Oral compositions, oral use articles, drinks and related applications

Abstract

The application discloses an oral composition, a mouth product, a drink and related applications of the oral composition. The oral composition provided by the application comprises a nicotine dehydrogenase, probiotics and food-grade synergistic fiber, the nicotine dehydrogenase is derived from Pseudomonas putida, and the probiotics are Lactobacillus plantarum. The oral composition provided by the application has a nicotine degradation rate of 90.5% in a simulation system. Meanwhile, the composition has excellent processability, gastrointestinal stability, high food compatibility and bioavailability.

Description

Technical Field

[0001] This application relates to the field of smokeless product technology, specifically to an oral composition, a lozenge, a beverage, and related applications of the oral composition. Background Technology

[0002] With increasing health awareness, functional food or health supplement ingredients that address nicotine dependence and reduce the harmful effects of nicotine intake have become a research hotspot. Currently, the main technical pathways for nicotine degradation include: chemical catalysis, microbial fermentation, and free enzyme preparations. Chemical catalysis uses metal catalysts, but suffers from biocompatibility issues and is unsuitable for foods and health supplements. Microbial fermentation can utilize certain strains to degrade nicotine, but it is inefficient and time-consuming (degradation cycle > 48 hours). Free enzyme preparations include enzymes such as nicotine dehydrogenase, which can be used directly. Nicotine dehydrogenase is an enzyme that catalyzes the redox reaction of nicotine and belongs to the dehydrogenase class. It participates in the metabolism of nicotine by converting nicotine into metabolites such as cotinine. This enzyme has high activity in the liver and is one of the key enzymes in nicotine metabolism, but it is easily inactivated in the acidic environment of the stomach. Therefore, developing more effective functional foods or health supplements remains an urgent problem to be solved. Summary of the Invention

[0003] Based on this, this application provides an oral composition and its application, as well as oral articles and beverages containing the oral composition.

[0004] This application provides an oral composition comprising nicotinic dehydrogenase, probiotics, and food-grade synergistic fiber, wherein the nicotinic dehydrogenase is derived from *Pseudomonas putida* and the probiotics are *Lactobacillus plantarum*.

[0005] In some specific embodiments, the aforementioned food-grade enhancing fibers include at least one of β-glucan, pectin, inulin, and chitosan.

[0006] In some specific embodiments, the oral composition includes nicotine dehydrogenase lyophilized agent and / or probiotic lyophilized agent.

[0007] In some specific embodiments, the above-mentioned nicotine dehydrogenase lyophilizer and probiotic lyophilizer also include a lyophilization protective substance, which includes at least one of trehalose, skim milk powder, sucrose and monosodium glutamate.

[0008] In some specific embodiments, the oral composition comprises at least one of nicotine dehydrogenase inclusion complex, probiotic inclusion complex, and nicotine dehydrogenase-probiotic inclusion complex.

[0009] In some specific embodiments, the nicotine dehydrogenase inclusion complex, probiotic inclusion complex, and nicotine dehydrogenase-probiotic inclusion complex include cyclodextrin inclusion substances, which include at least one of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and methyl-β-cyclodextrin.

[0010] In some specific embodiments, the oral composition further includes an enteric coating shell for containing nicotinic dehydrogenase and / or probiotics.

[0011] In some specific embodiments, the specific activity of the nicotinic dehydrogenase in the oral composition is not less than 500 U / mg.

[0012] In some specific embodiments, the number of live bacteria in the probiotic raw material of the oral composition is not less than 1×10^11 CFU / g.

[0013] In some specific embodiments, in a simulated gastric fluid environment at pH=2, the enzyme activity retention rate of nicotine dehydrogenase in the above oral composition is greater than or equal to 80% within two hours.

[0014] In some specific embodiments, in a simulated gastric fluid environment with pH=2, the viable survival rate of the probiotics in the above oral composition is greater than or equal to 85% within two hours.

[0015] This application also provides the application of any of the above-mentioned oral compositions in the field of nicotine-reducing foods and / or health products.

[0016] This application also provides a lozenge comprising any of the above-described oral compositions.

[0017] In some specific embodiments, the above-mentioned oral products include at least one of oral dissolving films, chewing gum, oral tablets, or oral pouches.

[0018] This application also provides a beverage comprising any of the above-described oral compositions.

[0019] The beneficial effects of this application are as follows: This application provides an oral composition containing nicotine dehydrogenase, probiotics, and food-grade synergistic fiber. In a simulated system, this oral composition achieves a nicotine degradation rate of up to 90.5%. Through the synergistic effect of the food-grade synergistic fiber, the oral composition maintains an effective degradation concentration for more than 24 hours in an intestinal model. This oral composition has high food compatibility and bioavailability, uses all food-grade ingredients, and in animal models, its relative bioavailability in reducing nicotine absorption is 2.8 times higher than that of traditional simple mixed formulations. Detailed Implementation

[0020] The present application will now be described in further detail through specific embodiments. In the following embodiments, many details are described to facilitate a better understanding of the present application. However, those skilled in the art will readily recognize that some features may be omitted in different situations, or may be replaced by other elements, materials, or methods. In some cases, certain operations related to the present application are not shown or described in the specification. This is to avoid obscuring the core parts of the present application with excessive description. For those skilled in the art, detailed description of these related operations is not necessary; they can fully understand the related operations based on the description in the specification and general technical knowledge in the art.

[0021] Furthermore, the features, operations, or characteristics described in the specification can be combined in any suitable manner to form various embodiments. At the same time, the steps or actions in the method description can be rearranged or adjusted in a manner obvious to those skilled in the art. Therefore, the various orders in the specification are only for clearly describing a particular embodiment and do not imply a necessary order, unless otherwise stated that a particular order must be followed.

[0022] The serial numbers assigned to components in this document, such as "first" and "second," are used only to distinguish the described objects and have no sequential or technical meaning. The terms "connection" and "linkage" used in this application, unless otherwise specified, include both direct and indirect connections (linkages).

[0023] As mentioned earlier, existing technologies for directly degrading nicotine components (active ingredients) face significant challenges when applied to oral foods and health supplements. Furthermore, food processing techniques (such as high temperatures and homogenization) can affect the stability of active ingredients, making it difficult for existing formulation processes to balance high efficiency with the feasibility of food production. For example, oral enzyme preparations (such as nicotine dehydrogenase) are easily inactivated in the acidic environment of the stomach, and their activity retention rate in simulated gastric juice is typically less than 50%, limiting their application in food. Meanwhile, single probiotic strains have limited efficiency in degrading nicotine, with in vitro degradation rates typically below 40%, making it difficult to achieve efficient degradation in complex food systems and the intestinal environment. Therefore, existing technologies lack a systematic solution suitable for the food or health supplement industry that can maintain the high stability and sustained activity of active ingredients in the gastrointestinal tract.

[0024] To address the aforementioned issues, this application provides an oral composition of a ternary synergistic system of "nicotine dehydrogenase-probiotics-food-grade synergistic fiber" that can be efficiently applied in the food or health product field, achieving efficient and stable nicotine degradation in food form. In the oral composition, the nicotine dehydrogenase is derived from *Pseudomonas putida*, and the probiotic is *Lactobacillus plantarum*. The nicotine dehydrogenase uses raw materials with a specific activity ≥ 500 U / mg. The probiotics use raw materials with a viable count ≥ 1 × 10^11 CFU / g, and the viable count in the oral composition generally reaches 2 × 10^7 CFU / mL; in some embodiments, the viable count in the oral composition can reach as high as 5 × 10^10 CFU / g.

[0025] In this application, food-grade synergistic fibers are a type of fiber that can enhance the functionality of food. Food-grade synergistic fibers suitable for this application include at least one of β-glucan, pectin, inulin, and chitosan. In the oral composition of this application, food-grade synergistic fibers can effectively promote the proliferation and colonization of probiotics and prolong the overall duration of action by increasing the viscosity of intestinal contents and the prebiotic effect. Specifically, after dissolving in water, food-grade synergistic fibers form a viscous gel network that "anchors" nicotine dehydrogenase and probiotics within it, prolonging their residence and action time in the intestine; simultaneously, food-grade synergistic fibers can be fermented and utilized by *Lactobacillus plantarum* as a prebiotic, promoting its proliferation; furthermore, some food-grade synergistic fibers suitable for this application (such as chitosan) can also physically adsorb nicotine, assisting in its degradation. In some specific embodiments, the food-grade synergistic fiber is β-glucan (derived from oats), with a purity ≥ 90% and a viscosity (1% aqueous solution, 20°C) in the range of 200-500 mPa•s.

[0026] In the oral composition provided in this application, each component may be in the same or different specific dosage forms.

[0027] In some embodiments, nicotinic dehydrogenase and / or probiotics may be in lyophilized dosage form, i.e., the oral composition includes a lyophilized nicotinic dehydrogenase and / or a lyophilized probiotic. The lyophilized nicotinic dehydrogenase includes nicotinic dehydrogenase and a lyophilization protectant, and the lyophilized probiotic includes probiotics and a lyophilization protectant. The lyophilization protectant in the lyophilized agent is used to maintain the activity of nicotinic dehydrogenase and probiotics during the freeze-drying process. In some embodiments, the lyophilization protectant in the lyophilized agent is composed of trehalose, skim milk powder, sucrose, and monosodium glutamate, in the following mass parts: trehalose 15-25 parts, skim milk powder 8-12 parts, sucrose 5-8 parts, and monosodium glutamate 2-4 parts.

[0028] In some specific embodiments, nicotinic dehydrogenase and / or probiotics may be in the form of inclusion complex dosage forms, i.e., the oral composition includes at least one of nicotinic dehydrogenase inclusion complex, probiotic inclusion complex, and nicotinic dehydrogenase-probiotic inclusion complex. In actual preparation, by encapsulating nicotinic dehydrogenase, probiotics, nicotinic dehydrogenase lyophilized powder, and / or probiotic lyophilized powder in a cyclodextrin inclusion complex, it is beneficial to maintain the activity of nicotinic dehydrogenase. Its mechanism of action includes: forming a molecular capsule to isolate the enzyme from the erosion of gastric acid and pepsin; improving the thermal stability of the enzyme and enhancing its resistance to denaturation during processes such as pasteurization; and masking any possible off-odors. In some specific embodiments, the above-mentioned cyclodextrin inclusion complex contains at least one of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and methyl-β-cyclodextrin. In some specific embodiments, the cyclodextrin inclusion substances in the nicotine dehydrogenase inclusion complex, probiotic inclusion complex, and nicotine dehydrogenase-probiotic inclusion complex are composed of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and methyl-β-cyclodextrin. Based on the mass fractions of each cyclodextrin inclusion substance in the oral composition, β-cyclodextrin is 8 to 15 parts, hydroxypropyl-β-cyclodextrin is 3 to 8 parts, and methyl-β-cyclodextrin is 1 to 3 parts.

[0029] In some embodiments, the oral composition of this application further includes an enteric coating shell for encapsulating nicotinic dehydrogenase and / or probiotics. Specific dosage forms of the nicotinic dehydrogenase and / or probiotics include, but are not limited to, lyophilized agents and inclusion complexes. In some embodiments, the enteric coating shell can be a macroscopic shell, such as the enteric coating of a lozenge, the shell of a capsule, and may also encapsulate food-grade synergistic fibers, as well as other components that the oral composition may include, such as fillers, flavoring agents, sustained-release agents, pH adjusters, etc. In some embodiments, the enteric coating shell can also be a microscopic shell, such as the shell of an enteric microcapsule, which is mainly used to encapsulate nicotinic dehydrogenase and / or probiotics, to correspondingly form nicotinic dehydrogenase enteric microcapsules, probiotic enteric microcapsules, or nicotinic dehydrogenase-probiotic enteric microcapsules. Different packaging methods do not constitute a limitation on the enteric coating shell of the oral composition of this application. In some specific embodiments, the enteric coating of the oral composition comprises hydroxypropyl methylcellulose phthalate, Eutrapeptide L100-55, and triethyl citrate (plasticizer). Based on the mass parts of the final product, it includes 6 to 10 parts of hydroxypropyl methylcellulose phthalate, 4 to 8 parts of Eutrapeptide L100-55, and 1 to 2 parts of triethyl citrate.

[0030] The above-mentioned oral composition can be applied in the field of nicotine-lowering foods and / or health products, for example, by processing it into food or health products.

[0031] In some specific implementations, the types of food or health products that can be processed include, but are not limited to, solid and liquid forms.

[0032] In some specific embodiments, the solid-type food or health product containing the composition described above may be a lozenge. The preparation parameters for this lozenge are as follows:

[0033] Tableting pressure: 8~12 kN;

[0034] Tablet weight: 500±50 mg;

[0035] Hardness: 60~80 N.

[0036] The quality standards for this lozenge can be referenced as follows:

[0037] Enzyme activity: 90.0% to 110.0% of the labeled amount (the theoretical content of the substance per unit of formulation (e.g., per tablet) as stated on the label / instructions for use);

[0038] Viable bacteria count: ≥ 1.0 × 10^9 CFU / tablet;

[0039] β-glucan content: 85.0~115.0% of the labeled amount;

[0040] Content uniformity: RSD ≤ 5.0%;

[0041] Enteric coating properties: ≤ 10% release rate in artificial gastric juice (pH 1.2) after 2 hours.

[0042] The preparation method of the above-mentioned lozenges includes: first preparing nicotine dehydrogenase-cyclodextrin inclusion complex, then mixing it with probiotic freeze-dried powder, β-glucan, filler, etc. for granulation, compressing into tablets, and then coating with enteric coating.

[0043] In some specific embodiments, the liquid-type food or health product containing the composition described above can be a functional beverage, and the preparation parameters of the beverage are as follows:

[0044] pH range: 4.2~4.8 (balancing stability and taste);

[0045] Soluble solids: 12~15 °Brix;

[0046] Viscosity: 180~350 mPa•s (20℃, adjusted by β-glucan).

[0047] The quality standards for this functional beverage can be referenced as follows:

[0048] pH value: 4.2~4.8;

[0049] Enzyme activity: 85.0%–115.0% of the labeled amount;

[0050] viable bacteria count: ≥ 1.0 × 10^8 CFU / mL;

[0051] β-glucan content: 80.0%~120.0% of the labeled amount;

[0052] Viscosity: 180~350 mPa•s (20℃);

[0053] Microbial limits: Compliant with GB 7101 "National Food Safety Standard for Beverages".

[0054] The preparation method of the above-mentioned functional beverages includes: dissolving β-glucan, adding nicotinic acid dehydrogenase, probiotics and other ingredients, homogenizing, pasteurizing and filling.

[0055] In some specific embodiments of this application, a lozenge comprising the above-described oral composition is provided. The lozenge uses the oral composition as the main active ingredient, and its specific product form includes at least one of oral dissolving film, chewing gum, lozenge tablets, or lozenge pouches.

[0056] In some specific embodiments, the oral composition includes a film-forming agent, which itself has a film-like structure or can be directly prepared into an orally disintegrating film. In some specific embodiments, the oral composition does not include a film-forming agent, and an orally disintegrating film can be prepared by adding it to a film-forming solution. It should be noted that the final product form of orally disintegrating film-type oral products can be diverse. In some embodiments, the orally disintegrating film consists of one or more film-like structures including the oral composition; in some embodiments, in addition to having one or more film-like structures including the oral composition, the orally disintegrating film may also have a support layer disposed on at least one surface of the film-like structure. For example, in some embodiments, the specific structure of the orally disintegrating film-type oral product is nonwoven fabric-functional layer-isolation layer-functional layer-nonwoven fabric, wherein the nonwoven fabric is a liquid-permeable support layer, the functional layer is a film-like structure including the oral composition, and the isolation layer is usually a water-repellent fabric layer used to physically isolate two adjacent functional layers.

[0057] In some specific embodiments, the mouth bag is a product that encapsulates a mouth-containing composition within a liquid-permeable bag. The liquid-permeable bag is permeable to saliva; for example, the liquid-permeable bag can be a non-woven bag. The encapsulated contents can be in the form of powder, granules, flakes, film, or other structures.

[0058] In some embodiments, the oral composition includes a gum base, which can be directly prepared into chewing gum. In other embodiments, the oral composition does not include a gum base, but is used in combination with a gum base and other raw materials to prepare chewing gum.

[0059] In some specific embodiments, the lozenge is a compressed tablet containing an oral composition that dissolves slowly in the oral cavity.

[0060] Example 1: Oral Composition 1, Preparation and Comparative Experiment

[0061] 1. Preparation of oral composition 1

[0062] 1) Preparation of nicotine dehydrogenase-cyclodextrin inclusion complex: Nicotine dehydrogenase (derived from Pseudomonas putida, specific activity ≥500 U / mg) was dissolved in phosphate buffer at pH 6.0. β-cyclodextrin, hydroxypropyl-β-cyclodextrin and methyl-β-cyclodextrin were added at a mass ratio of 1:3:1:0.5 (enzyme: β-cyclodextrin: hydroxypropyl-β-cyclodextrin: methyl-β-cyclodextrin). The mixture was stirred at 4°C for 4 hours for inclusion and then freeze-dried to obtain lyophilized powder of nicotine dehydrogenase-cyclodextrin inclusion complex.

[0063] 2) Preparation of probiotic powder: The fermentation broth of Lactobacillus plantarum is centrifuged to collect the bacterial sludge, and freeze-drying agents (trehalose, skim milk powder, sucrose, monosodium glutamate) are added. After mixing, the mixture is freeze-dried under vacuum to obtain highly active probiotic freeze-dried powder.

[0064] 3) Total mixing: The above-mentioned nicotine dehydrogenase-cyclodextrin inclusion complex lyophilized powder, probiotic lyophilized powder and food-grade synergistic fiber (β-glucan) are mixed evenly in a three-dimensional mixer, and then packaged with nitrogen to obtain oral composition 1.

[0065] 2. Experimental results of oral composition 1

[0066] 1) The results of the stability test are shown in Table 1.

[0067] Table 1. Stability data of each component of oral composition 1

[0068] Storage conditions time Enzyme activity retention rate viable bacteria survival rate Degradation rate maintained 4℃ 3 months 95.2 ± 1.2% 92.3 ± 2.1% 94.1 ± 1.5% 25℃ / 60%RH 3 months 87.6 ± 2.3% 84.1 ± 2.8% 88.5 ± 2.1% 37℃ / 75%RH 1 month 78.9 ± 3.1% 75.2 ± 3.5% 81.9 ± 2.8%

[0069] The experimental results show that oral composition 1 exhibits excellent long-term storage stability under refrigeration at 4°C, with enzyme activity retention and viable bacterial survival rates both exceeding 90%. Even after 3 months of storage at 25°C, the degradation of key indicators remained within acceptable limits, indicating that this oral composition possesses certain potential for room temperature storage and transportation.

[0070] 2) In vitro degradation kinetic parameters

[0071] Maximum degradation rate (V) max ): 8.15±0.40 μmol / min

[0072] Michaelis constant (K) m ): 12.5 ± 1.2 μM

[0073] Catalytic efficiency (k cat / K m ): (6.65±0.38)×10^4 m -1·s -1

[0074] Experimental results indicate that the kinetic parameters show that the nicotine dehydrogenase in this composition has a high affinity for the substrate nicotine (K). m (lower value) and high catalytic efficiency (k cat / K m (High value). V max The value reflects the strong potential degradation capacity of this enzyme system, providing a foundation for subsequent efficient in vivo degradation.

[0075] Example 2: Verification of the synergistic degradation effect between nicotine dehydrogenase and probiotics

[0076] This example compares nicotine dehydrogenase and probiotics. The experimental conditions and procedures are as follows:

[0077] Grouping: Three experimental groups were set up, with three parallel groups in each group.

[0078] Group A (enzyme group): Contains only nicotinic dehydrogenase (derived from Pseudomonas putida, final concentration 5 U / mL).

[0079] Group B (bacterial group): Contains only Lactobacillus plantarum (final concentration 1×10^8 CFU / mL).

[0080] Group C (enzyme + bacteria group): Contains both nicotinic acid dehydrogenase (5 U / mL) and Lactobacillus plantarum (1×10^8 CFU / mL).

[0081] Reaction system: Nicotine standard was added to a final concentration of 1 μg / mL in a simulated intestinal environment (MRS medium, pH 6.5, 37℃).

[0082] Detection: Samples were taken at 0h and 24h, filtered through a microporous membrane, and the nicotine residue was determined by high performance liquid chromatography, and the degradation rate was calculated.

[0083] Synergy Index Calculation: Synergy Enhancement Index = Measured Combined Degradation Rate / (Enzyme Degradation Rate Alone + Bacterial Degradation Rate Alone - Enzyme Degradation Rate Alone × Bacterial Degradation Rate Alone). An index > 1 indicates synergy.

[0084] Experimental results showed that the degradation rate of nicotine dehydrogenase alone was 68.5%, and that of probiotics alone was 38.2%. The degradation rate of the mixture of nicotine dehydrogenase and probiotics was 82.1%. Therefore, the synergistic effect index of nicotine dehydrogenase and probiotics was 1.31. This in vitro synergistic experiment confirmed that nicotine dehydrogenase exhibited a significant synergistic effect with probiotics at a final concentration of 5 U / mL, and this concentration can be used as a reference for the effective dosage of enzyme in the composition.

[0085] Example 3: Synergistic effect of food-grade synergistic fiber (β-glucan)

[0086] 1. Comparative experiments using simulated gut fermentation models

[0087] Oral composition 2 was prepared using the same method as oral composition 1 in Example 1, except that oral composition 2 does not contain β-glucan.

[0088] Oral composition 1 and oral composition 2 were compared in a simulated colonic fermentation model. To verify the synergistic effect of β-glucan, equal amounts of oral composition 1 containing β-glucan (experimental group) and oral composition 2 without β-glucan (control group) were inoculated into the simulated colonic fermentation model, respectively. Samples were taken at 0h, 12h, 24h, and 36h of fermentation to determine the viable cell count and nicotine degradation capacity.

[0089] Experimental results showed that the number of live probiotics in oral composition 1 was 1.5 orders of magnitude higher than that in oral composition 2 at 24 hours. The duration of nicotine degradation ability of oral composition 1 was extended by about 80% compared with that of oral composition 2, proving that β-glucan significantly enhanced the colonization ability of probiotics and the long-term effectiveness of the system.

[0090] 2. Animal experiments

[0091] 1) A rat model (n=40) was established. A quantitative amount of nicotine solution was administered by gavage, and the rats were divided into three groups: control group, experimental group 1, and experimental group 2.

[0092] Control group: Administered an equal volume of physiological saline.

[0093] Experimental group 1 (enzyme + bacteria): nicotine dehydrogenase and probiotic mixed lyophilized powder without synergistic fiber were administered.

[0094] Experimental Group 2 (Full Composition): The lyophilized powder of the "nicotine dehydrogenase-probiotics-food grade gain fiber" composition containing β-glucan of this application was given.

[0095] 2) The results are shown in Table 2 (mean ± SD):

[0096] Table 2 Animal Experiment Results

[0097] Group Blood drug concentration AUC (ng·h / mL) Intestinal nicotine residue rate (24h) Quantitative analysis of probiotics in feces (log CFU / g) control group 125.6 ± 8.9 100% Not detected Experimental Group 1 (Enzyme + Bacteria) 65.2 ± 5.2 45.2% 7.8 ± 0.3 Experimental Group 2 (Full Composition) 42.5 ± 3.8 28.7% 9.2 ± 0.4

[0098] Conclusion: Compared with experimental group 1, which did not contain synergistic fiber, experimental group 2, which contained "nicotine dehydrogenase-probiotics-food-grade synergistic fiber", significantly reduced blood nicotine exposure (AUC decreased by 34.8%), more effectively reduced intestinal nicotine residue, and significantly promoted the survival and colonization of probiotics in the intestine.

[0099] Example 4: Verification of the protective effect of cyclodextrin inclusion complex

[0100] In this example, three comparative experiments were conducted. The experimental steps and conditions are as follows:

[0101] Comparative Example 1 used only nicotinic dehydrogenase (derived from Pseudomonas putida).

[0102] In Comparative Example 2, β-cyclodextrin was used to encapsulate nicotinic dehydrogenase, with a mass ratio of β-cyclodextrin to nicotinic dehydrogenase of 3:1. The encapsulation method was as follows: both were dissolved in water, stirred at 4°C for 4 hours, and then freeze-dried.

[0103] In Comparative Example 3, a complex cyclodextrin inclusion system was used to include nicotine dehydrogenase, wherein the mass ratio of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, methyl-β-cyclodextrin to nicotine dehydrogenase was 3:1:0.5:0.3. The inclusion method was the same as in Comparative Example 2.

[0104] In this example, a comparative experiment was conducted on the cyclodextrin inclusion system in simulated gastric fluid (pH 2.0, 2h). To verify the protective effect of different inclusion systems, three enzyme preparations were prepared: Comparative Example 1 was a solution of free nicotine dehydrogenase without any inclusion; Comparative Example 2 was an enzyme-β-CD inclusion complex encapsulated only with β-cyclodextrin; and the experimental group was an enzyme-complex cyclodextrin inclusion complex encapsulated using a complex cyclodextrin system. The three samples were incubated in simulated gastric fluid (pH 2.0, containing pepsin) at 37°C for 2 hours. Enzyme activity (U / mL) was measured before and after incubation. Activity retention rate (%) = (enzyme activity after incubation / enzyme activity before incubation) × 100%.

[0105] The experimental results showed that under the experimental conditions, the activity retention rate of nicotinic dehydrogenase was 45.2% without inclusion complex. The activity retention rate was 72.8% with only β-cyclodextrin inclusion complex. Furthermore, the activity retention rate was 84.3% with complex cyclodextrin inclusion complex.

[0106] Example 5: Preparation of Nicotine-Lowering Functional Tablets

[0107] 1. The formulation (core) is as follows:

[0108] Nicotine dehydrogenase lyophilized powder (derived from *Pseudomonas putida*): 125 mg (250 U);

[0109] Lactobacillus plantarum lyophilized powder: 150 mg (7.5×10^9 CFU);

[0110] β-glucan (food grade): 20 mg;

[0111] β-Cyclodextrin: 45 mg;

[0112] Trehalose: 75 mg;

[0113] Microcrystalline cellulose: 85 mg;

[0114] Cross-linked carboxymethyl cellulose sodium: 12 mg;

[0115] Magnesium stearate: 5 mg.

[0116] 2. Preparation process

[0117] 1) Preparation of enzyme-cyclodextrin inclusion complex: β-cyclodextrin and nicotine dehydrogenase were included in an aqueous solution at a mass ratio of 3:1 and then spray-dried.

[0118] 2) Mixing and granulation: Thoroughly mix the inclusion complex, probiotic freeze-dried powder, β-glucan, trehalose, microcrystalline cellulose, etc. from step 1), wet granulate with an appropriate amount of pure water as a binder (18% of the amount), and dry at 40°C.

[0119] 3) Tableting: Add magnesium stearate and other excipients, mix well, and then compress into tablets. Control the tablet weight to 500 mg, the pressure to 10 kN, and the hardness to 70 N.

[0120] 4) Preparation and Coating of Enteric Coating Solution: Hydroxypropyl methylcellulose phthalate (HPMCP) and Eutectic L100-55 were dissolved in an ethanol / water mixture, and triethyl citrate was added as a plasticizer to prepare a coating solution with a solid content of 10%. The tablet cores were placed in a high-efficiency coating pan, with an inlet air temperature of 40°C, a tablet bed temperature of 38-40°C, and a spray gun pressure of 1.5 bar for coating. The coating weight gain was 8%.

[0121] The disintegration time (enteric-coated) of the lozenge in this example is: no disintegration in artificial gastric fluid for 2 hours, and complete disintegration within 45 minutes in artificial intestinal fluid.

[0122] Example 6: Preparation of a Nicotine-Lowering Functional Beverage

[0123] 1. Formula composition (per liter):

[0124] Nicotine dehydrogenase (derived from *Pseudomonas putida*): 15,000 U;

[0125] Lactobacillus plantarum: 2×10^10 CFU;

[0126] β-glucan: 5 g;

[0127] Hydroxypropyl-β-cyclodextrin: 8 g;

[0128] Trehalose: 25 g;

[0129] High-fructose corn syrup: 80 g;

[0130] Citric acid: 2 g;

[0131] Natural spices: as needed.

[0132] 2. Process parameters:

[0133] 1) Premixing: Dry mix β-glucan with some trehalose, slowly add it to pure water, and stir until completely dissolved.

[0134] 2) Mixing: Add the remaining ingredients, including enzymes, probiotics (added later), hydroxypropyl-β-cyclodextrin, etc., and gently stir to mix.

[0135] 3) Homogenization: Homogenize under 25 MPa pressure to ensure the system is uniform and stable.

[0136] 4) Pasteurization: Sterilize at 72℃ for 15 seconds.

[0137] 5) Filling and storage: Aseptic filling at 25℃, and refrigerated storage at 4℃.

[0138] The live bacteria survival rate in the functional beverage provided in this example is ≥ 85% after storage at 4℃ for 30 days.

[0139] The above examples illustrate this application only to aid understanding and are not intended to limit its scope. Those skilled in the art to which this application pertains can make various simple deductions, modifications, or substitutions based on the ideas presented.

Claims

1. An oral composition, characterized in that, The oral composition comprises nicotinic dehydrogenase, probiotics, and food-grade synergistic fiber, wherein the nicotinic dehydrogenase is derived from *Pseudomonas putida*, and the probiotics are *Lactobacillus plantarum*.

2. The oral composition according to claim 1, characterized in that, The food-grade enhancement fiber includes at least one of β-glucan, pectin, inulin, and chitosan.

3. The oral composition according to claim 1, characterized in that, The oral composition comprises a lyophilized nicotinic dehydrogenase and / or a lyophilized probiotic; and / or, The oral composition comprises at least one of nicotine dehydrogenase inclusion complex, probiotic inclusion complex, and nicotine dehydrogenase-probiotic inclusion complex.

4. The oral composition according to claim 3, characterized in that, The nicotine dehydrogenase lyophilizer and the probiotic lyophilizer include a lyophilization protectant, which includes at least one selected from trehalose, skim milk powder, sucrose, and monosodium glutamate; and / or, The nicotine dehydrogenase inclusion complex, the probiotic inclusion complex, and the nicotine dehydrogenase-probiotic inclusion complex include cyclodextrin inclusion substances, wherein the cyclodextrin inclusion substances include at least one of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, and methyl-β-cyclodextrin.

5. The oral composition according to claims 1-4, characterized in that, The oral composition further includes an enteric coating shell for containing the nicotinic dehydrogenase and / or the probiotics.

6. The oral composition according to any one of claims 1 to 5, characterized in that, The specific activity of the nicotinic dehydrogenase is greater than or equal to 500 U / mg; and / or, The probiotic raw material has a live bacteria count greater than or equal to 1 × 10^11 CFU / g; and / or, In a simulated gastric fluid environment at pH=2, the enzyme activity retention rate of the nicotinic dehydrogenase in the oral composition is greater than or equal to 80% over two hours; and / or, In a simulated gastric fluid environment at pH=2, the viable bacterial survival rate of the probiotics in the oral composition is greater than or equal to 85% within two hours.

7. The use of the oral composition according to any one of claims 1 to 6 in the field of nicotine-reducing foods and / or health products.

8. A mouth-held product, characterized in that, The oral product comprises the oral composition as described in any one of claims 1 to 6.

9. The oral article according to claim 8, characterized in that, The oral products include at least one of oral films, chewing gum, lozenges, or lozenge pouches.

10. A beverage, characterized in that, The beverage comprises the oral composition as described in any one of claims 1 to 6.