (an activated sugar precursor such as UDP-glucose or UDP-glucuronic acid, preferably UDP-glucose) in the presence of a glycosyl transferase as enzymatic catalyzer. For example, 100 µL purified enzyme (50 µg), 100-150 µL Tris-HCl buffer (100 mM, pH 7.5, 10 mM 2-mercaptoethanol), 37 pmol UDP-glucose and 50 µg substrate (dissolved in methyl-tert-butylether) can be incubated at 30° C. for 24 hr. This results in the formation of glycosides composed of an aglycone component linked to a glycone component. The glycoside can subsequently be isolated from the reaction mixture by standard methods of extraction and chromatography (see also Example 1 of WO 2015/197844, which is hereby incorporated by reference in its entirety).

Alternatively, a glycoside can be produced by culturing or growing a host cell or transgenic plant expressing a glycosyl transferase as described herein. During culture/growth, such a host cell or transgenic plant will generate glycosides. The glycoside(s) generated in such a host cell or transgenic plant can subsequently be collected from said host cell or transgenic plant by standard methods of extraction and/or chromatography (such as solvent extraction, solid phase extraction and reversed phase chromatography). In such a method for producing a glycoside, the present disclosure may indicate that during said culturing or growing a host cell or transgenic plant a certain compound or substrate (such as the aglycone component) used for formation of the glycoside “is present in said host cell or transgenic plant”. This means that the compound or substrate is either produced by said host cell or transgenic plant, such that it is present in said host cell/in the cells of said transgenic plant, or that it is added to the host cell or transgenic plant in such a manner that it is taken up by the host cell or transgenic plant and enters into the host cell/cells of the transgenic plant. This may, for example, be achieved by including the compound or substrate to the culture medium used for culturing the host cells (for example the growth medium used for culturing E.coli cells) or, in the case of a transgenic plant, by adding the compound or substrate to the water used for watering the plant (for example, an aqueous solution containing the compound/substrate or an aqueous solution with a low content of ethanol containing the compound/substrate may be added to the culture medium used for culturing the host cells or to the water used for watering the plant).

Further information on a biotechnological process to produce a flavor glycoside is set out in WO 2015/197844.

If the present application refers to “collecting” a certain glycoside, glycose ester or protein from a host cell or transgenic plant, this is meant to designate that said glycoside, glycose ester or protein is separated and/or isolated from other components of said host cell or transgenic plant. This can be achieved by standard methods of extraction and chromatography known to a person of skill in the art (see e.g. Example 1 of WO 2015/197844, which is hereby incorporated by reference in its entirety).

At some instances, the present application refers to a glycosyl transferase being “capable of catalyzing” a certain reaction. For example, the present application may state that a glycosyl transferase is capable of catalyzing transfer of a sugar group from a sugar donor to a certain acceptor. This is meant to designate that under appropriate reaction conditions the rate at which the reaction product (in the example the adduct of the sugar group and the acceptor) is formed is at least 10-fold higher in the presence of said glycosyl transferase than the rate at which the reaction product is formed in a control experiment in the absence of said glycosyl transferase.

At some instances, the present application indicates that a certain glycosyl transferase “has a glucosyl transferase activity” for a substrate A that is “by at least a factor X higher” than the glucosyl transferase activity for a substrate B. This means that, if the k_cat/K_M values (i.e. the specificity constants) of said glycosyl transferase for substrate A and B are measured under appropriate conditions and the k_cat/K_M value obtained for the glycosyl transferase with substrate A is divided by the k_cat/K_M value obtained for the glycosyl transferase with substrate B, the resulting value is X or greater than X.

Similarly, the present application may indicate that a certain glycosyl transferase G “has a glucosyl transferase activity” for a certain substrate A that is “by at least a factor X higher” than the glucosyl transferase activity of another glycosyl transferase H for substrate A. This means that, if the k_cat/K_M values of glycosyl transferase G and of glycosyl transferase H for substrate A are measured under appropriate conditions and the k_cat/K_M value obtained for glycosyl transferase G with substrate A is divided by the k_cat/K_M value obtained for glycosyl transferase H with substrate A, the resulting value is X or greater than X.

The k_cat/K_M value can be determined by standard procedures known to the person of skilled in the art. Preferably, recombinant glycosyl transferases are used for determining the k_cat/K_m values.

Preferably, the following procedure is used: The kinetic data are determined with increasing concentrations of the substrates from 1 µM to 500 µM and a fixed concentration of sugar precursor (for example an UDP-glucose concentration of 108 µM (100 µM unlabeled UDP-glucose and 8 µM UDP-[¹⁴C] glucose), 833 µM (825 µM unlabeled UDP-glucose and 8 µM UDP-[¹⁴C] glucose) or 512.5 µM (500 µM unlabeled UDP-glucose and 12,5 µM UDP-[¹⁴C] glucose)). The total volume is 40 µL and 0.2 µg, 0.5 µg or 5 µg of purified protein is used. The measurements are performed under the following conditions: The assays are carried out at 30° C. for 1.5 h, 30 min or 10 min using a Tris-HCl buffer (100 mM, 10 mM 2-mercaptoethanol, pH 8.5 or pH 7.5). The amount of the purified enzyme and the incubation time can be adapted depending on the counting sensibility. The reaction is stopped by adding 1 µL 24% trichloroacetic acid and glucosides are extracted with 100 µL ethyl acetate. Radioactivity is determined by LSC.

To determine the kinetic data of a sugar precursor (e.g. UDP-glucose), the value of the substrate used (e.g. geraniol) is fixed (1.25 mM or 0.1 mM) and radiolabeled sugar precursor (e.g. UDP-[¹⁴C] glucose) is mixed with non-radiolabeled sugar precursor (in the example UDP-glucose) to obtain concentrations ranging from 5 µM to 100 µM or 25 µM to 500 µM. The K_M-and V_max- values are calculated from Lineweaver-Burk plots, Hanes-Woolf plots and non-linear fitting of the experimental data.

At some instances, the present application indicates that a certain glycosyl transferase can be “expressed more efficiently as a recombinant protein in E.coli cells” than another glycosyl transferase. The efficiency of recombinant protein expression in E.coli can be compared as follows: Recombinant expression of the different glycosyl transferases is carried out in E.coli cells by standard methods known to the skilled person, preferably according to the methods described in Example 1 of WO 2015/197844, which is hereby incorporated by reference in its entirety. Whole-cell extracts from the E.coli cells are prepared and proteins in the whole-cell extract are compared after gel-electrophoresis and visualization by coomassie-staining.

As “host cell” transfected with the nucleic acid molecule as described above, the cell of a prokaryotic or eukaryotic organism may be used. As the prokaryotic organism, bacteria, for example, commonly used hosts such as bacteria belonging to genus Escherichia such as Escherichia coli can be used. Alternatively, a cell of a lower eukaryotic organism such as eukaryotic microorganisms including, for example, yeast (e.g. Saccharomyces cerevisiae) or fungi like Aspergillus oryzae and Aspergillus niger can be used. Animal cells or plant cells also can be used as a host. Examples of animal cells that can be used include cell lines of mouse, hamster, monkey, human, etc., as well as insect cells such as silkworm cells and adult silkworm per se.

Construction of a vector may be performed using a restriction enzyme, ligase etc. according to a standard method known in the art. An “expression vector” is a vector that allows expression of a protein encoded by the DNA sequence of the vector in a target cell. The transformation of a host with an (expression) vector can be performed according to standard methods.

At some instances, the present application refers to a host cell being “transfected”. This refers to a situation where foreign DNA is introduced into a cell. A transfected host cell may be “stably transfected”. This refers to the introduction and integration of foreign DNA into the genome of the transfected cell. Alternatively, a transfected host cell may be “transiently transfected”. This refers to the introduction of foreign DNA into a cell where the foreign DNA fails to integrate into the genome of the transfected cell.

As used herein, the term “transgenic plant” refers to a plant that has a heterologous gene integrated into its genome and that transmits said heterologous gene to its progeny. A “heterologous gene” is a gene that is not in its natural environment. For example, a heterologous gene includes a gene from one species introduced into another species. In some embodiments, a heterologous gene also includes a gene native to an organism that has been altered in some way (e.g., mutated, added in multiple copies, or linked to non-native regulatory sequences). Heterologous genes are distinguished from endogenous genes in that the heterologous gene sequences are typically joined to DNA sequences that are not found naturally associated with the gene sequences in the chromosome or are associated with portions of the chromosome not found in nature (e.g., genes expressed in loci where the gene is not normally expressed).

A “protein having glycosyl transferase activity” is a protein that is capable of catalyzing a glycosylation reaction in which the sugar group of a sugar donor is transferred to an acceptor molecule. A protein having glycosyl transferase activity can be obtained by culturing, cultivating or growing a host cell or organism that expresses such a protein (for example a host cell transformed with a vector as described in the above embodiments), and then by recovering and/or purifying the protein from the host cell, host organism or culture medium according to standard methods, such as filtration, centrifugation, cell disruption, gel filtration chromatography, ion exchange chromatography and the like. A “recombinantly expressed” glycosyl transferase is a glycosyl transferase protein that has been expressed from a recombinant DNA molecule, i.e. from a DNA molecule formed by laboratory methods of genetic engineering (such as molecular cloning) to bring together genetic material from multiple sources, creating a DNA sequence that would not be found naturally in a biological organism. Typically, a recombinantly expressed glycosyl transferase is expressed by heterologous expression (i.e. in a host organism which is different from the organism from which said glycosyl transferase is originally derived), such as by expression in e.g. E. coli,Saccharomyces cerevisiae, Pichia pastoris or insect cells, preferably in E. coli. Preferably, said recombinantly expressed glycosyl transferase is expressed by heterologous expression. Preferably, said recombinantly expressed glycosyl transferase is isolated after expression from other proteins of the host organism by methods of protein purification.

The term “reaction product composition”, as used herein, refers to a composition obtained from a method for forming/producing said reaction product upon completion of the reaction step in which said reaction product is actually formed, wherein said composition is not subjected to any further steps of purifying or separating the components of the reaction mixture obtained after said reaction step in which said reaction product is actually formed. If used in the context of a method to produce a product in a host cell or transgenic plant, the term “reaction product composition” refers to the culture supernatant, host cell extract or transgenic plant extract in which said product is harvested from said host cell or transgenic plant. It has been found that WGT14 and VvGT15 have glucosyl transferase activities (k_cat/K_M) for the substrates geraniol, nerol and citronellol that are higher by a factor of 2.6 to 44 compared to known terpene glycosyl transferases, such as UGT85B1 of Sorghum bicolor. Moreover, it has been found that the glycosyl transferases VvGT14 and VvGT15 are expressed more efficiently than other known terpene glycosyl transferases as recombinant proteins in E.coli cells or other host cells. Moreover, it has been found that the glycosyl transferase VvGT14 is capable of catalyzing glucosylation of furaneol, whereas plant glycosyl transferases that are capable of catalyzing glucosylation of furaneol are otherwise not known.

For example, the methods described in Example 1 of WO 2015/197844, the entirety of which is hereby incorporated, can be used to produce the flavor glycoside as described herein.

It has been found that a flavor glycoside obtained from a biotechnological process as described hereinabove may comprise fewer impurities than a flavor glycoside obtained using chemical or synthetic methods. For example, a flavor glycoside prepared using chemical methods may typically contain impurities from e.g. the catalyst used during the synthesis, such as metal impurities. The flavor glycoside obtained from a biotechnological process may comprise less than 0.01 wt% of chemical impurities, such as less than 0.001 wt% of chemical impurities, such as less than 0.0001 wt% of chemical impurities. The flavor glycoside obtained from a biotechnological process may be entirely free from chemical impurities, such as entirely free from metallic impurities.

Characteristics of Flavor Glycoside

The identity of the sugar molecule that forms the backbone of the flavor glycoside may vary and may be any suitable saccharide. As the skilled person will appreciate, any saccharide in which the anomeric carbon is free (i.e. the anomeric carbon is not already bound with another molecule) will be suitable for forming a glycosidic bond. In some embodiments, the sugar is a monosaccharide or disaccharide. In some embodiments, the sugar is a monosaccharide; the monosaccharide may be a pentose or a hexose. In some embodiments, the sugar is a pentose. In some embodiments the sugar is a pentose selected from the group consisting of ribose, deoxy-ribose, xylose and arabinose. In some embodiments, the sugar is a hexose. In some embodiments, the sugar is a hexose selected from the group consisting of glucose, fructose and galactose. In some embodiments, the sugar is a monosaccharide selected from the group consisting of glucose and fructose.

In some embodiments, the sugar is a diglycoside. As used herein, the term diglycoside refers to a compound having two glycoside groups; i.e. a sugar which can form two glycosidic bonds with one or two flavoring agents. In some embodiments, the sugar is a diglycoside selected from the group consisting of alpha-L-arabinofuranose, alpha-L-rhamnopyranose, beta-D-glucopyranose, beta-D-apiofuranose and beta-D-xylopyranose.

In some embodiments, the sugar is glucose and the flavor glycoside is a flavor glucoside. In some embodiments, the sugar is fructose.

As used herein, the term “flavoring agent” (or “flavor” or “flavorant”) refers to materials which, where local regulations permit, may be used to create a desired taste, aroma or other somatosensorial sensation in a product for adult consumers. Examples of sensory characteristics that can be modified by the flavoring agent include taste, mouthfeel, moistness, coolness/heat, and/or fragrance/aroma. Flavoring agents may be natural or synthetic, and the character of the flavors imparted thereby may be described, without limitation, as fresh, sweet, herbal, confectionary, floral, fruity, or spicy.

The flavoring agent bound via a glycosidic bond to the sugar molecule in the flavor glycoside may vary. As the skilled person will appreciate, any flavoring agent having a hydroxyl group or amino group may be a suitable flavoring agent for forming a flavor glycoside.

The flavoring agent may be selected from the group consisting of naturally occurring flavor materials, botanicals, extracts of botanicals, synthetically obtained materials, or combinations thereof (e.g., tobacco, cannabis, licorice (liquorice), hydrangea, eugenol, Japanese white bark magnolia leaf, chamomile, fenugreek, clove, maple, matcha, menthol, Japanese mint, aniseed (anise), cinnamon, turmeric, Indian spices, Asian spices, herb, wintergreen, cherry, berry, red berry, cranberry, peach, apple, orange, mango, clementine, lemon, lime, tropical fruit, papaya, rhubarb, grape, durian, dragon fruit, cucumber, blueberry, mulberry, citrus fruits, Drambuie, bourbon, scotch, whiskey, gin, tequila, rum, spearmint, peppermint, lavender, aloe vera, cardamom, celery, cascarilla, nutmeg, sandalwood, bergamot, geranium, khat, naswar, betel, shisha, pine, honey essence, rose oil, vanilla, lemon oil, orange oil, orange blossom, cherry blossom, cassia, caraway, cognac, jasmine, ylang-ylang, sage, fennel, wasabi, piment, ginger, coriander, coffee, hemp, a mint oil from any species of the genus Mentha, eucalyptus, star anise, cocoa, lemongrass, rooibos, flax, ginkgo biloba, hazel, hibiscus, laurel, mate, orange skin, rose, tea such as green tea or black tea, thyme, juniper, elderflower, basil, bay leaves, cumin, oregano, paprika, rosemary, saffron, lemon peel, mint, beefsteak plant, curcuma, cilantro, myrtle, cassis, valerian, pimento, mace, damien, marjoram, olive, lemon balm, lemon basil, chive, carvi, verbena, tarragon, limonene, thymol, camphene), flavor enhancers, bitterness receptor site blockers, sensorial receptor site activators or stimulators, sugars and/or sugar substitutes (e.g., sucralose, acesulfame potassium, aspartame, saccharine, cyclamates, lactose, sucrose, glucose, fructose, sorbitol, or mannitol), and other additives such as charcoal, chlorophyll, minerals, botanicals, or breath freshening agents. They may be imitation, synthetic or natural ingredients or blends thereof. They may be in any suitable form, for example, liquid such as an oil, solid such as a powder, or gas.

In some embodiments, the flavoring agent comprises menthol, spearmint and/or peppermint. In some embodiments, the flavoring agent comprises flavor components of cucumber, blueberry, citrus fruits and/or redberry. In some embodiments, the flavoring agent comprises eugenol. In some embodiments, the flavoring agent comprises flavor components extracted from tobacco. In some embodiments, the flavoring agent comprises flavor components extracted from cannabis.

In some embodiments, the flavoring agent may comprise a sensate, which is intended to achieve a somatosensorial sensation which are usually chemically induced and perceived by the stimulation of the fifth cranial nerve (trigeminal nerve), in addition to or in place of aroma or taste nerves, and these may include agents providing heating, cooling, tingling, numbing effect. A suitable heat effect agent may be, but is not limited to, vanillyl ethyl ether and a suitable cooling agent may be, but not limited to eucalyptol, WS-3.

In some embodiments, the flavoring agent is lipophilic. Without wishing to be bound by theory, formulation of a lipophilic flavoring agent as an emulsion may enhance the stability of the flavoring agent (e.g., toward oxidation or evaporation). In some embodiments, the flavoring agent is susceptible to oxidation, meaning exposure to air results in the degradation of components in the flavoring agent due to chemical changes. Examples of functional groups which may be present in flavoring agent components exhibiting susceptibility to oxidation include, but are not limited to, alkenes, aldehydes, and/or ketones. In some embodiments, the flavoring agent comprises a citrus oil. Citrus oils contain, for example, terpene components which may be susceptible to oxidation, evaporation, or both and, thus, may particularly benefit from inclusion within a product in the form of an emulsion as provided herein.

In some embodiments, the flavoring agent may comprise a terpene. In some embodiments, the flavoring agent may comprise a monoterpene and/or a diterpene and/or a sesquiterpene. In some embodiments, the flavoring agent may comprise a monoterpene. A “terpene”, as used herein, is a hydrocarbon having a carbon skeleton formally derived by combination of several isoprene units. The term includes hydrocarbons having a carbon skeleton formally derived by combination of several isoprene units covalently linked to at least one hydroxy group, preferably covalently linked to one hydroxy group and/or covalently linked to at least one carboxyl group, preferably covalently linked to one carboxyl group. In some embodiments, the term “terpene” also includes hydrocarbons having a carbon skeleton formally derived by combination of several isoprene units in which up to three, preferably up to two, more preferably one, methyl groups have been moved or removed. As used herein, a “hydroxy-containing terpene” is a terpene that comprises one or more, preferably one, hydroxy group. The term “terpene glycoside” refers to a glycoside the aglycone component of which is a terpene. The term “monoterpene glycoside” refers to a glycoside the aglycone component of which is a monoterpene (formally comprising two isoprene units, such as geraniol, citronellol or linalool). The term “sesquiteipene glycoside” refers to a glycoside the aglycone component of which is a terpene formally comprising three isoprene units (such as farnesol). The term “diterpene glycoside” refers to a glycoside the aglycone component of which is a diterpene (formally comprising four isoprene units, such as steviol).

In some embodiments, the terpene is a terpene derivable from a phytocannabinoid producing plant, such as a plant from the stain of the cannabis sativa species, such as hemp. Suitable terpenes in this regard include so-called “C10” terpenes, which are those terpenes comprising 10 carbon atoms, and so-called “C15” terpenes, which are those terpenes comprising 15 carbon atoms. In some embodiments, the oral product comprises more than one terpene. For example, the oral product may comprise one, two, three, four, five, six, seven, eight, nine, ten or more terpenes as defined herein. In some embodiments, the terpene is selected from pinene (alpha and beta), geraniol, linalool, limonene, carvone, eucalyptol, menthone, iso-menthone, piperitone, myrcene, beta-bourbonene, germacrene, thymol, citral, eugenol, and mixtures thereof.

In some embodiments, the flavoring agent is selected from the group consisting of terpenes, aliphatic alcohols, aromatic alcohols, pyrones, lactones, or phenylpropanoids, and combinations thereof. In some embodiments, the flavoring agent is or comprises a terpene.

In some embodiments, the flavoring agent is selected from the group consisting of geraniol, citronellol, nerol, maltol, ethylmaltol, fenchol, homofuraneol, furaneol, norfuraneol, 1-octen-3-ol, borneol, linalool, farnesol, hydroxycitronellol, 3,7-dimethyloctanol, myrcenol, lavandulol, nerolidol, terpineol, alpha-terpineol, menthol, thymol, carvacrol, myrtenol, carveol, santalol, piperitol, perillyl alcohol, patchouli alcohol, hexanol, 1-hexanol, 3-cis-hexanol, cis-3-hexen-1-ol, phenylethanol, eugenol, sesamol, sotolone, maple furanone, methyl anthranilate, guaiacol, raspberry ketone, 2-methoxy-4-vinylphenol, 4-ethylguaiacol, benzylalcohol, homofuraneol, vanillin, ethylvanillin, and combinations thereof. In some embodiments, the flavoring agent is selected from the group consisting of geraniol, citronellol, nerol, maltol, ethylmaltol, fenchol, homofuraneol, furaneol, norfuraneol, 1-octen-3-ol, borneol, linalool, farnesol, hydroxycitronellol, 3,7-dimethyloctanol, myrcenol, lavandulol, nerolidol, terpineol, alpha-terpineol, menthol, thymol, carvacrol, myrtenol, carveol, santalol, piperitol, perillyl alcohol, patchouli alcohol, hexanol, 1-hexanol, 3-cis-hexanol, cis-3-hexen-1-ol, phenylethanol, eugenol, sesamol, sotolone, maple furanone, methyl anthranilate, guaiacol, raspberry ketone, 2-methoxy-4-vinylphenol, 4-ethylguaiacol, benzylalcohol, homofuraneol, and combinations thereof.

In some embodiments, the flavoring agent is or comprises raspberry ketone. In some embodiments, the flavoring agent is or comprises eugenol. In some embodiments, the flavoring agent is or comprises thymol. In some embodiments, the flavoring agent is or comprises geraniol. In some embodiments, the flavoring agent is selected from the group consisting of geraniol, (R-) linalool, (R- and/or S-)citronellol, nerol, 8-hydroxylinalool and farnesol.

In some embodiments, the flavoring agent is or comprises vanillin. In some embodiments, the flavoring agent is or comprises ethylvanillin. In some embodiments, the flavoring agent is not vanillin or ethylvanillin.

In some embodiments, the flavor glycoside is selected from the group consisting of 3-methoxystyrene-4-yl-O-β-D-glucopyranoside, 4-ethyl-2-methoxyphenyl- O-β-D-glucopyranoside, phenylmethyl-O-β-D-glucopyranoside, endo-(1S)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-yl-O-β-D-glucopyranoside, 2-methyl-5-(1-methylethyl)phenyl-O-β-D-glucopyranoside, 3,7-dimethyl-6-octen-1-yl-O-β-D-glucopyranoside, 2-methoxy-4-(2-propen-1-yl)phenyl-O-β-D-glucopyranoside, 2-ethyl-4H-pyran-4-one-3-O-β-D-glucopyranoside, 3,7,11-trimethyl-2,6,10-dodecatrien-1-yl-O-β-D-glucopyranoside, 1,3,3-trimethyl-bicyclo[2.2.1]heptan-2-yl-O-β-D-glucopyranoside, 2,5-dimethyl-3(2H)-furanone-4-O-β-D-glucopyranoside, (2E)-3,7-dimethyl-2,6-octadien-1-yl-O-β-D-glucopyranoside, (⅖)-ethyl-5 (or 2)-methyl-3(2H)-furanone-4-O-β-D-glucopyranoside, hex-1-yl-O-β-D-glucopyranoside, 4-[4′-hydroxy phenyl]-butan-2-one-4′-O-β- D- glucopyranoside, (3Z)-3-hexen-1-yl-O-β-D-glucopyranoside, 1-ethenyl-1,5-dimethyl-4-hexen-1-yl-O-β-D-glucopyranoside, methyl-2-aminobenzoate-N- β-D-glucopyranoside, (1R, 2S, 5R)- 5-methyl-2-(1-methylethyl) cyclohexyl-O-β-D-glucopyranoside, 5-ethyl-3-hydroxy-4-methyl-2(5H)-furanone-3-O-β-D-glucopyranoside, 2-methyl-4H-pyran-4-one-3-O-β-D-glucopyranoside, 6,6-dimethylbicyclo(3.1.1)hept-2-ene-2-methyl-O-β-D-glucopyranoside, 5-methyl-3(2H)-furanone-4-O-β-D-glucopyranoside, (2Z)-3,7-dimethyl-2,6-octadien-1-yl-O-β-D-glucopyranoside, 1-octen-3-yl-O-β-D-glucopyranoside, p-mentha-1,8-dien-7-yl-O-β-D-glucopyranoside, 2-phenyleth-1-yl-O-β-D-glucopyranoside, 3,4-(methylenedioxy)phenol, 5-Benzodioxolyl-Oβ-D-glucopyranoside, 4,5-dimethylfuran-2(5H)-one-3-O-β-D-glucopyranoside, (S)-2-(4-methyl-3-cyclohexenyl)-2-propanyl-O-β-D-glucopyranoside, 5-methyl-2-(1-methylethyl)phenyl-O-β-D-glucopyranoside, 3-methoxy-benzyl alcohol-4- O-β- D- glucopyranoside, and combinations thereof.

In some embodiments, the flavor glycoside is selected from the group consisting of 3-methoxystyrene-4-yl-O-β-D-glucopyranoside, 4-ethyl-2-methoxyphenyl- O-β-D-glucopyranoside, phenylmethyl-O-β-D-glucopyranoside, endo-(1S)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-yl-O-β-D-glucopyranoside, 2-methyl-5-(1-methylethyl)phenyl-O-β-D-glucopyranoside, 3,7-dimethyl-6-octen-1-yl-O-β-D-glucopyranoside, 2-methoxy-4-(2-propen-1-yl)phenyl-O-β-D-glucopyranoside, 2-ethyl-4H-pyran-4-one-3-O-β-D-glucopyranoside, 3,7,11-trimethyl-2,6,10-dodecatrien-1-yl-O-β-D-glucopyranoside, 1,3,3-trimethyl-bicyclo[2.2.1]heptan-2-yl-O-β-D-glucopyranoside, 2,5-dimethyl-3(2H)-furanone-4-O-β-D-glucopyranoside, (2E)-3,7-dimethyl-2,6-octadien-1-yl-O-β-D-glucopyranoside, (⅖)-ethyl-5 (or 2)-methyl-3(2H)-furanone-4-O-β-D-glucopyranoside, hex-1-yl-O-β-D-glucopyranoside, 4-[4′-hydroxy phenyl]-butan-2-one-4′-O-β- D- glucopyranoside, (3Z)-3-hexen-1-yl-O-β-D-glucopyranoside, 1-ethenyl-1,5-dimethyl-4-hexen-1-yl-O-β-D-glucopyranoside, methyl-2-aminobenzoate-N- β-D-glucopyranoside, (1R, 2S, 5R)- 5-methyl-2-(1-methylethyl) cyclohexyl-O-β-D-glucopyranoside, 5-ethyl-3-hydroxy-4-methyl-2(5H)-furanone-3-O-β-D-glucopyranoside, 2-methyl-4H-pyran-4-one-3-O-β-D-glucopyranoside, 6,6-dimethylbicyclo(3.1.1)hept-2-ene-2-methyl-O-β-D-glucopyranoside, 5-methyl-3(2H)-furanone-4-O-β-D-glucopyranoside, (2Z)-3,7-dimethyl-2,6-octadien-1-yl-O-β-D-glucopyranoside, 1-octen-3-yl-O-β-D-glucopyranoside, p-mentha-1,8-dien-7-yl-O-β-D-glucopyranoside, 2-phenyleth-1-yl-O-β-D-glucopyranoside, 3,4-(methylenedioxy)phenol, 5-Benzodioxolyl-O-β-D-glucopyranoside, 4,5-dimethylfuran-2(5H)-one-3-O-β-D-glucopyranoside, (S)-2-(4-methyl-3-cyclohexenyl)-2-propanyl-O-β-D-glucopyranoside, 5-methyl-2-(1-methylethyl)phenyl-O-β-D-glucopyranoside, and combinations thereof.

In some embodiments, the flavor glycoside is selected from the group consisting of geranyl β-D- glucoside, (R-)linaloyl β-D-glucoside, (R- and/or S-)citronellyl β-D-glucoside, neryl β-D-glucoside, 8-hydroxylinaloyl glucoside and farnesyl glucoside. In some embodiments, the flavor glycoside is selected from the group consisting of geranyl β-D-glucoside and/or (Rand/or S-)citronellyl β-D-glucoside. In some embodiments, the flavor glycoside is geranyl β-D-glucoside.

In some embodiments, the flavor glycoside is octanyl glycoside, such as octanyl glucoside. In some embodiments, the flavor glycoside is furaneyl glycoside, such as furaneyl glucoside. In some embodiments, the flavor glycoside is hexanyl glycoside, such as hexanyl glucoside.

In some embodiments, the flavor glycoside is not or does not comprise 3-methoxy-benzyl alcohol-4- O-β- D- glucopyranoside.

In some embodiments, the flavor glycoside is present in the oral product in an amount of from about 0.001% to about 20% by weight of the oral product. In some embodiments, the flavor glycoside is present in an amount of from about 0.01% to about 15% by weight of the oral product, such as from about 0.1% to about 10% by weight of the oral product, such as from about 0.5% to about 7.5% by weight of the oral product, such as from about 1% to about 5% by weight of the oral product. In some embodiments, the flavor glycoside is present in an amount of from about 0.1% to about 5% by weight of the oral product, such as from about 0.25% to about 4.5% by weight of the oral product, such as from about 0.5% to about 4% by weight of the oral product, such as from about 0.75% to about 3.5% by weight of the oral product, such as from about 1% to about 3% by weight of the oral product, such as from about 1.5% to about 2.5% by weight of the oral product. In some embodiments, the flavor glycoside is present in an amount of from about 0.001 % to about 2% by weight of the oral product, such as from about 0.001% to about 1% by weight of the oral product. In some embodiments, the flavor glycoside is present in an amount of from about 0.01% to about 1.5% by weight of the oral product, such as from about 0.05% to about 1% by weight of the oral product, such as from about 0.1% to about 0.75% by weight of the oral product, such as from about 0.1% to about 0.5% by weight of the oral product.

The inclusion of a flavoring agent in the form of a flavor glycoside may provide improved levels of control over the release kinetics and release profile of the flavoring agent when the product is placed in the oral cavity of the user. In this regard, it has been found that the inclusion of a flavoring agent in the form of a flavor glycoside may provide slower and more prolonged release of the flavoring agent in use. This prolonged release allows the user to experience the sensorial benefits of the flavoring agent over an extended period of time. This may mean that there is a less pronounced reduction in flavor perceived by the user even after the composition has been in the oral cavity for an extended period of time.

In some embodiments, no more than 50% by weight of the flavoring agent is released from the flavor glycoside within about 5 minutes when placed in the oral cavity of the user. This percentage by weight is based on the total weight of all flavoring agent included in the flavor glycoside. By “no more than X% ... is released within about Y minutes” is meant that, after Y minutes of being held in the oral cavity of the user, the amount of flavoring agent released from the flavor glycoside is no greater than X% by weight. As used herein, the term “released” in the context of the flavoring agent being released from the flavor glycoside refers to the enzymatic cleavage of the flavoring agent from the sugar molecule.

The amount of flavoring agent released from or enzymatically cleaved from the sugar molecule in the flavor glycoside can be measured by any method known to a person skilled in the art. For example, an oral product containing a flavor glycoside as defined herein may be placed under the upper lip of a user. After 30 seconds, the product may be moisturized by using the tip of the tongue and, after 5 minutes, the product may be disgorged into a centrifuge tube and any salivary enzymatic processes stopped by adding methanol (5 mL, p.A.) into the centrifuge tube. The procedure may be carried out at time 0 to act as a control sample (i.e. to measure the initial amount of flavoring agent released), and then repeated at different intervention times (e.g. 5 mins, 10 mins, and the like) as needed in order to measure the amount of flavoring agent released at any one time. The amount of flavoring agent released may then be calculated via measuring the amount of flavor glycoside before being placed in the mouth and then after certain time points.

The centrifuge tubes containing the oral product and methanol (5 mL, p.A.) may then be vortexed for 10 seconds and placed on a plate shaker for 10 minutes to extract the remaining flavor glycoside. Afterwards, the tubes may be centrifuged (13,200 rpm, 10 min) and the supernatant decanted into a safe lock tube (2 mL). The safe lock tube may be centrifuged (13,200 rpm, 10 min) and an aliquot taken for analysis. The amount of flavoring agent released may then be measured using a HPLC system (Knauer, Germany) equipped with a pump (AZURA P2.1S), auto-sampler (AZURA AS 6.1L) and diode-array UV/VIS detector (AZURA DAD 6.1L). Samples may be loaded onto a reversed phase C18 column (Knauer, Eurospher II 100 C18, 3 mm × 100 mm) and separated with a 10 min gradient starting at 90% solvent mixture A (e.g. 0.1% formic acid in water) in solvent mixture B (e.g. 0.1% formic acid in methanol) at a flow rate of 0.5 mL/min changing to 30% A in 5 min and back to 90% A in 0.05 min holding for 4.5 min. The detection UV wavelength may be set at 280 nm. Quantitation may be performed by using an external calibration curve.

In some embodiments, no more than about 40% by weight of the flavoring agent, such as no more than about 35% by weight of the flavoring agent, such as no more than about 30% by weight of the flavoring agent, such as no more than about 25% by weight of the flavoring agent, such as no more than about 20% by weight of the flavoring agent, such as no more than about 15% by weight of the flavoring agent, such as no more than about 10% by weight of the flavoring agent, such as no more than about 5% by weight of the flavoring agent, such as no more than about 1% of the flavoring agent is released from the flavor glycoside within about 5 minutes when placed in the oral cavity of the user. In some embodiments, no more than about 35% by weight of the flavoring agent is released from the flavor glycoside within about 5 minutes when placed in the oral cavity of the user.

In some embodiments, no more than about 50% by weight of the flavoring agent is released from the flavor glycoside within about 10 minutes when placed in the oral cavity. For example, no more than about 40% by weight of the flavoring agent, such as no more than about 30% by weight of the flavoring agent, such as no more than about 25% by weight of the flavoring agent, such as no more than about 20% by weight of the flavoring agent, such as no more than about 15% by weight of the flavoring agent, such as no more than about 10% by weight of the flavoring agent, such as no more than about 5% by weight of the flavoring agent is released from the flavor glycoside within about 10 minutes when placed in the oral cavity of the user.

In some embodiments, no more than about 70% by weight of the flavoring agent is released from the flavor glycoside within about 15 minutes when placed in the oral cavity. For example, no more than about 60% by weight of the flavoring agent, such as no more than about 50% by weight of the flavoring agent, such as no more than about 40% by weight of the flavoring agent, such as no more than about 30% by weight of the flavoring agent, such as no more than about 25% by weight of the flavoring agent, such as no more than about 20% by weight of the flavoring agent, such as no more than about 15% by weight of the flavoring agent, such as no more than about 10% by weight of the flavoring agent, such as no more than about 5% by weight of the flavoring agent is released from the flavor glycoside within about 15 minutes when placed in the oral cavity of the user. In some embodiments, no more than about 60% by weight of the flavoring agent is released from the flavor glycoside within about 15 minutes when placed in the oral cavity of the user.

In some embodiments, no more than about 90% by weight of the flavoring agent is released from the flavor glycoside within about 20 minutes when placed in the oral cavity. In some embodiments, no more than about 80% by weight of the flavoring agent is released from the flavor glycoside within about 20 minutes when placed in the oral cavity. In some embodiments, no more than about 70% by weight of the flavoring agent is released from the flavor glycoside within about 20 minutes when placed in the oral cavity. For example, no more than about 60% by weight of the flavoring agent, such as no more than about 50% by weight of the flavoring agent, such as no more than about 40% by weight of the flavoring agent, such as no more than about 30% by weight of the flavoring agent, such as no more than about 25% by weight of the flavoring agent, such as no more than about 20% by weight of the flavoring agent, such as no more than about 15% by weight of the flavoring agent, such as no more than about 10% by weight of the flavoring agent is released from the flavor glycoside within about 20 minutes when placed in the oral cavity of the user.

In some embodiments, no more than about 90% by weight of the flavoring agent is released from the flavor glycoside within about 30 minutes when placed in the oral cavity. For example, no more than about 85% by weight of the flavoring agent, such as no more than about 80% by weight of the flavoring agent, such as no more than about 75% by weight of the flavoring agent, such as no more than about 70% by weight of the flavoring agent, such as no more than about 60% by weight of the flavoring agent, such as no more than about 50% by weight of the flavoring agent is released from the flavor glycoside within about 30 minutes when placed in the oral cavity of the user.

In some embodiments, no more than about 25% by weight of the flavoring agent is released from the flavor glycoside within about 5 minutes when placed in the oral cavity.

In some embodiments, no more than about 40% by weight of the flavoring agent is released from the flavor glycoside within about 10 minutes when placed in the oral cavity.

In some embodiments, no more than about 65% by weight of the flavoring agent is released from the flavor glycoside within about 20 minutes when placed in the oral cavity.

In some embodiments, from about 0.1 wt% to about 90 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 1 wt% to about 90 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 5 wt% to about 90 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 10 wt% to about 85 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 20 wt% to about 80 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user. As used herein, this means that (in these embodiments), this amount of flavoring agent has been released from the flavor glycoside when the product is tested after 20 minutes of being in the oral cavity of the user. “wt%” in this context refers to the percentage amount of the flavoring agent based on the total amount of the flavoring agent in the flavor glycoside.

In some embodiments, from about 25 wt% to about 75 wt% of the flavoring agent, such as from about 30 wt% to about 70 wt%, such as from about 35 wt% to about 70 wt%, such as from about 40 wt% to about 65 wt%, such as from about 45 wt% to about 60 wt%, such as from about 50 wt% to about 55 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 35 wt% to about 60 wt% of the flavoring agent is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 5 wt% to about 75 wt% of the flavoring agent, such as from about 5 wt% to about 70 wt%, such as from about 5 wt% to about 60 wt%, such as from about 5 wt% to about 50 wt%, such as from about 5 wt% to about 40 wt%, such as from about 5 wt% to about 30 wt% is released from the flavor glycoside about 20 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 0.1 wt% to about 70 wt% of the flavoring agent is released from the flavor glycoside about 15 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 1 wt% to about 65 wt% of the flavoring agent is released from the flavor glycoside about 15 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 5 wt% to about 65 wt% of the flavoring agent is released from the flavor glycoside about 15 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 10 wt% to about 65 wt% of the flavoring agent, such as from about 15 wt% to about 60 wt%, such as from about 20 wt% to about 60 wt%, such as from about 25 wt% to about 55 wt%, such as from about 30 wt% to about 50 wt%, such as from about 35 wt% to about 50 wt%, such as from about 40 wt% to about 45 wt% of the flavoring agent is released from the flavor glycoside about 15 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 25 wt% to about 60 wt% of the flavoring agent is released from the flavor glycoside about 15 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 1 wt% to about 60 wt% of the flavoring agent is released from the flavor glycoside about 15 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 0.01 wt% to about 70 wt% of the flavoring agent is released from the flavor glycoside about 10 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 0.1 wt% to about 70 wt% of the flavoring agent is released from the flavor glycoside about 10 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 1 wt% to about 70 wt% of the flavoring agent is released from the flavor glycoside about 10 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 5 wt% to about 65 wt%, such as from about 10 wt% to about 65 wt% of the flavoring agent, such as from about 15 wt% to about 60 wt%, such as from about 20 wt% to about 55 wt%, such as from about 25 wt% to about 50 wt%, such as from about 30 wt% to about 50 wt%, such as from about 35 wt% to about 45 wt% of the flavoring agent is released from the flavor glycoside about 10 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 20 wt% to about 50 wt% of the flavoring agent is released from the flavor glycoside about 10 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 1 wt% to about 50 wt% of the flavoring agent is released from the flavor glycoside about 10 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 0.01 wt% to about 50 wt% of the flavoring agent is released from the flavor glycoside about 5 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 0.1 wt% to about 50 wt%, such as from about 0.5 wt% to about 50 wt%, such as from about 1 wt% to about 50 wt%, such as from about 5 wt% to about 45 wt% of the flavoring agent, such as from about 10 wt% to about 40 wt%, such as from about 15 wt% to about 35 wt%, such as from about 20 wt% to about 30 wt%, such as from about 25 wt% to about 30 wt% of the flavoring agent is released from the flavor glycoside about 5 minutes after the oral product is placed in the oral cavity of a user. In some embodiments, from about 20 wt% to about 35 wt% of the flavoring agent is released from the flavor glycoside about 5 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, from about 0.1 wt% to about 35 wt% of the flavoring agent is released from the flavor glycoside about 5 minutes after the oral product is placed in the oral cavity of a user.

In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 10 minutes when the oral product is placed in the oral cavity of a user. In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 15 minutes, such as at least about 15 minutes, such as at least about 20 minutes, such as at least about 25 minutes, such as at least about 30 minutes, such as at least about 40 minutes, such as at least about 50 minutes, such as at least about 60 minutes when the oral product is placed in the oral cavity of a user. In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 30 minutes when the oral product is placed in the oral cavity of a user. In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 60 minutes when the oral product is placed in the oral cavity of a user. It has therefore been found that the use of a flavor glycoside in the oral product may prolong the delivery of flavor (from the flavoring agent) to the user during use.

The oral product may comprise an emulsion that comprises a continuous phase and a dispersed phase. The flavor glycoside may be present in the oral product in the continuous or dispersed phase of such an emulsion. For example, the oral product may comprise an emulsion including an oil phase and an aqueous phase, wherein the flavor glycoside is present in the oil phase and/or the aqueous phase. In some embodiments, the flavor glycoside is present in the aqueous phase of such an emulsion. In some embodiments, the oral product comprises an oil-in-water emulsion and the flavor glycoside is present in the aqueous phase of the emulsion.

The emulsion may be any suitable emulsion for inclusion in an oral product. Examples of suitable emulsions that may be included in the product are those described in US 62/945,487, US 62/945,485 and US 62/945,423, the disclosures of which are hereby incorporated by reference. The amount of the emulsion in the oral product may vary and may be any suitable amount for forming a product suitable for oral application. In some embodiments, the emulsion is present in the oral product in an amount of from about 1% to about 75% by weight of the oral product, such as from about 5% to about 60% by weight, such as from about 10% to about 50% by weight, such as from about 15% to about 45% by weight, such as from about 20% to about 40% by weight, such as from about 25% to about 40% by weight, such as from about 30% to about 40% by weight of the oral product.

In some embodiments, the emulsion is in the form of a microemulsion. In some embodiments, the emulsion is in the form of a nanoemulsion. A nanoemulsion is a colloidal particulate system with particulates in the submicron size range. The particulates (referred to herein also as droplets or particles) are generally solid spheres, and the surfaces of such particulates are amorphous and lipophilic with a negative charge. Nanoemulsions generally comprise nanoscale particles or droplets having an average size of less than about 1,000 nm. Nanoemulsions as described herein comprise nanoparticles (or nanodroplets) of the dispersed phase emulsified in the continuous phase. In some embodiments, the nanoemulsion comprises nanoparticles of an oil phase emulsified in water or the aqueous phase. In some embodiments, the oral product comprises a nanoemulsion comprising nanoparticles of an oil phase dispersed in an aqueous phase, wherein the flavor glycoside is contained in the aqueous phase.

Nanoemulsions as described herein generally comprise nanoscale particles having an average size of from about 10 nm to about 1,000 nm, for example, from about 10 nm to about 200 nm, from about 20 nm to about 100 nm, or from about 40 nm to about 100 nm. In some embodiments, the average particle size is about 100 nm, about 90 nm, about 80 nm, about 70 nm, about 60 nm, about 50 nm or about 40 nm. In some embodiments, the average particle size is from about 40 nm to about 80 nm. In some embodiments, the average particle size is from about 40 nm to about 80 nm, and the nanoemulsion is transparent. The size of the nanoparticles may be determined by quasi-electric light scattering (QELS) as described in Bloomfield, Ann. Rev. Biophys. Bioeng., 10:421-450 (1981), incorporated herein by reference. It may also be measured by correlation spectroscopy that analyzes the fluctuation in scattering of light due to Brownian motion, or by transmission electron microscopy (TEM).

Active Ingredient

In some embodiments, the oral product further comprises at least one active ingredient. The active ingredient may be any suitable active ingredient that causes a biological response in a human or animal. The active ingredient as used herein may be a physiologically active material, which is a material intended to achieve or enhance a physiological response. The active substance may for example be selected from nutraceuticals, nootropics, psycho-actives. The active substance may be naturally occurring or synthetically obtained. The active substance may comprise for example nicotine, caffeine, taurine, theine, vitamins such as B6 or B12 or C, melatonin, ginseng, theanine, gamma-aminobutyric acid (GABA), cannabinoids, or constituents, derivatives, or combinations thereof.

In some embodiments, the active ingredient is selected from a nicotine component, a botanical ingredient (e.g., lavender, peppermint, chamomile, basil, rosemary, ginger, ginseng, maca, and tisanes), a stimulant (e.g., caffeine or guarana), an amino acid (e.g., taurine, theanine, phenylalanine, tyrosine, GABA, and tryptophan), a cannabinoid, and/or a pharmaceutical, nutraceutical, or medicinal ingredient (e.g., a vitamin, such as B6, B12, and C).

In some embodiments, the active ingredient comprises nicotine. As described herein, any nicotine stated to be present is taken to be in addition to any nicotine present in any tobacco material (i.e. “additional nicotine”). For example, the oral product may include 1% tobacco material and 2% nicotine, where the nicotine is in addition to any nicotine that may be present in the tobacco material. The terms “tobacco material” and “nicotine” in the context of an active ingredient are therefore not to be taken to be synonymous.

Where present as an active agent, nicotine may be present in any suitable form of nicotine (e.g., free base or salt) for providing oral absorption of at least a portion of the nicotine present. Typically, the nicotine is selected from the group consisting of nicotine free base and a nicotine salt. In some embodiments, nicotine is in its free base form, which can be adsorbed in for example, a microcrystalline cellulose material to form a microcrystalline cellulose-nicotine carrier complex. See, for example, the discussion of nicotine in free base form in U.S. Pat. Pub. No. 2004/0191322 to Hansson, which is incorporated herein by reference.

In some embodiments, at least a portion of the nicotine (which is additional to any tobacco material) can be employed in the form of a salt. Salts of nicotine can be provided using the types of ingredients and techniques set forth in U.S. Pat. No. 2,033,909 to Cox et al. and Perfetti, Beitrage Tabakforschung Int., 12: 43-54 (1983), which are incorporated herein by reference. Further salts are disclosed in, for example, U.S. Pat. No. 9,738,622 to Dull et al., and U.S. Pat. Pub. Nos. 2018/0230126 to Dull et al., 2016/0185750 to Dull et al., and 2018/0051002 to Dull et al., each of which is incorporated herein by reference. Additionally, salts of nicotine are available from sources such as Pfaltz and Bauer, Inc. and K&K Laboratories, Division of ICN Biochemicals, Inc.

In some embodiments, the nicotine (which is additional to any tobacco material) is selected from the group consisting of nicotine free base, a nicotine salt such as hydrochloride, dihydrochloride, monotartrate, bitartrate, sulfate, salicylate, and nicotine zinc chloride.

In some embodiments, at least a portion of the nicotine (which is additional to any tobacco material) can be in the form of a resin complex of nicotine, where nicotine is bound in an ionexchange resin, such as nicotine polacrilex, which is nicotine bound to, for example, a polymethacrilic acid, such as Amberlite IRP64, Purolite C115HMR, or Doshion P551. See, for example, U.S. Pat. No. 3,901,248 to Lichtneckert et al., which is incorporated herein by reference. Another example is a nicotine-polyacrylic carbomer complex, such as with Carbopol 974P. In some embodiments, nicotine may be present in the form of a nicotine polyacrylic complex.

In some embodiments, the nicotine (which is additional to any tobacco material) when present is in a concentration of at least about 0.001 % by weight of the oral product, such as in a range from about 0.001% to about 10%. In some embodiments, the additional nicotine is present in a concentration from about 0.1% to about 10% by weight, such as from about from about 0.1% to about 9%, such as from about 0.2% to about 8%, such as from about 0.3% to about 7%, such as from about 0.4% to about 6%, such as from about 0.5% to about 5%, such as from about 0.6% to about 4%, such as from about 0.7% to about 3%, such as from about 0.8% to about 2%, or from about 0.9% to about 1%, calculated as the free base and based on the total weight of the oral product. In some embodiments, the nicotine component is present in a concentration from about 0.1% to about 3% by weight, such as from about from about 0.1% to about 2.5%, such as from about 0.1% to about 2.0%, such as from about 0.1% to about 1.5%, such as from about 0.1% to about 1% by weight, calculated as the free base and based on the total weight of the oral product.

It is noted that these above ranges also apply to each of the other additional active ingredients noted herein.

In some embodiments, the active ingredient comprises caffeine, melatonin or vitamin B12. In some embodiments, the active ingredient comprises caffeine.

In some embodiments, the active ingredient comprises a cannabinoid. The cannabinoid may be a derivative or extract of cannabis. Cannabinoids are a class of natural or synthetic chemical compounds which act on cannabinoid receptors (i.e., CB1 and CB2) in cells that repress neurotransmitter release in the brain. Cannabinoids are cyclic molecules exhibiting particular properties such as the ability to easily cross the blood-brain barrier. Cannabinoids may be naturally occurring (Phytocannabinoids) from plants such as cannabis, (endocannabinoids) from animals, or artificially manufactured (synthetic cannabinoids). Cannabis species express at least 85 different phytocannabinoids, and these may be divided into subclasses, including cannabigerols, cannabichromenes, cannabidiols, tetrahydrocannabinols, cannabinols and cannabinodiols, and other cannabinoids, such as cannabigerol (CBG), cannabichromene (CBC), cannabidiol (CBD), tetrahydrocannabinol (THC), cannabinol (CBN) and cannabinodiol (CBDL), cannabicyclol (CBL), cannabivarin (CBV), tetrahydrocannabivarin (THCV), cannabidivarin (CBDV), cannabichromevarin (CBCV), cannabigerovarin (CBGV), cannabigerol monomethyl ether (CBGM), cannabinerolic acid, cannabidiolic acid (CBDA), Cannabinol propyl variant (CBNV), cannabitriol (CBO), tetrahydrocannabmolic acid (THCA), and tetrahydrocannabivarinic acid (THCV A).

In some embodiments, the cannabinoid is selected from the group consisting of cannabigerol (CBG), cannabichromene (CBC), cannabidiol (CBD), tetrahydrocannabinol (THC), cannabinol (CBN) and cannabinodiol (CBDL), cannabicyclol (CBL), cannabivarin (CBV), tetrahydrocannabivarin (THCV), cannabidivarin (CBDV), cannabichromevarin (CBCV), cannabigerovarin (CBGV), cannabigerol monomethyl ether (CBGM), cannabinerolic acid, cannabidiolic acid (CBDA), Cannabinol propyl variant (CBNV), cannabitriol (CBO), tetrahydrocannabmolic acid (THCA), tetrahydrocannabivarinic acid (THCV A), and mixtures thereof. In some embodiments, the cannabinoid comprises at least tetrahydrocannabinol (THC). In some embodiments, the cannabinoid is tetrahydrocannabinol (THC). In some embodiments, the cannabinoid comprises at least cannabidiol (CBD). In some embodiments, the cannabinoid is cannabidiol (CBD).

In some embodiments, the cannabinoid is cannabidiol (CBD) or a pharmaceutically acceptable salt thereof. In some embodiments, the cannabidiol is synthetic cannabidiol. In some embodiments, the cannabinoid is added to the composition in the form of an isolate. In some embodiments, the cannabidiol is added to the composition in the form of an isolate. An isolate is an extract from a plant, such as cannabis, where the active material of interest (in this case the cannabinoid, such as CBD) is present in a high degree of purity, for example greater than 95%, greater than 96%, greater than 97%, greater than 98%, or around 99% purity.

In some embodiments, the cannabinoid is an isolate of CBD in a high degree of purity, and the amount of any other cannabinoid in the composition is no greater than about 1% by weight of the oral product, such as no greater than about 0.5% by weight of the oral product, such as no greater than about 0.1% by weight of the oral product, such as no greater than about 0.01% by weight of the oral product.

The choice of cannabinoid and the particular percentages thereof which may be present within the disclosed product will vary depending upon the desired flavor, texture, and other characteristics of the oral product.

In some embodiments, the cannabinoid (such as cannabidiol) is present in the oral product in a concentration of at least about 0.001 % by weight of the oral product, such as in a range from about 0.001% to about 20% by weight of the oral product. In some embodiments, the cannabinoid (such as cannabidiol) is present in the oral product in a concentration of from about 0.1% to about 15% by weight, based on the total weight of the oral product. In some embodiments, the cannabinoid (such as cannabidiol) is present in a concentration from about 1% to about 15% by weight, such as from about from about 5% to about 15% by weight, based on the total weight of the oral product. In some embodiments, the cannabinoid (such as cannabidiol) is present in the oral product in a concentration of from about 0.5% to about 10% by weight, such as from about 1% to about 7.5% by weight, such as from 1.5% to about 5% by weight, such as from about 1.5% to about 2.5% by weight, based on the total weight of the oral product.

As described hereinabove, the oral product may comprise an emulsion that comprises a continuous phase and a dispersed phase. In some embodiments, the emulsion comprises an aqueous phase and a lipophilic phase, wherein the at least one flavor glycoside is present in the aqueous phase and the at least one active ingredient is present in the lipophilic phase. In some embodiments, the oral product comprises an oil-in-water emulsion, wherein the at least one flavor glycoside is present in the aqueous phase and the at least one active ingredient is present in the oil phase. The emulsion may be an oil-in-water nanoemulsion, wherein the at least one active ingredient is held in the oil nanodroplets present within the nanoemulsion.

Additives

In some embodiments, the oral product further comprises one or more additional flavoring agents. As used herein, the term “additional flavoring agent” refers to a flavoring agent that is included in addition to the flavoring agent in the flavor glycoside. The additional flavoring agent may be present in its standard form; i.e. without having been bound to a sugar molecule via a glycosidic bond. As such, the composition may comprise a flavor glycoside in addition to a flavoring agent that is not part of a flavor glycoside.

In some embodiments, the additional flavoring agent is different from the flavoring agent in the flavor glycoside. For example, in embodiments in which the flavor glycoside comprises a raspberry ketone bound to a sugar molecule via a glycosidic bond, the additional flavoring agent may be any flavoring agent other than a raspberry ketone.

In some embodiments, the additional flavoring agent is the same as the flavoring agent in the flavor glycoside. For example, in embodiments in which the flavor glycoside comprises a raspberry ketone bound to a sugar molecule via a glycosidic bond, the additional flavoring agent may be a raspberry ketone that is not bound to a sugar molecule via a glycosidic bond.

In some embodiments, there may be two or more additional flavoring agents. The two or more additional flavoring agents may both be different from the flavoring agent in the flavor glycoside, or one of the additional flavoring agents may be the same as the flavoring agent in the flavor glycoside whilst the other(s) may be different from the flavoring agent in the flavor glycoside.

The additional flavoring agent(s) may be selected from any of the flavoring agents described hereinabove in respect of suitable flavoring agents for the flavor glycoside. For conciseness, these are not repeated here, but the same disclosure as hereinabove equally applies for the one or more additional flavoring agents.

In some embodiments, the rate of release of the flavoring agent in the flavor glycoside is slower than the rate of release of the one or more additional flavoring agents, wherein the rate of release is measured as the rate at which a flavoring agent is released into a user’s mouth when the oral product is placed in the oral cavity of the user. For example, when the product is placed in the oral cavity of the user, the user may experience the taste of the additional flavoring agent before experiencing the taste of the flavoring agent that is cleaved from the sugar molecule in the flavor glycoside.

In some embodiments, the one or more additional flavoring agents is released into the oral cavity of a user during use prior to the release of the flavoring agent from the flavor glycoside.

For example, in some embodiments, the rate of release of the one or more additional flavoring agents may be from about 1.1 to about 20 times faster than the rate of release of the flavoring agent from the flavor glycoside, such as from about 1.2 to about 15 times faster, such as from about 1.3 to about 10 times faster, such as from about 1.4 to about 7.5 times faster, such as from about 1.5 to about 5 times faster. In some embodiments, the additional flavoring agent may be released into the user’s cavity (i.e. perceived by the user) within a period of from about 1 second to about 20 minutes, such as from about 5 seconds to about 15 minutes, such as from about 10 seconds to about 10 minutes, such as from about 30 seconds to about 5 minutes, as compared with the rate of release of the flavoring agent from the flavor glycoside which is as described hereinabove.

Where the additional flavoring agent(s) is different from the flavoring agent in the flavor glycoside, such distinction in the rates of the release of the flavoring agents may provide the user with a flavor profile that changes over a period of time; i.e. the flavor released from the product may change over a period of time as the flavoring agent in the flavor glycoside is released after the additional flavoring agent(s).

Where the additional flavoring agent(s) is the same as the flavoring agent in the flavor glycoside, such distinction in the rates of the release of the flavoring agents may further prolong the period of time for which the flavor is delivered to the user. For example, flavor may be delivered to the user rapidly (i.e. within a period of from about 1 second to about 10 minutes) from the additional flavoring agent(s), and then the flavoring agent from the flavor glycoside may subsequently be released to prolong the delivery of the flavor to the user. In some embodiments, the flavor is delivered to the user for a period of at least about 10 minutes, such as at least about 15 minutes, such as at least about 20 minutes, such as at least about 25 minutes, such as at least about 30 minutes, such as at least about 40 minutes, such as at least about 50 minutes, such as at least about 60 minutes when the oral product is placed in the oral cavity of a user.

The amount of any additional flavoring agent included in the oral product can vary. In some embodiments, the oral product comprises one or more additional flavoring agents in an amount of up to about 10% by weight, such as up to about 5% by weight, such as up to about 1% by weight of the oral product. In some embodiments, the oral product comprises one or more additional flavoring agents in an amount of from about 0.01% to about 10% by weight, such as from about 0.1% to about 5% by weight, such as from about 0.5% to about 1% by weight of the oral product.

Depending on the type of oral product being processed, the product may include one or more additional components in addition to the filler or bulking agent and flavor glycoside as described above. For example, the oral product may further comprise an additive selected from the group consisting of binders, pH adjusting agents, buffering agents, salts, sweeteners, colorants, oral care additives, disintegration aids, antioxidants, humectants, preservatives, or mixtures thereof. Such additives may be included in addition to the at least one filler or bulking agent as described hereinabove.

In some embodiments, the oral product may further comprise at least one binder. A binder (or combination of binders) may be employed in the composition in certain embodiments, in amounts sufficient to provide the desired physical attributes and physical integrity to the composition. Binders can be organic or inorganic, or a combination thereof. Representative binders include cellulose derivatives, povidone, sodium alginate, starch-based binders, pectin, carrageenan, pullulan, zein, and the like, and combinations thereof. The amount of binder utilized in the product can vary, but may be up to about 30% by weight, and certain embodiments are characterized by a binder content of at least about 0.1% by weight, such as from about 1% to about 30% by weight, or about 1% to about 10% by weight, based on the total weight of the oral product. In some embodiments, the binder comprises a cellulose derivative. In certain embodiments, the cellulose derivative is a cellulose ether (including carboxyalkyl ethers), meaning a cellulose polymer with the hydrogen of one or more hydroxyl groups in the cellulose structure replaced with an alkyl, hydroxyalkyl, or aryl group. Non-limiting examples of such cellulose derivatives include methylcellulose, hydroxypropylcellulose (“HPC”), hydroxypropylmethylcellulose (“HPMC”), hydroxyethyl cellulose, and carboxymethylcellulose (“CMC”). In some embodiments, the cellulose derivative is HPC. In some embodiments, the cellulose derivative is a combination of HPC and HPMC. In some embodiments, the oral product comprises from about 1% to about 10% of the cellulose derivative (such as HPC) by weight of the oral product, with certain embodiments comprising from about 1% to about 5% by weight of cellulose derivative (such as HPC), based on the weight of the product.

In certain embodiments, the binder includes a gum, for example, a natural gum. As used herein, a natural gum refers to polysaccharide materials of natural origin that have binding properties, and which are also useful as a thickening or gelling agents. Representative natural gums derived from plants, which are typically water soluble to some degree, include xanthan gum, guar gum, gum arabic, ghatti gum, gum tragacanth, karaya gum, locust bean gum, gellan gum, and combinations thereof. When present, natural gum binder materials may be present in an amount of up to about 5% by weight, for example, from about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, or about 1%, to about 2, about 3, about 4, or about 5% by weight, based on the total weight of the product.

In some embodiments, the oral product further comprises a taste modifying agent. A taste modifying agent may mask the bitterness of any tobacco and/or any additional active or flavoring ingredient in the product. The taste modifying agent may improve the organoleptic properties of an oral product as disclosed herein, and may serve to mask, alter, block, or improve e.g., the flavor of a product as described herein. Non-limiting examples of such taste modifiers include analgesic or anesthetic herbs, spices, and flavors which produce a perceived cooling (e.g., menthol, eucalyptus, mint), warming (e.g., cinnamon), or painful (e.g., capsaicin) sensation. Certain taste modifiers fall into more than one overlapping category. In some embodiments, the taste modifier is capsaicin. In some embodiments, the taste modifier is adenosine monophosphate (AMP). AMP is a naturally occurring nucleotide substance which can block bitter food flavors or enhance sweetness. It does not directly alter the bitter flavor, but may alter human perception of “bitter” by blocking the associated receptor. In some embodiments, the taste modifier is lactisole. Lactisole is an antagonist of sweet taste receptors. Temporarily blocking sweetness receptors may accentuate e.g., savory notes. When present, a representative amount of taste modifying agent may be about 0.01% by weight or more, about 0.1% by weight or more, or about 1.0% by weight or more, but may make up less than about 10% by weight of the total weight of the product, (e.g., from about 0.01%, about 0.05%, about 0.1%, or about 0.5%, to about 1%, about 5%, or about 10% by weight of the total weight of the product).

In some embodiments, the oral product comprises at least one humectant. Examples of suitable humectants that may be included in the composition include, but are not limited to, glycerin, 1,2-propanediol (propylene glycol), 1,3-propanediol, dipropylene glycol, sorbitol, xylitol, mannitol, and the like. In some embodiments, the humectant is or comprises glycerin. In some embodiments, the oral product comprises glycerin. In some embodiments, the humectant is or comprises propylene glycol. In some embodiments, the oral product comprises propylene glycol. The amount of humectant utilized in the oral product can vary, but may be up to about 5% by weight, and certain embodiments can be characterized by a humectant content of at least about 1% by weight, such as about 2 to about 5% by weight of the oral product. In some embodiments, the humectant (such as glycerin and/or propylene glycol) may be present in an amount of from about 0.01% to about 25% by weight of the oral product, such as from about 0.1% to about 20% by weight of the oral product, such as from about 0.5% to about 15% by weight of the oral product, such as from about 1% to about 10% by weight of the oral product, such as from about 5% to about 10% by weight of the oral product.

In order to improve the sensory properties of the oral product, one or more sweeteners may be added. The sweeteners can be any sweetener or combination of sweeteners, in natural or artificial form, or as a combination of natural and artificial sweeteners. Examples of natural sweeteners include fructose, sucrose, glucose, maltose, mannose, galactose, lactose, stevia, honey, and the like. Examples of artificial sweeteners include sucralose, isomaltulose, maltodextrin, saccharin, aspartame, acesulfame K, neotame and the like. In some embodiments, the sweetener comprises one or more sugar alcohols. Sugar alcohols are polyols derived from monosaccharides or disaccharides that have a partially or fully hydrogenated form. Sugar alcohols have, for example, about 4 to about 20 carbon atoms and include erythritol, arabitol, ribitol, isomalt, maltitol, dulcitol, iditol, mannitol, xylitol, lactitol, sorbitol, and combinations thereof (e.g., hydrogenated starch hydrolysates). In some embodiments, the sweetener is selected from the group consisting of fructose, sucrose, glucose, maltose, mannose, galactose, lactose, stevia, honey, sucralose, isomaltulose, maltodextrin, saccharin, aspartame, acesulfame K, neotame, erythritol, arabitol, ribitol, isomalt, maltitol, dulcitol, iditol, mannitol, xylitol, lactitol, sorbitol, and mixtures thereof. In some embodiments, the sweetener is selected from the group consisting of sucralose, acesulfame K, aspartame, maltodextrin, mannitol, sucrose, and mixtures thereof. In some embodiments, the sweetener may be sucralose and/or acesulfame K. When present in the oral product, the sweetener (such as sucralose and/or acesulfame K) may be present in an amount of from about 0.001 % to about 5% by weight, such as from about 0.01% to about 3% by weight, such as from about 0.1% to about 1% by weight of the oral product.

Non-limiting examples of suitable buffering agents that may be included in the oral product include alkali metals acetates, glycinates, phosphates, glycerophosphates, citrates, carbonates, hydrogen carbonates, borates, or mixtures thereof. In some embodiments in which a buffering agent is present, the buffering agent is selected from the group consisting of sodium carbonate, sodium bicarbonate, sodium phosphate, ammonium phosphate, and mixtures thereof. In some embodiments, the buffering agent is sodium bicarbonate and/or sodium carbonate. Where present, the buffering agent (e.g. sodium bicarbonate and/or sodium carbonate) may be included in an amount less than about 5% based on the weight of the oral product; for example, from about 0.5% to about 5%, such as, e.g., from about 0.75% to about 4%, from about 0.75% to about 3%, or from about 1% to about 2% by weight, based on the total weight of the oral product.

The oral product may further comprise a salt. This may be included in an amount sufficient to provide desired sensory attributes to the product. Non-limiting examples of suitable salts include sodium chloride, potassium chloride, ammonium chloride, flour salt, sodium acetate, sodium citrate, and the like. The salt may be included in any suitable amount, such as at least about 0.5% by weight, such as at least about 1% by weight, such as at least about 1.5% by weight of the composition. In some embodiments, the oral product may comprise salt in an amount of from about 0.5% to about 10% by weight, such as from about 1% to about 7.5% by weight, such as from about 1.5% to about 5% by weight, based on the total weight of the oral product.

Other ingredients such as preservatives (e.g., potassium sorbate), disintegration aids (e.g., croscarmellose sodium, crospovidone, sodium starch glycolate, pregelatinized corn starch, and the like), and/or antioxidants can also be used. Typically, such ingredients, where used, are used in amounts of up to about 10% by weight, for example at least about 0.1% by weight, such as about 0.5 to about 10% by weight of the oral product. A disintegration aid may be employed in an amount sufficient to provide control of desired physical attributes of the oral product such as, for example, by providing loss of physical integrity and dispersion of the various component materials upon contact of the formulation with water (e.g., by undergoing swelling upon contact with water).

In some embodiments (for example, where the product comprises an emulsion), the composition comprises one or more emulsifying agents. By “emulsifying agent” is meant a substance which aids in the formation and stabilization of emulsions by promoting dispersion of hydrophobic and hydrophilic (e.g., oil and water) components. In general, emulsifying agents are amphiphilic molecules chosen from, for example, nonionic and ionic amphiphilic molecules. The expression “amphiphilic molecule” means any molecule of bipolar structure comprising at least one hydrophobic portion and at least one hydrophilic portion and having the property of reducing the surface tension of water and of reducing the interface tension between water and an oily phase. Emulsifying agents/amphiphilic molecules as provided herein are also referred to as, for example, surfactants and emulsifiers.

In some embodiments, the emulsifying agent is selected from the group consisting of small molecule surfactants, phospholipids, proteins, polysaccharides, and mixtures thereof. The emulsifying agent may be selected from any suitable emulsifying agent, such as those described in detail in US 62/945,487, US 62/945,485 and US 62/945,423, which are hereby incorporated by reference. In some embodiments, the one or more emulsifying agents is selected from the group consisting of glycol distearate, sorbitan trioleate, sorbitan tristearate, sorbitan triisostearate, glyceryl isostearate, propylene glycol isostearate, glycol stearate, sorbitan sesquioleate, glyceryl stearate, lecithin, sorbitan oleate, sorbitan monostearate, sorbitan stearate, sorbitan isostearate, steareth-2, oleth-2, PEG-7 hydrogenated castor oil, laureth-2, sorbitan palmitate, laureth-3, glyceryl laurate, ceteth-2, PEG-30 dipolyhdroxystearate, glyceryl stearate SE, sorbitan stearate (and) sucrose cocoate, PEG-4 dilaurate, methyl glucose sesquistearate, PEG-8 dioleate, sorbitan laurate, PEG-40 sorbitan peroleate, laureth-4, PEG-7 glyceryl cocoate, PEG-20 almond glycerides, PEG-25 hydrogenated castor oil, stearamide MEA, glyceryl stearate (and) PEG-100 stearate, polysorbate 81, polysorbate 85, polysorbate 65, PEG-7 glyceryl cocoate, PEG-8 stearate, PEG-8 caprate, PEG-35 almond glycerides, PEG-6 laurate, laureth-7, steareth-10, isotrideceth-8, PEG-35 castor oil, isotrideceth-9, PEG-40 castor oil, ceteareth-12, laureth-9, PEG-40 hydrogenated castor oil, PEG-20 glyceryl isostearate, PEG-20 stearate, PEG-40 sorbitan perisostearate, PEG-7 olivate, cetearyl glucoside, PEG-8 oleate, polyglyceryl-3 methylglucose distearate, oleth-10, oleth-10/polyoxyl 10 oleyl ether NF, ceteth-10, PEG-8 laurate, cocamide MEA, polysorbate 60, polysorbate 80, isosteareth-20, PEG-60 almond glycerides, PEG-20 methyl glucose sesquistearate, ceteareth-20, oleth-20, steareth-20, steareth-21, ceteth-20, isoceth-20, polysorbate 20, polysorbate 40, ceteareth-25, ceteareth-30, PEG-30 stearate, laureth-23, PEG-75 lanolin, polysorbate 20, PEG-40 stearate, PEG-100 stearate, steareth-100, PEG-80 sorbitan laurate, polyoxyethylene stearate (e.g. polyoxyethylene (40) stearate), polyoxyethylene ether, and mixtures thereof. In some embodiments, the emulsifying agent is or comprises lecithin. In some embodiments, the emulsifying agent is or comprises polyoxyethylene (40) stearate.

In some embodiments, the one or more emulsifying agents have an overall HLB value in the range of from about 10 to about 15, such as from about 11 to about 15, such as from about 11 to about 14, such as from about 11 to about 13.5. As will be understood by one skilled in the art, HLB is the hydrophilic-lipophilic balance of an emulsifying agent or surfactant is a measure of the degree to which it is hydrophilic or lipophilic. The HLB value may be determined by calculating values for the different regions of the molecule, as described by Griffin in Griffin, William C. (1949), “Classification of Surface-Active Agents by ‘HLB’” (PDF), Journal of the Society of Cosmetic Chemists, 1 (5): 311-26 and Griffin, William C. (1954), “Calculation of HLB Values of Non-Ionic Surfactants” (PDF), Journal of the Society of Cosmetic Chemists, 5 (4): 249-56, and by Davies in Davies JT (1957), “A quantitative kinetic theory of emulsion type, I. Physical chemistry of the emulsifying agent” (PDF), Gas/Liquid and Liquid/Liquid Interface, Proceedings of the International Congress of Surface Activity, pp. 426-38. HLB value may be determined in accordance with the industry standard text book, namely “The HLB SYSTEM, a time-saving guide to emulsifier selection” ICI Americas Inc., Published 1976 and Revised, March, 1980. The HLB values of the emulsifiers described herein were determined in accordance with this standard method.

In some embodiments, the oral product comprises at least two emulsifying agents which have different HLB values. In some embodiments, the oral product comprises a first emulsifying agent with a low HLB value, and a second emulsifying agent with a high HLB value. In some embodiments, the oral product comprises a first emulsifying agent having an HLB value of from about 1 to about 9 and a second emulsifying agent having an HLB value of from about 10 to about 20. In some embodiments, the first emulsifying agent is or comprises lecithin. In some embodiments, the first emulsifying agent is or comprises soy lecithin. In some embodiments, the second emulsifying agent is or comprises polyoxyethylene stearate (e.g. polyoxyethylene (40) stearate). In some embodiments, the emulsifying agent is or comprises a combination of lecithin (e.g. soy lecithin) and polyoxyethylene stearate (e.g. polyoxyethylene (40) stearate).

In some embodiments, the one or more emulsifying agents may be present in an emulsion in an amount of from about 0.1% to about 20% by weight of the oral product, such as from about 1% to about 15% by weight, such as from about 2.5% to about 10% by weight, such as from about 5% to about 10% by weight of the oral product. In some embodiments, the emulsion comprises a combination of lecithin and Myrj 52 (polyoxyethylene (40) stearate) in an amount of from about 0.1% to about 20% by weight of the oral product, such as from about 1% to about 15% by weight, such as from about 2.5% to about 10% by weight, such as from about 5% to about 10% by weight of the oral product.

Total Composition & Pouched Product

The compositions as described herein are configured for oral use. The term “configured for oral use” as used herein means that the product is provided in a form such that during use, saliva in the mouth of the user causes one or more of the components of the composition (e.g., flavoring agents and/or active ingredients) to pass into the mouth of the user. In certain embodiments, the composition is adapted to deliver components to a user through mucous membranes in the user’s mouth, the user’s digestive system, or both, and, in some instances, said component is an active ingredient and/or flavoring agent that can be absorbed through the mucous membranes in the mouth or absorbed through the digestive tract when the product is used.

In some embodiments, the oral product is in liquid form. In some embodiments, the oral product is in the form of a gel.

In some embodiments, the oral product is in the form of a solid. As used herein, the term “solid” means that the compositions can substantially sustain their physical shape when unsupported by external means, e.g. packaging etc. Thus, they are considered to be solid, solid like, in solid form or in solid-like form at room temperature. For the avoidance of doubt the solid product remains substantially solid at up to 30° C. In some embodiments, the oral product is in solid form, such as in the form of loose moist snuff, loose dry snuff, chewing tobacco-type form, pelletized pieces, extruded or formed strips, pieces, rods, or sticks, finely divided ground powders, finely divided or milled agglomerates of powdered pieces and components, flake-like pieces, molded processed pieces, pieces of gum, rolls of tape-like films, readily water-dissolvable or water-dispersible films or strips, or capsule-like materials. In some embodiments, the oral product may be a chewing gum product. In some embodiments, the oral product is in the form of chewing tobacco. In some embodiments, the oral product is in the form of moist snuff or snus, which may or may not contain tobacco.

Certain products configured for oral use are in the form of pastilles. As used herein, the term “pastille” refers to a dissolvable oral product made by solidifying a liquid or gel composition so that the final product is a somewhat hardened solid gel. The rigidity of the gel is highly variable. Certain products can exhibit, for example, one or more of the following characteristics: crispy, granular, chewy, syrupy, pasty, fluffy, smooth, and/or creamy. In certain embodiments, the desired textural property can be selected from the group consisting of adhesiveness, cohesiveness, density, dryness, fracturability, graininess, gumminess, hardness, heaviness, moisture absorption, moisture release, mouthcoating, roughness, slipperiness, smoothness, viscosity, wetness, and combinations thereof.

The oral product of the present disclosure may be dissolvable. As used herein, the terms “dissolve,” “dissolving,” and “dissolvable” refer to compositions having aqueous-soluble components that interact with moisture in the oral cavity and enter into solution, thereby causing gradual consumption of the product. According to one aspect, the dissolvable product is capable of lasting in the user’s mouth for a given period of time until it completely dissolves. Dissolution rates can vary over a wide range, from about 1 minute or less to about 60 minutes. For example, fast release compositions typically dissolve and/or release the active substance in about 2 minutes or less, often about 1 minute or less (e.g., about 50 seconds or less, about 40 seconds or less, about 30 seconds or less, or about 20 seconds or less). Dissolution can occur by any means, such as melting, mechanical disruption (e.g., chewing), enzymatic or other chemical degradation, or by disruption of the interaction between the components of the composition. In some embodiments, the product can be meltable as discussed, for example, in U.S. Pat. App. Pub. No. 2012/0037175 to Cantrell et al. In other embodiments, the products do not dissolve during the product’s residence in the user’s mouth.

In some embodiments, the oral product may be in the form of a powder. The powder may be a free-flowing powder. The powder may be contained in loose form within a container, and may thus be used in a form similar to tobacco snuff where the user takes a pinch of powder from the container and places the powder in the oral cavity. Alternatively or additionally, the powder may be incorporated into a moisture-permeable (e.g. saliva-permeable) pouch, similar to a snus-type product. The pouched product may be configured for insertion into the oral cavity of a user; i.e. it may be a pouched oral product.

In some embodiments, the product of the present disclosure is in the form of a pouched oral product. Such a pouched product comprises the oral product as described herein, disposed within a moisture-permeable container (e.g., a water-permeable pouch or saliva-permeable pouch). For example, the pouched product may comprise the oral product in a powder form incorporated within the saliva-permeable pouch.

Therefore, according to some embodiments described herein, there is provided a pouched oral product comprising a saliva permeable pouch and an oral product incorporated within the pouch, wherein the oral product comprises (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond, wherein the oral product comprises no more than about 5% by weight of a tobacco material, excluding any nicotine present, based on the total weight of the product; and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes. The oral product incorporated within the pouch may be in the form of a powder, for example.

Such compositions in the moisture-permeable pouch format are typically used by placing one pouch containing the composition in the mouth of a human subject/user. Generally, the pouch is placed somewhere in the oral cavity of the user, for example under the lips, in the same way as moist snuff products are generally used. The pouch preferably is not chewed or swallowed. Exposure to saliva then causes some of the components of the composition therein (e.g., flavoring agents and/or active ingredients) to pass through e.g., the moisture-permeable pouch and provide the user with flavor and satisfaction, and the user is not required to spit out any portion of the composition. After about 10 minutes to about 60 minutes, typically about 15 minutes to about 45 minutes, of use/enjoyment, substantial amounts of the composition have been ingested by the human subject, and the pouch may be removed from the mouth of the human subject for disposal.

Certain embodiments of the disclosure will be described with reference to FIG. 2 of the accompanying drawing, and these described embodiments involve snus-type products having an outer pouch and containing a composition as described herein. As explained in greater detail below, such embodiments are provided by way of example only, and the pouched products of the present disclosure can include the composition in other forms. The composition/construction of such packets or pouches, such as the container pouch 102 in the embodiment illustrated in FIG. 2, may be varied. Referring to FIG. 2, there is shown a first embodiment of a pouched product 100. The pouched product 100 includes a moisture-permeable container in the form of a pouch 102, which contains an oral product 104 comprising at least one tobacco material and at least one flavor glycoside as described herein.

In some embodiments, the pouch is saliva-permeable. This means that the pouch is made of a saliva-permeable pouch material. In some embodiments, the pouch material is a fleece material. In some embodiments, the pouch material is a non-woven material. In some embodiments, the pouch material is a non-woven fleece material. In some embodiments, the pouch material comprises viscose, such as viscose rayon fibers. In some embodiments, the pouch material comprises regenerated cellulose fibers. In some embodiments, the pouch material comprises polyester fibers; the polyester fibers may constitute the pouch material or may be included in combination with viscose (such as regenerated cellulose fibers).

In some embodiments, the pouch material comprises a binder that provides for heat sealing of the pouches during manufacture. In some embodiments, the pouch material comprises an acrylic binder. In some embodiments, the pouch material comprises an acrylic binder in combination with viscose and/or polyester fibers.

Suitable packets, pouches or containers of the type used for the manufacture of smokeless tobacco products are available under the tradenames CatchDry, Ettan, General, Granit, Goteborgs Rape, Grovsnus White, Metropol Kaktus, Mocca Anis, Mocca Mint, Mocca Wintergreen, Kicks, Probe, Prince, Skruf and TreAnkrare. The composition may be contained in pouches and packaged, in a manner and using the types of components used for the manufacture of conventional snus types of products. The pouch provides a moisture-permeable container of a type that may be considered to be similar in character to the mesh-like type of material that is used for the construction of a tea bag. Components of the composition readily diffuse through the pouch and into the mouth of the user.

Non-limiting examples of suitable types of pouches are set forth in, for example, U.S. Pat. Nos. 5,167,244 to Kjerstad and 8,931,493 to Sebastian et al.; as well as U.S. Pat. App. Pub. Nos. 2016/0000140 to Sebastian et al.; 2016/0073689 to Sebastian et al.; 2016/0157515 to Chapman et al.; and 2016/0192703 to Sebastian et al., each of which is incorporated herein by reference. Pouches can be provided as individual pouches, or a plurality of pouches (e.g., 2, 4, 5, 10, 12, 15, 20, 25 or 30 pouches) can be connected or linked together (e.g., in an end-to-end manner) such that a single pouch or individual portion can be readily removed for use from a one-piece strand or matrix of pouches. The pouch may be formed of a moisture-permeable non-woven fabric, such as viscose for example.

An example pouch may be manufactured from materials, and in such a manner, such that during use by the user, the pouch undergoes a controlled dispersion or dissolution. Such pouch materials may have the form of a mesh, screen, perforated paper, permeable fabric, or the like. For example, pouch material manufactured from a mesh-like form of rice paper, or perforated rice paper, may dissolve in the mouth of the user. As a result, the pouch and composition each may undergo complete dispersion within the mouth of the user during normal conditions of use, and hence the pouch and composition both may be ingested by the user. Other examples of pouch materials may be manufactured using water dispersible film forming materials (e.g., binding agents such as alginates, carboxymethylcellulose, xanthan gum, pullulan, and the like), as well as those materials in combination with materials such as ground cellulosics (e.g., fine particle size wood pulp). Preferred pouch materials, though water dispersible or dissolvable, may be designed and manufactured such that under conditions of normal use, a significant amount of the composition contents permeate through the pouch material prior to the time that the pouch undergoes loss of its physical integrity. If desired, flavoring ingredients, disintegration aids, and other desired components, may be incorporated within, or applied to, the pouch material.

The amount of the oral product contained within each pouched product unit, for example, a pouch, may vary. In some embodiments, the weight of the composition within each pouch is at least about 50 mg, for example, from about 50 mg to about 1 gram (1,000 mg), such as from about 100 mg to about 900 mg, such as from about 200 mg to about 800 mg, such as from about 500 mg to about 700 mg. In some smaller embodiments, the weight of the composition within each pouch may be from about 100 mg to about 300 mg. For a larger embodiment, the weight of the composition within each pouch may be from about 300 mg to about 700 mg. If desired, other components can be contained within each pouch. For example, at least one flavored strip, piece or sheet of flavored water dispersible or water soluble material (e.g., a breath-freshening edible film type of material) may be disposed within each pouch along with or without at least one capsule. Such strips or sheets may be folded or crumpled in order to be readily incorporated within the pouch. See, for example, the types of materials and technologies set forth in U.S. Pat. Nos. 6,887,307 to Scott et al. and 6,923,981 to Leung et al.; and The EFSA Journal (2004) 85, 1-32; which are incorporated herein by reference.

In some embodiments, the at least one flavor glycoside is positioned at a first position on an inner surface of the saliva permeable pouch. Alternatively or in addition, the at least one flavor glycoside may be positioned on an external surface of the saliva permeable pouch. Alternatively, the at least one flavor glycoside may be incorporated within the oral product that is included in the pouch and may not be separately located on either the internal or external surface of the pouch.

In some embodiments, the at least one flavor glycoside may be positioned at a first position on an inner and/or external surface of the saliva permeable pouch by spraying or dropping the flavor glycoside onto the pouch. This may be achieved either before or after the oral product is incorporated within the pouch.

In some embodiments wherein the oral product further comprises one or more additional flavoring agents, the at least one flavor glycoside is positioned at a first position on an inner or external surface of the pouch and the one or more additional flavoring agents is positioned at a second position on an inner or external surface of the saliva permeable pouch, wherein the first and second positions are distinct from each other on the inner or external surface of the pouch. In some embodiments wherein the oral product further comprises one or more additional flavoring agents, the at least one flavor glycoside is positioned at a first position on an inner surface of the pouch and the one or more additional flavoring agents is positioned at a second position on an inner surface of the saliva permeable pouch, wherein the first and second positions are distinct from each other on the inner surface of the pouch. Such embodiments may result in a desirable product that delivers different flavors to the user when different parts of the product are in direct contact with the oral cavity of the user (e.g. in contact with the user’s tongue or taste buds).

In addition, separate location of the flavoring agents may allow differential release profiles (e.g., one flavoring agent may be rapidly available to the mouth and/or digestive system, and the other flavoring agent in the flavor glycoside may be released more gradually with product use). For example, in some embodiments, the composition within the pouched product may include at least one flavor glycoside, and at least one additional flavoring agent may be disposed in or on the inner or external surface of the pouched product (e.g. on or in the pouch material as disclosed herein).

The moisture content of the oral product may vary depending on the format in which the composition is provided. In some embodiments as described hereinabove, the oral product may be in the form of moist snuff or snus and/or may be provided in pouched formats. In some embodiments (e.g. for snus-type products), the moisture content of the composition (before insertion of the product into the user’s mouth) may be at least about 20% by weight, such as at least 30% by weight, such as at least 40% by weight, such as at least 50% by weight of the oral product. In some embodiments (for example, for snus-type products; e.g. non-pouched or pouched snus products), the moisture content of the composition (before insertion of the product into the user’s mouth) may be from about 20% to about 70% by weight, such as from about 30% to about 60% by weight, such as from about 40% to about 55% by weight of the oral product.

In some embodiments, the oral product may be a snus-type or snuff-type product that is in ‘dry’ form. In such embodiments, the moisture content of the oral product may be no greater than about 10% by weight, such as no greater than about 5% by weight of the oral product. For example, the moisture content may be from about 0.1% to about 10% by weight, such as from about 1% to about 5% by weight of the oral product.

Package

According to some embodiments described herein, there is provided a package containing an oral product as described herein. For example, the package may contain the oral product in powdered form. In such embodiments, the package may be in the form of a tin or plastic container. Alternatively or additionally, the package may contain the oral product in the form of a lozenge, pastille, tablet, or the like. The package may be in the form of a blister pack, tin or plastic container containing such oral dosage forms.

According to some embodiments described herein, there is provided a package containing at least one pouched oral product as described herein. A pouched product as described herein can be packaged within any suitable inner packaging material and/or outer container. See also, for example, the various types of containers for smokeless types of products that are set forth in U.S. Pat. Nos. 7,014,039 to Henson et al.; 7,537,110 to Kutsch et al.; 7,584,843 to Kutsch et al.; 8,397,945 to Gelardi et al., D592,956 to Thiellier; D594,154 to Patel et al.; and D625,178 to Bailey et al.; U.S. Pat. Pub. Nos. 2008/0173317 to Robinson et al.; 2009/0014343 to Clark et al.; 2009/0014450 to Bjorkholm; 2009/0250360 to Bellamah et al.; 2009/0266837 to Gelardi et al.; 2009/0223989 to Gelardi; 2009/0230003 to Thiellier; 2010/0084424 to Gelardi; and 2010/0133140 to Bailey et al; 2010/0264157 to Bailey et al.; and 2011/0168712 to Bailey et al. which are incorporated herein by reference. For example, the package may be a tin or plastic container which contains a plurality of the pouched oral products.

Process

In accordance with some embodiments described herein, there is provided an oral product as described herein, the process comprising the steps of:

(a) providing at least one filler or bulking agent and at least one flavor glycoside, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; and
(b) contacting the at least one filler or bulking agent and at least one flavor glycoside to provide the oral product.

The flavor glycoside may be as described hereinabove, and may be obtained using any of the methods described herein.

In some embodiments, the process further comprises the step of obtaining the at least one flavor glycoside by using a biotechnological process. The biotechnological process may be as described in detail hereinabove.

In some embodiments, the step (b) comprises mixing the at least one filler or bulking agent and the at least one flavor glycoside. In some embodiments, the at least one flavor glycoside is in solid form (e.g. in the form of a powder). The flavor glycoside may be mixed directly with the at least filler or bulking agent to provide the oral product. In some embodiments, the at least one flavor glycoside may be dissolved in a hydrophilic solvent (e.g. water and/or alcohol) prior to contacting the at least one filler or bulking agent. For example, the at least one flavor glycoside may be dissolved in water or alcohol (e.g. ethanol or propylene glycol) before being mixed with the filler or bulking agent. The process may, in such embodiments, comprise the step of drying the product so as to remove the solvent. For example, the product may be dried via heating, freeze-drying, spray-drying, or simply leaving the product at room temperature for a certain period of time. Preferably, the drying step comprises leaving the product at room temperature for a period of 1 hour to 48 hours to remove the solvent.

Use

According to some embodiments described herein, there is provided the use of a flavor glycoside to prolong flavor of an oral product, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond. In some embodiments, the flavor is delivered to the user for a period of at least about 10 minutes when the oral product is placed in the oral cavity of a user.

In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 15 minutes, such as at least about 15 minutes, such as at least about 20 minutes, such as at least about 25 minutes, such as at least about 30 minutes, such as at least about 40 minutes, such as at least about 50 minutes, such as at least about 60 minutes when the oral product is placed in the oral cavity of a user. In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 30 minutes when the oral product is placed in the oral cavity of a user. In some embodiments, the flavor from the flavoring agent is delivered to the user for a period of at least about 60 minutes when the oral product is placed in the oral cavity of a user. It has therefore been found that the use of a flavor glycoside in the oral product may prolong the delivery of flavor (from the flavoring agent) to the user during use.

According to some embodiments described herein, there is provided the use of a flavor glycoside to change the flavor released from an oral product over a period of time, wherein the oral product comprises a flavor glycoside and one or more additional flavoring agents, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond and wherein the one or more additional flavoring agents is distinct from the flavoring agent in the flavor glycoside.

In some embodiments, the one or more additional flavoring agents is released into the oral cavity of a user during use prior to the release of the flavoring agent from the flavor glycoside.

Further Broad Aspects

According to some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; wherein the flavoring agent is selected from the group consisting of geraniol, citronellol, nerol, maltol, ethylmaltol, fenchol, homofuraneol, furaneol, norfuraneol, 1-octen-3-ol, borneol, linalool, farnesol, hydroxycitronellol, 3,7-dimethyloctanol, myrcenol, lavandulol, nerolidol, terpineol, alpha-terpineol, menthol, thymol, carvacrol, myrtenol, carveol, santalol, piperitol, perillyl alcohol, patchouli alcohol, hexanol, 3-cis-hexanol, cis-3-hexen-1-ol, phenylethanol, eugenol, sesamol, sotolone, maple furanone, methyl anthranilate, guaiacol, raspberry ketone, 2-methoxy-4-vinylphenol, 4-ethylguaiacol, benzylalcohol, homofuraneol, and combinations thereof, and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes.

According to some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside; and (ii) at least one tobacco material; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; and wherein the at least one tobacco material is present in an amount of from about 0.001% to about 4.5% by weight of the oral product, and wherein the oral product is configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes.

According to some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; and wherein the at least one filler or bulking agent is or comprises microcrystalline cellulose. In some embodiments, the microcrystalline cellulose may be present in an amount of from about 45% to about 95% by weight of the oral product, such as from about 55% to about 95% by weight of the oral product, such as from about 55% to about 80% by weight of the oral product. In some embodiments, the oral product comprises microcrystalline cellulose in an amount of from about 45% to about 55% by weight of the oral product.

According to some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside, and (ii) at least one filler or bulking agent; wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond; and wherein the composition further comprises one or more additional flavoring agents that is not bound to a sugar via a glycosidic bond. In some embodiments, the one or more additional flavoring agents is different from the flavoring agent in the flavor glycoside. In some embodiments, the one or more additional flavoring agents is the same as the flavoring agent in the flavor glycoside. In some embodiments, the rate of release of the flavoring agent in the flavor glycoside is slower than the rate of release of the one or more additional flavoring agents, wherein the rate of release is measured as the rate at which a flavoring agent is released into a user’s mouth when the oral product is placed in the oral cavity of the user.

The flavor glycoside in such further broad aspects may be obtained by any means.

In accordance with some embodiments described herein, there is also provided an oral product comprising (i) at least one flavor glycoside; and (ii) at least one tobacco material, wherein the flavor glycoside comprises a flavoring agent bound to a sugar via a glycosidic bond, and wherein the flavor glycoside is obtained from a biotechnological process.

The above-described embodiments and disclosure equally applies to each of these further broad aspects. The combination of these further broad aspects with any of the embodiments described in the present disclosure is therefore specifically envisaged and encompassed by the present disclosure.

In all of the above embodiments, the oral product may be configured to be retained in the oral cavity of a user during use for a period of at least about 5 minutes.

EXAMPLES

Aspects of the present invention are more fully illustrated by the following examples, which are set forth to illustrate certain aspects of the present invention and are not to be construed as limiting thereof.

Example 1 - Oral Product Comprising Flavor Glycoside

In this Example, an oral product comprising microcrystalline cellulose was prepared in which a flavor glycoside was included, and the rates of release of the flavoring agent from the flavor glycoside were measured.

Materials and Methods

Chemicals - The following compounds were obtained commercially: 4-[4′-Hydroxy phenyl]-butan-2-one-4′-O-β-D-glucopyranoside (Raspberry ketone glycoside, >99%, 4GENE), ethanol (sensory grade, Sigma Aldrich), methanol (p.A., Sigma Aldrich), formic acid (>99%, Sigma Aldrich).

Test Product - An oral product was prepared based on the commercial product sold by British American Tobacco as LYFT. The product included microcrystalline cellulose as a filler. The product prepared differed from LYFT in that the product did not include any nicotine, and it was spiked with 1200 ppm of raspberry ketone glycoside dissolved in ethanol. The samples were then left at room temperature for one day to evaporate the ethanol.

Modern Oral Intervention Trial - Three healthy volunteers (female, 30-35 years of age) were asked to place the raspberry ketone spiked oral product under their upper lip, slightly to the side of the front teeth. After 30 sec, the product was moisturised by using the tip of the tongue. After 5 min, the product was disgorged into a centrifuge tube and any salivary enzymatic processes stopped by adding methanol (5 mL, p.A.) into the centrifuge tube. The procedure was repeated with intervention times of 0 (control sample), 10 and 20 min.

Sample Preparation - The centrifuge tubes containing the oral product and methanol (5 mL, p.A.) were vortexed for 10 sec and then placed on a plate shaker for 10 min to extract the remaining raspberry ketone glycoside. Afterwards, the tubes were centrifuged (13,200 rpm, 10 min) and the supernatant decanted into a safe lock tube (2 mL). The safe lock tube was centrifuged (13,200 rpm, 10 min) and an aliquot taken for analysis.

Quantitation of in-mouth glycoside degradation - Analysis was performed on a HPLC system (Knauer, Germany) equipped with a pump (AZURA P2.1S), auto-sampler (AZURA AS 6.1L) and diode-array UV/VIS detector (AZURA DAD 6.1L). Samples were loaded onto a reversed phase C18 column (Knauer, Eurospher II 100 C18, 3 mm × 100 mm) and separated with a 10 min gradient starting at 90% solvent mixture A (0.1% formic acid in water) in solvent mixture B (0.1% formic acid in methanol) at a flow rate of 0.5 mL/min changing to 30% A in 5 min and back to 90% A in 0.05 min holding for 4.5 min. The detection UV wavelength was set at 280 nm. Quantitation was performed by using an external calibration curve.

Results & Discussion

3 healthy volunteers were asked to place raspberry ketone spiked oral products under their upper lip according to a defined procedure and leave the pouch in their mouth for 5, 10 and 20 min. The remaining glycoside of the incubated samples as well as a control sample which was not incubated was extracted as described above and quantified by means of high pressure liquid chromatography and an external calibration curve. The resulting chromatogram of raspberry ketone glycoside at different incubation times is shown in FIG. 3.

The amount of flavoring agent in mg/ml remaining in the product at times 0, 5, 10 and 20 mins are shown in Table 1:

TABLE 1

amount of raspberry ketone remaining in product at various times after being placed in user’s mouths

Time
P-01
P-02
P-03

0 min
0.2
0.2
0.2

5 min
0.15
0.13
0.16

10 min
0.12
0.10
0.16

20 min
0.06
0.06
0.13

The amount of raspberry ketone thus released from the product at the various time periods was then calculated as a percentage based on the difference between the amount of raspberry ketone at time = 0 mins and the amount at t=5, 10 or 15 mins.

TABLE 2

percentage release of raspberry ketone from glycoside at various time intervals

Time
P-01
P-02
P-03

0 min
0%
0%
0%

5 min
25%
35%
20%

10 min
40%
50%
20%

20 min
70%
70%
35%

The results are also shown in FIG. 4. As illustrated in FIG. 4, a significant decrease in flavor glycoside could be observed upon in mouth incubation across all panellists with 35 - 70% of the glycoside being enzymatically cleaved after 20 min.

The various embodiments described herein are presented only to assist in understanding and teaching the claimed features. These embodiments are provided as a representative sample of embodiments only, and are not exhaustive and/or exclusive. It is to be understood that advantages, embodiments, examples, functions, features, structures, and/or other aspects described herein are not to be considered limitations on the scope of the invention as defined by the claims or limitations on equivalents to the claims, and that other embodiments may be utilized and modifications may be made without departing from the scope of the claimed invention. Various embodiments of the invention may suitably comprise, consist of, or consist essentially of, appropriate combinations of the disclosed elements, components, features, parts, steps, means, etc., other than those specifically described herein. In addition, this disclosure may include other inventions not presently claimed, but which may be claimed in future.

ORAL PRODUCT

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