A bionic fermentation type sleep-aiding nutraceutical composition and a preparation method thereof

Abstract

The application discloses a kind of bionic fermentation type sleep-aiding medicinal and edible homologous composition and preparation method thereof, composition includes jujube kernel, longan meat, lily, poria cocos, black sesame, lotus seed, jujube, liquorice, oyster, dogwood, peach kernel, lophatherum gracile;When preparing, jujube kernel, peach kernel, black sesame are separated after bionic fermentation by composite probiotics;Residue and lily, lotus seed, poria cocos, lophatherum gracile are alcohol-extracted;Drug residue and longan meat, jujube, liquorice, dogwood are water-extracted;Fermentation supernatant and alcohol extract are prepared into immediate-release component, and water extract is prepared into sustained-release component, then mixed into ultrafine powder with oyster and formed;The application realizes saponin biotransformation by fermentation, avoids component antagonism by step extraction, matches sleep phase by immediate-release / sustained-release dual-phase release, has intestinal-brain axis regulation and multi-target nerve-calming effect, and has high bioavailability, quick effect, long maintenance time and good safety.

Description

Technical Field

[0001] This invention belongs to the field of food or health food technology, specifically relating to a sleep-aid composition prepared from medicinal and edible substances through biomimetic fermentation and phase separation extraction processes, as well as a method for preparing the composition. Background Technology

[0002] Insomnia and declining sleep quality have become a global public health problem. Epidemiological surveys indicate that the incidence of insomnia among adults in my country is approximately 38.2%, with chronic insomnia affecting over 10% of these individuals. Long-term insomnia not only leads to daytime dysfunction and cognitive decline but is also closely related to anxiety, depression, cardiovascular disease, and metabolic syndrome. Therefore, developing safe, effective, and long-term sleep aids is of significant social importance.

[0003] Existing traditional Chinese medicine compound formulas or health food compositions for improving sleep often rely on non-food and medicinal herbs as raw materials, resulting in insufficient safety and daily applicability. The classic formula Suanzaoren Tang (Ziziphus jujuba seed decoction) comes from the *Jinkui Yaolue* (Essential Prescriptions of the Golden Chamber), composed of ziziphus jujuba seed, anemarrhena asphodeloides, poria cocos, chuanxiong rhizome, and licorice root. Anemarrhena asphodeloides is cold in nature and moistening, and prolonged use can easily damage the spleen and stomach; chuanxiong rhizome is pungent and warm, and should be avoided by those with yin deficiency and excessive fire. Neither of these herbs is listed in the "both food and medicinal material" catalog, resulting in a narrow safety window for long-term use, making them unsuitable for promotion as daily health foods. While some commercially available calming health foods use ziziphus jujuba seed as the main ingredient, they often combine it with herbs such as polygala tenuifolia and albizia bark. The saponins in polygala tenuifolia can irritate the gastrointestinal mucosa, and sensitive individuals are prone to adverse reactions such as nausea and diarrhea. On the other hand, current sleep aids formulated solely with substances that are both food and medicine, such as single jujube seed extract preparations or Poria cocos extract capsules, generally suffer from simple formulations and limited efficacy, making it difficult to address the complex pathogenesis of insomnia, which involves multiple factors such as "deficiency of both heart and spleen, yin deficiency and yang hyperactivity, and blood stasis in the brain."

[0004] At the extraction process level, existing traditional Chinese medicine sedative compositions mostly employ a "one-pot" water or alcohol extraction method, which presents a structural contradiction between the transformation and retention of active ingredients. The core sedative components, such as jujube seed saponins A and B, exist primarily in glycoside form, with large molecular weights and poor lipid solubility, making it difficult to directly cross the blood-brain barrier to exert GABAergic regulatory effects. Studies have shown that these saponins require hydrolysis by β-glucuronidase secreted by intestinal flora to be converted into aglycones before they can be efficiently absorbed. Traditional water or alcohol extraction processes only achieve physical dissolution, failing to simulate the intestinal microecological environment for biotransformation, resulting in an actual bioavailability of less than 15% after oral administration, with a large amount of active ingredients excreted unchanged in feces. Furthermore, water-soluble macromolecules such as Poria cocos polysaccharides and lotus seed polysaccharides form a high-viscosity colloidal system in the extract. Through hydrogen bonding, they encapsulate flavonoids and saponins from jujube seed, significantly reducing the dissolution rate and final yield of the latter. Experimental data show that when Poria cocos and jujube seed are co-extracted in water, the total flavonoid dissolution rate of jujube seed is reduced by about 22% to 30% compared to single extraction. Traditional mixed extraction does not consider the physicochemical interactions between components, resulting in the hidden loss of effective components. More significantly, spinosin in jujube seed—a heat-sensitive component—is easily decomposed and inactivated by prolonged heating above 90 degrees Celsius, while iridoid glycosides such as monoglobulin and loganin in Cornus officinalis require higher temperatures to fully dissolve. Traditional extraction treats these components equally without establishing a temperature-zoned or staged extraction strategy, resulting in the simultaneous destruction of heat-sensitive components and insufficient extraction of heat-resistant components.

[0005] The lag in dosage form design further exacerbates the disconnect between efficacy and user experience. Human sleep has a distinct temporal structure: the sleep onset phase requires a rapid reduction in cortical excitability to shorten latency; the deep sleep phase requires maintaining stable blood drug concentrations to prolong slow-wave sleep duration; and the latter half of the night requires avoiding awakenings caused by a sudden drop in blood drug concentration. Existing oral solutions and granules require gastrointestinal dissolution, absorption, and distribution after oral administration, with peak blood drug concentration typically reaching its peak at 1.5 to 2.5 hours. The sleep onset latency is not significantly shortened, and the short half-life means that blood drug concentrations drop below the effective threshold in the latter half of the night, making patients prone to awakening and early awakening. Ordinary tablets and capsules exhibit significant differences in disintegration time, with some products not completely disintegrating within 30 minutes, resulting in delayed onset of action. They also lack sustained-release structures, failing to maintain blood drug concentrations throughout the night. Current dosage forms are not designed with release kinetics tailored to the sleep phases, and the contradiction between slow onset and short duration of action remains unresolved.

[0006] Crucially, the regulatory mechanisms of the gut-brain axis have not yet been incorporated into the core considerations of formulation design using current technologies. Modern research has confirmed a bidirectional causal relationship between gut microbiota dysbiosis and insomnia. Insomnia leads to a decrease in gut microbiota diversity and the proliferation of pathogenic bacteria, while dysbiosis affects neurotransmitter synthesis through the gut-brain axis. Gut microbiota metabolize dietary fiber to produce short-chain fatty acids, which can regulate brain function by activating the vagus nerve or directly entering the circulatory system. Some strains, such as Bifidobacterium and Lactobacillus, can synthesize neurotransmitter precursors such as γ-aminobutyric acid (GABA) and serotonin (SHT). Dysbiosis can also lead to lipopolysaccharide (LPS) entering the bloodstream, triggering systemic low-grade inflammation, increasing pro-inflammatory factors such as interleukin-6 and tumor necrosis factor-α, and disrupting the blood-brain barrier function. However, existing TCM-based calming compositions rarely incorporate formulations and processes designed around the gut-brain axis. At the formulation level, prebiotics are not specifically introduced to promote beneficial bacteria growth, nor is probiotic fermentation utilized to achieve biotransformation of active ingredients. At the process level, an integrated strategy for fermentation pretreatment, stepwise extraction, and dosage form control has not been established, making it impossible to simultaneously achieve gut microbiota regulation and component delivery. Existing technologies treat component extraction and gut microbiota regulation separately, breaking the link between formulation, gut microbiota, and brain function, thus failing to leverage the natural advantages of food-medicine homologous substances in gut-brain axis regulation.

[0007] In summary, existing technologies have significant shortcomings in terms of raw material safety, scientific extraction process, dosage form compatibility, and gut-brain axis integrated design. There is an urgent need for a sleep-aid composition and its preparation method that is based on food and medicine homologous substances, integrates biomimetic fermentation biotransformation and staged extraction technology, and constructs a rapid-release and sustained-release dual-phase release system targeting the sleep phase. Summary of the Invention

[0008] Purpose of the invention

[0009] This invention provides a sleep aid composition made entirely of substances that are both medicinal and edible. Through biomimetic fermentation pretreatment and phase gradient extraction processes, the bioavailability of active ingredients is improved, achieving the dual effects of rapid sleep onset and sleep maintenance.

[0010] To address the shortcomings of existing technologies, this invention provides a biomimetic fermentation-based sleep aid composition derived from both food and medicine, and its preparation method.

[0011] To achieve the above objectives, the present invention adopts the following technical solution: a biomimetic fermentation-based sleep-aiding food-medicine composition, comprising the following components by weight:

[0012] 15-30 portions of jujube seed;

[0013] 10-15 servings of longan pulp;

[0014] 10-15 servings of lilies;

[0015] 10-20 parts of Poria cocos;

[0016] 10-15 portions of black sesame seeds;

[0017] 10-15 servings of lotus seeds;

[0018] 10-15 portions of jujubes;

[0019] Licorice 3-6 parts;

[0020] 15-30 servings of oysters;

[0021] Cornus officinalis 6-12 parts;

[0022] 5-10 portions of peach kernels;

[0023] 6-10 parts of Lophatherum gracile.

[0024] The preferred ingredients are: 20-28 parts of jujube seed; 11-14 parts of longan pulp; 11-14 parts of lily bulb; 12-18 parts of poria cocos; 11-14 parts of black sesame; 11-14 parts of lotus seed; 11-14 parts of jujube; 4-5 parts of licorice root; 20-28 parts of oyster shell; 7-10 parts of cornus officinalis; 6-8 parts of peach kernel; and 7-9 parts of lophatherum gracile.

[0025] The formulation compatibility mechanism of the above technical solution is as follows:

[0026] The core ingredients for calming the mind and soothing the nerves are: jujube seed, lily bulb, lotus seed, and longan pulp—which nourish the heart and spleen and replenish the heart blood. Jujube seed saponins directly regulate the central GABAergic system, while lily bulb and lotus seed work together to calm the nerves, and longan pulp nourishes the heart and spleen to soothe the mind and soothe the nerves.

[0027] The sedative and calming group: Oyster - It has the functions of calming the mind and relieving anxiety, and is used for insomnia caused by yin deficiency and yang hyperactivity.

[0028] Tonifying and astringent group: Cornus officinalis – tonifies the liver and kidneys, astringes and consolidates, and helps to promote sleep by consolidating essence and qi.

[0029] Spleen-strengthening and Qi-boosting, gut-brain axis regulation group: Licorice, jujube, Poria cocos - strengthens the spleen and harmonizes the middle jiao, Poria cocos polysaccharide, as a prebiotic, indirectly regulates sleep through the gut-brain axis, and licorice harmonizes the other herbs;

[0030] Blood-activating and meridian-clearing group: Peach kernel - activates blood circulation and removes blood stasis, improves cerebral microcirculation, and promotes the delivery of calming components;

[0031] The "Calming the Mind and Relieving Irritability" group: Lophatherum gracile – clears the mind and promotes urination, eliminates neuroinflammatory factors, and improves cerebral microcirculation;

[0032] Liver and kidney tonic, brain nourishing group: Black sesame seeds - nourish the liver and kidneys, replenish essence and blood, rich in unsaturated fatty acids, vitamin E and magnesium, nourish nerves and resist oxidative stress.

[0033] Preferably, the composition comprises an immediate-release component and a sustained-release component, wherein the weight ratio of the immediate-release component to the sustained-release component is 1:1 to 2:1; the immediate-release component comprises fermentation supernatant obtained from jujube kernel, peach kernel and black sesame seeds after biomimetic fermentation treatment and alcohol extract concentrate; the sustained-release component comprises an aqueous extract.

[0034] The oysters are added to the composition after being cleaned, dried, and ultra-finely pulverized.

[0035] This invention also provides a method for preparing a biomimetic fermented sleep-aiding food-medicine homology composition, comprising the following steps:

[0036] S1. Bionic fermentation pretreatment;

[0037] S2, alcohol extraction;

[0038] S3, water extraction;

[0039] S4. Preparation of immediate-release components;

[0040] S5. Preparation of sustained-release components;

[0041] S6, Mixed molding.

[0042] Preferably, step S1 includes: grinding jujube kernels, peach kernels and black sesame seeds into 40-80 mesh and then mixing them;

[0043] Add 8 to 12 times its weight of purified water;

[0044] Sterilize at 115–121°C for 15–30 min (before sterilization, add 0.01%–0.05% by weight of natural antioxidants, such as vitamin E, tea polyphenols, or rosemary extract, to the fermentation system to inhibit the oxidation of unsaturated fatty acids during high-temperature sterilization).

[0045] After cooling to 35-38℃, inoculate with compound probiotics, purge the air in the fermenter with nitrogen until the oxygen content is ≤0.5%, maintain a positive pressure of 0.02-0.05MPa in the tank, and ferment under static anaerobic conditions for 18-24 hours;

[0046] After fermentation, solid-liquid separation was performed to obtain fermentation supernatant A and fermentation residue D;

[0047] Preferably, the compound probiotics include Lactobacillus plantarum, Lactobacillus acidophilus, and Bifidobacterium longum;

[0048] The mass ratio of *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium longum* is (0.5–2):(0.5–2):(0.5–2).

[0049] The amount of compound probiotics inoculated is 2-5% of the total weight of the fermentation system.

[0050] By employing the aforementioned technologies, this step simulates the human intestinal environment. The glycosidases produced by probiotics can convert jujube seed saponin A into aglycones, gradually degrade amygdalin in peach kernels into active small molecules, and hydrolyze black sesame protein into absorbable small peptides containing tryptophan, thereby significantly improving the bioavailability of the three types of fat-soluble / macromolecule components.

[0051] Preferably, S2 includes:

[0052] Fermentation residue D was mixed with lily bulb, lotus seed, poria cocos and light bamboo leaf (light bamboo leaf is an extract targeting flavonoid components);

[0053] Add 6 to 10 times its weight of an ethanol solution with a volume fraction of 60 to 80%;

[0054] Reflux extraction at 70–90℃ 2–3 times;

[0055] Each extraction takes 1–3 hours;

[0056] The extracts were combined after filtration;

[0057] The alcohol concentrate B was obtained by vacuum concentration.

[0058] Preferably, S3 includes:

[0059] The residue after alcohol extraction was mixed with longan pulp, jujube, licorice, and Cornus officinalis (the iridoid glycosides in Cornus officinalis);

[0060] Add 8 to 12 times the weight of purified water;

[0061] Extract 2-3 times at 90-100℃;

[0062] Each extraction takes 1-2 hours;

[0063] The extracts were combined after filtration;

[0064] Reduced pressure concentration yields water extract C.

[0065] Preferably, S4 includes:

[0066] Mix fermentation supernatant A with alcohol extract concentrate B;

[0067] After adding excipients, spray drying or freeze drying is performed;

[0068] The immediate-release component E was obtained;

[0069] The excipients include one or more of maltodextrin, resistant dextrin, and microcrystalline cellulose.

[0070] Using the above technical solution, the S1 biomimetic fermentation pretreatment is carried out at a low temperature of 35~38℃. The heat-sensitive component of spinosin in jujube seed is mainly concentrated in the fermentation supernatant A, which is directly used in the preparation of the S4 immediate-release component without high-temperature treatment. The S2 alcohol extraction targets fermentation residue D and lily, lotus seed, poria cocos, and lophatherum gracile. Among them, the target components such as lily saponins, lotus seed alkaloids, poria cocos triterpenes, and lophatherum gracile flavonoids have good heat resistance, and alcohol extraction at 70~90℃ can effectively dissolve them without significant degradation.

[0071] The raw material for S3 water extraction is the residue after alcohol extraction, in which the heat-sensitive component (spinoxanol) has been enriched in the supernatant A in S1. The target components for S3 water extraction are cornus iridoid glycosides (monoside, loganin), glycyrrhizic acid, jujube polysaccharides, etc. The above components have good stability under water extraction conditions of 90~100℃.

[0072] Preferably, S5 and S6 include:

[0073] The concentrated aqueous extract C was mixed with the sustained-release carrier and then granulated to obtain the sustained-release component F.

[0074] The sustained-release carrier is one or more of hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and pectin;

[0075] The immediate-release component E and the sustained-release component F are mixed in a certain proportion, and then oyster ultrafine powder is added (the main active ingredients of oyster are calcium carbonate, amino acids and trace elements, which are easily reduced in utilization by alcohol extraction or water extraction, so ultrafine powder is added directly). After mixing evenly, tablets, capsules, granules, solid beverages, compressed candies or powders are prepared to obtain the biomimetic fermented sleep aid food and medicine homology composition.

[0076] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0077] 1) This invention uses a sleep aid composition made entirely of food and medicine homologous substances. Through the synergistic combination of raw materials such as jujube seed, longan pulp, lily bulb, lotus seed, poria cocos, and black sesame, it achieves multiple regulation pathways such as nourishing the heart and calming the mind, strengthening the spleen and replenishing qi, tonifying the liver and kidneys, and clearing the mind and relieving irritability. It has high safety and is suitable for long-term consumption.

[0078] 2) This invention employs a biomimetic fermentation pretreatment process, utilizing compound probiotics to biotransform some bound active ingredients in jujube kernels, peach kernels, and black sesame seeds, transforming them into smaller molecule active substances that are more easily absorbed and utilized, thereby improving the utilization efficiency of the active ingredients in the raw materials.

[0079] 3) This invention employs a phase-separated gradient extraction process that combines alcohol extraction and water extraction to achieve the classification and enrichment of active ingredients with different polarities, avoiding the problem of mutual interference of active ingredients in the traditional mixed extraction process, and improving the comprehensive utilization rate of active ingredients in the raw materials.

[0080] 4) This invention constructs a biphasic release system that combines immediate-release and sustained-release components, and directly adds oysters in the form of ultrafine powder to the product to achieve synergistic utilization of active ingredients and mineral components, thereby improving the stability and application value of the product. Attached Figure Description

[0081] Figure 1 This is a diagram illustrating the biomimetic fermentation mechanism.

[0082] Figure 2 This is a diagram of a two-phase release structure;

[0083] Figure 3 Diagram showing the sleep latency period;

[0084] Figure 4 A graph showing sleep duration;

[0085] Figure 5 To release the curve;

[0086] Figure 6 This is a group diagram of the gut-brain axis. Detailed Implementation

[0087] The present invention will be further described below through specific embodiments. To make the inventive objectives, technical solutions, and beneficial technical effects of the present invention clearer, the present invention will be further described in detail below with reference to the embodiments. It should be understood that the embodiments described in this specification are merely for explaining the present invention and are not intended to limit the present invention.

[0088] Unless otherwise stated, all instruments and reagents used in the examples are commercially available or synthesized using conventional methods and can be used directly without further processing, and all instruments used in the examples are commercially available.

[0089] Example 1:

[0090] 1. The formula, by weight, includes the following components:

[0091] 15 parts of jujube seed, 10 parts of longan pulp, 10 parts of lily bulb, 10 parts of poria cocos, 10 parts of black sesame, 10 parts of lotus seed, 10 parts of jujube, 3 parts of licorice root, 15 parts of oyster, 6 parts of cornus officinalis, 5 parts of peach kernel, and 6 parts of bamboo leaf.

[0092] 2. Preparation method

[0093] S1, Bionic Fermentation Pretreatment

[0094] Sour jujube kernels, peach kernels, and black sesame seeds were separately pulverized to 40 mesh, then mixed together. Eight times their weight of purified water were added, and the mixture was sterilized at 115℃ for 30 minutes. After cooling to 35℃, a compound probiotic was inoculated. The compound probiotic included *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium longum* in a mass ratio of 0.5:0.5:0.5, with an inoculation amount of 2% of the total weight of the fermentation system. Fermentation was carried out under anaerobic conditions for 18 hours. After fermentation, solid-liquid separation was performed to obtain fermentation supernatant A and fermentation residue D.

[0095] S2, alcohol extraction

[0096] Fermentation residue D was mixed with lily bulb, lotus seed, poria cocos and bamboo leaf, and 6 times its weight of 60% ethanol solution was added. The mixture was refluxed twice at 70°C for 1 hour each time. The extracts were combined after filtration and concentrated under reduced pressure to obtain concentrated ethanol extract B.

[0097] S3, Water Extraction

[0098] The residue after alcohol extraction was mixed with longan pulp, jujube, licorice and cornus officinalis, and 8 times the weight of purified water was added. The mixture was extracted twice at 90℃ for 1 hour each time. After filtration, the extracts were combined and concentrated under reduced pressure to obtain concentrated water extract C.

[0099] S4, Preparation of immediate-release components

[0100] Fermentation supernatant A was mixed with alcohol extract concentrate B, maltodextrin was added as an excipient, and spray-dried to obtain immediate-release component E.

[0101] S5. Preparation of sustained-release components

[0102] The concentrated aqueous extract C was mixed with hydroxypropyl methylcellulose and then granulated to obtain the sustained-release component F.

[0103] S6, Mixed Molding

[0104] Immediate-release component E and sustained-release component F are mixed in a weight ratio of 1:1, and then oyster ultrafine powder is added and mixed evenly to form tablets, thus obtaining the biomimetic fermented sleep aid food-medicine homology composition.

[0105] 3. Quality Inspection

[0106] The resulting tablets are brownish-yellow in color, with a slightly fragrant odor and a slightly sweet taste. According to the methods specified in the Chinese Pharmacopoeia, the content of jujube seed saponin A is 3.2 mg / g, and the total flavonoid content is 12.5 mg / g.

[0107] 4. Effect Verification

[0108] 4.1 Mouse pentobarbital-induced sleep synergy experiment

[0109] Sixty ICR mice were randomly divided into six groups (blank control group, positive control group (diazepam tablets (2 mg / kg) + jujube seed extract monomer group), low-dose group of Example 1, medium-dose group, high-dose group, and comparative example 1 group), with 10 mice in each group, half male and half female. The mice were administered the drug by gavage for 7 consecutive days. Thirty minutes after the last administration, each group was injected intraperitoneally with sodium pentobarbital 45 mg / kg. The disappearance of the righting reflex was used as the indicator of sleep onset, and the sleep latency and duration of sleep were recorded.

[0110] Results: In Example 1, the sleep latency of the medium dose group (equivalent to 10 times the recommended human dose) was 14.8±2.3 min and the sleep duration was 89.5±7.2 min. Compared with the blank control group (latency 22.6±3.1 min, duration 58.3±5.7 min), the difference was statistically significant (P<0.01).

[0111] 4.2 In vitro release rate determination

[0112] The release rate was determined according to the method specified in the Chinese Pharmacopoeia (slurry method, rotation speed 75 r / min, medium: 900 mL pH 6.8 phosphate buffer, temperature 37℃). Results: The release rate at 30 min was 42.6% (basic release of the immediate release fraction), the cumulative release rate at 6 h was 68.3%, and the cumulative release rate at 12 h was 85.2%.

[0113] 4.3 Gut-brain axis index detection

[0114] Fourteen days after drug administration, mouse feces were collected, and the content of short-chain fatty acids was determined by GC-MS. Results: Acetic acid content was 38.4±3.2 μmol / g, propionic acid content was 15.6±1.8 μmol / g, and butyric acid content was 8.3±0.9 μmol / g, representing increases of 32.5%, 28.7%, and 35.2%, respectively, compared to the blank control group (P<0.05). 16S rRNA sequencing showed that the Shannon index increased from 3.82±0.21 in the blank control group to 4.45±0.18 (P<0.05).

[0115] Example 2:

[0116] 1. The formula, by weight, includes the following components: 24 parts of jujube seed, 12 parts of longan pulp, 12 parts of lily bulb, 15 parts of poria cocos, 12 parts of black sesame, 12 parts of lotus seed, 12 parts of jujube, 4.5 parts of licorice root, 24 parts of oyster shell, 8 parts of cornus officinalis, 7 parts of peach kernel, and 8 parts of bamboo leaf.

[0117] 2. Preparation method

[0118] S1, Bionic Fermentation Pretreatment

[0119] Sour jujube kernels, peach kernels, and black sesame seeds were separately pulverized to 60 mesh and then mixed. Ten times their weight of purified water were added, and the mixture was sterilized at 121℃ for 20 minutes. After cooling to 37℃, a compound probiotic was inoculated. The compound probiotic included *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium longum* in a mass ratio of 1:1:1, and the inoculation amount was 3.5% of the total weight of the fermentation system. Fermentation was carried out under anaerobic conditions for 21 hours. After fermentation, solid-liquid separation was performed to obtain fermentation supernatant A and fermentation residue D.

[0120] S2, alcohol extraction

[0121] Fermentation residue D was mixed with lily bulb, lotus seed, poria cocos and bamboo leaf, and 8 times its weight of 70% ethanol solution was added. The mixture was refluxed twice at 80℃ for 2 hours each time. After filtration, the extracts were combined and concentrated under reduced pressure to obtain concentrated ethanol extract B.

[0122] S3, Water Extraction

[0123] The residue after alcohol extraction was mixed with longan pulp, jujube, licorice and cornus officinalis, and 10 times the weight of purified water was added. The mixture was extracted twice at 95°C for 1.5 hours each time. After filtration, the extracts were combined and concentrated under reduced pressure to obtain concentrated water extract C.

[0124] S4, Preparation of immediate-release components

[0125] Fermentation supernatant A was mixed with alcohol extract concentrate B, resistant dextrin was added as an excipient, and the mixture was freeze-dried to obtain immediate-release component E.

[0126] S5. Preparation of sustained-release components

[0127] The concentrated aqueous extract C was mixed with sodium alginate and then granulated to obtain the sustained-release component F.

[0128] S6, Mixed Molding

[0129] Immediate-release component E and sustained-release component F are mixed at a weight ratio of 1.5:1, and then oyster ultrafine powder is added and mixed evenly to form capsules, thus obtaining the biomimetic fermented sleep aid food-medicine homology composition.

[0130] 3. Quality Inspection

[0131] The contents of the resulting capsules were a brownish-yellow powder with a slightly fragrant odor and a slightly sweet taste. The content of jujube seed saponin A was 4.8 mg / g, and the total flavonoid content was 16.2 mg / g.

[0132] 4. Effect Verification

[0133] 4.1 Mouse pentobarbital-induced sleep synergy experiment

[0134] The grouping and administration methods were the same as in Example 1. Results: In Example 2, the sleep latency in the medium-dose group was 11.2±1.8 min, and the sleep duration was 102.6±8.5 min, which were significantly different from the blank control group (P<0.01) and better than the same-dose group in Example 1 (P<0.05).

[0135] 4.2 In vitro release rate determination

[0136] The measurement conditions were the same as in Example 1. Results: The release rate at 30 min was 48.3% (the immediate-release portion was basically released), the cumulative release rate at 6 h was 75.6%, and the cumulative release rate at 12 h was 91.8%. The sustained-release effect was better than that in Example 1.

[0137] 4.3 Gut-brain axis index detection

[0138] Mouse feces were collected 14 days after drug administration. Results: Acetic acid content was 45.2±3.8 μmol / g, propionic acid content was 18.3±2.1 μmol / g, and butyric acid content was 9.7±1.1 μmol / g, which were increased by 55.8%, 51.2%, and 58.3% respectively compared with the blank control group (P<0.01). The Shannon index increased to 4.72±0.15 (P<0.01).

[0139] 4.4 Human Trial Testing

[0140] A randomized, double-blind, controlled study was conducted, recruiting 120 volunteers with mild insomnia. They were randomly assigned to three groups: Example 2 group, a positive control group (melatonin tablets), and a placebo group, with 40 participants in each group. All participants received melatonin for 28 consecutive days. The primary endpoints were the Pittsburgh Sleep Quality Index (PSQI) total score and sleep efficiency (sleep efficiency = actual sleep time / time spent in bed × 100%).

[0141] Results: In Example 2, the total PSQI score decreased from baseline 14.2±2.6 to 6.8±1.9 (P<0.01), and sleep efficiency increased from 68.5±8.3% to 85.2±6.7% (P<0.01). Compared with the positive control group, the degree of improvement in PSQI was comparable (P>0.05), but the improvement in sleep efficiency was more significant (P<0.05), and no adverse reactions such as morning dizziness or daytime sleepiness occurred.

[0142] Example 3:

[0143] 1. Formula: Take the weighted values ​​at both ends, 28 parts of jujube seed, 14 parts of longan pulp, 14 parts of lily bulb, 18 parts of poria cocos, 14 parts of black sesame, 14 parts of lotus seed, 14 parts of jujube, 5 parts of licorice, 28 parts of oyster, 10 parts of cornus officinalis, 8 parts of peach kernel, and 9 parts of bamboo leaf.

[0144] 2. Preparation method: S1 is pulverized to 80 mesh, added with 12 times the amount of water, sterilized at 121℃ for 15 min, inoculated at 5%, with a bacterial culture ratio of 2:2:2, and fermented for 24 h; S2 is extracted with 10 times the amount of 70% ethanol at 90℃ three times, each time for 3 h; S3 is extracted with 12 times the amount of water at 100℃ three times, each time for 2 h; S4 is excipient microcrystalline cellulose, spray dried; S5 is a slow-release carrier of pectin; S6 is a rapid release:slow release ratio of 2:1, and granules are prepared.

[0145] 3. Effects: Sleep latency 10.5±1.6 min, sleep duration 106.3±9.1 min, and release rate 93.5% at 12 hours.

[0146] Example 4:

[0147] 1. Formula: Same as Example 2.

[0148] 2. Preparation method: Same as in Example 2, except that in S6, the immediate-release component E and the sustained-release component F are mixed in a weight ratio of 1:1.

[0149] 3. Results: Sleep latency was 11.8 ± 2.0 min, sleep duration was 98.4 ± 7.8 min, release rate was 72.3% at 6 hours and 88.5% at 12 hours. The immediate release effect was slightly faster, while the sustained release duration was slightly shorter than in Example 2.

[0150] Example 5:

[0151] 1. Formula: Same as Example 2.

[0152] 2. Preparation method: Same as in Example 2, except that in S6, the immediate-release component E and the sustained-release component F are mixed at a weight ratio of 2:1.

[0153] 3. Effects: Sleep latency was 10.8±1.7 min, sleep duration was 99.2±8.1 min, release rate was 78.2% at 6 hours and 90.1% at 12 hours. The proportion of immediate release increased and the sleep latency was further shortened, but the sustained-release capacity decreased slightly.

[0154] Comparative Example 1:

[0155] Traditional Jujube Seed Soup Recipe: 15 parts jujube seed, 10 parts anemarrhena rhizome, 10 parts poria cocos, 6 parts chuanxiong rhizome, and 3 parts licorice root.

[0156] The preparation method involves mixing all raw materials, adding 10 times the amount of water, decocting twice for 1 hour each time, combining the filtrates, concentrating and drying.

[0157] Results: Sleep latency was 17.6±2.8 min, and sleep duration was 71.3±6.5 min. The improvement in PSQI was weaker than in Example 2 (P<0.05). During the trial, 3 cases experienced stomach discomfort, and 2 cases experienced dry mouth.

[0158] This demonstrates that non-food and medicine homologous herbs (Anemarrhena asphodeloides and Ligusticum chuanxiong) have a narrow safety window and are not suitable for daily health maintenance.

[0159] Comparative Example 2:

[0160] The same formulation and preparation process as in Example 2, except that probiotic inoculation and fermentation are not performed in S1.

[0161] Preparation method: Grind jujube kernel, peach kernel and black sesame seeds into 60 mesh and mix them. Add 10 times the weight of purified water, sterilize at 121℃ for 20 min, cool to 37℃ and let stand at the same temperature for 21 h without inoculation. Then perform solid-liquid separation to obtain unfermented supernatant A′ and unfermented residue D′. Unfermented supernatant A′ enters S4 and is mixed with alcohol extract concentrate B. Unfermented residue D′ enters S2 for alcohol extraction. The remaining steps S2 to S6 are the same as in Example 2.

[0162] Effects: The conversion rate of jujube seed saponin A aglycone was only 9.8%, a decrease of 88.1% compared to 82.6% in Example 2 (P<0.01); the area under the blood drug concentration-time curve (AUC) was also lower. 0-12h Compared with Example 2, the sleep latency was reduced by 71.6% (P<0.01); the sleep latency was 15.2±2.4 min and the sleep duration was 81.3±6.8 min, both of which were worse than Example 2 (P<0.01).

[0163] The study demonstrated that, under the same physical treatment conditions, the elimination of biomimetic fermentation resulted in the loss of saponin biotransformation and a significant decrease in bioavailability, thus confirming the crucial role of the fermentation step.

[0164] Comparative Example 3:

[0165] Same formulation as Example 2, but using "one-pot" alcohol extraction.

[0166] The preparation method involves mixing all raw materials (except oysters), adding 8 times the amount of 70% ethanol, reflux extraction twice, 2 hours each time, combining the filtrates, concentrating and drying them, and then mixing them with oyster ultrafine powder.

[0167] Results: The polysaccharide yield decreased by 68.5% compared with Example 2 (P<0.01); the short-chain fatty acid content was not significantly different from the blank control group (P>0.05); the sleep latency was 13.8±2.2 min, and the sleep duration was 82.4±7.3 min.

[0168] This demonstrates that alcohol extraction cannot extract water-soluble polysaccharides, resulting in the loss of gut-brain axis regulation.

[0169] Comparative Example 4:

[0170] The formula is the same as in Example 2, but the step-by-step extraction is cancelled, and the "one-pot cooking after full-formula fermentation" process is adopted.

[0171] Preparation method: Grind jujube kernel, peach kernel and black sesame seeds into 60 mesh and mix them. Add 10 times the weight of purified water and sterilize at 121℃ for 20 min. After cooling to 37℃, inoculate with compound probiotics (Lactobacillus plantarum, Lactobacillus acidophilus and Bifidobacterium longum in a mass ratio of 1:1:1 and an inoculation amount of 3.5%) and anaerobic ferment for 21 h. After fermentation, mix all fermentation products (supernatant + residue) with lily bulb, lotus seed, poria cocos, light bamboo leaf, longan pulp, jujube, licorice and cornus officinalis, add 10 times the amount of water, and decoct twice at 95℃ for 1.5 h each time. Filter, concentrate and dry the filtrate and mix it with oyster ultrafine powder to granulate.

[0172] Results: The total flavonoid yield of jujube seed decreased by 24.6% compared with Example 2 (P<0.01), and the monoglycoside yield decreased by 18.3% (P<0.05); the active aglycones produced by fermentation were further decomposed and lost by 37.2% during high-temperature decoction (P<0.01); the sleep latency was 13.8±2.2 min, and the sleep duration was 84.5±7.1 min, both of which were worse than those in Example 2 (P<0.01).

[0173] The results show that direct high-temperature cooking without step-by-step extraction after fermentation leads to the simultaneous destruction of fermentation products and antagonistic dissolution of components, thus confirming the necessity of the integrated design of "fermentation pretreatment + step-by-step extraction".

[0174] Comparative Example 5:

[0175] Same as Example 2, but the compound probiotics are only inoculated with Lactobacillus plantarum.

[0176] The preparation method involves inoculating only *Lactobacillus plantarum* in S1 at an inoculation rate of 3.5% and fermenting for 21 hours.

[0177] Results: The conversion rate of jujube seed saponin A aglycone was 58.3%, which was lower than 82.6% in Example 2 (P<0.01); the total amount of short-chain fatty acids decreased by 31.2% compared with Example 2 (P<0.05); the sleep latency was 13.2±2.1 min, and the sleep duration was 88.5±7.6 min.

[0178] This demonstrates that the fermentation effect of a single strain is not as good as that of a synergistic effect of a compound strain.

[0179] Comparative Example 6:

[0180] Same as Example 2, but without the distinction between immediate / slow-release; all are mixed and granulated.

[0181] The preparation method involves combining fermentation supernatant A, alcohol extract concentrate B, and water extract concentrate C, adding excipients for granulation, and then adding oyster ultrafine powder for mixing and tableting.

[0182] Results: The release rate at 30 min was only 18.5% (48.3% in Example 2), and the sleep latency was prolonged to 15.6 ± 2.4 min (P < 0.01); the cumulative release rate at 6 h reached 92.7% (75.6% in Example 2), and there was no significant increase in the cumulative release rate at 12 h. The number of awakenings in the second half of the night increased by 2.3 times compared to Example 2 (P < 0.05).

[0183] The lack of an immediate / slow-release design resulted in a slow onset and short duration of action, failing to match sleep phases.

[0184] Comparative Example 7:

[0185] Same as Example 2, but oysters are added in S3 water extraction instead of in the form of ultrafine powder.

[0186] The preparation method involves water extraction of oysters together with longan pulp, jujube, licorice, and cornus officinalis, without separate ultrafine pulverization.

[0187] Effects: The calcium ion dissolution rate was improved, but the sedative active ingredients (such as oyster polypeptides and amino acids) were destroyed by high temperature extraction, with a loss rate of 45.6% (P<0.01); the sleep latency was 12.8±2.0 min and the sleep duration was 93.4±7.8 min, both of which were worse than those in Example 2 (P<0.05).

[0188] This study demonstrates that adding ultrafine oyster powder directly can retain its heat-sensitive sedative components.

[0189] Comparative Example 8:

[0190] Same as Example 2, but remove Poria cocos and replace it with an equal amount of starch.

[0191] The preparation method is the same as in Example 2.

[0192] Results: The total amount of short-chain fatty acids decreased by 42.6% compared with Example 2 (P<0.01), and the Shannon index did not change significantly (P>0.05); the sleep latency was 13.5±2.3 min and the sleep duration was 86.2±7.1 min, both of which were worse than those in Example 2 (P<0.05).

[0193] This demonstrates the necessity of Poria cocos as a prebiotic for regulating the gut-brain axis.

[0194] Comparative Example 9:

[0195] Comparison of commercially available jujube seed oil soft capsules (Yiling brand jujube seed oil soft capsules, National Food and Drug Administration Registration No. G20160175).

[0196] Preparation method: Take the product according to the recommended dosage in the product instructions, and carry out the same procedure and conditions as in Example 2;

[0197] Results: The PSQI total score decreased from baseline 14.5±2.4 to 9.2±2.1 (P<0.01), a weaker improvement than in Example 2 (6.8±1.9, P<0.05); sleep efficiency increased from 67.8±8.1% to 76.5±7.2% (P<0.01), lower than in Example 2 (85.2±6.7%, P<0.05). Five cases experienced mild diarrhea.

[0198] This demonstrates that commercially available single-extract products are less effective than the compound formula of this invention, and lack gut-brain axis regulation.

[0199] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A biomimetic fermentation-based sleep-aiding food-medicine composition, characterized by, by weight parts, It includes the following components: 15-30 portions of jujube seed; 10-15 servings of longan pulp; 10-15 servings of lilies; 10-20 parts of Poria cocos; 10-15 portions of black sesame seeds; 10-15 servings of lotus seeds; 10-15 portions of jujubes; Licorice 3-6 parts; 15-30 servings of oysters; Cornus officinalis 6-12 parts; 5-10 portions of peach kernels; 6-10 parts of Lophatherum gracile.

2. The biomimetic fermented sleep-aiding food-medicine homology composition according to claim 1, characterized in that, Sour jujube seed 20-28 parts; longan pulp 11-14 parts; lily bulb 11-14 parts; poria cocos 12-18 parts; black sesame 11-14 parts; lotus seed 11-14 parts; jujube 11-14 parts; licorice root 4-5 parts; oyster shell 20-28 parts; cornus officinalis 7-10 parts; peach kernel 6-8 parts; lophatherum gracile 7-9 parts.

3. The biomimetic fermentation-based sleep-aiding food-medicine homology composition according to claim 1, characterized in that, The composition comprises an immediate-release component and a sustained-release component, wherein the weight ratio of the immediate-release component to the sustained-release component is 1:1 to 2:1; the immediate-release component comprises fermentation supernatant obtained from jujube kernel, peach kernel and black sesame seeds after biomimetic fermentation treatment and alcohol extract concentrate; the sustained-release component comprises an aqueous extract. The oysters are added to the composition after being cleaned, dried, and ultra-finely pulverized.

4. A method for preparing the composition according to any one of claims 1 to 3, characterized in that, Includes the following steps: S1. Bionic fermentation pretreatment; S2, alcohol extraction; S3, water extraction; S4. Preparation of immediate-release components; S5. Preparation of sustained-release components; S6, Mixed molding.

5. The preparation method of the biomimetic fermentation-based sleep-aiding food-medicine homology composition according to claim 4, characterized in that, S1 includes: Grind the jujube kernels, peach kernels, and black sesame seeds separately to 40-80 mesh and then mix them together. Add 8 to 12 times its weight of purified water; Sterilize at 115–121℃ for 15–30 minutes; After cooling to 35–38°C, inoculate with compound probiotics; Fermentation under anaerobic conditions for 18–24 hours; After fermentation, solid-liquid separation was performed to obtain fermentation supernatant A and fermentation residue D.

6. The preparation method of the biomimetic fermentation-based sleep-aiding food-medicine homology composition according to claim 5, characterized in that, The compound probiotics include Lactobacillus plantarum, Lactobacillus acidophilus, and Bifidobacterium longum. The mass ratio of *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium longum* is (0.5–2):(0.5–2):(0.5–2). The amount of compound probiotics inoculated is 2-5% of the total weight of the fermentation system.

7. The preparation method of the biomimetic fermentation-based sleep-aiding food-medicine homology composition according to claim 4, characterized in that, S2 includes: Mix fermentation residue D with lily bulbs, lotus seeds, poria cocos, and bamboo leaves; Add 6 to 10 times its weight of an ethanol solution with a volume fraction of 60 to 80%; Reflux extraction at 70–90℃ 2–3 times; Each extraction takes 1-3 hours; The extracts were combined after filtration; The concentrated alcohol extract B was obtained by vacuum concentration.

8. The preparation method of the biomimetic fermentation-based sleep-aiding food-medicine homology composition according to claim 4, characterized in that, S3 includes: The residue after alcohol extraction is mixed with longan pulp, jujube, licorice and cornus officinalis; Add 8 to 12 times the weight of purified water; Extract 2-3 times at 90-100℃; Each extraction takes 1-2 hours; The extracts were combined after filtration; Reduced pressure concentration yields water extract C.

9. The preparation method of the biomimetic fermented sleep-aiding food-medicine homology composition according to claim 4, characterized in that, S4 includes: Mix fermentation supernatant A with alcohol extract concentrate B; After adding excipients, spray drying or freeze drying is performed; The immediate-release component E was obtained; The excipients include one or more of maltodextrin, resistant dextrin, and microcrystalline cellulose.

10. The preparation method of a biomimetic fermented sleep-aiding food-medicine homology composition according to claim 4, characterized in that, S5 and S6 include: The concentrated aqueous extract C was mixed with the sustained-release carrier and then granulated to obtain the sustained-release component F. The sustained-release carrier is one or more of hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and pectin; The immediate-release component E and the sustained-release component F are mixed in a certain proportion, and then oyster ultrafine powder is added and mixed evenly to make tablets, capsules, granules, solid beverages, compressed candies or powders, thus obtaining the biomimetic fermented sleep aid food and medicine homology composition.

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